Yan Liu

The importance of the p53 protein in the cellular response to DNA damage is well known, but its function during steady-state hematopoiesis has not been established. We have defined a critical role of p53 in regulating hematopoietic stem cell quiescence, especially in promoting the enhanced quiescence seen in HSCs that lack the MEF/ELF4 transcription factor. Transcription profiling of HSCs isolated from wild-type and p53 null mice identified Gfi-1 and Necdin as p53 target genes, and using lentiviral vectors to upregulate or knockdown the expression of these genes, we show their importance in regulating HSC quiescence. Establishing the role of p53 (and its target genes) in controlling the cell-cycle entry of HSCs may lead to therapeutic strategies capable of eliminating quiescent cancer (stem) cells.

Natural bone is a mineralized biological material, which serves a supportive and protective framework for the body, stores minerals for metabolism, and produces blood cells nourishing the body. Normally, bone has an innate capacity to heal from damage. However, massive bone defects due to traumatic injury, tumor resection, or congenital diseases pose a great challenge to reconstructive surgery. Scaffold-based tissue engineering (TE) is a promising strategy for bone regenerative medicine, because biomaterial scaffolds show advanced mechanical properties and a good degradation profile, as well as the feasibility of controlled release of growth and differentiation factors or immobilizing them on the material surface. Additionally, the defined structure of biomaterial scaffolds, as a kind of mechanical cue, can influence cell behaviors, modulate local microenvironment and control key features at the molecular and cellular levels. Recently, nano/micro-assisted regenerative medicine becomes a promising application of TE for the reconstruction of bone defects. For this reason, it is necessary for us to have in-depth knowledge of the development of novel nano/micro-based biomaterial scaffolds. Thus, we herein review the hierarchical structure of bone, and the potential application of nano/micro technologies to guide the design of novel biomaterial structures for bone repair and regeneration.

T helper 17 (TH17) cells have been shown to contribute to multiple disease systems. However, the functional phenotype and survival pattern of TH17 cells as well as the underlying mechanisms that control TH17 cells have been poorly investigated in humans, significantly hampering the clinical targeting of these cells. Here, we studied human TH17 cells in the pathological microenvironments of graft-versus-host disease, ulcerative colitis, and cancer; TH17 cell numbers were increased in the chronic phase of these diseases. Human TH17 cells phenotypically resembled terminally differentiated memory T cells but were distinct from central memory, exhausted, and senescent T cells. Despite their phenotypic markers of terminal differentiation, TH17 cells mediated and promoted long-term antitumor immunity in in vivo adoptive transfer experiments. Furthermore, TH17 cells had a high capacity for proliferative self-renewal, potent persistence, and apoptotic resistance in vivo, as well as plasticity-converting into other types of TH cells. These cells expressed a relatively specific gene signature including abundant antiapoptotic genes. We found that hypoxia-inducible factor-1α and Notch collaboratively controlled key antiapoptosis Bcl-2 family gene expression and function in TH17 cells. Together, these data indicate that human TH17 cells may be a long-lived proliferating effector memory T cell population with unique genetic and functional characteristics. Targeting TH17-associated signaling pathway would be therapeutically meaningful for treating patients with autoimmune disease and advanced tumor.

Abstract Human pulp stem cells (PSCs) include dental pulp stem cells (DPSCs) isolated from dental pulp tissues of human extracted permanent teeth and stem cells from human exfoliated deciduous teeth (SHED). Depending on their multipotency and sensitivity to local paracrine activity, DPSCs and SHED exert therapeutic applications at multiple levels beyond the scope of the stomatognathic system. This review is specifically concentrated on PSC-updated biological characteristics and their promising therapeutic applications in (pre)clinical practice. Biologically, distinguished from conventional mesenchymal stem cell markers in vitro, NG2, Gli1, and Celsr1 have been evidenced as PSC markers in vivo. Both perivascular cells and glial cells account for PSC origin. Therapeutically, endodontic regeneration is where PSCs hold the most promises, attributable of PSCs’ robust angiogenic, neurogenic, and odontogenic capabilities. More recently, the interplay between cell homing and liberated growth factors from dentin matrix has endowed a novel approach for pulp-dentin complex regeneration. In addition, PSC transplantation for extraoral tissue repair and regeneration has achieved immense progress, following their multipotential differentiation and paracrine mechanism. Accordingly, PSC banking is undergoing extensively with the intent of advancing tissue engineering, disease remodeling, and (pre)clinical treatments. Significance statement Pulp stem cells can be readily harvested from dental pulp tissue of extracted permanent teeth and exfoliated deciduous teeth, respectively. However, a systematic and comprehensive review about pulp stem cells in terms of biological attributes and therapeutic applications is lacking. Accordingly, this review is concentrated on pulp stem cells to emphasize their updated biological characteristics such as cell markers, multipotency and origin, and promising therapeutic applications, including endodontic regeneration and extraoral tissue repair and regeneration, as well as rising cell bank with the intent of enhancing the understanding of dental mesenchymal stem cells and advancing associated tissue engineering and disease treatment.

MSC-exo eye drops alleviate GVHD-associated dry eye disease by shifting M1 macrophages to M2 via miR-204.

Aging impairs tendon stem/progenitor cell function and tendon homeostasis, however, effective treatments for aging-induced tendon diseases are lacking. Exosomes are naturally derived nanoparticles that contain bioactive molecules, and therefore, have attracted great interest in tissue engineering and regenerative medicine. In this study, it is shown that young exosomes secreted by stem cells from human exfoliated deciduous teeth (SHED-Exos) possess abundant anti-aging signals. These young bio-nanoparticles can alleviate the aging phenotypes of aged tendon stem/progenitor cells (AT-SCs) and maintain their tenogenic capacity. Mechanistically, SHED-Exos modulate histone methylation and inhibit nuclear factor-κB to reverse AT-SC aging. In a naturally aging mouse model, systemic administration of SHED-Exo bio-nanoparticles retards tendon degeneration. Interestingly, local delivery of SHED-Exos-loaded microspheres confers anti-aging phenotypes, including reduced senescent cells and decreased ectopic bone formation, thereby functionally and structurally rescuing endogenous tendon regeneration and repair capacity in aged rats. Overall, SHED-Exos, as natural bioactive nanoparticles, have promising translational and therapeutic potential for aging-related diseases.

Current strategies for engineering bladder tissues include a bladder biopsy for in vitro cell expansion for use in reconstructive procedures. However, this approach cannot be used in patients with bladder cancer who need a complete bladder replacement. Bone marrow mesenchymal stem cells (BMSC) might be an alternative cell source to better meet this need. We investigated the effects of soluble growth factors, bladder extracellular matrix (ECM), and 3D dynamic culture on cell proliferation and differentiation of human BMSC into smooth muscle cells (SMC). Myogenic growth factors (PDGF-BB and TGF-β1) alone, or combined either with bladder ECM or dynamic cultures, induced BMSC to express smooth muscle-specific genes and proteins. Either ECM or the dynamic culture alone promoted cell proliferation but did not induce myogenic differentiation of BMSC. A highly porous poly-l-lactic acid (PLLA) scaffold provided a 3D structure for maximizing the cell-matrix penetration, maintained myogenic differentiation of the induced BMSC, and promoted tissue remolding with rich capillary formation in vivo. Our results demonstrate that myogenic-differentiated BMSC seeded on a nano fibrous PLLA scaffold can be potentially used for cell-based tissue engineering for bladder cancer patients requiring cystoplasty.

Mesenchymal stromal cells (MSCs) are a promising therapy for preventing chronic Graft-Versus-Host Disease (cGVHD) due to their potent immunomodulatory properties. However, the safety concerns regarding the use of MSCs remain unsolved, and conflicting effects are observed due to the heterogeneity of MSCs. Recently, exosomes were shown to mediate the paracrine effects of MSCs, making it a potential candidate for cell-free therapies. The aim of this study is to investigate the efficacy and safety of MSCs-derived exosomes (MSCs-exo) in an established cGVHD mouse model.Bone marrow (BM)-derived MSCs were cultured, and the supernatants of these cultures were collected to prepare exosomes using ultracentrifugation. Exosomes from human dermal fibroblasts (Fib-exo) were used as a negative control. The cGVHD model was established, and tail vein injections of MSCs-exo or Fib-exo were administered once per week for 6 weeks. The symptoms and signs of cGVHD were monitored, and histopathological changes were detected by hematoxylin and eosin and Masson staining. The effects of MSCs-exo on Th17, Th1, and Treg were evaluated by flow cytometry, qPCR, and Luminex. In addition, human peripheral blood mononuclear cells (PBMCs) were stimulated and treated with MSCs-exo in vitro. IL-17-expressing Th17 and IL-10-expressing Treg were evaluated by flow cytometry, qPCR, and ELISA.We found that MSCs-exo effectively prolonged the survival of cGVHD mice and diminished the clinical and pathological scores of cGVHD. Fibrosis in the skin, lung, and liver was significantly ameliorated by MSCs-exo application. In MSCs-exo treated mice, activation of CD4+ T cells and their infiltration into the lung were reduced. Of note, MSCs-exo exhibited potent immunomodulatory effects via the inhibition of IL-17-expressing pathogenic T cells and induction of IL-10-expressing regulatory cells during cGVHD. The expressions of Th17 cell-relevant transcription factors and pro-inflammatory cytokines was markedly reduced after MSCs-exo treatment. In vitro, MSCs-exo blocked Th17 differentiation and improved the Treg phenotype in PBMCs obtained from healthy donors and patients with active cGVHD, further indicating the regulatory effect of MSCs-exo on GVHD effector T cells.Our data suggested that MSCs-exo could improve the survival and ameliorate the pathologic damage of cGVHD by suppressing Th17 cells and inducing Treg. This finding provides a novel alternative approach for the treatment of cGVHD.

Abstract Aflatoxin B1 (AFB1) is known as a mycotoxin that causes various health problems in animals, but the precise mechanism of AFB1 on mitochondrial functions and apoptosis in primary broiler hepatocytes (PBHs) is not clear. The objective of this study was to investigate the effects of AFB1 on the mitochondrial functions, reactive oxygen species (ROS) generation, apoptosis and nuclear factor erythroid 2‐like factor 2 (Nrf2)‐related signal pathway in PBHs. Here, the mitochondrial membrane potential (MMP), ROS generation, antioxidative genes and apoptosis in PBHs induced by AFB1 were investigated. The results showed that AFB1 evoked mitochondrial ROS generation, decreased MMP and induced apoptosis in PBHs. AFB1 increased the percentage of apoptotic cells, and expression of caspase‐9 and caspase‐3, upregulated messenger RNA (mRNA) expression of Nrf2 and downregulated mRNA expressions of NAD(P)H: quinine oxidoreductase 1, superoxide dismutase and Heme oxygenase 1 in PBHs. The expression of Bax was also observed in cytoplasm. These findings suggested AFB1 results in a significant impairment of mitochondrial functions, activates ROS generation, induces apoptosis, and is involved in Nrf2 signal pathway through mitochondria ROS‐dependent signal pathways in PBHs.

Background: Acute liver failure is an inflammation-mediated hepatocyte injury. Mesenchymal stem cell (MSC) transplantation is currently considered to be an effective treatment strategy for acute liver failure. Exosomes are an important paracrine factor that can be used as a direct therapeutic agent. However, the use of bone marrow mesenchymal stem cell-derived exosomes (BMSC-Exos) in the treatment of acute liver failure has not been reported. Purpose: Here, we established a model of hepatocyte injury and apoptosis induced by D-galactosamine and lipopolysaccharide (D-GalN/LPS) to study the protective effect of BMSC-Exos on hepatocyte apoptosis, and further explored its protective mechanism. Methods: BMSC-Exos was identified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blot. Laser confocal microscopy was used to observe the uptake of Dil-Exos by hepatocytes. D-GalN/LPS-induced primary hepatocytes were pretreated with BMSC-Exos in vitro, and then the cells were harvested. The apoptosis of hepatocytes was observed by TUNEL staining, flow cytometry and Western blot. Electron microscopy and mRFP-GFP-LC3 and Western blot was used to observe autophagy. Results: BMSC-Exos increased the expression of autophagy marker proteins LC3 and Beclin-1 and promoted the formation of autophagosomes. After BMSC-Exos treatment, the expression levels of the proapoptotic proteins Bax and cleaved caspase-3 were significantly decreased, while the expression level of the anti-apoptotic protein Bcl-2 was upregulated. However, when the autophagy inhibitor 3MA was present, the effect of BMSC-Exos on inhibiting apoptosis was significantly reversed. Conclusions: Our results showed for the first time that BMSC-Exos had the potential to reduce hepatocyte apoptosis after acute liver failure. In particular, we found that BMSC-Exos attenuated hepatocyte apoptosis by promoting autophagy. Keywords: bone marrow mesenchymal stem cells, exosomes, D-GalN/LPS, apoptosis, autophagy, acute liver failure

Titanium (Ti) and its alloys offer favorable biocompatibility, mechanical properties and corrosion resistance, which makes them an ideal material choice for dental implants. However, the long-term success of Ti-based dental implants may be challenged due to implant-related infections and inadequate osseointegration. With the development of nanotechnology, nanoscale modifications and the application of nanomaterials have become key areas of focus for research on dental implants. Surface modifications and the use of various coatings, as well as the development of the controlled release of antibiotics or proteins, have improved the osseointegration and soft-tissue integration of dental implants, as well as their antibacterial and immunomodulatory functions. This review introduces recent nano-engineering technologies and materials used in topographical modifications and surface coatings of Ti-based dental implants. These advances are discussed and detailed, including an evaluation of the evidence of their biocompatibility, toxicity, antimicrobial activities and in-vivo performances. The comparison between these attempts at nano-engineering reveals that there are still research gaps that must be addressed towards their clinical translation. For instance, customized three-dimensional printing technology and stimuli-responsive, multi-functional and time-programmable implant surfaces holds great promise to advance this field. Furthermore, long-term in vivo studies under physiological conditions are required to ensure the clinical application of nanomaterial-modified dental implants.

Post-extraction bleeding and alveolar bone resorption are the two frequently encountered complications after tooth extraction that result in poor healing and rehabilitation difficulties. The present study covalently bonded polyphosphate onto a collagen scaffold (P-CS) by crosslinking. The P-CS demonstrated improved hemostatic property in a healthy rat model and an anticoagulant-treated rat model. This improvement is attributed to the increase in hydrophilicity, increased thrombin generation, platelet activation and stimulation of the intrinsic coagulation pathway. In addition, the P-CS promoted the in-situ bone regeneration and alveolar ridge preservation in a rat alveolar bone defect model. The promotion is attributed to enhanced osteogenic differentiation of bone marrow stromal cells. Osteogenesis was improved by both polyphosphate and blood clots. Taken together, P-CS possesses favorable hemostasis and alveolar ridge preservation capability. It may be used as an effective treatment option for post-extraction bleeding and alveolar bone loss.Collagen scaffold is commonly used for the treatment of post-extraction bleeding and alveolar bone loss after tooth extraction. However, its application is hampered by insufficient hemostatic and osteoinductive property. Crosslinking polyphosphate with collagen produces a modified collagen scaffold that possesses improved hemostatic performance and augmented bone regeneration potential.

Glucose metabolism plays a pivotal role in many physiological and pathological conditions. To investigate the effect of hypoglycemia (obtained by glucose deprivation) on PC12 cell line, we analyzed the cell viability, mitochondrial function (assessed by MTT reduction, cellular ATP level, mitochondrial transmembrane potential), and the level of reactive oxygen species (ROS) after glucose deprivation (GD). Upon exposure to GD, ROS level increased and MTT reduction decreased immediately, intracellular ATP level increased in the first 3 hours, followed by progressive decrease till the end of GD treatment, and the mitochondrial transmembrane potential (deltapsi(m)) dropped after 6 hours. Both necrosis and apoptosis occurred apparently after 24 hours which was determined by nuclei staining with propidium iodide(PI) and Hoechst 33342. These data suggested that cytotoxicity of GD is mainly due to ROS accumulation and ATP depletion in PC12 cells.

We recently reported that TNFR-associated factor (TRAF)3, a ubiquitously expressed adaptor protein, promotes mature B cell apoptosis. However, the specific function of TRAF3 in T cells has remained unclear. In this article, we report the generation and characterization of T cell-specific TRAF3(-/-) mice, in which the traf3 gene was deleted from thymocytes and T cells. Ablation of TRAF3 in the T cell lineage did not affect CD4 or CD8 T cell populations in secondary lymphoid organs or the numbers or proportions of CD4(+),CD8(+) or double-positive or double-negative thymocytes, except that the T cell-specific TRAF3(-/-) mice had a 2-fold increase in FoxP3(+) T cells. In striking contrast to mice lacking TRAF3 in B cells, the T cell TRAF3-deficient mice exhibited defective IgG1 responses to a T-dependent Ag, as well as impaired T cell-mediated immunity to infection with Listeria monocytogenes. Surprisingly, we found that TRAF3 was recruited to the TCR/CD28 signaling complex upon costimulation and that TCR/CD28-mediated proximal and distal signaling events were compromised by TRAF3 deficiency. These findings provide insights into the roles played by TRAF3 in T cell activation and T cell-mediated immunity.

According to traditional Chinese medicine (TCM) theory, acupoints are specifically chosen sites of acupuncture manipulation, and also the basis for studying the mechanism of acupuncture. Stimulating different acupoints on the body surface could provide various therapeutic benefits. However, what is the acupoint? This question is not clear.We focuse on examining the function of acupoints from different perspectives, including the local and the systemic effects of stimulating acupoints. For example, acupoints may release certain substances or incur some changes, which could adjust the function of organs, maintain homeostasis. Furthermore, the therapeutic effects of verum acupoints versus sham acupoints were discussed. However, due to insufficience in evidence and in current methodologies, research into mechanisms of acupuncture is still incomplete.This review might explain, to some extent, what an acupoint is. Further research into the identity of acupoints is warranted, and multidisciplinary methods using novel technologies may yield significant advances over existing knowledge.

Background: DaxibotulinumtoxinA for Injection (DAXI) is a novel botulinum toxin type A formulation in clinical development. A phase 2 dose-ranging study identified an optimal dose and demonstrated efficacy with a median duration of 24 weeks. Methods: In two phase 3, multicenter, randomized, double-blind, placebo-controlled studies (SAKURA 1 and SAKURA 2), subjects with moderate or severe glabellar lines at maximum frown were assigned randomly to receive placebo or 40 U of DAXI. Glabellar lines were evaluated at least every 4 weeks for at least 24 weeks until severity returned to baseline (≤36 weeks). Results: Overall, 609 subjects were enrolled (DAXI, n = 405; placebo, n = 204). DAXI was significantly more effective than placebo in achieving the primary efficacy outcome (≥2-point improvement in glabellar line severity at maximum frown at week 4 according to both investigator and subject ratings): 73.6 percent versus 0.0 percent (SAKURA 1), and 74.0 percent versus 1.0 percent (SAKURA 2) (both p < 0.0001). Composite investigator and subject ratings of maximum frown after DAXI treatment showed that glabellar line severity of none or mild was maintained for a median of 24.0 weeks (SAKURA 1) and 23.9 weeks (SAKURA 2), and glabellar line severity did not return to baseline levels for a median of 27.7 and 26.0 weeks, respectively. DAXI was generally well tolerated, with the most common adverse events related to DAXI treatment being headache (SAKURA 1, 7.0 percent; SAKURA 2, 5.9 percent) and injection-site pain (5.0 percent and 2.4 percent, respectively). Conclusions: Results from both studies were highly consistent. DAXI may offer a prolonged duration of response (median, ≥24 weeks) and is generally well tolerated. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, I.

Mechanical force plays an important role in modulating stem cell fate and behaviours. However, how periodontal ligament stem cells (PDLSCs) perceive mechanical stimulus and transfer it into biological signals, and thereby promote alveolar bone remodelling, is unclear.An animal model of force-induced tooth movement and a compressive force in vitro was used. After force application, tooth movement distance, mesenchymal stem cell and osteoclast number, and proinflammatory cytokine expression were detected in periodontal tissues. Then, rat primary PDLSCs with or without force loading were isolated, and their stem cell characteristics including clonogenicity, proliferation, multipotent differentiation and immunoregulatory properties were evaluated. Under compressive force in vitro, the effects of the ERK signalling pathway on PDLSC characteristics were evaluated by Western blotting.Mechanical force in vivo induced PDLSC proliferation, which was accompanied with inflammatory cytokine accumulation, osteoclast differentiation and TRPV4 activation; the force-stimulated PDLSCs showed greater clonogenicity and proliferation, reduced differentiation ability, improved induction of macrophage migration, osteoclast differentiation and proinflammatory factor expression. The biological changes induced by mechanical force could be partially suppressed by TRPV4 inhibition. Mechanistically, force-induced activation of TRPV4 in PDLSCs regulated osteoclast differentiation by affecting the RANKL/OPG system via ERK signalling.Taken together, we show here that TRPV4 activation in PDLSCs under mechanical force contributes to changing their stem cell characteristics and modulates bone remodelling during tooth movement.

Extensive bone defect repair remains a clinical challenge, since ideal implantable scaffolds require the integration of excellent biocompatibility, sufficient mechanical strength and high biological activity to support bone regeneration. The inorganic nanomaterial-based therapy is of great significance due to their excellent mechanical properties, adjustable biological interface and diversified functions. Calcium–phosphorus compounds, silica and metal-based materials are the most common categories of inorganic nanomaterials for bone defect repairing. Nano hydroxyapatites, similar to natural bone apatite minerals in terms of physiochemical and biological activities, are the most widely studied in the field of biomineralization. Nano silica could realize the bone-like hierarchical structure through biosilica mineralization process, and biomimetic silicifications could stimulate osteoblast activity for bone formation and also inhibit osteoclast differentiation. Novel metallic nanomaterials, including Ti, Mg, Zn and alloys, possess remarkable strength and stress absorption capacity, which could overcome the drawbacks of low mechanical properties of polymer-based materials and the brittleness of bioceramics. Moreover, the biodegradability, antibacterial activity and stem cell inducibility of metal nanomaterials can promote bone regeneration. In this review, the advantages of the novel inorganic nanomaterial-based therapy are summarized, laying the foundation for the development of novel bone regeneration strategies in future.

Nano-engineering-based tissue regeneration and local therapeutic delivery strategies show significant potential to reduce the health and economic burden associated with craniofacial defects, including traumas and tumours. Critical to the success of such nano-engineered non-resorbable craniofacial implants include load-bearing functioning and survival in complex local trauma conditions. Further, race to invade between multiple cells and pathogens is an important criterion that dictates the fate of the implant. In this pioneering review, we compare the therapeutic efficacy of nano-engineered titanium-based craniofacial implants towards maximised local therapy addressing bone formation/resorption, soft-tissue integration, bacterial infection and cancers/tumours. We present the various strategies to engineer titanium-based craniofacial implants in the macro-, micro- and nano-scales, using topographical, chemical, electrochemical, biological and therapeutic modifications. A particular focus is electrochemically anodised titanium implants with controlled nanotopographies that enable tailored and enhanced bioactivity and local therapeutic release. Next, we review the clinical translation challenges associated with such implants. This review will inform the readers of the latest developments and challenges related to therapeutic nano-engineered craniofacial implants.

Abstract Osteoinductive synthetic biomaterials for replacing autografts can be developed by mimicking bone hierarchy and surface topography for host cell recruitment and differentiation. Until now, it has been challenging to reproduce a bone‐like staggered hierarchical structure since the energy change underlying synthetic pathways in vitro is essentially different from that of the natural process in vivo. Herein, a bone‐like hierarchically staggered architecture is reproduced under thermodynamic control involving two steps: fabrication of a high‐energy polyacrylic acid‐calcium intermediate and selective mineralization in collagenous gap regions driven by an energetically downhill process. The intermediate energy interval could easily be adjusted to determine different mineralization modes, with distinct morphologies and biofunctions. Similar to bone autografts, the staggered architecture offers a bone‐specific microenvironment for stem cell recruitment and multidifferentiation in vitro, and induces neo‐bone formation with bone marrow blood vessels by host stem cell homing in vivo. This work provides a novel perspective for an in vitro simulating biological mineralization process and proof of concept for the clinical application of smart biomaterials.

Abstract Mineral granules in the mitochondria of bone‐forming cells are thought to be the origin of biomineral precursors, which are transported to extracellular matrices to initiate cell‐mediated biomineralization. However, no evidence has revealed how mitochondrial granules form. This study indicates that mitochondrial granules are initiated by transporting calcium and phosphorus clusters from the endoplasmic reticulum (ER) to mitochondria based on detailed observations of the continuous process of mouse parietal bone development as well as in vitro biomineralization in bone‐forming cells. Nanosized biomineral precursors (≈30 nm in diameter), which originate from mitochondrial granules, initiate intrafibrillar mineralization of collagen as early as embryonic day 14.5. Both in vivo and in vitro studies further reveal that formation of mitochondrial granules is induced by the ER. Elevated levels of intracellular calcium or phosphate ions, which can be induced by treatment with ionomycin and black phosphorus, respectively, accelerate formation of the calcium and phosphorus clusters on ER membranes and ultimately promote biomineralization. These findings provide a novel insight into biomineralization and bone formation.

Autophagy, a lysosomal degradation pathway, serves as a protective cellular mechanism in maintaining cell and tissue homeostasis under mechanical stimulation. As the mechanosensitive cells, periodontal ligament stem cells (PDLSCs) play an important role in the force-induced inflammatory bone remodeling and tooth movement process. However, whether and how autophagy in PDLSCs influences the inflammatory bone remodeling process under mechanical force stimuli is still unknown. In this study, we found that mechanical force stimuli increased the expression of the autophagy protein LC3, the number of M1 macrophages and osteoclasts, as well as the ratio of M1/M2 macrophages in the compression side of the periodontal ligament in vivo . These biological changes induced by mechanical force were repressed by the application of an autophagy inhibitor 3-methyladenine. Moreover, autophagy was activated in the force-loaded PDLSCs, and force-stimulated PDLSC autophagy further induced M1 macrophage polarization in vitro. The macrophage polarization could be partially blocked by the administration of autophagy inhibitor 3-methyladenine or enhanced by the administration of autophagy activator rapamycin in PDLSCs. Mechanistically, force-induced PDLSC autophagy promoted M1 macrophage polarization via the inhibition of the AKT signaling pathway. These data suggest a novel mechanism that force-stimulated PDLSC autophagy steers macrophages into the M1 phenotype via the AKT signaling pathway, which contributes to the inflammatory bone remodeling and tooth movement process.

Macrophages are involved mainly in the balance between inflammation and tenogenesis during the healing process of tendinopathy. However, etiological therapeutic strategies to efficiently treat tendinopathy by modulating macrophage state are still lacking. In this study, we find that a small molecule compound Parishin-A (PA) isolated from Gastrodia elata could promote anti-inflammatory M2 macrophage polarization by inhibiting gene transcription and protein phosphorylation of signal transducers and activators of transcription 1. Local injection or sustained delivery of PA by mesoporous silica nanoparticles (MSNs) could almost recover the native tendon's dense parallel-aligned collagen matrix in collagenase-induced tendinopathy by modulating macrophage-mediated immune microenvironment and preventing heterotopic ossification. Especially, MSNs decrease doses of PA, frequency of injection and yield preferable therapeutic effects. Mechanistically, intervention with PA could indirectly inhibit activation of mammalian target of rapamycin to repress chondrogenic and osteogenic differentiation of tendon stem/progenitor cells by influencing macrophage inflammatory cytokine secretion. Together, pharmacological intervention with a natural small-molecule compound to modulate macrophage status appears to be a promising strategy for tendinopathy treatment.

Infected bone defects are a major challenge in orthopedic treatment. Native bone tissue possesses an endogenous electroactive interface that induces stem cell differentiation and inhibits bacterial adhesion and activity. However, traditional bone substitutes have difficulty in reconstructing the electrical environment of bone. In this study, we develop a self-promoted electroactive mineralized scaffold (sp-EMS) that generates weak currents via spontaneous electrochemical reactions to activate voltage-gated Ca2+ channels, enhance adenosine triphosphate-induced actin remodeling, and ultimately achieve osteogenic differentiation of mesenchymal stem cells by activating the BMP2/Smad5 pathway. Furthermore, we show that the electroactive interface provided by the sp-EMS inhibits bacterial adhesion and activity via electrochemical products and concomitantly generated reactive oxygen species. We find that the osteogenic and antibacterial dual functions of the sp-EMS depend on its self-promoting electrical stimulation. We demonstrate that in vivo, the sp-EMS achieves complete or nearly complete in situ infected bone healing, from a rat calvarial defect model with single bacterial infection, to a rabbit open alveolar bone defect model and a beagle dog vertical bone defect model with the complex oral bacterial microenvironment. This translational study demonstrates that the electroactive bone graft presents a promising therapeutic platform for complex defect repair.

"Lisfranc joint injury" is comprised of a tarsometatarsal joint-complex injury. The Lisfranc complex injury is always a challenge for orthopedists, and the optimum treatment is still up for debate. Anatomic reduction and stable internal fixation prove to have no satisfactory outcomes. This research aims to compare the clinical curative effects, complications and radiographic features of arthrodesis and non-fusion of the Lisfranc joint in the follow-up of the patients who suffered Lisfranc injuries.A comparative retrospective study of 25 patients with acute or subacute Lisfranc complex injuries was conducted between September 2013 and March 2015 in the First Affiliated Hospital of Soochow University. All patients were classified by Myerson classification. Eight patients were treated with arthrodesis, while 17 patients received non-fusion operations. The clinical curative effects, complications and image differences were compared between the two groups. American Orthopaedic Foot and Ankle Society (AOFAS) hindfoot score, Short Form-36 (SF-36) and Visual Analogue Scale (VAS) score were evaluated for each patient during the follow-up. All statistics were analyzed using the SPSS software system.All fractures healed for both the arthrodesis group and the non-fusion group. Patients in the arthrodesis group had a higher AOFAS score compared with patients in the non-fusion group (94.00 vs. 88.58, P = 0.034). Complications occurred in eight patients (8/17, 47%) in the non-arthrodesis group, including the second and third phalanx abduction (1), talipes cavus (2), eversion deformity of front foot (3), eversion deformity of calcaneus (1), as well as postoperative infection (1). Only two patients (2/8, 25%) in the arthrodesis group suffered complications. One was a limitation of motion of the front foot and pain during walking; the other was an eversion deformity of front foot.Primary arthrodesis has advantages compared to primary open reduction and internal fixation (ORIF): reduced foot deformity rates, sustained biomechanical morphology of the feet, reduced complications, higher level of function recovery, shorter time of surgical procedures, fewer complications, higher AOFAS score and fewer frequency of complications. According to our research, primary arthrodesis may be a better choice for treating Lisfranc injury.

Abstract Biomimetic intrafibrillarly-mineralized collagen (IMC) is a promising scaffold for bone regeneration because of its structural and functional similarity to natural bone. The objective of this study was to evaluate the bone regeneration potential of IMC loaded with autologous periodontal ligament stem cells (PDLSCs) in large bone defects in minipigs. A macroporous IMC with a bone-like subfibrillar nanostructure was fabricated using a biomimetic bottom-up approach. Non-healing full thickness defects were established on the cranial bone in minipigs, and IMC and hydroxyapatite (HA) scaffolds seeded with autologous PDLSCs were implanted into these defects. Computed tomographic imaging, histology staining, and atomic force microscopy were applied to evaluate to the quantity, micro/nano structures, and mechanical performance of the neo-bone after 12 weeks of implantation. Compared with HA, IMC showed superior regeneration properties characterized by the profuse deposition of new bony structures with a normal architecture and vascularization. Immunohistochemistry showed that the runt-related transcription factor 2 and transcription factor Osterix were highly expressed in the neo-bone formed by IMC. Furthermore, the nanostructure and nanomechanics of the neo-bone formed by IMC were similar to that of natural bone. This study provides strong evidence for the future clinical applications of the IMC-based bone grafts.

Reactive oxygen species (ROS) contribute to cellular redox environment and serve as signaling molecules. Excessive ROS can lead to oxidative stress that are involved in a broad spectrum of physiological and pathological conditions. Stem cells have unique ROS regulation while cancer cells frequently show a constitutive oxidative stress that is associated with the invasive phenotype. Antioxidants have been proposed to forestall tumor progression while targeted oxidants have been used to destroy tumor cells. However, the delicate beneficial range of ROS levels for stem cells and tumor cells under distinct contexts remains elusive. Here, we used Drosophila midgut intestinal stem cell (ISCs) as an in vivo model system to tackle this question. The ROS levels of ISCs remained low in comparison to that of differentiated cells and increased with ageing, which was accompanied by elevated proliferation of ISCs in aged Drosophila. Neither upregulation nor downregulation of ROS levels significantly affected ISCs, implicating an intrinsic homeostatic range of ROS in ISCs. Interestingly, we observed similar moderately elevated ROS levels in both tumor-like ISCs induced by Notch (N) depletion and extracellular matrix (ECM)-deprived ISCs induced by β-integrin (mys) depletion. Elevated ROS levels further promoted the proliferation of tumor-like ISCs while reduced ROS levels suppressed the hyperproliferation phenotype; on the other hand, further increased ROS facilitated the survival of ECM-deprived ISCs while reduced ROS exacerbated the loss of ECM-deprived ISCs. However, N- and mys-depleted ISCs, which resembled metastatic tumor cells, harbored even higher ROS levels and were subjected to more severe cell loss, which could be partially prevented by ectopic supply of antioxidant enzymes, implicating a delicate pro-surviving and proliferating range of ROS levels for ISCs. Taken together, our results revealed stem cells can differentially respond to distinct ROS levels under various conditions and suggested that the antioxidant-based intervention of stem cells and tumors should be formulated with caution according to the specific situations.

Titania nanotubes (TNTs) fabricated on titanium orthopedic and dental implants have shown significant potential in "proof of concept" in vitro, ex vivo, and short-term in vivo studies. However, most studies do not focus on a clear direction for future research towards clinical translation, and there exists a knowledge gap in identifying key research challenges that must be addressed to progress to the clinical setting. This review focuses on such challenges with respect to anodized titanium implants modified with TNTs, including optimized fabrication on clinically utilized microrough surfaces, clinically relevant bioactivity assessments, and controlled/tailored local release of therapeutics. Further, long-term in vivo investigations in compromised animal models under loading conditions are needed. We also discuss and detail challenges and progress related to the mechanical stability of TNT-based implants, corrosion resistance/electrochemical stability, optimized cleaning/sterilization, packaging/aging, and nanotoxicity concerns. This extensive, clinical translation focused review of TNTs modified Ti implants aims to foster improved understanding of key research gaps and advances, informing future research in this domain.

Hematopoietic stem cells (HSCs) can remain quiescent or they can enter the cell cycle, and either self-renew or differentiate. Although cyclin C and cyclin dependent kinase (cdk3) are essential for the transition from the G(0) to the G(1) phase of the cell cycle in human fibroblasts, the role of cyclin C in hematopoietic stem/progenitor cells (HSPCs) is not clear. We have identified an important role of cyclin C (CCNC) in regulating human HSPC quiescence, as knocking down CCNC expression in human cord blood CD34(+) cells resulted in a significant increase in quiescent cells that maintain CD34 expression. CCNC knockdown also promotes in vitro HSPC expansion and enhances their engraftment potential in sublethally irradiated immunodeficient mice. Our studies establish cyclin C as a critical regulator of the G(0)/G(1) transition of human HSPCs and suggest that modulating cyclin C levels may be useful for HSC expansion and more efficient engraftment.

ZG16p is a soluble mammalian lectin, the first to be described with a Jacalin-related β-prism-fold. ZG16p has been reported to bind both to glycosaminoglycans and mannose. To determine the structural basis of the multiple sugar-binding properties, we conducted glycan microarray analyses of human ZG16p. We observed that ZG16p preferentially binds to α-mannose-terminating short glycans such as Ser/Thr-linked O-mannose, but not to high mannose-type N-glycans. Among sulfated glycosaminoglycan oligomers examined, chondroitin sulfate B and heparin oligosaccharides showed significant binding. Crystallographic studies of human ZG16p lectin in the presence of selected ligands revealed the mechanism of multiple sugar recognition. Manα1–3Man and Glcβ1–3Glc bound in different orientations: the nonreducing end of the former and the reducing end of the latter fitted in the canonical shallow mannose binding pocket. Solution NMR analysis using 15N-labeled ZG16p defined the heparin-binding region, which is on an adjacent flat surface of the protein. On-array competitive binding assays suggest that it is possible for ZG16p to bind simultaneously to both types of ligands. Recognition of a broad spectrum of ligands by ZG16p may account for the multiple functions of this lectin in the formation of zymogen granules via glycosaminoglycan binding, and in the recognition of pathogens in the digestive system through α-mannose-related recognition. ZG16p is a soluble mammalian lectin, the first to be described with a Jacalin-related β-prism-fold. ZG16p has been reported to bind both to glycosaminoglycans and mannose. To determine the structural basis of the multiple sugar-binding properties, we conducted glycan microarray analyses of human ZG16p. We observed that ZG16p preferentially binds to α-mannose-terminating short glycans such as Ser/Thr-linked O-mannose, but not to high mannose-type N-glycans. Among sulfated glycosaminoglycan oligomers examined, chondroitin sulfate B and heparin oligosaccharides showed significant binding. Crystallographic studies of human ZG16p lectin in the presence of selected ligands revealed the mechanism of multiple sugar recognition. Manα1–3Man and Glcβ1–3Glc bound in different orientations: the nonreducing end of the former and the reducing end of the latter fitted in the canonical shallow mannose binding pocket. Solution NMR analysis using 15N-labeled ZG16p defined the heparin-binding region, which is on an adjacent flat surface of the protein. On-array competitive binding assays suggest that it is possible for ZG16p to bind simultaneously to both types of ligands. Recognition of a broad spectrum of ligands by ZG16p may account for the multiple functions of this lectin in the formation of zymogen granules via glycosaminoglycan binding, and in the recognition of pathogens in the digestive system through α-mannose-related recognition.

Abstract While Polycomb group protein Bmi1 is important for stem cell maintenance, its role in lineage commitment is largely unknown. We have identified Bmi1 as a novel regulator of erythroid development. Bmi1 is highly expressed in mouse erythroid progenitor cells and its deficiency impairs erythroid differentiation. BMI1 is also important for human erythroid development. Furthermore, we discovered that loss of Bmi1 in erythroid progenitor cells results in decreased transcription of multiple ribosomal protein genes and impaired ribosome biogenesis. Bmi1 deficiency stabilizes p53 protein, leading to upregulation of p21 expression and subsequent G0/G1 cell cycle arrest. Genetic inhibition of p53 activity rescues the erythroid defects seen in the Bmi1 null mice, demonstrating that a p53-dependent mechanism underlies the pathophysiology of the anemia. Mechanistically, Bmi1 is associated with multiple ribosomal protein genes and may positively regulate their expression in erythroid progenitor cells. Thus, Bmi1 promotes erythroid development, at least in part through regulating ribosome biogenesis. Ribosomopathies are human disorders of ribosome dysfunction, including Diamond-Blackfan anemia (DBA) and 5q− syndrome, in which genetic abnormalities cause impaired ribosome biogenesis, resulting in specific clinical phenotypes. We observed that BMI1 expression in human hematopoietic stem and progenitor cells from patients with DBA is correlated with the expression of some ribosomal protein genes, suggesting that BMI1 deficiency may play a pathological role in DBA and other ribosomopathies. Stem Cells 2015;33:925–938

Mesenchymal stem cells (MSCs) are a reliable source for cell-based regenerative medicine owing to their multipotency and biological functions. However, aging-induced systemic homeostasis disorders in vivo and cell culture passaging in vitro induce a functional decline of MSCs, switching MSCs to a senescent status with impaired self-renewal capacity and biased differentiation tendency. MSC functional decline accounts for the pathogenesis of many diseases and, more importantly, limits the large-scale applications of MSCs in regenerative medicine. Growing evidence implies that epigenetic mechanisms are a critical regulator of the differentiation programs for cell fate and are subject to changes during aging. Thus, we here review epigenetic dysregulations that contribute to MSC aging and osteoporosis. Comprehending detailed epigenetic mechanisms could provide us with a novel horizon for dissecting MSC-related pathogenesis and further optimizing MSC-mediated regenerative therapies.

LAB (linker for activation of B cells), also known as NTAL (non-T cell activation linker), is a LAT (linker for activation of T cells)-like adaptor protein that is expressed in B, NK, and mast cells. Its role in lymphocytes has not been clearly demonstrated. Here, we showed that aged LAB-deficient (Lat2(-/-)) mice developed an autoimmune syndrome. Lat2(-/-) T cells were hyperactivated and produced more cytokines than Lat2(+/+) T cells. Even though LAB was absent in naive T cells, LAB could be detected in activated Lat2(+/+) T cells. LAT-mediated signaling events were enhanced in Lat2(-/-) T cells; however, they were suppressed in T cells that overexpressed LAB. Mice with the Lat2 gene conditionally deleted from T cells also developed the autoimmune syndrome like Lat2(-/-) mice. Together, these data demonstrated an important role of LAB in limiting autoimmune response and exposed a mechanism regulating T cell activation.

Abstract The isonymy structure of 1.28 billion people registered in China's National Citizen Identity Information System was studied at the provincial, prefectural, and county administrative division levels. The isonymy was 0.026 for China as a whole. The average value of isonymy was 0.033 for the 30 provinces, 0.035 for the 334 prefectures, and 0.040 for the 2811 counties. The isonymy in China was much higher than in other countries. This finding may be partly explained by the low number of surnames in the Chinese language. Two regional features can be identified from the geographic distributions of isonymy. One feature is that the middle and lower reaches of the Yangtze River had the lowest values of isonymy at both the provincial and county levels. The second feature is that most counties with the highest values of isonymy were distributed in the provinces with high proportions of ethnic minorities. According to the dendrogram of surname distances, several clusters could be identified. Most provinces in a cluster were conterminous with one another. The one exception could be explained by demic migration called “braving the journey to the northeast of China.” Isolation by distance could be detected because the correlation coefficients between Nei's distance and the geographic distances at the provincial, prefectural, and county levels were 0.64, 0.43, and 0.37, respectively. Human behaviors in Chinese history that may have caused these results have been discussed, including cultural origin, migration, residential patterns, and ethnic distribution. Am J Phys Anthropol 148:341–350, 2012. © 2012 Wiley Periodicals, Inc.

This study was designed to assess the use of a 3D printing technique in total hip arthroplasty (THA) for severe hip deformities, where new and improved approaches are needed. THAs were performed from January 2015 to December 2016. Bioprosthesis artificial hip joints were used in both conventional and 3D printing hip arthroplasties. A total of 74 patients (57 cases undergoing conventional hip replacements and 17 undergoing 3D printing hip replacements) were followed‑up for an average of 24 months. The average age of the patients was 62.7 years. Clinical data between the patients treated with different approaches were compared. Results showed that the time to postoperative weight bearing and the Harris scores of the patients in the 3D printing group were better than those for patients in the conventional hip replacement group. Unfortunately, the postoperative infection and loosening rates were higher in the 3D printing group. However, there were no significant differences in femoral neck anteversion, neck shaft, acetabular or sharp angles between ipsilateral and contralateral sides in the 3D printing group (P>0.05). The femoral neck anteversion angle was significantly different between the two sides in the conventional hip replacement group (P<0.05). Based on these results, we suggest that the 3D printing approach provides a better short‑term curative effect that is more consistent with the physiological structure and anatomical characteristics of the patient, and we anticipate that its use will help improve the lives of many patients.

Objective To evaluate the accuracy, reliability, and efficiency of voxel- and surface-based registrations for cone-beam computed tomography (CBCT) mandibular superimposition in adult orthodontic patients. Methods Pre- and post-orthodontic treatment CBCT scans of 27 adult patients were obtained. Voxel- and surface-based CBCT mandibular superimpositions were performed using the mandibular basal bone as a reference. The accuracy of the two methods was evaluated using the absolute mean distance measured. The time that was required to perform the measurements using these methods was also compared. Statistical differences were determined using paired t-tests, and inter-observer reliability was assessed by intraclass correlation coefficients (ICCs). Results The absolute mean distance on seven mandible surface areas between voxel- and surface-based registrations was similar but not significantly different. ICC values of the surface-based registration were 0.918 to 0.990, which were slightly lower than those of voxel-based registration that ranged from 0.984 to 0.996. The time required for voxel-based registration and surface-based registration was 44.6 ± 2.5 s and 252.3 ± 7.1 s, respectively. Conclusions Both methods are accurate and reliable and not significantly different from each other. However, voxel-based registration is more efficient than surface-based registration for CBCT mandibular superimposition.

ZG16p is a soluble mammalian lectin that interacts with mannose and heparan sulfate. Here we describe detailed analysis of the interaction of human ZG16p with mycobacterial phosphatidylinositol mannosides (PIMs) by glycan microarray and NMR. Pathogen-related glycan microarray analysis identified phosphatidylinositol mono- and di-mannosides (PIM1 and PIM2) as novel ligand candidates of ZG16p. Saturation transfer difference (STD) NMR and transferred NOE experiments with chemically synthesized PIM glycans indicate that PIMs preferentially interact with ZG16p by using the mannose residues. The binding site of PIM was identified by chemical-shift perturbation experiments with uniformly (15)N-labeled ZG16p. NMR results with docking simulations suggest a binding mode of ZG16p and PIM glycan; this will help to elucidate the physiological role of ZG16p.

In-stent restenosis (ISR) is a complex disease that occurs after coronary stenting procedures. The development of quality materials and improvement of our understanding on significant factors regulating ISR are essential for enhancing prognosis. Vascular smooth muscle cells (VSMCs) are the main constituent cells of blood vessel walls, and dysfunction of VMSCs can exacerbate ISR. Accordingly, in this study, we explored the influence of wrinkled material topography on the biological functions of VSMCs.Polydimethylsiloxane with a wrinkled topography was synthesized using elastomer base and crosslinking and observed by atomic force microscopy. VSMC proliferation, apoptosis, and morphology were determined by Cell Counting Kit-8 assays, fluorescence-assisted cell sorting, and phalloidin staining. α-Smooth muscle actin (α-SMA), major histocompatibility complex (MHC), and calponin 1 (CNN-1) expression levels were measured by quantitative real-time polymerase chain reaction and western blotting. Moreover, p53 and cleaved caspase-3 expression levels were evaluated by western blotting in VSMCs to assess apoptotic induction.Surface topographies were not associated with a clear orientation or elongation of VSMCs. The number of cells was increased on wrinkled surfaces (0.7 μm in amplitude, and 3 μm in wavelength [W3]) compared with that on other surfaces, contributing to continuously increased cell proliferation. Moreover, interactions of VSMCs with the W3 surface suppressed phenotypic switching, resulting in ISR via regulation of α-SMA, calponin-1, and SM-MHC expression. The surface with an amplitude of 0.05 μm and a wavelength of 0.5 μm (W0.5) promoted apoptosis by inducing caspase 3 and p53 activities.Introduction of aligned topographies on biomaterial scaffolds could provide physical cues to modulate VSMC responses for engineering vascular constructs. Materials with wrinkled topographies could have applications in the development of stents to reduce ISR.

Vascular calcification (VC) has emerged as a key predictor of cardiovascular events in patients with chronic kidney disease (CKD). In recent years, an expanding body of research has put forth the concept of accelerated vascular aging among CKD patients, highlighting the significance of vascular cells senescence in the process of VC. Within the milieu of uremia, senescent vascular endothelial cells (VECs) release extracellular microvesicles (MV) that promote vascular smooth muscle cells (VSMCs) senescence, thereby triggering the subsequent osteogenic phenotypic switch and ultimately contributing to the VC process. In addition, senescent vascular progenitor or stem cells with diminished ability to differentiate into VECs and VSMCS, compromise the repair of vascular integrity, on the other hand, release a cascade of molecules associated with senescence, collectively known as the senescence-associated secretory phenotype (SASP), perpetuating the senescence phenomenon. Furthermore, SASP triggers the recruitment of monocytes and macrophages, as well as adjacent VECs and VSMCs into a pro-adhesive and pro-inflammatory senescent state. This pro-inflammatory microenvironment niche not only impacts the functionality of immune cells but also influences the differentiation of myeloid immune cells, thereby amplifying the reduced ability to effectively clear senescent cells of senescent macrophages, promoted calcification of VSMCs. The objective of this paper is to provide a comprehensive review of the contribution of vascular cell senescence to the emergence and advancement of VC. Gaining a comprehensive understanding of the involvement of cellular senescence within the vessel wall is pivotal, especially when it comes to its intersection with VC. This knowledge is essential for advancing groundbreaking anti-aging therapies, aiming to effectively mitigate cardiovascular diseases.

Background. Triptolide (TPT) is a component of the Chinese herb Triptergium wilfordii and has potent immunosuppressive and anti-inflammatory effects. Since dendritic cells (DCs) play a critical role in the initiation of alloimmune responses to foreign grafts, the aim of the present study was to evaluate the effects of TPT in the functional phenotype of bone marrow (BM)-derived DCs. Methods. BM-derived DCs were cultured with or without TPT from days 2 to 7 and then stimulated with lipopolysaccharide (LPS) for a further 2 days. In some experiments, TPT was added to the cultures from day 7 to day 9. Heterotopic cardiac transplantation was performed and animals received either no treatment or were injected systemically with different doses of TPT posttransplantation. Results. TPT treatment inhibited LPS-induced DC maturation. The ability of DCs to stimulate allogeneic T-cell responses was also impaired by TPT treatment and accompanied by upregulated synthesis of interleukin-10 facilitating the expansion of regulatory T cells. Furthermore, we observed that TPT treatment led to increased cell surface expression of DC-SIGN and reduced expression of TLR4 on DCs. Conclusions. These findings demonstrate that TPT can inhibit the maturation and allogenicity of DCs and promotes the expansion of regulatory T cells. These effects were confirmed by in vivo experiments though treatment of TPT can prolong mice heart allograft survival. Furthermore, modulation of the DC surface phenotype towards that of a tolerogenic phenotype could contribute to the ability of TPT to suppress productive immunity and maintain homeostasis.

TIGIT expression on natural killer (NK) cells is associated with dysfunction during chronic HIV infection, but the phenotype and biological functions of these cells in the context of acute HIV-1 infection remain poorly understood. Here, 19 acutely infected HIV-1 patients traced at first, third and twelfth month, and age-matched patients with chronic HIV-1 infection were enrolled to investigate the phenotype and functions of TIGIT expression on NK cells. We found that TIGIT-expressing NK cells did not increase in frequency in the first, third and twelfth month of infection until chronic HIV-1 infection lasted over 2 years. The number of TIGIT + NK cells in acute infection was positively associated with HIV-1 viral load ( r = 0.53, P = 0.0009). CD96 was significantly upregulated on NK cells after acute infection for 1 month and in chronic infection over 2 years, while CD226 was downregulated in chronic infection over 2 years. Further, at different stages of infection, CD96 − CD226 + cells diminished among total NK cells, TIGIT + NK and TIGIT − NK cells, while CD96 + CD226 − cells expanded. Reduced CD96 − CD226 + cells and elevated CD96 + CD226 − cells among NK cells especially TIGIT − NK cells, had opposite associations with viral load in the first month of infection, as well as CD4 T-cell counts in including the twelfth month and more than 2 years of chronic infection. In both HIV-1-infected individuals and healthy donors, TIGIT was predominantly expressed in NKG2A − NKG2C + NK cells, with a significantly higher proportion than in NKG2A + NKG2C − NK cells. Moreover, the frequencies of TIGIT + NK cells were positively associated with the frequencies of NKG2A − NKG2C + NK cells in acute infection ( r = 0.62, P &amp;lt; 0.0001), chronic infection ( r = 0.37, P = 0.023) and healthy donors ( r = 0.36, P = 0.020). Enhanced early activation and coexpression of CD38 and HLA-DR in TIGIT + NK cells were detected compared to TIGIT − NK cells, both of which were inversely associated with the decrease in CD4 T-cell counts in both acute and chronic HIV-1 infection. The ability of TIGIT + NK cells to produce TNF-α, IFN-γ and CD107a degranulation substance were consistently weaker than that of TIGIT − NK cells in both acute and chronic infection. Moreover, the functionalities of TIGIT + NK cells were lower than those of TIGIT − NK cells, except for TNF-α − CD107a + IFN-γ − NK cells. These findings highlight the phenotype and functional characteristics of TIGIT-expressing NK cells which have poor capabilities in inhibiting HIV-1 replication and maintaining CD4 T-cell counts.

Maintaining the stemness of bone marrow mesenchymal stem cells (BMMSCs) is crucial for bone homeostasis and regeneration. However, in vitro expansion and bone diseases impair BMMSC stemness, limiting its functionality in bone tissue engineering. Using a deep learning-based efficacy prediction system and bone tissue sequencing, we identify a natural small-molecule compound, dihydroartemisinin (DHA), that maintains BMMSC stemness and enhances bone regeneration. During long-term in vitro expansion, DHA preserves BMMSC stemness characteristics, including its self-renewal ability and unbiased differentiation. In an osteoporosis mouse model, oral administration of DHA restores the femur trabecular structure, bone density, and BMMSC stemness in situ. Mechanistically, DHA maintains BMMSC stemness by promoting histone 3 lysine 9 acetylation via GCN5 activation both in vivo and in vitro. Furthermore, the bone-targeted delivery of DHA by mesoporous silica nanoparticles improves its therapeutic efficacy in osteoporosis. Collectively, DHA could be a promising therapeutic agent for treating osteoporosis by maintaining BMMSC stemness.

Bone infection poses a major clinical challenge that can hinder patient recovery and exacerbate postoperative complications. This study has developed a bioactive composite scaffold through the co-assembly and intrafibrillar mineralization of collagen fibrils and zinc oxide (ZnO) nanowires (IMC/ZnO). The IMC/ZnO exhibits bone-like hierarchical structures and enhances capabilities for osteogenesis, antibacterial activity, and bacteria-infected bone healing. During co-cultivation with human bone marrow mesenchymal stem cells (BMMSCs), the IMC/ZnO improves BMMSC adhesion, proliferation, and osteogenic differentiation even under inflammatory conditions. Moreover, it suppresses the activity of Gram-negative Porphyromonas gingivalis and Gram-positive Streptococcus mutans by releasing zinc ions within the acidic infectious microenvironment. In vivo, the IMC/ZnO enables near-complete healing of infected bone defects within the intricate oral bacterial milieu, which is attributed to IMC/ZnO orchestrating M2 macrophage polarization, and fostering an osteogenic and anti-inflammatory microenvironment. Overall, these findings demonstrate the promise of the bioactive scaffold IMC/ZnO for treating bacteria-infected bone defects.

Abstract Pyroptosis, an inflammatory caspase-dependent programmed cell death, plays a vital role in maintaining tissue homeostasis and activating inflammatory responses. Orthodontic tooth movement (OTM) is an aseptic force-induced inflammatory bone remodeling process mediated by the activation of periodontal ligament (PDL) progenitor cells. However, whether and how force induces PDL progenitor cell pyroptosis, thereby influencing OTM and alveolar bone remodeling remains unknown. In this study, we found that mechanical force induced the expression of pyroptosis-related markers in rat OTM and alveolar bone remodeling process. Blocking or enhancing pyroptosis level could suppress or promote OTM and alveolar bone remodeling respectively. Using Caspase-1 −/− mice, we further demonstrated that the functional role of the force-induced pyroptosis in PDL progenitor cells depended on Caspase-1. Moreover, mechanical force could also induce pyroptosis in human ex-vivo force-treated PDL progenitor cells and in compressive force-loaded PDL progenitor cells in vitro, which influenced osteoclastogenesis. Mechanistically, transient receptor potential subfamily V member 4 signaling was involved in force-induced Caspase-1-dependent pyroptosis in PDL progenitor cells. Overall, this study suggested a novel mechanism contributing to the modulation of osteoclastogenesis and alveolar bone remodeling under mechanical stimuli, indicating a promising approach to accelerate OTM by targeting Caspase-1.

The spatial distribution of Chinese surnames is diverse and provides rich information about the evolution of human society. This study aims to propose several indices to quantify the spatial distribution characteristics of Chinese common surnames and to explore how these distributions are related to historical evolution.This study uses data from China's ID information system covering 1.28 billion people across 362 cities. Based on the location quotient, several new concepts, such as "moderately concentrated cities" and "highly concentrated cities," are defined. Then indices such as range, ununiformity and spatial autocorrelation are proposed and calculated to analyze the spatial characteristics of Chinese common surnames.A significant correlation is observed between the commonness of a surname and its spatial characteristics: the more common the surname, the wider its spatial range, the lower the ununiformity, and the higher the autocorrelation coefficient. These patterns reflect the complex interplay of historical, geographical, and cultural factors influencing surname spatial distribution.The spatial distribution of Chinese surnames is intricately linked to their historical evolution. Most common surnames, often with deeper historical roots, exhibit wider distributions and lower ununiformity, whereas less common surnames show higher concentrations in specific areas. These quantitative results provide a comprehensive understanding of the evolutionary characteristics of Chinese surnames.

Abstract Malocclusion, identified by the World Health Organization (WHO) as one of three major oral diseases, profoundly impacts the dental-maxillofacial functions, facial esthetics, and long-term development of ~260 million children in China. Beyond its physical manifestations, malocclusion also significantly influences the psycho-social well-being of these children. Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition, by mitigating the negative impact of abnormal environmental influences on the growth. Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development, ranging from fetal stages to the early permanent dentition phase. From an economic and societal standpoint, the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated, underlining its profound practical and social importance. This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children, emphasizing critical need for early treatment. It elaborates on corresponding core principles and fundamental approaches in early orthodontics, proposing comprehensive guidance for preventive and interceptive orthodontic treatment, serving as a reference for clinicians engaged in early orthodontic treatment.

Abstract The regenerative treatment of infectious vertical bone defects remains difficult and challenging today. Current clinical treatments are limited in their ability to control bacteria and infection, which is unfavorable for new bone formation and calls for a new type of material with excellent osteogenic and antibacterial properties. Here a multifunctional scaffold is synthesized that mimics natural bone nanostructures by incorporating silver nanowires into a hierarchical, intrafibrillar mineralized collagen matrix (IMC/AgNWs), to achieve the therapeutic goals of inhibiting bacterial activity and promoting infectious alveolar bone augmentation in rats and beagle dogs. An appropriate concentration of 0.5 mg mL −1 AgNWs is selected to balance biocompatibility and antibacterial properties. The achieved IMC/AgNWs exhibit a broad spectrum of antimicrobial properties against Gram‐negative Porphyromonas gingivalis and Gram‐positive Streptococcus mutans . When the IMC/AgNWs are cocultured with periodontal ligament stem cells, it possesses excellent osteoinductive activities under both non‐inflammatory and inflammatory conditions. By constructing a rat mandibular infected periodontal defect model, the IMC/AgNWs achieve a near‐complete healing through the canonical BMP/Smad signaling. Moreover, the IMC/AgNWs enhance vertical bone height and osseointegration in peri‐implantitis in beagle dogs, indicating the clinical translational potential of IMC/AgNWs for infectious vertical bone augmentation.

Purpose: To explore the esophageal cancer (EC) incidence and mortality trends and risk factors in China during 2005– 2015. Materials and Methods: The data were stratified by area (urban, rural), gender (male, female), and age groups (0 ∼, 5 ∼, …, 85 ∼). The age-standardized incidence rate (ASIR) and mortality rate (ASMR), age-specific incidence and mortality were calculated to describe the trends, which were analyzed by Joinpoint software, negative binomial regression model, and age-period-cohort model. Results: Trends in EC ASIR decreased markedly during 2010– 2015 (APC=− 6.14%, P< 0.05), and the average annual percent change (AAPC) value was − 8.07% (95% confidence interval (CI): − 9.98∼− 6.12) for rural areas during 2005– 2015. The ASMR was on a fast-downward trend after 2011 (APC=− 6.67%, P< 0.05), with AAPC values of − 1.34% (95% CI: − 2.56∼− 0.19) for males, − 3.39% (95% CI: − 5.65, − 1.07) for females, and − 9.67% (95% CI: − 10.56∼− 8.77) for rural areas during 2005– 2015. The age-specific incidence and mortality increased with age. The risk of EC for males was 3.1675 times higher than females (P< 0.001), and for urban areas, it was 0.58 times larger than rural (P< 0.001). The age and period effects presented an increasing trend, with a decreasing trend for the cohort effects in incidence and mortality risk. Later birth cohorts presented lower risks than previous birth cohorts. Conclusion: ASIR and ASMR in China are higher in males than females, and higher in rural than urban areas, which have decreased during 2005– 2015, especially in rural areas. The incidence increased with age up to the peak age group of 75. Area, gender, and age were independent risk factors for EC incidence. Keywords: esophageal cancer, junction regression, negative binomial regression model, age-period-cohort analyses

The kidney is important in the long-term regulation of blood pressure and sodium homeostasis. Stimulation of ETB receptors in the kidney increases sodium excretion, in part, by decreasing sodium transport in the medullary thick ascending limb of Henle and in collecting duct. However, the role of ETB receptor on Na(+)-K(+) ATPase activity in renal proximal tubule (RPT) cells is not well defined. The purpose of this study is to test the hypothesis that ETB receptor inhibits Na(+)-K(+) ATPase activity in rat RPT cells, and investigate the mechanism(s) by which such an action is produced. In RPT cells from Wistar-Kyoto rats, stimulation of ETB receptors by the ETB receptor agonist, BQ3020, decreased Na(+)-K(+) ATPase activity, determined by ATP hydrolysis (control=0.38+/-0.02, BQ3020=0.26+/-0.03, BQ788=0.40+/-0.06, BQ3020+BQ788=0.37+/-0.04, n=5, P<0.01). The ETB receptor-mediated inhibition of Na(+)-K(+) ATPase activity was dependent on an increase in intracellular calcium, because this effect was abrogated by a chelator of intracellular-free calcium (BAPTA-AM; 5 x 10(-3) M 15 min(-1)), Ca(2+) channel blocker (10(-6) M 15 min(-1) nicardipine) and PI3 kinase inhibitor (10(-7) M per wortmannin). An inositol 1,4,5-trisphosphate (IP3) receptor blocker (2-aminoethyl diphenyl borate; 10(-4) M 15 min(-1)) also blocked the inhibitory effect of the ETB receptor on Na(+)-K(+)ATPase activity (control=0.39+/-0.06, BQ3020=0.25+/-0.01, 2-APB=0.35+/-0.05, BQ3020+ 2-APB=0.35+/-0.06, n=4, P<0.01). The calcium channel agonist (BAY-K8644; 10(-6) M 15 min(-1)) inhibited Na(+)-K(+) ATPase activity, an effect that was blocked by a phosphatidylinositol-3 kinase inhibitor (10(-7) M 15 min(-1) wortmannin). In rat RPT cells, activation of the ETB receptor inhibits Na(+)-K(+) ATPase activity by facilitating extracellular Ca(2+) entry and Ca(2+) release from endoplasmic reticulum.

Although percutaneous posterior-ring tension-band metallic plate and percutaneous iliosacral screws are used to fix unstable posterior pelvic ring fractures, the biomechanical stability and compatibility of both internal fixation techniques for the treatment of Denis I, II and III type vertical sacral fractures remain unclear.Using CT and MR images of the second generation of Chinese Digitized Human "male No. 23", two groups of finite element models were developed for Denis I, II and III type vertical sacral fractures with ipsilateral superior and inferior pubic ramus fractures treated with either a percutaneous metallic plate or a percutaneous screw. Accordingly, two groups of clinical cases that were fixed using the above-mentioned two internal fixation techniques were retrospectively evaluated to compare postoperative effect and function. Parallel analysis was performed with a finite element model controlled trial and a case control study.The difference of the postoperative Majeed standards and outcome rates between two case groups was no statistically significant (P > 0.05). Accordingly, the high values of the maximum displacements/stresses of the plate-fixation model group approximated those of the screw-fixation model group. However, further simulation of Denis I, II and III type fractures in each group of models found that the biomechanics of the plate-fixation models became increasingly stable and compatible, whereas the biomechanics of the screw-fixation models maintained tiny fluctuations. When treating Denis III fractures, the biomechanical effects of the pelvic ring of the plate-fixation model were better than the screw-fixation model.Percutaneous plate and screw fixations are both appropriate for the treatment of Denis I and II type vertical sacral fractures; whereas percutaneous plate fixation appears be superior to percutaneous screw fixation for Denis III type vertical sacral fracture. Biomechanical evidence of finite element evaluations combined with clinical evidence will contribute to our ability to distinguish between indications that require plate or screw fixation for vertical sacral fractures.

Abstract The self-renewal ability is a unique property of fetal-derived innate-like B-1a lymphocytes, which survive and function without being replenished by bone marrow (BM) progenitors. However, the mechanism by which IgM-secreting mature B-1a lymphocytes self-renew is poorly understood. In this study, we showed that Bmi1 was critically involved in this process. Although Bmi1 is considered essential for lymphopoiesis, the number of mature conventional B cells was not altered when Bmi1 was deleted in the B cell lineage. In contrast, the number of peritoneal B-1a cells was significantly reduced. Peritoneal cell transfer assays revealed diminished self-renewal ability of Bmi1-deleted B-1a cells, which was restored by additional deletion of Ink4-Arf, the well-known target of Bmi1. Fetal liver cells with B cell–specific Bmi1 deletion failed to repopulate peritoneal B-1a cells, but not other B-2 lymphocytes after transplantation assays, suggesting that Bmi1 may be involved in the developmental process of B-1 progenitors to mature B-1a cells. Although Bmi1 deletion has also been shown to alter the microenvironment for hematopoietic stem cells, fat-associated lymphoid clusters, the reported niche for B-1a cells, were not impaired in Bmi1−/− mice. RNA expression profiling suggested lysine demethylase 5B (Kdm5b) as another possible target of Bmi1, which was elevated in Bmi1−/− B-1a cells in a stress setting and might repress B-1a cell proliferation. Our work has indicated that Bmi1 plays pivotal roles in self-renewal and maintenance of fetal-derived B-1a cells.

Objective To report outcomes of single-stage surgery under topical anaesthesia for the treatment of small-angle strabismus. Design Case series. Participants Thirteen patients, 7 males and 6 females, with a median age of 32 years (range, 20–59 years) were included. Methods Patients with symptomatic small-angle strabismus with stable deviations of no more than 20 prism diopters (PD) in horizontal and 10 PD in vertical were consecutively recruited from the Eye and ENT Hospital of Fudan University between January 2010 and April 2012. Single-stage surgery was performed under topical anaesthesia. Outcome measures were PD, Amblyopia and Strabismus Questionnaire (ASQE) scores, and subjective reduction of symptoms. Results The median duration of symptoms was 40 months (range, 6–96 months). Nine patients had horizontal deviations, 3 had vertical deviations, and 1 had an exodeviation combined with a vertical deviation. All surgeries were completed without complications, and no patients experienced significant discomfort. All patients reported elimination of symptoms on postoperative day 1. Two patients required a second procedure at 1 week because of a return of symptoms. At 6-month follow-up, no patient reported recurrence of symptoms. The overall ASQE score improved from 70 preoperatively to 96 postoperatively (p = 0.001). Conclusions These results suggest single-stage surgery under topical anaesthesia is an effective treatment for small-angle strabismus. A large, randomized, prospective study to confirm these findings is warranted.

Background: Immunotherapy based on cytokine-induced killer cells or combination of dendritic cells and cytokine-induced killer cells (CIK/DC-CIK) showed promising clinical outcomes for treating esophageal cancer (EC).However, the clinical benefit varies among previous studies.Therefore, it is necessary to systematically evaluate the curative efficacy and safety of CIK/ DC-CIK immunotherapy as an adjuvant therapy for conventional therapeutic strategies in the treatment of EC. Materials and methods: Clinical trials published before October 2016 and reporting CIK/ DC-CIK immunotherapy treatment responses or safety for EC were searched in Cochrane Library, EMBASE, PubMed, Wanfang and China National Knowledge Internet databases.Research quality and heterogeneity were evaluated before analysis, and pooled analyses were performed using random-or fixed-effect models.Results: This research covered 11 trials including 994 EC patients.Results of this metaanalysis indicated that compared with conventional therapy, the combination of conventional therapy with CIK/DC-CIK immunotherapy significantly prolonged the 1-year overall survival (OS) rate, overall response rate (ORR) and disease control rate (DCR) (1-year OS: P=0.0005; ORR and DCR: P,0.00001).Patients with combination therapy also showed significantly improved quality of life (QoL) (P=0.02).After CIK/DC-CIK immunotherapy, lymphocyte percentages of CD3 + and CD3 -CD56 + subsets (P,0.01) and cytokines levels of IFN-γ, -2, TNF-α and IL-12 (P,0.00001) were significantly increased, and the percentage of cluster of differentiation (CD)4 + CD25 + CD127 -subset was significantly decreased, whereas analysis of CD4 + , CD8 + , CD4 + /CD8 + and CD3 + CD56 + did not show significant difference (P.0.05). Conclusion:The combination of CIK/DC-CIK immunotherapy and conventional therapy is safe and markedly prolongs survival time, enhances immune function and improves the treatment efficacy for EC.

Surname distribution could offer substantial information regarding population migration. In this study, based on surname distribution, a novel index for measuring the migration intensity of a single region is proposed – the SRMI (Single-Regional Migration Intensity) – and is applied at the scale of prefectures in China. The prefectures with similar values of SRMI tend to adjoin each other on the administrative map, resulting in several distinct areas. The migration intensity is the highest in the northeastern, followed by the Central Plain and the Yangtze River Basin, and the lowest in the southern China with a larger ethnic minority population. The index of SRMI provides a new approach to study migration intensity in other countries and regions. The empirical results offer a fresh perspective of the historical migration characteristics in China.

Abstract Adult tendon stem/progenitor cells (TSPCs) are essential for tendon maintenance, regeneration, and repair, yet they become susceptible to senescence with age, impairing the self-healing capacity of tendons. In this study, we employ a recently developed deep-learning-based efficacy prediction system to screen potential stemness-promoting and senescence-inhibiting drugs from natural products using the transcriptional signatures of stemness. The top-ranked candidate, prim-O-glucosylcimifugin (POG), a saposhnikovia root extract, could ameliorate TPSC senescent phenotypes caused by long-term passage and natural aging in rats and humans, as well as restore the self-renewal and proliferative capacities and tenogenic potential of aged TSPCs. In vivo, the systematic administration of POG or the local delivery of POG nanoparticles functionally rescued endogenous tendon regeneration and repair in aged rats to levels similar to those of normal animals. Mechanistically, POG protects TSPCs against functional impairment during both passage-induced and natural aging by simultaneously suppressing nuclear factor-κB and decreasing mTOR signaling with the induction of autophagy. Thus, the strategy of pharmacological intervention with the deep learning-predicted compound POG could rejuvenate aged TSPCs and improve the regenerative capacity of aged tendons.

Extended thymectomy is indicated for patients with myasthenia gravis (MG) when drug-resistance or dependence is seen. We have employed a technique for subcostal thoracoscopic extended thymectomy (STET) on patients with MG.Clinical data of 15 eligible patients who underwent STET in our department from February 2015 to November 2015 by the same surgical team were retrospectively analyzed. The operation time, blood loss, duration of postoperative hospital stay, thoracic drainage periods were concerned.All the surgeries were finished successfully without conversion to sternotomy. Mean operation time was 157.53±40.31 min (range, 73-275 min). Mean blood loss was 56.33±7.07 mL (range, 10-200 mL). Mean pleural drainage volume in the first 24 hours was 72.67±17.68 mL (range, 0-250 mL). Mean postoperative thoracic drainage periods were 1.20±0.71 days (range, 0-3 days). Mean duration of postoperative hospital stay was 6.13±0.71 days (range, 3-22 days).This procedure showed satisfactory results for patients with MG. Moreover, the STET approach is more easily for surgeons to fully reveal the bilateral phrenic nerve and the upper thymic poles. We believe that STET is a satisfactory procedure for performing extended thymectomy in well selected patients.

Aplastic anemia (AA) is an immune-mediated disorder and mainly related to active destruction of hematopoietic cells by effector T lymphocytes. T helper (Th) 22 cells characterized as a novel subset of CD4+ T cells participate in the pathogenesis of autoimmune and hematological diseases. However, the role of Th22 subset in AA remains unknown.31 untreated AA patients and 30 healthy controls were included in this study. The percentages of Th22, Th17 and pure Th17 cells in peripheral blood were detected by flow cytometry. ELISA to measure interleukin (IL)-22 and IL-17A plasma levels and qRT-PCR for the mRNA levels of Th22 and Th17 related molecules were performed.The proportions of Th22, pure Th17, Th17 cells and plasma levels of IL-22 were significantly lower in untreated AA patients than those in normal controls. A positive correlation was found between Th22 and pure Th17 cells in AA. Moreover, percentages of Th22 cells correlated positively with reticulocyte counts and percentages. In addition, STAT3/STAT5 mRNA expression ratio was elevated in AA patients.Together, our results showed Th22 cells correlating with clinical characteristics of AA patients, indicating a possible role of Th22 immune response in the pathogenesis and therapeutic intervention of AA.

Cooperative behavior depends on cultural environment, so what happens when people move from to a new culture governed by a new norm? The dynamics of culture-induced cooperation has not been well understood. We expose lab participants to a sequence of different subject pools while playing a constrained Trust Game. We find prior exposure to different subject pools does in fact influence cooperative behavior; first impressions matter-the primacy effect plays a stronger role than the recency effect; and selfish first impressions matter more than cooperative first impressions-observing selfish behavior by others had a longer-lasting and greater influence on behaviors than observing cooperative behavior by others. Moreover, three consecutive exposures to cooperative environments were needed to neutralize one exposure to a selfish environment.

Abstract Objective We propose an index to characterize the key feature of Chinese surname distributions and investigate its implications for population structure and dynamics. Materials and methods The surname dataset was obtained from the National Citizen Identity Information Center, which contains 1.28 billion Chinese citizens enrolled in 2007, excluding those of Hong Kong, Macao, and Taiwan. An index, the coverage ratio of stretched exponential distribution (CRSED), is proposed based on the crossover point of stretched exponential truncated power‐law distribution, where the stretched exponential term and the power‐law term contribute equally. We use multidimensional scaling technique to demonstrate the dependence of the similarity of one prefecture to the others on the CRSED. Results The CRSEDs of 362 prefectures exhibit an uneven distribution. The consistency of this index is evident by strong positive correlations of CRSEDs at the three administrative levels. This new index has a strong negative correlation with the proportion of the rare surnames. The prefectures with similar CRSEDs tend to adjoin each other on the administrative map, resulting in several distinct regions, each of which shares similar terrain features or historical migrations. The prefectures with lower CRSEDs are more dissimilar to the other prefectures, while the ones with higher CRSEDs are more similar to the others. Discussion The population dynamics of the prefectures with higher CRSEDs are more likely dominated by migratory movements, the dominant evolutionary forces of the prefectures with lower CRSEDs can be attributed to drift and mutation.

Polyotia is a very rare auricular malformation, and only few cases have been reported to date. Polyotia has been ambiguously defined, and due to the instability of its shape and condition, no uniform surgical technique has been established up to now. Thus, it is necessary to standardize the diagnosis and treatment of polyotia. The aim of the present study was to present a new set of objective diagnostic criteria for discussion, and introduce our surgical design for polyotia.A retrospective analysis was performed on 34 cases of polyotia, which were diagnosed and treated in our Plastic Surgery Department during a 3-year period from January 2016 to March 2019. The preoperative photographs, manifestations and operation records of these 34 cases were reviewed.On the basis of the new set of objective diagnostic criteria, only 12 of 34 cases were diagnosed as polyotia, while the remaining 22 cases were diagnosed as accessory tragus. Polyotia was redefined as the presence of a broad-based accessory auricle in the tragus area along with accessory cavitas conchae similar to cavitas conchae. The new surgical design emphasized the use of cartilage and skin to fill up the concavity and reconstruct the tragus.The diagnosis of polyotia was presented on the basis of a new set of objective criteria, which include an accessory auricle and accessory cavitas conchae. The use of cartilage and skin to fill up the concavity and reconstruct the tragus were the emphases.

Objective To find out an effective therapy for autism. Methods Sixty children of autism were randomly divided into an electroacupuncture (EA) plus behavior therapy group and a behavior therapy group, 30 cases in each group. The patients in the two groups were treated with routine behavior, with EA at Baihui (GV 20), Sishencong (EX-HN 1), Shenting (GV 24), Benshen (GB 13), Yintang (EX-HN 3), Naohu (GV 17), Naokong (GB 19), Neiguan (PC 6) and scalp acupuncture at Speech Areas I, II, III added for the EA plus behavior therapy group. Their therapeutic effects were observed, and the picture and vocabulary scale (PPVT) and behavior ability were detected. Results The total effective rate was 86.7% in the EA plus behavior therapy group which was better than 56.7% of the behavior therapy group, and had significant enhancement in sensation, association, body, and ability of self-care (P 0.05). Conclusion EA combined with behavior therapy can significantly improve clinical symptoms of autism, but does not improve intelligence.

Abstract Exosomes are nanosized membrane‐bound extracellular vesicles with the capacity to modify functions and behaviors of recipient cells via transferring unique biological information. Mounting evidence suggests that under developmental conditions, mesenchymal cell‐derived exosomes are capable of mediating intercellular communication between epithelium and mesenchyme during organogenesis of salivary gland, tooth, hair follicle, and kidney, shedding light on potential therapeutic benefits elicited by mesenchymal exosomes in nanomedicine. Representing the most extensively investigated multipotent stem cells, mesenchymal stem cells (MSCs) have been identified and extracted from a wide variety of tissues. Accordingly, exosomes derived from MSCs (MSC‐Exos) can be easily accessed and manipulated as nanoparticles, giving rise to an entirely new therapeutic agents for nanomedicine. MSC‐Exos have presently emerged as efficacious alternatives to MSCs for cell‐free therapeutic strategies in disease contexts of ischemic heart disease, cutaneous wound healing and coronavirus disease 2019. In this review, we first present a critical analysis of research state of exosomes in terms of their biogenesis, molecular composition, isolation and characterization, then summarize current understanding of developmental regulation mediated by mesenchymal cell‐derived exosomes and reparative potential conferred by MSC‐Exos, along with particular emphasis on the underlying mechanisms of action and commentary on the major outstanding issues faced with MSC‐Exos.

Patching is often less effective for severe amblyopia because of poor adherence. For the treatment of severe amblyopia, although combined atropine and patching therapy (CAPT) has been found to be efficacious, it is currently unknown whether CAPT is more efficacious than patching alone.To compare the efficacy of CAPT vs patching alone in children aged 3 to 12 years with severe amblyopia.This single-center randomized clinical trial was conducted from November 2018 to May 2020. The visual acuity (VA) examiner was masked to the treatment groups. The follow-up visits were at 3 months and 6 months. Participants aged 3 to 12 years with severe amblyopia (20/100 to 20/500) resulting from strabismus, anisometropia, or both were randomly assigned to CAPT or patching therapy.CAPT or patching alone for 6 months.Change of the amblyopic eye VA from baseline to 6 months.Among 108 participants, the mean (SD) age was 5.2 (1.8) years, and 54 (50%) were female. Overall, 53 participants (49%) were randomized to CAPT and 55 (51%) were randomized to patching therapy. At baseline, the mean (SD) amblyopic eye VA was 0.95 (0.22) logMAR (approximately 20/200 [2.2 lines]). At 6 months, the CAPT group's mean improvement in amblyopic eye VA was 0.72 logMAR (7.2 lines) compared with 0.58 logMAR (5.8 lines) in the patching alone group (difference, 0.14 logMAR [1.4 lines] greater in the CAPT group; 95% CI, 0.05-0.22 logMAR [0.5-2.2 lines]; P = .002). The amblyopic eye VA improvement in the CAPT group also was greater than that in the patching alone group at 3 months (difference in the means, 0.13 logMAR [1.3 lines]; 95% CI, 0.04-0.22 logMAR [0.4-2.2 lines]; P = .004). No participants were withdrawn because of adverse effects.CAPT resulted in more mean improvement of amblyopic eye VA than patching alone among participants enrolled in this trial, although the clinical relevance of this relatively small VA difference cannot be determined from this trial.Chinese Clinical Trial Registry Identifier: ChiCTR1800018663.

Background: Disability weights (DWs) is basically pivotal parameter for disease burden calculation and to quantify the severity of health states from disease sequela. This study aimed to conduct a big survey and construct national and subnational DWs in mainland China and its provinces, and evaluate if the characteristics of age, sex, disease experience status, etc. have an impact on the valuation of these DWs.Methods: We conducted a web-based survey to assess DWs for 206 health states in 31 Chinese provinces in 2020-2021. We used two versions of DW questionnaire. The first version consisted of 16 paired comparison (PC) and 3 population health equivalence (PHE) questions; the second included 3 PC and 4 PHE questions. The health states that were depicted in the PC and PHE questions were randomly assigned to each respondent. The PC data was analyzed by probit regression analysis, and the regression results were anchored by results from the PHE responses on the DW scale units between 0 (no loss of health) and 1 (loss equivalent to death). The Pearson correlation analysis was performed for the probit coefficients between provinces and within province by participants’ characteristics.Findings: We considered 468 541 nationally representative respondents. The national DWs were bounded by mild distance vision impairment or mild anemia (0·009 [95% UI 0·0003-0·057]) and severe heroin and other opioid dependence (0·752, 0·640-0·841). In subnational analysis, we observed good PC responses and high correlations by province compared with the national data (p < 0·001): the lowest DW in distance vision mild impairment or mild anemia ranged from 0·008 (Henan and Ningxia) to 0·013 (Xinjiang) and the highest DW in severe heroin and other opioid dependence with a range of 0·693 (Henan)-0·813(Ningxia). Most of Chinese DWs for diabetes and digestive and genitourinary disease, mental, behavioural, and substance use disorders, hearing and vision loss, and disfigurement were larger than the GBD 2013 DW. The liner regression showed health states with mobility, mental and pain symptoms were significantly associated with lower DW in China compared with GBD 2013 and Japan; pain and sensory symptom with a higher Japanese DW than GBD 2013. Despite there are considerable disagreement, the DWs from the three regions are all highly correlated (p < 0·001). we put insights in other factors that might impact the valuation and found a slightly lower correlation of the probit coefficients between provinces (range rs: 0·980–0·997) than between medical background (rs = 0·985), profession (range rs = 0·987-0·998), income levels (range rs: 0·991–0·998), age groups (range rs: 0·992–0·998), educational level (rs: 0·991), sex (rs = 0·997) and disease status (rs = 0·998) (p< 0·001). Importantly, within province the lowest correlations of the probit coefficients were between low and high income level (range rs: 0·847–0·985, p < 0·001).Interpretation: This study created an empirical basis for national and subnational DW measurement in China. The considerable differences suggest that there might be contextual differences in evaluating the severity of health states between GBD regions and China, even among Asian countries. Apart from contextual differences, we found variations between income levels in health valuation within province, thus the effect of income level might be considered into valuating the severity of disease sequela.Funding Information: This work was supported by the National Key Research and Development Program of China [grant numbers 2018YFC1315302], the National Natural Science Foundation of China [grant number 81773552], and Wuhan Medical Research Program of Joint Fund of Hubei Health Committee [grant number WJ2019H304].Declaration of Interests: All other authors declare no competing interests.Ethics Approval Statement: This study was approved by the Ethics Committee of Medical Department of Wuhan University (2019YF2055), and a waiver of written informed consent obtained from participants prior to web-based survey participation was approved.

In Reply Dr Wang comments on the style of acupuncture, the lack of correlation between improvements in urinary leakage and urine pad use, and the quality control of acupuncture treatments in our study.

Various techniques have been performed for repairing transverse facial clefts. This study aimed to investigate an optimal method for repairing transverse facial clefts. Twenty-seven patients from 2008 to 2017 were evaluated. Their mean age at repair was 6.7 months with a follow-up period of 6 months to 10 years. A method using an inferior lip-based triangular mucosa flap and a superior lip-based rectangular vermilion-mucosa flap was designed for transposition. The orbicularis oris was reconstructed by using everting mattress suture. The skin was sutured using linear cutaneous closure with a single superiorly rotated Z-plasty lateral to the commissure. A postoperative symmetrical commissure was obtained owing to complete contraction with the new commissure directed 2 or 3 mm medial to the symmetrical point on the lips individually for the 27 patients. Lateral displacement of the reconstructed commissure was not observed. The patients showed a plump and symmetrical cheek on the cleft side. Twenty-one patients with hemifacial microsomia achieved a prominent improvement compared with their preoperative appearance, although the postoperative cheeks still did not show fullness because of the lesser facial tissue on the cleft side. In the early follow-up period, most patients showed a minimal scar during movement. However, the scar became thinner and symmetrical oral movement was achieved over time. This method obtained a natural oral movement without a conspicuous scar and was reliable and remarkable for the postoperative appearance of commissural symmetry. We conclude that this is an optimal method to repair transverse facial clefts.

BACKGROUND: Vaccination of dendritic cells (DCs) with tumor antigens is a potential strategy to induce tumor‐specific immunity in tumor‐bearing patients. The purpose of this study was to investigate whether human monocyte‐derived DCs were able to present P210 Bcr‐Ab1 protein and induce antigen‐specific cytotoxic T lymphocyte (CTL) responses in vitro after transfected with total RNA of K562 cells (K562‐RNA). STUDY DESIGN AND METHODS: DCs derived from human peripheral blood mononuclear cells were transfected with K562‐RNA with electroporation or DOTAP lipofection. The successful transfection was determined by reverse transcription‐polymerase chain reaction and Western blot. The phenotypes of the DCs were analyzed by flow cytometry (FCM), and cytotoxicity of CTL was assessed by propidium iodide staining followed by FCM analysis. The CD1a expression and purity of DCs were measured by FCM. RESULTS: The Bcr‐Abl fusion gene was detected in the DCs with 24 hours after the transfection. The transfected cell expressed increased levels of CD80, CD83, CD86, and HLA‐DR. Moreover, the transfected DCs strongly stimulated the T lymphocytes to gain cytotoxic activity against K562 cells. Culture medium containing 1 percent human plasma was the most effective for DC growth. CONCLUSION: Human DCs transfected with K562‐RNA effectively induce specific immune responses. This method can be used to induce tumor‐specific immune response and may have potential application in immunotherapy of tumors.

Objective: To analyze the subjective and objective results and the correlation of nasal obstruction because of nasal structural variation pre and post operation, and to provide clinical evidence for surgery and curative effect. Methods: Three groups were included in this study: group 1 were 78 patients with structural rhinitis, group 2 were 72 patients with chronic sinusitis (without nasal polyp) and group 3 were healthy controls of 75 cases. Assessment of nasal obstruction was achieved by visual analog scale (VAS), and objective measurement was achieved by rhinomanometry and acoustic rhinometry. The first two minimum cross-sectional area of nasal cavity (MCA1, MCA2), their distances from the nostrils (MD1, MD2) and the nasal volume of 5 cm depth from the nostril (NV5) were recorded. The ratio of the parameters of the two sides of the nasal cavity was calculated respectively. Nasal resistance total (RT) and calculated right-left nasal resistance ratio (Rlr) were recorded. Patients of group 1 and group 2 accepted endoscopic surgery and received the above evaluation again at three months after surgery. Three groups were compared with statistical test. Results: The preoperative values of three groups (MCA1, MD1, MCA2, MD2, NV5, RT) showed no statistical significance (F value was 0.945, 0.245, 1.380, 0.036, 0.866, 1.651, respectively; all P>0.05), while their ratio had statistical differences except MD1 (F value was 5.242, 1.726, 4.882, 4.005, 5.066, 5.316; P=0.013, 0.199, 0.019, 0.024, 0.018, 0.011, respectively). Statistical results between the ratio of values (MCA1, MD1, MCA2, MD2, NV5, RT) and VAS of nasal obstruction of group 1 and group 2 showed significant positive correlations pre operation(r of group 1 value was 0.471, 0.418, 0.260, 0.324, 0.305, 0.459, respectively; r of group 2 value was 0.373, 0.403, 0.288, 0.366, 0.402, 0.249, respectively; all P<0.05). VAS scores pre and post operation of group 1 had statistically difference (t=35.122, P<0.05). Postoperative value of MD1, MD2 and RT among three groups had no statistical difference (F value was 0.178, 0.582, 0.905, respectively; all P>0.05). Postoperative value of MCA1, MCA2, NV5 among three groups had statistical difference (F value was 4.010, 5.126, 4.901, respectively; all P<0.05). Postoperative ratio of MCA1, MD1, NV5 and Rlr among three groups had no statistical difference (F value was 1.023, 0.944, 0.524, 0.996, respectively; all P>0.05). Postoperative ratio of MCA2 and MD2 among three groups had statistical difference (F value was 4.859, 4.357, respectively; all P<0.05). Conclusion: Severity of nasal obstruction is related to structural variation of nasal cavity and the purpose of surgery is to restore structural symmetry of bilateral nasal cavity.

We have developed an optimized model for electron behavior in Cu-line and we have implemented it using Monte Carlo method. Our model takes into account not only four normal scatterings but also the grain boundary scattering and the surface roughness scattering. The model has been tested with different line width and providing a good agreement with both calculated results and ITRS data.

Patients undergone septoplasty, which is one of the most common procedures, always feel very painful after surgery because of routine nasal packing. The objective of this study was to evaluate the effect of septal suture technique without nasal packing after septoplasty.Eighty patients who had undergone septoplasty were included in this study. The patients were allocated into tow groups: packing group, mercel was used for nasal packing after septoplasty and inferior turbinate coblation; suturing group, septal suture and inferior turbinate coblation were performed after septoplasty without nasal packing. Postoperative signs and symptoms(visual analogue scale,VAS) were compared between the two groups.The patients of packing group experienced significantly more postoperative nasal pain, headache, dysphagia, sleep disturbance and bleeding after surgery (P < 0.01). No difference of epiphora was found between the two groups. More pain and bleeding were experienced during the pack removal(packing group), compared to that during the clearance of nasal cavity (suturing group ). One patient with postoperative septal hematoma was found in the packing group.Nasal septum suture combined with inferior turbinate coblation might be a significantly more comfortable, reliable alternative to nasal packing. Nasal packing is not the necessary application for septoplasty.

Abstract Recruiting endogenous stem cells to bone defects without stem cell transplantation and exogenous factor delivery represents a promising strategy for bone regeneration. Herein, we develop an alkaline shear-thinning micro-nanocomposite hydrogel (10-MmN), aiming to alkaline-activate endogenous TGFβ1 and achieve in situ bone regeneration. It contains polyethyleneimine (PEI)-modified gelatin, laponite nanoplatelets (LAP), a bicarbonate buffer with a pH of 10, and gelatin microspheres (MSs). PEI-modified gelatin plays a pivotal role in hydrogel fabrication. It endows the system with sufficient positive charges, and forms a shear-thinning nanocomposite matrix in the pH 10 buffer (10-mN) with negatively charged LAP via electrostatic gelation. For biological functions, the pH 10 buffer dominates alkaline activation of endogenous serum TGFβ1 to recruit rat bone marrow stem cells through the Smad pathway, followed by improved osteogenic differentiation. In addition, MSs are incorporated into 10-mN to form 10-MmN, and function as substrates to provide good attachment sites for the recruited stem cells and facilitate further their osteogenic differentiation. In a rat critical-sized calvarial defect model, 10-MmN exhibits excellent biocompatibility, biodegradability, hydrogel infusion and retention in bone defects with flexible shapes and active bleeding. Importantly, it repairs ~95% of the defect areas in 3 months by recruiting TGFβR2 + and CD90 + CD146 + stem cells, and promoting cell proliferation, osteogenic differentiation and bone formation. The present study provides a biomaterial-based strategy to regulate alkalinity in bone defects for the initiation of endogenous TGFβ signaling, which can be extended to treat other diseases.

Abstract Numerous cells grow in columnar tissues and organs with different curvatures and curvature gradients. Therefore, it is necessary to study the effect of curvature on cell behavior to control and promote cell development. Herein, we prepared polydimethylsiloxane (PDMS) with different micro‐nano patterns using ultraviolet soft lithography. Hydrophilic polydopamine (PDA) was modified on the PDMS surface to prepare PDMS/PDA to improve its biocompatibility. The PDMS/PDA was characterized by contact angle tester and scanning electron microscopy (SEM). The effect of curvature on bone cell migration and differentiation was studied through SEM, inverted phase contrast microscope and fluorescence microscopy. We found that different curvatures had different effects on the bone cell migration and differentiation. Chondrocytes migrated rapidly in grooves with a curvature range of 1/575–1/875 μm −1 . Bone mesenchymal stem cells (BMSCs) had high efficiency of differentiation into chondrocytes in the grooves with a curvature range of 1/775–1/1375 μm −1 . Furthermore, BMSCs showed high efficiency of differentiation into chondrocytes at the edges of micro‐nano patterns with different perimeter curvatures, and the differentiation efficiency was the highest at 120° convex curvature. This work shows that curvature is a principle to be considered in bone tissue regeneration engineering and provides inspiration for future biomaterials design.

Whether anesthesia methods affect malignant biological behavior of cancer remains unresolved. In this study, we aim to compare the effects of general anesthesia (GA) and local anesthesia (LA) on serum collected from primary hepatocellular carcinoma (HCC) patients presenting for radiofrequency ablation (RFA).From August 2020 to December 2020, a prospective, randomized, and controlled study was conducted at Renji Hospital, which is affiliated with Shanghai Jiaotong University School of Medicine. 25 qualified patients from 18 to 65 years of age undergoing RFA were enrolled in the study and randomly assigned into two groups: the GA group (n = 14) and the LA group (n = 11). Venous blood was drawn from all patients preoperatively and 1 hour postoperatively. The serum collected was then used for the culturing of HepG2 cells. The malignant biological behaviors of HepG2 cells, including invasion, migration and proliferation, were observed after 24 hours of exposure to patients' serum. ELISA was used to compare expression levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and lymphokines (IFN-γ, IL-2) in patients' serum from both groups.HepG2 cells cultured with postoperative serum obtained from patients who received GA, but not LA, were associated with significantly increased cell invasion, migration and proliferation, compared to preoperative serum from the same patient group. Expression levels of pro-inflammatory cytokines were significantly higher, and lymphokines significantly lower in postoperative serum from GA patients compared to the corresponding preoperative serum.GA affects the serum milieu of patients with HCC, promoting the malignant biological behavior of a human HCC cell line.

To analyze the therapeutic effect of plate cable system and cortical strut bone graft in the prosthesis revision of the total hip arthroplasty for the treatment of Vancouver type B1 periprosthetic femoral fracture.A total of 8 patients were selected from January 2006 to January 2013, including 6 males and 2 females, aged from 56 to 74 years old (62.52 years old on average). All the cases were treated by the long plate cable system and appropriate cortical strut bone graft. Harris score was used to evaluate the hip functions before and after the operation. Prosthesis stability and the bony union were assessed by the digital radiography.All the patients were followed up for 45 months on average ranging from 24 to 60 months after operation. All the fractures reached union, and there was no infection, stem loosening, nonunion and malunion. The Harris score was 28.45±5.78 before operation, which was improved to 83.46±10.21 after operation. X ray showed that the prosthesis was stable, and the host bone and bone graft achieved bony union in 7 patients;and the other 1 patient need further operation of revision around the loose stem.The prosthesis revision of the total hip arthroplasty with the locking plate and cortical strut bone graft used for the Vancouver type B1 periprosthetic proximal femur fractures has the advantages of simple manipulation, less complications, good recovery of the hip function and can improve bone quality to provide favorable conditions for operation of revision.

A considerable body of researches indicate a high trend in researching social media. Accordingly, the study aimed to explore the relationship between social media relationships and academic self-concept through a correlation analysis. The data was collected from the 246 participants in secondary school who aged 12-16 years old from Shanghai, China. Results showed that all measures constructed for the study were internally consistent. Giving clues about social media, this study reveals that most students used blog, QQ, and WeChat frequently to connect with each other or share posts. The findings show there is a significant high positive relationship among positive peer relationships, academic persistence, academic expectation, learning interest, and academic self-concept.

Lentivector immunization stimulates potent CD8 T cell responses against melanoma self-antigen tyrosinase-related protein 1 and generates antitumor immunity in mice. [ J. Immunol. 182: 5960–5969][1] . The fifth institution in the affiliation line is incorrect. The corrected list is shown below. *

To establish a method for isolating exosomes from dendritic cells (DC), and to analyse its biological characteristics and function in antitumor immunity.Immature DCs (im-DC) from human peripheral blood mononuclear cells were loaded with the antigen of K562 tumor cells, then exosomes were secreted from imDC and lipopolysaccharide (LPS) induced mature DC (mDC). The exosomes from imDC and mDC were isolated separately by ultracentrifugation and ultrafiltration. The exosomes diameter was determined, their profile was observed by electron microscope, and the surface molecules were detected by Western blot. To analyse the effect of exosomes on antitumor immunity, the proliferation, IFN-gamma expression, CD69 up-regulation and cytotoxicity of antigen-specific T cells were measured.Exosomes were small flattened sphere vesicles with an average diameter of 72.3 nm and expressed CD80, CD86, HLA-DR, FasL, CD54 and MFG-E8 molecules. As compared to immature exosomes, exosomes from mDC were proved to express more CD80 and less MFG-E8, to be more potent for inducing antigen-specific T cells proliferation and immunity respond in vitro: at its optimum concentration, the absorption value of T cell proliferation test was 0.50 +/- 0.01, CD69 was up-regulated and (13.4 +/- 5.8)% of T cells was in proliferating, (22.8 +/-2.4)% of T cells expressed IFN-gamma, and (21.3 +/-8.6)% of tumor cells were killed.A simple and quick method to isolate and analyse exosomes is established. The exosomes can induce antitumor immunity respond.

目的 探讨使用交锁髓内钉治疗股骨干粉碎性骨折的手术方式选择、疗效.方法对1998年1月～2002年10月使用交锁髓内钉治疗67例(76肢)股骨干粉碎性骨折的疗效进行回顾性分析.手术采用闭合、小切口或有限切开复位,用三维瞄准器锁定骨折远近端.结果平均随访14.1个月,肢体、关节功能恢复优良率92.5%(62/67),一期骨折愈合率90.8%(69/76).平均骨临床愈合时间6.7个月.2例软组织感染,感染率2.6%,2例髓内钉断裂.结论交锁髓内钉治疗股骨干粉碎性骨折应常规静力固定,不剥离或少剥离骨膜,不强求解剖复位,尽量闭合复位、有限扩髓,是目前治疗股骨干粉碎性或多节段骨折的有效方法。

Objective: To introduce an improved direct method for first puncture and to study the safety of first puncture in laparoscopic surgery.Methods: Four kinds of techniques were used to perform the first puncture on 4 102 patients.Group A: trocar puncture were performed after pneumoperitoneum in 2 492 cases.Group B: trocar puncture was directly through the incision of umbilical skin in 798 patients; Group C: trocar puncture was directly through the incision of umbilic skin,subcutancous fat and fascia in 387 patients.Group D: opened umbilical hole and inserted the trocar shell without puncture in 425 cases.it was a success if entered the abdominal cavity at the first time,it was unsafe if punctured twice and a failure if the needle hadn't entered the abdominal cavity for 3 times or went into the subcutaneous vessels or viscera.Results: There were no vessels and viscera injury during punctures in our study.The puncture results were related to the different methods.Compared with group A, the success rate of group C and D were significantly improved( P 0.01).Group B was similar to that of group A( P 0.05).Success rates of group C and D were significantly higher than that of group A.The success rate was higher in group D than in group C( P 0.01).For 633 cases who received puncture by 38 learners,the results of punctrue were similar.There was no unsafety and failure case in group D.Conclusion: Direct trocar puncture after incision of umblical hole is safer and easier to perform than routine method.Trocar shell insertion is better than other methods especially for learning doctors.

The fetal liver is a major site containing progenitor cells to give rise to B-1 cells, especially CD5+ B-1a cells. We previously reported that hemogenic endothelial cells in the yolk sac (YS) and para-aortic splanchnopleure (P-Sp) (prior to hematopoietic stem cell [HSC] emergence) produced B-1 progenitors in vitro that engraft in immunodeficient neonates. Since the fetal liver is not a de novo site of hematopoietic cell emergence and must be seeded by various hematopoietic progenitor/stem cells derived from other hematopoietic tissues, B-1 progenitors in the fetal liver may be a mixture of YS/P-Sp derived progenies, and not only derivatives of HSCs that also seed the fetal liver at E11. Until now, it has been impossible to determine whether HSC independent B-1 progenitor cells exist in the fetal liver. We have utilized embryos genetically engineered to be devoid of HSC (CBFb-/-) but carrying a transgene expressing CBFb under the TEK promoter (CBFb-/-:TEK-CBFb/GFP mice). We have found the presence of transplantable B-1 and marginal zone B progenitors in the fetal liver despite the absence of HSCs. YS/P-Sp cells from CBFb-/-:TEK-CBFb/GFP embryos could produce B-1 progenitors in vitro. While TEK is expressed in hemogenic endothelial cells, it is not expressed in B-1 progenitors, and transplanted CBFb-/-:TEK-CBFb/GFP fetal liver cells failed to expand/maintain B-1 cells in the host peritoneum, unless CBFb expression was rescued by retrovirus transduction.Our data provide solid evidence for the presence of the first wave of HSC independent B-1 progenitors in the fetal liver as postulated 20 years ago in the immune layered theory of B lymphopoiesis. The fetal liver is a major site containing progenitor cells to give rise to B-1 cells, especially CD5+ B-1a cells. We previously reported that hemogenic endothelial cells in the yolk sac (YS) and para-aortic splanchnopleure (P-Sp) (prior to hematopoietic stem cell [HSC] emergence) produced B-1 progenitors in vitro that engraft in immunodeficient neonates. Since the fetal liver is not a de novo site of hematopoietic cell emergence and must be seeded by various hematopoietic progenitor/stem cells derived from other hematopoietic tissues, B-1 progenitors in the fetal liver may be a mixture of YS/P-Sp derived progenies, and not only derivatives of HSCs that also seed the fetal liver at E11. Until now, it has been impossible to determine whether HSC independent B-1 progenitor cells exist in the fetal liver. We have utilized embryos genetically engineered to be devoid of HSC (CBFb-/-) but carrying a transgene expressing CBFb under the TEK promoter (CBFb-/-:TEK-CBFb/GFP mice). We have found the presence of transplantable B-1 and marginal zone B progenitors in the fetal liver despite the absence of HSCs. YS/P-Sp cells from CBFb-/-:TEK-CBFb/GFP embryos could produce B-1 progenitors in vitro. While TEK is expressed in hemogenic endothelial cells, it is not expressed in B-1 progenitors, and transplanted CBFb-/-:TEK-CBFb/GFP fetal liver cells failed to expand/maintain B-1 cells in the host peritoneum, unless CBFb expression was rescued by retrovirus transduction. Our data provide solid evidence for the presence of the first wave of HSC independent B-1 progenitors in the fetal liver as postulated 20 years ago in the immune layered theory of B lymphopoiesis.

Objective To analyze the influence of different dosage of penehyclidine hydrochloride on postoperative cognitive function.Methods A total of 200 patients receiving operation treatment were enrolled and randomly divided into A,B,C,D and E group(200cases for each group),receiving injection of penehyclidine hydrochloride at dosage of 0.020,0.017,0.014,0.011 and 0.008mg/kg.Preoperative and tree-day postoperative Mini Mental State Examination(MMSE)scores,incidence rate of postoperative cognitive dysfunction and delirium,and thirst condition after injection were compared.Results Three-day postoperative MMSE scores decreased in the order of E,D,C,B and A group.,and in C,D and E group,the differences of MMSE scores detected preoperatively and three-day postoperatively were not significant(P0.05).Incidence rates of postoperative cognitive dysfunction and delirium decreased in the order of A,B,C,D and E group.10,20 and 30min after injection of penehyclidine hydrochloride,xerostomia score decreased in the order of A,B,C,D and E group,and with prolongation of time,the degree of xerostomia gradually increased in each group.Conclusion Application of penehyclidine hydrochloride at dosage of a certain range could reduce the incidence rate of postoperative cognitive dysfunction.

[Objective]To achieve activated OsVDAC3 protein in vitro.[Method]The recombinant prokaryotic expression vector pGEX-4T-1-osvdac3 with the GST-tag was constructed and transformed into E. coli. TheGst-tagged OsVDAC3 protein was induced with IPTG in E. coli strain BL21,and confirmed by SDS-PAGE and Western-Blot analysis. In addition,the GST-tagged recombinant OsVDAC3 protein was purified by Glutathione resin.[Result] The results showed that the recombinant prokaryotic expression vector pGEX-4T-1-osvdac3 was constructed and transformed into E. coli successfully. The results of SDS-PAGE and Western Blot showed a band in the 56 kD and indicated the GST-tag OSVDAC3 protein was partly existed in the soluble composition.[Conclusion]The GST-tag OsVDAC3 protein was isolated and purified by using Glutathione Resin. The research laid a foundation for the further study of the function of VDAC in the rice.

Objective To investigate the clinical efficacy of procedure for prolapse and hemorrhoids(PPH)to treat circumferential mixed hemorrhoid.Methods 86 patients were Collected with circumferential mixed hemorrhoid.According to the surgical method,43 patients received the PPH(PPH group)while the other 43 patients received the traditional Milligan surgery(control group).Patients in each group would be followed up with at least 6months after the surgery,and analyzed with the index with operation time,intraoperative force and discomfort,post-operation hospitalization duration,post-operation pain,post-operation bleeding,post-operation edema,post-operation skin tag occurance.Results Compared with the traditional Milligan surgery,patients received the PPH have the characteristic of shorter operation time,more intraoperative force and discomfort feeling,shorter post-operation hospitalization duration,less post-operation pain(P 0.05).The index difference of bleeding,skin tag occurance after the surgery all have no statistical significance(P 0.05).Conclusion PPH in the treatment of circumferential mixed hemorrhoid might be an alternative surgical method,the problem of more intraoperative force and discomfort feeling also should be concerned.

Objective To obtain highly-metastatic cells of squamous-cell-carcinoma of tongue and establish a base for its animal model. Methods Cells with different metastatic potential were isolated using modified invasion assay;clone differences of two kinds of cells were detected using MTT assay and cloning formation assay;the differences of cell invasiveness and migration ability by invasion assay and migration assay,that of cell cycle time by flow cytometry,and that of the enhancer of Zeste homolog 2(EZH2)protein and mRNA expression by Western Blot and Real-time PCR. Results The high and low metastatic potential cells(Rca-TH,Rca-TL)were isolated.The differences of cell colony formation,migration,invasion,cell cycle,EZH2protein and mRNA expression between Rca-TH and Rca-TL were statistically significant(t=3.423-59.300;P0.05,0.01). Conclusion Two different metastatic potential cell subclones were isolated from cell line of squamous cell cancer of tongue,and the biological characteristics between the Rca-TH and Rca-TL were significantly different.

To explore the clinical effect of using dorsal two wing-shaped flap to reconstruct finger web for treatment of congenital syndactyly.This technique has been used in 19 children with congenital syndactyly. At the dorsum, a flap with V-shaped tip and two wing-shaped pedicle were designed and was just sewed up with an anchor-shaped incision at the palm. The web was primarily reconstructed without skin graft at base of fingers. Distal end of fingers were separated by using serrated flap and were closed after removal of fatty tissue. At some cases with tight skin connection. The defect area at lateral and distal end of fingers was closed by small pieces of skin graft.All the webs were reconstructed primarily without skin graft at the base of fingers. 7 cases with tight skin connection had small pieces of skin graft at lateral and distal end of fingers. Primary healing was achieved in all cases. After 1 to 6 months of follow-up, both the appearance and function were satisfactory without conspicuous scar. The reconstructed finger webs were in normal depth and width.Primary web space can be achieved by dorsal two wing-shaped flap without skin graft at base of fingers. It is one of the best choices for treatment of congenital syndactyly.

The blind study design, particularly the double-blind study design is a very important method for diminishing placebo effect and reducing bias in clinical medical trial. Enlightened by Streitberger's and Park's sham needle design, the authors of the present paper introduce a newly designed sham needle device (Yan's sham-needle) for controlled double-blind trials of acupuncture. This sham needle device consists of needle, tube and base. The bottom of the tube is completely sealed and it can never arouse any invasive stimulation on the subject's skin when the sham needle is downward pressed on the body surface. Meanwhile, this sham device is filled with sponge which is able to simulate soft tissues of the acupoint area. By combining words suggestions or hints before trials and the same shape as verum device, this sham-needle device reduces the risk of blind-breaking and makes it possible to conduct controlled double-blind trials. Primary practice showed that this device may provide a new and practical tool for researching the placebo effect of acupuncture therapy.

Hyperactivation of the mTOR pathway impairs hematopoietic stem cell (HSC) functions and promotes leukemogenesis. mTORC1 and mTORC2 differentially control normal and leukemic stem cell functions. mTORC1 regulates p70 ribosomal protein S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E-binding (eIF4E-binding) protein 1 (4E-BP1), and mTORC2 modulates AKT activation. Given the extensive crosstalk that occurs between mTORC1 and mTORC2 signaling pathways, we assessed the role of the mTORC1 substrate S6K1 in the regulation of both normal HSC functions and in leukemogenesis driven by the mixed lineage leukemia (MLL) fusion oncogene MLL-AF9. We demonstrated that S6K1 deficiency impairs self-renewal of murine HSCs by reducing p21 expression. Loss of S6K1 also improved survival in mice transplanted with MLL-AF9-positive leukemic stem cells by modulating AKT and 4E-BP1 phosphorylation. Taken together, these results suggest that S6K1 acts through multiple targets of the mTOR pathway to promote self-renewal and leukemia progression. Given the recent interest in S6K1 as a potential therapeutic target in cancer, our results further support targeting this molecule as a potential strategy for treatment of myeloid malignancies.

Abstract The mixed-lineage leukemia (MLL) gene is required for the maintenance of adult hematopoietic stem cells (HSCs) and regulation of progenitor population. Translocations in MLL have been detected in approximately 5-10% of adult acute leukemia patients and in approximately 70% of acute leukemias in infants. Various genes, including AF4, AF5, AF9, ELL and ENL have been identified as partners for translocation in MLL-rearranged leukemia. In hematopoietic cells, expression of MLL fusion proteins result in a block of differentiation. AML patients who successfully undergo treatment but relapse suggest the importance of targeting leukemia initiating cells (LICs) within the MLL leukemia. LICs remain in a quiescent state and are capable of survival despite treatment with chemotherapeutic agents or targeted molecular inhibitors. In mouse models, LICs are defined by the capability of successfully propagating disease upon serial transplantation. In order to prevent disease relapse, identification of molecules, which regulate the self-renewal of LICs, is essential. The phosphatidylinositol 3-kinase (PI3K)-Akt-mechanistic target of rapamycin complex1 (mTORC1) pathway is a key regulator of self-renewal of both HSCs and LICs. Deletion of Raptor, a subunit of mTORC1, does not affect initiation and progression of acute myeloid leukemia (AML), but Raptor deficiency results in delayed propagation of AML. Upon its activation, mTORC1 phosphorylates and activates p70 ribosomal protein S6 kinase (S6K1) and inhibits the activity of eukaryote translation initiation factor 4E binding protein 1 (4E-BP1). S6K1 has been shown to be hyperactivated in hematopoietic cells expressing oncogenic MLL-AF9 fusion protein. In our study, we have assessed the role of S6K1 in the initiation, progression and propagation of AML using a genetic model of S6K1 knockout mice (S6K1-/-). We expressed MLL-AF9 fusion oncoprotein in WT and S6K1-/- hematopoietic stem and progenitor cells (HSC/Ps) and transplanted them into lethally irradiated recipients. Recipients of both WT and S6K1-/- HSC/Ps bearing MLL-AF9 displayed high white blood cell (WBC) count, splenomegaly and developed AML. There was no difference in survival between the WT and S6K1-/- recipients. In order to determine whether S6K1 regulates the self-renewal of LICs, we transplanted lethally irradiated mice with cells from WT and S6K1-/- primary recipients who developed AML. Recipients of S6K1 deficient AML cells survived significantly longer compared to controls (n=17/group, p&lt;0.001). S6K1 deficient HSC/Ps expressing MLL-AF9 showed reduced activation of Akt as well as decreased mTORC1 activity, suggesting that deletion of S6K1 results in reduced activation of PI-3K-Akt-mTORC1 pathway both upstream and downstream of mTORC1 which indicates that S6K1 might be involve in a feedback loop within this pathway. To determine the role of S6K1 in normal HSC development and maintenance, we analyzed bone marrow derived HSCs in WT and S6K1-/- mice. S6K1 deficiency did not alter the frequency of long term HSCs (LT-HSCs) as defined by CD150+ CD48- Lin- Sca1+ c-Kit+ surface markers, but the absolute number of LT-HSCs were significantly reduced in S6K1 deficient mice (p&lt;0.02). The absolute number of multipotent progenitors (MPPs) (p&lt;0.001), common myeloid progenitors (CMPs) (p&lt;0.01) and megakaryocyte-erythroid progenitors (MEPs) (p&lt;0.01) were also significantly reduced in S6K1 deficient mice. Deficiency of S6K1 resulted in reduced quiescence of LT-HSCs (p&lt;0.05). Expression level of p21, an inhibitor of cell cycle progression, was significantly decreased in LT-HSCs derived from S6K1-/- mice compared to LT-HSCs from control group. To study the role of S6K1 in HSCs' function, we performed competitive repopulation assay. Sorted LT-HSCs from bone marrow cells derived from either WT or S6K1-/- mice were transplanted with competitor cells into lethally irradiated recipients. S6K1 deficient LT-HSCs displayed reduced repopulating ability in secondary recipients (n=9-10/group, p&lt;0.001). Expression level of p21 was downregulated in donor-derived HSCs isolated from secondary recipients of S6K1 deficient HSCs compared to control. Overall, our study establishes S6K1 as a critical regulator of self-renewal of both LICs and HSCs. Deficiency of S6K1 in AML cells results in delayed propagation of disease and deficiency of S6K1 in HSCs results in decreased self-renewal potential. Disclosures No relevant conflicts of interest to declare.

Abstract Acute myeloid leukemia (AML) is a heterogeneous disease with multiple signaling pathways contributing to its pathogenesis. Mutations in receptor tyrosine kinase KIT and FLT3 are found in approximately 40% of AML patients and targeted therapies for inhibiting KIT and FLT3 have failed, thus new targets for therapeutic intervention need to be identified. The phosphatase of regenerating liver (PRL) family of phosphatases, consisting of PRL1, PRL2, and PRL3, represents an intriguing group of proteins being validated as biomarkers and therapeutic targets in human cancer. While PRL2 is highly expressed in some subtypes of human AML, including AML1-ETO+ AML and AML with mixed lineage leukemia (MLL) translocations, its role in AML is largely unknown. To determine the role of PRL2 in the pathogenesis of AML, we utilized two murine models of human AML induced by transducing mouse HSCs with AML1-ETO or MLL-AF9. We found that PRL2 is important for the progression and maintenance of leukemia induced by AML1-ETO or MLL-AF9 through enhancing leukemia stem cell (LSC) self-renewal. To elucidate the mechanisms by which PRL2 promotes LSC maintenance, we performed genome wide RNA-seq analysis of MLL-AF9+ LSCs. Gene Set Enrichment Analysis (GESA) indicates that PRL2 deficiency alters the MLL-AF9 signature essential for LSC self-renewal. We have recently identified PRL2 to be important for the proliferation and self-renewal of hematopoietic stem cells (HSCs) through the regulation of KIT signaling. Notably, PRL2 null hematopoietic progenitor cells showed decreased KIT phosphorylation as well as ERK phosphorylation following SCF stimulation, suggesting that PRL2 is important for KIT activation. Given that KIT inactivation could be mediated by removal from the cell surface and intracellular degradation, we reasoned that PRL2 may regulate KIT receptor internalization and stability. That was indeed the case. We found that the KIT protein half-life in PRL2 null hematopoietic progenitor cells (Kit+) was significantly decreased compared to WT cells. Furthermore, PRL2 null progenitor cells showed enhanced KIT ubiquitination compared to WT cells and less KIT was found on the surface of PRL2 null progenitor cells compared to WT cells following SCF stimulation. We also found that loss of PRL2 in human AML cells resulted in enhanced internalization of KIT. These observations demonstrate that PRL2 deficiency results in less KIT on the cell surface and a lower global KIT level in the cell. Upon SCF stimulation, KIT binds to and induces the phosphorylation of CBL proteins, which in turn act as E3 ligases, mediating the ubiquitination and degradation of KIT. To understand how PRL2 modulates the turnover of KIT in hematopoietic cells, we performed GST-pulldown assays and found that the substrate-trapping mutant PRL2/CS-DA showed an increased association with KIT and CBL compared to wild-type PRL2 in Kasumi-1 cells, suggesting that KIT and CBL may be PRL2 substrates. Furthermore, we found that PRL2/CS-DA mutant showed enhanced association with FLT3 and CBL compared to wild-type PRL2 in MV4-11 cells. Our data suggest that PRL2 dephosphorylates CBL and inhibits CBL activity toward KIT and FLT3, leading to sustained activation of downstream signaling pathways. To determine the functional significance of PRL2 in human AML with KIT and FLT3 mutations, we utilized two well-established murine model of myeloproliferative neoplasms (MPN) induced by KITD814V or FLT3-ITD. We found that loss of Prl2 decreased the ability of oncogenic KITD814V and FLT3-ITD to promote mouse hematopoietic stem and progenitor cell (HSPC) proliferation in vitro andthe development of MPN in vivo. Furthermore, we found that genetic and pharmacological inhibition of PRL2 decreased the proliferation and survival of human AML cells bearing KIT or FLT3 mutations. Taken together, we demonstrate that PRL2 promotes leukemia stem cell (LSC) self-renewal and maintenance through sustaining the activity of oncogenic KIT and FLT3 signals. Our findings suggest that pharmacological inhibition of PRL2 holds potential as a novel therapy for acute myeloid leukemia, and might also be applicable to the treatment of other human cancers. Disclosures No relevant conflicts of interest to declare.