Zhaoliang Su

This article presents a model-free predictive current control (MFPCC) method for dual three-phase permanent magnet synchronous machines with an optimal modulation pattern. First, an ultralocal model based on the <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">dq</i> subspace is established, and a novel nonlinear disturbance observer is designed to effectively compensate for voltage distortion caused by inverter characteristics. Additionally, the introduction of an intermediate variable eliminates the estimation peaks in the high-frequency state, improving both steady-state and transient performance of the controller while suppressing the influence of parameter mismatch on control performance. Second, two candidate modulation patterns and their corresponding pulse synthesis patterns are predefined, and the closed-loop control of the <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">xy</i> subspace is realized through an efficient cost function. The switching mode with the best <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">xy</i> current performance at the current moment is then switched in real time to obtain improved steady-state performance and deeper harmonic current suppression. Finally, the experimental results have verified the effectiveness of the proposed MFPCC method.

Epithelial-to-mesenchymal transition (EMT) facilitates the escape of pancreatic cancer cells from the primary tumor site, which is a key early event in metastasis. In the present study, we examined if intermittent hypoxia facilitates the invasiveness of human pancreatic cancer cell lines (Panc-1 and BxPC-3) by Transwell assay. We used western blotting and flow cytometry analysis to quantify stem-like cells in the migratory cells during intermittent hypoxia in the human pancreatic cancer cells. Under normoxia or intermittent hypoxia, the expression of autophagy-related proteins (LC3-II and Beclin), hypoxia-inducible factor-1α (HIF-1α) and EMT-related markers (E-cadherin, Vimentin and N-cadherin) was examined by western blotting. siRNA and the autophagic inhibitor were used to access the role of HIF-1α and autophagy in promoting metastasis and EMT. Under intermittent hypoxia, pancreatic cancer cells demonstrated enhanced invasive ability and enriched stem-like cells. The migratory cells displayed stem-like cell characteristics and elevated the expression of LC3-II and Beclin-1, HIF-1α, E-cadherin, Vimentin and N-cadherin under intermittent hypoxia conditions. Moreover, enhanced autophagy was induced by the elevated level of HIF-1α. The metastatic ability and EMT of pancreatic cancer stem cells was associated with HIF-1α and autophagy. This novel finding may indicate the specific role of HIF-1α and autophagy in promoting the metastatic ability of pancreatic cancer stem cells. Additionally, it emphasizes the importance of developing therapeutic strategies targeting cancer stem cells and autophagy to reduce metastasis.

The initiation and progression of various solid tumors, including pancreatic carcinoma, are driven by a population of cells with stem cell properties, namely cancer stem cells (CSCs). Like their normal counterparts, CSCs are also believed to rely on their own microenvironment termed niches to sustain the population. Hypoxia-inducible factor-1α (HIF-1α) is a major actor in the cell survival response to hypoxia. Recently, several researchers proposed that non-stem cancer cells can convert to stem-like cells to maintain equilibrium. The present study focuses on whether non-stem pancreatic cancer cells can convert to stem-like cells and the role of HIF-1α and autophagy in modulating this conversation. The non-stem pancreatic cancer cells and pancreatic cancer stem-like cells were separated by magnetic sorting column. Intermittent hypoxia enhanced stem-like properties of non-stem pancreatic cancer cells and stimulated the levels of HIF-1α, LC3-II and Beclin. Enhanced autophagy was associated with the elevated level of HIF-1α. The conversation of non-stem pancreatic cancer cells into pancreatic cancer stem-like cells was induced by HIF-1α and autophagy. This novel finding may indicate the specific role of HIF-1α and autophagy in promoting the dynamic equilibrium between CSCs and non-CSCs. Also, it emphasizes the importance of developing therapeutic strategies targeting cancer stem cells as well as the microenvironmental influence on the tumor.

Abstract Macrophages can be reprogramming, such as the classical activated macrophage, M1 or alternative activated macrophages, M2 phenotype following the milieu danger signals, especially inflammatory factors. Macrophage reprogramming is now considered as a key determinant of disease development and/or regression. Experimental autoimmune myocarditis (EAM) is characterized by monocytes/macrophage infiltration, Th17 cells activation and inflammatory factors producing such as high mobility group box 1 (HMGB1). Whether infiltrated macrophages could be reprogramming in EAM? HMGB1 was associated with macrophage reprogramming? Our results clearly demonstrated that infiltrated macrophage was reprogrammed towards a proinflammatory M1-like phenotype and cardiac protection by monocytes/macrophages depletion or HMGB1 blockade in EAM; in vitro , HMGB1 facilitated macrophage reprogramming towards M1-like phenotype dependent on TLR4-PI3Kγ-Erk1/2 pathway; furthermore, the reprogramming M1-like macrophage promoted Th17 expansion. Therefore, we speculated that HMGB1 contributed EAM development via facilitating macrophage reprogramming towards M1-like phenotype except for directly modulating Th17 cells expansion.

This article proposes an adaptive model-free predictive current control (MFPCC) method with optimal virtual vector modulation (OVVM) for surface-mounted permanent magnet synchronous motors. First, an ultralocal motor model based on the rotating coordinate system is established by using an adaptive extended state observer (AESO). Second, an adaptive law is devised to achieve a more balanced mitigation of disturbance and noise interference and a better transient and steady-state performance. Simultaneously, an integrated parameter estimator is designed based on the AESO, allowing for real-time adjustments of control coefficients within the ultralocal model. This dynamic calibration enhances the accuracy of model-free predictive current control and facilitates the realization of system-wide self-adaptation to handle intricate and dynamic working scenarios effectively. Third, an extended voltage vector control set based on virtual vectors is constructed to improve the accuracy of voltage selection. On this basis, apply the OVVM method to generate symmetrical pulsewidth modulation waveforms to further reduce current harmonics and improve steady-state performance. Finally, the experimental results have verified the effectiveness of the proposed MFPCC method.

Abstract Myocarditis is a common clinical cardiovascular disease, and some patients progress to dilated cardiomyopathy (DCM) with chronic heart failure. Common viral infections are the most frequent cause of myocarditis, but other pathogens and autoimmune diseases have also been implicated. Th17 cells are novel IL-17-producing effector T helper cells that play an important role in the development of autoimmune myocarditis. Furthermore, IL-17 is also important in post-myocarditis cardiac remodeling and progression to DCM. However, the mechanisms whereby IL-17 and IL-17-producing cells promote the progression of cardiac fibrosis remain unclear. We therefore investigated whether IL-17 directly induced cardiac fibrosis in experimental autoimmune myocarditis (EAM) and explored the possible molecular mechanisms. The EAM model was induced and serum IL-17 level was detected by ELISA; western blot, immunofluorescence and sirius red staining were used to analyze the collagen expression. PCR was used to assay the IL-17RA and IL-17RC. The results indicated that IL-17 induced cardiac fibrosis both in vitro and in vivo. The protein kinase C (PKC)β/Erk1/2/NF-κB (Nuclear Factor κappa B) pathway was involved in the development of myocardial fibrosis and IL-17 contributed to cardiac fibrosis following EAM via this pathway. These results provide the first direct evidence for the involvement of the PKCβ/Erk1/2/NF-κB signaling pathway in IL-17-induced myocardial fibrosis.

T helper 17(Th17) cell is a new subset of CD4(+) T cells that produce a proinflammatory cytokine interleukin-17 (IL-17). Th17 cells have recently been shown to play a critical role in many autoimmune diseases that had previously been thought to be Th1 dominant. Although Hashimoto's thyroiditis (HT) was thought to be a Th1-type disease, the contributions of Th17 cells to the pathogenesis remain unclear. In this study, we investigated the expression levels of Th1/Th17 cell-associated factors in peripheral blood mononuclear cells (PBMC) and plasma from patients with HT by quantitative real-time polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). Our results showed that the expression levels of Th1 cells-related T-bet and interferon-gamma (IFN-gamma) mRNA in PBMC from HT significantly decreased. However, the mRNA of Th17 coherent retinoic acid-related orphan nuclear receptor gamma t (RORgammat) and IL-17 in patients with HT increased. In addition, a negative correlation between T-bet and RORgammat mRNA expression was found in patients with HT, and the similar phenomena also appeared on the levels of mRNA and plasma concentration between IFN-gamma and IL-17. It suggested that Th17 cells rather than Th1 cells predominated among patients suffering from HT, and Th17 cells might be involved in the pathogenesis of HT.

Abstract High‐mobility group box 1 (HMGB1), a non‐histone nuclear protein, has been implicated in cardiovascular diseases. Dilated cardiomyopathy (DCM), one of the leading causes of heart failure, is often caused by coxsackievirus B3‐triggered myocarditis and promoted by the post‐infectious autoimmune process. Th17 cells, a novel CD4 + T subset, may be important in the pathogenesis of autoimmune myocarditis. In the present study, we attempted to block HMGB1 function with a monoclonal antibody specific for HMGB1 B box and investigated the effects of the blockade on Th17 cells and experimental autoimmune myocarditis (EAM). After induction of EAM, HMGB1 protein levels were significantly elevated both in the heart and blood. Administration of an anti‐HMGB1 B box mAb attenuated cardiac pathological changes and reduced the number of infiltrating inflammatory cells in the heart during EAM. These protective effects of HMGB1 blockade correlated with a reduced number of Th17 cells in local tissues and lower levels of IL‐17 in the serum. Furthermore, in vitro, studies demonstrated that HMGB1 promoted Th17‐cell expansion. Therefore, we speculate that HMGB1 blockade ameliorates cardiac pathological changes in EAM by suppressing Th17 cells.

Abstract Interleukin (IL)-9 belongs to the IL-2Rγc chain family and is a multifunctional cytokine that can regulate the function of many kinds of cells. It was originally identified as a growth factor of T cells and mast cells. In previous studies, IL-9 was mainly involved in the development of allergic diseases, autoimmune diseases and parasite infections. Recently, IL-9, as a double-edged sword in the development of cancers, has attracted extensive attention. Since T-helper 9 (Th9) cell-derived IL-9 was verified to play a powerful antitumor role in solid tumors, an increasing number of researchers have started to pay attention to the role of IL-9-skewed CD8 + T (Tc9) cells, mast cells and Vδ2 T cell-derived IL-9 in tumor immunity. Here, we review recent studies on IL-9 and several kinds of IL-9-producing cells in tumor immunity to provide useful insight into tumorigenesis and treatment. Graphical abstract

Rheumatoid arthritis(RA) is a common autoimmune disease associated with Th17 cells, but what about the effect of high-mobility group box chromosomal protein 1 (HMGB1) and the relationship between Th17-associated factors and HMGB1 in RA remains unknown. In the present study, we investigated the mRNA levels of HMGB1, RORγt, and IL-17 in peripheral blood mononuclear cells (PBMCs) from patients with rheumatoid arthritis by quantitative real-time PCR (RT-qPCR), and the concentrations of HMGB1, IL-17, and IL-23 in plasma were detected by ELISA. And then, the effect of HMGB1 on Th17 cells differentiation was analyzed in vitro. Our clinical studies showed that the mRNAs of HMGB1, RORγt, and IL-17 in patients were higher than that in health control (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn>0.05</mml:mn></mml:mrow></mml:math>), especially in active RA patients (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn>0.05</mml:mn></mml:mrow></mml:math>). The plasma HMGB1, IL-17, and IL-23 in RA patients were also higher than that in health control (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn>0.05</mml:mn></mml:mrow></mml:math>); there was a positive correlation between the expression levels of HMGB1 and the amount of CRP, ERS, and RF in plasma. In vitro , the IL-17-produced CD4 + T cells were increased with 100 ng/mL rHMGB1 for 12h, which indicated that the increased HMGB1 might contribute to Th17 cells activation in RA patients.

Myeloid derived suppressor cells (MDSCs) expand in cancer bearing hosts and contribute to tumor immune evasion. M2 macrophages constitute a major cellular component of cancer-related inflammation. However, the correlation between circulating MDSCs and infiltrating M2 macrophages in tumor tissues from patients with esophageal cancer (ECA), and its potential relationship with the polarization of Th2 cells remain unclear. In the present study, we showed the level of MDSCs in PBMC and Arg1 in plasma were significantly elevated in ECA patients, and the increased ratio of MDSC in PBMC was closely related to the expression of CD163 in cancer tissues. In addition, the ECA patients exhibited remarkable increases in the mRNA levels of IL-4 and GATA3, as well as the protein levels of IL-13 and IL-6, but IFN-γ and IL-12 in peripheral blood were decreased. Our data indicate that the increased Th2 cytokines are associated with MDSCs and M2 macrophages polarization, and foster the infiltration of CD163+M2 macrophages in cancer tissues, which promote the formation of immunosuppressive microenvironment in ECA patients.

Background Tumor metastasis is driven by malignant cells and stromal cell components of the tumor microenvironment. Cancer stem cells (CSCs) are thought to be responsible for metastasis by altering the tumor microenvironment. Epithelial-mesenchymal transition (EMT) processes contribute to specific stages of the metastatic cascade, promoted by cytokines and chemokines secreted by stromal cell components in the tumor microenvironment. C-C chemokine receptor 7 (CCR7) interacts with its ligand, chemokine ligand 21(CCL21), to mediate metastasis in some cancer cells lines. This study investigated the role of CCL21/CCR7 in promoting EMT and metastasis of cluster of differentiation 133+ (CD133+) pancreatic cancer stem-like cells. Methods Panc-1, AsPC-1, and MIA PaCa-2 pancreatic cancer cells were selected because of their aggressive invasive potentials. CCR7 expression levels were examined in total, CD133+ and CD133− cell fractions by Immunofluorescence analysis and real time-quantitative polymerase chain reaction (RT-qPCR). The role of CCL21/CCR7 in mediating metastasis and survival of CD133+ pancreatic cancer stem-like cells was detected by Transwell assays and flow cytometry, respectively. EMT and lymph node metastasis related markers (E-cadherin, N- cadherin, LYVE-1) were analyzed by western blot. CCR7 expression levels were analyzed by immunohistochemical staining and RT-qPCR in resected tumor tissues, metastatic lymph nodes, normal lymph nodes and adjacent normal tissues from patients with pancreatic carcinoma. Results CCR7 expression was significantly increased in CD133+ pancreatic cancer stem-like cells, resected pancreatic cancer tissues, and metastatic lymph nodes, compared with CD133− cancer cells, adjacent normal tissues and normal lymph nodes, respectively. CCL21/CCR7 promoted metastasis and survival of CD133+ pancreatic cancer stem-like cells and regulated CD133+ pancreatic cancer stem-like cells metastasis by modulating EMT and Erk/NF-κB pathway. Conclusions These results indicate a specific role for CCL21/CCR7 in promoting EMT and metastasis in CD133+ pancreatic cancer stem-like cells. Furthermore the data also indicated the potential importance of developing therapeutic strategies targeting cancer stem-like cells and CCL21/CCR7 for reducing metastasis.

Stroke is a major cause of mortality and disability worldwide. Stroke is a frequent and severe neurovascular disorder. The main cause of stroke is atherosclerosis, and the most common risk factor for atherosclerosis is hypertension. Therefore, prevention and treatment of stroke are crucial issues in humans. High mobility group box 1 (HMGB1) is non-histone nuclear protein that is currently one of the crucial proinflammatory alarmins in ischemic stroke (IS). It is instantly released from necrotic cells in the ischemic core and activates an early inflammatory response. HMGB1 may signal via its putative receptors, such as receptor for advanced glycation end products (RAGE), toll-like receptors (TLRs) as well as matrix metalloproteinase (MMP) enzymes during IS. These receptors are expressed in brain cells. Additionally, brain-released HMGB1 can be redox modified in the circulation and activate peripheral immune cells. The role of HMGB1 may be more complex. HMGB1 possesses beneficial actions, such as endothelial activation, enhancement of neurite outgrowth, and neuronal survival. HMGB1 may also provide a novel link for brain-immune communication leading to post-stroke immunomodulation. Therefore, HMGB1 is new promising therapeutic intervention aimed at promoting neurovascular repair and remodeling after stroke. In this review, we look at the mechanisms of secretion of HMGB1, the role of receptors, MMP enzymes, hypoglycemia, atherosclerosis, edema, angiogenesis as well as neuroimmunological reactions and post-ischemic brain recovery in IS. We also outline therapeutic roles of HMGB1 in IS.

Post-translational modifications (PTMs) of High mobility group box 1 (HMGB1) have not been investigated as extensively as those of other HMG proteins but accumulating evidence has shown the remarkable biological significances induced by the post-translational: acetylation, methylation and phosphorylation, oxidation, glycosylation and ADP-ribosylation of the HMGB1 to modulate its interactions with DNA and other proteins. Although HMGB1 is localized in the nucleus in almost all cells at baseline, it can be rapidly mobilized to other sites within the cell, including the cytoplasm and mitochondria, as well as into the extracellular; hence there is an increasing interest by researches into the complex relationship between the PTMs of HMGB1 protein and its diverse biological activities. The PTMs of HMGB1 could also have effects on gene expression following changes in its DNA-binding properties and in extracellular environment displays immunological activity and could serve as a potential target for new therapy. Our reviewed identifies covalent modifications of HMGB1, and highlighted how these PTMs affect the functions of HMGB1 protein in a variety of cellular and extra cellular processes as well as diseases and therapy.

High mobility group box 1 (HMGB1) is constitutively expressed by many cells. In cells, HMGB1 is a transcription factor or transcription enhancer that is involved in nucleosome sliding, DNA repair, V(D)J recombination, telomere homeostasis, autophagy and viral sensing. HMGB1 can also be secreted or released by stressed cells and serves as an alarmin, cytokine or growth factor to activate the immune response. This protein facilitates CD4+ T cell differentiation and tissue repair through binding with its receptors, including toll-like receptors (TLRs) and the receptor for advanced glycation end-products (RAGE). Recent works have established that HMGB1 plays many vital functions in cardiac inflammatory injury, cardiac regeneration and remodelling. The present review addresses the novel role of HMGB1 in secretion and cardiomyocyte senescence and in the dual faced roles of HMGB1 in cardiac inflammatory injury, inflammatory resolution and cardiac regeneration and remodelling following cardiac injury.

Cellular crosstalk is an important mechanism in the pathogenesis of inflammatory disorders and cancers. One significant means by which cells communicate with each other is through the release of exosomes. Exosomes are extracellular vesicles formed by the outward budding of plasma membranes, which are then released from cells into the extracellular space. Many studies have suggested that microvesicles released by colon cancer cells initiate crosstalk and modulate the fibroblast activities and macrophage phenotypes. Interestingly, crosstalk among colon cancer cells, macrophages and cancer-associated fibroblasts maximizes the mechanical composition of the stromal extracellular matrix (ECM). Exosomes contribute to cancer cell migration and invasion, which are critical for colon cancer progression to metastasis. The majority of the studies on colorectal cancers (CRCs) have focused on developing exosomal biomarkers for the early detection and prediction of CRC prognosis. This study highlights the crosstalk among colon cancer-derived exosomes, macrophage phenotypes and fibroblasts during colon cancer metastasis.

Phototherapy can trigger immunogenic cell death of tumors in situ, whereas it is virtually impossible to eradicate the tumor due to the intrinsic resistance and inefficient anti-tumor immunity. To overcome these limitations, novel bimetallic infinite coordination nanopolymers (TA-Fe/Mn-OVA@MB NPs) were synthesized using model antigen ovalbumin (OVA) as a template to assemble tannic acid (TA) and bi-metal, supplemented with methylene blue (MB) surface absorption. The formulated TA-Fe/Mn-OVA@MB NPs possess excellent photothermal and photodynamic therapy (PTT/PDT) performance, which is adequate to destroy tumor cells by physical and chemical attack. Especially, these TA-Fe/Mn-OVA@MB NPs are capability of promoting the dendritic cells (DCs) maturation and antigen presentation via manganese-mediated cGAS-STING pathway activation, finally activating cytotoxicity T lymphocyte and promoting memory T lymphocyte differentiation in the peripheral lymphoid organs. In conclusion, this research offers a versatile metal-polyphenol nanoplatform to integrate functional metals and therapeutic molecule for topical phototherapy and robust anti-tumor immune activation.

This article proposes a predictive cascaded speed and current control to improve the dynamic performance and robustness of permanent magnet synchronous motor drives against machine parameter variations. First, a novel model predictive speed controller is developed to achieve high performance and a wide speed range for the speed outer loop, which can minimize the system's inertial link overshoot and settling time. Second, the current inner loop utilizes an incremental current model to eliminate the complexities and variations caused by flux linkage parameters in complex working scenarios. Third, the enhanced model reference adaptive system is proposed for parameter identification, integrating the new fuzzy logic controller within the framework. Additionally, comprehensive compensation of the inverter distortion voltage is implemented to ensure improved dynamic and steady-state effectiveness, resulting in enhanced performance and higher control accuracy. Finally, experimental validation on a dSPACE system with a sampling frequency of 20 kHz demonstrates the stability and effectiveness of the proposed method.

DVL is one of the small polypeptides which plays an important role in regulating plant growth and development, tissue differentiation, and organ formation in the process of coping with stress conditions. So far, there has been no comprehensive analysis of the expression profile and function of the cotton DVL gene. According to previous studies, a candidate gene related to the development of fuzz was screened, belonging to the DVL family, and was related to the development of trichomes in Arabidopsis thaliana. However, the comprehensive identification and systematic analysis of DVL in cotton have not been conducted. In this study, we employed bioinformatics approaches to conduct a novel analysis of the structural characteristics, phylogenetic tree, gene structure, expression pattern, evolutionary relationship, and selective pressure of the DVL gene family members in four cotton species. A total of 117 DVL genes were identified, including 39 members in G. hirsutum. Based on the phylogenetic analysis, the DVL protein sequences were categorized into five distinct subfamilies. Additionally, we successfully mapped these genes onto chromosomes and visually represented their gene structure information. Furthermore, we predicted the presence of cis-acting elements in DVL genes in G. hirsutum and characterized the repeat types of DVL genes in the four cotton species. Moreover, we computed the Ka/Ks ratio of homologous genes across the four cotton species and elucidated the selective pressure acting on these homologous genes. In addition, we described the expression patterns of the DVL gene family using RNA-seq data, verified the correlation between GhMDVL3 and fuzz development through VIGS technology, and found that some DVL genes may be involved in resistance to biotic and abiotic stress conditions through qRT-PCR technology. Furthermore, a potential interaction network was constructed by WGCNA, and our findings demonstrated the potential of GhM_A05G1032 to interact with numerous genes, thereby playing a crucial role in regulating fuzz development. This research significantly contributed to the comprehension of DVL genes in upland cotton, thereby establishing a solid basis for future investigations into the functional aspects of DVL genes in cotton.

The role of integrons in the spread of antibiotic resistance has been well established. The aim of this study was to investigate the resistance profiles of Pseudomonas aeruginosa isolated from patients in Zhenjiang to 13 antibiotics, and to identify the structure and dissemination of class 1 integrons.The Kirby-Bauer disk diffusion assay was used to determine the rate of P. aeruginosa resistance. Class 1 integrons from multidrug-resistant isolates were amplified by PCR, and their PCR products were sequenced. We also analyzed the integron structures containing the same gene cassettes by restriction fragment length polymorphism (RFLP). Isolates were genotyped by pulsed-field gel electrophoresis (PFGE).The resistance rates were between 29.6% and 90.1%. The prevalence of class 1 integrons was 38.0%. These integrons included five gene cassettes (aadB, aac6-II, blaPSE-1, dfrA17, and aadA5). The dfrA17 and aadA5 gene cassettes were found most often.Class 1 integrons were found to be widespread in P. aeruginosa isolated from clinical samples in the Zhenjiang area of China. The antibiotic resistance rates in class 1 integron-positive strains of P. aeruginosa were noticeably higher than those in class 1 integron-negative strains. PFGE showed that particular clones were circulating among patients.

High-mobility group box 1 (HMGB1) is a non-histone nuclear protein that is released extracellulary and has been implicated in autoimmune disease. Toll-like receptor 2 (TLR2) signalling is thought to be essential for the inflammatory response and for immune disorders. In recent studies, enhanced HMGB1 and TLR2 expressions have been found in rheumatoid arthritis (RA), respectively. The aim of this study is to explore whether HMGB1 stimulation can up-regulate the expression of TLR2 on CD14(+) monocytes from patients with RA and to clarify the subsequent events involving Th17 cells and Th17 cell-associated cytokine changes. Our results showed that the frequency of CD14(+) cells in peripheral blood mononuclear cell (PBMC) was obviously increased, and enhanced expression of TLR2 on CD14(+) monocytes was also found in patients with RA, compared with healthy controls with statistical significance (P < 0.001). In addition, the levels of IL-17, IL-23 and IL-6 in supernatants from cultured monocytes from patients and in patient's plasma were increased, and NF-κB, the downstream target of TLR2, also showed a marked elevation after monocytes were stimulated by HMGB1. This implies that the enhanced TLR2 pathway and Th17 cell polarization may be due to HMGB1 stimulation in rheumatoid arthritis.

PPARα belongs to the peroxisome-proliferator-activated receptors (PPARs) family, which plays a critical role in inhibiting cell proliferation and tumorigenesis, while the molecular mechanism is still unclear. Here we report that PPARα serves as an E3 ubiquitin ligase to govern Bcl2 protein stability. PPARα physically bound to Bcl2 protein. In this process, PPARα/C102 was critical for PPARα binding to BH3 domain of Bcl2, subsequently, PPARα transferred K48-linked polyubiquitin to lysine-22 site of Bcl2 resulting in its ubiquitination and proteasome-dependent degradation. Importantly, overexpression of PPARα enhanced cancer cell chemotherapy sensitivity. In contrast, silenced PPARα decreased this event. These findings revealed a novel mechanism of PPARα governed endogenous Bcl2 protein stability leading to reduced cancer cell chemoresistance, which provides a potential drug target for cancer treatment.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This article has been retracted at the request of There are major errors and omissions in the description of the experiments that unfortunately were not spotted during peer review and that make it impossible to repeat the experiments or to draw meaningful conclusions based on the information given. Authors and Editors agree that a Retraction rather than a Corrigendum is the appropriate measure.

Abstract Inflammation is a critical component involved in tumor progression. Interleukin-17 (IL-17) belongs to a relatively new family of cytokines that has been associated with the progression of cancers. However, the role of IL-17B/IL-17RB (IL-17 receptor B) signaling to stemness of gastric cancer remains unknown. Here, we confirmed that the expression of IL-17RB in gastric cancer tissues was significantly increased, that overexpression was associated with poor prognosis of gastric cancer patients, and that overexpression was positively correlated with some stemness markers. Interestingly, the expression of IL-17B was upregulated in patient serum rather than gastric tumor tissues. Furthermore, exogenous rIL-17B significantly promoted the stemness of gastric cancer cells depending on IL-17RB and induced the expression of IL-17RB. Simultaneously, the expression of phosphorylated AKT, GSK-3β, and β-catenin as well as the nuclear translocation of β-catenin were significantly increased in the MGC-803 cell in a dose-dependent manner, when treated with rIL-17B. The AKT inhibitor, LY294002, and the knockdown of AKT expression reversed the rIL-17B-induced upregulation of β-catenin and some stemness markers. Together, our results indicate that the IL-17B/IL-17RB signal can promote the growth and migration of tumor cells, and upregulate cell stemness through activating the AKT/β-catenin pathway in gastric cancer, suggesting that IL-17RB may be a novel target in human gastric cancer therapy.

Gastric cancer tissue-derived mesenchymal stem cells (GC-MSCs) are important resident stromal cells in the tumor microenvironment (TME) and have been shown to play a key role in gastric cancer progression. Whether GC-MSCs exert a tumor-promoting function by affecting anti-tumor immunity is still unclear. In this study, we used GC-MSC conditioned medium (GC-MSC-CM) to pretreat peripheral blood mononuclear cells (PBMCs) from healthy donors. We found that GC-MSC-CM pretreatment markedly reversed the inhibitory effect of PBMCs on gastric cancer growth in vivo, but did not affect functions of PBMCs on gastric cancer cell proliferation, cell cycle and apoptosis in vitro. PBMCs pretreated with GC-MSC-CM significantly promoted gastric cancer migration and epithelial-mesenchymal transition in vitro and liver metastases in vivo. Flow cytometry analysis showed that GC-MSC-CM pretreatment increased the proportion of Treg cells and reduced that of Th17 cells in PBMCs. CFSE labeling and naïve CD4+ T cells differentiation analysis revealed that GC-MSC-CM disrupted the Treg/Th17 balance in PBMCs by suppressing Th17 cell proliferation and inducing differentiation of Treg cells. Overall, our collective results indicate that GC-MSCs impair the anti-tumor immune response of PBMCs through disruption of Treg/Th17 balance, thus providing new evidence that gastric cancer tissue-derived MSCs contribute to the immunosuppressive TME.

Abstract Background Acinetobacter baumannii is a multidrug-resistant (MDR) hazardous bacterium with very high antimicrobial resistance profiles. Outer membrane vesicles (OMVs) help directly and/or indirectly towards antibiotic resistance in these organisms. The present study aims to look on the proteomic profile of OMV as well as on the bacterial transcriptome upon exposure and induction with eravacycline, a new synthetic fluorocycline. RNA sequencing analysis of whole-cell and LC-MS/MS proteomic profiling of OMV proteome abundance were done to identify the differential expression among the eravacycline-induced A. baumannii ATCC 19606 and A. baumannii clinical strain JU0126. Results The differentially expressed genes from the RNA sequencing were analysed using R package and bioinformatics software and tools. Genes encoding drug efflux and membrane transport were upregulated among the DEGs from both ATCC 19606 and JU0126 strains. As evident with the induction of eravacycline resistance, ribosomal proteins were upregulated in both the strains in the transcriptome profiles and also resistance pumps, such as MFS, RND, MATE and ABC transporters. High expression of stress and survival proteins were predominant in the OMVs proteome with ribosomal proteins, chaperons, OMPs OmpA, Omp38 upregulated in ATCC 19606 strain and ribosomal proteins, toluene tolerance protein, siderophore receptor and peptidases in the JU0126 strain. The induction of resistance to eravacycline was supported by the presence of upregulation of ribosomal proteins, resistance-conferring factors and stress proteins in both the strains of A. baumannii ATCC 19606 and JU0126, with the whole-cell gene transcriptome towards both resistance and stress genes while the OMVs proteome enriched more with survival proteins. Conclusion The induction of resistance to eravacycline in the strains were evident with the increased expression of ribosomal and transcription related genes/proteins. Apart from this resistance-conferring efflux pumps, outer membrane proteins and stress-related proteins were also an essential part of the upregulated DEGs. However, the expression profiles of OMVs proteome in the study was independent with respect to the whole-cell RNA expression profiles with low to no correlation. This indicates the possible role of OMVs to be more of back-up additional protection to the existing bacterial cell defence during the antibacterial stress.

The myocardium responds to aetiologically different pathological injuries through a common multistep process involving highly co-ordinated interactions between cardiac and immune cells. Cardiac fibroblast cells which constitute the prevalent cell type in the heart to have their functional effects that express contractile proteins and exhibit increased migratory, proliferative and secretory properties. During the pathogenesis of myocarditis, cardiac fibroblast, dendritic cells, macrophages, CD4(+) T cells and other immune cells are known to play variable roles. It is becoming increasingly clear that cardiac fibroblasts are not passive players in immune responses, and several evidences show this through the release of soluble signals and/or direct interactions with these immune cells. Typically, fibroblasts are involved in synthesizing factors such as cytokines, chemokines, prostanoids, matrix components and matrix-degrading enzymes to influence dendritic cells, CD4(+) T cells and macrophage functions and vice versa in the pathogenesis of myocarditis. Again, evidence proves a crosstalk between cardiac fibroblasts and immune cells recruited into the myocardium during myocarditis in the microenvironments. This piece reviews the properties and roles of cardiac fibroblast cells, dendritic cells, macrophages and CD4(+) T cells in the pathogenesis of myocarditis, and how these cells interplay on each other in the microenvironment.

Cervical cancer is one of the most common cancers in women worldwide, often associated with the infection of human papillomavirus (HPV). Toll-like receptor 8 (TLR8), a pattern recognition receptor, is involved in viral nucleic acid sensing. Recently TLR8 has been shown to be expressed in cancer cells, and it has been suggested that it may help cancer cell growth and tumor development. The objective of this study is to investigate the expression of TLR8 expression and its relationship with Bcl-2 and VEGF in cervical cancer cells.The mRNA expression levels of Bcl-2, VEGF and TLR-7,-8,-9 in newly diagnosed cervical cancer patients were detected by quantitative real-time PCR (qRT- PCR). Epifluorescence microscope was used to determine the presence of TLR8 protein in Hela cells. The cell cycle and apoptosis were analyzed by flow cytometer, and the cell proliferation was measured by MTT assay. Our data showed the increased mRNA levels of TLR8 in human cervical cancer samples as well as in HeLa cells, a cell line derived from a human cervical cancer. In addition, there was a positive correlation between the expression levels of TLR8 and Bcl-2 and VEGF in cervical cancer patients. When Hela cells were treated with TLR8 agonist CL075, the percentage of cells in G2/M +S was remarkably increased, accompanied by increased COX-2, BCL-2 and VEGF mRNA levels.The mRNA expression level of TLR8 in the patients with cervical cancer and Hela cells were up-regulated, it consistent with the increased expression of VEGF and Bcl-2. The results suggest that TLR8 may be an interesting therapeutic target in cervical cancer.

Abstract Upregulated high‐mobility group box 1 ( HMGB 1) has been found in many diseases. Nevertheless, the function of HMGB 1 on modulating the proliferation of lung cancer cells (Lewis cells) and inhibiting apoptosis is poorly understood, as well as the involved intracellular signalling. In the present study, we firstly found the apoptosis of Lewis was increased following Hanks’ balanced salt solution ( HBSS )‐induced starvation, while it was rescued after exogenous HMGB 1 protein was added; furthermore, the receptor for advanced glycation end products ( RAGE ) and T oll‐like receptor ( TLR 4) could coordinately improve the proliferation of tumour cells in vitro, and HMGB 1 could enhance the phosphorylation of PI 3 K / A kt and E rk1/2, inhibit the expression of pro‐apoptosis protein B ax and promote the expression of anti‐apoptosis protein B cl‐2. These findings clearly demonstrated that HMGB 1– RAGE / TLR 4‐ PI 3 K ‐ A kt/ E rk1/2 pathway contributed to the proliferation of Lewis. Moreover, our observations provide experimental and theoretical basis for clinical biological therapy for cancers; it also may be a new target for intervention and treatment of lung cancer.

Autophagy is a self-digesting mechanism responsible for the removal of long-lived proteins and damaged organelles by lysosomes. It also allows cells to survive during nutrient depletion and/or in the absence of growth factors. High-mobility group protein 1 (HMGB1) is a highly-conserved nuclear protein that has been associated with cell autophagy; however, the mechanisms responsible for this role remain unclear. Many reports have demonstrated that autophagy represents a survival strategy for tumor cells during nutrient depletion, oxidative stress and DNA damage. In the present study, we explored the mechanisms whereby HMGB1 regulates tumor cell autophagy during nutrient depletion (the cells were cultured in Hank's balanced salt solution, HBSS). HMGB1 expression in Lewis cells increased and the protein was shuttled from the nucleus to the cytoplasm and was secreted, coincident with up-regulation of autophagy. Prevention of HMGB1 binding to the receptor for advanced glycation end products (RAGE) or knock-down of HMGB1 expression led to inhibition of autophagy and increased apoptosis. These results demonstrated a positive feedback pathway whereby starvation of Lewis cells promoted HMGB1 secretion, allowing cells to survive by regulating autophagy via a RAGE–HMGB1-extracellular signal-regulated kinase1/2-dependent pathway. These results also implicate HMGB1 as a potential risk factor for cancer growth and metastasis.

IFN-γ-producing Th17 cells have been implicated in autoimmune disorders, but their properties in humans are known only partially. The molecular mechanisms and external factors that govern IFN-γ-producing Th17-cell bias are incompletely understood. The present work was to clarify whether (i) IFN-γ-producing Th17 cells are present in the peripheral circulation of patients with coronary atherosclerosis (CA); (ii) high mobility group box (HMGB)1 in circulation is associated with IFN-γ-producing Th17-cell bias.Thirty-six patients (17 females and 19 males; 45-84 years) diagnosed as having atherosclerosis after coronary angiography for suspected or known CA were included the study cohort. Samples of peripheral blood were collected from healthy volunteers and patients, and classical tests (flow cytometry, RT-qPCR) were used to measure blood components.Our results clearly demonstrated that HMGB1 were up-regulated in different progressive CA patients: 5.38 ± 1.48 ng/ml, 6.30 ± 1.53 ng/ml and 5.86 ± 1.12 ng/ml vs1.45 ± 0.65 ng/ml for only atherosclerotic plaque (AP), atherosclerotic plaque and some plaque rupture, no thrombosis (PR), plaque rupture and accompanying thrombosis (TH) and volunteers, respectively, p < 0.05. The frequency of IFN-γ-producing Th17 cells was 2.33 ± 0.58%, 1.93 ± 0.2% and 2.21 ± 0.65% vs 0.38 ± 0.21% for AP, PR, TH and volunteers, p < 0.05, respectively. Furthermore, HMGB1 contributed to IFN-γ-producing Th17-cell bias by controlling expression of T-bet and RUNX3. We demonstrated, for the first time, that HMGB1 is a potential inducer of IFN-γ-producing Th17-cell bias, and that IFN-γ-producing Th17 cells might be one of the pathogenic factors in atherosclerosis.

Gliomas represent 60% of primary intracranial brain tumors and 80% of all malignant types, with highest morbidity and mortality worldwide. Although glioma has been extensively studied, the molecular mechanisms underlying its pathology remain poorly understood. Clarification of the molecular mechanisms involved in their development and/or treatment resistance is highly required. High mobility group box 1 protein (HMGB1) is a nuclear protein that can also act as an extracellular trigger of inflammation, proliferation and migration, through receptor for advanced glycation end products and toll like receptors in a number of cancers including gliomas. It is known that excessive release of HMGB1 in cancer leads to unlimited replicative potential, ability to develop blood vessels (angiogenesis), evasion of programmed cell death (apoptosis), self-sufficiency in growth signals, insensitivity to inhibitors of growth, inflammation, tissue invasion and metastasis. In this review we explore the mechanisms by which HMGB1 regulates apoptosis and autophagy in glioma. We also looked at how HMGB1 mediates glioma regression and promotes angiogenesis as well as possible signaling pathways with an attempt to provide potential therapeutic targets for the treatment of glioma.

Over the past decade, studies have identified subset of B cells, which play suppressive functions in additions to the conventional functions of B cells: antigen processing and presentation, activation of T cells and antibody productions. Because of their regulatory function, they were named as B regulatory cells (Bregs). Bregs restrict the severity of autoimmune disorders in animal disease models such as experimental autoimmune myocarditis (EAM), experimental autoimmune encephalitis (EAE), and collagen-induced arthritis (CIA) but can contribute to the development of infection and cancer. In humans, the roles of B regulatory cells in autoimmune diseases have not been clearly established because of the inconsistent findings from many researchers. This is believed to arise from the speculated fact that Bregs lack specific marker, which can be used to identify and characterize them in human diseases. The CD19+CD24hiCD38hiCD1dhiB cells have been associated with the regulatory function. Available evidences highlight the relevance of increasing IL-10-producing B cells in autoimmune diseases and the possibility of serving as new therapeutic targets in inflammatory disorders. This review empanels the functions of Bregs in autoimmune diseases in both human and animal models, and further evaluates the possibility of Bregs as therapeutic targets in inflammatory disorders. Consequently, this might help identify possible research gaps, which need to be clarified as researchers speculate the possibility of targeting some subsets of Bregs in the treatment of inflammatory disorders.

Abstract Enterovirus 71 (EV71) affects the health of young children globally causing severe neurologic diseases. The relationship between EV71 infection and T helper type 17 (Th17) has not been described, although this new Th subset or interleukin‐17 (IL‐17) has been reported to be associated with other viral infections. The purpose of the current study was to describe the immune profile involving Th17 cells, neutrophils, and related factors and to speculate on the possible immunopathogenesis of EV71 infections. Flow cytometry and an automatic blood cell counter were used to analyze circulating Th17 cells and count neutrophils, respectively. Expression of acid‐related orphan nuclear receptor gamma t (ROR γt) was evaluated by reverse‐transcriptional PCR, and enzyme linked immunosorbent assays (ELISAs) were used for detecting concentrations of IL‐17, IL‐23, and IFN‐γ. The results showed that the frequencies of Th17 cells (1.47 ± 0.87%) and the number of neutrophils (7.4 ± 4.1 × 10 9 /L) in peripheral blood samples from children infected with EV71 were significantly higher compared to controls. In addition, there was a statistically higher expression of ROR γt in peripheral blood mononuclear cells (PBMCs) and elevated concentrations of IL‐17 and IL‐23 in sera, but lower IFN‐γ production during EV71 infections. The findings suggest that Th17 cells are mediators during the immunologic process. J. Med. Virol. 84:763–767, 2012. © 2012 Wiley Periodicals, Inc.

In pregnancy, the immunologic system plays an important role that ensures normal pregnancy development and can as well promote the development of complications. Pregnancy success appears to rely on a discrete balance between the Th cytokines, which are involved in fetal growth and development. Preeclampsia and gestational diabetes are known complications associated with pregnancy. However, the source of the increased IL-17 cytokine in preeclampsia and other pregnancy associated diseases still remains unclear amidst numerous inconsistencies. The recent identification of innate lymphoid cells (ILC) has raised more doubts about the sources of most of the Th associated cytokines. We investigated the source of peripheral IL-17 levels in preeclamptic, gestational diabetics and chronic diabetics compared to healthy pregnancy subjects. To evaluate the source of the increased IL-17 cytokine among preeclampsia, chronic diabetic and gestational diabetic patients we investigated the proportion of Th17 cell populations in peripheral blood mononuclear cells using flow cytometry as well as analyzing levels of IFN-γ, IL-17, IL-1β and HMGB1. This study found that the Th17 cell populations in peripheral blood of preeclamptic, gestational nor chronic diabetes during pregnancy did not correlate with the increased IL-17. We report that the increased IL-17 levels observed in patients with preeclampsia, gestational diabetes and chronic diabetes are associated with innate lymphoid cells 3 (ILC3) and may pose threats to the fetus if disregulated.

High mobility group box 1 (HMGB1), an important inflammatory mediator, is actively secreted by immune cells and some non-immune cells or passively released by necrotic cells. HMGB1 has been implicated in many inflammatory diseases. Our previous published data demonstrated that HMGB1 was up-regulated in heart tissue or serum in experimental autoimmune myocarditis (EAM); HMGB1 blockade could ameliorate cardiac fibrosis at the last stage of EAM. And yet, until now, no data directly showed that HMGB1 was associated with cardiac fibrosis. Therefore, the aims of the present work were to assess whether (1) up-regulated HMGB1 could directly lead to cardiac fibrosis in EAM; (2) cardiac fibroblast/myofibroblasts could secrete HMGB1 as another source of high-level HMGB1 in EAM; and (3) HMGB1 blockade could effectively prevent cardiac fibrosis at the last stage of EAM. Our results clearly demonstrated that HMGB1 could directly lead to cardiac collagen deposition, which was associated with PKCβ/Erk1/2 signalling pathway; furthermore, cardiac fibroblast/myofibroblasts could actively secrete HMGB1 under external stress; and HMGB1 secreted by cardiac fibroblasts/myofibroblasts led to cardiac fibrosis via PKCβ activation by autocrine means; HMGB1 blockade could efficiently ameliorate cardiac fibrosis in EAM mice.

Abstract The high‐mobility group box‐1 (HMGB1), as a highly conserved ubiquitous DNA‐binding protein, has been widely studied in various diseases, including inflammation and tumor; however, fewer studies were focused on the mechanisms controlling HMGB1 release compared with the function of HMGB1. Previous studies have proven that ANG II can act as a pro‐inflammatory cytokine, both of HMGB1 and ANG II were significantly upregulated in autoimmune diseases; however, the exact role of ANG II in regulating HMGB1 release have not been shown. The present study was to define the effects of ANG II on macrophages and the possible mechanisms in controlling HMGB1 release. Our results showed that ANG II can induce M1 macrophage polarization through upregulated the expression of HMGB1 and caused acetylation of HMGB1 and release via its dissociation from SIRT1, which in a positive feedback upregulates ANG II. Subsequently, HMGB1 inhibitors can reduce the ANG II‐elicited polarize of macrophage. Meanwhile, we show that JAK/STAT pathways play an essential role in ANG II‐induced HMGB1 nuclear translocation, JAK/STAT specific inhibitors can inhibit ANG II‐induced HMGB1 expression. Taken together, our results provide a novel evidence that HMGB1 play a critical role in ANG II mediated macrophage polarization, and we suggest that ANG II mediated HMGB1 release via dissociation from SIRT1, induce hyperacetylation of HMGB1, thus for subsequent release, suggesting that the angiotensin II receptor antagonist is a potential drug target for inhibiting HMGB1 release in inflammation diseases.

Interleukin-22 (IL-22), which belongs to the IL-10 family, is an alpha helix cytokine specifically produced by many lymphocytes, such as Th1, Th17, Th22, ILCs, CD4+ and CD8+ T cells. In recent years, more and more studies have demonstrated that IL-22 has an interesting relationship with various cardiovascular diseases, including myocarditis, myocardial infarction, atherosclerosis, and other cardiovascular diseases, and IL-22 signal may play a dual role in cardiovascular diseases. Here, we summarize the recent progress on the source, function, regulation of IL-22 and the effects of IL-22 signal in cardiovascular diseases. The study of IL-22 will suggest more specific strategies to maneuver these functions for the effective treatment of cardiovascular diseases and future clinical treatment.

Asthma is a disease of the respiratory system that is commonly considered a T-helper 2 (Th2) cell-associated inflammatory disease. Group 2 innate lymphoid cells (ILC2s) promote the inflammatory responses in asthma by secreting type 2 cytokines. Interleukin (IL)-9 also serves as a promoting factor in asthma and it is well known that ILC2s have an autocrine effect of IL-9 to sustain their survival and proliferation. However, the specific role of ILC2-derived IL-9 in asthma remains unclear. HMGB1 (High-Mobility Group Box-1) is a nuclear protein, and Previous studies have shown that HMGB1 can regulate the differentiation of T-helper cells and participate in the development of asthma. But whether HMGB1 can regulate the innate lymphocytes in the pathological process of asthma is unknown. In this study we have shown increased presence of HMGB1 protein in the lung of mice with asthma, which was associated with increased secretion of IL-9 by ILC2s. This led to the activation of dendritic cells (DCs) that can accelerate the differentiation of Th2 cells and worsen the severity of asthma. Taken together, our study provides a complementary understanding of the asthma development and highlights a novel inflammatory pathway in the pathogenesis of asthma

Summary Gastric cancer is a serious public health cancer and causes nearly 1 million deaths a year worldwide. Th1 cells play critical roles in orchestrating the adaptive immune responses against gastric cancer. T‐bet , a member of the T‐box family of transcription factors, is the Th1 master regulator and up‐regulated during Th1 differentiation. Polymorphisms have also been shown to exist in T‐bet. Some reports indicated that some tumours were associated with the drift of Th1 and Th2. In the present work, we investigated the drift of Th1/Th2 by detecting the expression levels of T‐bet , IFN‐γ, IL‐4, and GATA‐3 in peripheral blood mononuclear cell of gastric cancer patients by real‐time PCR, explored the relationship between the polymorphism of T‐bet gene and drift of Th1/Th2 by gene sequence, western blot, and gene transfection. Our results indicated that a predominant Th2 phenotype was existence. T‐bet gene mutations may be associated with Th2‐dominated condition in gastric cancers.

Group 2 innate lymphoid cells (ILC2s) were demonstrated to be involved in the initiation and coordination of type 2 T helper cell (Th2) responses. Myeloid‑derived suppressor cells (MDSCs) have received a great deal of attention for their role in creating an immunosuppressive microenvironment in cancer‑bearing hosts. However, the contributions of ILC2s in the occurrence and development of lung cancer, and the association between ILC2s and Th2 or MDSCs in lung cancer remain to be elucidated. In the present study, 36 patients newly diagnosed with lung cancer based on the guidelines of the International Union Against Cancer Tumor Node Metastasis were included. The frequencies of ILC2s and MDSCs in peripheral blood mononuclear cells were determined, and the mRNA expression levels of ILC2s or Th2‑related transcription factors and cytokines, and MDSCs‑related products were assessed. The results demonstrated that the frequencies of the circulatory ILC2s and MDSCs were enhanced in lung cancer patients, as were ILC2‑related transcription factors and cytokines in peripheral blood. A positive correlation was identified between the Th2‑dominated phenotype and the expression levels of ILC2s‑associated cytokines or transcription factors. In addition, increased autophagy related 1 was closely associated with Th2‑associated transcription factors. It was demonstrated that ILC2s and MDSCs were clearly upregulated and accompanied by a predominant Th2 phenotype in patients with lung cancer; this may lead to new immunotherapy approaches for lung cancer based on the associated metabolites and cytokines.

B10 cells, a specific subset of regulatory B cells, are capable of regulating immune response and restricting inflammation and autoimmune disease progression by producing IL-10. B10 cells frequently change significantly during inflammation and autoimmunity. However, how B10 cell populations change in viral myocarditis (VMC) remains unclear. Therefore, this work was conducted to clarify the changes in B10 cells and their potential mechanisms. Our results showed that the B10 cell frequency significantly changed in the VMC model. Changes in prostaglandin E2 (PGE2) levels in VMC model hearts were consistent with B10 expansion. PGE2 induced B10 cell expansion via the MAPKs/AKT-AP1 axis or AhR signaling. Additionally, PGE2-pretreated B10 cells inhibited naïve CD4+ T cell differentiation into Th17 cells. In vivo, PGE2 treatment or adoptive B10 cell transfer significantly restricted VMC development. Our results provide sufficient evidence that PGE2-induced B10 cell expansion may become a promising therapeutic approach for VMC and acute inflammatory injury.

Acinetobacter baumannii, as a nonfermentation Gram-negative bacterium, mainly cause nosocomial infections in critically ill patients. With the widespread of multidrug-resistant Acinetobacter baumannii, the urgency of developing effective therapy options has been emphasized nowadays. Outer membrane vesicles derived from bacteria show potential vaccine effects against bacterial infection in recent study. Our present research is aimed at investigating the mechanisms involved in immune protection of mice after outer membrane vesicle immunization. As our data showed, the outer membrane vesicle from an Acinetobacter baumannii clinical strain could activate bone marrow-derived dendritic cells (BMDCs) to promote Th2 activity together with humoral immune responses to Acinetobacter baumannii-induced sepsis, which might enlighten people to have a better understanding of OMVs' role as a vaccine to prevent bacterial infections.

Abstract Resident cardiac macrophages play important roles in homeostasis, maintenance of cardiac function, and tissue repair. After cardiac injury, monocytes infiltrate the tissue, undergo phenotypic and functional changes, and are involved in inflammatory injury and functional remodelling. However, the fate of cardiac infiltrating/polarized macrophages and the relationship between these cells and resident cardiac macrophage replenishment following injury remain unclear. Our results showed that angiotensin II induces cardiac fibroblast transdifferentiation into cardiac myofibroblasts (MFBs). In cocultures with MFBs and murine macrophages, the MFBs promoted macrophage polarization to M1 phenotype, followed by selective apoptosis, which was associated with TNF/TNFR1 axis and independent of NO production. Surprisingly, after 36 h of coculture, the surviving macrophages were converted to M2 phenotype and settled in heart, which was dependent on leptin produced by MFBs or polarized macrophages via the PI3K or Akt pathway. CCR2 + CD45.2 + cells adoptively transferred into CD45.1 + mice with viral myocarditis, differentiated into CD45.2 + CCR2 + CX3CR1 + M2 cells during the resolution of inflammation and settled within the heart. Our data highlight a novel mechanism related to the renewal or replenishment of cardiac resident macrophages following cardiac injury; and suggest that transdifferentiation of cardiac fibroblasts may promote the resolution of inflammation.

Safe and potent adjuvants are required to establish effective vaccines. In the present work, we show that calcineurin subunit B promotes the secretion of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin-12p70 (IL-12 p70), IL-6 and the chemokine IL-8. It also up-regulates transcript levels of chemokines in bone marrow-derived dendritic cells. In an animal model, C57BL/6 mice were divided into four groups, immunized with ovalbumin (OVA), OVA mixed with calcineurin subunit B (CnB), CnB and PBS, respectively. The splenocytes from mice immunized with OVA in combination with CnB produced higher levels of IFN-γ and CTL when in vitro stimulated with OVA protein. Subcutaneous (s.c.) immunization of C57BL/6 mice with OVA plus CnB conferred greater protection against tumor-forming E.G7-OVA cells than did injection of OVA alone, and the survival rate of mice immunized intraperitoneally was higher than that of mice immunized s.c. Thus, CnB exerts potent adjuvant effects that polarize responses toward T(h)1 and elicit anti-tumor and anti-infection immunity. CnB could be of use as a prophylactic adjuvant as it is a human-derived safe agent and has immune-modulating effects that promote the control of cancer and infectious diseases.

Myeloid-derived suppressor cells (MDSCs) were originally described as a heterogeneous population of immature cells derived from myeloid progenitors with immune-suppressive functions in tumor-bearing hosts. In recent years, increasing number of studies have described various populations of myeloid cells with MDSC-like properties in murine models of cancer and autoimmune diseases. These studies have observed that the populations of MDSCs are increased during inflammation and autoimmune conditions. In addition, MDSCs can effectively suppress T cell responses and modulate the activity of natural killer cells and other myeloid cells. MDSCs have also been implicated in the induction of regulatory T cell production. Furthermore, these cells have the potential to suppress the autoimmune response, thereby limiting tissue injury. Myeloid regulatory cells (Mregs) are recently attracting increasing attention, since they function in proinflammatory and immune suppression in autoimmune diseases, as well as in various types of cancer. Currently, research focus is directed from MDSCs to Mregs in cancer and autoimmune diseases. The present study reviewed the suppressive roles of MDSCs in various autoimmune murine models, the immune modulation of MDSCs to T helper 17 lymphocytes, as well as the proinflammatory and immunosuppressive roles of Mregs in various types of cancer and autoimmune diseases.

The process of host infection by bacteria is complicated. Bacterial infections strongly induce the host immune system, which necessitates a robust clearance of the infection. However, bacteria have over time developed strategies that enable their evasion of attacks by the host immune system. One such strategy is the type VI secretion system (T6SS), a special needle-like secretion system that is widespread in Gram-negative bacteria and is responsible for delivering effector proteins into the external bacterial environment or directly into the host cell cytosol. Bacterial T6SS and its secreted effector proteins play an important role in the interaction between bacteria and host immune system. They also serve as antigens that are employed in the development of vaccines for clinical trials as well as future vaccine candidates. This review focuses mainly on aspects of T6SS effectors that impact the strength of the host immune system, including inflammation, autophagy, and apoptosis (silent programmed cell death). The T6SS-based vaccines are also described.

Abstract Bufalin, a major cardiotonic compound of the traditional Chinese medicine Chanshu has been used for cancer treatment for several years. However, the molecular mechanisms of Bufalin‐induced autophagy in osteosarcoma (OS) is not fully understood. In the present study, it was shown that Bufalin induced crosstalk between apoptosis and autophagy, which resulted in OS cell death. Mechanistically, Bufalin induced autophagy by increased the ratio of microtubule‐associated protein 1A/1B‐light chain 3 (LC3)‐II/LC3‐I, and inducing apoptosis via the caspase‐dependent pathway. Inhibition of autophagy promoted Bufalin‐induced cell death. In contrast, suppression of apoptosis enhanced Bufalin‐induced autophagy. In addition, it was found that Bufalin activated the Ca 2+ /calmodulin‐dependent protein kinase β/AMPK/Beclin1 pathway, which resulted in induction of autophagy. These findings provide a mechanistic understanding of the means by which Bufalin mediates autophagy and apoptosis in OS cells.

Sulfotransferases (SOTs) (EC 2.8.2.-) are sulfate regulatory proteins in a variety of organisms that have been previously shown to be involved in regulating a variety of physiological and biological processes, such as growth, development, adaptation to land, stomatal closure, drought tolerance, and response to pathogen infection. However, there is a lack of comprehensive identification and systematic analysis of SOT in cotton, especially in G. barbadense. In this study, we used bioinformatics methods to analyze the structural characteristics, phylogenetic relationships, gene structure, expression patterns, evolutionary relationships, selection pressure and stress response of SOT gene family members in G. barbadense. In this study, a total of 241 SOT genes were identified in four cotton species, among which 74 SOT gene members were found in G. barbadense. According to the phylogenetic tree, 241 SOT protein sequences were divided into five distinct subfamilies. We also mapped the physical locations of these genes on chromosomes and visualized the structural information of SOT genes in G. barbadense. We also predicted the cis-acting elements of the SOT gene in G. barbadense, and we identified the repetitive types and collinearity analysis of SOT genes in four cotton species. We calculated the Ka/Ks ratio between homologous gene pairs to elucidate the selective pressure between SOT genes. Transcriptome data were used to explore the expression patterns of SOT genes, and then qRT-PCR was used to detect the expression patterns of GBSOT4, GBSOT17 and GBSOT33 under FOV stress. WGCNA (weighted gene co-expression network analysis) showed that GB_A01G0479 (GBSOT4) belonged to the MEblue module, which may regulate the resistance mechanism of G. barbadense to FOV through plant hormones, signal transduction and glutathione metabolism. In addition, we conducted a VIGS (virus-induced gene silencing) experiment on GBSOT4, and the results showed that after FOV inoculation, the plants with a silenced target gene had more serious leaf wilting, drying and cracking than the control group, and the disease index of the plants with the silenced target gene was significantly higher than that of the control group. This suggests that GBSOT4 may be involved in protecting the production of G. barbadense from FOV infection. Subsequent metabolomics analysis showed that some flavonoid metabolites, such as Eupatorin-5-methylether (3′-hydroxy-5,6,7,4′-tetramethoxyflavone, were accumulated in cotton plants in response to FOV infection.

Intercropping legume with grass has potential to increase biomass and protein yield via biological N2-fixation (BNF) benefits, whereas the joint effects of biochar (BC) coupled with deficit irrigation on intercropping systems remain elusive. A 15N isotope-labelled experiment was implemented to investigate morpho-physiological responses of faba bean-ryegrass intercrops on low- (550 °C, LTBC) or high-temperature BC (800 °C, HTBC) amended sandy-loam soil under full (FI), deficit (DI) and partial root-zone drying irrigation (PRD). LTBC and HTBC significantly reduced intrinsic water-use efficiency (WUE) by 12 and 14 %, and instantaneous WUE by 8 and 16 %, respectively, in faba bean leaves, despite improved photosynthetic (An) and transpiration rate (Tr), and stomatal conductance (gs). Compared to FI, DI and PRD lowered faba bean An, gs and Tr, but enhanced leaf-scale and time-integrated WUE as proxied by the diminished shoots Δ13C. PRD enhanced WUE as lower gs, Tr and guard cell length than DI-plants. Despite higher carbon ([C]) and N concentration ([N]) in faba bean shoots amended by BC, the aboveground C- and N-pool of faba bean were reduced, while these pools increased for ryegrass. The N-use efficiency (NUE) in faba bean shoots was reduced by 9 and 14 % for LTBC and HTBC, respectively, but not for ryegrass. Interestingly, ryegrass shoots had 52 % higher NUE than faba bean shoots. The N derived from atmosphere (% Ndfa) was increased by 2 and 9 % under LTBC and HTBC, respectively, while it decreased slightly by reduced irrigation. Quantity of BNF in faba bean aboveground biomass decreased with HTBC coupled with reduced irrigation, mainly towards decreased biomass and soil N uptake by faba bean. Therefore, HTBC might not be a feasible option to improve WUE and BNF in faba bean-ryegrass intercropping, but PRD is permissible as the clear trade-off between BC and PRD.

[This corrects the article DOI: 10.1371/journal.pone.0059604.].

The outer membrane protein RagB is one of the major virulence factors of the periodontal pathogen Porphyromonas gingivalis (P. gingivalis). In order to induce protective immune response against P. gingivalis infection, an mGITRL gene-linked ragB DNA vaccine (pIRES-ragB-mGITRL ) was constructed. Six-week-old female BALB/c mice were immunized with pIRES-ragB-mGITRL through intramuscular injection and then challenged by subcutaneous injection in the abdomen with P. gingivalis. RagB-specific antibody-forming cells were evaluated by an Enzyme-linked immunosorbent spot, and specific antibody was determined by enzyme-linked immunosorbent assay. In addition, the frequencies of Tfh and IFN-γ+ T cells in spleen were measured using flow cytometer, and the levels of IL-21 and IFN-γ mRNA or proteins were detected by real time RT-PCR or ELISA. The data showed that the mGITRL-linked ragB DNA vaccine induced higher levels of RagB-specific IgG in serum and RagB-specific antibody-forming cells in spleen. The frequencies of Tfh and IFN-γ+ T cells were obviously expanded in mice immunized by pIRES-ragB-mGITRL compared with other groups (pIRES or pIRES-ragB ). The levels of Tfh and IFN-γ+ T cells associated cytokines were also significantly increased in pIRES-ragB-mGITRL group. Therefore, the mice immunized with ragB plus mGITRL showed the stronger resistant to P. gingivalis infection and a significant reduction of the lesion size caused by P. gingivalis infection comparing with other groups. Taken together, our findings demonstrated that intramuscular injection of DNA vaccine ragB together with mGITRL induced protective immune response dramatically by increasing Tfh and IFN-γ+ T cells and antibody production to P. gingivalis.

High mobility group box 1 (HMGB1), a non-histone nuclear protein, was associated with a variety of biological important processes, such as transcription, differentiation, extracellular signaling. As a cytokine or inflammatory mediator, more and more data showed that HMGB1 was involved in inflammatory diseases, cancers or autoimmune disease. However, few data focused on nucleic or cytoplasmic function of HMGB1. Therefore, the present study focused on cancer cells biological characteristics following HMGB1 silence. HMGB1 siRNAs were designed and chemically synthesized, and then transfected into the breast cancer cell line MCF-7 with lipofectamine 2000. The transcription and translation level of HMGB1 expression, proliferation, apoptosis, migration of MCF-7 were determined. The results demonstrated that HMGB1 silence inhibit invasion and migration and promote apoptosis of human breast cells; which indicated that HMGB1 silence might be a potential therapy targets.

Myeloid-derived suppressor cell (MDSC) is an important heterogeneous cell population that regulates innate and adaptive immune. The immunosuppressive potentiality is widely known. However, more and more data indicated that the heterogeneous cell population had immunostimulatory activities rather than immunosuppression in some microenvironments. Chronic inflammatory conditions contribute to accumulation of MDSCs. These pathologically activated cells played critical roles in cancer and inflammatory diseases. In this review, we mainly discuss the immunoregulation potentiality of MDSCs in cancer and inflammatory diseases.

Cardiac-resident macrophages (CRMs) play critical roles in maintaining cardiac homoeostasis and removing senescent and dying cells. Recent preclinical data have re-energised the area of cardioimmunology and provided improved understanding of the modulation of compositional and functional phenotypes of CRMs. These data can aid in achieving improved cardiac regeneration, repair and functional remodelling following cardiac injury. In this review, we discuss the composition and renewal of various subsets of CRMs. Specific attention has been given to delineate the roles of various CRM subsets with respect to (1) facilitation of cardiac development and maintenance of physiological function such as electrical conduction and rhythm; (2) promotion of cardiac regeneration, inflammation resolution and functional remodelling following a cardiac injury; and (3) therapeutic potential. We have also highlighted the relationship between CRM replenishment and cardiomyocyte senescence as well as cardiovascular diseases development. Finally, we have addressed future perspectives and directions in basic research and potentially clinical applications of CRMs.

To determine the rate of furazolidone resistance of Helicobacter pylori (H. pylori) isolated from gastric biopsy specimens and to explore the relationship between genetic mutations in porD and oorD genes of H. pylori and its resistance to the antibiotic.Gastric biopsy was performed in 83 adult patients aged 31-77 years with gastric complaints. H. pylori was isolated from biopsy specimens of 46 patients. E-test and 2-fold agar dilution method were used to determine the rate of H. pylori resistance to furazolidone. The genes porD and oorD from susceptible and resistant isolates were amplified by polymerase chain reaction (PCR), and their PCR products were sequenced.Resistance to furazolidone was found in 8.7% of H. pylori isolates and 6 mutations were detected in porD and oorD genes of the resistant isolates. Three mutations--G353A, A356G, and C357T--occurred in porD and the other mutations--A041G, A122G, C349A(G)--occurred in oorD genes.Changes in 6 amino acids may be associated with the resistance of H. pylori to furazolidone.

Group 2 innate lymphoid cells (ILC2s) are considered to be the most significant mediators during the orchestration of immune responses in asthma. Myeloid-derived suppressor cells (MDSCs) has received a great deal of attention for their immunosuppressive activity, and our early studies indicate that the increased Th2 cytokines are associated with MDSCs. In this study, we sought to determine whether MDSCs are also participation in immune imbalance and its relationship with ILC2s in asthma. The data showed that the circulatory ILC2s or MDSCs and their characteristic cytokines or transcription factors were significantly enhanced in asthmatic patients, as well as in chronic obstructive pulmonary disease (COPD) or respiratory viral infections (RVI). Meanwhile, a Th2-dominated phenotype was found in patients with asthma which closely related to the expression levels of ILC2s and MDSCs associated moleculars. These findings indicated that Th2 polarization was close related to synergistically increased ILC2s and MDSCs, it may allow to further the comprehension of the contribution of these cells to the inflammatory response involved in asthma or other respiratory tract inflammatory diseases, such as COPD and RVI, as well as to develop novel therapeutic strategies.

Abstract Macrophage, a highly plastic population, is widely distributed. Macrophage functions are settled and acquired polarization programs in response to microenvironmental signals and involved in many inflammatory disorders, such as experimental autoimmune myocarditis (EAM). Phenotypic and functional changes in macrophage are considered as an important determinant of disease progression and/or regression. Angiotensin II (ANG II), as a powerful proinflammatory factor, plays critical roles in inflammatory diseases and macrophage recruitment. It remains unclear whether ANG II contributed to the functional skewing of cardiac infiltrated monocytes/macrophage and involved in EAM development. Therefore, the present work was to address the above questions. Our data showed that ANG II contributed to CD11b+Ly6Chi (CD11b+Ly6G−Ly6C+) cells reprogramming into M1-like macrophage through Erk1/2 or p38/Stat3 pathway and the reprogramming M1-like cells promoted Th17 cells expansion; abrogation of ANG II-AT1R axis significantly ameliorated cardiac injury. The present work first demonstrated a novel immune regulation role of ANG II; ANG II, as a powerful immune factor, promoted CD11b+Ly6Chi inflammatory cells reprogramming into M1-like macrophage and involved in inflammatory disorders development; our results also indicated that ANG II may be a potential therapeutic target for inflammatory diseases.

High-mobility group box 1 (HMGB1), an important inflammatory factor, plays significant roles in CD4+T cell differentiation, cancer and autoimmune disease development. Our previous data have demonstrated that HMGB1 contributes to macrophage reprogramming and is involved in experimental autoimmune myocarditis (EAM) development. In contrast to the well-explored function of HMGB1, little is known about the nuclear function. Whether HMGB1 can serve as an architectural factor and control functional skewing of macrophages remains unclear. Therefore, the present work was performed to address the above speculation. The adenovirus-mediated shRNA (Ad-shRNA) was employed to knock down HMGB1 in RAW264.7 and monocytes/macrophages of EAM mice. Our data showed that in vitro HMGB1 silencing limited functional skewing of macrophages and down-regulated inflammatory factors secretion, which can't be reversed by the exogenous HMGB1. In M1 polarization system, the phosphorylations of NF-κB, p38 and Erk1/2 were inhibited following HMGB1 silencing. In vivo, HMGB1 silencing could effectively ameliorate EAM development. Our data suggest that HMGB1 may be a checkpoint nuclear factor of macrophage reprogramming. Our findings also provide an exciting therapeutic method for inflammatory disorders.

As a common nosocomial infection bacterium, A. baumannii’s drug resistance rate continues to rise. In this study, the objective was to explore the possible reasons for the increased drug resistance of A. baumannii after tigecycline treatment. Based on the drug resistance analysis of 183 clinical isolates of A. baumannii, a pair of strains (AB711 and AB721) which changed their resistance after treatment was selected. Tigecycline was used to induce the drug resistance of strain AB711 in vitro. The differential expressed genes from A. baumannii strains were analyzed using whole gene sequencing (WGS) and RNA sequencing (RNA-seq) combined with online MLST, SNP tools and bioinformatics software, and verified by reverse transcription quantitative polymerase chain reaction (RT-qPCR). AB721 became more resistant to tetracyclines than AB711 at the initial detection. However, after a period of time, the resistance of AB711 and AB721 became consistent. This phenomenon can also be repeated using AB711 in vitro. After induction, the AB711 with increased MIC value of tigecycline was named AB712. The results of WGS, MLST and SNP based Phylogenetic tree indicated that AB711, AB712, AB721 were co-origin and belong to ST2 (Pasteur) / ST1791 (Oxford). Comparative transcriptome indicated that the Differential expression of some genes can play an important role in the resistance enhancement process of AB711. For example, compared with AB711, genes related to benzene-containing compound metabolic process, translation, ribosomal structure and biogenesis and so on were upregulated significantly in AB712. In addition, efflux pumps such as RND transporter permease subunit, EmrAB, MacB, and Tet resistance operon were also upregulated. Tigcycline induced changes in the expression of some related genes in A. baumannii, which may be the main reason for its increased drug resistance.

Porphyromonas gingivalis is regarded as one of the risk factors of periodontitis. P. gingivalis exhibits a wide variety of genotypes. Many insertion sequences (ISs), located in their chromosomes, made P. gingivalis differentiate into virulent and avirulent strains. In this research, we investigated the prevalence of P. gingivalis in the gingival crevicular fluid (GCF) among periodontitis patients from Zhenjiang, China, detected the P. gingivalis rag locus distributions by multiplex polymerase chain reaction (PCR), and analyzed the origin of the P. gingivalis rag locus based on evolution. There were three rag locus variants co-existing in Zhenjiang. The results showed that the rag locus may be associated with severe periodontitis. This work also firstly ascertained that the rag locus might arise, in theory, from Bacteroides sp. via horizontal gene transfer.

Corynebacterium pyruviciproducens is a newly discovered Corynebacterium species with no known pathogenic components such as diphtheria toxin and tuberculostearic acid, and it has similar biological properties to Propionibacterium acnes, but its role of immunoregulation is drawing people's attention. In this work, based on the role of macrophages in removal of pathogenic bacteria as a primary scavenger and particulate antigen-presenting cell, the stimulation of macrophages by C. pyruviciproducens was analyzed through detecting the levels of cytokine secretion and expression of membrane molecules, and the effect of C. pyruviciproducens in promoting antibody response to sheep red blood cells (SRBC) in vivo was detected. In vitro, C. pyruviciproducens led to a sharp release of interleukin-6 and tumour necrosis factor-α and encouraged the activation of macrophages including enhanced expressions of MHC-II, CD40, CD80 and CD86. In vivo, it enhanced the humoral immune response against SRBC, a particulate antigen. These observations suggest that C. pyruviciproducens, as an immunoregulator, can promote the host humoral immune response to pathogenic microorganisms by regulating macrophage function.

Enterovirus 71 (EV71) is the most important etiological agent of hand, foot, and mouth disease (HFMD) in young children, which is associated with severe neurological complications and has caused significant mortalities in recent HFMD outbreaks in Asia. However, there is no effective antiviral therapy against EV71. In this study, RNA interference (RNAi) was used as an antiviral strategy to inhibit EV71 replication. Three small interfering RNAs (siRNAs) targeting the 2Apro region of the EV71 genome were designed and synthesized. All the siRNAs were transfected individually into rhabdomyosarcoma (RD) cells, which were then infected with strain EV71-2006-52-9. The cytopathic effects (CPEs) in the infected RD cells, cell viability, viral titer, and viral RNA and protein expression were examined to evaluate the specific viral inhibition by the siRNAs. The results of cytopathogenicity and MTT tests indicated that the RD cells transfected with the three siRNAs showed slight CPEs and significantly high viability. The 50% tissue culture infective dose (TCID50) values demonstrated that the viral titer of the groups treated with three siRNAs were lower than those of the control groups. qRT–PCR and western blotting revealed that the levels of viral RNA and protein in the RD cells treated with the three siRNAs were lower than those in the controls. When RD cells transfected with siRNAs were also infected with strain EV71-2008-43-16, the expression of the VP1 protein was significantly inhibited. The levels of interferon α (IFN-α) and IFN-β did not differ significantly in any group. These results suggest that siRNAs targeting the 2Apro region of the EV71 genome exerted antiviral effects in vitro.

Glucocorticoid‑induced tumor necrosis factor receptor (GITR) is expressed at high levels on CD4+CD25+ regulatory T cells (Tregs). Following activation by its ligand (GITRL), GITR influences the activity of effector T cells and Tregs and participates in the development of numerous autoimmune and inflammatory diseases, including asthma. However, the GITR/GITRL expression level in lung tissue and its influence on group 2 innate lymphocytes (ILC2s) in asthma remains unclear. The present study detected the number of ILC2s and the expression levels of GITR and GITRL in the lung tissues of asthmatic mice by flow cytometry analysis, immunofluorescence staining and reverse transcription quantitative polymerase chain reaction. The results demonstrated that the number of ILC2s and the expression levels of ILC2‑associated molecules (interleukin‑33 receptor ST2, RAR related orphan receptor A and inducible T cell costimulator) were increased in the lung tissues of asthmatic mice. The upregulated ILC2s were accompanied by an increased number of GITR‑positive cells in the spleen and lung tissues, and additionally an increased level of GITRL mRNA in lung tissue in asthma. In addition, increased mRNA expression levels of interleukin (IL)‑5 and IL‑13 were observed in the asthmatic lung, and there was a significant, positive correlation between the mRNA levels of GITR/GITRL and ILC2‑associated molecules. Therefore, GITRL treatment may increase the number of ILC2s and/or GITR‑positive cells in lung tissue. These results indicated that the activity of GITR‑expressing ILC2s may be enhanced via interaction of GITRL and GITR, which may contribute to pathogenesis of asthma. These findings present potential therapeutic targets for the treatment of asthma.

Cancer stem cells (CSCs) or cancer-initiating cells (CICs) play an important role in tumor initiation, progression, metastasis, chemoresistance, and recurrence. It is important to construct an effective method to identify and isolate CSCs for biotherapy of cancer. During the past years, many researchers had paid more attention to it; however, this method was still on seeking. Therefore, compared to the former methods that were used to isolate the cancer stem cell, in the present study, we tried to use modified transwell system to isolate and enrich CSCs from human pancreatic cancer cell lines (Panc-1). Our results clearly showed that the lower chamber cells in modified transwell system were easily forming spheres; furthermore, these spheres expressed high levels of stem cell markers (CD133/CD44/CD24/Oct-4/ESA) and exhibited chemoresistance, underwent epithelial-to-mesenchymal transition (EMT), and possessed the properties of self-renewal in vitro and tumorigenicity in vivo . Therefore, we speculated that modified transwell assay system, as a rapid and effective method, can be used to isolate and enrich CSCs.

ABSTRACT Angiotensin II (ANG II) plays critical roles in modulation of circulatory homeostasis and activation of innate and adaptive immunity and has also been implicated in several mouse models of autoimmune disease. However, how ANG II regulates macrophages and is involved in development of experimental autoimmune myocarditis (EAM) remains unclear. Therefore, the present study aimed to address the above question and explore possible mechanisms. EAM was induced in BALB/c mice. ANG II was quantitated by ELISA and hematoxylin and eosin staining was employed to analyze pathological changes and macrophage infiltration. The chemotactic ability of ANG II was assessed by using a Transwell system. It was found that ANG II is up‐regulated in serum and heart tissues of mice with EAM and that ANG II significantly drives monocyte/macrophage infiltration through the C‐C chemokine receptor 2/5 (CCR2/5) axis. CCR2/5 antagonists and ANG II receptor inhibitor could all abrogate monocyte/macrophage infiltration and ameliorate development of EAM. Our results have firstly identified a novel function of ANG II: that it is a critical chemokine for monocyte/macrophage recruitment. Furthermore, our results indicate that ANG II is a potential candidate for treatment of inflammatory diseases.

Abstract The accumulation of airway apoptotic cells may be an important factor causing airway hyper‐responsiveness (AHR). Whether the apoptotic cells can be promptly removed is related to the occurrence and course of asthma. In recent years, studies have shown that Rac1 is involved in many cellular biological activities including the formation and elimination of apoptotic cells. In this study, based on the analysis of airway local cells and related factors in asthmatic mice, we evaluated the expression of Rac1 in airway epithelial cells or phagocytes and analysed its relationship with the incidence of apoptosis or scavenging of apoptotic cells. Our data showed that the expression level of Rac1 in asthmatic mice decreased significantly, while the expression of IL ‐33 increased obviously. The airway epithelial cell line was stimulated by curcumin at 50 μmol/L for 24‐48 hours; more than 50% of the cells were apoptotic, and of which, about 20% were late apoptosis. Rac1 inhibitor ( NSC 23766) can enhance the apoptosis effect. In addition, the ability of phagocytosis and migration in the epithelial cells or macrophages was increased following the application of Rac1 inhibitors or specific si RNA in a dose‐dependent manner, and the expression level of IL ‐33 was simultaneously increased after blocking Rac1. It is suggested that the down regulation of Rac1 in asthma may contribute to the apoptosis of airway epithelial cells and affect the clearance of apoptotic cells, which will lead to the aggregation of the apoptotic cells in the respiratory tract and participate in AHR.

Corynebacterium pyruviciproducens (C. pyruviciproducens), a newly discovered Corynebacterium, is gram-positive, non-flagellate, non-spore-forming lipophilic rod. No known pathogenic components of Corynebacteria have been found in this new bacterium, such as diphtheria toxin and tuberculostearic acid. In the present study, referring to Propionibacterium acnes (P. acnes), a well-known bacterial adjuvant, the stimulation of dendritic cells by C. pyruviciproducens was analyzed through detecting the levels of cytokine-secretion, ability of cell-proliferation and expression of membrane molecules. In addition, the effect of C. pyruviciproducens in promoting antibody production in vivo was detected. Compared with P. acnes, C. pyruviciproducens more strongly enhanced cytokine secretion including inflammatory factor IL-6 and Th1-associated molecule IL-12, and more effectively induced proliferation, activation or maturation of D2SC/1 (a murine dendritic cell line) and bone marrow-derived dendritic cells (BMDC). Vaccination studies in mice using ovalbumin (OVA) as a model antigen showed that C. pyruviciproducens effectively promoted antigen-specific humoral immune response by increasing OVA-specific antibody, Th2-biased response in spleen and high IL-4/IFN-γ ratio in serum.

The outer membrane protein RagB is one of the major virulence factors of Porphyromonas gingivalis (P. gingivalis). To prevent periodontitis and associated systemic diseases induced by P. gingivalis, we built B cell antigen epitope vaccine characterized by pIRES-ragB'-mGITRL to induce a protective immune responses. The B cell antigen epitope and scrambled peptide of ragB were predicted, cloned into pIRES and constructed pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. pIRES-ragB'-mGITRL was transfected into COS-7 cells. Subsequently, the 6-week-old female BALB/c mice were challenged by P. gingivalis following three time immunization by pIRES, pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. The levels of RagB-specific antibody in the serum and Tfh cells in the spleen were measured by ELISA and flow cytometry, respectively. And higher levels of RagB-specific IgG were produced in the immunized mice with pIRES-ragB'-mGITRL. Additionally, the number of Tfh cells was also expanded and lesions were diminished in pIRES-ragB'-mGITRL mice comparing with control groups. Our results clearly demonstrated that P. gingivalis B cell antigen epitope vaccine, pIRES-ragB'-mGITRL, could induce protective immune responses. Furthermore, our data also indicated that pIRES-ragB'-mGITRL was a potential therapeutic vaccine against P. gingivalis.

At present, collagen‑induced arthritis (CIA) is the best known and most extensively used model for the immunological and pathological characteristics of human rheumatoid arthritis (RA). This model is useful not only in aiding our understanding of the pathogenesis of this disease, but also in the development of new therapies. Bovine, porcine and human collagen has been used to induce CIA; however, response has been identified to vary between strains and injection conditions, and false positive results and reduced potency are common as a result of minor contaminants or deglycosylated protein. Therefore, in the present study, type II collagen (CII) was isolated and purified from chicken sternal cartilage and was found to successfully induce the RA model. Furthermore, T helper 17 (Th17) cells were observed to infiltrate the joint on day 45 following induction by CII. In vitro, expression of toll‑like receptor 2 (TLR2) increased in peritoneal macrophages stimulated by CII. In addition, blockage of TLR2 was identified to markedly decrease levels of TGF‑β and IL‑6 in the cell culture supernatant. The results indicate that CII isolated from chicken sternal cartilage may be recognized by TLR2 on macrophages, leading to TGF‑β and IL‑6 production and subsequent activation of Th17 cells which mediates CIA development.

Acinetobacter baumannii (A. baumannii), especially the multidrug resistant A. baumannii (MDR-AB) is becoming a common opportunistic pathogen in hospital, and constitutes significant public health threats. This study aimed at investigating the relationship between drug resistance with expression of class A-D β-lactamase genes, mutation in membrane porin and over-expression of efflux pump genes among A. baumannii isolated from Zhengjiang, China.Antibiotic susceptibility assays were performed using Kirby-Bauer disc diffusion method. PCR was used to detect β-lactamase genes and carO, oprD, adeR, adeS. Real-time PCR was used to assess the mRNA expression level of efflux pump gene adeB. The software of DNAMAN was applied to assemble oprD and carO sequences, and the sequences were compared with those retrieved from GenBank (http://www.ncbi.nlm.nih.gov/).27 isolates (61.4%) in this study were MDR-AB, in which five β-lactamases including TEM, CTX-M-2, ADC, OXA-23 and OXA-51 were found, and the positive rate was 96.3% (26), 14.8% (4), 92.6% (25), 88.9% (24) and 92.6% (25), respectively. In addition, the expression level of adeB mRNA was significantly increased in MDR-AB, it might due to adeR mutation. Some mutations were also found in carO and oprD.MDR-AB showed high relationship with β-lactamase, mutation in membrane porin and overexpression of adeB, which may directly relates to the mutation in regulating gene adeR.

Abstract Type 2 innate lymphoid cells (ILC2s) have multiple functions that can respond to allergic diseases, parasite infection, metabolic homeostasis, tissue repair, and adipose metabolism homeostasis. In these diseases, ILC2s can be activated by various inflammatory cytokines released by damaged cells. Activated ILC2s produce different type 2 cytokines, including interleukin (IL)‐4, IL‐5, IL‐9, and IL‐13, which involved in the pathogenesis of many diseases. In recent years, the relationship between ILC2s and tumor diseases has attracted more and more attention. The role of ILC2s in tumor immunity depends on its surface molecules and cytokine context. This review aims to conclude tumorigenic and antitumorigenic roles of ILC2s, and the characters of ILC2s‐related cytokines in tumor diseases to provide a comprehensive overview of the impact of ILC2s in tumor immunity.

Th1 and Th17 cells are involved in the pathogenesis of rheumatoid arthritis (RA). T-bet, a Th1-specific transcription factor, appears to drive the maturation of Th1 and IFN-γ secretion. In the present study, we established the T-bet shRNA recombinant plasmid (p-T-shRNA) and explored its possible anti-inflammatory effect in a collagen-induced arthritis (CIA) model by local injection of plasmid vectors. For the initiation of CIA, DBA/1J mice were immunized with type II collagen (CII) in Freund's adjuvant and the CII-immunized mice were treated with p-T-shRNA. Levels of T-bet, IFN-γ, IL-17 and RORγt mRNA in splenocytes and synovial joints were measured by quantitative real-time PCR and T-bet expression in joint tissue was detected by immunohistochemistry staining. The intracellular IFN-γ and IL-17 were analyzed by flow cytometry (FCM). The results demonstrated that therapeutic administration on the local joints with p-T-shRNA significantly suppressed IFN-γ and IL-17 gene expression and improved the pathogenesis of arthritis in CIA mice, while administration of a plasmid expressing T-bet (pIRES-T-bet) accelerated the disease onset. Our study suggests that T-bet may be developed as a potential target for arthritis therapy.

Macrophage plays a critical part in host defense, tissue repair, and anti-inflammation; Macrophage reprogramming is responsible for disease development or regression. We aimed to clarify the effect of sinomenine-4-hydroxy-palmitate (C16), on macrophage reprogramming and anti-inflammatory in endotoxemia model. According to a structure modification of SIN (Sinomenine), C16 was found. Then, based on the endotoxin model, the mice liver and kidney toxicity was evaluated and serum cytokines level of IL-6 (Interleukin-6), TNF-α (Tumor necrosis factor-α), and IL-1β (Interleukin-1β) were measured by ELISA (Enzyme linked immunosorbent assay). Then, we confirmed the effect of C16 on macrophages reprogramming, we used the flow cytometry to test the effect of C16 on macrophages apoptosis in vitro. Then, iNOS (Inducible nitric oxide synthase), M1-type related cytokines, such as IL-1β, TNF-α, and M2-type related cytokines, such as Arg-1 (Arginase-1), CD206, Fizz1, and Ym1 was detected, which expressed in ANA-1 and primary peritoneal macrophages. To further explore the molecular mechanism of C16 in reprogramming of macrophages from M1 toward M2 phenotype, the expression of STAT1 (signal transducer and activator of Transcription 1), STAT3, ERK1/2 (extracellular signal regulated kinase1/2), AKT, p38, and its corresponding phosphorylation were determined by western blot. Our results demonstrated that C16 improved the survival rate of LPS- (lipopolysaccharide) challenged mice and decreased the inflammatory cytokines expression; After C16 treatment, the expression of M1 phenotype correlation factors decreased significantly, while the expression of M2 phenotype correlation factors increased significantly at different levels compared with normal group. It indicated that C16 reprogram macrophages phenotype from M1 toward M2 following LPS stimulus. Furthermore, the results also showed that C16 showed anti-inflammatory effect by inhibiting LPS-induced p38, AKT and STAT1 phosphorylation and contributing ERK1/2 activation. C16 promoted macrophage reprogramming toward M2-like phenotype via p-p38/p-AKT or STAT1 signals pathway and C16 might be a valid candidate for inflammatory disease.

A bioflocculant-producing bacterium named C-2 was isolated from soil.The strain was identified as Pseudomonas fluorescens by nPCR amplification kit,and flocculating experiment showed that in alkali condition and at the presence of CaCl2,the bioflocculant named PS-2 had high efficiency with small dosage for dealing with kaolin suspension with larger floc,rapid sedimentation rate over a wide range of temperature.The bioflocculant had good flocculation activity to many kinds of wastewater.Component analysis showed that PS-2 was composed of polysaccharide and nucleic acid.

As a distinctive population of leucocytes, innate lymphoid cells (ILCs) participate in immune-mediated diseases and play crucial roles in tissue remodelling after injury. ILC lineages can be divided into helper ILCs and cytotoxic ILCs. Most helper ILCs are integrated into the fabric of tissues and produce different types of cytokines involving in the pathogenesis of many kinds of cardiovascular disease and form intricate response circuits with adaptive immune cells. However, the specific phenotype and function of helper ILC subsets in cardiovascular diseases are still poorly understood. In this review, we firstly highlight the distribution of helper ILCs in cardiovascular system and further discuss the potential contribution of helper ILCs in inflammation-associated cardiovascular disease.

Newly identified nuocytes play an important role in Th2 cell mediated immunity such as protective immune responses to helminth parasites, allergic asthma and chronic rhinosinusitis. However, the contributions of nuocytes in the occurrence and development of Graves' hyperthyroidism remains unknown. Previous studies found that there was a predominant Th2 phenotype in patients with Graves' hyperthyroidism, it might relate to polarization of nuocytes. Nuocytes were defined by transcription factor RORα, various cell surface markers (T1/ST2, IL-17RB, ICOS, CD45) and associated cytokines. In this study, these cells related genes or molecules in PBMC from patients with Graves' hyperthyroidism were measured, and the potential correlation between them was analyzed. The expression levels of T1/ST2, IL-17RB, ICOS, IL-5 and IL-13, which represented nuocytes associated molecules were significantly increased in patients, meanwhile, the RORα mRNA also had a tendency to increase. In addition, IFN-γ and T-bet (Th1 related cytokine and transcription factor) were obviously decreased, and there was a positive correlation between IL-17RB and IL-13. These results suggested that there were polarized nuocytes in Graves' hyperthyroidism patients, and which closely related to the down-regulation of Th1 cells or relatively advantage of Th2 differentiation.

To synthesize the derivatives of sinomenine, 4-palmitoyl-sinomenine, and explore its therapeutic effect on lipopolysaccharide (LPS)-induced endotoxemia.A highly efficient synthesis of sinomenine derivatives called 4-palmitoyl-sinomenine was made with a molecule of palmitic acid substitutions at C-16 position of ring A. One hour before endotoxemia induction by i.p. injection of 10 mg/kg LPS, high-dose treatment mice (n=5/group) received an i.p. injection of 5 mg/kg sinomenine or 4-palmitoyl-sinomenine while the low-dose treatment mice (n=5/group) received 2.5 mg/kg sinomenine or 4-palmitoyl-sinomenine. Untreated group and normal control group received normal saline. And their survival was monitored hourly for 24 hours. Examination of cytotoxicity of 4-palmitoyl-sinomenine on RAW264.7 cells was conducted at a concentration range of 1 to 125 μmol/L using MTT assay. RAW264.7 cells were exposed to 4-palmitoyl-sinomenine (0, 1, 2, 5, 10 μmol/L) for 24 hours, and then treated with LPS (1 μg/mL) for 6 hours. Then RAW264.7 cells were collected and the mRNA level of IL-6 was detected by real-time quantitative PCR(qRT-PCR).Sinomenine derivatives were successfully synthesized to get 4-palmitoyl-sinomenine. The survival percentage of 4-palmitoyl-sinomenine treatment groups was higher than that of sinomenine treatment groups at the same treatment concentration. The 4-palmitoyl-sinomenine inhibited RAW264.7 cell proliferation and IL-6 gene transcription.The 4-palmitoyl-sinomenine has an anti-inflammation probably through inhibiting the proliferation of RAW264.7 cells and decreasing the inflammatory gene expression and inflammatory cytokine release.

Macrophages play critical roles in inflammatory initiation, development, resolution and cardiac regeneration of myocarditis. However, Reg3β, as a member of regenerating family of proteins, contributes to dedifferentiation of injury cardiomyocytes as well as cardiac function remodeling. It remains unclear whether Reg3β was associated with macrophages reprogramming during autoimmune myocarditis. Our results showed that Reg3β could effectively recruit macrophages, promoted their proliferation and phagocytosis, and facilitated their polarized into M2 macrophages. Macrophage, especially M1 phenotype contributed to Reg3β production by cardiomyocytes. Our data also indicated that Reg3β was involved in self-protection mechanism following cardiac injury or stress. This suggests that Reg3β might be a critically protective factor of myocardium.

Since regenerating islet-derived 3β (Reg3β) was first reported, various studies have been conducted to explore the involvement of Reg3β in a gamut of maladies, such as diabetes, pancreatitis, pancreatic ductal adenocarcinoma, and extrapancreatic maladies such as inflammatory bowel disease, acute liver failure, and myocardial infarction. Surprisingly, there is currently no systematic review of Reg3β. Therefore, we summarize the structural characteristics, transcriptional regulation, putative receptors, and signaling pathways of Reg3β. The exact functional roles in various diseases, especially gastrointestinal and liver diseases, are also discussed. Reg3β plays multiple roles in promoting proliferation, inducing differentiation, preventing apoptosis, and resisting bacteria. The present review may provide new directions for the diagnosis and treatment of gastrointestinal, liver, and pancreatic diseases.

Glucocorticoid‑induced tumor necrosis factor receptor related protein (GITR) is a member of the tumor necrosis factor receptor superfamily. The present study attempted to obtain the mouse GITR‑Fc fusion protein and investigate its function on the proliferation of CD4+ T cells and on the expression of mGITR ligand (mGITRL) on dendritic cells. The sequences of the mouse (m)GITR gene and mouse immunoglobulin G Fc (mIgGFc) were amplified from mouse spleen cells and introduced into a pET‑32a(+) vector. Following the induction, purification and validation of the mGITR‑Fc fusion protein, the mGITR‑Fc fusion protein was used to analyze its function on the proliferation of CD4+ T cells and on the expression of mGITR on dendritic cells. A recombinant plasmid containing the mGITR gene fragment and mIgGFc was constructed, and the recombinant mGITR‑Fc fusion protein was successfully expressed. The exogenous mGITR‑Fc fusion protein inhibited the proliferation of CD4+ T cells, dependent on the presence of mGITRL. The exogenous mGITR‑Fc fusion protein also inhibited the expression of mGITRL on the dendritic cells. In conclusion, the mGITR‑Fc fusion protein was confirmed to exhibit biological functions of a co‑stimulatory signal and reverse signal. These experiments provide the basis for further investigation of the function of the mGITR‑Fc fusion protein on certain autoimmune diseases.

To detect the levels of miR-21a-5p, miR-155-5p, miR-218-5p, miR-222-3p, miR-494-3p in ovarian cancer tissues and the number of myeloid-derived suppressor cells (MDSCs) in peripheral blood and spleen in ovarian cancer-bearing mice, and explore their clinical significance and correlations.The mRNA expressions of miR-21a-5p, miR-155a-5p, miR-218-5p, miR-222-3p, miR-494-3p were detected by real-time quantitative PCR (qRT-PCR) in tumor tissues and tumor-adjacent normal tissues from 12 ovarian cancer-bearing mice. The frequency of MDSCs in the peripheral blood and spleen from the 12 tumor-bearing mice was measured by flow cytometry. Spearman correlation analysis was used to find out the correlations between MDSCs and miRNAs.Compared with the normal mice, the number of MDSCs in the peripheral blood and spleen of the tumor-bearing mice significantly increased. The levels of miR-21a-5p, miR-218-5p and miR-222-3p in tumor tissues were significantly higher than those in tumor-adjacent normal tissues; conversely, miR-155a-5p and miR-494-3p levels in the former were significantly lower than those in the latter. There was no correlation between miR-222-3p and MDSCs, but miR-494-3p had a negative correlation with MDSCs. The expression differences of miR-21a-5p, miR-155a-5p, miR-218-5p between tumor tissues and tumor-adjacent normal tissues had positive correlations with the number of MDSCs.The levels of miR-21a-5p, miR-155a-5p, miR-218-5p, miR-494-3p in tumor tissues had correlations with the number of MDSCs in the peripheral blood and spleen of ovarian cancer-bearing mice.

The recent identification of a complex group of innate lymphocyte cells, now collectively termed innate lymphoid cells (ILCs), has been implicated in the pathogenesis of inflammatory disorders. Basically, three classes of ILCs have been described according to how they function and develop as well as the type of cytokines they produce and how the interactions between innate and adaptive immune cells are influenced during inflammatory disorders. Data from clinical and experimental animal models suggest that ILCs modulate various autoimmune diseases. The ILCs play this role possibly through their ability to produce cytokines, which influence the activities of important cells, particularly the effector CD4+ cells. Due to their identified functions in the pathogenesis of autoimmune diseases, researchers have proposed that they may be viable therapeutic targets. This review outlines the biology and functions of ILCs in autoimmune diseases and attempts to further identify their potential as therapeutic targets.

Objective To detect the expression levels of transcription factors and associated cytokines of Th17 and Treg cells in peripheral blood mononuclear cells （ PBMC ） of patients with gastric cancer, and explore the possible pathological mechanism of these cells involved in the development of gastric cancer. Methods The mRNA levels of RORγt, FoxP3 in PBMC were determined by quantitative real-time PCR （ QRT-PCR ） from 57 patients with gastric cancer, 31 patients with benign gastric illness and 40 healthy people. The concentration of IL-17, IL-23, TGF-β, IL-10 in plasma were detected by enzyme linked immunosorbent assay （ELISA）. Results Compared with healthy volunteers, patients with gastric cancer showed higher levels of RORγ/t and FoxP3 in PBMC （ P 〈 0.05 ）. The ratio of FoxP3/RORγ/t in gastric cancer group was higher than that in the volunteer group and benign gastric illness group （ P 〈 0.05 ）. The ratio of FoxP3/RORγ/t was higher in advanced disease than early disease （ P 〈 0.05 ）. The expressions of IL- 17, IL-23, TGF-β and IL-10 were higher in patients with gastric cancer than that in healthy volunteers （P 〈 0.05）. In addition, The expression of TGF-β and IL-10 were significantly increased in the advanced disease group than that in the early group （P 〈0.05） , but IL-17 and IL-23 was not significantly changed between the two groups （P 〉 0. 05）. Conclusion There are higher levels of Th17 and Treg cells in gastric cancer patients, and it also shows a persistent predominant tendency of Treg cells and a reduced tendency of Th17 cells in advanced disease. Detecting the expression of Th17/Treg transcription factor and related cytokines would contribute to the diagnosis and prediction of the disease development and prognosis. Key words: Gastric neoplasms ;  RORγt ;  FoxP3 ;  Th17 cell ;  Treg cell

Hlx as a Th1-specific transcription factor, it appears to drive maturation of Th1 and IFN-γ secretion in cooperation with T-bet. In this study, we established a stable Hlx-over-expressed dendritic cell line (DC2.4/Hlx), and investigated the possible effect of Hlx gene on maturation of dendritic cell-line (DC2.4). Results shown that over-expressed Hlx in DC2.4 up-regulated the transcription and expression of IFN-γ, increased the expression of maturation makers including CD40, CD80, CD86, MHC-I and MHC-II. Functional assay for DC2.4/Hlx showed that over-expressed Hlx increased the expression level of interleukin-12 in the supernatant and decreased DC endocytosis when cells were incubated in vitro. Furthermore, using a syngeneic T cell activation model, we found that DC2.4/Hlx could obviously present ovalbumin (OVA) antigen to T cell in OVA pre-immunized mice.

OBJECTIVE To explore the relationship between resistance of multidrug-resistant Acinetobacter baumannii and transposon and insertion sequence.METHODS K-B method was used to detect the resistance.PCR was used to analyze the transposon and insertion sequence.RESULTS The resistant rates of A.baumannii were cefotaxime 78.9%,ceftazidime 63.1%,cefepime 78.9%,ceftazidime 100.0%,imipenem 73.7%,meropenem 73.7%,ampicillin 78.9%,amoxicillin 78.9%,amikacin 52.6%,gentamicin 57.8%,ciprofloxacin 78.9% and tetraycline 78.9%,respectively.The detection rate of tnp513 was 47.4%,Isaba1 was 78.9% and IS 26 was 47.4%.CONCLUSION The resistance of multi-drug resistant A.baumannii is associated with transposon and insertion sequence.

Abstract Background Acinetobacter baumannii is a multidrug resistant (MDR) hazardous bacterium with very high antimicrobial resistance profiles. Outer membrane vesicles (OMVs) help directly and/or indirectly towards antibiotic resistance in these organisms. The present study aims to look on the proteomic profile of OMV as well as on the bacterial transcriptome upon exposure and induction with eravacycline, a new synthetic fluorocycline. RNA sequencing analysis of whole cell and LC–MS/MS proteomic profiling of OMV proteome abundance were done to identify the differential expression among the eravacycline-induced A. baumannii ATCC 19606 and A. baumannii clinical strain JU0126.Results The Differential expressed genes from the RNA sequencing and were analysed using R package and bioinformatics software and tools. Genes encoding drug efflux and membrane transport were upregulated among the DEGs from both ATCC 19606 and JU0126 strains. As evident with the induction of eravacycline resistance, ribosomal proteins were upregulated in both the strains in the transcriptome profiles and also resistance pumps, such as MFS, RND, MATE and ABC transporters. High expression of stress and survival proteins were predominant in the OMVs proteome with ribosomal proteins, chaperons, OMPs OmpA, Omp38 upregulated in ATCC 19606 strain and ribosomal proteins, toluene tolerance protein, siderophore receptor and peptidases in the JU0126 strain. The induction of resistance to eravacycline was supported by the presence of upregulation of ribosomal proteins, resistance conferring factors and stress proteins in both the strains of A. baumannii ATCC 19606 and JU0126, with the whole-cell gene transcriptome towards both resistance and stress genes while the OMVs proteome enriched more with survival proteins.Conclusion The induction of resistance to eravacycline in the strains were evident with the increased expression of ribosomal and transcription related genes/proteins. Apart from this resistance conferring efflux pumps, outer membrane proteins and stress related proteins were also an essential part of the upregulated DEGs. However, the expression profiles of OMVs proteome in the study was independent with respect to the whole cell RNA expression profiles with low to no correlation. This indicates the possible role of OMVs to be more of back-up additional protection to the existing bacterial cell defence during the antibacterial stress.

Dear Editor, Recently, we demonstrated that lncRNA187415.1 knockdown in breast cancer-associated macrophages (BCAMs) could change their phenotype and function of BCAMs, benefit the BCAMs inducing breast cancer cells apoptosis and inhibit their invasion. LncRNA187415.1 maintained BCAMs function in local microenvironment by inhibiting CISH mRNA degradation; however, transcription factor C/EBPβ regulated transcription of lncRNA187415.1 via binding promoter region of –1500∼–2000 bp of lncRNA187415.1. As our previous research about lncRNA profiles of BCAMs using lncRNAs microarray showed that1 lncRNA187415.1 was the most significantly changed gene. The data were further confirmed in BCAMs isolated from breast cancer bearing mice in vitro (Figures 1A and 1B). To further investigate the biological role of lncRNA187415.1, its expression in BCAMs was successfully silenced by siRNAs (Figure 1C). LncRNA187415.1 silence in BCAMs significantly inhibited the proliferation, migration, and invasion of 4T1 cells (Figures 1D and 1E). Conversely, lncRNA187415.1 silence in BCAMs resulted in a significantly increase of the apoptosis in 4T1 cells (Figure 1F). To further confirm the similar effects also exist in vivo, the jetPEI-Man/ si-lncRNA187415.1 complex was used to specifically downregulate the lncRNA187415.1 in BCAMs of breast cancer bearing mice. A dramatic reduction of tumor volumes and tumor weight were observed in si-lncRNA187415.1 group compared to si-NC (Figures 1G and 1H). These results clearly demonstrated that lncRNA187415.1 knockdown target on BCAMs significantly ameliorated breast cancer progression. LncRNA187415.1 silence in BCAMs ameliorated breast cancer progression. (A) Volcano plot of lncRNAs gene profiles results in BCAMs. The expression of lncRNAs in macrophages and BCAMs was detected by lncRNAs microarray technology. Upregulation (red), downregulation (green), and no-significance (gray). (B) qPCR analysis for lncRNA187415.1 expression in BCAMs from breast tumors (n = 8). (C) siRNA knockdown efficiency. siRNAs (50 nM) were transiently transfected into BCAMs, after 48 h, BCAMs were harvested for qPCR analysis (n = 3). (D) Representative images of 4T1 are stained by Ki67 (green) and DAPI (blue). LncRNA187415.1-downregulated BCAMs were co-cultured with 4T1 for 48 h. The proliferation of 4T1 was evaluated by Ki-67/DAPI staining. Data were statistically analyzed based on three independent experiments. (E) Invasion and migration ability of 4T1 cells co-cultured with lncRNA187415.1 downregulated BCAMs. Data represent means ± SD from three independent experiments. (F) Apoptosis of 4T1 cells was analyzed by Annexin V and PI staining. Note that 4T1 cells were co-cultured with LncRNA187415.1-downregulated BCAMs for 48 h; the apoptosis of 4T1 cells was analyzed by flow cytometry (n = 3). (G) Tumor volume. Note that 4T1 tumor-bearing mice were given jetPEI-Man/si-NC complex or jetPEI-Man/siRNA2 complex. Tumor volume measurement was performed by digital caliper at the indicated days (n = 7). (H) Tumor weight. Tumors were resected at day 24, and tumor weight was measured (n = 7). All data are shown as means ± SD. **p < 0.01, ***p < 0.01 Abbreviation: PM, peritoneal macrophage. To determine the effect of lncRNA187415.1 on the phenotype and function of BCAMs, lncRNA187415.1 was silenced in BCAMs with siRNA. After lncRNA187415.1 silence, CD86 and MHC II were significantly increased, conversely, CD206 was downregulated, LPS-treated macrophages (pro-inflammatory type) and IL-4-treated macrophages (anti-inflammatory type) were used as positive or negative control, respectively (Figure 2A). The qPCR and ELISA also indicated that the expression of Tnfα and Il-1β mRNA as well as their proteins levels were also increased after lncRNA187415.1 silence, and the mRNA and protein expression of Il-4, Il -10 were decreased in si-lncRNA187415.1 group (Figures 2B and 2D). Furthermore, the protein level of Arginase 1 (Arg1) decreased and iNOS increased in si-lncRNA187415.1 group (Figure 3C). Taken together, these results suggested that lncRNA187415.1 silence could reprogram the phenotype of BCAMs. As the results above showed that lncRNA187415.1 silence in BCAMs could benefit the shift of BCAMs phenotype and function from anti-inflammatory to pro-inflammatory. To further clarify the mechanism of lncRNA187415.1 maintaining BCAMs showing anti-inflammatory phenotype in local microenvironment, downstream target genes related to lncRNA187415.1 in BCAMs were analyzed by RNA microarray. The results indicated that CISH is the potential target gene (Log2FC = 8.07) (Figure S1A and Table S1). Previous data also indicate that CISH is related to the polarization of macrophages.2 To confirm whether lncRNA187415.1 could promote the expression of CISH, firstly, lncRNA187415.1 was silence in BCAMs, lncRNA187415.1 silence significantly reduced the expression of CISH mRNA and protein level (Figures 3A and 3B). To further confirm how lncRNA187415.1 regulated CISH, actinomycin D, an RNA polymerase inhibitor, was employed. After actinomycin D treatment, the half-life of CISH mRNA was significantly shortened in lncRNA187415.1 silence group compared with control (Figures 3C and 3D), which indicated that lncRNA187415.1 could be involved in transcription-related processes of CISH and facilitate the stability of CISH mRNA. To further confirm, the shift of BCAMs phenotype and function caused by CISH downregulation induced by lncRNA187415.1 silence, overexpressed plasmid of CISH was transfected into the BCAMs following lncRNA187415.1 silence for 48 h, the results showed that expression of CD86 and MHC II decreased, conversely, CD206 increased (Figure 3E). To clarify which molecule regulates lncRNA187415.1 in BCAMs, PROMO database (http://alggen.lsi.upc.es/home.html) was used to predict the transcription factors that bind to lncRNA187415.1 promoter region. C/EBPβ was a candidate transcription factor for lncRNA187415.1. To further investigate the regulation of C/EBPβ on lncRNA187415.1, Cebpb was silence in BCAMs by siRNA (Figure S1B and S1D), the expression of lncRNA187415.1 and Cish was significantly decreased (Figure S1C and S1E), which confirmed that C/EBPβ regulated the expression of lncRNA187415.1 as a transcript factor. Then the JASPER database (http://jaspar.genereg.net/) was used to predict eight sites that could bind to the lncRNA187415.1 promoter region as follows (Figure S1F), and sequential truncations of this lncRNA187415.1 luciferase reported were created to identify regions critical for lncRNA187415.1 transcription. Four luciferase reporter plasmids containing pro-all (including all the sites), pro-1500 (containing five sites), pro-800 (containing three sites), and pro-500 (containing one site) were constructed in order to determine the approximate binding region. The result indicated that the fewer binding sites included, the lower the luciferase activity was. Additionally, Only the pro-1500 of the transcription factor was activated and bound to more than 30% (C/EBPβ+Lnc-pro-luc/C/EBPβ EV+Lnc-pro-luc) (Figures 3F and 3G); therefore, the C/EBPβ bound to the lncRNA187415.1 promoter region from –1.5 kb to –2 kb, and ChIP-qPCR experiment was performed to verify the binding sites (Figure S1G). Our data clearly demonstrated that C/EBPβ-LncRNA187415.1-CISH maintained the pro-tumor characteristics of BCAMs and contributed to breast cancer progression. LncRNA187415.1 silence in BCAMs inhibited breast cancer cells proliferation, migration, and invasion, promoted apoptosis, and ameliorated breast cancer progression. Our results shed a light on immunotherapy of breast cancer targeting BCAMs. The authors have declared that no conflict of interest exists. This work was supported by Primary Research and Development Plan of Jiangsu Province (grant number: BE2019700), Jiangsu Province “333” project (grant number: BRA2018016), Six talent peaks project in Jiangsu Province (grant number: 2019-WSN-122), Projects of International Cooperation from Jiangsu (grant number: BX2019100), and International cooperation and exchange from Zhenjiang (grant number: GJ2020010). All animal procedures were in compliance with the Guide for the Care and Use of Laboratory Animals (NIH, 76 FR 91 [May 11, 2011]) and were approved by the Animal Care and Use committee of Jiangsu University. All authors approve to submit it as the present style. All the data were shown in the manuscript and are available from the correspondence author upon request. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.