Danhong Shen

Abstract Purpose: FGFR gene aberrations are associated with tumor growth and survival. We explored the role of FGFR2 amplification in gastric cancer and the therapeutic potential of AZD4547, a potent and selective ATP-competitive receptor tyrosine kinase inhibitor of fibroblast growth factor receptor (FGFR)1–3, in patients with FGFR2-amplified gastric cancer. Experimental Design: Array-comparative genomic hybridization and FISH were used to identify FGFR2 amplification in gastric cancer patient tumor samples. The effects of FGFR2 modulation were investigated in gastric cancer cells with FGFR2 amplification and in patient-derived gastric cancer xenograft (PDGCX) models using two approaches: inhibition with AZD4547 and short hairpin RNA (shRNA) knockdown of FGFR2. Results: Amplification of the FGFR2 gene was identified in a subset of Chinese and Caucasian patients with gastric cancer. Gastric cancer cell lines SNU-16 and KATOIII, carrying the amplified FGFR2 gene, were extremely sensitive to AZD4547 in vitro with GI50 values of 3 and 5 nmol/L, respectively. AZD4547 effectively inhibited phosphorylation of FGFR2 and its downstream signaling molecules and induced apoptosis in SNU-16 cells. Furthermore, inhibition of FGFR2 signaling by AZD4547 resulted in significant dose-dependent tumor growth inhibition in FGFR2-amplified xenograft (SNU-16) and PDGCX models (SGC083) but not in nonamplified models. shRNA knockdown of FGFR2 similarly inhibited tumor growth in vitro and in vivo. Finally, compared with monotherapy, we showed enhancement of in vivo antitumor efficacy using AZD4547 in combination with chemotherapeutic agents. Conclusion: FGFR2 pathway activation is required for driving growth and survival of gastric cancer carrying FGFR2 gene amplification both in vitro and in vivo. Our data support therapeutic intervention with FGFR inhibitors, such as AZD4547, in patients with gastric cancer carrying FGFR2 gene amplification. Clin Cancer Res; 19(9); 2572–83. ©2013 AACR.

In preclinical gastric cancer (GC) models, FGFR2 amplification was associated with increased tumour cell proliferation and survival, and drugs targeting this pathway are now in clinical trials.FGFR2 FISH was performed on 961 GCs from the United Kingdom, China and Korea, and the relationship with clinicopathological data and overlap with HER2 amplification were analysed.The prevalence of FGFR2 amplification was similar between the three cohorts (UK 7.4%, China 4.6% and Korea 4.2%), and intratumoral heterogeneity was observed in 24% of FGFR2 amplified cases. FGFR2 amplification was associated with lymph node metastases (P<0.0001). FGFR2 amplification and polysomy were associated with poor overall survival (OS) in the Korean (OS: 1.83 vs 6.17 years, P=0.0073) and UK (OS: 0.45 vs 1.9 years, P<0.0001) cohorts, and FGFR2 amplification was an independent marker of poor survival in the UK cohort (P=0.0002). Co-amplification of FGFR2 and HER2 was rare, and when high-level amplifications did co-occur these were detected in distinct areas of the tumour.A similar incidence of FGFR2 amplification was found in Asian and UK GCs and was associated with lymphatic invasion and poor prognosis. This study also shows that HER2 and FGFR2 amplifications are mostly exclusive.

Increased populations of regulatory T cells (Tregs) impair anti-tumor immunity. Recently, the transcription factor Foxp3 has been reported to play a key role in CD4(+)CD25(+) regulatory T cell function and represents a specific marker for these cells. However, Foxp3 is a nuclear protein and is of limited value in the isolation of Tregs, which is a major reason that many functionally relevant aspects of Treg cells are still unknown. Here, we have characterized CD4(+)CD25(+)CD127(low/)- as the surface marker of regulatory T cells in gastric cancer. 88.1-96.1%of CD25(+)CD127(low/-) T cells expressed Foxp3, the frequency of CD4(+)CD25(+)CD127(low/-) regulatory T cells in the peripheral blood of gastric cancer patients was significantly higher than that in healthy controls. Increased CD4(+)CD25(+)CD127(low/-) regulatory T cells were also present in the tumor microenvironment, such as those found in the ascites fluid, tumor tissue or adjacent lymph nodes. Particularly those Treg cells associated with the TNM stage. In addition, we found that CD4(+)CD25(+)CD127(low/-) Tregs suppressed effector T cell proliferation and also correlated to advanced stage of gastric cancer. Thus, CD4(+)CD25(+)CD127(low/-) can be used as a selective biomarker to enrich human Treg cells and also to perform functional in vitro assays in gastric cancer.

Gastric cancer (GC) is a leading cause of cancer deaths worldwide. Since the approval of trastuzumab, targeted therapies are emerging as promising treatment options for the disease. This study aimed to explore the molecular segmentation of several known therapeutics targets, human epidermal growth factor receptor 2 (HER2), MET and fibroblast growth factor receptor 2 (FGFR2), within GC using clinically approved or investigational kits and scoring criteria. Knowledge of how these markers are segmented in the same cohort of GC patients could improve future clinical trial designs.Using immunohistochemistry (IHC) and FISH methods, overexpression and amplification of HER2, FGFR2 and MET were profiled in a cohort of Chinese GC samples. The correlations between anti-tumour sensitivity and the molecular segments of HER2, MET and FGFR2 alterations were further tested in a panel of GC cell lines and the patient-derived GC xenograft (PDGCX) model using the targeted inhibitors.Of 172 GC patients, positivity for HER2, MET and FGFR2 alternations was found in 23 (13.4%), 21 (12.2%) and 9 (5.2%) patients, respectively. Positivity for MET was found in 3 of 23 HER2-positive GC patients. Co-positivity for FGFR2 and MET was found in 1 GC patient, and amplification of the two genes was found in different tumour cells. Our study in a panel of GC cell lines showed that in most cell lines, amplification or high expression of a particular molecular marker was mutually exclusive and in vitro sensitivity to the targeted agents lapatinib, PD173074 and crizotinib was only observed in cell lines with the corresponding high expression of the drugs' target protein. SGC031, an MET-positive PDGCX mouse model, responded to crizotinib but not to lapatinib or PD173074.Human epidermal growth factor receptor 2, MET and FGFR2 oncogenic driver alterations (gene amplification and overexpression) occur in three largely distinct molecular segments in GC. A significant proportion of HER2-negative patients may potentially benefit from MET- or FGFR2-targeted therapies.

The basidiomycetous yeast Rhodosporidium paludigenum Fell & Tallman isolated from the south of East China Sea was evaluated for its activity in reducing postharvest decay of cherry tomatoes caused by Alternaria alternata in vitro and in vivo tests. The results showed that washed cell suspension of R. paludigenum provided better control of A. alternata than any other treatment, while the autoclaved cell culture failed to provide protection against the pathogen. The concentration of antagonist had significant effect on biocontrol effectiveness in vivo: when the concentration of the washed yeast cell suspension was used at 1 x 10(9)cells/ml, the percentage rate of black rot of cherry tomato fruit was only 37%, which was remarkably lower than that treated with water (the control) after 5days of incubation at 25 degrees C. Furthermore, a great biocontrol efficacy of R. paludigenum was observed when it was applied prior to inoculation with A. alternata: the longer the incubation time of R. paludigenum, the lower disease incidence would be. However, there was little efficacy when R. paludigenum was applied after A. alternata inoculation. In addition, on the wounds of cherry tomato, it was observed that R. paludigenum grew rapidly increasing 50-fold during the first 12h at 25 degrees C. To the best of our knowledge, this is a first report concerning that the marine yeast R. paludigenum could be used as a biocontrol agent of postharvest fungal disease.

Long noncoding RNAs (lncRNAs) play important roles in the regulation of genes involved in cell proliferation. We have previously sought to more globally understand the differences of lncRNA expression between human fetal heart and adult heart to identify some functional lncRNAs which involve in the process of heart repair. We found that a highly conserved long noncoding RNA NR_045363 was mainly expressed in cardiomyocytes and rarely in non-cardiomyocytes. NR_045363 overexpression in 7-day-old mice heart could improve cardiac function and stimulate cardiomyocyte proliferation after myocardial infarction. Furthermore, NR_045363 knockdown inhibited proliferation of primary embryonic cardiomyocytes, while NR_045363 overexpression enhanced DNA synthesis and cytokinesis in neonatal cardiomyocytes in vitro. Mechanistic analysis revealed that NR_045363 promoted cardiomyocyte proliferation through interaction with miR-216a, which regulated the JAK2-STAT3 pathway. Our results showed that NR_045363 is a potent lncRNA modulator essential for cardiomyocyte proliferation.

Experimental study of coherent structures downstream of a spacer grid with different inclination angles (20°, 25°, 30°) of split-type mixing vanes in a 5 × 5 rod bundle was conducted by time-resolved particle image velocimetry (TR-PIV) under Reynolds number from 6.6 × 103 to 3.96 × 104. The fluorinated ethylene propylene (FEP) tubes and water are used as the matched index of refraction (MIR) materials. The maximum standard uncertainty of time-averaged velocity is less than 2 % of bulk flow velocity (Wb), and that of Reynolds stress is less than 2 % Wb2. The secondary flow structures downstream spacer grid with mixing vanes of different inclination angles change with several similar patterns, including vortices shedding from mixing vanes and fast dissipation near spacer grid, counter-current shear-flow development and shear-flow induced vortices formation, shear-flow induced vortices decaying and dissipation. The development trends of spanwise time-averaged velocity, Reynolds stress, integral spatial length scales and integral temporal length scales are determined by coherent structures. With Reynolds number increasing, smaller coherent structures are generated and the time-averaged velocity, Reynolds stress, spatial and temporal length scales decrease. Larger inclination angle generates larger coherent structures, and spatial and temporal length scales increase with inclination angle increasing. The experimental findings provide valuable information for understanding sub-channel mixing phenomena caused by mixing vaned spacer grids. The experimental data can be used for sub-channel mixing model developments and validations of turbulent models.

Summary Type III secretion systems are found in many G ram‐negative bacteria. They are activated by contact with eukaryotic cells and inject virulence proteins inside them. Host cell detection requires a protein complex located at the tip of the device's external injection needle. The Shigella tip complex ( TC ) is composed of IpaD , a hydrophilic protein, and IpaB, a hydrophobic protein, which later forms part of the injection pore in the host membrane. Here we used labelling and crosslinking methods to show that TCs from a Δ ipaB strain contain five IpaD subunits while the TCs from wild‐type can also contain one IpaB and four IpaD subunits. Electron microscopy followed by single particle and helical image analysis was used to reconstruct three‐dimensional images of TCs at ∼20 Å resolution. Docking of an IpaD crystal structure, constrained by the crosslinks observed, reveals that TC organisation is different from that of all previously proposed models. Our findings suggest new mechanisms for TC assembly and function. The TC is the only site within these secretion systems targeted by disease‐protecting antibodies. By suggesting how these act, our work will allow improvement of prophylactic and therapeutic strategies.

Increasing evidence indicates that microRNAs (miRNAs) play important roles in Kawasaki disease (KD). Our previous study demonstrated that hsa-miR-27b-3p (miR-27b) was up-regulated in KD serum. However, the specific role of miR-27b in KD remains unclear. We aimed to investigate that miR-27b could be a biomarker and therapeutic target for KD treatment. As well, the specific mechanism of miR-27b effecting endothelial cell functions was studied.The expression of miR-27b and Smad7 was measured by qRT-PCR. Gain-of-function strategy was used to observe the effect of miR-27b on human umbilical vein endothelial cells (HUVECs) proliferation and migration. Bioinformatics analyses were applied to predict miR-27b targets and then we verified Smad7 by a luciferase reporter assay. Western blot was performed to detect the protein expression of Smad7, PCNA, MMP9, MMP12 and TGF-β-related genes.We confirmed that miR-27b was shown to be dramatically up-regulated in KD serum and KD serum-treated HUVECs and that elevated expression of miR-27b suppressed the proliferation and migration of HUVECs. Furthermore, our results verified that miR-27b mediated cell functions by affecting the TGF-β via targeting Smad7 in HUVECs.These results suggested that up-regulated miR-27b had a protective role in HUVECs proliferation and migration via targeting Smad7 and affecting TGF-β pathway. Therefore, miR-27b represented a potential biomarker for KD and may serve as a promising therapeutic target for KD treatment.

Abstract Acute myocardial ischaemia/reperfusion (MI/R) injury causes severe arrhythmias with a high rate of lethality. Extensive research focus on endoplasmic reticulum (ER) stress and its dysfunction which leads to cardiac injury in MI/R Our study evaluated the effects of sulodexide (SDX) on MI/R by establishing MI/R mice models and in vitro oxidative stress models in H9C2 cells. We found that SDX decreases cardiac injury during ischaemia reperfusion and decreased myocardial apoptosis and infarct area, which was paralleled by increased superoxide dismutase and reduced malondialdehyde in mice plasm, increased Bcl‐2 expression, decreased BAX expression in a mouse model of MI/R. In vitro, SDX exerted a protective effect by the suppression of the ER stress which induced by tert‐butyl hydroperoxide (TBHP) treatment. Both of the in vivo and in vitro effects were involved in the phosphatidylinositol 3‐kinase (PI3K)/Akt signalling pathway. Inhibition of PI3K/Akt pathway by specific inhibitor, LY294002, partially reduced the protective effect of SDX. In short, our results suggested that the cardioprotective role of SDX was related to the suppression of ER stress in mice MI/R models and TBHP‐induced H9C2 cell injury which was through the PI3K/Akt signalling pathway.

ABSTRACT The type III secretion system (TTSS) of Pseudomonas aeruginosa is induced in vivo upon contact with eukaryotic cells and in vitro by calcium depletion in culture medium. We have observed a previously identified protein, PsrA, necessary for full activation of TTSS gene expression in P. aeruginosa . Electrophoretic mobility shift assays showed that recombinant PsrA could bind to the exsCEBA promoter region. A mutant with a deletion in the psrA gene was constructed. Using transcriptional fusions, we demonstrated that PsrA is required for the full activation of transcription of the TTSS regulatory operon exsCEBA and effector exoS , although the deletion mutant still responded to calcium depletion, to serum, and to host cell contact. The psrA mutant showed a marked decrease in the secretion of the type III effectors and weak resistance to phagocyte-like PLB-985 cells. The defect in TTSS transcription and secretion in the psrA mutant could be complemented by expression in trans of psrA . PsrA was previously identified as a transcriptional activator of RpoS, a central regulator during stationary phase. We confirmed with our strain that RpoS has a negative effect on TTSS gene expression. Taken altogether, these results suggest that PsrA is a newly identified activator that is involved in the expression of the TTSS by enhancing the exsCEBA transcriptional level.

ABSTRACT Type III secretion systems (T3SSs) are key determinants of virulence in many Gram-negative bacterial pathogens. Upon cell contact, they inject effector proteins directly into eukaryotic cells through a needle protruding from the bacterial surface. Host cell sensing occurs through a distal needle “tip complex,” but how this occurs is not understood. The tip complex of quiescent needles is composed of IpaD, which is topped by IpaB. Physical contact with host cells initiates secretion and leads to assembly of a pore, formed by IpaB and IpaC, in the host cell membrane, through which other virulence effector proteins may be translocated. IpaB is required for regulation of secretion and may be the host cell sensor. It binds needles via its extreme C-terminal coiled coil, thereby likely positioning a large domain containing its hydrophobic regions at the distal tips of needles. In this study, we used short deletion mutants within this domain to search for regions of IpaB involved in secretion regulation. This identified two regions, amino acids 227 to 236 and 297 to 306, the presence of which are required for maintenance of IpaB at the needle tip, secretion regulation, and normal pore formation but not invasion. We therefore propose that removal of either of these regions leads to an inability to block secretion prior to reception of the activation signal and/or a defect in host cell sensing.

Type III secretion systems (T3SSs) are essential devices in the virulence of many Gram-negative bacterial pathogens. They mediate injection of protein effectors of virulence from bacteria into eukaryotic host cells to manipulate them during infection. T3SSs involved in virulence (vT3SSs) are evolutionarily related to bacterial flagellar protein export apparatuses (fT3SSs), which are essential for flagellar assembly and cell motility. The structure of the external and transmembrane parts of both fT3SS and vT3SS is increasingly well-defined. However, the arrangement of their cytoplasmic and inner membrane export apparatuses is much less clear. Here we compare the architecture of the cytoplasmic regions of the vT3SSs of Shigella flexneri and the vT3SS and fT3SS of Salmonella enterica serovar Typhimurium at ~5 and ~4 nm resolution using electron cryotomography and subtomogram averaging. We show that the cytoplasmic regions of vT3SSs display conserved six-fold symmetric features including pods, linkers and an ATPase complex, while fT3SSs probably only display six-fold symmetry in their ATPase region. We also identify other morphological differences between vT3SSs and fT3SSs, such as relative disposition of their inner membrane-attached export platform, C-ring/pods and ATPase complex. Finally, using classification, we find that both types of apparatuses can loose elements of their cytoplasmic region, which may therefore be dynamic.

Kawasaki disease (KD) is an acute, self‑limited vasculitis that predominantly affects medium‑sized arteries, particularly the coronary arteries. Recent studies have indicated that microRNAs are involved in many diseases, including KD. However, the detailed mechanism remains unclear. The aim of the present study was to explore the role of miR‑186 in KD and potentially discover a new target for KD treatment. The results demonstrated that miR‑186 was upregulated in serum from patients with KD and KD serum could increase miR‑186 transcript levels in endothelial cells (HUVECs). Overexpression of miR‑186 mimic induced HUVEC apoptosis through mitogen‑activated protein kinase (MAPK) activation by targeting and inhibiting SMAD family member 6 (SMAD6). Furthermore, KD serum induced HUVEC apoptosis through miR‑186. In conclusion, the present results suggested that KD serum‑associated miR‑186 has an essential role in endothelial cell apoptosis by activating the MAPK pathway through targeting the SMAD6 gene.

Type III secretion systems (T3SSs) are central virulence devices for many Gram-negative bacterial pathogens of humans, animals & plants. Upon physical contact with eukaryotic host cells, they translocate virulence-mediating proteins, known as effectors, into them during infection. T3SSs are gated from the outside by host-cell contact and from the inside via two cytoplasmic negative regulators, MxiC and IpaD in Shigella flexneri, which together control the effector secretion hierarchy. Their absence leads to premature and increased secretion of effectors. Here, we investigated where and how these regulators act. We demonstrate that the T3SS inner membrane export apparatus protein MxiA plays a role in substrate selection. Indeed, using a genetic screen, we identified two amino acids located on the surface of MxiA's cytoplasmic region (MxiAC) which, when mutated, upregulate late effector expression and, in the case of MxiAI674V, also secretion. The cytoplasmic region of MxiA, but not MxiAN373D and MxiAI674V, interacts directly with the C-terminus of MxiC in a two-hybrid assay. Efficient T3S requires a cytoplasmic ATPase and the proton motive force (PMF), which is composed of the ΔΨ and the ΔpH. MxiA family proteins and their regulators are implicated in utilization of the PMF for protein export. However, our MxiA point mutants show similar PMF utilisation to wild-type, requiring primarily the ΔΨ. On the other hand, lack of MxiC or IpaD, renders the faster T3S seen increasingly dependent on the ΔpH. Therefore, MxiA, MxiC and IpaD act together to regulate substrate selection and secretion mode in the T3SS of Shigella flexneri.

As a green and renewable energy source, hydrogen can be produced by the electrolysis of water via the hydrogen evolution reaction (HER). Nevertheless, this method requires efficient and low-cost electro-catalysts to improve hydrogen production efficiency. Ionic liquids (ILs), with a unique combination of such superior properties as low vapor pressure, high electrical conductivity, high electrochemical stability, and a wide variety of functional groups, have found applications in electrochemical systems designed for efficient HER. Herein, we provide a comprehensive and updated review on the functions and performance of ILs used in electrochemical systems to enhance the HER. As the name suggests, ILs have been employed either as electrolytes by themselves, or as electrolyte additives. They also played many functional roles in the synthesis of HER electrocatalysts, including as the synthesis reaction solvent, reaction precursor as well as single/dual ion sources, binder and structure-directing agents of the catalysts. With the assistance of ILs, HER efficiency of electrocatalysts was improved significantly, resulting in decreased overpotentials in the range of 16-385 mV @ 10 mA cm-2 and increased Tafel slopes in the range of 30-210 mV dec-1. Lastly, the problems and challenges of ILs in electrocatalytic water electrolysis and HER are also discussed and their prospects considered.

The Pseudomonas aeruginosa type III secretion system (T3SS) is known to be a very important virulence factor in acute human infections, but it is less important in maintaining chronic infections in which T3SS genes are downregulated. In vitro, the activation of T3SS expression involves a positive activating loop that acts on the transcriptional regulator ExsA. We have observed that in vivo T3SS expression is cell density-dependent in a manner that does not need known quorum-sensing (QS) signals. In addition, stationary-phase culture supernatants added to exponential-phase growing strains can inhibit T3SS expression. The analysis of transposon insertion mutants showed that the production of such T3SS-inhibiting signals might depend on tryptophan synthase and hence tryptophan, which is the precursor of signalling molecules such as indole-3-acetic acid (IAA), kynurenine and Pseudomonas quinolone signal (PQS). Commercially available tryptophan-derived molecules were tested for their role in the regulation of T3SS expression. At millimolar concentrations, IAA, 1-naphthalacetic acid (NAA) and 3-hydroxykynurenine inhibited T3SS expression. Inactivation of the tryptophan dioxygenase-encoding kynA gene resulted in a decrease in the T3SS-inhibiting activity of supernatants. These observations suggest that tryptophan catabolites are involved in the downregulation of T3SS expression in the transition from a low- to a high-cell-density state.

Aim: To investigate the significance of epidermal growth factor receptor (EGFR), human epidermal growth factor receptor (HER)2, and HER3 expression on survival outcomes in Chinese gastric cancer patients. Materials and methods: Formalin-fixed, paraffin-embedded specimens from 121 patients who underwent gastrectomy at Shanghai Renji Hospital from 2007–2010 were retrospectively examined. Fluorescence in situ hybridization and immunohistochemistry techniques were used to identify gene amplification and protein overexpression. Correlations between the expression or amplification of HER family genes and clinicopathological parameters were then determined using statistical analysis. Results: EGFR protein overexpression, an increase in HER2 copy number and gene amplification, and HER3 protein overexpression were identified in 33.1%, 17.4%, and 62.0% of samples, respectively. Statistical analysis showed a significant association between EGFR expression and tumor invasion depth or tumor stage. HER2 was also shown to be significantly associated with the tumor grade. In addition, EGFR protein overexpression was found to be significantly associated with worse overall survival ( P =0.03). Conclusion: The HER family members showed a high expression in gastric cancer. EGFR protein expression was associated with overall survival. Keywords: gastric cancer, clinicopathologic significance, immunohistochemistry, fluorescence in situ hybridization

Type III secretion systems are complex nanomachines used for injection of proteins from Gram-negative bacteria into eukaryotic cells. Although they are assembled when the environmental conditions are appropriate, they only start secreting upon contact with a host cell. Secretion is hierarchical. First, the pore-forming translocators are released. Second, effector proteins are injected. Hierarchy between these protein classes is mediated by a conserved gatekeeper protein, MxiC, in Shigella As its molecular mechanism of action is still poorly understood, we used its structure to guide site-directed mutagenesis and to dissect its function. We identified mutants predominantly affecting all known features of MxiC regulation as follows: secretion of translocators, MxiC and/or effectors. Using molecular genetics, we then mapped at which point in the regulatory cascade the mutants were affected. Analysis of some of these mutants led us to a set of electron paramagnetic resonance experiments that provide evidence that MxiC interacts directly with IpaD. We suggest how this interaction regulates a switch in its conformation that is key to its functions.

The combination of antiapoptotic and angiogenic actions may represent a pharmacotherapeutic strategy for the treatment of myocardial infarction. Fibroblast growth factor (FGF) is expressed in various cell types including endothelial and muscle cells and promotes their survival, migration, and proliferation.Myocardial microvascular endothelial cells were divided into four treatment groups, the sham, hypoxia, basic FGF (bFGF), and bFGF plus 2-methoxyestradiol groups, and subjected to in vitro apoptotic analysis and Matrigel assays. An in vivo model of myocardial infarction was established by ligaturing the left coronary artery of mice in the four treatment groups. Cardiac performance, myocardial injury, endothelial cell angiogenesis, and myocardial apoptosis were assessed. bFGF administration after myocardial infarction improved cardiac function and cell viability, attenuated myocardial injury and apoptosis, and enhanced angiogenesis. Western blotting of HIF-1α, p-AKT, VEGF, p53, BAX, and Bcl-2 showed that bFGF increased HIF-1α, p-AKT, VEGF, and Bcl-2 and decreased BAX protein levels.The results of the present study indicated that bFGF attenuates myocardial injury by inhibiting apoptosis and promoting angiogenesis via a novel HIF-1α-mediated mechanism and a potential utility of bFGF in protecting against myocardial infarction.

Abstract Deciphering translation is of paramount importance for the understanding of many diseases, and antibiotics played a pivotal role in this endeavour. Blasticidin S (BlaS) targets translation by binding to the peptidyl transferase center of the large ribosomal subunit. Using biochemical, structural and cellular approaches, we show here that BlaS inhibits both translation elongation and termination in Mammalia. Bound to mammalian terminating ribosomes, BlaS distorts the 3′CCA tail of the P-site tRNA to a larger extent than previously reported for bacterial ribosomes, thus delaying both, peptide bond formation and peptidyl-tRNA hydrolysis. While BlaS does not inhibit stop codon recognition by the eukaryotic release factor 1 (eRF1), it interferes with eRF1’s accommodation into the peptidyl transferase center and subsequent peptide release. In human cells, BlaS inhibits nonsense-mediated mRNA decay and, at subinhibitory concentrations, modulates translation dynamics at premature termination codons leading to enhanced protein production.

Sub-channel code is one of the well-applied numerical tools in nuclear reactor thermal-hydraulics analysis. It takes consideration of the lateral transfer between adjacent sub-channels, which is its distinct characteristic. In most sub-channel codes, only one turbulent mixing coefficient for energy is used to account for lateral turbulent exchange. This imperfect description of the turbulent mixing parameter in different equations (e.g. mass, momentum and energy) significantly affects the accuracy of the calculation result. Besides, the empirical correlations to get the value of this coefficient have limited parameter ranges. In this paper, CFD simulations of two sub-channels in bare rod were performed with large geometry and flow condition ranges. The SSG turbulent model was used to simulate the non-isotropic turbulence and the calculation result was verified with experimental data. Based on the phenomenological analysis and theoretical consideration, a new turbulent mixing modeling approach was developed by studying βm, βE and βM, the turbulent mixing coefficients for mass, energy and momentum, respectively. Three empirical correlations of these coefficients were suggested and compared with existing correlations.

In this study, an electrocatalytic system using reduced graphene oxide supported single-atom copper (SA-Cu/rGO) anode was constructed for perfluorooctanoic acid (PFOA) defluorination. The successful doping of SA-Cu elevated the oxygen evolution potential and electrochemical active surface area of the anode and provided high electrocatalytic properties. Under optimum initial conditions with a peroxymonosulfate (PMS) concentration of 10 mM, a current density of 15 mA·cm−2, and an unadjusted pH of 7.75, the SA-Cu/rGO system achieved 98.8% removal ratio, 92.5% defluorination, and 94.4% mineralization of 20 mg·L-1 PFOA in 120 min. Kinetic studies indicate PFOA degradation following a pseudo-first-order model, with a rate constant of 4.7 × 10−2 min−1. The presence of PMS in the electrocatalytic system significantly improved the degradation rate and defluorination ratio. Radical trapping and quenching experiments confirmed that both ·OH and ·SO4- radicals contributed to PFOA degradation, with ·SO4- playing a more important role in defluorination. The system show high flexibility in initial PFOA concentrations (0.2–20 mg·L−1) and environmental pH values (3.0–10.0), as well as good durability. The electrocatalytic degradation mechanism of PFOA was proposed through determination of intermediates and density functional theory (DFT) calculation. The total short-chain perfluorocarboxylic acids (PFCAs) intermediates generated throughout the degradation were far from sufficient recovery of the PFOA loss, indicating that apart from the ·OH mediated decarboxylation-hydroxylation-elimination-hydrolysis (DHEH) pathway, the ·SO4- mediated decarboxylation-hydroxylation-oxidation-decarbonyl fluoride (DHOD) pathway may play a significant role in the near-complete defluorination and mineralization of PFOA. Moreover, DFT calculations suggest that the DHOD pathway is energetically favorable over DHEH. This study provides a further understanding of the defluorination mechanism of PFOA in electrocatalytic systems and demonstrates that electrochemical degradation with single-atom copper catalyst is an promising method for PFOA remediation in aqueous environment.

Purpose This study aimed to evaluate the impact of intravitreal triamcinolone acetonide (TA) administration after peeling of idiopathic epiretinal membranes (iERM) on both anatomical and visual outcomes, utilizing the ectopic inner foveal layer (EIFL) staging scheme. Methods In this retrospective case-control study, we analyzed 43 eyes from 43 patients diagnosed with iERM between June 2019 and December 2021. All participants were categorized into the TA or control groups based on administering intravitreal TA injection following ERM peeling. We thoroughly reviewed the clinical data, including the preoperative and postoperative best-corrected visual acuity (BCVA), central foveal thickness (CFT), and macular cube volume (VOL), with ERM stages classified according to the EIFL staging scheme. Results The study enrolled 22 eyes in the TA and 21 in the control groups. Following a mean follow-up period of 11.07 ± 2.02 months, noteworthy improvements in EIFL stages were observed in both cohorts ( p &lt; 0.01), but without significant distinctions between groups. In the TA group, 63.64% of eyes demonstrated improvements in EIFL stages, while the control group exhibited 76.19% ( p = 0.37). At the final visit, both groups experienced a noteworthy reduction in the postoperative CFT and VOL ( p &lt; 0.05), coupled with significant improvement in BCVA ( p &lt; 0.01). No substantial differences appeared between the two groups concerning BCVA, CFT, and VOL (all p &gt; 0.05). Conclusions Our study suggested that concurrent intravitreal TA injection following ERM removal did not provide additional benefits regarding anatomical and visual improvement in iERM cases classified as Stages 2 and 3.

Pseudomonas aeruginosa is a ubiquitous and opportunistic pathogen that uses the type III secretion system (TTSS) to inject effector proteins directly into the cytosol of target cells to subvert the host cell's functions. Specialized bacterial chaperones are required for effective secretion of some effectors. To identify the chaperone of ExoS, the representative effector secreted by the TTSS of P. aeruginosa, we analyzed the role of a postulated chaperone termed Orf1. By allelic exchange, we constructed the mutant with the deletion of gene Orf1. Analysis of secreted and cell-associated fractions was performed by SDS-PAGE and Western blotting. Using strain expressing in trans Orf1, tagged by V5 polypeptide and histidine, protein-protein interaction was determined by affinity resin pull-down assay in combination with MALDI-TOF. The role of Orf1 in the expression of exoS was evaluated by gene reporter analysis. Pull-down assay showed that Orf1 binds to ExoS and ExoT. Secretion profile analysis showed that Orf1 was necessary for the optimal secretion of ExoS and ExoT. However, Orf1 had no effect on the expression of exoS. Orf1 is important for the secretion of ExoS probably by maintaining ExoS in a secretion-competent conformation. We propose to name Orf1 as SpcS for “specific Pseudomonas chaperone for ExoS”.

The type III secretion apparatus (T3SA), which is evolutionarily and structurally related to the bacterial flagellar hook basal body, is a key virulence factor used by many gram-negative bacteria to inject effector proteins into host cells. A hollow extracellular needle forms the injection conduit of the T3SA. Its length is tightly controlled to match specific structures at the bacterial and host-cell surfaces but how this occurs remains incompletely understood. The needle is topped by a tip complex, which senses the host cell and inserts as a translocation pore in the host membrane when secretion is activated. The interaction of two conserved proteins, inner-membrane Spa40 and secreted Spa32, respectively, in Shigella, is proposed to regulate needle length and to flick a type III secretion substrate specificity switch from needle components/Spa32 to translocator/effector substrates. We found that, as in T3SAs from other species, substitution N257A within the conserved cytoplasmic NPTH region in Spa40 prevented its autocleavage and substrate specificity switching. Yet, the spa40(N257A) mutant made only slightly longer needles with a few needle tip complexes, although it could not form translocation pores. On the other hand, Δspa32, which makes extremely long needles and also formed only few tip complexes, could still form some translocation pores, indicating that it could switch substrate specificity to some extent. Therefore, loss of needle length control and defects in secretion specificity switching are not tightly coupled in either a Δspa32 mutant or a spa40(N257A) mutant.

Gastrointestinal stromal tumor (GIST), the most common type of mesenchymal tumors of the gastrointestinal tract, is a recently recognized tumor. The biological behavior of GIST is highly variable. Surgical resection remains the major treatment for GIST. In this study we retrospectively analyzed our surgical experience with 181 GIST patients to determine the effects of the treatment and the pathological features and prognosis factors of these GIST patients.The clinicopathological features and follow-up data of the 181 patients with GIST who had received surgical resection between January 1999 and December 2007 at Ren Ji Hospital were retrospectively reviewed. Immunohistochemical stains including CD117 (KIT), CD34, and other markers were used. Tumor size, mitotic index and other pathological parameters were recorded. According to the consensus of NIH risk-group stratification system based on maximum tumor size and mitotic index (per 50 high power field), tumors were classified into very-low-risk group (15 tumors, 8.3%), low-risk group (48, 26.5%), intermediate-risk group (52, 28.7%) and high-risk group (66, 36.5%). Prognostic factors were analyzed by Cox analysis including age, sex, tumor size, tumor site, mitotic index, NIH categories and surgical procedures.One hundred and seven (59.1%) of the 181 tumors were located in the stomach, 51 (28.2%) in the small intestine, 9 (5.0%) in the colon and rectum, and 14 (7.7%) in other sites including the omentum and mesentery. The median age of the patients was 58 (range, 24-84) years, and 102 patients (56.4%) were male. Tumor size ranged from 0.5 to 30 cm, while the mean size was 7.02 cm. Metastasis was found in 7 patients. One hundred and seventy-six (97.2%) of the 181 patients underwent radical resection, and among them 26 patients received extensive resection with the adjacent organ adherent to the tumors. The positive rate for the KIT protein (CD117) in immunostaining was 94.5% (171/181), while that for CD34 was 86.2% (156/181). The 1-, 3-, and 5-year survival rates of the 181 patients were estimated to be 95.2%, 87.9% and 78.5%, respectively. There was a significant difference in age, tumor size, tumor site, mitotic index, NIH categories, and presence or absence of multivisceral resection (P<0.05). But there was no significant difference in sex between the groups. Cox hazard proportional model revealed that advanced clinical stage and large tumor size contributed to worse prognosis. The patients who were treated with imatinib because of recurrence and metastasis or high recurrence risk showed stable disease.Surgical resection is the gold standard of treatment for primary GIST. NIH categorization is simple and effective to evaluate GIST behavior and prognosis. Targeted therapy such as imatinib, a KIT tyrosine kinase inhibitor, may play an important role in the treatment of GIST.

Recent studies showed that diffuse large B-cell lymphoma (DLBCL) could be classified into germinal centre B cell-like (GCB) and non-germinal centre B cell-like (non-GCB) phenotypes according to CD10,Bcl-6 and MUM1 expression. But primary gastrointestinal DLBCL has rarely been studied. This study was aimed to investigate the relationship between immunophenotypic classification, therapeutic outcomes and the prognosis of patients with primary gastrointestinal DLBCL.Between 1998 and 2010, there were 151 patients studied at Shanghai Renji Hospital with a histopathological diagnosis of primary gastrointestinal DLBCL. Immunohistochemistry was performed using EnVision methods for CD10, BCL-6 and MUM1. The clinicopathologic features and follow-up data were analyzed by the Kaplan-Meier method, log-rank test and χ2 test.According to the expression of CD10, BCL-6 and MUM1, 31.8 % (48/151) of the cases belonged to the GCB subtype and 68.2 % (103/151) belonged to the non-GCB subtype. There was a significant difference of local lymph node metastasis between the GCB and non-GCB groups (P < 0.05). Patients in the GCB group had a better survival rate than those in the non-GCB group (5-year survival rate, 65.2 % vs 36.4 %, P < 0.05). In the GCB group, there was no significant difference in survival rates in patients receiving R-CHOP and CHOP therapy (P > 0.05). In the non-GCB group, the survival rate in patients treated with R-CHOP therapy was significantly longer than those treated with CHOP therapy (5-year survival rate, 62.8 % vs 30.8 %, P < 0.05).The immunophenotype classification of gastrointestinal DLBCL, which is closely related to local lymph node metastasis, is found to have prognostic significance. Immunophenotype classification is also useful in selecting the chemotherapy protocol.

T3SSs are essential virulence determinants of many Gram-negative bacteria, used to inject bacterial effectors of virulence into eukaryotic host cells. Their major extracellular portion, a ∼50 nm hollow, needle-like structure, is essential to host cell sensing and the conduit for effector secretion. It is formed of a small, conserved subunit arranged as a helical polymer. The structure of the subunit has been studied by electron cryomicroscopy within native polymers and by solid-state NMR in recombinant polymers, yielding two incompatible atomic models. To resolve this controversy, we re-examined the native polymer used for electron cryomicroscopy via surface labelling and solid-state NMR. Our data show the orientation and overall fold of the subunit within this polymer is as established by solid-state NMR for recombinant polymers.

Abstract Ataxia telangiectasia mutated (ATM) is a protein kinase that regulates cell-cycle checkpoints, DNA repair and recombination (1). It has been shown that ATM interacts with and regulates NBS1 and BRCA complex which is essential for the homologous recombination (HR) repair in response to the DNA double-strand break (DSB) damage (2). These observations suggest that tumor cells with defective ATM expression or activity subsequently leading to HR deficiency are likely more sensitive to the targeted therapeutic/chemotherapeutic agents that cause the accumulation of DNA DSBs during replication. To test this, we studied the growth inhibitory effects of olaparib (AZD2281; KU-0059436) and SN-38 (the active metabolite of Irinotecan) on gastric cancer (GC) cells with reduced ATM expression. Olaparib is a potent oral inhibitor of poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP), which has selective antitumor activity in cancers associated with BRCA1 and BRCA2 mutations (3). Our study here showed that 6 out of 7 GC cell lines with low or no expression of ATM are sensitive to olaparib inhibition with IC50s ≤ 1uM as measured by clonogenic survival assays. In contrast, the majority of other GC cell lines positive for ATM expression (11 out of 13) are less sensitive (1uM &amp;lt; IC50 ≤ 1.5uM) or resistant (IC50 &amp;gt; 1.5uM) to olaparib with only 2 of 13 lines sensitive. Similar correlation was also observed in these GC cell lines between reduced ATM expression and sensitivity to SN-38 treatment. In addition, combination of olaparib and SN-38 had a synergistic and selective inhibitory effect on cell growth of GC cells with low/no expression of ATM. These data suggest that loss of ATM in GC cell can sensitize their cellular response to olaparib or/and SN-38. Next, we asked whether this finding has clinical relevance in GC therapy. The tumor and tumor adjacent tissue specimens from more than 100 patients with gastric cancer were analyzed for ATM expression by immunohistochemistry. 22% (24/111) of the gastric tumor specimens were found stained ATM negative while the negative rate in the adjacent tissues was only 4% (1/26). These results suggested that loss of ATM expression is frequently associated with gastric tumorigenesis. Loss of ATM function may serve as an important tumor biomarker in GC and a PARP inhibitor as monotherapy or in combination with chemotherapy provides a promising therapeutic in this disease segment. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):B42.

Testicular torsion is a urology urgent disease which causes testicular injury and potential sterility. In this study, we explored the protective influence of rosiglitazone on testicular ischaemia–reperfusion damage. There were 28 male Sprague Dawley rats in total, which were assigned randomly to four groups. Group A was blank control one; group B was testicular injury one; group C was rosiglitazone one; group D was rosiglitazone antagonist one. The testicles were counter-rotated after 2 hr and then underwent orchiectomy 24 hr later. We found that testicular tissue structure of rats was seriously damaged in groups B and D. However, group C had better testicular architecture. Similar findings were also shown for lipid peroxidation by evaluating the MDA activity (p < .05). Unlike group B or group D, the levels of inflammation by evaluating the MPO activity, the levels of TNF-a, IL-1 and IL-6 and the expressions of ICAM-1 were prominently lower in group C (p < .05) as well. So our researches demonstrated that rosiglitazone significantly decreased the amount of responsive oxygen radical and regulated inflammatory responses. Rosiglitazone had a protective influence against testicular ischaemia–reperfusion injury in rats and possibly depended on its anti-inflammatory and antioxidant traits.

CO2 has anesthetic potency and effectively influences the circulatory system. We investigated the effects of Etco2 on the minimum alveolar concentration of sevoflurane that blunts the adrenergic response to surgical incision (MAC-BAR) in patients undergoing radical surgery for gastric carcinoma.Ninety patients undergoing radical gastric-carcinoma surgery under general anesthesia were enrolled and randomly assigned into 3 groups. After intubation, the Etco2 in group L (n = 30), group N (n = 30), and group H (n = 30) was adjusted to 25 mm Hg ≤ Etco2 <30 mm Hg, 30 mm Hg ≤ Etco2 < 40 mm Hg, and 40 mm Hg ≤ Etco2 < 45 mm Hg, respectively, by changes in controlled ventilation. Hemodynamics and depth of anesthesia were observed before and after skin incision. The MAC-BAR of sevoflurane for each group was determined using an up-and-down sequential-allocation technique.To obtain 7 crossovers, 25, 26, and 26 patients were used in group L, group N, and group H, respectively. The MAC-BAR of sevoflurane using the up-and-down method for group H was significantly lower than that for group L (2.3% [95% confidence interval {CI}, 2.2-2.4] vs 2.9% [95% CI, 2.7-3.0]; difference, -0.6% [95% CI, -0.7 to -0.4], P < .001) and group N (2.3% [95% CI, 2.2-2.4] vs 2.8% [95% CI, 2.8-2.9]; difference, -0.5% [95% CI, -0.7 to -0.4], P < .001), while no significant difference was found between group L and group N (P = 1.000).Higher Etco2 levels (Etco2 values equal to 40 mm Hg or higher) can effectively decrease the MAC-BAR of sevoflurane in patients undergoing radical surgery for gastric carcinoma.

Background: Stimulator of interferon gene (STING)-associated vasculopathy with onset in infancy (SAVI), caused by gain-of-function mutations in human transmembrane protein 173 ( TMEM173 ), is characterized by widespread chronic inflammation primarily affecting the skin and lungs. Although SAVI is an inflammatory disease, typical anti-inflammatory agents have limited or no effect. Methods and Results: A 1-year-old boy presented with recurrent facial rashes since he was 8 months. Moreover, he suffered from recurrent oral ulcers, chronic cough, and failure to thrive. Laboratory parameters showed elevated erythrocyte sedimentation rate (ESR) and immunoglobulin levels. Chest high-resolution computed tomography (HRCT) showed interstitial lung disease (ILD). Whole-exome sequencing revealed a heterozygous mutation in the TMEM173 gene (c.463G &gt; A, p.V155M). Ultimately, the patient was diagnosed with SAVI. Tofacitinib was initiated at the age of 19 months, resulting in the alleviation of facial rashes and improvement of ILD within 3 months. Conclusion: SAVI is a difficult-to-treat type I interferonopathy. We hope that JAKi treatment will prove valuable for SAVI patients.

Abstract The interplay of small, noncoding microRNAs (miRNAs), mRNAs and proteins plays crucial roles in almost all cellular processes. MiR-124, widely known as a memory-related miRNA, can regulate LTM by binding to the mRNA of CREB1 stimulated with 5-HT. In this paper, we establish a regulatory network model of CREB1 and miR-124 stimulated by 5-HT, in which miR-124 inhibits CREB1, which in turn enhances miR-124. Our model validates three protocols based on 5-HT in experiments on the induction of LTM in Aplysia . A steady-state analysis and numerical bifurcations of the abstracted system beyond memory formation, when the fast reaction has been in the equilibrium, can facilitate more abundant dynamical behaviors such as bistability and oscillation. The original system also exhibits bistability under appropriate feedback strengths, which is relevant to the mechanism of LTM formation. Furthermore, we specifically show a change in the transition from a reversible switch to an irreversible switch via bifurcations of the negative regulation of miR-124 on CREB1, which eventually maintains a high phosphorylated CREB1 level after initially elevated by 5-HT. These findings indicate that miR-124 provides an inhibitory constraint on long-term synaptic plasticity through the regulation of CREB1.

To study the effect of early intervention with lipoxin A4 (LXA4) on septic mice.Healthy male Balb/c mice aged 6-8 weeks were randomly divided into sham-operation group, sepsis group, 1-hour intervention group (intervention at 1 hour after sepsis), and 6-hour intervention group (intervention at 6 hours after sepsis) (n=8 each). A sepsis model was prepared by cecal ligation and puncture. The intervention groups received LXA4 at 0.01 μg/g body weight 1 or 6 hours after the model was established. Blood was taken from eyeballs at 24 hours after operation. Peritoneal lavage fluid and liver and lung tissue samples were collected. The bacterial colonies of whole blood and peritoneal lavage fluid were counted by dilution plating. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) were determined by cytometric bead array. The serum level of high mobility group box-1 (HGMB1) was determined using ELISA. The percentages of macrophages and neutrophils in peritoneal lavage fluid were determined by flow cytometry. Paraffin sectioning and hematoxylin-eosin staining were performed for the liver and lung tissue samples to observe pathological damage.Compared with the sham-operation group, the sepsis group had a significantly decreased percentage of macrophages and a significantly increased percentage of neutrophils in peritoneal lavage fluid (P<0.05), as well as significantly increased serum levels of IL-6, TNF-α, MCP-1, and HMGB1 (P<0.05); in addition, the sepsis group showed more vacuolar degeneration, hepatocyte swelling, and inflammatory cell infiltration in liver tissue, and more capillary congestion, pulmonary septal thickening, inflammatory cell infiltration, and partial tissue destruction in lung tissue. Compared with the sepsis group, the 1-hour and 6-hour intervention groups had a significantly increased percentage of macrophages in peritoneal lavage fluid (P<0.05) and significantly reduced bacterial load in whole blood (P<0.05), serum levels of IL-6, TNF-α, MCP-1, and HMGB1 (P<0.05), and degree of liver and lung tissue damage and inflammatory cell infiltration, but there was no significant difference in the percentage of neutrophils and bacterial load in peritoneal lavage fluid (P>0.05). Compared with the 6-hour intervention group, the 1-hour intervention group had a significantly decreased serum level of HMGB1 (P<0.05), but there was no significant difference in other indicators between the two groups (P>0.05).Early intervention with LXA4 may attenuate liver and lung injuries in septic mice, which may be explained by the decrease in serum levels of IL-6, TNF-α, MCP-1, and HMGB1, and it also may reduce the bacterial dissemination in the whole blood of septic mice, which may be explained by the increase in the percentage of peritoneal macrophages.

Twenty-five phenolic compounds in tobacco were analyzed using high performance liquid chromatography-ultraviolet/mass spectrometry (HPLC-UV/MS), including absolute quantification of 11 main phenols by HPLC-UV and relative quantification of other 14 phenols by SHPLC-MS. The validation results of this method were satisfactory. Eleven phenol standards showed good linearity with correlation coefficients (r2) of 0.999 3 - 0.999 9 over the mass concentration range from 0.90 mg/L to 99.00 mg/L (the range of chlorogenic acid and rutin was from 0.95 mg/L to 380.00 mg/L). The recoveries of the 11 phenol standards were 91.0% - 112.4% with the RSDs of 0.33% - 8.11% at the three spiked levels of 22.5 - 24.8, 45.0 - 49.5 and 67.5 - 74.3 mg/L. The reproducibility of the method was good with the RSDs of 1.48% - 13.40%. In addition, the intra-day and inter-day precisions were also satisfactory with the RSDs of 0.35% - 15.54%. Mature fresh tobacco leaves from Yunnan, Henan and Guizhou Provinces, China were analyzed using this method. The results showed that the total amount of main phenols was Guizhou > Yunnan > Henan and there was significant difference between Guizhou and Henan. The amount of rutin was Yunnan > Guizhou > Henan, and there were significant differences among them each other. The method is reproducible with a simple pretreatment covering a wide range of phenols, and can be applied in batch analysis of tobacco leaves.

The influence of the initial condition parameters on a free turbulent swirl diffusion flame and on its equivalent isothermal flow was experimentally studied. A parametric study was conducted on a 40 KW coaxial jet burner with a straight exit. The fuel used was methane. Both aerodynamic and flame structures were systematically and independently varied and data were collected for each characteristic condition. The experimental results were compared with existing data, obtained on burners with a diverging quarl. This showed that, besides the burner exit geometry, both fuel and air loading has a significant effect on the characteristics of the flow and its thermal structure. For the condition studied, an increase in the fuel loading or a decrease in the air loading was detrimental to the formation of a central recirculation zone, even leading to its disappearance. High turbulence levels were associated with the region close to the burner exit for high air and fuel loadings, and with the region where the central recirculation zone was steadily established. The flame temperature was reduced by high swirl, in opposition to what is commonly observed, since high swirl is, in general, associated with strong mixing and efficient combustion, and thus high flame temperature. Incomparison with the isothermal case, both favourable and unfavourable effects induced by the combustion were found on the formation of the central recirculation zone.

Recent studies have shown that diffuse large B-cell lymphoma (DLBCL) can be classified into germinal center B-cell-like (GCB) and non-GCB phenotypes by immunohistochemical staining. The aim of this study was to investigate the correlation of immunophenotypic classification with clinicopathological features in Chinese patients with primary gastric DLBCL to further our knowledge of this disease. Seventy-three patients with a histopathological diagnosis of primary gastric DLBCL were studied. Immunohistochemistry was carried out using the EnVision method to detect the expression of CD10, Bcl-6, and MUM1. The clinicopathologic features and follow-up data were analyzed using the Kaplan-Meier method, log-rank test, and χ (2) test. Expression of CD10 was observed in 21.9 % (16/73) of patients, Bcl-6 in 72.6 % (53/73), and MUM1 in 74.0 % (54/73). According to these data, 32.9 % (24/73) of the cases belonged to GCB subtype and 67.1 % (49/73) belonged to non-GCB subtype. There was a significant difference in tumor size and local lymph node metastasis between the GCB and non-GCB groups (P < 0.05). Complications in the GCB group (4.2 %) occurred less frequently than those in the non-GCB group (18.4 %); however, this difference was not significant (P > 0.05). Survival analysis revealed that patients in the GCB group had an increased 5-year survival rate compared to those in the non-GCB group (58.5 % vs 35.7 %, χ (2) = 3.939, P < 0.05). The 5-year survival rate of patients undergoing R-CHOP chemotherapy was significantly longer than that of patients in the CHOP group (74.7 % vs 37.5 %, χ (2) = 4.185, P < 0.05). The immunophenotype classification of primary gastric DLBCL, which is closely related to the tumor size and local lymph nodes metastasis, was found to have prognostic significance.

Objective To investigate the therapeutic experience of gastrointestinal stromal tumors (GIST) and to analyze the pathological features and prognostic factors of GIST. Methods The clinicopathologieal and follow-up data of 181 patients with GIST admitted in Renji Hospital between January 1999 and December 2007 were analyzed retrospectively. All the cases were grouped according to Fletcher's risk scheme. Life table and COX regression model were used to evaluate the prognostic factors. Results Out of 181 tumors, 107(59.1%) were located in stomach, 51 (28.2%) in intestine and 23 (12.7%) in colorectum or other sites. Distant metastases,including liver metastases were found in 7 patients intraoperatively. Tumor size ranged from 0.5 to 30 cm with the mean of 7.02 cm. The positive rate of CD117 was 94.5% (171/181) and that of CD34 was 86.2% (156/181). One hundred and seventy-six patients underwent complete resections, including multi-organ resections in 26 patients. The other patients underwent palliative operations. The 1-, 3-and 5-year overall survival rates of 181 patients were 95.2%, 87.9% and 78.5% respectively. Univariate analysis revealed age, tumor size, primary organ of tumor, mitotic count, Fletcher's classification and multi-organ resection were associated with survival rate. No significant difference of sex was existed among groups. COX hazard proportional model revealed that advanced stage and large tumor size indicated worse prognosis. Eight patients with high risk of recurrence and 3 patients with recurrence and metastasis were stable after receiving imatinib therapy. Conclusions The diagnosis of GIST depends on endoscope and CT. Fletcher's classification is simple and effective to evaluate GIST behavior and prognosis. Surgical resection is still the main therapy for GIST and targeted therapy will play a more important role for prognosis in the future. Key words: Gastrointestinal stromal tumors;  Surgical procedures;  Clinicopathology;  Targeted therapy;  Prognosis

The sub-channel analysis code is widely used in order to predict the thermal hydraulic performance in the core. Turbulent mixing presenting the influence of fluctuating velocity is one of the mechanisms of the lateral flow between sub-channels. The previous research on the turbulent mixing was focusing on the center to center sub-channels. In this study, based on the previous work, the turbulent mixing between different kinds of sub-channels is studied by CFD simulations. The results show that the coefficient β do not change much, if different kinds of sub-channels are considered. The coefficient βE and βM change in different ways because the distribution of temperature and velocity change in different ways. It is found that the influence of the hydraulic diameter is only related to Re number. The geometric shape factors are defined and their values are recommended according to the calculation results. Finally, when applied in the sub-channel code, the coefficients calculated by CFD method and the geometric shape factors have better performance than the previous turbulent mixing coefficient.

Background: Judges may experience mental health problems, which reduces their performance in juridical cases, and constitutes a great threat to both the authority of law and social justice. This study aimed to assess the prevalence of psychological distress among judges and examine the effect of perfectionism on psychological distress with age and gender as the moderators.&#x0D; Methods: This survey was carried out in person with 565 Chinese judges in 2016, using the Almost Perfect Scale-Revised and the Kessler Psychological Distress Scale. The effects of perfectionism on psychological distress was explored by Structural Equation Model (SEM), and the moderating effects of age and gender were tested by Multi-group analysis.&#x0D; Results: Psychological distress was reported by 89.20% of the judges surveyed. Discrepancy had a significant positive effect on psychological distress, but order had a significant negative effect on psychological distress, with high standards exerting no significant effect. Age had a significant moderating effect, whereas the moderating effect of gender was not significant.&#x0D; Conclusion: The three dimensions of perfectionism exerted different effects on psychological distress of Chinese judges, and the relationships were moderated by age.

Gastric neuroendocrine carcinoma (G-NEC) is a rare neoplasm known for its aggressive behavior and poor prognosis. Brunner's gland adenoma (BGA) is a rare benign proliferative lesion that develops most commonly in the duodenum. To the best of our knowledge, no cases of G‑NEC coexisting with BGA have previously been reported. The present study therefore reports the first case of G‑NEC combined with BGA. A 67‑year‑old female presented with upper abdominal discomfort. No distant metastases were detected upon pre‑operative abdominal computed tomography imaging. The patient underwent a radical distal gastrectomy, D2 lymphadenectomy and Billroth I gastroenterostomy. The resected masses were histologically confirmed to be G‑NEC and BGA, respectively. The patient did not receive neoadjuvant or adjuvant chemotherapy or radiotherapy, and remains alive with no evidence of metastasis or recurrence at four years post-surgery.

A kind of constant temperature and current operating high power LED module supplied by photovoltaic (PV)/wind/grid hybrid power system was designed in this paper. The LED was monitored while was controlled at room or lower temperature by a closed loop feedback circuit. An optimal PV/wind/grid complementary generation with lead-acid rechargeable battery was employed to supply the LED module. The maximum power point tracking (MPPT) scheme was adopted in the charging/discharging/unloading controller for the accumulator. Necessary computer software was programmed to perform technical tasks. The experimental result indicates that it's benefit to improving luminescence when the LED is controlled by the designed circuit at low temperature.

Objective. The aim of the study is to explore the relationship between the degree of illness in elderly patients with rheumatoid arthritis (RA) and parameters of musculoskeletal ultrasound (MSUS) and Oswestry Dysfunction Index (ODI) and its clinical value. Methods. The clinical data of 100 elderly patients with RA admitted to our hospital from May 2016 to May 2022 were retrospectively analyzed. The patients were divided into four groups, including the remission group (DAS28 ≤ 2.6, n = 25), low activity group (2.6 ≤ DAS28 ≤ 3.2, n = 25), middle activity group (3.2 ≤ DAS28 ≤ 5.1, n = 25), and high activity group (DAS &gt; 5.1, n = 25) according to the disease activity score-28 (DAS28). All patients underwent ultrasonic detection to compare the relationship between the degree of illness in elderly patients with RA and parameters of MSUS and ODI. Results. The total semiquantitative score of MSUS and ODI score in the remission group were obviously lower than those in the other three groups ( <math xmlns="http://www.w3.org/1998/Math/MathML" id="M1"> <mi>P</mi> <mo>&lt;</mo> <mn>0.001</mn> </math> ). The degree of illness in elderly patients with RA was positively correlated with parameters of MSUS (r = 0.886, <math xmlns="http://www.w3.org/1998/Math/MathML" id="M2"> <mi>P</mi> <mo>&lt;</mo> <mn>0.001</mn> </math> ). The degree of illness in elderly patients with RA was positively correlated with ODI (r = 0.907, <math xmlns="http://www.w3.org/1998/Math/MathML" id="M3"> <mi>P</mi> <mo>&lt;</mo> <mn>0.001</mn> </math> ). Conclusion. The degree of illness in elderly patients with RA is closely related to parameters of MSUS and ODI, and the parameters of MSUS have a higher evaluation value for the degree of illness in elderly patients with RA, which are correlated with ODI.

Objective To explore the protective role of lipoxin A4 (LXA4) during early process of atherosclerosis formation in rats with juvenile metabolic syndrome (MS). Methods Rat models of juvenile MS were established with 3-week Sprague-Dawley (SD) rats fed on high-carbonhydrates and high-fat diet for 6 weeks.The other quali-fied ones were randomly grouped into model group, LXA4 low-dose group, LXA4 middle-dose group, and LXA4 high-dose group, and a control group fed with normal forage.The low, middle, high-dose groups were injected different doses of LXA4 daily, while the model group and control group were injected with the same dose of isotonic NaCl solution for 2 consecutive weeks.After 2-week medication, the visceral adipose tissue were isolated by laparotomy and heart blood collected by thoracotomy under anesthesia, followed the fixation of thoracic and abdominal aortas in the immobilized rats.The mRNA expression level of inflammation cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP) in the adipose tissue were determined by semi-quantitative reverse transcription PCR (RT-PCR), respectively.Secretions of IL-6, and TNF-α in serum were determined by enzyme linked immunosorbent assay (ELISA). Immunocytochemistry was used to label endomembrane and middle-membrane of thoracic aorta, and endothelial cell layer in each group and the ratios of thickness of endomembrane and middle-membrane were compared. Results Compared with the control group, weight, body length and abdominal circumference of juvenile MS rats increased significantly (all P 0.05). Compared with control group, inflammatory cytokines IL-6, TNF-α secreted in serum of model rats were increased significantly (all P 0.05). Conclusions The increasing inflammatory cytokines are involved in early process of atherosclerosis formation in rats with juvenile MS.LXA4 by reducing the expression of inflammatory factor level in adipose tissue, thereby reducing the inflammatory cytokines in serum, alleviate the damage of arterial wall. Key words: Metabolic syndrome; Lipoxin A4; Inflammatory cytokine; Interleukin-6; Tumor necrosis factor-α; C-reactive protein; Atherosclerosis

Objective To observe the effects of high-fat feeding on growth and the expression of insulin-like growth factor 1(IGF-1), insulin receptor substrate 1(IRS-1) in adolescent rats' liver with non-alcoholic fatty liver disease, and to elucidate the relationship between growth failure in adolescent rats with non-alcoholic fatty liver disease and IGF-1, IRS-1 turbulence. Methods Thirty-six Sprague-Dawley (SD) young rats of 21 days were randomly divided into normal control group(NC group, n=18) and high-fat feeding group(HF group, n=18). Non-alcoholic fatty liver disease(NAFLD) model was induced by feeding the SD rats with high-fat food.Immunohistochemistry was applied to detect the expression levels of IGF-1 and IRS-1 in liver tissue.The expressions of mRNA of IGF-1 and IRS-1 were measured by RT-PCR. Results Compared with NC group, the serum alanine amino transferase(ALT), total cholesterol(TC), triglyceride(TG) of HF group in the 6th, 8th, 12th week were gradually increased.The serum ALT[(194.67±11.15)U/L], TC[(1.81±0.09)mmol/L], TG[(0.34±0.05)mmol/L]contents of HF group at 8th week were higher than those at 6th week[(166.00±22.01)U/L, (1.52±0.22)mmol/L, (0.41±0.12)mmol/L, respectively], and the serum ALT[(213.0±27.67)U/L], TC[(2.15±0.37)mmol/L], TG[(0.38±0.15)mmol/L]contents of the 12th week were significantly increased compared with 6th week and 8th week.The constitution and body length of the HF group were lower than those of the normal control group.With time extended, the liver tissue steatosis, inflammation, the balloon like change of the liver tissue pathology of HF group in 6, 8, 12th week gradually increased.Immunohistochemistry results showed that HF group IRS-1[(1.46±0.23), (0.74±0.17), (0.85±0.31)], IGF-1[(0.92±0.02), (0.83±0.02), (0.77±0.03)]expression gradually decreased, the difference was statistically significant(F=36.024, P<0.05). IGF-1 and IRS-1 mRNA expressions in HF group were consistent. Conclusion The liver tissue IGF-1 and IRS-1 are correlated with the weight and body length.The growth failure of young rats induced by high-fat feeding may be related to the decreased expression of IGF-1 and IRS-1. Key words: Fatty liver; Dietary fats, unsaturated; Insulin-like growth factor 1; Insulin receptor substrate proteins

To sub-clone and express the gene encoding Schistosoma japonicum calcium-binding protein (SjE16) and study its immunological response.The specific primers were designed according to the expressed sequence tags (ESTs) sequence, which was used for amplification of the encoding sequence from the cDNA clone containing SjE16. The gene was subcloned into pGEX4T-1 plasmid and expressed. The rSjE16 was tested for its immunological response by ELISA.The gene encoding Schistosoma japonicum SjE16 was cloned and expressed successfully. The immunogenicity and diagnostic potential of rSjE16 were investigated. It was demonstrated by immunoassay in rabbits that the specificity and sensitivity of the test were 94.1% (16/17) and 88.2% (15/17), respectively, and the level of antibody titer of the untreated group reached a peak at 9-11 wk post infection and maintained high at least for 21 wk post infection, while the antibody level in the treated group rapidly decreased to pre-infection level in 11 wk after treatment. In human, the specificity of the test was 98.3% (57/58); the sensitivity of acute and chronic patient serum assay was 85.5% (53/62) and 70.2% (40/57), respectively.The recombinant protein of SjE16 (rSjE16) was acquired. It can be recognized by the sera from schistosomiasis patients, and the level of antibodies decreased quickly after treatment in experimental rabbits, which implicates the potential value for the evaluation of chemotherapy and detection of active infection.

Objective To analyze the clinico-pathological characteristics and prognostic factors of patients with gastric neuroendocrine carcinoma(G-NEC).Methods Clinical data of 40 cases of G-NEC form January 2003 to August 2013 at Ren Ji Hospital of Shanghai Jiaotong University were analyzed.Tumors were classified into different grades and stages according to the 2010 WHO classification and the 2006 European neuroendocrine tumor society (ENETS).Follow-up was conducted by telephone.The survival curves were drawn using Kaplan-Meier method.Univariate analysis was performed by the Log-rank test and multivariate analysis was performed by the COX proportional hazards model.Results Among the 40 G-NECs patients,29 were male(72％) and 11 were female(28％),with an median age of 61 years.Tumors located in the gastric cardia in 20 cases,in the gastric antrum in 11 cases and in the gastric body in 9 cases.Tumor ranged from 1 cm-20 cm.All patients were G-NEC (G3).Follow-up rate was 100％ (40/40).The median overall survival rate was 12 months,and one-year survival rate was 82％.Immunohistochemically G-NEC cells were positive for CgA and Syn in 11 cases.Gender (x2 =5.673,P ＜ 0.05),Ki-67 index (x2 =8.612,P ＜ 0.05),and lymphnode involvement (x2 =0.559,P ＜ 0.05) were prognostic factors of G-NEC patients.Conclusions The symptoms of G-NEC are nonspecific.Its diagnosis relies on pathological examination and immunohistochemistry.Syn and CgA are the most important markers.Female gender,lower Ki-67 index and lower lymph node metastasis predict a survival advantage. Key words: Neuroendocrine tumors ;  Pathology, clinical;  Prognosis

Objective To compare the clinical efficacy of Freka trelumina (FT) vs. feeding jejunostomy (FJ) in carrying out postoperative early enteral nutrition (EEN) in old patients with gastric cancer. Method 168 old gastric cancer cases were devided into FT group (n=54) with EEN, FJ group (n=50) with gastric tube and EEN, and total parenteral nutrition (TPN) group (n=64). Results Compared with TPN group, postoperative body weight, serum albumin and prealbumin level in FT and FJ groups were significantly higher, intestinal function recovery time, days of postoperative hospitalization and costs were significantly lower. The incidence of cough, sputum and sore throat of patients in FT group were significantly higher than those in FJ and TPN groups (P<0.05). Conclusions Postoperative EEN through FT and FJ was effective to improve nutritional parameter, accelerate intestinal function recovery, reduce the number of days of postoperative hospitalization, total costs, anastomotic stomal leak and gastroparesis rate. Key words: Stomach neoplasms; Gastrectomy; Enteral nutrition

Objective To explore the clinicopathological and immunohistochemical features, diagnosis and treatment, prognosis of hepatoid adenocarcinoma of the stomach with neuroendocrine tumor. AFP, Syn, CgA, Ki-67, P53 were stained by immunohistochemistry and follow-up was conducted by telephone. Methods Hepatoid adenocarcinoma of the stomach with coexisting neuroendocrine tumor was identified in 13 cases from June 2004 to June 2014 in Ren Ji Hospital. Results Among the 13 cases of hepatoid adenocarcinoma of the stomach coexisting with neuroendocrine tumor patients, there were 7 males and 6 females, with an median age of 62 years. Tumors located in the gastric cardia in 2 cases, in the gastric antrum in 8 cases and in the gastric body in 3 cases.Tumor ranged from 1-19 cm. The follow-up rate was 100%. The median overall survival rate was 12 months, two patients died of liver metastasis and one patient died of anastomotic recurrence. Serum AFP increased in 10 patients with hepatoid adenocarcinoma of the stomach coexisted with neuroendocrine tumor. The structure consisted of hepatoid cell differentiation and adenocarcinoma differentiation and neuroendocrine differentiation area by histological microscope examination. Immunohistochemical staining showed that tumor regional AFP, Syn, CgA, Ki-67, P53 were positive. Conclusions Hepatoid adenocarcinoma of the stomach coexisting with neuroendocrine tumor is very rare, it is made up of three structures: hepatoid cell differentiation, adenocarcinoma differentiation and neuroendocrine differentiation area. Diagnosis relies on immunohistochemical and histological examination. Key words: Stomach neoplasms; Neuroendocrine tumors; Pathology, clinical; Prognosis

Objective To explore the significance of changes of matrix metalloproteinase-9(MMP-9), interleukin-6(IL-6), and tumor necrosis factor-alpha(TNF-α) protein level in the cerebrospinal fluid (CSF) of children with viral encephalitis(VE). Methods The concentration of neuron-specific enolase(NSE), structural proteins 100B (S100B), and MMP-9, IL-6, TNF-α in the CSF of VE children were detected by an enzyme linked immunosorbent assay, and the correlations of them were analyzed. Results NSE, S100B, MMP-9, IL-6, TNF-α protein expression could be found significantly higher than those in the control group, and there were significant differences according to statistics expression trends(all P<0.05). The NSE protein expression was significantly positive related with S100B in the VE group (r=0.467, P=0.009), and the concentration was markedly negative related with the duration of viral encephalitis (r=-0.472, P=0.008). MMP-9, IL-6 protein expression were significantly positive related with NSE, S100B respectively (r=0.698, P=0.00; r=0.559, P=0.00; r=0.812, P=0.00; r=0.664, P=0.00). TNF-α protein expression was positive related with CSF S100B(r=0.363, P=0.049), but there was no correlation between TNF-α and NSE(r=0.245, P=0.193). Conclusions The neurons and the neuroglial cells are damaged in the viral encephalitis children.MMP-9, IL-6, TNF-α protein may participate in the pathological damage process of nerve cells in VE children in different degrees. Key words: Viral encephalitis; Structural proteins 100B; Matrix metalloproteinase-9; Interleukin-6; Tumor necrosis factor-alpha; Child

<h3>Objective</h3> To investigate the possible association between 5-lipoxygenase activating protein (ALOX5AP) gene SG13S114T/A polymorphism and acute coronary syndrome (ACS) in the elderly (age≥60 years). <h3>Methods</h3> A case-control study was conducted in 377 aged patients with ACS (male: 242 cases, female: 135 cases, from 60 to 99 (70.24±6.99) year old) (ACS group) and 337 control subjects (male: 167 cases, female: 170 cases, from 60 to 88 (68.72±6.55) year old) who were free from coronary heart disease (control group). The ALOX5AP SG13S114T/A polymorphism was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. <h3>Results</h3> (1) Genotype frequencies of ALOX5AP gene SG13S114T/A AA, AT and TT were 13.79%, 50.93% and 35.28% in patients with ACS and 12.76%, 38.58% and 48.66% in control subjects. There was statistical difference of frequencies of AT and TT genotype between ACS group and control group (p&lt;0.05). The T allele frequency was 69.74% in patients with ACS and 61.22% in control subjects and not significantly different between patients with ACS and controls (p&gt;0.05). (2) Subgroup analysis found that as compared with the control group, the AT genotype frequency in male ACS group was significantly higher (40.72% vs 52.48%, p&lt;0.05) and the TT genotype frequency in female ACS group was significantly lower (61.76% vs 37.04%, p&lt;0.05). Subgroup multivariate logistic regression analysis showed AT and TT genotype and T allele with male ACS as well (p were 0.014, 0.005 and 0.020, respectively). <h3>Conclusion</h3> The AT and TT genotype and The T allele of ALOX5AP gene SG13S114T/A polymorphism may be associated with the susceptibility to ACS in the elderly, especially the aged male, in the Chinese Han population of Sunan region.

The molecular chaperone, heat shock protein gp96 (HSP-gp96), has been shown to have roles in the synthesis, processing and transport of tumor antigens. Therefore, the capacity for HSP-gp96 to induce dendritic cells (DCs), thymus-dependent lymphocytes (T lymphocytes), natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) was investigated. Recombinant adenovirus (AD) containing HSP-gp96 (AD-gp96), as well as gp96-peptide complex from the human leukemia cell lines, K562, HL-60 and U937, was prepared. Purified gp96-peptide complex was found to stimulate the proliferation of T lymphocytes, increase the activity of NK cells and CTLs and induce the secretion of cytokines, compared with ADgp96. In the latter case, levels of IFN-γ and TNF-α were found to increase and levels of IL-12(P70) and IL- 10 decreased. In combination, these results indicated that the gp96-peptide complexes derived from the tumor cells contributed to the activation of lymphocytes and increase the presentation of tumor antigen. Furthermore, the chaperone function of gp96 promoted the maturation of DCs, enhanced the antigen presention function of DCs and induced the secretion of cytokines by DCs. Therefore, gp96-peptide complex derived from the tumor cells potentially represents an immunization therapy for the elimination of residual leukemia cells. Key words: Leukemia, heat shock protein gp96, dendritic cells, cytotoxic T lymphocytes.

Objective To evaluate the impact of loss of heterozygosity (LOH) at chromosome 9p21 and P16INK4A(CDKN2A) expression on the prognosis of gastrointestinal stromal tumor (GIST). Methods A total of 51 cases with GISTs were characterized by immunohistochemistry and evaluated for LOH at 9p21 by microsatellite analysis in 4 markers(D9S1751, D9S1846, D9S942 and D9S1748). Associations of LOH at 9p21 and P16INK4A expression encoded by CDKN2A with clinicopathological parameters and prognosis in GISTs were analyzed. Results The frequency of 9p21 LOH was 37.0% (10/27) at D9S1751, 37.5%(12/32) at D9S1846, 42.1%(16/38) at D9S942 and 24.2%(8/33) at D9S1748. The overall frequency of LOH at 9p21 was 63.3%(31/49). In 21 samples of 51 GISTs(41.2%), P16 expression was not detected. Loss of P16 expression was 60%(12/20) in high risk group and 23.5%(4/17) in very low and low risk groups(P＜0.05). The 5-year overall survival rate of p16-negative patients was 70.8%, while in P16-positive patients it was 92.0%(P＜0.05). Conclusions LOH at 9p21 is a frequent event in GIST. Loss of CDKN2A gene at 9p21 may contribute to the progression and malignant transformation of GIST. P16 expression in GIST is associated with prognosis. Key words: Gastrointestinal stromal tumors;  Loss of heterozygosity;  Gene,P16;  Prognosis

Objective To evaluate two different histopathological classification systems (Fletcher and Miettinen) for the risk in cases of gastrointestinal stromal tumors (GIST). Methods One hundred and sixty-five GIST cases with complete clinicopathologic and follow-up data were evaluated for their biologic potential by the histopathological classification systems of Fletcher, and among those, 164 cases GIST were evaluated by the histopathological classification systems of Miettinen. The implication of two classification systems were compared by survival analysis. Results Evaluated by Fletcher histopathological classification system, 59 cases (35. 8%) were graded as high risk, 49 cases (29. 7%) as intermediate risk, 43 cases (26. 1%) as low risk and 14 cases (8. 5%) were very-low risk. Evaluated by Miettinen's system, 68 cases (41.5%) were as high risk, 23 cases (14. 0%) were intermedatie risk, 60 cases (36. 6%) were low risk and 13 cases (7. 9%) were very-low risk. Evaluated by both two systems, the survival time and disease-free survival time of high risk GIST were lower than those of very-low, low and intermediate risk GIST(P 0. 05). Conclusions Fletcher histopathological classification system is simple and easy to use, while Miettinen's system for evaluating biological potential by anatomic site is more accurate and predictive in the selection of high risk patients for target adjuvant treatment. Key words: Gastrointestinal stromal tumors;  Risk assessment;  Prognosis

In this study, the authors investigated the association between apolipoprotein E (ApoE C112R/R158C) single nucleotide polymorphisms (which determine the E2, E3, and E4 isoforms) and age-related macular degeneration (AMD), and the mechanism underlying the association. Genomic DNA was extracted from 133 clinically screened controls, 94 volunteers with a younger mean age, 120 patients with advanced AMD, and 40 archived ocular AMD slides for single nucleotide polymorphism typing. The effects of recombinant ApoE isoforms on CCL2 (a chemokine), CX3CR1 (a chemokine receptor), and VEGF (a cytokine) expression in cultured human retinal pigment epithelium (RPE) cells were tested and serum cholesterol profiles of the clinically screened subjects were analyzed. ApoE112R (E4) distribution differed significantly between AMD patients and controls. ApoE112R allele frequency was 10.9% in the AMD group when compared with 16.5% in the younger controls and 18.8% in the clinically screened controls. The pathologically diagnosed archived AMD cases had the lowest allele frequency of 5%. No significant differences in ApoE158C (E2) distribution were observed among the groups. A meta-analysis of eight cohorts including 4,289 subjects showed a strong association between AMD and 112R, but not 158C. In vitro studies found that recombinant ApoE suppresses CCL2 and VEGF expression in RPE cells. However, the E4 isoform showed more suppression than E3 in both cases. These results further confirm the association between ApoE112R and a decreased risk of AMD development. The underlying mechanisms may involve differential regulation of both CCL2 and VEGF by the ApoE isoforms.—Hans E. Grossniklaus

风为百病之长，百病之始也，致病广泛，历代医家对'风'的认识有所不同，但始终不离外风与内风之说。随着现今外界环境的变化，风邪伤人较前更甚，临床上对风邪的认识亦逐渐深入。'风'邪可有狭义与广义之分，狭义之'风'单指六淫中的风邪，广义之'风'除了六淫中的风邪外，还包括了其他具有轻扬开泄、善动不居等特点的一切'外风'以及因体内阳气亢逆变动而产生的'内风'。文章从广义之'风'入手，结合小儿生理病理特点，阐述'风邪致病'在儿科疾病发病中的重要性，以期为儿科临床辨证提供参考。

Objective: To assess the clinical value for the clinicopathological features of microcystic elongated and fragmented (MELF) invasion in endometrial carcinoma (EEC) . Methods: The clinicopathological data of 108 cases of endometrial carcinoma with total hysterectomy, bilateral adnexectomy, and pelvic dissection were retrospectively analysis in Peking University People's Hospital from April 2015 to October 2016. Twenty-five patients with endometrial carcinoma showing MELF invasion pattern were collected. We analyzed retrospectively the association of MELF pattern invasion with clinical pathology data and prognosis of the patients, partial immunohistochemical staining was implemented. MELF invasion was a special invasion pattern and characterized by microcystic, elongated, fragmented (composed of cluster cells) gland in muscular layer. Results: The incidence rate was 23.1% (25/108). These patients mean age was (59.3±10.9) years old. Four cases were premenopausal, and 21 were postmenopausal. Abnormal vaginal bleeding was the main clinical presentation. The lesions tend to appear adjacent to the tumor body. Sometimes, it may be appears away from the tumor body in the deep muscle layer.Lymph node metastasis were present in 5 cases (20%, 5/25). Thirteen cases (52%, 13/25) of them demonstrated lymph vascular space involvement (LVSI). The immunohischemical expression of ER,PR, Ki-67 and galectin-3 showing MELF invasion pattern were weaker than no showing MELF invasion pattern endometrial carcinoma, cktokeratin (CK) was showed diffuse strong positive expression, E-cadherin was moderately positive expression. All 25 cases were followed up for (23.2±5.9) months (14-33 months) after the therapy with no recurrence on metastasis. Conclusions: MELF invasion pattern is a special invasion pattern in low-grade EEC. The incidence of LVSI and lymph node metastasis rate in endometrial carcinoma with MELF invasion are significantly increased. The prognosis of MELF invasion pattern may be poor.目的： 探讨子宫内膜样癌伴微囊、拉长及碎片状（MELF）浸润患者的临床病理特点及预后。 方法： 收集2015年4月—2016年10月于北京大学人民医院行子宫全切除+双侧附件切除+盆腔（或加腹主动脉旁）淋巴清扫术的108例子宫内膜样癌患者的临床病理资料，筛选出其中伴MELF浸润的子宫内膜样癌患者25例，回顾性分析其临床病理资料，总结其免疫组化染色的特点，并对其预后进行随访。MELF浸润是指低级别子宫内膜样癌的肌层中以微囊型腺体、拉长变形的腺体以及碎片状腺体形式出现的1种特殊的浸润方式。 结果： 子宫内膜样癌伴MELF浸润的发生率为23.1%（25/108），患者年龄为（59.3±10.9）岁（46~76岁）；绝经前4例、绝经后21例；临床表现以阴道不规则流血为主；MELF浸润灶通常位于紧邻肿瘤主体的周围组织或深肌层中。25例伴MELF浸润的子宫内膜样癌患者的淋巴脉管间隙浸润（LVSI）阳性率（52%，13/25）和淋巴结转移阳性率（20%，5/25）均明显高于不伴MELF浸润的子宫内膜样癌患者[均为2%（2/83），P<0.01]。免疫组化法检测显示，MELF浸润组织中，ER、PR、细胞增殖相关核抗原（Ki-67）、β半乳糖凝集素3（galectin-3）呈弱阳性表达，上皮型钙黏蛋白（E-cadherin）呈中等强度阳性表达，细胞角蛋白（CK）呈弥漫性强阳性表达。25例伴MELF浸润的子宫内膜样癌患者术后随访（23.2±5.9）个月（14~33个月），均未出现复发及转移。 结论： MELF浸润是发生于低级别子宫内膜样癌的特殊浸润方式，伴MELF浸润的子宫内膜样癌患者LVSI阳性率及淋巴结转移阳性率明显增高，提示其预后较差。.

Based on the panel data of meteorological research institutions from 2013 to 2016, this paper uses the BCC model and Malmquist index method (in the DEA model) to analyze the comprehensive efficiency and total factor productivity changes of research and development (R&amp;D) in the meteorological research service industry. This can provide an empirical basis for improving the efficiency of R&amp;D in the meteorological research service industry.

This study tries to evaluate and analyze the competitiveness of meteorological S&amp;T in 31 provinces in China by using difference analysis method, with considering indexes IMD S&amp;T input and output and the global innovation index algorithm, and suggestions for policy-making are presented. These policies try to set up congenial mechanism, bridge the gap among different regions, avoid duplication of research, promote collaboration between various scientific and technological organizations, establish meteorological research infrastructures,open science data and instruments sources to different organizations, and raise public awareness of meteorology.

Pseudomonas aeruginosa est un pathogene opportuniste responsable d'infections graves chez les personnes immunodeprimees, les grands brules et les patients atteints de la mucoviscidose. Cette pathogenicite repose sur de nombreux facteurs de virulence dont le systeme de secretion de type III (SSTT). Nous avons observe une proteine precedemment identifiee, PsrA, necessaire pour la pleine activation de l'expression du SSTT chez P. Aeruginosa. Les analyses par retard de migration electrophoretique de fragments du promoteur de l'operon regulateur exsCEBA ont montre que la proteine recombinante PsrA pourrait se fixer sur celui-ci. Le mutant DpsrA a montre une diminution marquee de la secretion des effecteurs de type III et une faible resistance a la bactericidie par des cellules de type phagocytaires, PLB-985. L'ensemble des resultats suggerent que PsrA est un nouvel activateur qui est implique dans l'expression du SSTT en augmentant le niveau de la transcription d'exsCEBA. Dans un second temps, nous avons mis en evidence qu'un signal inhibiteur, de type quorum sensing inconnu et produit dans la phase stationnaire de la culture, peut reprimer l'expression du SSTT in vitro. L'analyse de milliers de mutants de transposition a montre que la production de ce signal depend du tryptophane, qui est le precurseur de nombreux metabolites dont l'acide d'indole-3-acetic (IAA). IAA-Na et un autre membre de cette famille de molecules, le acide 1-naphthalenacetique (NAA-K) aux concentrations millimolaires peuvent en effet inhiber l'expression et la secretion du SSTT. L'identification precise de ce signal necessite des investigations plus poussees.

Pseudomonas aeruginosa is an opportunistic pathogen that causes acute and chronic infections in hospitalized individuals, burn victims, and cystic fibrosis patients. One of the virulence factors most strongly correlated with severe infection both in animal models and in human patients is the type III secretion system (TTSS).We have observed a previously identified protein, PsrA, necessary for full activation of TTSS gene expression in P. aeruginosa. Electrophoretic mobility shift assays showed that recombinant PsrA could bind to the exsCEBA promoter region. Mutant with a deletion in the psrA gene showed a marked decrease in the secretion of the type III effectors and weak resistance to phagocyte-like PLB-985 cells. Taken altogether, these results suggest that PsrA is a newly identified activator that is involved in the expression of the TTSS by enhancing the exsCEBA transcriptional level.In the second time, we found that an inhibitory signal, which is similar but not known quorum sensing signal, produced in the stationary phase culture supernatant, can repress in vitro the expression of TTSS. The analysis of thousands of transposon insertion mutants showed that the production of this signal might depend on the tryptophan, which is the precursor of many metabolites, such as indole-3-acetic acid (IAA). IAA sodium salt (IAA-Na) and one of IAA-like compound, 1-naphthaleneacetic acid potassium salt (NAA-K) can indeed inhibit the expression and the secretion of the TTSS at millimolar concentrations. The precise identification of this signal requires more investigations.

Uncertainties for predictions of thermal hydraulic problem are caused by the complex physical origin. Uncertainty qualification methods are designed to meet the requirement of regulation to ensure acceptable safety margins with enough confidence. Various methods are proposed for the industrial applications to achieve the proposed level of probability and confidence. However, it is not enough to obtain more statistics information with limited calculation by the full evaluation model. The present paper is based on a simplified model of large break loss of coolant accident for peak clad temperature (PCT) estimation (Catton et al. (1990)). The simplified model is based on physical foundation with sufficient uncertainty parameters and trend indication. The higher PCT during blow down phase and reflooding phase was set to be the target output parameter. The numerous parameters and distribution in the simplified model were set to the source of input uncertainty. The effects of PIRT, sample number, uncertainty distribution were studied. The results provided qualitative and quantitative results for uncertainty methods and would be beneficial for the determination of uncertainty qualification methods in engineering design.

To study gene transfer mediated by electric pulse and optimize the parameters of electric pulse in vivo.10 micrograms plasmid pcD2/LacZ was injected into the quadriceps of 220 Kunming mice. One to two minutes after the DNA injection electric pulse with different parameters was given to the injection site. Three days after, activity of beta-galactosidase was measured, and the expression of Laz2 gene in muscle was determined by histochemical staining.The activity of beta-galactosidase in electric pulse group (131.6 U/mg +/- 86.6 U/mg protein) was 30 fold higher than that in direct injection group (4.9 U/mg +/- 1.0 U/mg protein) (P < 0.05). Histochemical analysis of muscles injected with a LacZ expression plasmid also showed that in vivo electric pulse increased both the number of positively stained muscle fibers and the density of staining. When the electric pulse was with the parameters of 200 V/cm, 40 ms, 6 pulses and 1HZ, maximal gene expression was achieved.Electric pulse, with optimal parameters, increases gene expression. Electric pulse makes much more gene expression than mere intramuscular DNA injection.