Objective To investigate the effects of Celecoxib,a selective COX-2 inhibitor,on the healing of gastric ulcer in rats,and reveal the rale of ERK signal transduction pathway in the mechanism of delaying the healing of gastric ulcer.Methods Forty-eight rats were randomly divided into model group(n=40) and sham operation group(n=8).In the rats in model group acetic acid-induced gastric ulcer was reproduced,while in sham operation group,rats underwent the sham procedure.Eight rats in sham-operation group and eight rats in model group were euthanized three days after the procedure.The other thirty-two rats in model group were divided into two subgroups including Celecoxib group and NS group(n=16).Three days after the procedure,rats in Celecoxib group received a gavage of 0.2% Celecoxib solution,and those in NS group received equal amount of 0.9% NaCl solution.Rats in Celecoxib group and NS group were euthanized on sixth and ninth day after ulcer induction(8 rats at each time point in each group).The effects of Celecoxib on the healing of gastric ulcer were observed.Its effects on the activity of Raf-1 and ERK1/2,and the expression level of two transcription factors c-Fos and c-Jun were also determined by Western blot analysis.Results Nine days after ulcer induction,the ulcer area was 11.9?3.1mm2 and 19.7?3.8mm2 in NS group and Celecoxib group,respectively,and they were much smaller than those on third day(P

Objective To explore the experience in diagnosis and treatment ot primary transitional cell carcinoma of prostate for the early and accurate to diagnosis and treatment.Methods The clinical data and features of 3 cases were retrospectively reviewed.Results All patients were diagnosed as primary transitional cell carcinoma of prostate.Two cases were advanced tumor.The preoperative examinations (ultrasound and serum,PSA) have failed to accurately indicate the diagnosis.All of them were confirmed by pathological examination.1 case lost follow-up,1 case performed TURP + chemotherapy through intravesical administration have survived 17 months respectively till today.The other case has already survived 2 months postoperatively but with lumbar spine bone metastasis.Conclusions Early diagnosis of primary transitional cell carcinoma of prostate is difficult.The diagnosis of the disease depends on the transrectal needle biopsy of the prostate or the specimens of the prostate after the Urethroscopy.Because of the prognosis is bad,and prone to pathological missed diagnosis or misdiagnosis.Need to exclude multicentric lesions and mixed tumor and choice of treatment method.

AIM: To investigate whether Janus kinase/signal transducer and activator of transcription ( JAK/STAT) signaling pathway mediates high glucose-induced damage in human umbilical vein endothelial cells ( HUVECs ) . METHODS:The cell viability was examined by CCK-8 assay.The expression levels of JAK2, STAT3, caspase-9 and en-dothelial nitric oxide synthase ( eNOS) were detected by Western blot .The intracellular levels of reactive oxygen species ( ROS) were tested by DCFH-DA staining followed by photofluorography .Mitochondrial membrane potential ( MMP) was measured by rhodamine 123 staining followed by photofluorography .RESULTS:Treatment of HUVECs with high glucose (40 mmol/L glucose) for 6~12 h enhanced the protein level of phosphorylated JAK 2, peaking at 9 h.Treatment of the cells with high glucose for 6~12 h also increased the protein level of p-STAT3 with the peak value at 12 h.Pretreatment with the inhibitor of JAK/STAT pathway AG490 for 1 h before exposure of the HUVECs to high glucose significantly inhibi-ted the high glucose -induced injury , as evidenced by an increase in the cell viability , decreases in the expression of caspase-9 and the intracellular ROS production , and increases in MMP and the expression of eNOS .CONCLUSION:JAK/STAT signaling pathway is involved in the high glucose-induced damage in HUVECs .

AIM: To explore whether exogenous hydrogen sulfide (H2S) depresses high glucose (HG)-in-duced injury by modulating the Janus kinase/signal transducer and activator of transcription ( JAK/STAT) pathway in hu-man umbilical vein endothelial cells (HUVECs).METHODS:The protein levels of JAK2, STAT3 and cleaved caspase-3 were determined by Western blot.The cell viability was measured by CCK-8 assay.Mitochondrial membrane potential ( MMP) was detected by rhodamine 123 staining followed by photofluorography.The intracellular level of reactive oxygen species (ROS) was analyzed by DCFH-DA staining followed by photofluorography.The activity of superoxide dismutase (SOD) was also measured.RESULTS:Pretreatment of the HUVECs with 400 μmol/L NaHS (a donor of H2S) for 30 min prior to exposure to 40 mmol/L glucose ( HG) markedly attenuated HG-induced upregulation of the phosphorylation of JAK2 and STAT3.Pretreatment with 400μmol/L NaHS for 30 min or with 20μmol/L AG490 (inhibitor of the JAK/STAT pathway) for 30 min attenuated the injury of HUVECs induced by HG, as indicated by the increases in cell viability and SOD activity, and decreases in the protein level of cleaved caspase-3, ROS generation and dissipation of MMP.CONCLU-SION:Exogenous H2 S protects HUVECs against HG-induced injury by inhibiting JAK/STAT pathway.

AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .

Objective To investigate the clinical feature and antibiotic resistance profile of K.pneumoniae isolates from patients for better management of K.pneumoniae infections.Methods Nonduplicate K.pneumoniae strains were collected from January to December in 2015.K.pneumoniae strains were identified by VITEK 2-Compact 60 and tested for antimicrobial susceptibility by KirbyBauer method.Results A total of 753 strains ofK.pneumoniae were included,most (40.9％,308/753) of which were isolated from sputum,followed by urine (18.2％,137/753).Most of the strains were from old patients at least 60 years of age (40.8％,307/753),and primarily from intensive care units (16.7％,126/753) and Department of Respiratory Medicine (13.7％,103/753).Respiratory tract infection was found in 144 patients,of which 71.5％ (103/144) were due to K.pneumoniae.More than half of the K.pneumoniae strains were resistant to piperacillin (66.3 ％),cefazolin (60.8 ％) and cefitroxime (59.4 ％).Only a few strain were resistant to imipenem (2.4 ％) and meropenem (2.0).ESBLs were produced in 410 (54.4 ％) of the 753 strains,and 29 (3.9 ％) strains were carbapenem-resistant,492 (65.3 ％) strains were resistant to multiple antimicrobial agents.Conclusions Clinical K.pneumoniae isolates are highly resistant to most of the antimicrobial agents tested.The strains were mostly isolated from sputum and urine,and positive for ESBLs.MDR K.pneumoniae sWains are emerging.K.pneumoniae isolates are still very susceptible to carbapenems in vitro.

Human tissue typing methods were employed in developing a porcine allotransplantation model. 23 Chinese Sichuan White Pigs(2-3 months old, 17.5+/-4.6kg, with clear family background) were selected for tissue typing, ABO blood type cross reaction, complement-dependent cytotoxicity (CDC) cross reaction and one way mixed lymphocyte reaction (MLR). 6 pairs of swine that showed better matching results were selected as donors and recipients. Single-kidney orthotopic transplantation was conducted after removing both kidneys of the recipient. Five recipients showed low matching results (MLR ranging from 2175 to 3560, CDC from 1 to 4); of them, 2 died of operation, 2 died of acute renal tubular necrosis and accelerating rejection 4 days after operation respectively, and 1 died of acute renal tubular necrosis 4 days after operation. 6 recipients showed high matching results (MLR ranging from 982 to 1916, CDC from 2 to 4); of them, 1 died of anaesthesia during operation, 3 died of accelerating rejection and acute rejection 2 weeks after operation respectively, 1 had good kidney function, and 1 presented weak rejection 1 week after operation but the kidney function came back to normal afterwards. Human tissue typing methods could be adopted in developing the porcine model. Hyperacute rejection could be avoided by screening ABO blood type, CDC and MLR tests. However, based on these primary data, it was hard to evaluate the predictive values of CDC and one-way MLR for accelerating rejection, acute rejection and graft chronic dysfunction. Further research by expanding experiments in these aspects is still going on.
ABO Blood-Group System ; immunology ; Animals ; Cytotoxicity Tests, Immunologic ; Graft Rejection ; prevention & control ; Histocompatibility Testing ; Kidney Transplantation ; immunology ; Swine ; T-Lymphocytes ; immunology ; Transplantation, Homologous ; immunology

Visualizing living cells growing on co-cultured biomaterials is ideal for material biocompatibility evaluation in vitro. In this experiment, mouse fibroblasts L929 were labeled by introducing the gene coding enhanced green fluorescent protein (EGFP) marker into the cells. Morphology as well as proliferation of labeled cells surrounding or on the surface of co-cultured denture base resin slides were observed by use of phase-contrast microscope and fluorescent microscope directly. It was found that residual methyl methacrylate (MMA) in the denture base resin exhibited transient cytotoxicity to fibroblasts and this transient cytotoxicity could be eliminated by pre-extracting the resin with ddH2O for a short time. This fact demonstrated that even slight cytotoxicity of materials could be detected through imaging of living cells near material or material touched. And it was suggested that imaging of living cells co-cultured with biomaterial is helpful to understanding biocompatibility of materials more accurately.
Acrylic Resins ; Animals ; Biocompatible Materials ; Cell Division ; Cells, Cultured ; Culture Media ; Dental Materials ; Denture Bases ; Fibroblasts ; cytology ; Green Fluorescent Proteins ; Indicators and Reagents ; analysis ; Luminescent Proteins ; genetics ; Mice ; Polymethyl Methacrylate ; Transfection

To evaluate the efficiency of three in vitro refolding methods for a humanized single-chain Fv antibody against human CTLA4(CD152) expressed in E. coli, the denatured and purified inclusion bodies (IBS) were refolded by dilution, dialysis and in situ refolding via Immobilized Metal-Ion-Affinity Chromatography (IMAC), respectively. The concentration of refolded scFvs was examined by Bradford method. And the antigen binding activity of the refolded scFvs was analyzed by indirect cell-ELISA. The highest and lowest refolding yields could be obtained by dialysis and in situ refolding via IMAC, respectively. The binding activity of the refolded scFv by dialysis was 1.95-fold higher than that by dilution, 4.13-fold higher than that by in situ refolding via IMAC (GSH/GSSH excluded) and 3.63-fold higher than that by in situ refolding via IMAC (GSH/GSSH included), respectively. In conclusion, a high refolding yield and binding activity of scFv with natural conformation could be obtained by dialysis in the condition of 0. 15 mol/L sodium chloride, 50 mmol/L Tirs-HCl, pH 8. 0 buffer containing 3 mmol/L reduced glutathione and 1 mmol/L oxidized glutathione for 48 hours at 4 degrees C.
Antigens, CD ; biosynthesis ; genetics ; immunology ; Antigens, Differentiation ; biosynthesis ; genetics ; immunology ; CTLA-4 Antigen ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Objective:To investigate the influencing factors of hyperuricemia(HUA) and explore early intervention of metabolic diseases.Methods:A total of 70 523 participants were selected from the database of check-ups in 2016. Univariate analysis and logistic regression analysis were used to identify related factors of HUA. Correspondence analysis was performed for the aggregation of different levels of uric acid(UA) and related factors. The mediating effect of mean blood pressure(MBP) between abnormal metabolic indicators and abnormal renal function was tested.Results:The age, sex, occupation, body mass index(BMI), systolic blood pressure, diastolic blood pressure, blood urea nitrogen(BUN), creatinine(Cr), estimated glomerular filtration rate(eGFR), fasting plasma glucose(FPG), total cholesterol(TC), triacylglycerol(TG), high density lipoprotein-cholesterol(HDL-C), low density lipoprotein-cholesterol, plasma viscosity were significantly related to HUA( P<0.001). Logistic regression analysis showed that youth, male, hypertension, TC, TG, and Cr were risk factors for HUA, while HDL-C was a protective factor for HUA( P<0.001). Correspondence analysis showed that during the gradual increase of UA, TC was the first to appear abnormal, followed by hypertension and TG, and the increase of Cr appeared last. Mediating effect showed that in changes of UA, the mediating effects of MBP on TC, TG, and HDL-C were 36.35%, 12.63%, and 9.41%, respectively. In changes of eGFR, the mediating effects of MBP on TC, TG and HDL-C were 30.20%, 27.70%, and 6.13%, respectively. Conclusions:UA is positively correlated with blood pressure, TC, and TG, and inversely with HDL-C. TC and TG have an impact on renal impairment, in which MBP plays a mediating role.