BACKGROUND: Runx2 plays a central role in osteogenic differentiation, which is of important significance in new bone formation.OBJECTIVE: To observe the effect of Runx2 over-expression on osteogenic differentiation of human umbilical cordblood mesenchymal stem cells.METHODS: Runx2 was generated by RT-PCR and the recombinant adenovirus (pAd-Runx2) was constructed. The viraltiter was determined by air dilution method. After being transfected into human umbilical cord blood mesenchymal stemcells, the green fluorescence was observed under fluorescence microscope. Real-time fluorescent quantitative PCR andwestern blot were used to detect mRNA expression changes of osteogenesis related genes, Runx2, OCN, BMP-2, ALP,in human umbilical cord blood mesenchymal stem cells at 1, 3, 7 and 14 days after transfection.RESULTS AND CONCLUSION: The recombinant adenovirus was successfully constructed and the titer was1.7×1010 pfu/L. After infection by pAd-Runx2, human umbilical cord blood mesenchymal stem cells expressed greenfluorescence protein clearly under the fluorescence microscope. Cells in the transfected group differentiated intoosteoblast-like cells, and those in the control group stayed the same as pre-infected. The expression of Runx2, OCN,BMP-2 and ALP in the transfected group increased over time to some extent, but these changes were not detected in thecontrol group. These findings indicate that Runx2 over-expression can promote the osteogenic differentiation of humanumbilical cord blood mesenchymal stem cells.

Objective To study the effect of ursolic acid (UA) intervention on hepatocyte apoptosis induced by TGF-β1 and its potential mechanism.Methods Primary hepatocytes were extracted from healthy SD rats by in situ perfusion,cultured for 12-24h,then randomly divided into the following groups:blank control group,UA control group (UA 25μmol/L),TGF-β1 group (TGF-β1 2.5ng/ml),UA intervention group (UA 25μmol/L and TGF-β1 2.5ng/ml),DPI intervention group (DPI 0.5μmol/L and TGF-β1 2.5ng/ml).Each group was treated with drugs for corresponding time and their proliferation and apoptosis were detected by flow cytometry,the expression of CD95 (Fas) mRNA was analyzed by RT-qPCR,the expression of protein CD95 and membrane translocation of NADPH oxidase (NOX) subunit p47Phox were analyzed by Western blotting,and the reactive oxygen species (ROS) generation in primary hepatocytes was analyzed with reactive oxygen detection kit.Results UA intervention at 30min before TGF-β1 stimulating hepatocytes markedly reduced hepatocyte apoptosis (63.97 ± 3.19 vs 80.53 ± 1.56,P＜0.01) and promoted hepatocyte proliferation (18.67 ± 1.60 vs 10.83 ± 2.03,P＜0.01).UA intervention notably down-regulated the expressions of CD95 mRNA and protein (1.28 ± 0.15 vs 2.40 ± 0.25,P＜0.01;1.05 ± 0.15 vs 1.37 ± 0.18,P＜0.05),restrained membrane translocation of p47phox (1.13 ± 0.12 vs 1.76 ± 0.22,P＜0.01),and decreased ROS level in primary hepatocytes induced by TGF-β1 (2.12 ± 0.45 vs 3.23 ± 0.53,P＜0.01).Conclusion The mechanism of UA inhibiting hepatocyte apoptosis induced by TGF-β1 is likely to be that UA intervention reduced hepatocyte apoptosis by inhibiting NOX activation and decrease generation of ROS so as to down-regulate expression of CD95 in hepatocytes.

Objective To investigate the factors for predicting the efficacy of third-generation cephalosporin (3 rd GC) in the treatment of community-acquired spontaneous bacterial peritonitis (CASBP), and to establish and validate an efficacy predictive model for 3 rd GC in the treatment of CASBP. Methods A retrospective analysis was performed for the clinical data of the patients with liver cirrhosis and CASBP who received 3 rd GC monotherapy for initial treatment in The Ninth Hospital of Nanchang, and according to their treatment outcome, they were divided into non-response group and response group. The patients hospitalized from 2013 to 2018 were included in the modeling cohort (55 patients the non-response group and 110 in the response group), and those hospitalized from 2019 to 2020 were included in the validation cohort (17 patients in the non-response group and 43 in the response group). In the modeling cohort, the two groups were compared in terms of the indices including general information, underlying diseases, past history, clinical manifestation, and laboratory test results. Univariate analyses (the t -test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between groups) and a binary Logistic regression analysis were used to identify efficacy predictors, and an efficacy predictive model was established based on the logistic regression equation. The receiver operating characteristic (ROC) curve was plotted to perform internal and external validations of the model in the modeling cohort and the validation cohort, respectively. Results The study population had a mean age of 51.6±12.0 years, and male patients accounted for 80.0%; hepatitis B was the main cause of liver cirrhosis (66.7%), and 3 rd GC had an overall response rate of 68.0%. In the modeling cohort, compared with the response group, the non-response group had significantly lower proportion of patients with the first onset of SBP, polymorphonuclear (PMN) count in ascites, and leukocyte count in ascites (all P < 0.05), as well as significantly higher proportion of patients with exposure to broad-spectrum antibiotic and platelet count (both P < 0.05). The Logistic regression analysis showed that the first onset of SBP (odds ratio [ OR ]=0.158, 95% confidence interval [ CI ]: 0.064-0.392, P < 0.001), ascites PMN count ( OR =0.728, 95% CI : 0.530-0.998, P =0.046), exposure to broad-spectrum antibiotic ( OR =9.152, 95% CI : 1.513-55.351, P =0.016), and platelet count ( OR =1.012, 95% CI : 1.006-1.019, P < 0.001) were independent predictive factors for non-response to 3 rd GC. The efficacy predictive model had an area under the ROC curve (AUC) of 0.840, and based on the maximum Youden index, predictive score ≥ 0.207 was the optimal cut-off value for predicting non-response, with a corresponding Youden index of 0.536, a sensitivity of 89.1%, a specificity of 63.6%, a positive predictive value of 55.1%, and a negative predictive value of 92.1%. This model had an AUC of 0.837 in the validation cohort. Conclusion The first onset of SBP and higher ascites PMN count are the protective factors against non-response to 3 rd GC for the treatment of CASBP, and exposure to broad-spectrum antibiotic and higher platelet count are the risk factors for such non-response. The model established for predicting the efficacy of 3 rd GC in the treatment of CASBP has good predictive performance and thus holds promise for clinical application.