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Objective To investigate the expression of PRR11 in bladder cancer tissues and its effect on proliferation and apoptosis of bladder cancer cell line T24. Methods The expression of PRR11 was detected using immunohistochemistry method in 57 specimens of bladder urothelial carcinoma and adjacent tissues. The correlations of PRR11 expression with the clinicopathological characteristics of patients with bladder urothelial carcinoma were analyzed. The mRNA and protein expression levels of PRR11 in human immortalized bladder epithelial cell lines SV-HUC-1 and human bladder cancer cell lines HTB-9, T24, J82 and UM-UC-3 were measured by qRT-PCR and Western blot. The gene expression of PRR11 in T24 cells was silenced by lentivirus shRNA. The mRNA expression level of PRR11 was detected by qRT-PCR. CCK-8 was used to detect cell proliferative activity. Cell clonality was detected by plate cloning assays. The rate of apoptosis was evaluated using flow cytometry. The protein expression levels of PRR11, Caspase-3, Bcl-2 and Bax were assessed by Western blot. Results PRR11 was highly expressed in bladder urothelial carcinoma, and its expression level was correlated with the pathological grade and T stage of the tumor. The mRNA and protein expression levels of PRR11 in HTB-9, T24, J82 and UM-UC-3 cells were higher than those in SV-HUC-1 cells (P < 0.05), especially in T24 cells. PRR11 gene silence reduced the expression levels of PRR11 mRNA and protein, as well as the cell proliferation activity and cell clonality, elevated the apoptosis rate, up-regulated the protein expression levels of Cleaved caspase-3 and Bax, and down-regulated the protein expression level of Bcl-2. Conclusion The expression of PRR11 is upregulated in bladder urothelial carcinoma tissues and bladder cancer cell lines. Interfering with PRR11 expression can inhibit the proliferation and promote the apoptosis of bladder cancer T24 cells.
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ObjectiveTo investigate the differentially expressed proteins in the plasma exosome of acute-on-chronic liver failure (ACLF) patients with different prognoses, to analyze their functions and biological processes, and to provide a basis for clinical diagnosis. MethodsA prospective study was performed for 10 ACLF patients who were hospitalized and diagnosed in Beijing YouAn Hospital, Capital Medical University, from July 2019 to October 2019, and the patients were followed up for 90 days. The patients who died or received liver transplantation were enrolled as liver transplantation/death group (5 patients), and the patients who survived were enrolled as survival group (5 patients). The Mann-Whitney U test was used for comparison of general data between the two groups. The label-free quantitative proteomic method was used for identification and quantitative analysis of plasma exosome proteins to screen out differentially expressed proteins, and a functional enrichment analysis was performed. R-3.5.1 software was used to perform a hierarchical cluster analysis of differentially expressed proteins to analyze the biological processes involving these proteins. ResultsA total of 860 proteins were identified by the exosome proteomic analysis, and according to the criteria of upregulation ＞1.2 folds or downregulation ＞1.2 folds (P＜0.05), there were 116 differentially expressed proteins. Compared with the liver transplantation/death group, the survival group had 62 upregulated proteins and 54 downregulated proteins. The bioinformatics analysis showed that these differentially expressed proteins mainly participated in immune reaction, signal transduction, vesicle-mediated transport, cell death, and cell proliferation and were closely associated with the signaling pathways including inflammatory response, carbohydrate and amino acid metabolism, hepatocyte injury, and hepatocyte regeneration. ConclusionDifferentially expressed proteins screened out by the label-free quantitative proteomic method can be used as serological markers for the early diagnosis and prognostic evaluation of ACLF.
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Objective@#To investigate the role of the glycogen synthase kinase 3β (GSK3β) and the peroxisome proliferator-activated receptor alpha (PPARα) signaling pathway in acute liver failure and related mechanisms in a mouse model of acute liver failure induced by D-galactosamine/lipopolysaccharide (D-GalN/LPS).@*Methods@#C57BL/6 mice were given intraperitoneal injection of D-GalN/LPS to establish a mouse model of acute liver failure. SB216763 was used to inhibit the activity of GSK3β and PPARα siRNA was used to inhibit the expression of PPARα. Western blotting was used to measure the expression of PPARα protein. The changes in liver pathology were observed to evaluate liver injury, and the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured to assess liver function. Quantitative real-time PCR was used to measure the mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-12p40 (IL-12p40), and PPARα. A one-way analysis of variance was used for comparison of means between multiple groups; the least significant difference test was used for data with homogeneity of variance, and the Games-Howell method was used for data with heterogeneity of variance.@*Results@#In the mice with liver failure induced by D-GalN/LPS, GSK3β inhibition promoted the mRNA and protein expression of PPARα (F = 13.18 and 301.36, P = 0.00 and 0.00). In the mice with acute liver failure induced by D-GalN/LPS, GSK3β inhibition alleviated liver bleeding, inflammation, and necrosis and reduced the serum levels of ALT (F = 25.16, P = 0.000) and AST (F = 12.96, P = 0.001), as well as the mRNA expression of TNF-α (F = 32.17, P = 0.00), IL-1β (F = 11.57, P = 0.005), and IL-12p40 (F = 14.17, P = 0.015) in liver tissue. The inhibition of PPARα expression reversed the liver-protecting effect of GSK3β inhibition, which manifested as aggravation in liver bleeding, inflammation, and necrosis, increases in the serum levels of ALT (F = 25.16, P = 0.001) and AST (F = 12.96, P = 0.000), and an increase in the mRNA expression of TNF-α (F = 32.17, P = 0.00), IL-1β (F = 11.57, P = 0.024), and IL-12p40 (F = 14.17, P = 0.001) in liver tissue.@*Conclusion@#In mice with acute liver failure induced by D-GalN/LPS, the GSK3β-PPARα-inflammatory factor signaling pathway may play an important role. GSK3β inhibition has a protective effect in mice with acute liver failure possibly by activating the inhibitory inflammatory factor of PPARα.
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BACKGROUND: Intravenous hypnotics are used in pregnancy, labor and delivery. The aim of the present study was to investigate and compare the relaxant effects of propofol, thiopental, ketamine, and etomidate on isolated rat uterine smooth muscles. METHODS: Uterine smooth muscle preparations were obtained from non-pregnant female rats. The uterus of the rat was dissected and cut into 10 mm strips. The muscle strips were bathed in Krebs solution. After spontaneous uterine contractile activity had been accomplished, propofol, ketamine, thiopental, and etomidate in various concentrations were added cumulatively to the baths and resting tension, active tension, and frequency of contration were recorded at each concentration of agents. EC(5), EC(25), EC(50), EC(75), and EC(95) of each drug on active tension and frequency of contraction were calculated using a probit model. RESULTS: Propofol, thiopental, and etomidate reduced uterine contractions in a concentration-dependent manner. Ketamine concentrations of 10(-7) to 10(-5) M augmented uterine contractions but ketamine concentrations of 10(-4) to 10(-3) M attenuated uterine contractions. The EC(50)'s of propofol, thiopental, ketamine, and etomidate on active tension were 1.56 x 10(-5) M, 4.97 x 10(-5) M, 3.52 x 10(-4) M, and 2.73 x 10(-5) M, respectively. CONCLUSIONS: All four intravenous hypnotics relaxed the uterine smooth muscle of rats except for ketamine in low concentrations (10(-7) to 10(-5) M). Propofol had the greatest relaxant effects on isolated rat uterine smooth muscle among these hypnotics. It seems that ketamine is a suitable obstetric hypnotic agent for hypovolemic parturients and propofol is a useful hypnotic agent for uterine relaxation during pregnancy.