AIM: To investigate the role of Smad7 in the Smad2 expression induced by transforming growth factor-?_1(TGF-?_1) in rat peritoneal mesothelial cells(PMCs).METHODS: Rat PMCs were cultured at different doses of TGF-?_1 (0,1.25,2.5,10 ?g/L) for different time(0,5,15,30,60,120 min).PCDNA3-Smad7 was then transfected into cultured rat PMCs by lipofectamine,and the cells were stimulated like the above.Endogenous Smad2 and Smad7 expression was evaluated by RT-PCR and Western blotting.RESULTS: TGF-?_1 induced increase in Smad2 mRNA and protein expression at 5 min,peaked at 30 min,and declined to baseline levels at 120 min, which was in a time-dependent manner.TGF-?_1 also induced Smad7 mRNA expression at 5 min,and then declined,down to the lowest at 30 min,but at 60 min it increased again.Smad2,Smad7 mRNA and protein expression induced by TGF-?_1 were also dose-dependent.After transfection,overexpressions of Smad7 mRNA and protein in rat PMCs were observed,which did not decline with time.The expression of Smad2 mRNA significantly decreased by 33%,56%,67%,71%,63% and 57%(P

AIM: To investigate the effects of advanced glycation end products on activation of Smad signaling pathway and collagenⅠ synthesis in proximal tubular epithelial cells.METHODS: Advanced glycation end products(AGE-BSA) were prepared by incubation of bovine serum albumin(BSA) with D-glucose.Normal rat proximal tubular epithelial(NRK52E) cells were cultured in RPMI-1640 medium with AGE-BSA.Phosphorylation and nuclear translocation of Smad2/3 were examined by immunocytochemistry.Levels of TGF-?_1 in supernatant of cell culture were measured by enzyme-linked immunosorbent assay(ELISA).Expression of TGF-?_1,Smad2,Smad3 and Smad7 mRNA were detected by RT-PCR.Expression of ?-SMA,E-cadherin and collagenⅠproteins were detected by Western blotting.RESULTS: AGE-BSA induced Smad2/3 phosphorylation and nuclear translocation,two peaks occured at 30 min(68% vs 16%,P

AIM: To observe the effect of antibiotic treatment on the inflammatory mediator expression in peritoneum and the peritoneal transport function in rats with acute peritonitis, and explore its mechanisms. METHODS: Eighty-six SD rats were randomly divided into three groups. Control group (n=28) were treated with PBS (ip), peritonitis group (n=28) and treatment group (n=28) were challenged with the E.coli (ip), but at 3 h and 9 h gentamicin was given (ip) in treatment group. Seven rats of every group were randomly sacrificed at 24 h, 48 h, 72 h and 7 d. Peritoneal equilibration test (PET) was did before they were killed. Leukocyte count, pathological changes and the expression of CD45, NF-?B, IL-1?, TNF-? in peritoneum were examined. RESULTS: (1)The blood leukocytes in peritonitis group decreased strikingly, but did not change obviously in other two groups. The peritoneal fluid leukocytes in peritonitis group increased significantly from 24 h to 72 h, while in treatment group, it enhanced more strikingly than peritonitis group at 24 h, and recovered earlier. (2) Both in peritonitis group and treatment group, the expression of activated NF-?B, IL-1?, TNF-? and CD45 increased significantly, but the treatment group was lower than model group at 48 h and 72 h. The mRNA level of IL-1? and TNF-? had the same trend as their protein expression. (3) Compared with the control group, UF and D/D_0 Glu decreased significantly in model group and treatment group, and D/PTP increased dramatically. The D/P TP in treatment group lowered obviously compared with peritonitis group, while the net UF and D/D_0 Glu had not significant difference between treatment group and model group. CONCLUSION: Antibiotic treatment can partly decrease the expression of inflammatory mediators in peritoneum of rats with acute peritonitis and also can improve the protein transport ability to some extent, but can not improve the peritoneal ultrafiltration and the glucose transport function.

Objective:To analyze the clinical phenotype and molecular pathogenesis of nine patients with hereditary factor Ⅴ (FⅤ) deficiency.Methods:Nine patients with hereditary FⅤ deficiency who were admitted to the Institute of Hematology and Blood Diseases Hospital from April 1999 to September 2019 were analyzed. The activated partial thromboplastin time, prothrombin time, and FⅤ procoagulant activity (FⅤ∶C) were measured for phenotypic diagnosis. High-throughput sequencing was employed for the F5 gene mutation screening, Sanger sequencing was adopted to confirm candidate variants and parental carrying status, Swiss-model was used for three-dimensional structure analysis, and ClustalX v.2.1 was used for homologous analysis.Results:The FⅤ∶C of the nine patients ranged from 0.1 to 10.6. Among them, eight had a hemorrhage history, with kin/mucosal bleeding as the most common symptom (three cases, 37.5%) , whereas one case had no bleeding symptom. There were five homozygotes and four compound heterozygotes. A total of 12 pathogenic or likely pathogenic mutations were detected, of which c.6100C>A/p.Pro2034Thr, c.6575T>C/p.Phe2192Ser, c.1600_1601delinsTG/p. Gln534*, c.4713C>A/p.Tyr1571*, and c.952+5G>C were reported for the first time.Conclusion:The newly discovered gene mutations enriched the F5 gene mutation spectrum associated with hereditary FⅤ deficiency. High-throughput sequencing could be an effective method to detect F5 gene mutations.