Objective To solve the shortage of hepatocytes for l iver tissue engineering, to explore the possibil ity of prol iferation of rat bone marrow mesenchymal stem cells (BMSCs) and the feasibil ity of differentiation of BMSCs into hepatocyteswith a culture system containing cholestatic rat serum and hepatocyte growth factor (HGF) in vitro. Methods Myeloid cellsof femur and tibia were collected from the female healthy Wistar rats at the age of 6 weeks, the BMSCs were isolated, purified and identified. Normal and cholestatic rat serum were prepared from 40 healthy Wistar rats at the age of 12-14 weeks. The 3rd passage of BMSCs were harvested and added different cultures according to the following grouping: group A, DMEM plus 10%FBS; group B, hepatocyte growth medium (HGM) plus 5%FBS; group C, HGM plus 5% normal rat serum; group D, HGM plus 5% cholestatic rat serum; group E, HGM plus 5% cholestatic rat serum plus 25 μg/L HGF. The changes of cell morphology were observed, MTT assay was used to measure cell growth; the expression of alpha-fetoprotein (AFP) and cytokeratin 18 (CK18) were detected by immunocytochemistry; the glycogen deposit was examined by periodic acid-schiff (PAS) staining; and the urea content in culture supernatant was determined by glutamate dehydrogenase. Results Polygonal cells and binuclear cells were observed in groups D and E, while the shapes of cells in groups A, B, and C did not obviously change. The cell growth curve demonstrated that the speed of cells proliferation in group C was the fastest, the one in group B was the slowest; showing significant differences when compared with groups A, D, and E (P lt; 0.05). On the 7th day in groups D and E, the positive expressions of AFP and CK18 emerged, on the 14th day the positive expression of glycogen emerged. At the same period, the expression ratio was higherin group E than in group D (P lt; 0.05). The urea concentration increased gradually with induction time in groups D and E, the concentration was higher in group E than in group D (P lt; 0.05). No expressions of AFP, CK18, glycogen, and change of the urea concentration were observed in groups A, B, and C. Conclusion Normal rat serum can obviously promote the growth of BMSCs; cholestatic rat serum which promote the growth of BMSCs can induce to differentiate into hepatocyte; and a combination of cholestatic serum and HGF can increase the differentiation ratio.
Objective To evaluate the efficacy and safety of endoscopic variceal ligation (EVL) versus endoscopic variceal sclerotherapy (EVS) for acute esophageal variceal bleeding in patients with liver cirrhosis.Methods We searched CBMdisc (1979 to 2006), CNKI (1994 to 2006) and VIP for randomized controlled trials (RCTs) and quasi-RCTs comparing EVL and EVS for acute esophageal variceal bleeding patients with liver cirrhosis. The methodogical quality of included trials was critically assessed and the data were extracted by two reviewers, working independently. The Cochrane Collaboration’s RevMan 4.2.7 software was used for meta-analysis. Results Nine RCTs involving a total of 1371 patients were included: 688 in EVL group and 683 in EVS. The meta-analyses showed a significant reduction for mortality [RR 0.60, 95%CI (0.36, 0.98)], and non-significant reductions in complications, rebleeding and emergency hemostasis in the EVL group compared to the EVS group. EVS was non-significantly better than EVL for the rate of eradication varices and recurrent varices. Conclusions For acute esophageal variceal bleeding in patients with liver cirrhosis, EVL has better effect and fewer complications than EVS. However, because the quality of included RCTs was poor, the strength of our conclusions was limited. Further high-quality RCTs are required.
【摘要】 目的 探讨丙型肝炎病毒非结构蛋白NS4B对肝细胞内p53表达的影响，以及在肝癌发生中的作用与机制。 方法 设置空白对照组、空白载体组、转染NS4B组、转染p53组、共转染NS4B及p53组。使用脂质体介导转染法，转染丙型肝炎病毒非结构蛋白重组质粒PCXN2-NS4B及突变型p53基因重组质粒pC53-CX22AN3进入Chang肝细胞内，并用G418筛选获得稳定表达细胞。采用免疫细胞化学法检测p53表达率。 结果 空白对照组无p53表达，空白载体组及转染NS4B组呈弱阳性表达，转染p53组及共转染组呈阳性表达；转染p53组、共转染组分别与空白对照组、空白载体组及转染NS4B组比较，差异均有统计学意义 （Plt;0.05）。 结论 NS4B可能抑制p53表达，也可能阻止其进入细胞核，但NS4B与突变型p53关系不明确。NS4B导致肝细胞异常增生，诱导肝癌发生可能不依赖p53的异常表达及突变。【Abstract】 Objective To investigate the effect of hepatitis C Virus on-structural protein 4B(HCV NS4B) on expression of p53 in hepatic cell, and to study the role and mechanism in development of hepatocellular carcinoma. Methods The experiment was divided into negative control, pure vector PCXN2, PCXN2-NS4B, PC53-cx22AN3, and co-transfection group. Recombinant plasmid PCXN2-NS4B and mutant p53 gene--PC53-cx22AN3, PC53-cx22AN3 with PCXN2-NS4B, blank vectors were transfected into Chang liver cell by liposome-mediated transfection respectively. Positive cells were screened by G418. The expression rate of p53 was measured by immunocytochemistry. Result No expression rate of p53 gene in control group was found, lower positive expression in group PCXN2 and PCXN2-NS4B. The expression of p53 gene in group PC53-CX22AN3 and co-transfection was ber than the others (Plt;0.005). Conclusion HCV-NS4B may inhibit the expression of p53 gene, and it may play a crucial role in inhibiting p53 transfered to hepatic cells nuclear. But it isn’t clear that the. HCV-NS4B can enhance the role of mutant p53 gene. It suggested that HCV-NS4B induce proliferation of hepatic cell not through regulating the expression of p53.