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ObjectiveTo investigate the effects of splenectomy on the intestine mucosa barrier in rats with obstructive jaundice. Methods50 Wistar rats were divided randomly into the obstructive jaundice group (OJ), in which the animals underwent operation to ligate common bile duct, and the obstructive jaundice + splenectomy group (OJ+ S). Seven days post-operation, plasma endotoxin levels were detected. Intestinal mucosa permeability was measured by the ratios of lactulose and mannitol (L/M). Immunohistochemistry and Western blot were used to examine the expression of tight junction proteins (ZO-1 and occludin) in the distal ileum mucosa. Western blots images were analyzed quantitatively. ResultsAverage ratios of L/M and plasma endotoxin were decreased obviously in the OJ+S group compared to those in the OJ group (all P=0. 001). Compared with the OJ group, the average intestinal villus height and mucosa thickness were upgraded somewhat in the OJ + S group (P = 0.019, 0. 001 ). By immunohistochemistry staining seven days post-operation, same comment as above the amounts of strong positive expression of ZO-1 were significantly decreased in the OJ group (6/18, P-0. 021). There wewas no difference between the OJ+S group(8/17) and the OJ group.The amount of strong positive expression of occludin was higher in the OJ + S group than that of the OJ group(10/17 vs 4/18, P= 0. 026). The same outcomes were obtained by quantitative Western blot images. Conclusion The intestinal epithelial permeability was increased in rats with obstructive jaundice,and intestinal barrier was damaged. After excising spleen, the amount and distribution of tight junction proteins were changed and the impairment of intestinal barrier was abated.
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Objective To investigate the correlation of insulin resistance (IR) and benign prostate hyperplasia (BPH). Methods The 200 health examination men were divided into three groups according to the prostate volume (PV). There were 100 healthy subjects as control group (PV≤20 ml), 50 cases in BPH1 group (20 ml＜PV＜50 ml) and 50 cases in BPH2 group (PV≥50 ml).The fasting blood glucose (FBG) and fasting serum insulin (FSI) were determined using hexokinase method and radioimmunoassy, respectively. The IR index (IRI) and body mass index (BMI) were calculated according to the formula in HOMA model. Results There were no statistical differences between BPHl group and healthy control group in IRI (1.10±0. 18 vs. 1.18±0.21) and BMI (22. 0±3.0vs. 21.8±2.7) (t=0.74, 0.18, both P＞0. 05), but the IRI (1.31±0.19) and BMI (24. 8±3.29) increased in BPH2 group (P=0. 01, 0.03). The percentage of hyperglycosemia was higher in BPH patients than in normal controls (25% vs. 5%, P = 0. 00). Of them, the percentage of hyperglycosemia was the highest in BPH2 group (36%, P = 0. 01 ), their IRI ( 1.47 ± 0.21 ) was higher than in euglycemia patients (t=3.92, P=0.00), but the BM1 was unchanged compared with the control group ( 25.8 ± 4.3 vs. 24.3 ± 2.71, P = 0. 95 ). Conclusions There is a positive correlation of IR and hyperglycosemia with severe BPH, and the IR in severe BPH is independent of BMI.
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Objective To investigate the effects and mechanisms of bile on small intestine mucosal barrier.Methods Fifty Wistar rats were assinged into 3 groups randomly: obstructive jaundice(OJ) group(n=20),biliary external drainage group(n=20) and control group(n=10).Ten days after operation,the plasma endotoxin level was determinated,the terminal ileum mucosas was obtained to be morphologically measured by light microscope,and immunohistochemistry and Western blot were uesd to examine the expressions of tight junction proteins zona occludens-1(ZO-1) and occludin in the mucosas.Results Atrophy significantly appeared in the distal ileum mucosas in OJ group.Compared with control group,the intestinal villus height,mucosa thickness and crypt depth in OJ group were obviously decreased 27.8%,21.7%,and 25.4%(P=0.001,0.001,0.040).There were no differences between external drainage group and control group(P=0.050,0.070,0.080);While the values of external drainage group were significantly higher than those in OJ group(all P=0.001).The level of plasma endotoxin was up to(1.49?0.27) EU/ml in OJ group compared with control group((0.27?0.09) EU/ml),P=0.001.In external drainage group,the value was(0.91?0.25) EU/ml,which was obviously higher than that in control group and lower than that in OJ group(all P=0.001).Immunohistochemical study showed strong positive expression of ZO-1 dropped from 7/10 in the control group to 6/20 in OJ group(P=0.040),occludin expression was 8/10 in control group and 7/20 in OJ Group(P=0.020);expressions of them in external drainage group(8/20(P=0.100,0.210) and 9/20(P=0.060,0.200)) displayed no significant differences compared with the other twogroups.Quantitative testing of Western blot showed the expressions of ZO-1 and occludin in OJ group were significantly lower than those in control group(P=0.001,0.010),the values in external drainage group were higher than those in OJ group(P=0.005,0.014).The expression of ZO-1 was lower in external drainage group than that in control group(P=0.001),and there was no significant difference of occludin between the two groups(P=0.062).Conclusion Lack of intestinal bile will undermine the intestinal tight junction protein composition,and make intestinal mucosal barrier impaired.The intestinal barrier more severely injured when biliary tract obstructs because of multiple factors.Bile plays an important role in the maintenance of intestinal mucosal barrier.
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Objective To investigate exogenous PTEN gene transfected human breast cancer cell line MDA MB 468. MethodsUsing the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA MB 468.After transfection, the cells were selected by G418.Then resistant clones were chosen and expanded in DMEM culture medium. RT PCR, immunohistochemical method and western blot were used to determine the expression of target genes. ResultsAn anti G418 cell clone was established and expanded in culture. The transfected PTEN gene MDA MB 468 cells showed expression of PTEN mRNA and PTEN protein. ConclusionHuman breast cancer cell line MDA MB 468 established in this study expresses consistently exogenous PTEN genes.