Objective To establish a cisplatin (DDP)-resistant HepG-2 cell line,and to explore the role of Wnt/β-catenin signaling pathway on multidrug resistance in human hepatocellular carcinoma.Methods The HepG-2 cells were exposed in a gradually increasing dose of DDP to establish a cisplatin ( DDP)-resistant HepG-2 cell line.MTT assay was used to detect the cytotoxic activity of DDP against HepG-2 and HepG-2/DDP cells.The mRNA expression of β-catenin was determined by Real-time PCR assay.The small interfering RNA was used to specifically knockdown β-catenin expression in HepG-2/DDP cells.The protein expression was detected by western blot analysis.Results The DDP-resistant cell line HepG-2/DDP was established by gradient DDP induction successfully.The IC50 values of DDP against HepG-2 and HepG-2/DDP cells were (2.29 ± 0.14) μmol/L and ( 20.51 ± 0.84 ) μmol/L,respectively ( t=95.68,P＜0.01 ),HepG-2/DDP cells was 8.96 times than HepG-2 cells on the resistance of cisplatin.The result of real time PCR showed that 2-△Ct value of β-catenin in HepG-2 cells and HepG-2/DDP cells were (0.323±0.065) and (0.674 ±0.097) (P＜0.01 ).And the protein expression of cisplatin in HepG-2/DDP cells was also significantly higher than that in the HepG-2 cells.The expresssion of β-catenin was significantly and specifically depleted by siRNA duplexes(P＜0.01 ).The IC50 values of cisplatin against HepG-2/DDP cells were (21.02 ± 1.64) μmol/L in cisplatin control group,(6.23 ± 0.68 ) μmol/L in SiRNA targeting interference group and ( 20.44 ± 1.26 ) μmol/L in SiRNA negative interference group,and there was significant difference between control group and SiRNA targeting interference group ( P＜0.01 ).Conclusion The Wnt/β-catenin signaling pathway was activated on the cisplatin(DDP)-resistant HepG-2 cell line and down regulation of β-catenin increased the chemosensitivity of HepG-2/DDP cells against cisplatin.It provided a theoretical basis for finding the new targets of multidrug resistance in liver cancer.

Objective Measuring the therapeutic effect of the isolated duodenal exclusion for type 2 diabetes mellitus rats by observing the post-surgery change of glycometabolism-related indicators which could be contributed to the isolation between food and the duodenal mucosa with an endoluminal sleeve placing into the duodenum intestine duodennal of type 2 diabetes mellitus Rats.Methods In this experiment,12 male type 2 diabetes mellitus Goto-Kakizaki (GK) rats were randomized to endoluminal exclution as the experimental group and 12 to sham surgery as the control group,The body mass,the average daily food intake and fasting plasma glucose were determined before (0 week) and 1,3,6,12 weeks after operation;For testing the hematoglobinA1c(HbA1c) level,blood samples were collected before and 6,12 weeks after operation.Results No statistical significant differences between two groups before operation index.In both groups,The body mass and average daily food intake decreased markedly at 1 week after surgery:the experimental group body mass preoperative(262.6 ± 5.6 g)vs postoperative(224.0 ± 6.3 g) ;the average daily food intake preoperative(25.5 ± 2.7 g) vs postoperative (16.5 ± 3.0 g),P ＜ 0.05.Changes in the experimental group with control group,P ＜0.05.The 3 week,6 week,12 week body weight was gradually increased in rats of two groups.The experimental group daily feed intake is always lower than the preoperative postoperative,P ＜ 0.05.In the 3 weeks,the control group's body weight and daily food intake were higher than those before operation,P ＜ 0.05.The experimental group post-FPG level (7.5 ± 1.1) mmol/L,(7.2 ± 1.2) mmol/L,(7.3 ± 0.9) mmol/L,(7.1 ± 1.0) mmol/L vs preoperation (12.2 ± 1.2) mmol/L were decreased significantly,P ＜ 0.05.Compared to the preoperation,the HbA1c concentration of the experimental group decreased significantly,6 w (7.8 ± 0.9) ％,12 w(8.2 ± 1.2) ％ vs the preoperation (10.3 ± 1.4) ％,P ＜0.05.These indicators of the control group showed no significant changes,P ＞ 0.05.Conclusion The isolated duodenal exclusion can achieve the therapeutic effect for type 2 diabetes mellitus on GK rats,and which might be attributed to the improvement of the glycometabolism.

Objective To explore the effect of the association of low molecular heparin and Galectin-3 on the cell migration and cell proliferation of vascular endothelial cell from mesenchymal stem cells.Methods Depending on the administration, this study was divided into four groups: low molecular weight heparin group, adding 20 mg/L low molecular weight heparin into the cells;Galectin-3 group, adding 5 mg/L Galectin-3 into the cells;combination group, adding 20 mg/L low molecular weight heparin and 5 μg/ml of Galectin-3 into the cells;control group, equal volume of PBS buffer into the cells.The proliferation of vascular endothelial cell was to be detected by EdU incorporation,the cell growth cycle of vascular endothelial cell was to be detected by flow cytometry,and the cell migration of vascular endothelial cell was to be detected by scrath test.Then to investigate the effect of different conditions on the migration and proliferation of vascular endothelial cell.Results The OD490 of MWH group, Galectin-3 group, combined group and control group were (0.285±0.018), (0.297±0.041), (0.351±0.016), and (0.233±0.005) respectively, which indicates that the combined group could increase the cell proliferation significantly(P<0.05).Cultured for 24 hours, the cell migration rate was(42.02±7.62), (45.82±3.96), (68.53±11.22),and (34.21±3.99), suggesting that combined group have the largest cell migration(P<0.05).Conclusion The association of low molecular heparin and Galectin-3 could improve the cell migration and cell proliferation of vascular endothelial cell from esenchymal stemcells significantly.

Objective To explore the diagnosis of abdominal aortic aneurysm(AAA), operative opportunity, the way of operation selecting and the application of blood vessel prosthesis. Method The clinical data of 7 patients with AAA were analysed retrospectively.Results The type of DeBakey I thoracoabdominal aortic aneurysm patient was died of aortic aneurysm broke suddenly before operation, the other 6 patients with AAA underwent aortic aneurysm blooking, opening and replacement of blood vessel prosthesis. Dumbbell thoracoabdominal aortic aneurysm was operated by stayes. No complications of operation occurred. Conclusions Operation is a efficient way in treatment of abdominal aortic aneurysm. Diagnosis timely and definitly, selecting the way of operation in reason and blood vessel prosthesis are the keys to operation, while the correct treatment of perioperation period is the important guaranty of successful therapy.

Objective To determine the effect of prolonged hyperoxia on neonatal rat lung. Methods Full term and premature newborn SD rats were continuosly exposed to 85% oxygen or room air 7 and 14 days after birth.The activities of 3 different kinds of antioxidant enzyme (AOE) including superoxide dismutase(SOD), glutathion peroxidase (GP) and catalase (CAT) in supernatant fractions of lung homogenates were assessed after 7 and 14 days of exposure. So was the lung hydroproline content. Results (1)AOE acctivitis: Except CAT activity at 14 days of exposure, others AOE activities in O 2 exposed rat pups were significantly higher than those in air exposed controls (P

Objective To investigate the role of andrographolide (AP) in protection of carbon tetrachloride (CCl4)-induced acute liver injury in mice and the possible mechanisms. Methods The mice were randomly divided into five groups, including two groups with different doses of AP (50 mg/kg and100 mg/kg), a control group, a CCl4 model group, and a silymarin group. Serum levels of alanine aminotransferase (ALT), aspartateminotransferase (AST), hepatic malondialdehyde (MDA), and glutathione (GSH) were examined. Pathological changes in the liver were observed. RT-PCR was used to detect the expressions of tumor necrosis factor-a (TNF-α) and heme oxygenase-1 (HO-1) mRNA. Results As compared with CCl4 model group, serum levels of ALT and AST and hepatic MDA activity were significantly decreased in AP group (100 mg·kg-1), along with a remarkable increase in hepatic GSH content. Pretreatment with AP at a high dose alleviated histopathological changes induced by CCl4. A markedly increased level of TNF-a induced by CCl4 was reduced by AP, while HO-1at transcriptional level was dramatically elevated following AP pretreatment. Conclusions AP plays a role in protection of CCl4-induced acute liver injury by inhibiting lipid peroxidation and reducing formation of free radicals, the mechanism may be involved in inhibition of TNF-αand activation of HO-1.

Humans ; Liver Transplantation ; methods ; Transplantation, Autologous ; methods

Humans ; Polyomavirus ; classification ; Polyomavirus Infections ; virology

Objective:To investigate the clinical manifestation and determine the distribution of pathogens and their characteristics of drug susceptibility to bloodstream infections (BSIs), and provide evidence for clinical anti-infection treatments after renal transplantation.
Methods:Totally 81 episodes of BSIs occurred in 71 patients between July 2003 and June 2013. We retrospectively analyzed the pathogens and their drug susceptibility characteristics with BD microbiological assay system. We also collected the clinical and laboratory data of the patients . Results:The main pathogens were gram negative bacteria (67.90%), followed by gram positive bacteria (28.40%) and fungi (3.70%). The most common gram negative bacillus was Escherichia coli.While for gram positive bacteria, the main bacillus was coagulase-negative staphylococci. The gram negative bacteria were relatively sensitive to aminoglycosides and carbapenem. The gram positive bacteria were sensitive to glycopeptides and oxazolidone.
Conclusion:The clinical manifestations included high body temperature, onset in the early period after kidney transplantation and high mortality. Though gram positive coccus plays an important role, most infections are caused by gram negative bacteria in BSIs after the renal transplantation. The antibiotic resistant rate for gram negative bacteria is very high as well as gram positive bacteria.

Objective　To explore the expression and significance of p21WAF1 and p53 in HCC. Methods　Immunohistochemical method (IHC) was used to localize and semi-quantitate the proteins of p21WAF1 and p53 and to observe the relationship between the expression of p21WAF1 and the different histopathologic characters in 38 patients of HCC and their peri-cancer tissue as well as 5 normal liver tissue. Results　Of all 38 cases, both p21WAF1 and p53 expression were significantly higher in tumor than that in corresponding non-tumors liver tissue; 14 (36.8 %) of 38 cases showed p21WAF1 positive staining, 28 cases (73.7 %) were p53 positive, p21WAF1＋/p53＋ or p21WAF1-/p53－ were observed in 18, while 20 cases showed p53＋/p21WAF1－ or p53－/p21WAF1＋. p21WAF1＋ was seen in 1 of 38 (2.6 %) corresponding non-cancerous tissue and 2 of 5 normal liver tissue. p53 protein was not detected neither in the non-tumorous tissue nor in normal liver. No significant association was found between the expressions of p21WAF1 and p53(P＞0.05) in HCC. Their was no significant correlation between p21WAF1 or p53 expression and the different histopathologic characters of tumor (differentiating grades, intrahepatic metastasis and/or cancerous thrombi within portal veins). Conclusion　Both p21WAF1 and p53 proteins are over expressed in HCC than that in corresponding non-tumorous liver tissue, but there is no relationship between them. Both p53-independent and p53-dependent mechanism may play a role in regulating p21WAF1 expression in HCC. p21WAF1 immunostaining cannot be used to assess the status of p53 in any given cell or tissue.