Objective To observe the expression of TLR4 in hepatocellular carcinoma cell line HepG2 after transient and stable HBV genome transfection. Methods Immunofluorescence flow cytometry was used to detect mean fluorescence intensity (MFI) of TLR4 and TLR4 positive cell percentage in hepetocellular carcinoma cell lines HepG2 and HepG2. 2.15. Various doses of HBV DNA plasmid were transfected into HepG2 cells with lipefectamine 2000. Immunofluoroscenee flow cytometry was used to detect MFI of TLR4 and TLR4 positive ceil rate of infected HepG2 ceils. Trypan blue staining was used to examine the sum of living cells. Results MFI of TLR4 and TLR4 positive cell rate of HepG2.2.15 cells were significantly higher than those in HepG2 cells (both P' <0. Ol). MFI of TLR4 and TLR4 positive cell rate of HepG2 cells transfected by various doses of HBV DNA were significantly higher than those in control group (all P' < 0. 01). MFI of TLR4 and TLR4 positive cell rate of infected HepG2 cells were positively correlated with the doses of HBV DNA (both P' <0. 01) and negatively correlated with the sum of living cells (both P' <0. 01). Conclusions Enhanced expression of TLR4 appeared in HepG2 cells with both transient and stable HBV infection, along with reduction of living cells.

Objective To compare the double-guide wire technique (DGT) with the standard cannulation technique (SCT) in patients with difficult access due to biliary complications after liver transplantation.Methods Difficult CBD cannulation is characterized by unsuccessful cannulation in 10 minutes.A total of 91 patients with biliary complications after liver transplantation were assigned to the DGT group (44patients,including 6 difficult cannulation,41 males and 3 females,30 to 61 years) and the SCT group (47patients,41 males and 6 females,33 to 56 years).An extra 20-minute cannulation was performed on the two groups.Success rate,procedure time and complications were compared.Results CBD cannulation was successful in 36 (81.8％) patients of DGT group and 33 (70.2％ ) patients of SCT group,which was not different ( P ＞ 0.05 ).The time of successful CBD cannulation in the DGT group ( 11.7 ± 3.2 minutes) was shorter than that in the SGT group ( 16.8 ±2.8 minutes,P ＜0.05).The incidence of post-ERCP hyperamylnsemia had no difference in the two groups ( P ＞ 0.05 ).There were no serious complications like infection,hemorrhage or perforation in either group.Mild pancreatitis occurred in 2 cases in the SCT group,but none in DGT.Conclusion DGT is an effective and safe technique in patients with biliary complications after liver transplantation,with no more complications than the SCT group.It is recommended in difficult cannulation of common bile duct (CBD).

ObjectiveTo investigate the changes and role of β-arrestin 1 in the course of non-alcoholic fatty liver disease (NAFLD) progressing to hepatocellular carcinoma (HCC). MethodsEighty healthy male C57BL/6 mice were randomized into the vegetarian diet group and the high fat diet group according to the random number table with 40 mice in each group. Mice in the vegetarian diet group were fed with vegetarian diet (13% calories in fat) and mice in the high fat diet group were fed with high fat diet (58% calories in fat). Eight mice in each group were decapitated at the end of 9 and 24 weeks. The rest mice in each group were decapitated at the end of 48 weeks. The incidence of HCC of two groups was observed. The expression of proteinβ-arrestin 1 in the liver tissues of mice was detected by Western blot and the mRNA level was examined using TaqMan real time fluorescence quantitative RT-PCR. The incidence of HCC in two groups was compared using Fisher's exact test, and the protein β-arrestin 1 expression and mRNA level of two groups were compared usingt test. Spearman correlation analysis was used to evaluate the relationship between protein β-arrestin 1 expression, mRNA level and the feeding duration of high fat diet in high fat diet group.ResultsThe incidence of HCC in the high fat diet group was 18% (4/22), which was significantly higher than 0 (0/23) in the vegetarian diet group (P=0.034). The expression level of protein β-arrestin 1 in liver tissues of mice in the high fat diet group was 2.4±0.5 in the 9th week, which was significantly higher than 1.5±0.4 in the vegetarian diet group (t=2.779,P<0.05). The β-arrestin 1 mRNA level in liver tissues of mice in the high fat diet group in the 9th, 24th and 48th week were 4.1±0.8, 7.8±2.1 and 12.5±1.2 respectively, which were all significantly higher than 2.6±0.7, 3.6±0.6 and 6.9±1.2 in the vegetarian diet group (t=4.029, 5.522, 9.487;P<0.05) . The protein β-arrestin 1 and mRNA level in HCC tissues of mice in the high fat diet group in the 48th week were 4.6±0.5 and 22.0±3.2, which were signiifcantly higher than 1.6±0.4 and 12.5±1.2 in liver tissues at the same period (t=9.600, 7.837;P<0.05). The protein β-arrestin 1 and mRNA level in high fat diet group were positively correlated with the duration of high fat diet (r=0.949, 0.922;P<0.05). Conclusions It is likely to develop NALFD for the mice fed with high fat diet, and the incidence of HCC is signiifcantly increased. β-arrestin 1 may play a role of accelerating the course of NAFLD progressing to HCC.