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).