Enhanced expression of TLR4 in HepG2 cells after transient and stable HBV transfection / 中国医师杂志

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.

HBV genotypes and gene mutations in patients with liver failure after withdrawal of lamivudine / 中华临床感染病杂志

Objective To investigate HBV genotypes and gene mutations in chrenic hepatitis B (CHB) patients with liver failure after lamivudine withdrawal. Methods Twenty four patients with relapsing CHB after lamivudine withdrawal were divided into liver failure group ( n = 12 ) and chronic hepatitis group ( n = 12 ). HBV DNA from these patients was amplified by PCR. The PCR products were cloned into PGEM-T vector and HBV DNA sequences were analyzed. Results In liver failure group, there were 6 sequences detected, in which 3 were of genotype B and 3 were of genotype C. In chronic hepatitis group, there were 9 sequences detected, in which 2 were of genotype B and 7 were of genotype C. Compared with the wild type HBV sequences, there were multiples mutations in S, P, C, X regions. Gene mutations in high conservative sequences of BCP and P regions were detected in liver failure patients after lamivudine withdrawal. Conclusions In HBV patients with liver failure after lamivudine withdrawal, half of them were of genotype B and the others were of genotype C. Some mutations in high conservative sequences of BCP and P regions may be related to the liver failure in these patients.

Correlation of Pathology and Serology of Fibrosis in Patients with Viral Hepatitis / 中山大学学报(医学科学版)

【Objective】 To study the relationship of pathologic changes and serologic markers of fibrosis in patients with viral hepatitis. 【Me thods】 Liver s pecimens were obtained by percutaneous needle biopsy under color Doppler ultras ound guidance in 299 patients with viral hepatitis. The specimens were stained b y hematoxylin and eosin (HE), Gordon and Sweet's reticulum methods (RT), in orde r to determine the degree and the stage of pathologic changes with microscopy. H yaluronic acid(HA), collagen type Ⅳ(Ⅳ-C) and human precollagen type Ⅲ（HPCⅢ ）as serum fibrous markers were detected by radioimmunoassay. 【Results】 The se rum levels serologic markers were slightly increased in 97 patients with mild ch ronic hepatitis, moderately increased in 126 patients with moderate chronic hepa titis, and significantly increased in 29 severe cases and 47 subjects with cirrh osis. Both the grade of inflammatory activity and the stage of fibrosis were clo sely related to the levels of serum fibrous markers. 【Conclusion】 Chronic vira l hepatitis pathologic feature and levels of serum markers of fibrosis change al ong with clinical process of patients. The combination of liver biopsy and detec tion of serum markers of fibrosis might be highly valuable for the diagnosis.

Construction of c-myb Antisense RNA Recombinant Retroviral Vector and Establishment of Its Packaging Cell Line PA317 / 中山大学学报(医学科学版)

【Objective】To construct the c-myb antisense RNA recombinant retroviral vector and its packaging cell line.【Methods】The segment of c-myb gene was cloned into pUC19 with TA cloning method after amplication by RT-PCR,and then was subcloned into retroviral vector pDOR.The recombinant retroviral vector named pDOR-myb was transfected into retroviral package cell line PA317 after selection with G418.【Results】Sequencing data indicated that the c-myb gene was exactly identical to the sequence in the GenBank.The segment of c-myb gene was inserted directionally into pDOR.Resistant colonies were obtained and the titers of pDOR-myb were 5.2×104～9.5×104 CFU/mL.【Conclusion】The recombinant retroviral vector containing c-myb gene is successfully constructed and its packaging cell line PA317/pDOR-myb was established.

Expression of TLR2 and TLR4 in hepatocellular carcinoma / 中国病理生理杂志

AIM:To investigate the expression of TLR2 and TLR4 in hepatocellular carcinoma(HCC),and to analyze their correlations to clinicopathologic features of HCC.METHODS:The protein and mRNA levels of TLR2 and TLR4 in HCC and para-tumor tissue were determined by immunohistochemistry and real-time fluorescence quantitative PCR(RFQ-PCR).RESULTS:The protein and mRNA levels of TLR2 and TLR4 in HCC were lower than those in para-tumor tissue(P

Effects of c-myb antisense RNA on the proliferation and collagen Ⅰ gene expression in cultured hepatic stellate cells / 中国病理生理杂志

98%. In the pDOR-myb infected HSCs, c-myb expression levels, the cell proliferation, and ? 1-Ⅰ collagen mRNA expression were repressed significantly. CONCLUSIONS: c-myb plays a key role in the activation and proliferation of HSC. c-myb antisense RNA can inhibit cell proliferation and ? 1-Ⅰ collagen mRNA expression in the infected HSC. These data suggest that inhibition of c-myb gene expression would be a potential way for the treatment of liver fibrosis.