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Aim: To characterize high mobility group box chromosomal protein 1 (HMGB1) polymorphisms in patients infected with hepatitis B virus (HBV) and determine the different patterns in patient subgroups., Methods: A total of 1495 unrelated Han Chinese HBV carriers were recruited in this hospital-based case-control study. The HMGB1 1176 G/C polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism assay., Results: A significant association was observed between HMGB1 1176 G/C polymorphism and outcome of HBV infection. The subjects bearing 1176G/G genotype had an increased risk of susceptibility to chronic hepatitis B, liver cirrhosis and severe hepatitis B when compared with those bearing at least one 1176C allele., Conclusion: Patients with 1176G/G genotype of HMGB1 gene are more likely to have a progressive status in HBV infection.

The organic anion transporting polypeptide 1B1 (OATP1B1, encoded by SLCO1B1) plays an important role in the transport of endogenous and xenobiotic compounds, such as bile acids and rifampin. In this study, the association between OATP1B1 polymorphisms and rifampin hepatotoxicity was investigated using integrated population genetic analysis and functional studies. A total of 273 unrelated patients treated with rifampin were recruited. The allele frequencies were examined in patients with drug (rifampin)-induced liver injury (DILI) (n = 118) and without (non-DILI) (n = 155). Functional analyses were conducted to determine whether the inhibition of bile acids by rifampin was associated with OATP1B1 variants. In the present study, 24 single nucleotide polymorphisms (SNPs) in OATP1B1 were detected in a Chinese population, with two of them causing an amino acid change (rs2306283 and rs4149056). The haplotypes constructed by these two SNPs were OATP1B1 *1a, *1b, *5 and *15, with their respective frequencies being 23.44, 66.30, 0.73 and 9.52% in a total of 273 individuals. The logistic regression analysis indicated that the *15 haplotype was associated with susceptibility to DILI (p = 0.03, OR = 2.04, 95% CI 1.05-3.96). The frequency of the *15 haplotype in DILI patients was significantly higher than that in non-DILI patients (p = 0.03). In the subgroup analysis, the *15 haplotype was associated with susceptibility to cholestatic/mixed injury (p = 0.03, OR = 2.31, 95% CI 1.06-5.02). Functional assessment of the OATP1B1 *15 haplotype revealed that the activity of bile acid uptake was markedly reduced compared to the three other haplotypes. In the inhibition study, the inhibition by rifampin in the *15 haplotype was greater compared to that in the other haplotypes. These results suggest that the OATP1B1 *15 haplotype is an important predisposing factor for rifampin-induced liver injury.

Aim: To assess the rigorous relationship between human leukocyte antigens (HLA)-DR alleles and outcomes of hepatitis B virus (HBV) infections by means of meta-analysis., Methods: Medline/PubMed, EMBASE, CNKI and VIP were searched to identify relevant studies. Study quality was evaluated using the Newcastle-Ottawa Scale. Odds ratios (OR) and 95% confidence interval (95% CI) were pooled using Stata 11.0. Subgroup analyses were performed by ethnicity. Heterogeneity and publication bias analyses were performed to validate the credibility., Results: A total of 2609 patients with chronic hepatitis B and 2606 controls spontaneously recovering from prior HBV infection were included. Meta-analysis showed that HLA-DR*04 (OR = 0.72, 95% CI: 0.60-0.85) and DR*13 (OR = 0.27, 95% CI: 0.19-0.37) alleles were significantly associated with HBV clearance while patients carrying HLA-DR*03 (OR = 1.47, 95% CI: 1.16-1.87) or DR*07 (OR = 1.59, 95% CI: 1.24-2.03) alleles had a significantly increased risk of chronic HBV persistence. For the HLA-DR*01 polymorphism, a significantly association with HBV clearance was found in Chinese Han group (OR = 0.48, 95% CI: 0.26-0.86), but not found in other ethnic groups (P = 0.191). For other polymorphisms, no association with the HBV infection outcome was found., Conclusion: HLA-DR*04 and DR*13 alleles may be the protective factors for HBV clearance and HLA-DR*03, and DR*07 alleles may be the risk factors for HBV persistence.

Objective: To investigate the dynamic correlation between pre-S1 antigen, pre-S2 antigen and HBV DNA in the serum of chronic hepatitis B (CHB) patients undergoing nucleoside analogue therapy., Methods: 12 CHB patients with transient virological response after lamivudine treatment, and 20 patients treated with adefovir for 5 years were recruited in this study. Serum samples were collected at four time points when HBV DNA fluctuated sharply during lamivudine treatment, and at 0, 8, 12, 28, 52, 104, 156, 208, 260 weeks following adefovir treatment. HBV DNA was quantified by real-time PCR, pre-S1 and pre-S2 antigens were detected by ELISA., Results: The titers of pre-S1 and pre-S2 antigens were not correlated with the HBV DNA level in the serum of lamivudine treated patients. Only in one case of the adfovir treated patients, the decrease of pre-S1 and pre-S2 antigens was in parallel with the decrease of HBV DNA. Linear regression analysis indicated that neither pre-S1 antigen nor pre-S2 antigen was correlated with HBV DNA in the serum of lamivudine or adfovir treated patients (P more than 0.05)., Conclusion: Our results indicate that the titers of pre-S1 and pre-S2 antigens are not correlated with the serum HBV DNA in CHB patients undergoing nucleoside analogue therapy. Neither pre-S1 nor pre-S2 is a good predictor for the outcome of nucleoside analogue treatment.

Objective: Studies have shown that potassium channel plays a pivotal role in T cell activation. The expression of potassium channel gene KCTD9 was evidenced being highly upregulated in patients with severe hepatitis B (SHB). To understand this phenomenon further, tissue and cellular expression profiles of KCTD9 were investigated in patients with SHB., Methods: A rabbit peptide polyclonal antibody was prepared. Various samples including peripheral blood mononuclear cells (PBMCs); livers from patients with SHB or mild chronic hepatitis B, were examined for KCTD9 expression by quantitative real time PCR and immunohistochemistry staining (IHC). Confocal microscopy was used to illustrate the localizations of the expressions., Results: Increased expression of KCTD9 was observed in PBMC in over 35.7% of the patients with SHB when compared with that of patients with mild chronic hepatitis B. In all patients, the relative value of increased KCTD9 mRNA was positively correlated with alanine aminotransferase, aspartate aminotransferase, total bilirubin and direct bilirubin but negatively with serum albumin. The expression was mainly located in hepatocytes, bile duct epithelial cells, Kupffer cells and inflammatory cells, and in the cytoplasm of PBMCs from the healthy individuals and patients with mild chronic hepatitis B, whereas in both cytoplasm and nuclei in those from patients with SHB., Conclusion: The increased expression of potassium channel gene KCTD9 correlates with disease severity in patients with viral hepatitis B.

Objective: To study genotype distribution and the characteristics of hepatitis B virus (HBV) in Uighur patients with chronic hepatitis B (CHB) in Xinjiang, China., Methods: Type specific primers and PCR were used to detect the HBV genotypes of 127 Uighur CHB patients in Xinjiang. Genotyping results were confirmed by PCR product sequencing., Results: Among the 127 patients, the proportions of genotype D, B, C and B/D, C/D, B/C/D were 39.4% (50/127), 22.0% (28/127), 16.5% (21/127) and 9.4% (12/127), 8.7% (11/127) and 3.9% (5/127), respectively. The distribution of the HBV genotypes showed no significant differences between male and female patients (x2 = 8.058, P > 0.05), between HBeAg positive and negative patients (x2 = 6.033, P > 0.05), and between patients of different ages (x2 = 3.137, P > 0.05)., Conclusion: Genotype D HBV is predominant in Uighur patients with chronic hepatitis B in Xinjiang. The distribution of various HBV genotypes shows no significant differences between these Uighur patients with different HBeAg positivity, sex and age.

Objective: To investigate whether human L02 hepatocytes could survive after implanting them into normal, immunocompetent rats., Methods: Human L02 hepatocytes were injected through the uterine walls into the intraperitoneal cavities of fetal Sprague-Dawley rats to induce immune tolerance to human L02 hepatocytes. Human L02 hepatocytes stained with DiI were implanted into the spleens of the 2-week old rats. Immuno-fluorescent staining, SP immunohistochemistry, and DiI staining were used to detect human albumin and specific proliferating cell nuclear antigen (PCNA) in the rat livers. The distribution of human L02 hepatocytes was observed under the fluorescent microscope., Results: Dynamic distribution of human L02 hepatocytes in the rat livers was observed from the 1st to the 10th week after the implantation. Human albumin was detected at 2, 4, 6 and 8 weeks, and at the 4th week it had the highest level. Specific human PCNA was detected in the rat livers from the 2nd to the 6th week after implantation. The PCNA positive cells were most abundant at the 4th week., Conclusion: Human L02 hepatocytes can survive and proliferate for 10 weeks after implanting them into genetically normal immunocompetent rats.

Objectives: To investigate the quasispecies dynamics of hepatitis B virus (HBV) during the course of exacerbation and resolution of chronic hepatitis B in a patients., Methods: Five serum samples were collected from a patient with two episodes of exacerbation and resolution of chronic hepatitis B. A method of PCR-TA cloning-conformation sensitive gel electrophoresis (CSGE)-sequencing was performed to study the dynamic changes of HBV quasispecies in basal core promoter (BCP), precore and core regions of HBV genome., Results: Quasispecies complexity was 10 and 12 at the points of exacerbations, and 14 and 17 at the points of resolutions (t = 3.133, P < 0.05). Ratio of dominant quasispecies in HBV population was high (42.4% and 51.5%) during exacerbations and low (30.3% and 21.2%) during resolutions (t = 3.295, P < 0.05). All dominant quasispecies, except the one during the second resolution, carried core P5T, L60V, S155T, and precore G1896A mutations., Conclusion: The composition of HBV quasispecies changes due to the change of host immune status, and immune pressure might lead to the selection of immune escape mutants.

Objective: To analyze prognostic factors of and to develop a prognostic model for patients with chronic severe hepatitis (CSH)., Methods: From December 1998 to October 2003, 385 in-patients being treated for chronic severe hepatitis were evaluated. The main clinical and laboratory variables were analyzed as predictive factors of survival with Cox univariate and multivariate regression models., Results: The median survival time of this study group was 47 days. The survival rates at 1, 3, 6 months, 1-year, and 3 years were 66.2%, 32.9%, 26.9%, 22.9% and 17.7% respectively. Four prognostic factors were extracted using Cox's proportional hazard model; the prognostic index (PI) was calculated using the following formula consisting of these factors. PI = 0.016loge + 1.148 hepatic encephalopathy + 0.294loge (bilirubin micromol/L) - 0.826loge (prothrombin time activity). The model accurately predicted 3 months survival in an independent series of 84 patients with chronic severe hepatitis., Conclusion: The developed model is valuable in prognostic evaluation of chronic severe hepatitis and it may be useful to guide clinicians in selecting treatment methods for CSH.

Objective: To analyse the difference of genome-wide DNA methylation status of CpG island between the monozygotic twins with disconcordant phenotype of HBV infection and to discover possible differentially methylated genes., Methods: Modified AIMS (amplification of inter-methylated sites) method was adopted. According to the frequent sites of CpG islands (-CGCG- and -CCGG-), three groups of isochizomers with distinct methylation sensitivity (Sma I-Xma I, Hpa II-Msp I and BssH II-Pau I/BseP I) were used to modify the AIMS method. The modified AIMS method combined with Personal Molecular Imager FX system analysis and radioautographic analysis was used to make the global detection of the methylome of CpG islands. Multiple anonymous bands were compared between the twins with the Quantity One bio-soft. The different bands were cloned into T vectors and positive clones were sequenced. BLAST analysis of positive clone sequences was conducted to give the clues for the differential methylated genes between monozygotic twins with discordant phenotype of HBV infection., Results: Nearly the same bands were found between one pair of twins with concordant phenotype of HBV infection. Different methylated bands were found not only between monozygotic twins with concordant phenotype but also between those with disconcordant phenotype. More differential methylated bands were found in the latter groups. By BLAST analysis with sequences of differential methylated bands, four possible genes were got. These genes might relate to the monozygotic twins with disconcordant phenotype of chronic HBV infection., Conclusion: Differential methylation of genes occurs in monozygotic twins with discordant phenotype of HBV infection. Whether these changes are involved in the pathogenesis of different phenotypes needs further elucidation.

Objective: The primary comparative analysis between the host genetic factors and their relationships with clinical phenotype of 20 pairs of HBV infected and high risk twins., Methods: Zygosity of twins was diagnosed by STR microsatellite polymorphism analysis. To identify the serological model and exclude the evidence of coinfection with other virus, we detected HAV, HBV, HCV, HDV, HEV serological markers by electrochemiluminescence method. HBV DNA level was detected by Lightcycler Fluorescent Quantitative system and Liver function (ALT, AST, TBil) was detected by HITACHI7250 Biochemistry Detection System. The data was analysis by Fisher's exact test to comparatively analyze among the monozygotic twins (MZ), dizygotic twins (DZ) and control groups., Results: The significant difference was found in the concordance rate of disease, concordance of clinical phenotype and serological patterns of HBV infection between the MZ and DZ twins (P < 0.05), it was also found between MZ and control groups (P < 0.05), but not between DZ and control groups (P > 0.05). No significant difference in the concordance of other HBV infection markers was observed (P > 0.05). Concordance of clinical phenotype may be related to patients' age and antivirus therapy. Injection with the HBIg and initiative vaccination right after birth can prevent twins with high risk to infection to be victims., Conclusion: The significant difference was found in the concordance rate, concordance of clinical phenotype and serological patterns between MZ and control groups, it was also found between MZ and DZ groups, which is correspond to the opinion of the high concordance of MZ and indicated the host genetic factors may play role in influencing the clinical phenotype, while other factors such as the vaccination may have an effect on the clinical phenotype in some extent.