T3098C and T53C Mutations of HBV Genotype C Is Associated with HBV Infection Progress

Objective To analyze the association between mutation（s） in preS region of HBV and hepatitis B disease progress in Chinese patients with genotype C chronic HBV infection. Methods Ninety-three patients with chronic genotype C HBV infection, including 24 asymptomatic carriers （ASC）, 26 patients with chronic hepatitis B （CHB）, 22 patients with liver cirrhosis （LC） and 21 HCC patients were investigated. Levels of HBV DNA, HBeAg, alanine aminotransferase （ALT）, asparate transaminase （AST） were measured. HBV preS region was analyzed by PCR direct sequencing. Results The prevalence of preS T3098C and T53C mutations ofgenotype C HBV was significantly higher in LC and HCC patients than ASC and CHB patients. The rate ofT3098C mutation in ASC, CHB, LC, and HCC patients were 0.00% （0/24）, 3.85% （1/26）, 9.09% （2/22）, and 30.77% （8/22）, respectively （P=0.0015）, while the rate of T53C mutation was I2.50% （3/24）, 3.85% （1/26）, 40.91% （9/22）, and 42.31% （11/26）, respectively （P=0.0012）. Conclusion The frequency of genotype C HBV preS T3098C and T53C mutations is associated with hepatitis B infection progression.

Phylogenetically clustering of rhizobia by genome structure:application to unclassified Rhizobium

Previous research reveals that the genome structures of rhizobial type strains and reference strains can reflect their phylogenetic relationships. In order to further explore the potential application of genome structure as a phylogenetic marker in rhizobial natural taxonomy, this study analyzed the genome structures of 29 unclassified nodule bacteria isolated from the root nodules of leguminous trees, Robinia sp., Dalbergia spp., and A lbizia spp. and 7 rhizobial reference strains by I-CeuI cleavage, then clustered these bacteria phylogenetically based on their genome structures and compared these clusters with those based on numerical taxonomy and 16S rDNA PCR-RFLP. Eleven phylogenetic clusters were obtained, The clusters were in large part consistent with those based on numerical taxonomy and 16S rDNA PCR-RFLP. Also there are inconsistent clusters based on the above three methods. But results are completely consistent with 16S rRNA clusters. This suggested that the genome structure clustering method can be used to lastly identify root nodule isolates and detect their phylogenetic relationships. The credibility and repeatability of the results, together with the simplicity and possibility to analyze a large number of strains in a short time of the method, indicates the broad potential application of genome structure as phylogenetic marker to categorize rhizobial isolates and should in the future facilitate biodiversity studies.

Analysis of Translocation of the CagA Protein and Induction of a Scattering Phenotype in AGS Cells Infected with Helicobacter pylori

Objective To investigate whether the presence of structured CagA proteins in Western- and Eastern-type Helicobacter pylori （H. pylori） induces different incidences of gastric diseases. Methods CagA and phosphorylatd CagA were expressed in AGS gastric epithelial cells infected with wild type and mutant strains. The ability of individual CagA was determined by immunoprecipitation and Western blot assay. Morphological changes of these cells were observed under microscope to evaluate the appearance of elongation hummingbird phenotype. Results The sizes of CagA proteins in different strains were different, and no phosphorylated CagA proteins were detected in wild-type strains. Meanwhile, the kinetics of CagA status in AGS infected with H. pylori was detected. The molecular weight of phosphorylated CagA with the same size of CagA proteins in H. pylori was different in infections with different wild-type strains. CagA and phosphorylated CagA increased in a time-dependent manner after the infection. The hummingbird phenotype with H. pylori for time-course was observed under microscope. Instead of HPK5 strain, the wild-type 26695 strain induced hummingbird phenotype in a time-dependent manner. Conclusion Translocation and phosphorylation of CagA are necessary, but not sufficient, for the induction of hummingbird phenotype in AGS cells.

Alteration of liver N-glycome in patients with hepatocellular carcinoma

Purpose: Alteration of liver function during pro- gression of hepatocellular carcinoma (HCC) and cirrhosis affects the serum glycoprotein pattern. In this study, the changes in the N-glycome in liver tis- sue from patients with hepatocellular carcinoma and cirrhosis caused by hepatitis B virus infection were investigated to find out the relationship between this maker and liver disease. Methods: Twenty patients, 11 with cirrhosis and 9 with hepatocellular carcinoma, and 15 healthy donors were involved in this study. Liver protein N-glycans were profiled using the DSA-FACE technique developed in our laboratory. To further analyze the fucosylation status of these liver glycans Western lectin blots of total liver proteins were performed using Aspergillus oryzae lectin (AOL) as probe, which is a carbohydrate- binding protein that recognizes specifically α-1,6-fu- cosylated glycans. Results: The N-glycome of liver proteins in patients with HBV related HCC and cirrhosis was analyzed. Compared with healthy donors, the N-glycome had significantly less (p < 0.05) high mannose (M8) in both groups of patients. The total core α-1,6-fucosy-lation in total liver glycoproteins was dramatically increased during the progress of hepatocellular carcinoma and cirrhosis compared to the controls. Conclusion: These results show that fucosylation not only increases in serum proteins but also in liver tissue itself of patients with HBV related HCC and cirrhosis.

Hepatitis B Virus S Promoter Deletion in Hepatocellular Carcinoma

Objective To identify the difference and significance of dominant types of hepatitis B virus(HBV) pre-S mutation between liver tumor tissues and paired adjacent non-tumor tissues and to test if the mutations were tumor tissue specific.Methods HBV DNA isolated from 34 paired intratumoral and peritumoral tissues of hepatocellular carcinoma(HCC) patients were screened by PCR and direct sequencing.All patients carried HBV with genotype C,except for one B/C heterozygote.The expression,localization and excretion of LHBs mutant carrying pre-S deletions were characterized in vitro.The expression of endoplasmic reticulum(ER) GRP78 mRNA was assayed.Results Four patterns of pre-S mutations were identified:pre-S 1 in-frame deletion involving the first start codon;pre-S2 in-frame deletion;pre-S2 start codon mutation with or without in-frame deletion;and S promoter in-frame deletion(ASP).The first two types were evenly found in both tumor and non-tumor tissues.They were rarely present as dominant strains.The last two types were frequently found in the dominant strains in tumor tissues.The overall prevalence of HBV carrying ASP was 17.64%(6/34) in tumor tissues,but none were dominant in nontumor tissues.HBV carrying ASP was unable to produce S protein in vitro.Immunocytofluorescence assay showed that the ASP LHBs mutant aggregated in the cytoplasm,accumulating mainly in the ER.Transient transfection and expression of ASP mutant caused GRP78 up-regulation in vitro.Conclusions HBV S promoter deletion was found dominantly in HCC tumor tissue.The aggregation of mutant large surface proteins in the ER possibly involved in HBV-related HCC.

Management of hepatitis B in China

Hepatitis B virus （HBV） infection is a global public health problem. According to the data of WorldHealth Organization （WHO）, 2 billion people worldwide have been infected with HBV, and among them 350400 million are chronic HBV carriers. Hepatitis B causes about 1 million deaths of HBV related liver failure, cirrhosis, and hepatocellular carcinoma annually.

Cellular stress and redox activity proteins are involved in gastric carcinogenesis associated with Helicobacter pylori infection expressing high levels of thioredoxin-1

Heficobacterpylori infection is related to the development of gastric diseases. Our previous studies showed that high thioredoxin-1 （Trxl） expression in H. pylori can promote gastric carcinogenesis. To explore the underlying molecular mechanisms, we performed an isobaric tags for relative and absolute quantitation （iTRAQ）-based quantitative proteomic analysis of stomach tissues from Mongolian gerbil infected with H. pylori expressing high and low Trxl Differences in the profiles of the expressed proteins were analyzed by bioinformatics and verified using Western blot analysis. We found three candidate proteins, 14-3-3α/β, glutathione-S-transferase （GST）, and heat shock protein 70 （HSP70）, in high Trxl tissues compared with low Trxl tissues and concluded that cellular stress and redox activity- related proteins were involved in the pathogenesis of gastric cancer associated with H. pylori Trxl.

Guideline of Prevention and Treatment for Chronic Hepatitis B (2nd Version)

This guideline is established to standardize the prevention,diagnosis and antiviral therapy of chronic hepatitis B(CHB).For other treatment regimens and methods involving CHB,please refer to relevant guidelines and consensuses.The Chinese Society of Hepatology,Chinese Medical Association(CMA)and the Society of Infectious Diseases,CMA organized relevant native experts to establish this Guideline of Prevention and Treatment for Chronic Hepatitis B(1st version)in 2005,and made the first revision in 2010.In the past 5 years,great progress has been made in the native and foreign fundamental and clinical research with respect to CHB,necessitating additional revision of this guideline.

Vaccination against hepatitis B： the Chinese experience

Objective To review the implementation of mass vaccination of hepatitis B vaccine and its critical role in prevention of hepatitis B virus infection in China.Data sources The data were mainly from PubMed, China Hospital Knowledge Database, and other popular Chinese journals published from 1980 to 2008. The search term was ＂hepatitis B vaccine＂.Study selection Original studies conducted in China and critical reviews authored by principal investigators in the field of hepatology in China were selected.Results Chinese investigators started to develop hepatitis B vaccine in late 1970s. The first home-made plasma-derived vaccine became available in 1986, which has been completely replaced by the domestically produced recombinant （yeast or Chinese hamster ovary cell） vaccine since 2001. China health authority recommended vaccinating all infants in 1992. From then on, China has put tremendous efforts in implementation of mass vaccination. The overall coverage of hepatitis B vaccine in infants has increased steadily and reached more than 95.0% in urban and 83.0%-97.0% in rural areas. The chronic HBV carrier rate in children 〈10 years of age decreased from 10.0% before the mass vaccination to 1.0%-2.0% in 2006, and that in general population decreased from 10.0% to 7.2%; overall, the nationwide mass hepatitis B vaccination has reduced more than 30 million of chronic HBV infections and HBV related severe sequlae.Conclusion The Chinese successful experience in control of hepatitis B by mass vaccination offers an example for any unindustrialized country whoever is committed to control this disease.

Intracellular CMTM2 negatively regulates human immunodeficiency virus type-1 transcription through targeting the transcription factors AP-1 and CREB

Background The CKLF-like MARVEL transmembrane domain-containing family （CMTM） is a novel family of proteins linking chemokines and TM4SF. Different members exhibit diverse biological functions. In this study, the effect of intracellular CMTM2 on regulating human immunodeficiency virus type-1 （HIV-1） transcription was evaluated.Methods The effects of CMTM2 on regulating full-length HIV-1 provirus and the HIV-1 long terminal repeat （LTR）-directed transcription were assessed by luciferase assay. Transcription factor assays, using the luciferase reporter plasmids of AP-1, CRE, and NF-κB were conducted to explore the signaling pathway（s） that may be regulated by CMTM2. The potential relationship between CMTM2 and the transcription factor AP-1 was further analyzed by Western blotting analyses to investigate the effect of CMTM2 on PMA-induced ERK1/2 phosphorylation.Results The results from the current study revealed that CMTM2 acts as a negative regulator of HIV-1 transcription.CMTM2 exerted a suppressive action on both full-length HIV-1 provirus and HIV-1 LTR-directed transcription.Transcription factor assays showed that CMTM2 selectively inhibited basal AP-1 and CREB activity. Co-expression of HIV-1 Tat, a potent AP-1 and CREB activator, can not reverse CMTM2-mediated AP-1 and CREB inhibition, suggesting a potent and specific effect of CMTM2 on negatively regulating these two signaling pathways.Conclusion Intracellular CMTM2 can negatively regulate HIV-1 transcription, at least in part, by targeting the AP-1 and CREB pathways. Exploring the mechanisms further may lead to new ways to control HIV-1 replication.

An unbalanced PD-L1/CD86 ratio in CD14^＋＋CD16^＋ monocytes is correlated with HCV viremia during chronic HCV infection

Circulating monocyte subsets with distinct functions play important roles in hepatitis C virus （HCV） infection. However, the mechanisms have not been well studied. In this study, we analyzed the distributions and phenotypic characteristics of three circulating monocyte subsets--CD14^＋＋CD16^-, CD14^＋＋CD16^＋ and CD14^＋＋mCD16^——in chronic HCV-infected patients, HCV spontaneous resolvers and healthy controls, and we evaluated the possible link between HCV viremia and disease progression. Our results indicated that the frequency of the CD 14^＋＋CD 16^＋ monocyte subset was decreased, and negatively correlated with HCV RNA and core antigen levels during chronic HCV infection. PD-L1 expression and the PD-L1/CD86 ratio in CD14^＋＋CD16^＋ monocytes were higher during chronic HCV infection than in spontaneous HCV resolvers and healthy controls. The PD-L1/CD86 ratio positively correlated with HCV viral load and core antigen levels. Finally, PD-L1 was significantly increased, while cytokine secretions were dramatically decreased upon Toll-like receptor （TLR） ligand binding and HCV JFH-lstimulation. These findings indicates the compromised immune status of the CD14^＋＋CD16^＋ monocytes during chronic HCV infection and provides new insights into the specific role of the CD14^＋＋CD16^＋ monocytes and their significance in chronic HCV infection.

A winning war against hepatitis B virus infection in China

Hepatitis B virus （HBV） infection is a worldwide health problem. According to WHO estimation,about 2 billion people globally have been infected with HBV and 350 million of them have chronic HBV infection. Like in other Asia-Pacific countries where HBV infection is highly endemic, chronic HBV infection causes enormous economic and social burdens in China. The national seroepidemiological survey on viral hepatitis conducted in 1992 showed that the prevalence of hepatitis B surface antigen （HBsAg） carriage status was 9.75% in general population. It is estimated that each year 300 000 deaths in China are caused by chronic hepatitis B related conditions including liver cirrhosis and hepatocellular carcinoma （HCC）. According to the 2007 Chinese Health Statistical Digest published by the Ministry of Health, in 2006 the incidence of viral hepatitis is the highest among the 35 infectious diseases that require compulsory report since 1978.