Hepatitis: Wang J

Tsukamoto H, She H, Hazra S, Cheng J, Wang J. · USC-UCLA Research Center of ALPD and Cirrhosis, Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. · J Gastroenterol Hepatol. · Pubmed #18336651 No free full text.

Abstract: Alcoholic and non-alcoholic steatohepatitis (ASH and NASH) constitute two major types of chronic liver disease with worldwide prevalence and are histologically indistinguishable with shared pathogenetic mechanisms. More importantly, they have synergistic interactions for liver pathology. Comparative studies on ASH and NASH have been hampered by the use of different animal models with confounding variables, particularly those with extreme genetic, toxic, and malnutrition etiologies. The mouse intragastric model circumvents these problems and reproduces the natural course and etiological background of ASH and NASH. Further, our recent work reproduces a profound synergism between the two in the model. Intracellular accumulation of neural lipids is a hallmark biochemical feature of ASH and NASH. Although impaired lipid oxidation and export may contribute to this pathological change, enhanced lipogenic regulation is frequently encountered, as characterized by induction of lipogenic or adipogenic transcription factors (peroxisome proliferator-activated receptor [PPAR gamma], liver X receptor alpha[LXR alpha], sterol-regulatory element-binding protein-1c [SREBP-1c]). In contrast, we have recently defined transdifferentiation of hepatic stellate cells (HSC), a pivotal event in liver fibrogenesis, as an 'antilipogenic' or 'anti-adipogenic' phenomenon. Thus, there is an apparent paradox between hepatocytes and HSC in steatohepatitis in terms of the outcome of lipogenic regulation. Our recent work suggests that defective insulin signaling in activated HSC may be responsible for this paradox. Further, activated Wnt signaling is implicated in 'anti-adipogenic' stellate cell transdifferentiation in liver fibrogenesis.

Batey RG, Wang J. · Division of Medicine, John Hunter Hospital, Hunter Regional Mail Centre, Newcastle 2310, NSW Australia. · Front Biosci. · Pubmed #12086911 No free full text.

Abstract: The development of alcohol-induced liver injury is, in part, a consequence of the immunological/inflammatory response that alcohol stimulates. The abnormalities of immune function in heavy drinkers have been documented well. Cytokines, especially TNF alpha, produced from macrophages/Kupffer cells, play a role in the induction of liver cell necrosis and apoptosis. TNF alpha can cause liver cell apoptosis through the TNF alpha receptor or Fas/CD95 which is expressed by liver cells. Furthermore, chronic ethanol consumption may damage the liver by inhibiting the hepatotrophic and hepatoprotective actions of TNF alpha and other cytokines. There exists an intrinsic lymphocyte population in the normal liver. Intrahepatic T lymphocytes consist of a heterogeneous population of cells that has many and varied functional characteristics in addition to classical T cell activity. The population of intrahepatic T lymphocytes may arise via a thymus-independent pathway. Our recent work has demonstrated the role of liver-associated T lymphocytes in the pathogenesis of alcohol related liver injury initiated by a variety of stimuli such as endotoxin (lipopolysaccharide, LPS) or concanavalin A (Con A). Our studies have, for the first time, suggested that alcohol consumption alone does not lead to the development of marked liver necrosis (at least in the rat), but rather that a second insult is required for this to occur. Liver-associated T lymphocytes in rats spontaneously secrete interleukin-1 alpha, interleukin-6 and TNF alpha in vitro culture. There is a significant decline in the amounts of interleukin-1 alpha and TNF alpha secreted in ethanol-consuming rats compared with non-ethanol consuming rats. The numbers of T cells, NK cells and Kupffer cells in liver perfusates remains stable over a prolonged period of ethanol consumption. However, following Con A injection, there was an inappropriate increase in the amounts of interleukin-6 and TNF alpha secreted in in vitro culture of liver-associated T lymphocytes and a significant increase in the percentage of CD4+ T cells and CD25+ T cells in liver perfusates compared with non-ethanol consuming rats. It suggested that liver-associated T lymphocytes are involved in the inflammatory process associated with alcohol related liver injury through increased cytokine secretion (TNF alpha).

Cheng AL, Kang YK, Chen Z, Tsao CJ, Qin S, Kim JS, Luo R, Feng J, Ye S, Yang TS, Xu J, Sun Y, Liang H, Liu J, Wang J, Tak WY, Pan H, Burock K, Zou J, Voliotis D, Guan Z. · National Taiwan University Hospital, Taipei, Taiwan. · Lancet Oncol. · Pubmed #19095497 No free full text.

Abstract: BACKGROUND: Most cases of hepatocellular carcinoma occur in the Asia-Pacific region, where chronic hepatitis B infection is an important aetiological factor. Assessing the efficacy and safety of new therapeutic options in an Asia-Pacific population is thus important. We did a multinational phase III, randomised, double-blind, placebo-controlled trial to assess the efficacy and safety of sorafenib in patients from the Asia-Pacific region with advanced (unresectable or metastatic) hepatocellular carcinoma. METHODS: Between Sept 20, 2005, and Jan 31, 2007, patients with hepatocellular carcinoma who had not received previous systemic therapy and had Child-Pugh liver function class A, were randomly assigned to receive either oral sorafenib (400 mg) or placebo twice daily in 6-week cycles, with efficacy measured at the end of each 6-week period. Eligible patients were stratified by the presence or absence of macroscopic vascular invasion or extrahepatic spread (or both), Eastern Cooperative Oncology Group performance status, and geographical region. Randomisation was done centrally and in a 2:1 ratio by means of an interactive voice-response system. There was no predefined primary endpoint; overall survival, time to progression (TTP), time to symptomatic progression (TTSP), disease control rate (DCR), and safety were assessed. Efficacy analyses were done by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00492752. FINDINGS: 271 patients from 23 centres in China, South Korea, and Taiwan were enrolled in the study. Of these, 226 patients were randomly assigned to the experimental group (n=150) or to the placebo group (n=76). Median overall survival was 6.5 months (95% CI 5.56-7.56) in patients treated with sorafenib, compared with 4.2 months (3.75-5.46) in those who received placebo (hazard ratio [HR] 0.68 [95% CI 0.50-0.93]; p=0.014). Median TTP was 2.8 months (2.63-3.58) in the sorafenib group compared with 1.4 months (1.35-1.55) in the placebo group (HR 0.57 [0.42-0.79]; p=0.0005). The most frequently reported grade 3/4 drug-related adverse events in the 149 assessable patients treated with sorafenib were hand-foot skin reaction (HFSR; 16 patients [10.7%]), diarrhoea (nine patients [6.0%]), and fatigue (five patients [3.4%]). The most common adverse events resulting in dose reductions were HFSR (17 patients [11.4%]) and diarrhoea (11 patients [7.4%]); these adverse events rarely led to discontinuation. INTERPRETATION: Sorafenib is effective for the treatment of advanced hepatocellular carcinoma in patients from the Asia-Pacific region, and is well tolerated. Taken together with data from the Sorafenib Hepatocellular Carcinoma Assessment Randomised Protocol (SHARP) trial, sorafenib seems to be an appropriate option for the treatment of advanced hepatocellular carcinoma.

Yan L, Han Y, Wang J, Liu J, Hong L, Fan D. · Department of Gastroenterology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi Province, China. · Am J Hematol. · Pubmed #17724706 No free full text.

Abstract: Peripheral blood monocytes (PBMCs) have the potential to differentiate into various progenitor cells. Here we have investigated the differentiation potential of PBMCs derived from patients with HBV related decompensated liver cirrhosis into hepatocyte-like cells. In our clinical trial, the PBMCs from 2 patients were mobilized by the recombinant human granulocyte colony stimulating factor, followed by leukapheresis and transplantation of PBMCs. PBMCs, induced by recombinant human hepatocyte growth factors, were identified by the expression of hepatocyte markers and specific biological functions with biochemical assays in vitro. Patients showed a lasting clinical amelioration for more than one year after transplantation, and hepatocyte-like cells were identified by expressing liver specific genes, synthesizing albumin, urea, aspirate transaminase, and glycogen, which were all similar to the human normal hepatic cell line QZG. Our results clearly demonstrated that mobilized PBMCs from patients with HBV related decompensated liver cirrhosis could differentiate into functional hepatocyte-like cells, indicating the possibility of autologous cell transplantation for treating patients with HBV related decompensated liver cirrhosis.

Hong L, Zhao Y, Han Y, Guo W, Wang J, Li X, Han Y, Fan D. · State Key Laboratory of Cancer Biology and Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China. · Helicobacter. · Pubmed #17669102 No free full text.

Abstract: Helicobacter pylori infection might be associated with vascular diseases, such as primary Raynaud phenomenon and coronary heart diseases. The possible mechanism might be due to H. pylori antigens causing intermittent vasospasm of arterioles, which also played roles in the development of liver cirrhosis. Migraine, a functional vascular disease, was observed in many patients with cirrhosis in the clinic. This study aimed to assess the effects of H. pylori eradication on migraine symptoms in patients with hepatitis-B-virus-related cirrhosis. The results clearly showed that the intensity, duration, and frequency of attacks of migraine were significantly reduced in all the patients in whom H. pylori has been eradicated. Thus, the study pushed further insight into the mechanisms of migraine pathogenesis.

Wang J, Xiang GJ, Liu BX. · Department of Aetiology and Immunology, Anhui University of Science and Technology, Huainan 232001, Anhui Province, China. · World J Gastroenterol. · Pubmed #12679925 links to  free full text

Abstract: AIM: To study the level of membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMC) and the therapeutic efficacy of alpha 2b interferon on the treatment of HCV-RNA in PBMC of patients with chronic hepatitis C and to compare the negative rates of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum. METHODS: Before and after treatment of alpha 2b interferon, the level of mIL-2R of patients with chronic hepatitis C was detected by biotin-streptavidin (BSA). The therapeutic group (26 cases) was treated with alpha 2b interferon (3 MU/d) and control therapeutic group (22 cases) was treated with routine drugs (VitC, aspartic acid). The total course of treatment with alpha 2b interferon and routine drug was six months and per course of the treatment was three months. The levels of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum were detected before and after a course of the treatment. RESULTS: Before and after treatment of alpha 2b interferon and routine drugs, the levels of mIL-2R in silence stage were (3.44+/-0.77) % and (2.95+/-0.72) %, the levels of mIL-2R in inducement stage were (33.62+/-3.95) % and (30.04+/-3.73) %. There was a significant difference between two groups (P<0.01-P<0.05). After treatment of alpha 2b interferon with 3 MU/d for two courses of the treatment, the total negative rates of HCV-RNA in the PBMC and HCV-RNA, anti-HCV in serum were 42.31 % (11/26), 57.69 % (15/26), 65.38 %(17/26) respectively. After the treatment of routine drug, the negative rates of HCV-RNA in PBMC and HCV-RNA, anti-HCV in serum were 13.64 % (3/22), 22.73 % (5/22), 27.27 % (6/22) respectively. There was high significant difference in the group treated with alpha 2b interferon and the group treated with routine drugs (P<0.01-P<0.05). CONCLUSION: The mIL-2R can be induced by alpha 2b interferon during the treatment. The alpha 2b interferon has a definite effect on the treatment of HCV-RNA in PBMC. The curative effect of alpha 2b interferon is better than that of the routine drugs.

Wang J, Zhu Q, Zhang T, Yu H. · Department of Infectious Disease, Children's Hospital, Fudan University, Shanghai 200032, China. · Chin Med J (Engl). · Pubmed #12622932 links to  free full text

Abstract: OBJECTIVE: To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte-macrophage colony-stimulating factor (GM-CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac). METHODS: A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 micro g of GM-CSF plus 10 micro g of rHBvac injected intramuscularly at the same location (GM-CSF group, 14 children) or 200 IU HBIG and 10 micro g rHBvac in different muscles (HBIG group, 13 children) on a monthly four-dose schedule. HBV-DNA quantification and other HBV serological markers were tested before and after the four-dose therapy. RESULTS: Twelve children in each group completed the study. Of them, 3 children in the GM-CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment. Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM-CSF group and 10 in the HBIG group. One from each group had an HBeAg/anti-HBe seroconversion after the treatment. The quantity of HBV-DNA was significantly lower after the treatment (P = 0.023) in GM-CSF group, but was not significantly reduced in HBIG group. No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group. CONCLUSION: Combined GM-CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis.

Du F, Liu QL, Fu QP, Sun L, Ao R, Guan XJ, Liu Y, Wang J, He H, Tong WB, Qin ZY, Fan WJ, Li J, He JL, Fang G. · Sichuan Province Center for Disease Control and Prevention, Chengdu 610041, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #19565873 No free full text.

Abstract: OBJECTIVE: To understand the changes of hepatitis B infection rates, before and after the hepatitis B vaccine was included into EPI, and to evaluate the effect of immunization which would lead to the development of a more appropriate hepatitis B control strategy. METHODS: Seroepidemiologic method, with multi-section random sampling method were chosen. 14 sites from 8 counties were involved. 2-4 ml of the vein blood was drawn from all the individuals engaged in the study including 3806 samples. HBsAg, anti-HBs, anti-HBc of the samples were tested with ELISA. RESULTS: Standardized positive rates of HBsAg and HBsAb were found as 7.05% and 29.77% respectively with the overall infection rate of HBV as 40.30%. The hepatitis B vaccine coverage of the children under 15 years was 70.73% and the positive rates for both HBsAg and anti-HBs were 2.62% and 56.68%, respectively. The coverage of hepatitis B vaccine among children under 3 years was 83.44% and the positive rates of both HBsAg and anti-HBs were 1.47% and 67.69% respectively, hepatitis B vaccine coverage of children under 3 years was 85.77%, with positive rates of HBsAg and anti-HBs as 1.78% and 75.44% respectively. CONCLUSION: Results from our study revealed that since the introduction of hepatitis B vaccination, the prevalence rates of HBsAg and HBV infection had an obvious decline, especially in children aged under 15 years of old, suggesting that some changes had occurred in the epidemic characteristics of hepatitis B in Sichuan.

Xu Y, Hu Y, Shi B, Zhang X, Wang J, Zhang Z, Shen F, Zhang Q, Sun S, Yuan Z. · Fudan University, China. · Mol Immunol. · Pubmed #19501403 No free full text.

Abstract: Plasmacytoid dendritic cells (pDCs), the professional producers of type I interferons (IFN-alpha/beta), play a pivotal role in innate and adaptive immune responses against viral infections. Although functional impairment of circulating pDCs in chronic hepatitis B (CHB) patients has been reported previously, the mechanism responsible for these defects remains unclear. We hypothesize that HBsAg circulating in high amounts during HBV infection may interact with pDC and contribute to pDC dysfunction. In support of this hypothesis we show that pDCs treated with HBsAg secreted much less IFN-alpha than control pDCs. Furthermore, suppression is specific for TLR9, with no effects upon TLR7-mediated IFN-alpha secretion. HBsAg inhibited TLR9-mediated IRF-7 expression and nuclear translocation, which are important for induction of IFN-alpha gene transcription. HBsAg upregulated the SOCS-1 expression and bound to BDCA-2 receptors on the plasma membrane of pDCs, resulting in the inhibition of the IFN-alpha production. In conclusion, the above data suggested that HBsAg may directly interfere with the function of pDC through HBsAg-mediated upregulation of SOCS-1 expression and BDCA-2 ligation, which could partially explain how HBV evades the immune system to establish a persistent infection.

Sheng XC, Pyun HJ, Chaudhary K, Wang J, Doerffler E, Fleury M, McMurtrie D, Chen X, Delaney WE, Kim CU. · Gilead Sciences, Inc, 333 Lakeside Drive, Foster City, CA 94404, USA. · Bioorg Med Chem Lett. · Pubmed #19477126 No free full text.

Abstract: A novel class of phosphonate derivatives was designed to mimic the interaction of product-like carboxylate based inhibitors of HCV NS3 protease. A phosphonic acid (compound 2) was demonstrated to be a potent HCV NS3 protease inhibitor, and a potential candidate for treating HCV infection. The syntheses and preliminary biological evaluation of this phosphonate class of inhibitor are described.

Saltanat H, Li H, Xu Y, Wang J, Liu F, Geng XH. · The Clinical Laboratory of the Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830028, China. · Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. · Pubmed #19426601 No free full text.

Abstract: AIM: To investigate the influences of camel milk on the immune response of the chronic hepatitis B patients and its possible mechanism. METHODS: After drinking camel milk for one year, 44 chronic hepatitis B patients were observed and the HBV-DNA, hepatitis B virus markers, ALT, IL-4 and INF-gamma levels in serum were detected. 60 chronic hepatitis B patients without any interventions for 1 year were taken as control. RESULTS: The level of Th1-type cytokine IFN-gamma in camel milk drinking group was significantly higher than that in the non-drinking camel milk group (P<0.05), however, the level of Th2-type cytokines IL-4 in camel milk drinking group was significantly lower than that in the non-drinking camel milk group (P<0.01). Both IFN-gamma and IL-4 levels in camel milk drinking group were near to those in the normal control group. The HBV-DNA negative rate of the camel milk drinking group (90.91%) was significantly higher than that of the non-drinking group (3.23%) (P<0.01). The HBsAg negative rates of the camel milk drinking group (54.55%) was also higher than that of the non-drinking group (1.61%)(P<0.01).The ALT level of 44 cases in the camel milk drinking group (100%)and 7 cases in the non-drinking group(11.29%) turned back to the normal level, there was a significant difference between the two group (P<0.01). CONCLUSION: Camel milk regulates the expression of Th1/Th2-type cytokines, and corrects the imbalance of Th1/Th2 cytokine network, which could strengthen the cellular immune response, inhibit the replication of virus DNA, and promote the recovery of the chronic hepatitis B patients.

Wang J, Jiang D, Zhang H, Lv S, Rao H, Fei R, Wei L. · Peking University Hepatology Institute, Peking University People's Hospital, Beijing, China. · Proteomics. · Pubmed #19242931 No free full text.

Abstract: Hepatitis B virus (HBV) infection is a worldwide health problem and may develop to liver fibrosis, cirrhosis, and even hepatocellular carcinoma. To investigate the global proteome responses of liver-derived cells to HBV infection and IFNalpha treatment, 2-DE and MS-based analysis were performed to compare the proteome changes between HBV stably transfected cell line HepG2.2.15 and its parental cell line HepG2, as well as HepG2.2.15 before and after IFNalpha treatment (5000 IU/mL for 72 h). Compared to HepG2, 12 of 18 down-regulated and 27 of 32 up-regulated proteins were identified in HepG2.2.15. After IFNalpha treatment, 6 of 7 down-regulated and 11 of 14 up-regulated proteins were identified. Differentially expressed proteins caused by HBV infection were involved with cytoskeletal matrix, heat shock stress, kinases/signal transduction, protease/proteasome components, etc. Prohibitin showed a dose-dependent up-regulation during IFNalpha treatment and might play a potent role in anti-HBV activities of IFNalpha by enhancing the crossbinding p53 expression to achieve the apoptosis of HBV infected liver cells. Down-regulation of interferon-stimulated gene 15 (ISG15) in HepG2.2.15 and recovery by IFNalpha suggested its relationship with IFNalpha's anti-HBV effect.

Robinson RT, Wang J, Cripps JG, Milks MW, English KA, Pearson TA, Gorham JD. · Department of Microbiology and Immunology, Dartmouth Medical School, Lebanon, NH 03756, USA. · J Immunol. · Pubmed #19234226 No free full text.

Abstract: Fulminant inflammation in the liver is often accompanied by the accumulation of IFN-gamma-producing T cells. The BALB/c-Tgfb1(-/-) mouse exhibits extensive, spontaneously developing necroinflammation in the liver, accompanied by the accumulation of IFN-gamma-producing CD4(+) and CD8(+) T cells. Liver damage depends on the presence of an intact Ifng gene. We determined the relevant cellular source(s) of IFN-gamma. In Tgfb1(-/-) liver, CD4(+) T cells were more numerous than CD8(+) T cells and NK cells, and produced more IFN-gamma. Depletion of CD4(+) T cells eliminated both the elevation in plasma IFN-gamma and aspartate aminotransferase, whereas depletion of CD8(+) T cells did not. Rag1(-/-)Tgfb1(-/-) mice exhibited neither IFN-gamma elevation nor tissue damage, indicating that NK cells are not sufficient. IFN-gamma was required for strong overexpression of class II genes but not for CD4(+) T cell activation, oligoclonal expansion, or accumulation in the liver. The T cell inhibitory molecule PD-L1 was strongly expressed in Tgfb1(-/-) livers, ruling out a lack of PD-L1 expression as an explanation for aberrant liver T cell activation. Finally, whereas Tgfb1(-/-) CD4(+) T cells overexpressed Fas ligand, hepatocellular damage was observed in Fas(lpr/lpr)Tgfb1(-/-) mice, indicating that liver pathology is Fas independent. We conclude that liver damage in this model of fulminant autoimmune hepatitis is driven by CD4(+) T cell production of IFN-gamma, is independent of both CD8(+) T cells and the Fas ligand/Fas pathway, and is not explained by a lack of PD-L1 expression.

Jin H, Wang J, Yan L, Nie JJ, Li J, Zhuang H. · Department of Microbiology, Peking University Health Science Center, Beijing 100191, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #19173971 No free full text.

Abstract: OBJECTIVE: To establish a hepatitis B virus (HBV) nested PCR (nPCR) for detection of genotypes A-D and subgenotypes B1, B2, C1 and C2. METHODS: The entire HBV nucleotide sequences of genotypes A-H retrieved from GenBank were compared and analyzed by DNAStar software. The PCR primers were designed by Primer Premier 5.0 software, and the nPCR for genotyping HBV/A-D as well as subgenotyping B1, B2, C1 and C2 were established. There were 3 steps in the process:step 1 for genotypes B, D and subgenotypes C1, C2 with the amplification of Mix A; step 2 for genotype A with the amplification of Mix B; step 3 for subgenotypes B1 and B2 with the amplification of Mix C in the second-round PCR, based on the first-round amplification procedure. A total of 68 serum samples from patients with chronic HBV infection were detected by nPCR. 15 of 68 sera were selected randomly and their PCR products were directly sequenced to confirm the accuracy of the method. RESULTS: Among 68 serum samples of patients with chronic HBV infection detected by the nPCR, 23.53% (16/68) were infected with B2, 11.76% (8/68) with C1, 48.53% (33/68) with C2, 1.47% (1/68) with D, 11.76% (8/68) with B2C2 mix strains, 1.47% (1/68) with C2D mix strains and 1.47% (1/68) with B2/C1/D mix strains. The sequencing analysis of the 15 serum samples had the same results as detected by nPCR. CONCLUSION: nPCR is a simple, rapid method and able to detect genotypes A-D and subgenotypes B1, B2, C1 and C2 subtypes of HBV with both high sensitivity and specificity.

Kim HJ, Sharon A, Bal C, Wang J, Allu M, Huang Z, Murray MG, Bassit L, Schinazi RF, Korba B, Chu CK. · The University of Georgia College of Pharmacy, Athens, Georgia 30602, USA. · J Med Chem. · Pubmed #19072694 No free full text.

Abstract: A series of 7-deazaneplanocin A (7-DNPA, 2) analogues were synthesized and evaluated for in vitro antiviral activity against HBV and HCV. The syntheses of target carbocyclic nucleosides were accomplished via a convergent procedure. 7-Substitutions were introduced by using 7-substituted-7-deaza heterocyclic base precursors (F, Cl, Br, and I) or via substitution reactions after the synthesis of the carbocyclic nucleosides. Among the synthesized compounds, 2, 13-15, 24, and 27 exhibited significant anti-HCV activity (EC(50) ranged from 1.8 to 20.1 microM) and compounds 2, 15, 22, and 24 demonstrated moderate to potent anti-HBV activity (EC(50) = 0.3-3.3 microM). In addition, compound 24 also showed activity against lamivudine- and adefovir-associated HBV mutants.

Liu K, Qian L, Wang J, Li W, Deng X, Chen X, Sun W, Wei H, Qian X, Jiang Y, He F. · State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China. · Mol Cell Proteomics. · Pubmed #18984579 No free full text.

Abstract: Hepatitis B virus (HBV) infection is a major health concern with more than two billion individuals currently infected worldwide. Despite the prevalence of infection, gaining a complete understanding of the molecular mechanisms of HBV infection has been difficult because HBV cannot infect common immortalized cell lines. HepG2.2.15, however, is a well established version of the HepG2 cell line that constitutively expresses HBV. Therefore, comparative proteomics analysis of HepG2.2.15 and HepG2 may provide valuable clues for understanding the HBV virus life cycle. In this study, two-dimensional blue native/SDS-PAGE was utilized to characterize different multiprotein complexes from whole cell lysates between HepG2.2.15 and HepG2. These results demonstrate that two unique protein complexes existed in HepG2.2.15 cells. When these complexes were excised from the gel and subjected to the second dimension separation and the proteins were sequenced by mass spectrometry, 20 non-redundant proteins were identified. Of these proteins, almost 20% corresponded to heat shock proteins, including HSP60, HSP70, and HSP90. Antibody-based supershift assays were used to verify the validity of the distinct protein complexes. Co-immunoprecipitation assays confirmed that HSP60, HSP70, and HSP90 proteins physically interacted in HepG2.2.15 but not HepG2 cells. We further demonstrated that down-regulation of HSP70 or HSP90 by small interfering RNA significantly inhibited HBV viral production but did not influence cellular proliferation or apoptosis. Consistent with these results, a significant reduction in HepG2.2.15 HBV secretion was observed when the HSP90 inhibitor 17-allylamino-17-demethoxygeldanamycin was used to treat HepG2.2.15 cells. Collectively these results suggest that the interaction of HSP90 with HSP70/HSP60 contributes to the HBV life cycle by forming a multichaperone machine that may constitute therapeutic targets for HBV-associated diseases.

Wei B, Wei Y, Zhang K, Wang J, Xu R, Zhan S, Lin G, Wang W, Liu M, Wang L, Zhang R, Li J. · National Center for Clinical Laboratories, Beijing Hospital, Beijing, People's Republic of China. · Biomed Pharmacother. · Pubmed #18823738 No free full text.

Abstract: RNA-based therapeutic strategies are used widely due to their highly specific mode of action. However, the major obstacle in any RNA-based therapy is cellular delivery and stability in the cells. The self-assembly of the MS2 bacteriophage capsids has been used to develop virus-like particles (VLPs) for drug delivery. In this study, we utilized the heterobifunctional crosslinker, sulfosuccinimidyl-4-(p-maleimidophenyl)-butyrate (sulfo-SMPB), to conjugate the human immunodeficiency virus-1 (HIV-1) Tat peptide and MS2 VLPs; the antisense RNA against the 5'-untranslated region (UTR) and the internal ribosome entry site (IRES) of the hepatitis C virus (HCV) was packaged into these particles by using a two-plasmid coexpression system. The MS2 VLPs conjugated with the Tat peptide were then transferred into Huh-7 cells containing an HCV reporter system. The packaged antisense RNA showed an inhibitory effect on the translation of HCV. This paper describes our initial results with this system using the Tat peptide.

Wang J, Liu DW, Kong FZ. · School of Public Health, Hebei Medical College, Shijiazhuang 050017, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18788539 No free full text.

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Wang J, Gao JW, Zhuang H, Li J, Wang J, Li YJ, Jin H. · Department of Microbiology, Peking University Health Science Center, Beijing 100083, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18785479 No free full text.

Abstract: OBJECTIVE: To investigate the distribution of sub-genotype B on hepatitis B virus (HBV) in patients with HBV chronic infection from 4 cities (Beijing, Shijiazhuang, Wenzhou and Shenzhen) of China. METHODS: A type-specific nested polymerase chain reaction(PCR) with multiplex pairs of primers was used for HBV genotyping,and Ba and Bj sub-genotypes were identified by a PCR-restriction fragment length polymorphism (PCR-RFLP) method. A total of 101 serum samples collected from patients with chronic HBV/B infection were detected. Among them, 18 were collected from Beijing, 22 from Shijiazhuang, 34 from Wenzhou and 27 from Shenzhen. Thirty from the 101 serum samples were randomly selected and analyzed by PCR product sequencing. RESULTS: All of the 101 serum samples were identified as sub-genotype Ba,and none of them was sub-genotype Bj. CONCLUSION: Sub-genotype Ba was a predominant strain of HBV/B in patients with chronic HBV infection from these 4 cities.

Wang J, Gao JW, Zhuang H, Li J, Wang J, Li YJ, Jin H. · Department of Microbiology, Peking University Health Science Center, Beijing 100083, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18785479 No free full text.

Abstract: OBJECTIVE: To investigate the distribution of sub-genotype B on hepatitis B virus (HBV) in patients with HBV chronic infection from 4 cities (Beijing, Shijiazhuang, Wenzhou and Shenzhen) of China. METHODS: A type-specific nested polymerase chain reaction(PCR) with multiplex pairs of primers was used for HBV genotyping,and Ba and Bj sub-genotypes were identified by a PCR-restriction fragment length polymorphism (PCR-RFLP) method. A total of 101 serum samples collected from patients with chronic HBV/B infection were detected. Among them, 18 were collected from Beijing, 22 from Shijiazhuang, 34 from Wenzhou and 27 from Shenzhen. Thirty from the 101 serum samples were randomly selected and analyzed by PCR product sequencing. RESULTS: All of the 101 serum samples were identified as sub-genotype Ba,and none of them was sub-genotype Bj. CONCLUSION: Sub-genotype Ba was a predominant strain of HBV/B in patients with chronic HBV infection from these 4 cities.

Wang J, Gao JW, Li J, Zhuang H, Wang J, Li YJ, Jin H. · Department of Microbiology, Peking University Health Science Center, Beijing 100083, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18686861 No free full text.

Abstract: OBJECTIVE: To establish a sensitive, specific, simple and practicable method for identifying the two sub-genotypes (Ba and Bj) of genotype B isolates of hepatitis B virus (HBV) (HBV/ B). METHODS: The entire nucleotide sequences of HBV/B and HBV/C were obtained from GenBank, compared and analyzed with DNAStar software. The specific primers for HBV/B (HB) and Ba (BA), Bj (BJ) were designed respectively. HB as HBV/B specific primer (sense) and HBAS-4V (designed by Japanese scientists Sugauchi et al) as a universal outer primer (antisense) were used in the first-round PCR. In the second-round PCR, HB was also used as sense primer while BA and BJ as inner primers (antisense) and they were added into a single tube for PCR reaction. The two sub-genotypes of HBV/B were identified according to the length of the PCR products. A total of 71 HBV DNA-positive serum samples were selected randomly from our laboratory, including 56 HBV/B samples identified by type-specified PCR method and 15 HBV/C samples identified by direct sequencing in preS and S Region (preS/S). All the 71 samples were detected with this semi-nested PCR method. A plasmid containing full length genomic DNA of HBV/Bj, was presented by Professor Kenji Abe as positive control of Bj. Then, the first-round PCR products of 15 HBV/B were randomly selected and sequenced directly, and their sequences were compared phylogenetically with the above known Ba and Bj sequences using Blast and DNAStar softwares to confirm the results of semi-nested PCR. RESULTS: 56 HBV/B samples were all identified as HBV/Ba by our semi-nested PCR method. 15 randomly selected PCR products were all sequenced as HBV/Ba. All of the 15 HBV/C samples were negative. CONCLUSION: A simple, rapid, sensitive and specific method for identifying sub-genotypes Ba and Bj was established with might be used for large-scale clinical and epidemiological studies.

Wang J, Gao JW, Li J, Zhuang H, Wang J, Li YJ, Jin H. · Department of Microbiology, Peking University Health Science Center, Beijing 100083, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18686861 No free full text.

Abstract: OBJECTIVE: To establish a sensitive, specific, simple and practicable method for identifying the two sub-genotypes (Ba and Bj) of genotype B isolates of hepatitis B virus (HBV) (HBV/ B). METHODS: The entire nucleotide sequences of HBV/B and HBV/C were obtained from GenBank, compared and analyzed with DNAStar software. The specific primers for HBV/B (HB) and Ba (BA), Bj (BJ) were designed respectively. HB as HBV/B specific primer (sense) and HBAS-4V (designed by Japanese scientists Sugauchi et al) as a universal outer primer (antisense) were used in the first-round PCR. In the second-round PCR, HB was also used as sense primer while BA and BJ as inner primers (antisense) and they were added into a single tube for PCR reaction. The two sub-genotypes of HBV/B were identified according to the length of the PCR products. A total of 71 HBV DNA-positive serum samples were selected randomly from our laboratory, including 56 HBV/B samples identified by type-specified PCR method and 15 HBV/C samples identified by direct sequencing in preS and S Region (preS/S). All the 71 samples were detected with this semi-nested PCR method. A plasmid containing full length genomic DNA of HBV/Bj, was presented by Professor Kenji Abe as positive control of Bj. Then, the first-round PCR products of 15 HBV/B were randomly selected and sequenced directly, and their sequences were compared phylogenetically with the above known Ba and Bj sequences using Blast and DNAStar softwares to confirm the results of semi-nested PCR. RESULTS: 56 HBV/B samples were all identified as HBV/Ba by our semi-nested PCR method. 15 randomly selected PCR products were all sequenced as HBV/Ba. All of the 15 HBV/C samples were negative. CONCLUSION: A simple, rapid, sensitive and specific method for identifying sub-genotypes Ba and Bj was established with might be used for large-scale clinical and epidemiological studies.

Fang SG, Shen H, Wang J, Tay FP, Liu DX. · Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673, Singapore. · Virology. · Pubmed #18678384 No free full text.

Abstract: Coronavirus 3C-like proteinase (3CLpro) plays important roles in viral life cycle through extensive processing of the polyproteins 1a and 1ab into 12 mature, non-structural proteins (nsp5-nsp16). Structural and biochemical studies have revealed that all confirmed 3CLpro cleavage sites have a conserved Gln residue at the P1 position, which is thought to be absolutely required for efficient cleavage. Recent studies on murine hepatitis virus (MHV) showed that processing of the 1a polyprotein at the position between nsp10-nsp11 is essential for viral replication. In this report, we investigated the requirement of processing at the equivalent position for replication of avian coronavirus infectious bronchitis virus (IBV), using an infectious cloning system. The results showed that mutation of the P1 Gln to Pro or deletion of the Gln residue in the nsp10-nsp11/12 site completely abolished the 3CLpro-mediated processing, but allowed production of infectious recombinant viruses with variable degrees of growth defect, suggesting that cleavage at the nsp10-nsp11/12 site of IBV is dispensable for viral replication in cultured cells. This study would pave a way for potential vaccine development by generation of attenuated IBV from field isolates through manipulation of the nsp10-nsp11/12 cleavage site. Similar approaches would be also applicable to other human and animal coronaviruses.

Yang J, Zhou L, Wang J, Wang G, Davey AK. · Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing, China. · Planta Med. · Pubmed #18671195 No free full text.

Abstract: The major component of liquorice root extract, glycyrrhizinate (GZ), has been formulated as an injection for the treatment of hepatitis. If given orally, GZ has poor bioavailability and is catalysed to glycyrrhetinic acid (GA) by intestinal bacteria. GA is subsequently responsible for significant side effects. This study was conducted to clarify the relationship between GZ and GA absorption and bioavailability. GZ and GA absorption were investigated using the in situ single pass isolated perfused intestine (IPI). Hepatic disposition was investigated using isolated perfused liver (IPL) and in vivo biliary excretion models. The apparent permeability and absorption rate constants in the IPI for GZ were 0.36 +/- 0.31 cm/min and 0.35 +/- 0.33 min (-1), while those for GA were 5.73 +/- 0.11 cm/min and 1.53 +/- 0.05 min (-1), respectively. The hepatic extraction ratios of unbound GZ and GA in the IPL were 0.22 +/- 0.01 and 0.44 +/- 0.15, respectively. Seven hours after intra-portal venous injection in vivo, the cumulative biliary excretion ratio for GZ was 96 %. There was negligible biliary excretion of unchanged GA during the same period. It was apparent in all models used that in the absence of intestinal bacteria GZ was not metabolised to GA, or vice versa. Hence, GZ can be absorbed unchanged from the intestine provided it has sufficient time and is protected from intestinal bacteria. This opens up the possibility that the use of pharmaceutical carrier systems or similar formulation approaches may allow effective oral administration of therapeutic levels of GZ without the side effects associated with GA.

Wang J, Li Q, Sun Y, Zheng H, Cui Y, Li H, Zhou H, Hao X. · Department of Hepatobiliary Surgery, Cancer Hospital of Tianjin Medical University, Huanhu Western Road, Hexi District, Tianjin 300060, PR China. · Surg Oncol. · Pubmed #18640032 No free full text.

Abstract: AIMS: To clarify the incidence of multicentric occurrence (MO) and intrahepatic metastasis (IM) for hepatocellular carcinoma (HCC) related to hepatitis B virus (HBV) in China and to identify the differences between them. PATIENTS AND METHODS: Histopathologic features of multiple tumors in 82 cases with HCC were analyzed. The two groups, the origin was determinable as of multicentric occurrence or as of intrahepatic metastasis, were analyzed for their survival rate, disease-free survival and clinicopathologic differences. RESULTS: According to histological findings, 19.5% and 69.5% patients were considered to be MO and IM, respectively. In total 73 cases from the histopathological method were selected and divided into group MO (16 cases) and the group IM (57 cases). Analysis of stepwise regression identified that: Child's stage, cholinesterase (host factors), tumor size, histological grade and positive portal vein invasion (tumor factors) were the most important discriminating factors between MO and IM (p<0.05). As for their prognosis, Kaplan-Meier and Log rank test showed the survival rate in group MO was significantly better than that in the group IM (p=0.003). No statistical significance was found between the disease-free survival in group MO and that in group IM (p=0.141). The analysis of Cox's proportional hazards model showed that tumor type (MO or IM) and Child's stage were the important prognostic factors (p=0.002 and 0.014, respectively). CONCLUSIONS: The incidence of MO in patients with multiple HCCs related to HBV is only about 20%, which is lower than that of Japan. Child's stage, cholinesterase (host factors), tumor size, histological grade and positive portal vein invasion (tumor factors) are the most important discriminating factors between MO and IM. The prognosis of patients with MO compared to IM is significantly better and tumor type (MO or IM) and Child's stage are important prognostic factors.