Hepatitis: Chen T

Chen T, Jia H, Li J, Chen X, Zhou H, Tian H. · Department of Endocrinology and Metabolism, West China Hospital of Sichuan University, Chengdu, China. · Transpl Int. · Pubmed #19207185 No free full text.

Abstract: New onset diabetes mellitus (NODM) postliver transplantation (LT) is very common and may negatively affect patient and graft survival, but its causative mechanism is still unclear. This study was to analyze the connection between Hepatitis C virus (HCV) infection and NODM after LT by systematically reviewing published medical literature. We electronically searched databases of MEDLINE, EMBASE and the Cochrane Library from January 1980 to January 2008. Only retrospective studies could be identified. Seven of them were subjected to the meta-analysis. Analysis was performed by using revman 4.2 software. We found that HCV increased the prevalence of NODM [OR 2.46; 95%CI (1.44, 4.19)]. Then, we further analyzed the association between HCV and persistent-NODM (P-NODM) after LT. The result showed that prevalence of P-NODM was higher in HCV-positive group than in HCV-negative group with marginally statistical significance [OR = 1.39; 95%CI (1.06, 1.83)]. The present meta-analysis based on retrospective studies suggested a significant relationship between HCV and NODM after LT, and it seems that HCV infection might also increase the prevalence of P-NODM. Multicenter, large sized prospective studies are still needed to further confirm these results.

McClain C, Barve S, Joshi-Barve S, Song Z, Deaciuc I, Chen T, Hill D. · Department of Internal Medicine, Department of Pharmacology and Toxicology, University of Louisville Medical Center, Louisville, KY 40202, USA. · Alcohol Clin Exp Res. · Pubmed #16344606 No free full text.

Abstract: Alcoholic liver disease (ALD) remains an important complication and cause of morbidity and mortality from alcohol abuse. Major developments in our understanding of the mechanisms of ALD over the past decade are now being translated into new forms of therapy for this disease process which currently has no FDA approved treatment. Cytokines are low molecular weight mediators of cellular communication, and the pro-inflammatory cytokine tumor necrosis factor (TNF) has been shown to play a pivotal role in the development of experimental ALD. Similarly, TNF levels are elevated in the serum of alcoholic hepatitis patients. Abnormal methionine metabolism is well documented in patients with ALD, with patients having elevated serum methionine levels, but low S-adenosylmethionine levels in the liver. On the other hand, S-adenosylhomocysteine and homocysteine levels are elevated in ALD. Recent studies have documented potential interactions between homocysteine and S-adenosylhomocysteine with TNF in the development of ALD. Altered proteasome function also is now well documented in ALD, and decreased proteasome function can cause hepatocyte apoptosis. Recently it has been shown that decreased proteasome function can also act synergistically to enhance TNF hepatotoxicity. Hepatocytes dying of proteasome dysfunction release pro-inflammatory cytokines such as Interleukin-8 to cause sustained inflammation. This article reviews the interactions of cytokines, altered methionine metabolism, and proteasome dysfunction in the development of ALD.

McClain CJ, Mokshagundam SP, Barve SS, Song Z, Hill DB, Chen T, Deaciuc I. · Department of Internal Medicine, University of Louisville Medical Center, 530 South Jackson Street, ACB 3rd Floor, Louisville, KY 40292, USA. · Alcohol. · Pubmed #15670668 No free full text.

Abstract: In 1980, the term non-alcoholic steatohepatitis was coined to describe a new syndrome occurring in patients who usually were obese (often diabetic) females who had a liver biopsy picture consistent with alcoholic hepatitis, but who denied alcohol use. The causes of this syndrome were unknown, and there was no defined therapy. More than two decades later, this clinical syndrome is only somewhat better understood, and still there is no Food and Drug Administration-approved or even generally accepted drug therapy. Patients with primary non-alcoholic steatohepatitis typically have the insulin resistance syndrome (synonymous with the metabolic syndrome, syndrome X, and so forth), which is characterized by obesity, diabetes, hyperlipidemia, hypertension, and, in some instances, other metabolic abnormalities such as polycystic ovary disease. Secondary non-alcoholic steatohepatitis may be caused by drugs such as tamoxifen, certain industrial toxins, rapid weight loss, and so forth. The cause of non-alcoholic steatohepatitis remains elusive, but most investigators agree that a baseline of steatosis requires a second hit capable of inducing inflammation, fibrosis, or necrosis for non-alcoholic steatohepatitis to develop. Our research group has focused its efforts on the interactions of nutritional abnormalities, cytokines, oxidative stress with lipid peroxidation, and mitochondrial dysfunction in the induction of steatohepatitis, both alcoholic and non-alcoholic in origin. Research findings from other laboratories also support the role of increased cytokine activity, oxidative stress, and mitochondrial dysfunction in the pathogenesis of non-alcoholic steatohepatitis. The objectives of this article are to review the (1) definition and clinical features of non-alcoholic steatohepatitis, (2) potential mechanisms of non-alcoholic steatohepatitis, and (3) potential therapeutic interventions in non-alcoholic steatohepatitis.

Chen T, Chen WH. · Department of Basic Medicine, Yunnan College of Traditional Chinese Medicine, Kunming, Yunnan Province 650200, China. · Zhong Xi Yi Jie He Xue Bao. · Pubmed #15339592 links to  free full text

Abstract: This paper summarized the effects of the treatment of chronic hepatitis B (CHB) by lamivudine combined with traditional Chinese medicine or western medicine in the past few years. Combined treatment of lamivudine and the other medicine had higher efficiency than lamvudine alone in the treatment of CHB. The Combined treatment should be a tendency of the treatment for chronic hepatitis B. The focal point is that the traditional Chinese recipe produces a multitarget effect in the treatment of CHB. The combined treatment of lamivudine and traditional Chinese recipe can reduce the treatment course and the recurrence rate. Combined treatment of traditional Chinese medicine and western medicine has notable superiority and broad developing prospect in the treatment of CHB.

Chen T, Zamora R, Zuckerbraun B, Billiar TR. · Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA. · Curr Mol Med. · Pubmed #14527083 No free full text.

Abstract: The complex role of nitric oxide (NO) in the liver can be explained by its patterns of regulation and unique biochemical properties. With a broad range of direct and indirect molecular targets, NO acts as an inhibitor or agonist of cell signaling events. In the liver, constitutively generated NO maintains the hepatic microcirculation and endothelial integrity, while inducible NO synthase (iNOS)-governed NO production can be either beneficial or detrimental. For instance, NO potentiates the hepatic oxidative injury in warm ischemia/reperfusion, while iNOS expression protects against hepatic apoptotic cell death seen in models of sepsis and hepatitis. Anti-apoptotic actions are either cyclic nucleotide dependent or independent, including the expression of heat shock proteins, prevention of mitochondrial dysfunction, and inhibition of caspase activity by S-nitrosation. Whether NO protects or injures is probably determined by the type of insult, the abundance of reactive oxygen species (ROS), the source and amount of NO production and the cellular redox status of liver. Through the use of pharmacological NO donors or NOS gene transfer in conjunction with genetically altered knockout animals, the physiological and pathophysiological roles of NO in liver function can be explored in more detail. The purpose of this paper is to review the current understanding of the role of NO in liver injury.

McClain CJ, Hill DB, Song Z, Chawla R, Watson WH, Chen T, Barve S. · Department of Medicine, University of Louisville Medical Center, KY 40292, USA. · Alcohol. · Pubmed #12163148 No free full text.

Abstract: Hepatic deficiency of S-adenosylmethionine (AdoMet) is a critical acquired metabolic abnormality in alcoholic liver disease (ALD) and in many experimental models of hepatotoxicity. Subnormal AdoMet, elevated serum tumor necrosis factor (TNF), and endotoxemia (LPS) are hallmarks of ALD and experimental liver injury. AdoMet deficiency is attributed to its subnormal synthesis, but mechanisms for increased TNF are not known. AdoMet deficiency may affect the critical balance of proinflammatory (e.g., TNF) and antiinflammatory [e.g., interleukin (IL)-10] cytokines. Rats maintained on a choline-deficient diet with limited amounts of methionine (MCD diet) developed AdoMet deficiency. When challenged with LPS, rats fed MCD diet had significantly increased serum TNF levels and worse liver injury compared with findings for controls. Exogenous AdoMet attenuated liver injury and serum TNF levels. Results of in vitro studies with the use of RAW 264.7 cells demonstrated that exogenous AdoMet supplementation lowered LPS-induced TNF formation in a dose-dependent manner, and AdoMet deficiency enhanced TNF secretion and TNF gene expression. AdoMet also dose-dependently decreased LPS-stimulated TNF production from monocytes obtained from patients with alcoholic hepatitis. Finally, AdoMet supplementation stimulated production of the antiinflammatory cytokine IL-10. Interleukin-10 plays a critical role in the modulation of TNF production, and IL-10 may inhibit hepatic fibrosis. This article will review (1) the role of AdoMet in ALD/liver injury, (2) the role of TNF/proinflammatory cytokines in ALD, (3) potential roles of AdoMet in TNF/proinflammatory cytokine regulation in ALD, and (4) conclusions and future directions.

Sun Y, Qin Y, Gong FY, Wu XF, Hua ZC, Chen T, Xu Q. · State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 22 Han Kou Road, Jiangsu, Nanjing 210093, China. · Biochem Pharmacol. · Pubmed #19428326 No free full text.

Abstract: Selectively inducing apoptosis of activated T cells is essential for the clearance of pathogenic injurious cells and subsequent efficient resolution of inflammation. However, few chemicals have been reported to trigger apoptosis of activated T cells in the treatment of hepatitis without affecting quiescent T cells. In the present study, we found that fraxinellone, a small natural compound isolated from the root bark of Dictamnus dasycarpus, selectively facilitated apoptosis of concanavalin A (Con A)-activated CD4(+) T cells rather than those non-activated, by disrupting the mitochondrial transmembrane potential, decreasing the ratio of Bcl-2/Bax, and increasing cytochrome c release from the mitochondria to the cytosol. The enhancement in Fas expression and caspase-8 activity, truncation of Bid, and down-regulation of anti-apoptotic cellular FLICE-inhibitory protein expression by fraxinellone also suggested the participation of an extrinsic apoptosis pathway. Furthermore, fraxinellone significantly alleviated Con A-induced T-cell-dependent hepatitis in mice, which was closely associated with reduced serum transaminases, pro-inflammatory cytokines, and pathologic parameters. Consistent with the in vitro results, fraxinellone dramatically induced apoptosis of activated peripheral CD4(+) T cells in vivo, consequently resulting in less CD4(+) T-cell activation and infiltration to the liver. These results strongly suggest fraxinellone might be a potential leading compound useful in treating T-cell-mediated liver disorders in humans.

Zhou YY, Pi B, Liu XJ, Zhang R, Deng GH, Chen T, Wang HW, Han MF, Yan WM, Xi D, Huang JQ, Wang YM, Luo XP, Ning Q. · Department of Infectious Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #19032868 No free full text.

Abstract: 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.

Liu J, Li Y, Chen T, Yang Y, Wang K, He Y, Yang Q, Ye F, Jin Y, Qiu T, Lin S, Liu M, Zhao Y. · Department of Infectious Diseases, The First Affiliated Hospital of Xi'an JiaoTong University School of Medicine, 710061, Xi'an, Shaanxi Province, China. · Arch Virol. · Pubmed #18982245 No free full text.

Abstract: Using a nested-PCR genotyping system, we analyzed the viral genotypes in HBV-infected patients from HBV clustering infection families (CIFs), as compared with HBV infected patients without familial infection history. The patients in the CIF group showed significantly increased prevalence of genotype C infection, while genotype B was absent. Additionally, in the genotype C carriers younger than 50 years old, the prevalence of HCC and LC were significantly increased in CIF compared to the control. We found that HBeAg-positive HBV genotype C carriers are prone to develop end-stage liver diseases earlier than those infected by genotype B.

Zou Y, Chen T, Wang HW, Yan WM, Luo XP, Ning Q. · Department of Infectious Diseases, Institute of Infectious Diseases, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #18822205 No free full text.

Abstract: OBJECTIVES: To investigate the role of liver natural killer cells (NK cells) in murine hepatitis virus strain 3 (MHV-3) induced murine fulminant hepatitis. METHOD: Balb/cJ mice (6-8 weeks, female) were intraperitoneally injected with 100 PFU MHV-3. The numbers of NK cells in their livers, spleens, blood and bone marrow and the expression of CD69 on liver NK cells at 0, 24, 48 and 70 h after MHV-3 infection were analyzed by flow cytometry. The cytotoxic activity of liver NK cells was detected by a non-radioactive cytotoxicity assay. The levels of IFN gamma produced by hepatic NK cells were detected by intracellular cytokine staining. RESULT: Following MHV-3 infection, the proportion of liver NK cells in the mice increased remarkably and reached the peak (43.9%+/-2.3%) at 48 h, then kept a high proportion until the mice were sacrificed. The proportion of NK cells in the peripheral blood also significantly increased and reached the peak (18.0%+/-5.4%) at 48 h. However, there were few NK cells in the peripheral blood at 70 h after infection; the ratio was only 1.3%+/-0.6%. In the spleens and bone marrow, the proportions of NK cells were both significantly decreased from 0 h to 48 h and then slightly increased. The expression of CD69 on liver NK cells was highly up-regulated after the infection and the cytotoxic activity of hepatic NK cells at 48 h was also significantly enhanced. In addition, an increase in IFN gamma production by hepatic NK cells was observed at 48 h. CONCLUSION: After MHV-3 infection, NK cells were recruited to the liver quickly, probably from the spleen and bone marrow. Recruited NK cells remarkably express CD69, enhance cytotoxic activity and IFN gamma production, which correlate with the disease severity of fulminant viral hepatitis. Our results suggest that liver NK cells may play a pivotal role in the pathogenesis of fulminant viral hepatitis.

He XX, Chen T, Lin JS, Chang Y, Ye BX. · Institute of Liver Diseases, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. · Biochem Biophys Res Commun. · Pubmed #18284916 No free full text.

Abstract: Antiviral therapy of chronic hepatitis B remains a major clinical problem worldwide. Like lamivudine, nucleoside analogs have become the focus of investigation of anti-hepatitis B virus (anti-HBV) drugs. Here, beta-LPA is a novel 2,6-diaminopurine analog found to possess potent anti-HBV activity. In HepG2.2.15 cell line, beta-LPA had a 50% effective concentration (EC(50)) of 0.01 microM against HBV, as determined by analysis of secreted and intracellular episomal HBV DNA. Levels of HBV surface antigen (HBsAg) and e antigen (HBeAg) in drug-treated cultures revealed that beta-LPA had no significant inhibitory effects on HBsAg and HBeAg. beta-LPA didn't show any cytotoxicity up to 0.4 microM with a 50% cytotoxic concentration (CC(50)) of 50 microM. Furthermore, treatment with beta-LPA resulted in no apparent inhibitory effects on mitochondrial DNA content. Considering the potent inhibition of HBV DNA synthesis and no obvious toxicity of beta-LPA, this compound should be further explored for development as an anti-HBV drug.

Chen T, Chen M, Wang LP, Feng B, Li XH. · Department of Infectious Diseases, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China. · Zhonghua Gan Zang Bing Za Zhi. · Pubmed #16635313 No free full text.

This publication has no abstract.

Ye F, Yue Y, Li S, Chen T, Bai G, Liu M, Zhang S. · Department of Infectious Disease, The First Hospital of Xi'an Jiaotong University, Xi'an, China. · Am J Obstet Gynecol. · Pubmed #16458634 No free full text.

Abstract: OBJECTIVE: The purpose of this study was to investigate the presence and distribution of Hepatitis B virus antigen (HBVAg) and Hepatitis B virus deoxyribonucleic acid (HBVDNA) in ovary with chronic hepatitis B virus. STUDY DESIGN: The immunohistochemistry method and in situ hybridization method were used to detect the hepatitis B surface antigen (HBsAg), anti-hepatitis B core antibody (HBcAg), and HBVDNA in ovary of patients with chronic hepatitis B infection. RESULTS: HBsAg presented in the ovum-granular cell and interstitial cell of ovary. HBsAg located in the cytomembrane and cytoplasm. HBcAg presented in the ovum-granular cell-interstitial cell and endothelium cell of interstitial blood vessel of ovary, and the HBcAg may present in the ovum at different stage. HBcAg was situated at the cytomembrane, cytoplasm, and nucleus. HBVDNA was detected in the ovum at different stage as HBcAg and in granular cell, interstitial cell of ovary. In the same ovary tissue, HBVDNA was positive in some ova, but it was weakly positive or negative in other ova. HBVDNA distributed mainly in nucleus, but also can be detected in the cytoplasm. CONCLUSION: HBV could infect the ovum at different stage and replicate in it. This may be an important mechanism of HBV vertical transmission.

Tong X, Chase R, Skelton A, Chen T, Wright-Minogue J, Malcolm BA. · Department of Virology, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilwoth, NJ 07033, USA. · Antiviral Res. · Pubmed #16448708 No free full text.

Abstract: HCV NS3 protease variants resistant to the protease inhibitor SCH 503034 were selected. Three mutations, T54A, V170A and A156S mutations conferred low to moderate levels of resistance (<20-fold). Longer exposure (>10 passages) or selection with higher levels of compound led to the selection of a more resistant variant, A156T (>100-fold). [Lin, C., Lin, K., Luong, Y.P., Rao, B.G., Wei, Y.Y., Brennan, D.L., Fulghum, J.R., Hsiao, H.M., Ma, S., Maxwell, J.P., Cottrell, K.M., Perni, R.B., Gates, C.A., Kwong, A.D., 2004. In vitro resistance studies of hepatitis C virus serine protease inhibitors, VX-950 and BILN 2061: structural analysis indicates different resistance mechanisms. J. Biol. Chem. 279(17), 17508-17514; Lu, L., Pilot-Matias, T.J., Stewart, K.D., Randolph, J.T., Pithawalla, R., He, W., Huang, P.P., Klein, L.L., Mo, H., Molla, A., 2004. Mutations conferring resistance to a potent hepatitis C virus serine protease inhibitor in vitro. Antimicrob. Agents Chemother. 48(6), 2260-2266.] Combination with IFN-alpha drastically reduced the number of emergent colonies. Resistant colonies showed no change in sensitivity to IFN-alpha. Although the A156T mutation conferred the highest level of resistance to SCH 503034, it significantly reduced the colony formation efficiency (CFE) of the mutant replicon RNA, and rendered replicon cells less fit than those bearing wild-type replicons. Replicon cells bearing mutation A156S were less fit than wild-type in co-culture growth competition assays but showed no impact on CFE. The V170A mutation, on the other hand, did not affect replicon fitness in either assay, which was consistent with its emergence as the dominant mutant after 12 months of continuous selection. The reduced fitness of the most resistant variant suggests that it may be rare in naïve patients and that development of high-level resistance may be slow. Combination therapy with IFN-alpha should also greatly reduce the potential emergence of resistance.

Yi M, Tong X, Skelton A, Chase R, Chen T, Prongay A, Bogen SL, Saksena AK, Njoroge FG, Veselenak RL, Pyles RB, Bourne N, Malcolm BA, Lemon SM. · Center for Hepatitis Research, Institute for Human Infections & Immunity, University of Texas Medical Branch, Galveston 77555-1019, USA. · J Biol Chem. · Pubmed #16352601 links to  free full text

Abstract: Drug resistance is a major issue in the development and use of specific antiviral therapies. Here we report the isolation and characterization of hepatitis C virus RNA replicons resistant to a novel ketoamide inhibitor of the NS3/4A protease, SCH6 (originally SCH446211). Resistant replicon RNAs were generated by G418 selection in the presence of SCH6 in a dose-dependent fashion, with the emergence of resistance reduced at higher SCH6 concentrations. Sequencing demonstrated remarkable consistency in the mutations conferring SCH6 resistance in genotype 1b replicons derived from two different strains of hepatitis C virus, A156T/A156V and R109K. R109K, a novel mutation not reported previously to cause resistance to NS3/4A inhibitors, conferred moderate resistance only to SCH6. Structural analysis indicated that this reflects unique interactions of SCH6 with P'-side residues in the protease active site. In contrast, A156T conferred high level resistance to SCH6 and a related ketoamide, SCH503034, as well as BILN 2061 and VX-950. Unlike R109K, which had minimal impact on NS3/4A enzymatic function, A156T significantly reduced NS3/4A catalytic efficiency, polyprotein processing, and replicon fitness. However, three separate second-site mutations, P89L, Q86R, and G162R, were capable of partially reversing A156T-associated defects in polyprotein processing and/or replicon fitness, without significantly reducing resistance to the protease inhibitor.

Song Z, McClain CJ, Chen T. · Department of Medicine, University of Louisville Medical Center, Louisville, KY 40292, USA. · Pharmacology. · Pubmed #15240996 No free full text.

Abstract: An overdose of acetaminophen (APAP) is the most frequent cause of fulminant liver failure in the United States. Increasing evidence demonstrates that oxidative stress plays an important etiologic role in APAP-induced liver injury. S-Adenosylmethionine (SAMe) is a key intermediate in the hepatic trans-sulfuration pathway and serves as a precursor for glutathione (GSH) as well as the methyl donor in most transmethylation reactions. In the present study, we investigated effects of SAMe on liver injury induced by APAP administration in male C57BL/6 mice. Two related studies were performed. In the first experiment, SAMe (1g/kg BW) was injected intraperitoneally 4 h before APAP (600 mg/kg BW) administration. In the second experiment, SAMe was injected intraperitoneally 1 h after APAP administration. Our results showed that APAP administration induced changes typical of confluent centrilobular necrosis by histological examination and a marked elevation in serum alanine aminotransferase (ALT) activity. APAP administration induced significant decreases in both hepatic and blood SAMe concentrations. In addition, APAP decreased intracellular (both cytosolic and mitochondrial) GSH concentrations along with increased lipid peroxidation in conjunction with mitochondrial dysfunction as documented by Ca2+-induced mitochondrial permeability transition. SAMe treatment (both before and after APAP) significantly attenuated the liver injury. Exogenous SAMe prevented the decrease in liver and blood SAMe concentrations. Moreover, SAMe treatment attenuated both cytosolic and mitochondrial GSH depletion as well as mitochondrial dysfunction. We conclude that SAMe at least in part protects the liver from APAP-induced injury by preventing intracellular GSH depletion and mitochondrial dysfunction.

Deng T, Fan G, Chen T, Chen N, Hu D, Wang M, Jia K. · Institute of Liver Diseases, PLA, Beijing 100700, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #11953132 No free full text.

Abstract: OBJECTIVE: To develop a HCV combined HBV DNA-based therapeutic vaccine. METHODS: The HBV core gene and HCV core cDNA were inserted into the eukaryotic expression vector with two multiple cloning sites mammalian expression vector, which can be used for expression of two foreign genes, under the CMV promoter and RSC promoter respectively (named pRSC-HBV/HCV). Cellular expression of pRSC-HBV/HCV was assessed following transfection into SP2/0 cells. The Balb/c mice were immunized by multiple sites intramuscular injection with pRSC-HBV/HCV and the immune responses were detected. RESULTS: The 21-kd and 14-kd core protein was observed. Both anti-HBc IgG and anti-HCV core Ab were detected in all immunized mice. Strong CTL activity of splenocytes against SP2/0 cells expressing both HBV and HCV core proteins were measured in immunized mice both in vitro and in vivo. CONCLUSION: The investigation demonstrated that pRSC-HBV/HCV could generated both humoral immune response and Strong CTL activity against HBcAg and HCV nucleocapsid in mice.

Chen T, Luk JM, Cheung ST, Yu WC, Fan ST. · Center of Liver Diseases and Department of Surgery, University of Hong Kong Medical Center, Queen Mary Hospital, Pokfulam, Hong Kong. · J Clin Microbiol. · Pubmed #10790133 links to  free full text

Abstract: This study evaluated the applicability of quantitative PCR (Q-PCR) and branched-chain DNA assays for detection of hepatitis B virus (HBV) DNA in sera. For 42 samples, the detection rates were 81 and 41%, respectively, with a correlation coefficient of 0.633. The Q-PCR is useful for early monitoring of HBV load in high-risk patients.