慢性乙型肝炎乙肝病毒血清标志物检测研究

目的对慢性乙型肝炎患者进行乙肝病毒(HBV)血清标志物检测,分析结果,评价感染复制状况,选择适合临床防治需要、容易普及开展的标志物。方法采用酶联免疫吸附试验(ELISA)检测乙肝5项标志物[乙型肝炎病毒表面抗原(HBsAg)、乙型肝炎病毒表面抗体(抗-HBs)、乙型肝炎病毒e抗原(HBeAg)、乙型肝炎病毒e抗体(抗-HBe)、乙型肝炎病毒核心抗体(抗-HBc)]、前S1抗原、前S2抗原及前S2抗体,实时荧光定量检测HBV-DNA。286例慢性乙型肝炎患者分为HBeAg阳性组(126例)和HBeAg阴性组(160例),比较各组前S1抗原、前S2抗原、前S2抗体和HBV-DNA检出率。结果HBeAg阳性组,前S1抗原、前S2抗原、前S2抗体和HBV-DNA阳性率分别为:93.7%、77.0%、0和95.2%。HBeAg阴性组,前S1抗原、前S2抗原、前S2抗体和HBV-DNA阳性率分别为:55.0%、36.3%、0和58.1%。组内比较,HBeAg阳性组或HBeAg阴性组,前S1抗原与前S2抗原检出率差异具有统计学意义(P<0.05),前S1抗原与HBV-DNA检出率差异无统计学意义(P>0.05),前S2抗原与HBV-DNA检出率差异具有统计学意义(P<0.05)。组间相同项目比较,前S1抗原检出率、前S2抗原检出率、HBV-DNA检出率差异均具有统计学意义(P<0.05)。结论慢性乙型肝炎患者,前S1抗原与HBV-DNA具有良好的相关性,实验室常规开展乙肝5项和前S1抗原检测,可以判断乙肝病毒感染复制状况,为慢性乙型肝炎的预防、诊断和治疗提供实验依据。

肝病治疗仪合乙肝2号膏穴位敷贴治疗慢性乙型肝炎的观察

目的肝病治疗仪合乙肝2号膏穴位辨证敷贴透皮治疗慢性乙型肝炎疗效。方法将180例乙肝患者随机分为治疗组120例和对照组60例,治疗组为肝病治疗仪组(60例)和中药组(60例),对照组为干扰素组。治疗仪组和对照组均在常规保肝降酶治疗的基础上用药。结果治疗后3组在症状方面均得到明显改善,治疗组和中药组都优于干扰素组;而中药组在胁痛、乏力、尿黄等方面症状改善更明显,优于治疗仪组。在肝功能方面,3组ALT、AST、GGT均下降,且中药组明显优于干扰素组,而治疗仪组只在ALT方面明显优于干扰素组,且疗效没有中药组好。在肝纤维化方面,中药组与干扰素组之间无明显区别,而在HA、CIV治疗方面治疗仪组疗效不及中药组和干扰素组。在e抗原(HBeAg)转阴、e抗体(HBeAb)转阳、HBV-DNA滴度下降方面,干扰素组均优于治疗仪组和中药组。结论肝病治疗仪合乙肝2号穴位敷贴治疗慢性乙型肝炎,对改善临床症状及肝功能、减轻肝纤维化有较好的效果,而对抑制病毒复制无显著疗效。

慢性乙型肝炎病毒感染者健康知识调查

目的通过问卷调查了解公众对乙型肝炎防治知识的了解程度,探讨对乙肝人群健康教育的手段。方法采用问卷调查,问卷在患者就诊时自己填写,填写有困难者医生诊察过程中帮助填写,填写时以近2年的情况为依据,内容均真实可靠。计数资料采用χ2检验确定差异性。结果大部分HBV感染者希望对肝炎知识有所了解,83.83%的HBV感染者希望了解全部或大部分肝炎知识,了解肝炎知识的途径主要有电视21.84%（199/911）、报纸杂志18.77%（171/911）、肝炎书籍18.00%（164/911）、亲朋病友介绍15.26%（139/911）、医生介绍11.96%（109/911）、电台广播9.44%（86/911）、肝炎知识讲座2.41%（22/911）、其他途径7.79%（71/911）。肝炎知识更新较慢,28.10%的人12个月以上才了解一次肝炎知识,HBV感染者中48.96%认为系垂直传播,仍有9.77%的人认为乙肝疫苗不能预防肝炎传染。HBV感染后对患者的生活习惯和劳动能力构成了一定影响,病情愈重影响愈大,27.18%的人1-3个月内进行一次血清学检查,58.2%的HBV感染者接受过治疗。结论公众对肝炎知识的了解较少,知识更新较慢,部分人对HBV感染重视不够,而另一部分人存在过度治疗。

拉米夫定治疗慢性乙型肝炎52例疗效观察

目的观察拉米夫定治疗慢性乙型肝炎的临床疗效。方法52例患者口服拉米夫定,100mg/d,疗程12个月,治疗过程中不能使用其他抗HBV药、降酶药、"保肝药"、免疫调节剂等。治疗12个月后测患者血清HBV-DNA水平。结果治疗前血清HBV-DNA平均水平6.82±1.31,治疗12个月后3.21±0.98。治疗12个月后HBV-DNA阳性例数21例,阳性率37.5%,治疗前100%。结论拉米夫定治疗慢性乙型肝炎临床疗效确切,值得临床应用。

拉米夫定联合胸腺肽、苦参素治疗慢性HBeAg阴性乙型肝炎疗效观察

目的对比观察拉米夫定联合胸腺肽、苦参素与单用拉米夫定治疗HBeAg阴性慢性乙型肝炎的持久治疗效果。方法设拉米夫定联合胸腺肽、苦参素为治疗组，单用拉米夫定为对照组。对比观察两组治疗效果。结果治疗结束时联合治疗组完全应答率93．3％。单用拉米夫定组60％，两组比较P〈0．05，随访6个月治疗组完全应答率76．6％．对照组33．3％，P〈0.01。随访一年时完全应答率为53．3％，对照组完全应答率23．3％，P〈0．025。两组完全应答率比较差异显著。随访1年时治疗组肝纤维化指标正常率53．3％，对照组肝纤维化指标正常率23.3％，P〈0．025。结论拉米夫定联合胸腺肽、苦参素治疗能提高对HBeAg阴性慢性乙型肝炎的持久治疗效果。

柴贝四逆饮治慢性乙型肝炎100例分析

目的:观察柴贝四逆饮对慢性乙型肝炎患者肝功能的影响。方法:100例患者随机分为治疗组和对照组各50例,对照组用甘草酸二铵注射液,硫普罗宁注射液,复方丹参注射液静脉点滴,治疗组在对照组基础上服柴贝四逆饮,并测定两组治疗前后肝功能水平。结果:治疗组肝功能正常率86%,显著优于对照组62%(P<0.05)治疗组复发率13.33%显著少于对照组35.71%(P<0.05)。结论:柴贝四逆饮能显著改善慢性乙型肝炎患者的肝功能。

强肝解毒汤配合苦参素治疗慢性乙型肝炎60例

目的:比较强肝解毒汤配合苦参素与苦参素治疗慢性乙型肝炎的临床疗效。方法:将患者随机分为治疗组(强肝解毒汤配合苦参素)60例和对照组(苦参素)40例,对照组:苦参素注射液0.6g,加液静滴,1次/d,疗程2个月,继而口服苦参素胶囊0.2g,3次/d,疗程4个月,总疗程6个月;治疗组:在对照组的基础上,给予强肝解毒汤,每日1剂,总疗程6个月。观察治疗前后患者的肝功能、血清病毒标志物等指标。结果:治疗组与对照组比较,治疗前后患者的肝功能(ALT、AST、TBil)的改善及HB VDNA阴转,均有显著性差异(P<0.05)。结论:强肝解毒汤配合苦参素治疗慢性乙型肝炎,特别对患者肝功能的恢复及HB VDNA的转阴具有较好的临床疗效,且费用低廉,无毒副反应,其作用机制有待进一步研究。

自拟复肝汤对慢性乙肝病毒蛋白表达的影响

应用自拟复肝汤治疗慢性乙肝136例,从血清观察自拟复肝汤抗病毒的疗效。治疗前、治疗后分别检测血清HBVDNA(PCR定性、定量)和血清HBVM(ELISA法)。治疗后病毒含量减少,血清HBeAg治疗后6个月阴转率30%。自拟复肝汤对慢性乙型肝炎有一定的抗病毒作用。

Role of Antivirus Therapy in Treatment of Hepatocellular Carcinoma with Chronic Hepatitis B Infection

Objective： To observe the recurrence and prognosis of hepatocellular carcinoma （HCC） patients coexisting with chronic hepatitis B infection with active virus replication after receiving antivirus therapy using lamivudine and thymosin α1 （Tα1） postoperatively. Methods： From Jan. 2000 to Dec. 2003, 70 patients with HCC coexisting chronic hepatitis B infection with active virus replication were prospectively divided into two groups： control group （n=35） received hepatectomy only; treatment group （n=35） received hepatectomy and lamivudine plus Tα1 therapy postoperatively. The suppression of HBV-DNA, HBeAg seroconverted rate, tumor recurrent rate and the median survival for the two groups were observed and calculated. Results： In treatment group and control group, the 2-year HBV-DNA suppression rate was 100% vs. 4% （P=0.0000）; HBeAg seroconverted rate was 73.0% vs. 7.5% （P〈0.05）; the recurrent rate was 10.0 vs 6.5 months （P=0.0032）; the median survival time was 12.5 vs. 6.0 months （P=0.0023）, respectively. Conclusion： Antivirus therapy using lamivudine and Tα1 postoperatively may suppress the HBV reaction, delay the recurrent time and prolong the survival for HCC patients coexisting chronic HBV infection with active virus replication.

Reduced bone mineral density and altered bone turnover markers in patients with non-cirrhotic chronic hepatitis B or C infection

AIM: Previous studies suggest that loss of bone mineral density (BMD) frequently occurs in patients with chronic viral liver disease, presenting with histologically proven liver cirrhosis. However, little is known about the occurrence of bone disease in non-cirrhotic patients with chronic hepatitis B or C. Therefore, it was the aim of this study to evaluate this particular population for BMD and bone turnover markers. METHODS: Biochemical markers of bone turnover and BMD were measured in 43 consecutive patients with HCV (n = 30) or HBV (n = 13) infection without histological evidence for liver cirrhosis. Mean age was 49 years (range 26-77 years). BMD was measured by dual X-ray absorptiometry in the femoral neck (FN) and the lumbar spine (LS) region. In addition, bone metabolism markers were measured. RESULTS: BMD was lowered in 25 (58%) of the patients with chronic hepatitis B or C (FN; 0.76 (0.53-0.99); LS: 0.96 (0.62-1.23) g/cm2). Eight (32%) osteopenic patients were diagnosed with osteoporosis. Bone-specific alkaline phosphatase (P= 0.005) and intact parathyroid hormone (iPTH) (P = 0.001) were significantly elevated in the more advanced stages of fibrosis. Mean T-score value was lower in patients with chronic hepatitis C as compared to patients suffering from chronic hepatitis B; however, the difference was not statistically significant (P= 0.09). CONCLUSION: There was a significantly reduced BMD in non-cirrhotic patients with chronic hepatitis B or C infection. Alterations of bone metabolism already occurred in advanced liver fibrosis without cirrhosis. According to our results, these secondary effects of chronic viral hepatitis should be further investigated.

Development of Novel Therapeutics for Chronic Hepatitis B

Chronic infection of hepatitis B virus (HBV) presents one of the serious public health challenges worldwide. Current treatment of chronic hepatitis B (CHB) is limited, and is composed of interferon and nucleoside/nucleotide reverse transcriptase inhibitors (NRTI). Interferon is poorly tolerated and is only responsive in a small fraction of CHB patients and NRTIs often face the problem of emergence of drug resistance during long-term treatment. The current treatment of CHB can be improved in several ways including genotyping mutations associated with drug resistance before treatment to guide the choice of NRTIs and suitable combinations among NRTIs and interferon. It is important to continue research in the identification of novel therapeutic targets in the life cycle of HBV or in the host immune system to stimulate the development of new antiviral agents and immunotherapies. Several antiviral agents targeting HBV entry, cccDNA, capsid formation, viral morphogenesis and virion secretion, as well as two therapeutic vaccines are currently being evaluated in preclinical studies or in clinical trials to assess their anti-HBV efficacy.

Polymorphisms of some cytokines and chronic hepatitis B and C virus infection

AIM： To study the relationship between the polymorphisms in some cytokines and the outcome of hepatitis B virus （HBV） and hepatitis C virus （HCV） infection. METHODS： Samples were obtained from 203 patients infected with HBV and/or HCV while donating plasma in 1987, and 74 controls were obtained from a rural area of North China. Antibodies to HBV or HCV antigens were detected by enzyme-linked imrnunoassay. The presence of viral particles in the serum was determined by nested reverse-transcriptase polymerase chain reaction （PCR）. Hepatocellular injury, as revealed by alanine aminotransferase （ALT） and aspartate aminotransferase level, was detected by a Beckman LX-20 analyzer. DNA was extracted from blood cells. Then, the single nucleotide polymorphisms of IL-2-330, IFN-γ＋874, IL-10-1082/-592 and IL-4-589 were investigated by restriction fragment length polymorphism-PCR or sequence specific primer-PCR.RESULTS： Persistent infection with HBV, HCV, and HBV/HCV coinfection was associated with IL-2-330 TT genotype and T allele, IFN-γ＋874 AA genotype, and IL-10-1082 AA genotype. The clinical outcome of HBV and/or HCV infection was associated with IL-2-330 TT genotype and T allele, IFN-γ＋874 AA genotype, and IL-10-1082 AA genotype. IL-2-330 GG genotype frequency showed a negative correlation with clinical progression, IL-10-1082 AA genotype frequency showed a positive correlation and IL-10-1082 AG genotype frequency showed a negative correlation with clinical progression. HCV RNA positive expression was associated with IL-10-1082 AA genotype and the A allele frequency. Abnormal serum ALT level was associated with IL-10-592 AC genotype frequency and IL-4-589 CC genotype, CT genotype, and the C allele. CONCLUSION： These results suggest that polymorphisms in some cytokine genes influence persistent HBV and HCV infection, clinical outcome, HCV replication, and liver damage.

Psychometrics of the chronic liver disease questionnaire for Southern Chinese patients with chronic hepatitis B virus infection

AIM： To test the psychometric properties of a Chinese [（Hong Kong） HK] translation of the chronic liver disease questionnaire （CLDQ）. METHODS： A Chinese （HK） translation of the CLDQ was developed by iterative translation and cognitive debriefing. It was then administered to 72 uncomplicated and 78 complicated chronic hepatitis B （CriB） patients in Hong Kong together with a structured questionnaire on service utilization, and the Chinese （HK） SF-36 Health Survey Version 2 （SF-36v2）. RESULTS： Scaling success was ≥ 80% for all but three items. A new factor assessing sleep was found and items of two （Fatigue and Systemic Symptoms） subscales tended to load on the same factor. Internal consistency and test-retest reliabilities ranged from 0.58-0.90 for different subscales. Construct validity was confirmed by the expected correlations between the SF-36v2 Health Survey and CLDQ scores. Mean scores of CLDQ were significantly lower in complicated compared with uncomplicated CHB, supporting sensitivity in detecting differences between groups.CONCLUSION： The Chinese （HK） CLDQ is valid, reliable and sensitive for patients with CHB. Some modifications to the scaling structure might further improve its psychometric properties,

Replication of clinical hepatitis B virus isolate and its application for selecting antiviral agents for chronic hepatitis B patients

AIM: To establish a cell model harboring replicative clinical hepatitis B virus (HBV) isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B (CHB) patients. METHODS: The full-length HBV genomic DNA from 8 CHB patients was amplified by polymerase chain reaction (PCR). All the patients were treated with lamivudine for at least seven months and finally became resistant to lamivudine. The amplified HBV DNA fragments were inserted into pHY106 vectors by Sap Ⅰ?digestion. The recombinant plasmids containing 1.1 copies of HBV genome were transiently transfected into Huh7 cell line, and the levels of HBsAg, HBeAg and intercellular HBV replicative intermediates were determined by ELISA and Southern blot analysis, respectively, with or without lamivudine and adefovir treatment. The antiviral treatment with adefovir was administered to the patients and analyzed in parallel. RESULTS: A total of 25 independent HBV isolateswere obtained from the sera of 8 patients, each patient had at least two isolates. One isolate from each individual was selected and subcloned into pHY106 vector, including 5 isolates with YVDD mutation and 3 isolates with YIDD mutation. All recombinant plasmids harboring HBV isolates were transfected into Huh7 cells. The results indicated that HBV genome carried in HBV replicons of clinical HBV isolates could effectively replicate and express in Huh7 cells. Adefovir, but not lamivudine, inhibited HBV replication both in vitro and in vivo, and in vitro inhibition was dose-dependent. CONCLUSION: The novel method described herein enables individualized selection of anti-HBV agents in clinic and is useful in future studies of antiviral therapy for CHB.

Assessment of health-related quality of life in Chinese patients with minimal hepatic encephalopathy

AIM:To evaluate the health-related quality of life (HRQOL) based on the Chinese version of SF-36 and Chronic Liver Disease Questionnaire (CLDQ) in subjects with chronic hepatitis B,liver cirrhosis,including patients with minimal hepatic encephalopathy (MHE). METHODS:The SF-36 and CLDQ were administered to 160 healthy volunteers,20 subjects with chronic hepatitis B and 106 patients with cirrhosis (33 cases exhibited MHE). HRQOL scores were compared among the different study groups. The SF-36 includes eight health concepts:physical functioning,role-physical,body pain,general health,vitality,social functioning,role-emotion,and mental health. Six domains of CLDQ were assessed:abdominal symptoms,fatigue,systemic symptoms,activity,emotional function and worry. RESULTS:Compared with healthy controls (96.9 ± 4.5,86.6 ± 18.4,90.1 ± 12.5,89.0 ± 5.7,87.5 ± 4.3,95.8 ± 7.1,88.5 ± 15.9,88.7 ± 5.2 in SF-36 and 6.7 ± 0.5,6.1 ± 0.6,6.3 ± 0.6,6.5 ± 0.5,6.3 ± 0.5,6.8 ± 0.4 in CLDQ),patients with chronic hepatitis B (86.3 ± 11.0,68.8 ± 21.3,78.9 ± 14.4,60.8 ± 10.5,70.8 ± 8.6,76.1 ± 12.6,50.0 ± 22.9,72.2 ± 10.6 and 5.5 ± 1.0,4.5 ± 1.0,5.2 ± 1.1,5.3 ± 0.9,4.8 ± 0.9,4.9 ± 1.0) and cirrhosis (52.8 ± 17.4,32.8 ± 27.9,61.6 ± 18.9,30.2 ± 18.3,47.9 ± 20.1,54.0 ± 19.2,28.9 ± 26.1,51.1 ± 17.8 and 4.7 ± 1.2,3.9 ± 1.2,4.7 ± 1.2,4.7 ± 1.3,4.7 ± 1.0,4.4 ± 1.1) had lower HRQOL on all scales of the SF-36 and CLDQ (P < 0.01 for all). Increasing severity of liver cirrhosis (based on the Child-Pugh score/presence or absence of MHE) was associated with a decrease in most components of SF-36 and CLDQ,especially SF-36.CONCLUSION:The Chinese version of SF-36 along with CLDQ is a valid and reliable method for testing MHE in patients with liver cirrhosis. Cirrhosis and MHE are associated with decreased HRQOL.

Development of hepatitis C virus vaccine using hepatitis B core antigen as immuno-carrier

AIM: To develop hepatitis C virus (HCV) vaccine using HBcAg as the immuno-carrier to express HCV T epitope and to investigate its immunogenicity in mice. METHODS: We constructed the plasmid pTrc-coreNheI using gene engineering technique, constructed the pcDNA3.1-coreNheI-GFP plasmid with GFP as the reporter gene, and transfected them into Hela cells. The expression of GFP was observed under confocal microscopy and the feasibility of using HBcAg as an immuno-carrier vaccine was studied. pTrc-core gene with a synthetic T epitope antigen gene of HCV (35-44aa) was fused and expressed in the plasmid pTrc- core-HCV (T). For the fusion of the HBcAg-T protein, sucrose, density gradient centrifugation was used, and its molecular weight and purity were analyzed by SDS- PAGE. Then balb/c mice were immunized by the plasmid with the HBcAg (expressed by pTrc-core) protein as control. The tumor regression potential was investigated in mice and evaluated at appropriate time. After three times of immunization, the peripheral blood and spleen of vaccinated mice were collected. HBcAb was detected by ELISA, and nonspecific T lymphocyte proliferation and response of splenocytes were respectively examined by MTT assay. T cell subset of blood and spleen were detected by FACS. RESULTS: GFP was successfully expressed. Tumor regression trial showed that no tumor formation was found in the group receiving immunization, while tumor xenograft progression was not changed in the control group. Strong nonspecific lymphocyte proliferation response was induced. FACS also showed that the ratio of CD8+ T cells in the experimental group was higher than the controls, but the serum HBcAb in experimental group was similar to the control. CONCLUSION: HBcAg can be used as an immuno-carrier of vaccine, the fusion of HBcAg-T protein could induce stronger cellular immune responses and it might be a candidate for therapeutic vaccines specific for HCV.

Association between TRAIL expression on peripheral blood lymphocytes and liver damage in chronic hepatitis B

AIM:To explore a novel mechanism for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), upregulation of CD4+ and CD8+T lymphocytes participating in the patho-physiological process of chronic hepatitis B (CHB). METHODS: The levels of serum soluble TRAIL (sTRAIL), serum IFN-γ and membrane-bound TRAIL expression on peripheral leucocytes from 58 CHB patients were examined by ELISA and flow cytometry respectively. The levels of TRAIL were compared with the baseline levels of 17 healthy controls, and correlation analysis was performed between ALT, TBIL, PT, morphological change in hepatic tissues, and serum IFN-γ. RESULTS: The results showed that TRAIL levels on membranes of CD4+, CD8+ T cells in CHB patients were much higher than those in healthy controls (P<0.001), and were correlated with serum TBIL (r=0.354, P= 0.008 for CD4+ and r= 0.522, P= 0.000 for CD8+, respectively), ALT (r= 0.393, P= 0.003 for CD8+), PT (r = 0.385, P = 0.004 for CD8+) and serum IFN-y level (r = 0.302, P= 0.011 for CD4+ and r= 0.307, P= 0.009 for CD8+). On the contrary to membrane-bound TRAIL expression, serum level of sTRAIL was not correlated with that of TBIL and PT, though it was higher than that of the normal population and was positively correlated with serum HBeAg expression (r= 0.695, P = 0.001). CONCLUSION: The expression level of TRAIL on the membrane of lymphocytes was upregulated and associated with the liver injury in CHB patients. These findings suggest that upregulation of TRAIL expression may be induced by virus antigen and inflammatory cytokine IFN-γ.

A combination treatment of entecavir and early-phase corticosteroid in severe exacerbation of chronic hepatitis B

Of patients with severe exacerbation of chronic hepatitis B accompanied by jaundice and coagulopathy,20%-30%have a fatal outcome.In this report,we describe 2 cases of severe exacerbation of chronic hepatitis B with jaundice and coagulopathy who were successfully treated with a combination of entecavir and corticosteroid.In both cases,rapid reductions in serum hepatitis B virus(HBV)-DNA levels were observed,and corticosteroid was stopped after serum HBV-DNA levels became undetectable.Entecavir treatment was continued.Generally,entecavir treatment reduced serum HBV-DNA levels rapidly,although the improvement in liver function was delayed by a few weeks.During this time lag,liver cell injury continued and the disease progressed.Corticosteroid suppressed the excessive host immune response and was useful for stopping progressive deterioration.A combination of entecavir and early-phase corticosteroid may be a useful treatment in severe exacerbation of chronic hepatitis B.

Long-term alpha interferon and lamivudine combination therapy in non-responder patients with anti-HBe-positive chronic hepatitis B:Results of an open,controlled trial

AIM: To investigate the safety and efficacy of long-term combination therapy with alpha interferon and lamivudine in non-responsive patients with anti-HBe-positive chronic hepatitis B.METHODS: 34 patients received combination treatment (1 month lamivudine, 12 month lamivudine+interferon, 6month lamivudine), 24 received lamivudine (12 months),24 received interferon (12 months). Interferon was administered at 6 MU tiw and lamivudine at 100 mg orally once daily. Patients were followed up for 6 months after treatment.RESULTS: At the end of treatment, HBV DNA negativity rates were 88 % with lamivudine+interferon, 99 % with lamivudine and 55 % with interferon, (P=0.004, combination therapy vs. interferon, and P=0.001 lamivudine vs.interferon), and serum transaminase normalization rates were 84 %, 91% and 53 % (P=0.01 combination therapy vs. interferon, and P=0.012 lamivudine vs. interferon). Six months later, HBV DNA negativity rates were 44 % with lamivudine+interferon, 33 % with lamivudine and 25 % with interferon, and serum transaminase normalization rates were 61%, 42 % and 45 %, respectively, without statistical significance. No YMDD variants were observed with lamivudine+interferon (vs. 12 % with lamivudine). The combination therapy appeared to be safe. CONCLUSION: Although viral clearance and transaminase normalization are slower with long-term lamivudine+interferon than that with lamivudine alone, the combination regimen seems to provide more lasting benefits and to protect against the appearance of YMDD variants. Studies with other regimens regarding sequence and duration are needed.

Mechanism of T cell hyporesponsiveness to HBcAg is associated with regulatory T cells in chronic hepatitis B

AIM： To study the mechanisms of hyporesponsiveness of HBV-specific CD4^＋ T cells by testing TH1 and TH2 commitment and regulatory T cells. METHODS： Nine patients with chronic hepatitis B were enrolled. Peripheral blood mononuclear cells were stimulated with HBcAg or HBsAg to evaluate their potential to commit to TH1 and TH2 differentiation. HBcAg-specific activity of regulatory T cells was evaluated by staining with antibodies to CD4, CD25, CTLA-4 and interleukin-10. The role of regulatory T cells was further assessed by treatment with anti-interleukin-10 antibody and depletion of CD4^＋CD25^＋ cells. RESULTS： Level of mRNAs for T-bet, IL-12R β2 and IL-4 was significantly lower in the patients than in healthy subjects with HBcAg stimulation. Although populations of CD4^＋CD25^highCTLA-4^＋ T cells were not different between the patients and healthy subjects, IL-10 secreting cells were found in CD4^＋ cells and CD4^＋CD25^＋ cells in the patients in response to HBcAg, and they were not found in cells which were stimulated with HBsAg. Addition of anti-IL-10 antibody recovered the amount of HBcAgspecific TH1 antibody compared with control antibody （P 〈 0.01, 0.34% ± 0.12% vs 0.15% ± 0.04%）. Deletion of CD4^＋CD25^＋ T cells increased the amount of HBcAgspecific TH1 antibody when compared with lymphoo/tes reconstituted using regulatory T cells （P 〈 0.01, 0.03% ± 0.02% vs 0.18% ± 0.05%）.CONCLUSION： The results indicate that the mechanism of T cell hyporesponsiveness to HBcAg includes activation of HBcAg-induced regulatory T cells in contrast to an increase in TH2-committed cells in response to HBsAg.