晶珠肝泰舒治疗慢性乙型肝炎60例疗效观察

目的观察晶珠肝泰舒治疗慢性乙型肝炎的疗效.方法对60例确诊为慢性乙型肝炎患者,给予晶珠肝泰舒胶囊,每次2粒,每日3次,3月为一疗程,观察症状、体征、肝功能及HBV标志物治疗前后的变化.结果晶珠肝泰舒胶囊能明显改善症状及肝功能.治疗前后平均ALT分别为(175.5±93.7)U/L和(49.5±43.6)U/L,AST分别为(102.9±98.4)U/L和(47.1±39.8)U/L,血清总胆红素分别为(51.3±47.2)μmol/L和(19.64±9.5)μmol/L,P值均＜0.01. HBsAg阴转3例,HBeAg 8/27例(29.6%)阴转,HBV DNA 11/31例(35.4%)阴转.结论晶珠肝泰舒胶囊具有保肝、降酶、退黄的良好作用和一定的抗乙型肝炎病毒作用.     

晶珠肝泰舒胶囊长期毒性试验

连续大剂量(成人用量的300倍,100倍)、较长疗程(45d,90d)灌服晶珠肝泰舒胶囊,大鼠体重明显增长,对大鼠心、肝、脾、肺、肾、肾上腺、胸腺均无明显毒性损害,表明大剂量、长疗程服用晶珠肝泰舒胶囊毒性低,有促进生长作用.     

晶珠肝泰舒对大鼠实验性肝损伤的影响

目的研究藏药晶珠肝泰舒对实验性纤维化大鼠的影响。方法 60只Wistar大鼠随机分为正常对照组、模型组、联苯双酯滴丸对照组和晶珠肝泰舒高、中、低剂量组,采用CCl4诱导大鼠肝纤维化模型,晶珠肝泰舒高、中、低剂量组和联苯双酯滴丸组的大鼠自造模次日起灌胃给药,连续42 d,测定肝脏中羟脯氨酸、肝胶原蛋白和血清中血脂的含量。结果晶珠肝泰舒能降低肝纤维化大鼠肝脏中羟脯氨酸、肝胶原蛋白及血清中TCH、TG含量。结论晶珠肝泰舒对大鼠的实验性纤维化有显著疗效。     

薄层扫描法测定藏药晶珠肝泰舒胶囊中齐墩果酸的含量

目的　用薄层扫描法对晶珠肝泰舒胶囊中齐墩果酸进行定量分析。方法　反射法锯齿扫描 ,λ=5 2 0nm,线性化参数 Sx=3。展开剂 :甲苯 -醋酸乙酯 -冰醋酸 ( 1 2∶ 4∶ 0 .5 )。结果　齐墩果酸标准曲线线性范围为 1 .0～ 5 .0 μg( r=0 .9997) ,平均回收率为 97% ,RSD为 1 .5 9% ( n =4)。结论　方法准确 ,回收率好     

山芝麻水提液对HepG2.2.15细胞HBsAg和HBeAg的抑制作用

目的观察山芝麻水提液（water extracts from Helicteres angustifolia,WHA）的体外抗乙型肝炎病毒的作用。方法采用HepG2．2．15细胞模型进行体外培养,给予不同浓度山芝麻,以拉米夫定作阳性对照,作用72h后检测上清液中HBsAg、HBeAg的分泌,观察药物对HepG2．2．15细胞分泌HBV病毒抗原的影响,同时以MTT法检测药物在体外对HepG2．2．15细胞的生长抑制作用,从而评价药物的抗HBV作用。结果山芝麻水提液对HepG2．2．15细胞的半数细胞毒浓度（TC50）为482．1mg·L-1;对HepG2．2．15细胞分泌HBsAg的半数抑制浓度（IC50）为7．3mg。L-1,其治疗指数（TI）为66;对HBeAg,其Ic,n为14．6mg·L-1,TI为33。结论山芝麻水提液在体外有显著的抗HBV的作用,且毒性较低。Tel：（0771）5358342E—mail：gxLx60@163．COll]     

复方六月雪对HepG2.2.15细胞HBsAg和HBeAg的抑制作用

目的：观察复方六月雪（CLYX）体外抗乙型肝炎病毒（HBV）的作用。方法：采用四甲基噻唑蓝（MTT）法检测CLYX对HepG2．2．15细胞的半数毒性浓度（TC50）和最大无毒浓度（TG0）；在TG0基础上观察不同浓度药物作用于HepG2．2．15N胞，分别在第72h和144h收集细胞培养上清液，采用酶联免疫吸附实验（ELISA）法测定上清液HBsAg和HBeAg的滴度。结果：TG50为3．070g·L^-1，TG，为0．945g·L^-1，复方六月雪对HepG2．2．15N胞毒性较低。无毒浓度的复方六月雪在HepG2．2．15N胞培养中可有效地抑制细胞HBsAg（乙型肝炎表面抗原）和HBeAg（乙型肝炎E抗原）的分泌；且治疗指数（TI）均大于2，为高效低毒的抗HBV药物。结论：CLYX在体外有显著的抗HBV的作用，且毒性较低。     

空心莲子草总皂苷对HepG2.2.15细胞系HBsAg和HBeAg表达及HBV-DNA含量的影响

目的：研究空心莲子草总皂苷对乙肝病毒的抑制作用。方法：应用光密度测定法和细胞培养技术,研究空心莲子草总皂苷对HepG2.2.15细胞系HBsAg与HBeAg表达的影响。结果：96孔板试验：实验第3、6日,空心莲子草总皂苷对HBsAg和HBeAg表达均有明显的抑制作用（与对照组比较P<0.01）;同时采用定量PCR方法检测,空心莲子草总皂苷（200 mg·L-1）可使培养基中HBV-DNA转阴。结论：空心莲子草总皂苷有较强的体外抗乙肝病毒作用。     

荔枝核提取物对HepG 2.2.15细胞系HBsAg与HBeAg表达的影响

目的:研究荔枝核对乙型肝炎病毒的抑制作用及其有效部位.方法:应用HepG 2.2.15细胞系培养系统检测荔枝核提取物A、B、C、D、E、F对HBsAg与HbeAg表达的影响.结果:荔枝核提取物A、B、C、D、E、F(200,100mg·L-1)对HBsAg 和HbeAg表达均有抑制作用,其中E成分作用最强,在200 mg·L-1的浓度下,于实验第3天对HBsAg的抑制率为50%,对HBeAg的抑制率为20%,于实验第9天对HBsAg的抑制率为90.9%,对HBeAg的抑制率为84.3%(与对照组比较P＜0.01).结论:荔枝核提取物体外有较强的抗乙肝病毒作用.     

对乙型肝炎病毒HBsAg和HBeAg的抑制作用

目的 观察从广西藤茶中提取出的一有效部位TTF抗乙型肝炎病毒的作用. 方法 采用MTT法观察TTF含药血清对2.2.15细胞分泌HBsAg、HBeAg的抑制作用. 结果 TTF含药血清对2.2.15细胞分泌的HBsAg、HBeAg有显著抑制作用. 结论 TTF可明显抑制2.2.15细胞HBsAg、HBeAg的分泌,提示TTF具有抗HBV的作用.     

晶珠泰舒治疗慢性乙型肝炎45例疗效观察

目的：观察晶珠肝泰舒胶囊治疗慢性乙型肝炎的疗效，方法：选我院2000年6月至9月收住院的45例慢性乙型肝炎（中度或重度）患者。患者均有明显的消化道症状及乏力等。治疗前全部病例血清HBVDNA阳性，其中HBsAg阳性者35例。无其它肝炎病毒重叠感染。全部病例均接受晶球肝泰舒胶囊治疗。采用2粒／次，3次／日，治疗一疗程（三个月）并观察临床症状、体征、化验检查及融反应。结果：总有效率93.3%，HBVDNA阴转率10％，HBeAg阴转率36.3%，病例均未见明显的副反应。结论：晶珠肝泰舒具有明显的护肝作用，且有一定的抗病毒作用，其口服给药方便、廉价，值得进一步推广。     

核酸类似物PNA在HepG2．2．15细胞中对乙型肝炎病毒复制的抑制作用

目的评价核酸类似物PNA在HepG2．2．15细胞中对乙型肝炎病毒复制的抑制作用。方法以HepG2．2．15细胞作为细胞模型,拉米夫定作为阳性对照药物。HepG2．2．15细胞于药物处理14d后,收集上清液及细胞。采用ELISA检测上清液中HBsAg和HBeAg含量,HBV荧光定量检测上清液和细胞内HBV DNA水平,CCK-8试剂盒检测药物对细胞的毒性作用。结果核酸类似物PNA与拉米夫定在浓度1．6-1000μmol·L^-1内对HepG2．2．15细胞的毒性均较小。与药物未处理组比较,PNA和拉米夫定均可有效抑制细胞上清HBV DNA,半数抑制浓度（IC50）分别为0．05-0．10μmol·L^-1和0．09-0．18μmol·L^-1,两者之间差异无统计学意义。结论在体外细胞实验中,PNA对细胞的毒性小,与拉米夫定的安全指数相当;PNA对乙型肝炎病毒复制有较强的抑制作用,且具有一定时效量效关系,与拉米夫定的抑制强度相当。     

Maturation of recombinant hepatitis B virus surface antigen particles

The major surface antigen of Hepatitis B virus (HBsAg) is a cysteine-rich, lipid-bound protein with 226 amino acids. Recombinant HBsAg (rHBsAg) with associated lipids can self-assemble into 22-nm immunogenic spherical particles, which are used in licensed Hepatitis B vaccines. Little is known about the structural evolvement or maturation upon assembly beyond an elevated level of disulfide formation. In this paper, we further characterized the maturation of HBsAg particles with respect to their degree of cross-linking, morphological changes, and changes in conformational flexibility. The lipid-containing rHBsAg particles undergo KSCN- and heat-induced maturation by formation of additional intra- and inter-molecular disulfide bonds. Direct measurements with atomic force microscopy (AFM) revealed morphological changes upon maturation through KSCN-induced and heat-/storage-incurred oxidative refolding. Particle uniformity and regularity was greatly improved, and protrusions formed by the protein subunits were more prominent on the surface of the mature particles. Decreased conformational flexibility in the mature rHBsAg particles was demonstrated by millisecond-scale unfolding kinetics in the presence of an environment-sensitive conformation probe. Both the accessible hydrophobic cavities under native conditions and the changeable hydrophobic cavities upon denaturant-induced unfolding showed substantial decrease upon maturation of the rHBsAg particles. These changes in the structural properties may be critical for the antigenicity and immuno-genicity of this widely-used vaccine component.     

乙型肝炎表面抗原定量与原发性肝癌的相关性研究

目的 比较慢性乙型肝炎(CHB)与HBV相关性原发性肝癌患者中乙肝表面抗原(HBsAg)和HBV DNA 载量情况。方法 采用化学发光法检测403例CHB及肝细胞肝癌(HCC)患者血清乙型肝炎病毒标志物滴度,采用荧光定量PCR技术检测患者血清HBV DNA载量。403例患者按照临床诊断分为CHB组(209例)和HCC组(194例)。结果 CHB组:血清HBsAg 滴度≥250 IU/mL 者占89.00%; HBV DNA≥1 000 copies /mL 者占88.40%。HCC 组: 血清HBsAg 滴度≥250 IU/mL 者占79.90%; HBV DNA≥1 000 copies /mL 者占67.70%。两组HBsAg≥250 IU/mL患者的比例差异、HBV DNA≥1 000 copies/mL 患者的比例差异均有统计学意义(P<0.05)。结论 与CHB相比,HBV相关性原发性肝癌患者HBsAg、HBV DNA载量较低。     

Role of silent hepatitis B virus in chronic hepatitis B surface antigen(-) liver disease.

Despite a number of studies documenting hepatitis B virus (HBV) infection in the absence of hepatitis B surface antigen (HBsAg) a causal relationship between silent HBV infection and liver disease remain difficult to establish. In particular, both the prevalence and clinical significance of this observation are poorly understood. Why is HBV replication apparently so low in these patients? A number of studies have tried to elucidate the mechanism of HBsAg negative infections, and considerable data documenting HBV infectivity or reinfection in the absence of detectable HBsAg support the hypothesis that in some of these cases, HBV is undergoing low-level replication in the liver and this, in several situations including: (1) chronic liver disease, alcoholic liver disease, hepatocellular carcinoma; (2) viral reactivation following cancer chemotherapy or immunosuppression and (3) transmission via transfusion or from human serum to chimpanzees. In a recent study including 50 patients with chronic liver disease of unknown etiology we could detect serum HBV DNA by nested polymerase chain reaction (PCR) in 15/50 patients (50% at the cirrhosis stage) in the absence of HBsAg; in the liver of the 15 patients both HBcAg and/or HBsAg can be detected at very low-level. Viral host factors allowing HBV persistence in the absence of HBsAg can depend on several mechanisms. Coinfections with HCV can explain only a proportion of HBsAg(-) HBV infections. Secondly, HBV mutations in the core promotor region leading to a minimal viral replication, or mutations in the HBsAg-encoding region might explain the absence of serological recognition. Finally, it is possible that in some cases host immune mechanisms can maintain HBV infection in a latent state until transmission to another individual who subsequently develops a more active infection especially when immunosuppressive therapy is employed. Existence of HBsAg(-) HBV infections should be taken into account by the use of sensitive PCR tests for prevention of viral transmission in the settings of blood donations and organ transplants.     

Evaluation of hepatitis B virus replication and proteomic analysis of HepG2.2.15 cell line after cyclosporine A treatment

Aim： The effect of cyclosporine A （CsA） on hepatitis B virus （HBV） replication was investigated, and proteomics expression differentiation after CsA treatment was studied in order to provide clues to explore the effect of CsA on HBV replication. Methods： Methyl thiazolyl tetrazolium （MTT） assay was used to evaluate the cytotoxicity of CsA. The HBV replication level in the HBV genomic DNA transfected HepG2.2.15 cell line was determined by an ELISA analysis of hepatitis B surface antigens （HBsAg） and Hepatitis B e antigens （HBeAg） in culture supernatant, while the intracellular HBV DNA replication level was analyzed by slot blot hybridization. Two-dimensional electrophoresis was used to investigate the alteration of protein expression in HepG2.2.15 after CsA treatment in vitro. The differentially-expressed proteins were identified by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry combined with an online database search. Results： CsA was able to inhibit the expression of HBsAg, HBeAg, and HBV DNA replication in vitro in a dose-dependent manner. A proteomics analysis indicated that the expression of 17 proteins changed significantly in the CsA treatment group compared to the control group. Eleven of the 17 proteins were identified, including the overexpression of eukaryotic translation initiation factors （eIF） 3k, otubain 1, 14.3.3 protein, eIF2-1α, eIF5A, and the tyrosine 3/tryptophan 5-mono-oxygenase activation protein in CsA-treated HepG2.2.15 cells. The downregulation of the ferritin light subunit, erythrocyte cytosolic protein of 51 kDa （ECP-51）, stathmin 1/oncoprotein, adenine phosphoribosyl-transferase, and the position of a tumor protein, translationally- controlled 1, was shifted, suggesting it had undergone posttranslational modifications. Conclusion： Our study identified the inhibitory effect of CsA on HBV replication, and found that a group of proteins may be responsible for this inhibitory effect.     

Solution conformation of an immunodominant epitope in the hepatitis B virus preS2 surface antigen

We have determined the solution conformation of the major B cell epitope (residues 123-145, adrl23 hereafter) in the preS2 region of hepatitis B virus known to be associated with infection neutralization. The adrl23 shows an "L" shaped helix-turn-helix topology with two beta-turns formed by residues Ala(130)-Asp(133) and Asp(133)-Val(136) intervening the N- and C-terminal helices. The N-terminal alpha-helix consists of residues Ser(124)-Gln(129) whereas the C-terminal 3(10) helix is formed by residues Val(136)-Tyr(140). The beta-turns overlap partially with the putative "conformational" epitope. The overall topology of adrl23 is primarily maintained by hydrophobic interactions involving Phe(127), Leu(131), Leu(132), Val(136), and Tyr(140) that are clustered on one side of the molecule. An additional hydrophobic stabilization comes from Phe(141) that is buried inside the concave side of the molecule. A network of hydrogen bonds formed among Thr(125), His(128), and Arg(137) further contribute to the "boomerang-shaped" architecture of adrl23. The N-terminus of adrl23 is immobile due to a hydrogen bond between the N-terminal amide proton of Asn(123) and the hydroxyl oxygen of Thr(126). The side chains of Asp(133), Arg(135), Val(136), Leu(139), and Tyr(140) that were shown to be important for binding to a monoclonal antibody H8 mAb are surface exposed.     

Inhibition of hepatitis B virus by D-fraction from Grifola frondosa: synergistic effect of combination with interferon-alpha in HepG2 2.2.15

In this study, D-fraction extracted from Grifola frondosa (GF-D) and its combination with human interferon alpha-2b (IFN) were investigated for the inhibitory effect on hepatitis B virus (HBV) in HepG2 2.2.15 cells (2.2.15 cells). HBV DNA and viral antigens were analyzed by a quantitative real-time polymerase chain reaction and end-point titration in radioimmunoassays, respectively. The results showed that GF-D or IFN alone could inhibit HBV DNA in 2.2.15 cells with the 50% inhibitory concentration (IC50) of 0.59 mg/ml and 1399 IU/ml, respectively. We further investigated the combination of GF-D and IFN for anti-HBV activity and found that they synergistically inhibited HBV replication in 2.2.15 cells. In combination with 0.45 mg/ml GF-D, the apparent IC50 value for IFN was 154 IU/ml. This 9-fold increase in antiviral activity of IFN suggested that GF-D could synergize with IFN. These results indicate that GF-D, in combination with IFN, might provide a potentially effective therapy against chronic HBV infections.     

Serum hepatitis B surface antigen levels in the natural history of chronic hepatitis B infection

BACKGROUND The production of hepatitis B surface antigen (HBsAg) may evolve during long-lasting virus-host interactions in chronic hepatitis B (CHB). The impact of age on HBsAg production remains unclear. AIM To determine the age-specific distribution patterns of HBsAg and related factors during the natural course of CHB infection. METHODS Seven hundred and sixty-eight untreated HBsAg carriers were enrolled in the study. The parameters and distribution patterns of HBsAg were evaluated in relation to age and immune phases. RESULTS The HBsAg levels were significantly lower in the HBeAg-negative stage, with the lowest levels in inactive carriers. The HBsAg tended to decrease from hepatitis to cirrhosis and to hepatocellular carcinoma, and from Child-Pugh class A to B and to C. Age and HBV DNA were independently associated with HBsAg levels. In HBeAg-positive patients, the HBsAg levels were distributed in a triphasic-like decline pattern by 2 logs across age strata. For HBeAg-negative patients, the titres in inactive carriers exhibited a 2-log reduction, but remained unchanged over age strata in patients with HBeAg-negative hepatitis. The ratios of HBsAg/HBV-DNA were highest, but steadily decreased with age in inactive carriers, whereas the levels remained largely unchanged over the entire age strata in patients with HBeAg-negative hepatitis. CONCLUSIONS Age and HBV DNA levels are independent parameters of HBsAg levels. During the natural course of CHB infection, HBsAg levels decrease with age and disease progression, but the patterns are significantly different between the immune phases of CHB. This information may contribute to our understanding of the immunopathogenesis of chronic hepatitis B and management involving HBsAg quantification.