TRANSFORMING GENES IN DUCK HEPATIC CARCINOMA——mht(raf) AND Ha-ras

The transfection of NIH 3T3 cells was performed with DNAs from 2 duck primary hepatic carcinomas (DHC 40K, 9K) and I tumor-adjacent liver tissue (TAL, 9N). Transfectants were found from 40K, 9K and 9N DNAs. The secondary transfectants were obtained after transfection of RAT-1 cells with DNAs from primary transfectants. After hybridization with Ha-ras, Ki-ras, N-ras and mht oncogenes, it was found that duck mht (5.2 and 3.2 kb EcoRI fragments) and duck Ha-ras (3.4 kb EcoRI fragments) were present in all these transformants.This is the first report on transforming genes in duck primary hepatic cancer as well as tumoradjacent liver tissue     

ELISA法和化学发光法对血清中HIV-1HIV-2抗体、梅毒抗体和丙肝抗体的检测意义

目的探讨分析ELLSA法(酶联免疫吸附法)和化学发光法对血清中HIV-1/HIV-2抗体、梅毒抗体和丙抗体的临床检测意义。方法将382份脐血作为研究分析对象,分别采用酶联免疫吸附和化学发光法对所选标本进行检测,包含丙肝抗体、梅毒抗体、HIV-1/HIV-2抗体检测。结果采用ELLSA检测后HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体阳性率分别为0.52%、0.79%、1.05%。采用化学发光法检测HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体3种检测的阳性率分别为0.79%、1.05%、1.31%。标准抗体品梯度稀释后再进行检测,将此3种抗体稀释程度为10 pg/m L,三组抗体采用化学发光法进行检测,其结果均为阳性,采用酶联免疫吸附方式进行检测的只有梅毒抗体呈阳性,其余两项均为阴性。结论化学发光法作为临床进行血清中HIV-1/HIV-2抗体、梅毒抗体、丙肝抗体检测的首选方案,此检测方式具有更高的灵敏性。     

硒卡拉胶囊治疗慢性乙型肝炎

用硒卡拉胶囊治疗了慢性乙型肝炎 49例。结果表明 ,硒卡拉胶囊对慢性乙肝病人乏力等症状有明显改善作用 ,其ALT和SB复常率分别为 95 9%和 96 8% ,优于对照 (P <0 0 5 ) ,说明补硒治疗慢性乙肝有一定的改善临床症状、降酶等作用。     

用温度敏感高分子为载体的激光光声免疫分析及其应用

将单克隆抗乙肝表面抗原体抗与温度敏感高分子聚Ｎ－异丙基丙烯酰胺连接，中液相中与抗原、酶标抗体反应，通过热沉淀实现与游离酶标体抗体的异相分离。高分子载休的引入能减少非特异性吸附，罗大相载体聚乙烯酶标板更有利于激光光声技术高灵敏度优势的。同时，采用０．５％的十二烷基硫酸钠水溶液作显色终止剂，易与Ｈｅ－Ｎｅ激光波长匹配，实现了对低值乙肝表面抗原阳性血清的检测。     

Size‐selective purification of hepatitis B virus‐like particle in flow‐through chromatography: Types of ion exchange adsorbent and grafted polymer architecture

Hepatitis B virus‐like particles expressed in Escherichia coli were purified using anion exchange adsorbents grafted with polymer poly(oligo(ethylene glycol) methacrylate) in flow‐through chromatography mode. The virus‐like particles were selectively excluded, while the relatively smaller sized host cell proteins were absorbed. The exclusion of virus‐like particles was governed by the accessibility of binding sites (the size of adsorbents and the charge of grafted dextran chains) as well as the architecture (branch‐chain length) of the grafted polymer. The branch‐chain length of grafted polymer was altered by changing the type of monomers used. The larger adsorbent (90 μm) had an approximately twofold increase in the flow‐through recovery, as compared to the smaller adsorbent (30 μm). Generally, polymer‐grafted adsorbents improved the exclusion of the virus‐like particles. Overall, the middle branch‐chain length polymer grafted on larger adsorbent showed optimal performance at 92% flow‐through recovery with a purification factor of 1.53. A comparative study between the adsorbent with dextran grafts and the polymer‐grafted adsorbent showed that a better exclusion of virus‐like particles was achieved with the absorbent grafted with inert polymer. The grafted polymer was also shown to reduce strong interaction between binding sites and virus‐like particles, which preserved the particles’ structure.     

The Properties of an HBV Surface Antigen Protein Carrying the Binding Site for the Receptor of Hepatocytes——Its Formation of Surface Antigen Particles and Secretion From Discrete Cell Lines

A human hepatitis B virus (HBV) gene, which encodes the major surface antigen protein(S protein) carrying the hepatocyte receptor-binding site, was constructed with site-directed mutagenesis and in vitro recombination. When expressed in monkey kidney cell line COS-M6, this gene product (S309 protein) formed surface antigen (HBsAg) particles and secreted from the cells. It was stable within the cells and in the culture medium and could be immunoprecipitated with antisera directed against plasma-derived HBsAg or synthetic preS1 polypeptide. Isopycnic CsCl gradient centrifugation showed that the density of S309 protein particles (1.25 g/ml) was slightly higher than that of S protein particles. The S309 protein was readily secretable from hepatoma cell lines, and the amount secreted was comparable to that of the S protein. By contrast, only about 10% of the S309 protein was secreted from COS-M6 cells, and its appearance in culture medium was delayed. The efficiency of the secretion of the S309 protein can b     

Nucleotide Sequence of a Cloned Human HBV Mutant (pDKHBV) in Duck Hepatoma of Qidong County

This is the first report to describe the presence of an HBV-like DNA sequence in two hepatoma and their tumor surrounding liver tissues, one precancerous and one non-malignant liver tissue of ducks collected from Qidong County of China. The HBV-like sequences were either in an episomal form of 3.2 kb or in an integrated form of various sizes, while the DHBV DNA sequences (3.0 kb) were either present or absent in these tissues and in different size pattern. Furthermore, there was no evidence of cross-hybridization between HBV-like DNA sequences in duck and DHBV DNA. A 3.2 kb HBV-like DNA sequence has been cloned from one duck hepatoma (40 K), designated as pDKHBV. The 3218 bp full-length nucleotide sequence of this clone has been determined, which had no apparent homology with Duck Hepatitis B Virus (DHBV) genome, but was highly homologous to human HBV adw_2 subtype (99.0%). The sequence was composed of four open reading frames for HBV gene Pre-S/S, X, C and P respectively. In addition to multiple sites     

In vivo hepatitis B virus-neutralizing activity of an anti-HBsAg humanized antibody in chimpanzees

Previously, we constructed a humanized antibody (HuS10) that binds to the common a antigenic determinant on the S protein of HBV. In this study, we evaluated its HBV-neutralizing activity in chimpanzees. A study chimpanzee was intravenously administered with a single dose of HuS10, followed by intravenous challenge with the adr subtype of HBV, while a control chimpanzee was only challenged with the virus. The result showed that the control chimpanzee was infected by the virus, and thus serum HBV surface antigen (HBsAg) became positive from the 14(th) to 20(th) week and actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 19(th) and 23(rd) week, respectively. However, in the case of the study chimpanzee, serum HBsAg became positive from the 34(th) to 37(th) week, while actively acquired serum anti-HBc and anti-HBs antibodies appeared from the 37(th) and 40(th) week, respectively, indicating that HuS10 neutralized the virus in vivo and thus delayed the HBV infection. This novel humanized antibody will be useful in the immunoprophylaxis of HBV infection.     

彩色多普勒超声联合Fibroscan评分系统在慢性乙肝纤维化诊断中的应用研究

本研究探讨了彩色多普勒超声联合Fibroscan评分系统在慢性乙肝纤维化诊断中的应用价值。选取行超声引导下肝穿刺活检的慢性乙肝患者300例,其中235例有肝纤维化(观察组)和65例无肝纤维化(对照组),统计分析了这两组患者和不同肝纤维化分期患者的彩色多普勒超声参数[门静脉主干内径(mPVD)、脾静脉内径(SVD)、门静脉血流量(mPBF)、门静脉主干平均血流速度(mPVm)、门静脉淤血指数(CI)、脾静脉平均血流速度(sVm)、脾静脉血流量(sVBF)]和肝脏硬度值(LSM)。结果显示,各参数联合诊断CHB肝纤维化的AUC最高,为0.876(P<0.05);SVD、mPVD、CI、LSM与肝纤维化分期呈正相关,mPVm与肝纤维化分期呈负相关(P<0.05);LSM诊断肝纤维化分期的AUC最大(P<0.05)。可见采用彩色多普勒超声与FibroScan定向瞬时弹性成像系统检测SVD、mPVD、CI、mPVm、LSM参数可诊断CHB肝纤维化,且各参数均与肝纤维化程度密切相关,有望成为评估肝纤维化及严重程度的无创性检测方法。     

Identification of Potential HCV Inhibitors Based on the Interaction of Epigallocatechin-3-Gallate with Viral Envelope Proteins

Hepatitis C is affecting millions of people around the globe annually, which leads to death in very high numbers. After many years of research, hepatitis C virus (HCV) remains a serious threat to the human population and needs proper management. The in silico approach in the drug discovery process is an efficient method in identifying inhibitors for various diseases. In our study, the interaction between Epigallocatechin-3-gallate, a component of green tea, and envelope glycoprotein E2 of HCV is evaluated. Epigallocatechin-3-gallate is the most promising polyphenol approved through cell culture analysis that can inhibit the entry of HCV. Therefore, various in silico techniques have been employed to find out other potential inhibitors that can behave as EGCG. Thus, the homology modelling of E2 protein was performed. The potential lead molecules were predicted using ligand-based as well as structure-based virtual screening methods. The compounds obtained were then screened through PyRx. The drugs obtained were ranked based on their binding affinities. Furthermore, the docking of the topmost drugs was performed by AutoDock Vina, while its 2D interactions were plotted in LigPlot+. The lead compound mms02387687 (2-[[5-[(4-ethylphenoxy) methyl]-4-prop-2-enyl-1,2,4-triazol-3-yl] sulfanyl]-N-[3(trifluoromethyl) phenyl] acetamide) was ranked on top, and we believe it can serve as a drug against HCV in the future, owing to experimental validation.