酒精性肝病实验室检查及远期预后观察

病理检查诊断酒精性脂肪肝（ＡＦ）１０例，酒精性肝炎（ＡＨ）１５例，酒精性肝硬化（ＡＣ）１５例，轻微病变（ＭＣ）５例，与病理及ＨＢＶ－Ｍ诊断ＣＡＨ１０例比较。ＧＧＴ试验显示ＡＦ平均２４０±５２．５μｍｏｌ／ＬＡＨ３８１±１８．４μｍｏｌ／Ｌ。（最高１６００μｍｏｌ／Ｌ），ＡＣ２０５±６０．５μｍｏｌ／Ｌ，ＣＡＨ７６．９±２０．７μｍｏｌ／Ｌ。ＡＦ、ＡＨ与ＣＡＨ比较差异显著（Ｐ＜０．０１）。ＡＦ、ＡＨ戒酒后，ＧＧＴ恢复好转，再酗明显上升，结合嗜酒史，有诊断参考意义。甘氨胆酸试验（ＲＩＡ）示ＡＣ增高显著（３２５８±５０．２ｍｇ／Ｌ，最高７０００ｇ／Ｌ，与ＡＨ（１５３５±ｌ９．２ｍｇ／Ｌ）比较（Ｐ＜０．０１），对判断预后有参考价值。ＡＨ病与ＡＨ２例较西方国家轻，ＡＣ年龄接近（４８．１１±１４．２岁）。共随访１～１２年（平均６．３年）。戒酒后ＡＦ、ＭＣ完全恢复，ＡＣ于继续酗酒后２－４年内死于肝衰竭，ＥＶＢ与ＨＣＣ死亡。戒酒为终身治疗，支持治疗辅之，而治疗乙醇依赖及戒酒综合征（ＡＷＳ）乃达到戒酒的关键。     

老年不稳定型心绞痛患者纤维蛋白、纤维蛋白原及其降解产物的变化

目的探讨凝血系统在老年人不稳定型心绞痛（ＵＡ）发生和发展中的作用。方法采用一组抗纤维蛋白单克隆抗体（ＳＺ－５８、６４、６５）酶免疫分析法测定２６例老年ＵＡ患者、２４例稳定型心绞痛（ＳＡ）患者和２０例健康老年人（对照组）血浆纤维蛋白原（Ｆｇ）、可溶性纤维蛋白复合物（ＳＦＣ）及血清纤维蛋白降解产物（ＦＤＰ）的浓度。结果ＵＡ患者心绞痛发作时血浆Ｆｇ、ＳＦＣ及血清ＦＤＰ浓度（分别为３．７±０．６ｇ／Ｌ、４９．６±１９．３ｍｇ／Ｌ、３２５．６±７９．４μｇ／Ｌ）显著高于ＳＡ患者心绞痛发作时（分别为３．２±０．６ｇ／Ｌ、２０．９±１０．４ｍｇ／Ｌ、２２４．４±４７．４μｇ／Ｌ）和健康对照组（均为Ｐ＜０．０１），后两者间差异无显著性。ＵＡ发作终止后血浆Ｆｇ和血清ＦＤＰ浓度高于健康对照组，但差异无显著性。结论血栓形成是老年人ＵＡ发生和发展的重要因素之一。     

咳嗽变异型哮喘患者11例咳嗽受体敏感性观察

目的　探讨咳嗽变异型哮喘（ＣＶＡ）的咳嗽机制。方法　对１１ 例咳嗽变异型哮喘患者、７ 例哮喘稳定期患者、１０ 例急性支气管炎迁延期患者和１０ 例健康成年人,以组胺吸入进行激发感应ＦＥＶ１ 下降２０ ％ 的浓度（ＰＣ２０ＦＥＶ１）和吸入酒石酸测定咳嗽受体敏感性（ＣＲＳ５）。结果　ＣＶＡ患者、哮喘稳定期患者、急性支气管炎迁延期患者和健康成年人的ＰＣ２０ＦＥＶ１ 分别为（３－７５ ±１－５８）ｇ／Ｌ、（２－０１ ±１－４６）ｇ／Ｌ、（１７－４ ±１－５２）ｇ／Ｌ 和（１８－０３ ±１－６９）ｇ／Ｌ,ＣＶＡ 患者显著降低（ Ｐ ＜ ０－０１）;ＣＲＳ５ 分别为（３８－０１ ±２－０４）ｇ／Ｌ、（８２－２８ ±１－４５）ｇ／Ｌ、（３５－８８ ±２－１８）ｇ／Ｌ、（１１４－８１±１－３３）ｇ／Ｌ,ＣＶＡ 患者显著降低（ Ｐ＜ ０－０１） 。结论　ＣＶＡ 患者咳嗽受体敏感性的阈值明显降低。     

原发性肝癌患者血清甲状腺激素变化

目的观察原发性肝癌（ＰＨＣ）患者血清甲状腺激素的变化。方法应用放射免疫法（ＲＩＡ）检测２４例ＰＨＣ患者的血清Ｔ３，Ｔ４，ＴＳＨ和ｒＴ３，并以２４例健康成人作对照。结果ＰＨＣ组血清Ｔ３为１．６０±０．１４ｎｍｏｌ／Ｌ，而对照组为２．３７±０．０８ｎｍｏｌ／Ｌ（Ｐ＜０．００１）。ＰＨＣ切除组血清Ｔ３，Ｔ４和ｒＴ３分别为１．９４±０．１６ｎｍｏｌ／Ｌ，１３４．３４±１１．４９ｎｍｏｌ／Ｌ和２．６１±０．３９；而ＰＣＨ未切除组分别为１．２２±０．１７ｎｍｏｌ／Ｌ（Ｐ＜０．０５），１０４．０１±７．２４ｎｍｏｌ／Ｌ（Ｐ＜０．０５）和１．３５±０．３６（Ｐ＜０．０５）。结论ＰＨＣ患者血清Ｔ３降低，晚期患者出现血清Ｔ４降低和Ｔ３／ｒＴ３比值降低。     

环孢素A治疗难治性肾病综合征

目的研究环孢素Ａ（ＣｓＡ）治疗难治性肾病综合征的临床疗效并探讨其机制。方法对３０例肾上腺皮质激素依赖或抵抗的肾病综合征（ＮＳ）患者联合使用了（ＣｓＡ，５ｍｇ·ｋｇ－１·ｄ－１）和泼尼松（３０ｍｇ／ｄ）治疗，并测定了ＣｓＡ治疗前后病人的血生化指标、免疫指标及２４小时尿蛋白排出量。结果２４小时尿蛋白排出量由治疗前的平均７．６７±３．００ｇ／２４ｈ降至治疗后第１、２、３个月的４．９３±３．６４ｇ／２４ｈ、３．５２±２．９４ｇ／２４ｈ、２．２３±１．６０ｇ／２４ｈ；血浆白蛋白由治疗前２７．３±６．４ｇ／Ｌ升至３０．９±８．８ｇ／Ｌ、３４．３±７．６ｇ／Ｌ、３６．０±７．２ｇ／Ｌ；治疗前后血肌酐、ＢＵＮ、尿酸、ＣＤ４／ＣＤ８和可溶性白细胞介素２（ＩＬ２）受体无明显变化，而治疗２个月后外周血单个核细胞产生血清ＩＬ２显著下降。治疗后激素抵抗组ＮＳ完全缓解、部分缓解和无效分别为２２．７％，５０．０％和２７．３％；在激素依赖组分别为５７．１％，４２９％和０。有５例患者因肝肾功能损害或严重腹泻而停用ＣｓＡ。结论本研究提示ＣｓＡ联合小剂量皮质激素治疗难治性肾病综合征，尤其是激素依赖的患者是安全和有效的。     

抗HCV阳性慢性肝病患者HBV感染及前C／C区基因变异

探讨ＨＢＶ基因变异在ＨＢｓＡｇ阴性抗ＨＣＶ阳性慢性肝病中的意义。方法用巢式聚合酶链反应（ＰＣＲ）与限制片段长度多态性相结合，对５６例慢性肝病患者６例ＨＢｓＡｇ阴性抗ＨＣＶ阳性（Ａ组）．１９例ＨＢｓＡｇ阳性抗ＨＣＶ阳性（Ｂ组）及３１例单独ＨＢＶ成染（Ｃ组）进行前Ｃ区密码２８终止变异（Ａ８３）和Ｃ区密码９７异亮氨酸亮氨酸变异（Ｌ９７）分析。结果Ａ组和Ｂ组Ａ８３和Ｌ９７变异检出率分别为３３％和４２％显著低于Ｃ组８１％（Ｐ值＜０．００１）：而Ａ组和Ｂ组间无统计学差异（Ｐ值＞０．０５）。结论ＨＢＶ和ＨＣＶ双重感染ＨＢｓＡｇ阴性与Ａ８３和Ｌ９７变异无相关。     

拉米夫定、治肝灵冲剂联合治疗慢性乙型肝炎近期疗效研究

研究单用拉米夫定（３－ＴＣ）及联合应用中药治肝灵冲剂对慢性乙型肝炎（ＣＨＢ）的治疗作用，探讨两者联合治疗的近期疗效和安全性。５０例ＣＨＢ患者随机分为Ａ，Ｂ两组，Ａ组采用３－ＴＣ和治肝灵冲剂联合治疗；Ｂ组单用３—ＴＣ，疗程均为６个月，治疗前、治疗２周、４周、８周、１２周、１６周，２０周、２６周分别检测ＨＢＶ标志、肝功能，血清学指标，肾功能，并记录治疗期间临床和实验室检查发生的一切不良事件。治疗２６周后Ａ组血清ＨＢｅＡｇ转阴率６４．０％（１６／２５）．明显高于Ｂ组１６．０％（４／２５）Ｐ＜０．０１． Ａ组与Ｂ组分别有４例（１６．０％），２例（８．０％）实现ＨＢｅＡｇ血清转换。Ｐ＞０．０５。转阴患者未发现ＨＢＶ前Ｃ区变异株。Ａ组和Ｂ组ＨＢＶＤＮＡ转阴例数分别为２４（９６．０％）和２３（９２．０％）Ｐ＞０．０５。治疗结束时Ａ组和Ｂ组ＡＬＴ的复常率分别为８８．０％（２２／２５）、６４．０％（１６／２５），Ｐ＜０．０５。采用３－ＴＣ与治肝灵冲剂联合治疗ＣＨＢ，短期疗效明显优于单一用药组，且副作用少，是安全、有效的治疗ＣＨＢ的方案。     

慢性肝炎患者高压氧治疗前后肝组织中HBsAg、HBcAg检测初步探讨

目的　观察慢性肝炎（ＣＨ） 高压氧（ ＨＢＯ） 治疗前后患者的免疫功能、肝组织中ＨＢｓＡｇ 、ＨＢｃＡｇ 变化。方法：用纯氧单仓治疗（２－５ＭＰａ ,２ｈ／ｄ ,１０ｄ／ｃｙｃ ,６ｃｙｃ）３０ 例ＣＨ,治疗前后用美国ＢＥＣＭＡＮ 公司生产ＡｒｒａｙＴＭＰｒｏｔｅｉｎ Ｓｙｓｔｅｍ ３６０ 测全部患者静脉血白蛋白（ Ａ） 、γ球蛋白（γ） 、ｌｇＡ、ｌｇＧ、ｌｇＭ、和补体Ｃ３ 、Ｃ４ ;二次肝穿刺活检,ＡＢＣ 法检测肝组织中ＨＢｓＡｇ 、ＨＢｃＡｇ 。取算术均数行ｔ 检验。结果：ＨＢＯ 治疗后静脉血γ、ｌｇＧ、ｌｇＭ、明显降低,Ａ、Ｃ３ 明显升高,治疗前后差异显著（ Ｐ＜０－０５） ;肝组织中ＨＢｓＡｇ 、ＨＢｃＡｇ 变化不明显,治疗前后差异不显著（ Ｐ＞ ０－０５） 。结论：ＨＢＯ 治疗ＣＨ,可抑制患者的免疫功能,对ＨＢｓＡｇ 、ＨＢｃＡｇ 抑制作用不明显     

HClO3,NaCl和H2SO4在温和酸消化砷铈催化反应测定尿碘的影响

研究了在温和酸消化法砷－铈催化反应体系中ＨＣｌＯ３、ＮａＣＩ和Ｈ２ＳＯ４的作用和影响。采用尿碘测定标准方法［５］对ＨＣｌＯ３、ＮａＣｌ和Ｈ２ＳＯ４在不同浓度下进行对比实验，结果表明ＨＣｌＯ３、ＮａＣｌ对砷—铈反应均具有催化作用，Ｈ２ＳＯ４能增大Ｃｅ４＋的吸光度，当溶液中Ｈ２ＳＯ４的浓度增大到０．４ｍｏｌ／Ｌ及以上时，对于１×１０－３ｍｏｌ／ＬＣｅ４＋，其光密度趋于恒定。Ｈ２ＳＯ４加快ＨＣｌＯ３与Ａｓ２Ｏ３－ＮａＣｌ试液的氧化还原反应速度，当Ｈ２ＳＯ４的浓度增大到０．８ｍｏｌ／Ｌ时，该反应明显发生，当Ｈ２ＳＯ４的浓度增大到１．０ｍｏｌ／Ｌ及以上时，该反应剧烈发生，并伴有大量Ｃｌ２和ＣｌＯ２逸出。指出在温和酸消化法测定尿碘的反应体系中，ＮａＣｌ的浓度以２０ｇ／Ｌ，Ｈ２ＳＯ４的浓度以０．５ｍｏｌ／Ｌ为适宜。     

胃癌组织5q微卫星不稳定性与APC/MCC基因杂合性缺失的研究

目的探讨５ｑ微卫星不稳定性（ＭＳＩ）与ＡＰＣ／ＭＣＣ基因杂合缺失（ＬＯＨ）的关系。方法应用ＰＣＲ－ＳＳＬＰ及ＰＣＲ－ＲＦＬＰ技术分析５２例手术切除胃癌组织中ＭＳＩ及ＡＰＣ／ＭＭＣ基因ＬＯＨ。结果５ｑＭＳＩ检出率为３４．０％（１６／４７）,ＡＰＣ／ＭＣＣ基因ＬＯＨ率为３１．４％（１１／３５）。早期胃癌５ｑＭＳＩ阳性率为６６．７％（２／３）,ＡＰＣ／ＭＣＣＬＯＨ率为５０％（１／２）;进展期分别为３１．８（１４／４４）,３０．３％（１０／３３）。两组间无显著差别（Ｐ＞０．０５）。ＭＳＩ及杂合缺失与肿瘤大小、浸润深度、淋巴结转移及临床分期无关。粘液（印戒）细胞癌ＡＰＣ／ＭＣＣＬＯＨ率（５５．６％）显著高于高、中分化管状腺癌（Ｐ＜０．０５）。胃、肠两型胃癌５ｑＭＳＩ及ＡＰＣ／ＭＣＣＬＯＨ差异无显著性及５ｑＭＳＩ与ＡＰＣ／ＭＣＣＬＯＨ无相关性（Ｐ＞０．０５）。结论染色体５ｑＭＳＩ有ＡＰＣ／ＭＣＣ基因ＬＯＨ在两型胃癌的早期发生及发展中起一定作用。染色体５ｑ可能是胃癌的易感部位。     

Cleavage of C2 by C1s into the antigenically distinct fragments C2a and C2b: demonstration of binding of C2b to C4b.

The activation of complement component C2 by C1s is a major reaction step leading to the assembly of two related macromolecular enzymes in the classical complement pathway C3 convertase and C5 convertase. The present studies clearly document the smaller fragment, C2b, that results when human C2 reacts with C1s. We have identified and characterized C2b (34,000 daltons) as a single protein on disc electrophoresis and immunoelectrophoresis. C2a (73,000 daltons), the larger fragment from this reaction, has a more acidic nature and C2b is more basic. These fragments can also be detected by their different antigenic determinants. When the C2-C4b complex is activated in the fluid phase by C1s and allowed to decay, it dissociates into C2a and the C2b-C4b complex. Furthermore, when C2 is bound to C4b-Sepharose and then reacted with C1s, only the C2a fragment is released from the solid phase C2-C4b-Sepharose into the fluid phase, and the C2b fragment remains noncovalently bound to C4b-Sepharose. These results suggest that the C2b portion of C2 contains a stable binding site for C4b and, after the decay release of C2a from this C3 convertase, the C2b fragment remains bound. Thus, the decay release of C2a may represent a temperature-dependent dissociation from C2b.     

Penetrance of the C28Y/C282Y genotype of the HFE gene

OBJECTIVE: Hereditary hemochromatosis is a common genetic disease caused by accumulation of iron in the body. Most cases are homozygous for the C282Y mutation in the HFE gene, but only a minority of homozygotes will ever suffer from clinical hemochromatosis. Estimates of the penetrance of the C282Y/C282Y genotype vary greatly. The purpose of this study was to estimate the penetrance using a stringent definition, i.e. liver cirrhosis. MATERIAL AND METHODS: The results from previous phenotypic population screening for hereditary hemochromatosis were combined with findings in hospital databases in order to estimate the number of C282Y homozygotes with and without liver cirrhosis in a Norwegian county. The penetrance of the C282Y/C282Y genotype was estimated as the fraction of C282Y homozygotes with liver cirrhosis. We also calculated the expected number of male C282Y homozygotes with liver cirrhosis using figures for age-specific accumulated risk. RESULTS: The prevalence of liver cirrhosis in male homozygotes is between 3.4% and 5.0%. This figure is compatible with an accumulated risk of liver cirrhosis that increases from 0.2% at 35 years to about 10% at 65 years of age. In female homozygotes, the prevalence of liver cirrhosis is 0.3%. CONCLUSIONS: A small but significant number of Norwegian male C282Y homozygotes will contract liver cirrhosis if their hemochromatosis is not diagnosed and treated in time. The penetrance is much lower in women than in men.     

Mutants of complement component C3 cleaved by the C4-specific C1-s protease.

To identify some of the structural features determining specific protease recognition of complement components C3 and C4, we used site-specific mutagenesis to construct mutants of murine C3 that are cleaved by the C4-specific C1-s protease. Insertion of three amino acid residues corresponding to residues at the C1-s cleavage site of human C4 into murine C3 at the analogous C3 convertase cleavage site was adequate to render the mutant protein susceptible to C1-s cleavage. In addition, insertion of C3-specific residues at the same site or introduction of the C4-specific residues as substitutions rather than as an insertion also rendered the site susceptible to cleavage, but with 10- to 50-fold lower efficiencies, and insertion of even a single amino acid residue affected recognition by C1-s. Finally, insertion of amino acid residues into mC3 partially inhibited cleavage by the alternative-pathway C3 convertase, with insertion of C3- or C4-specific residues giving about the same level of inhibition. A simple interpretation of these data is that C1-s cleavage is dependent primarily on steric accessibility and on recognition of specific amino acid residues at the cleavage site, whereas C3 convertase cleavage is dependent primarily on specific interactions distal to the cleavage site, with only relatively weak, non-C3-specific interactions at the cleavage site itself.     

Activated protein C induces the release of microparticle-associated endothelial protein C receptor

Activated protein C (APC) treatment is now used for patients with severe sepsis. We investigated its effect in vitro on primary, physiologically relevant cells and demonstrate a novel mechanism of endothelial protein C receptor (EPCR) release that is not inhibited by metalloproteinase inhibitors. Exposure of human umbilical vein endothelial cells or monocytes to APC (6.25-100 nM) results in the release of EPCR-containing microparticles, as demonstrated by confocal microscopy and characterized through flow cytometry, enzyme-linked immunosorbent assay quantitation of isolated microparticles, and Western blotting. The phenomenon is time- and concentration-dependent and requires the APC active site, EPCR, and protease activated receptor 1 (PAR1) on endothelial cells. Neither protein C nor boiled or D-Phe-Pro-Arg-chloromethylketone-blocked APC can induce microparticle formation and antibody blockade of EPCR or PAR1 cleavage and activation abrogates this APC action. Coincubation with hirudin does not alter the APC effect. The released microparticle bound is full-length EPCR (49 kDa) and APC retains factor V-inactivating activity. Although tumor necrosis factor-alpha (10 ng/mL) can also induce microparticle-associated EPCR release to a similar extent as APC (100 nM), it is only APC-induced microparticles that contain bound APC. This novel observation could provide new insights into the consequences of APC therapy in the septic patient.     

Similarity between phospholipase C and the regulatory domain of protein kinase C

We report that a 100 residue segment in the carboxy-terminus half of phosphatidylinositol-specific phospholipase C is similar to a segment in the amino-terminus half of protein kinase C. The computer-based comparison score is 9.5 standard deviations higher than that of 2500 comparisons of randomized sequences of these segments (P = 10(-21), suggesting that the two segments have similar biological properties. Phospholipase C has a segment that is homologous to part of the non-catalytic domain of src kinase and other tyrosine protein kinases. The similarity of phospholipase C with protein kinase C, a serine/threonine kinase suggests that novel exon shuffling occurred among serine/threonine and tyrosine kinases and phospholipase C.     

Epidemiology of the Hepatitis C Virus

According to WHO estimations, about 3 % of the world population may be infected with the hepatitis C virus. The relative prevalences of subtypes of this virus vary in different geographic areas. The main known routes of transmission are parenteral; intravenous drug abuse, contaminated injection devices and receipt of unscreened blood. Sexual, vertical, household and nosocomial transmissions may occur, but seem to be rare. The risk of screened blood or blood products is now almost eliminated, but unscreened blood is a considerable risk in areas where screening is economically not possible. The future impact of this virus is greatly dependent on the trends in intravenous drug use as well as the possible emergence of increased late morbidity among present asymptomatic carriers during the next decades.     

Quantitation of the Human Component C4: Definition of C4 Q0 Alleles and C4A Duplications

We determined the C4 plasma concentrations of 48 genotypically CA4- and C4B-defined unrelated individuals from 34 families with a total of 196 members by an enzyme-linked immunosorbent assay using Rg:1,2 (C4A) and Ch:1 (C4B) specific monoclonal antibodies. The results obtained allowed the establishment of rules for the detection of C4 Q0 alleles in the heterozygous form and of C4A gene duplications. In the present study seven homoduplications of the C4A 3 allotype were defined which had not been detected by allotyping. This procedure allows the simple, reliable, and quick determination of Rg:1,2 and Ch:1 plasma levels which are not influenced by daily rhythms of C4 production.     

Expression of recombinant human complement C1q allows identification of the C1r/C1s-binding sites

Complement C1q is a hexameric molecule assembled from 18 polypeptide chains of three different types encoded by three genes. This versatile recognition protein senses a wide variety of immune and nonimmune ligands, including pathogens and altered self components, and triggers the classical complement pathway through activation of its associated proteases C1r and C1s. We report a method for expression of recombinant full-length human C1q involving stable transfection of HEK 293-F mammalian cells and fusion of an affinity tag to the C-terminal end of the C chain. The resulting recombinant (r) C1q molecule is similar to serum C1q as judged from biochemical and structural analyses and exhibits the characteristic shape of a bunch of flowers. Analysis of its interaction properties by surface plasmon resonance shows that rC1q retains the ability of serum C1q to associate with the C1s-C1r-C1r-C1s tetramer, to recognize physiological C1q ligands such as IgG and pentraxin 3, and to trigger C1r and C1s activation. Functional analysis of rC1q variants carrying mutations of LysA59, LysB61, and/or LysC58, in the collagen-like stems, demonstrates that LysB61 and LysC58 each play a key role in the interaction with C1s-C1r-C1r-C1s, with LysA59 being involved to a lesser degree. We propose that LysB61 and LysC58 both form salt bridges with outer acidic Ca²⁺ ligands of the C1r and C1s CUB (complement C1r/C1s, Uegf, bone morphogenetic protein) domains. The expression method reported here opens the way for deciphering the molecular basis of the unusual binding versatility of C1q by mapping the residues involved in the sensing of its targets and the binding of its receptors.     

Human protein C and activated protein C Components of the human anticoagulation system

Protein C, the zymogen form of the anticoagulant protein, activated protein C, is a member of the vitamin K-dependent class of proteins that function in generation and control of formation of blood clots. This plasma protein consists of a series of domain regions that are qualitatively similar to those present in human coagulation factors VII, IX, and X, and that appear to be related to specific properties of these proteins. With the recent advent of rigorous application of genetic engineering strategies to this system, and the continuing discoveries and characterization of genotypes and phenotypes of protein C in patients, great progress has been made in understanding structure-function relationships of protein C and activated protein C. This review is a summary and synthesis of recent pertinent studies with an emphasis on these topics.