丹参注射液对抗结核药物所致肝损害的预防作用

动物实验及临床应用均证实丹参注射液对肝损害具有治疗及预防作用。我们自1993年以来，应用丹参注射液预防抗结核药物引起的肝损害，取得满意效果，报告如下。工临床资料1．亚病例选择60例病人为住院的活动型肺结核患者，男40例，女20例，平均年龄32岁（19～70岁）。其中力型5例，巨型36例，IV型11例，V型8例。甲、丙肝抗体阴性，血清ALT、TB、DB、A／G均正常。全部病例按入院顺序随机分为两组：治疗组35例，对照组25例。两组在性别、年龄、病程及分型上具有可比性。1．2治疗方法两组抗结核药物治疗均采用ZSHRZ／4HR方案，剂量为常…     

生物人工肝治疗肝衰竭的临床应用

临床上各种原因所致肝衰竭的治疗问题 ,一直是最为棘手、最需要尽快解决的问题。因肝脏具有复杂的代谢、生物合成、生物转化、解毒及排泄等功能 ,故肝衰竭将造成严重的代谢紊乱和毒物聚积 ,并常导致肝外其它脏器功能衰竭 ,形成多器官功能衰竭 (ＭＯＦ) ,病死率极高。尽管传统内科对症支持疗法有了长足进步 ,肝衰竭的病死率仍居高不下 ,即使在西方国家暴发性肝衰竭 (ＦＨＦ)病死率亦高达80 %左右 ,而ＦＨＦ中达Ⅳ期肝性脑病者病死率在 95 %以上[1] 。鉴于此 ,国外学者长期以来一直致力于肝衰竭非药物治疗方法的研究 ,主要措施为体外生物人工…     

人工冬眠在大面积烧伤合并精神病患者的应用

目的探讨人工冬眠在大面积烧伤合并精神病患者治疗中的应用。方法病人度过休克期后采用手术植皮及湿润包扎疗法,在人工冬眠的配合下防止并发症及营养支持治疗。结果2例治疗无并发症发生,1例家属放弃治疗。结论人工冬眠配合大面积烧伤合并精神病治疗效果好。     

艾滋病合并结核性脑膜炎治疗研究进展

结核病为艾滋病患者中最常见的机会性感染,因此,结核性脑膜炎在艾滋病患者中的发病率也较高。然而,结核性脑膜炎合并艾滋病的患者治疗较单纯结核性脑膜炎患者更为复杂。文章从抗结核治疗、辅助治疗以及抗反转录病毒治疗三个方面对艾滋病合并结核性脑膜炎患者的治疗进展进行综述。     

L02细胞建立人鼠嵌合肝动物模型

目的:研究L02细胞移植到具有正常免疫活性的大鼠肝内的存活情况.方法:SD大鼠出生前宫内腹腔注射正常人L02肝细胞,诱导胎鼠对人L02肝细胞产生免疫耐受,出生2wk时经脾移植DiI染色后的人L02肝细胞,建立人鼠嵌合肝动物模型.采用免疫荧光、SP免疫组化、DiI荧光示踪等方法,分别检测人白蛋白、特异性人增殖细胞核抗原PCNA(proliferatingcellnuclearantigen)以及在荧光显微镜下观察人L02肝细胞在鼠肝内的分布.结果:于移植后1,2,4,6,8,10wk在荧光显微镜下观察到人L02肝细胞在鼠肝内的动态分布;移植后2,4,6,8wk大鼠均检测出人白蛋白,4wk时分泌最多;移植后2,4,6wk检测出特异性人增殖细胞核抗原PCNA,以4wk发现PCNA阳性细胞最多.结论:移植的人L02肝细胞在具有正常免疫活性的免疫耐受鼠体内能够存活、增殖,并产生人白蛋白.     

蛋白转导结构域介导乙型肝炎病毒聚合酶末端蛋白重链可变区抗体体外抑制病毒的复制

Objective To study a functional variable fragment of heavy chain(VH)antibody against the terminal protein(TP)region of hepatitis B virus(HBV)polymerase introduced by human immunodeficiency virus Tat protein transduction domain(TAT)and the inhibitive activity of TAT-VH on the replication of HBV in vitro.Methods The gene encoding TAT-VH was cloned into prokaryotic expression vector pET28a(+).Recombinant plasmid was transduced into E coli BL21(DE3)LysS,then the protein was expressed and purified.The purified TAT-VH fusion protein was added into HepG2.2.15 cell culture.The transduction efficiency was evaluated by indirect fluorescence assay(IFA).The cytotoxicity of TAT-VH was detected by Methabenzthiazuron(MTT)assay.HBV DNA level in HepG2.2.15 cell culture was measured using quantitative polymerase chain reaction(PCR).The data were analyzed by one-factor analysis of variance and t test.Results TAT-VH fusion protein was successfully expressed and purified.It was confirmed by IFA and MTT assay that TAT-VH was introduced into HepG2.2.15 cells and the cell growth was not affected.The level of HBV DNA in supernatant of HeDG2.2.15 cell culture with 5 000 nmol/L TAT-VH was(1.211±0.132)lg copy/mL,which was significantly lower than control group[(5.325±0.041)lg copy/mL,t=72.91,P＜0.05].Meanwhile,the level of intracellular HBV DNA was(3.521±0.411)lg copy/mL,which was significantly lower than control group[(8.532±0.132)lg copy/mL.t=28.41,P＜0.05].Conclusion The HBV replication is inhibited by anti-TP TAT-VH antibodies in vitro,which provides valuable experimemal basis for developing therapy of HBV infection with intracellular antibody.     

蛋白转导结构域介导乙型肝炎病毒聚合酶末端蛋白重链可变区抗体体外抑制病毒的复制

Objective To study a functional variable fragment of heavy chain(VH)antibody against the terminal protein(TP)region of hepatitis B virus(HBV)polymerase introduced by human immunodeficiency virus Tat protein transduction domain(TAT)and the inhibitive activity of TAT-VH on the replication of HBV in vitro.Methods The gene encoding TAT-VH was cloned into prokaryotic expression vector pET28a(+).Recombinant plasmid was transduced into E coli BL21(DE3)LysS,then the protein was expressed and purified.The purified TAT-VH fusion protein was added into HepG2.2.15 cell culture.The transduction efficiency was evaluated by indirect fluorescence assay(IFA).The cytotoxicity of TAT-VH was detected by Methabenzthiazuron(MTT)assay.HBV DNA level in HepG2.2.15 cell culture was measured using quantitative polymerase chain reaction(PCR).The data were analyzed by one-factor analysis of variance and t test.Results TAT-VH fusion protein was successfully expressed and purified.It was confirmed by IFA and MTT assay that TAT-VH was introduced into HepG2.2.15 cells and the cell growth was not affected.The level of HBV DNA in supernatant of HeDG2.2.15 cell culture with 5 000 nmol/L TAT-VH was(1.211±0.132)lg copy/mL,which was significantly lower than control group[(5.325±0.041)lg copy/mL,t=72.91,P＜0.05].Meanwhile,the level of intracellular HBV DNA was(3.521±0.411)lg copy/mL,which was significantly lower than control group[(8.532±0.132)lg copy/mL.t=28.41,P＜0.05].Conclusion The HBV replication is inhibited by anti-TP TAT-VH antibodies in vitro,which provides valuable experimemal basis for developing therapy of HBV infection with intracellular antibody.     

肝干细胞研究方法进展

当前 ,国外有关肝干细胞 (又称肝祖细胞或肝干 /祖细胞 )研究进展迅速 ,但国内研究甚少 ,主要原因是方法学受限。肝干细胞方法学较为复杂 ,涉及其活化、分离培养、细胞特异性标志物筛选及细胞体内演变的鉴定等。现将有关研究方法进展综述如下。一、分离培养方法 (图 1 )肝干细胞的分离与培养是研究其生物学特性以及通过细胞移植治疗肝病的必要前提 ,因而倍受重视。Ｙａｓｕｉ等[1 ] 从ＬＥＣ(Ｌｏｎｇ ＥｖａｎｓＣｉｎｎａｍｏｎ)大鼠分离肝脏细胞 ,继用胶原酶、蛋白酶Ｅ、胰蛋白酶及ＤＮＡ酶破坏大肝细胞 ,然后将细胞悬液覆盖于 1 .0 35～…     

晚期艾滋病患者使用联合艾博韦泰ART方案效果

目的 了解联合使用艾博韦泰（ABT）的ART方案用于晚期艾滋病患者的安全性和有效性。方法 回顾性收集联合使用ABT治疗的晚期艾滋病患者资料,分析临床特征,评估该方案的安全性、有效性,观察后续更换为不含ABT的方案后抗病毒疗效。结果 收集182例联合使用ABT进行ART的艾滋病患者,156例（85.7%）合并机会性感染或恶性肿瘤;126例（69.2%）合并脏器功能损伤或慢性基础疾病。平均使用ABT(35.2±9.7)天HIV RNA(2.5log10 copies/mL)较基线显著降低（Z=-7.898,P=0.000）,下降了3.1 log10 copies/mL,CD4细胞（113个/μL）较基线显著升高（Z=-4.650,P=0.000）,增长了57个/μL;用药期间未发生ABT相关不良事件。停用ABT后60.2%(50/83)转换为整合酶抑制剂为核心的三药方案,37.3%(31/83)转为含NNRTI的方案,治疗1年73.5%的患者HIV RNA<50 copies/mL,CD4细胞（157.0个/μL）较停用ABT时显著升高（Z=-1.947,P=0.049）;治疗2年8...     

肝移植后肝炎病毒再感染的防治进展

人体肝移植是目前各种终末期肝病及肝功能衰竭最有效的治疗手段。肝炎病毒特别是乙型肝炎病毒 (ＨＢＶ)与丙型肝炎病毒 (ＨＣＶ)所致的急慢性严重肝病是肝移植的重要适应证 ,但此类病人肝移植后常发生肝炎病毒的再感染或复发感染 ,再感染可导致植入肝脏发生炎症、纤维化及功能衰竭 ,严重影响移植肝存活率及病人生存率 ,是影响预后的关键因素。近年来国外在肝移植后ＨＢＶ及ＨＣＶ再感染防治方面积累了一些成功的经验 ,现就有关文献综述如下。一、 再感染的发病机制肝移植后发生肝炎病毒再感染的机制较为复杂 ,许多环节尚待进一步阐明。由于…