人白细胞干扰素治疗带状疱疹疗效观察

近年国外有用核苷类药物和干扰素治疗带状疱疹取得一定效果。我们对连续收治的33例病人应用干扰素治疗取得好的疗效,现报告如下:     

病毒感染与病毒受体

一、引言受体这一概念最早是由著名化疗学家Ehrlich在1909年提出的。他认为受体是一些特殊的化学基因,能与某些药物发生结合,并初步提出了“锁与钥匙,(Key and Lock)学说。继后,许多学者深入进行了这方面的研究,均认为受体是一些特殊结构,能与环境中相对应的物质(称配体)发生特异结合并产生一定生物效应。受体有膜上和膜内之分,它与配体的结合如同底物和酶,抗原和抗体的结合一样,具有高度特异性,高度亲和力,可逆性及饱和性。七十年代以来,由于同位素、荧光素标记及亚细胞成分的分离纯化等新技术的应用,使受体研究有了重大突破,不仅对受体可进行定位研究,还可进行定量分析,有的已深入到分子水平。随着受体研究的深入,新的受体不断被发现,其应用也不断扩大。如通过激素受体的检测,对阐明某些肿瘤的病因,预测肿瘤的预后,为治疗的选择提供了可靠依据。又如通过抗受体抗体的检测,对突眼性甲亢(Grave″s病),胰岛素抗性糖尿病,重症肌无力等自身免疫性疾病的发病机理有了新     

一氧化氮与微生物感染

本文简述一氧化氮在体内的合成和调节，阐述ＮＯ与病毒、细菌、菌和产产克次体感染，以及ＮＯ抗微生物感染的作用机制。     

PCT和CRP在感染性疾病诊断中的临床意义

目的:探讨PCT、CRP在感染性疾病诊断中的价值,评价两者与感染类型的相关性。方法:回顾性统计分析420例发热待查的患者血清PCT,血浆CRP浓度与感染的关系,采用胶体金法检测血清中PCT的浓度,免疫荧光比色法检测血浆中CRP浓度,血培养及血清学的方法检测感染是否存在及感染类型。结果:通过血培养或血清学鉴定出132例存在细菌感染(细菌组),病毒感染103例(病毒组),细菌病毒混合感染76例(混合感染组),109例阴性(非感染组)。PCT在细菌组水平最高(20.6±6.7)μg/L,非感染组水平最低(8.7±2.3)μg/L,四组间差异有统计学意义(F=34.69,P0.05)。CRP在混合组中水平最高(18.1±3.7)mg/L,在非感染组水平最低(5.8±1.7)mg/L,四组间差异有统计学意义(F=23.55,P0.05)。PCT对细菌感染检测的敏感度为92.7%,特异度为63.2%。PCT对病毒感染检测的敏感度为78.7%,特异性为46.0%。细菌感染组、病毒感染组和混合感染组血清CRP和PCT均呈正相关(P=0.000.05)。结论:细菌感染的PCT、CRP血清浓度明显高于病毒感染,两者对细菌感染患者的特异性均明显高于病毒感染,细菌感染PCT血清浓度较CRP变化敏感。     

流感病毒在Vero细胞系与MDCK细胞系增殖条件的比较

目的研究流感病毒H1N1及其他亚型在Vero细胞系和MDCK细胞系高效增殖的最适条件,比较两种细胞系对流感病毒的敏感性差异及影响敏感性差异的条件。方法在培养好的Vero细胞系与MDCK细胞系用不同的病毒感染复数(M.O.I)、胰酶浓度、病毒吸附时间、病毒维持液血清质量浓度等条件进行流感病毒在细胞上的增殖。结果在M.O.I为0.01接种流感病毒,吸附时间为1 h,胰酶质量浓度2μg/mL,血清质量浓度为8%时,流感病毒血凝素在MDCK细胞系可获得较高的滴度。结论 MDCK细胞系是适于流感病毒培养的细胞,它作为生产新型流感病毒疫苗的主要细胞基质需要进一步的研究。     

An Improved Strategy for Efficient Expression and Purification of Soluble HIV-1 Tat Protein in E.coli

Although the endogenous function of Tat has been elucidated in the past twenty years, the study of its exogenous activity has been hampered due to the difficulty of large scale preparation of the active Tat protein. To express the full-length Tat protein in E.coli, the tat gene was cloned from an HIV infected patient by overlapping PCR. Rare codon usage analysis showed that rare E.coli codons, especially consecutive rare codons for Arg, account for 14% （14 of 101） rare E.coli codons in the tat gene. The expression of the HIV-1 tat gene was verified to be very poor in strain BL21 （DE3） due to the abundance of rare codons; however, tat gene expression was found to be very efficient in the host strain of Rosetta-gami B （DE3）, which was supplemented with six rare tRNAs for Arg, Leu, Ile and Pro. Subsequent purification revealed that the proteins are soluble and unusually, the tagged Tat can form dimers independent of cystine disulfide bonds. The purity, integrity and molecular weight of the Tat protein were demonstrated by MALDI-TOF mass spectrometry. Reporter gene activating assay was further confirmed by investigating the transactivation activity of the recombinant Tat protein. Our improved strategy for efficient high level expression and purification of soluble Tat protein has paved the way to fully investigate its exogenous function.     

Midkine, a cytokine that inhibits HIV infection by binding to the cell surface expressed nucleolin

The growth factor midkine （MK） is a cytokine that inhibits HIV infection in cell cultures in an autocrine and paracrine manner by blocking the attachment of HIV particles to permissive cells. MK mRNA is systematically expressed in adult peripheral blood lymphocytes from healthy donors, while its expression becomes markedly but transiently increased upon in vitro treatment of lymphocytes with IL-2 or IFN-7 and activation of T lymphocytes by PHA or through the engagement of the CD28 antigen. The binding of MK to cells occurs specifically at a high and a low affinity binding site. This low affinity-binding site is the cell-surface expressed nucleolin, which is implicated in the mechanism of the initial attachment of HIV particles to cells. Accordingly, the nucleolin-binding HB-19 pseudopeptide has no effect on the MK binding to the high affinity binding site, whereas it prevents the binding of MK to the low affinity binding site, thus suggesting the low affinity receptor of MK is the cell-surface-expressed nucleolin. Confocal immunofluorescence laser microscopy revealed the colocalization of MK and the cell-surface-expressed nucleolin at distinct spots. The use of various deletion constructs of nucleolin then indicates that the extreme C-terminal end of nucleolin, containing repeats of the amino acid motif RGG, as the domain that binds MK. The specific binding of MK to the surface nucleolin is independent of heparan sulfate and chondroitin sulfate proteoglycans. After binding to cells, MK enters cells by an active process in which nucleolin and lipid rafts appear to be implicated. The potent and the distinct anti-HIV action of MK along with its enhanced expression in lymphocytes by various physiological stimuli, point out that MK is a cytokine that could be involved in HIV pathogenesis.     

丙型肝炎病毒感染相关的PRRs应答的分子机制

全球丙型肝炎病毒(HCV)感染者约为一亿七千万人,我国感染阳性率约为3·2%,HCV感染可导致肝炎、肝硬化、肝癌及肝外淋巴增生性疾病(包括巨球血症及B细胞淋巴瘤等)。成年HCV感染者超过70%的人发展为慢性持续性感染[1-3]。目前常用的抗病毒治疗的有效率不足50%,且费用昂贵[4]。由于HCV免疫应答及逃逸机制尚不十分清楚[5],使HCV疫苗的研制也举步维艰,已有的HCV感染给社会造成了严重负担。据预测未来几十年中国因HCV感染而死亡的人数仍将继续增长,因此必须加速研究HCV感染过程及宿主应答的分子机制,发现新的抗病毒治疗的药物靶标,研制…