抗人重组Cu－Zn超氧化物歧化酶单克隆抗体的制备及其在纯化抗原中的应用

本研究利用重组质粒ｐＣＺＳ转化大肠杆菌，制备出入重组Ｃｕ－Ｚｎ超氧化物歧化酶（ｒｈＣｕＺｎ－ＳＯＤ），进一步建立了３株鼠抗ｒｈＣｕＺｎＳＯＤ的单克隆抗体（ＭｃＡｂ）杂交瘤细胞株。经鉴定，它们分泌的ＭｃＡｂ分别属于ＩｇＧ１和ＩｇＧ２ａ亚类，能特异性地识别人ＣｕＺｎＳＯＤ，与ｐＣｚＳ空载菌蛋白无交叉反应。将此ＭｃＡｂ制备了免疫亲和层析柱，从大肠杆菌的粗提液中纯化出ｒｈＣｕＺｎＳＯＤ，ＳＤＳ－ｐＡＧＥ显示单一条带，纯度＞９８％，回收率５０％，酶化活达１０９７１．３Ｕ／ｍｇ蛋白。ｒｈＣｕＺｎＳＯＤ－ＭｃＡｂ的制备为ＣｕＺｎＪｉＤ产品的纯化和抗辐射等应用研究提供了有效的工具。     

HIV—1多表位膜蛋白在E.coli中的表达及在血清检测中的应用

目的：获得具有较多保守抗原表位的ＨＩＶ－１膜重组抗原，提高ＨＩＶ筛选ＥＬＩＳＡ试剂的质量。方法：利用分子克隆与ＰＣＲ方法，获得了新的ＨＩＶ－１膜重组抗原的基因克隆，编码ｇｐ１２０Ｃ端亲水区３７个氨基酸残基（ｅｎｖ４８２～５１８）以及ｇｐ４１膜外部分的１２８个氨基酸残基（ｅｎｖ５４８～６７５）。将该基因克隆到ｐＧＥＭＥＸ－１载体上融合表达，表达蛋白经纯化后，以１μｇ／ｍｌ的浓度包被ＥＬＩＳＡ板，用来     

几种抗人肺癌细胞单克隆抗体及其片段放射免疫显像的实验研究

对几种特异性强、灵敏度高的抗人肺癌细胞单克隆抗体（ＭｃＡｂ）及其片段，选择理想的放射性核素标记，为临床放射免疫显像（ＲＩＩ）提供实验依据。采用本室提供的３种ＩｇＧＭｃＡｂ：抗人大细胞肺癌ＭｃＡｂ（２Ｅ３）、抗人肺鳞状细胞癌ＭｃＡｂ（Ｓｍ１）、抗人肺腺癌ＭｃＡｂ（Ａｍ７）及２Ｅ３Ｆ（ａｂ′）２，中国科学院上海细胞所提供的ＩｇＭ抗人肺腺癌ＭｃＡｂ（ＬＣ１）及其片段，制备载人大细胞肺癌细胞株（ＰＬＡ８０１）或人肺腺癌细胞株（沪Ａ１）肿瘤裸鼠模型。注入１２５Ｉ、１３１Ｉ或９９ｍＴｃ标记的上述ＭｃＡｂ，行载瘤裸鼠显像。动态显像时应用感兴趣区法计算各时相Ｔ／ＮＴ值，并在注射ＭｃＡｂ后２４小时杀死动物，测定抗体生物分布。结果１３１Ｉ标记Ａｍ７、Ｓｍ１、２Ｅ３及ＬＣ１在２４小时载瘤裸鼠显影均良好。动态显像，各时相Ｔ／ＮＴ值及２４小时生物分布表明：９９ｍＴｃ标记ＭｃＡｂ优于１３１Ｉ标记ＭｃＡｂ；抗体片段优于完整抗体；单抗在同型抗原肿瘤的浓聚高于异型抗原肿瘤。故临床肿瘤ＲＩＩ核素采用９９ｍＴｃ、抗体选择混合抗体为好。     

抗内毒素核心区单克隆抗体及其对革兰阴性菌脓毒性休克…

通过大肠杆菌Ｒｃ突变株（Ｊ５）免疫的小鼠脾细胞和Ｓｐ２／０骨髓瘤融合，建立了２２个抗内毒素单克隆抗体细胞株。对７株进行了Ｉｇ类型鉴定：５株为ＩｇＭ／ｋ，１株为ＩｇＧ２ｂ／λ，１株为ＩｇＧ３／λ。５株ＩｇＭ抗体识别的表位均在内毒素的内核心区经沉淀法和凝胶过滤法纯化腹水，抗体纯度达９０％。在Ｅ．ｃｏｌｉ临床分离物或Ｓ－ＬＰＳ所致的小鼠休克模型上，腹腔注射抗体６２．５－１２５μｇ／只，实验组存活率比对照     

利用噬菌体表面呈现技术制备HIV-1整合酶单链抗体的初步研究

目的：研制ＨＩＶ－１整合酶（ｉｎｔｅｇｒａｓｅ，ＩＮ）的重组噬菌体单链抗体。方法：采用噬菌体表面呈现方法。结果：由ＩＮ蛋白免疫的小鼠脾细胞ｍＲＮＡ中构建了单链抗体基因ｃＤＮＡ文库，克隆入噬菌粒载体ｐＣＡＮＴＡＢ５Ｅ，经转化大肠杆菌ＴＧ１，获得了库容为３．５×１０５的表达文库。利用包被在酶联板上的ＩＮ蛋白进行四轮亲和富集，筛选出７６株具有ＩＮ结合活性的重组噬菌体。对随机挑选的一株克隆进行了ＤＮＡ序列分析，证明其符合小鼠抗体序列。结论：获得了抗ＨＩＶ－１ＩＮ的噬菌体单链抗体，为其进一步研究打下基础。     

亲和素—生物素系统^153Sm标记物在抗CEA单抗预定位放免显 …

目的 探讨＾１２５Ｓｍ标记亲和素－生物素系统（ＡＢＳ）在荷人结肠癌裸鼠模型中进行抗ＣＥＡ单克隆抗体（ＭｃＡｂ）预定位放免显像的可行性。方法 首先用＾１５３Ｓｍ标记螯合ＤＴＰＡ生物素（ＤＢ２）,然后利用生物素与亲和素（ＡＶ）的高亲和力特性,对链霉亲和素（ＳＡ）进行＾１５３Ｓｍ标记,通过ＤＴＰＡ环酐法将＾１５３Ｓｍ标记到抗ＣＥＡ ＭｃＡｂ上,应用三步法方法Ａ（生物素化ＣＥＡ ＭｃＡｂ－ＡＶ－＾１５３Ｓ     

丙型肝炎病毒核心蛋白可溶性单链可变区抗体在大肠杆菌中的表达

利用分子生物学技术,构建表达丙型肝炎病毒（ＨＣＶ）核心蛋白的人源单链可变区抗体（ＳｃＦｖ）的原核表达载体,并在大肠杆菌ＪＭ１０９中表达可溶性的ＨＣＶ－ｃｏｒｅ－ＳｃＦｖ。以重组的ＨＣＶ核心蛋白为包被抗原,利用噬菌体抗体库的表面展示技术,筛选到含有ＨＣＶ－ｃｏｒｅ－ＳｃＦｖ基因的噬菌体克隆。从噬菌体抗体阳性克隆中提取质粒,经ＮｃｏⅠ／Ｎｏｔ Ⅰ酶切鉴定,该ＳｃＦｖ基因由７５０ｂｐ组成。将其亚克隆到ｐＣＡＮＴＡＢ５Ｅ表达载体中,转化大肠杆菌ＪＭ１０９,提取质粒进行ＤＮＡ序列测定,符合ＳｃＦｖ的重链可变区和轻链可变区基因结构特点。ＩＰＴＧ诱导转化的大肠杆菌ＪＭ１０９,在其培养上清中获得了可溶性ＨＣＶ－ｃｏｒｅ－ＳｃＦｖ的表达。酶联免疫吸附法（ＥＬＩＳＡ）证实表达的ＨＣＶ－ｃｏｒｅ－ＳｃＦｖ进行聚丙烯酰胺凝胶电泳（ＰＡ     

阳江高本底地区居民中免疫水平与对病毒免疫反应的研究

探讨在天然高本底电离辐射作用下机体的免疫水平与对病毒的免疫反应。方法利用单克隆抗体ＡＰＡＡＰ法检测白细胞介素－２分泌细胞（ＩＬ－２ＳＣ）；利用间接免疫酶法检测抗ＥＢ病毒抗原抗体水平（ＶＣＡ／ＩｇＡ，ＶＣＡ／ＩｇＧ，ＥＡ／ＩｇＡ，ＥＡ／ＩｇＧ）；采用酶标免疫吸附沉淀法检测乙型肝炎（ＨＢＶ）表面抗原（ＨＢｓＡｇ），表面抗原抗体（ＨＢｓＡｂ）和核心抗体（ＨＢｃＡｂ）。结果高本底地区（ＨＢＲＡ）居民外周血淋巴细胞ＩＬ－２ＳＣ频数显著高于对照（ＣＡ）；ＶＣＡ／ＩｇＡ、ＶＣＡ／ＩｇＧ、ＥＡ／ＩｇＧ抗体水平均高于ＣＡ，但仅ＶＣＡ／ＩｇＧ抗体滴度水平存在差异显著性，两地区ＶＣＡ／ＩｇＡ≥１５阳性检出率均显著高于广东地区的平均水平；ＨＢＶ感染率与ＨＢｓＡｂ阳性率为ＨＢＲＡ高于ＣＡ（Ｐ＞０．０５）。结论提示长期低水平电离辐射作用机体免疫功能有增强的趋势。     

抗内毒素核心区单克隆抗体及其对革兰阴性菌脓毒性休克症小鼠保护作用的研究

通过大肠杆菌Ｒｃ突变株（Ｊ５）免疫的小鼠脾细胞和Ｓｐ２／０骨髓瘤融合，建立了２２个抗内毒素单克隆抗体细胞株。对７株进行了Ｉｇ类型鉴定：５株为ＩｇＭ／Ｋ，１株为ＩｇＧ２ｂ／λ，１株为ＩｇＧ３／λ。５株ＩｇＭＪｉ体识别的表位均在内毒素的内核心区。经沉淀法和凝胶过滤法纯化腹水，抗体纯度达９０％。在Ｅ．ｃｏｌｉ临床分离物或Ｓ－ＬＰＳ所致的小鼠休克模型上，腹腔注射抗体６２．５～１２５μｇ／只，实验组存活率比对照组高３７．５％～７５．０％。体外结合和抑制试验表明，这些抗体与克雷伯杆菌、沙雷茵、鼠伤寒杆菌、痢疾杆菌、绿脓杆菌等属有很强的交叉反应，提示本组抗体可能对革兰阴性菌感染有广谱的保护作用。     

乙型肝炎病人血清中PreS1／2抗原检测方法的建立及初步应用

目的：建立乙型肝炎病毒（ｈｅｐａｔｉｔｉｓＢｖｉｒｕｓ，ＨＢＶ）ＰｒｅＳ抗原检测方法。方法：表达与纯化了ＨＢＶＰｒｅＳ１／２抗原，并用纯化蛋白免疫家兔，利用纯化抗血清包被ＥＬＩＳＡ板，吸附病人血清中包含ＰｒｅＳ１／２抗原的ＨＢＶ的病毒颗粒或病毒亚单位，特异吸附颗粒用抗ＨＢｓＡｇ的酶标单抗检测。结果：利用金属螯合柱层析的方法高度纯化了ＰｒｅＳ重组抗原，用复性的重组抗原免疫家兔３次，血清抗体滴度可达１     

人免疫缺陷病毒HB-J株的分离和生物学特性研究

目的：分离和鉴定我国流行的人免疫缺陷病毒（ｈｕｍａｎ ｉｍｍｕｎｏｄｅｆｉｃｉｅｎｃｙ ｖｉｒｕｓ,ＨＩＶ）毒株,研究我国ＨＩＶ毒株的生物学特性。方法：周外用血单个核细胞（ｐｅｒｉｐｈｅｒａｌ ｂｌｏｏｄ ｍｏｎｏｎｕｃｌｅａｒ ｃｅｌｌｓ,ＰＢＭＣ）共培养的方法分离ＨＩＶ,用观察细胞病变、检测Ｐ２４抗原和前病毒ＤＮＡ的方法监测病毒生长的情况。结果：在建立共培养的第１２天观察到明显的细胞病变,主要表面为大泡样细胞和多核巨细胞。接种第９天细胞培养上清液的Ｐ２４抗原由阴性转为阳性,并且浓度迅速增加,接种第１２天浓度即达到１７８０ｐｇ／ｍｌ,第１４天达到峰值,为２３２０ｐｇ／ｍｌ。但该株病毒的ＰＢＭＣ培养上清液转种于ＭＴ４细胞后,培养上清液Ｐ２４抗原和前病毒ＤＮＡ始终为阴性且不出现细胞病变。结论：用ＰＢＭＣ共培养的方     

第四代HIV诊断试剂检测HIV抗体假阳性原因分析

 目的分析第四代HIV诊断试剂检测HIV抗体假阳性的原因,减少假阳性检出率。方法选取119例HIV四代试剂检测初筛阳性的标本,送HIV确认实验室确认HIV抗体结果,分析比对两者的结果,探讨出现假阳性的原因。结果119例HIV抗体初筛阳性的标本,送至确认实验室回报结果40例HIV抗体阳性,28例HIV抗体不确定,51例HIV抗体阴性。随着S/CO值的增高,初筛结果与确证结果阳性符合率也随之升高。结论HIV四代检测试剂假阳性率较高,这与试剂同时检测HIV抗体和P24抗原有关。当1相似文献   

与HIV-1共感染的HHV-6毒株的分离、传代培养与鉴定

目的：对一株从中国河南省一名艾滋病患者体内分离的、可在MT4细胞上稳定传代的未知病毒29A进行鉴定，确定该病毒种类及型别。方法：用从该患者外周血单核细胞中分离的病毒株感染MT4细胞，获得可在该细胞上稳定传代的病毒株29A。通过细胞病变特征、电镜观察结果、HIV-1 p24抗原检测以及HIV-1 pol区基因扩增结果，对该毒株是否为HIV-1进行鉴别。在电镜下对病毒感染细胞的超薄切片进行观察，发现该病毒呈现疱疹病毒形态特征，因此设计人类疱疹病毒6型（HHV-6）U69、U16／U17、U60／U66三个不同基因片段的特异性引物并进行巢式PCR扩增，对扩增产物进行序列测定和结果分析。结果：该毒株HIV-1 p24抗原检测为阴性；用HIV-1 pol区特异性引物未扩增出目的片段；细胞病变形态不同于HIV-1引起的病变；电镜观察该病毒直径为160～200nm，明显大于HIV—1，表明该传代株不是HIV-1。用人类疱疹病毒6型（HHV-6）U69、U16／U17、U60／U66等基因的特异性引物均扩增出了目的片段，序列分析的结果表明该毒株为HHV-6病毒B亚型。结论：在国内首次从艾滋病患者的外周血淋巴细胞中分离出HHV-6，为进一步研究HHV-6和HIV-1的相互作用奠定了基础。     

Benefits of exercise training in Spanish prison inmates

Prison populations are growing in Western countries. Imprisoned people usually have a poor health status and an increased risk to suffer chronic debilitating conditions as coinfection with the HIV and hepatitis C virus (HCV) and/or opioid dependency. We studied the effects of a 4-month concurrent cardiorespiratory and resistance training program on the cardiorespiratory fitness, lower and upper body dynamic strength endurance (6-RM test for bench press and knee-extensor exercise, respectively), muscle mass and quality of life (QOL) of adult prison inmates who are HIV/HVC co-infected and enrolled in a methadone maintenance program (n = 9; mean [SD] age: 37 [3] yrs). We also evaluated a control group (n = 10; 37 [2] yrs). A significant combined effect of group and time was found for peak completed workload (W) (p < 0.01), peak heart rate (HR (peak)) (p < 0.05) and rate of HR decrease at 1-min postexercise compared to HR (peak) (p < 0.05), respectively, in a gradual cycle ergometer test. A significant combined effect of group and time was also found for both bench press and knee-extensor 6-RM tests, respectively (p < 0.05). Supervised exercise training can improve the overall physical fitness of incarcerated people. Our results suggest that this type of intervention could be applied in prisons of Western societies.     

AIDS awareness in the conduct of radiologic procedures: guidelines to safe practice

The epidemic of the acquired immunodeficiency syndrome (AIDS) has affected all geographic regions of the United States. Indeed, it is likely that a majority of U.S. hospitals have cared for patients who are infected with the human immunodeficiency virus (HIV), the virus that causes AIDS. More than 46,000 individuals with HIV infection have fulfilled the strict diagnostic criteria for AIDS (1). All of these patients with AIDS have been seriously ill and have been hospitalized, often multiple times and for prolonged periods. Another, larger group patients with HIV infection has had another form of the disease, the so-called AIDS-related complex. Many of these patients have also spent time in hospitals. An even larger group of individuals has been infected with HIV but has remained asymptomatic to date. Some of these persons have been admitted to hospitals or have received outpatient diagnostic procedures for intercurrent medical or dental complaints that were unrelated to their asymptomatic HIV infection. At the time of their medical evaluation, some of these asymptomatic individuals were known to have a positive HIV antibody test. It is likely, however, that the serologic status of the majority of asymptomatically infected patients was unknown. Thus, by now a very large number of health-care personnel in the United States, including diagnostic radiology staff, have assisted in the care of HIV-infected patients.     

Arthritis associated with HIV infection: radiographic manifestations

Radiographs of symptomatic joints were retrospectively evaluated in 24 patients with inflammatory arthritis and human immunodeficiency virus (HIV) infection. Clinically, 20 patients had a seronegative arthritis including Reiter syndrome (54%), psoriatic arthritis (17%), and undifferentiated forms of spondyloarthropathy (13%). These patients were indistinguishable radiographically from patients with typical seronegative disorders except for the predominance of lower-extremity abnormalities. Four patients (17%) had a rheumatoidlike arthritis defined as acute symmetric polyarthritis (ASP). With the exception of extensive proliferative periostitis, ASP simulated classic rheumatoid arthritis. HIV-associated arthritis was manifest during various stages of HIV infection. It preceded acquired immunodeficiency syndrome in 64% of patients with stage IV HIV infection. Awareness of the coexistence of HIV infection in patients with the above-mentioned arthritides is important, since immunosuppressive therapy, commonly used in the treatment of arthritis, can have detrimental effects in patients with HIV infection.     

Human immunodeficiency virus and the emergency setting: Legal considerations

The emergence of human immunodeficiency virus (HIV) has changed the atmosphere surrounding the provision of emergency medical treatment. HIV-positive patients fear denial of needed health services based on their seropositivity, whereas medical providers worry about occupational exposure. Both patients and providers have concerns regarding HIV testing and the confidentiality of test results. This article summarizes some of the many legal implications created by HIV in the emergency setting.     

大肠癌组织Fas抗原和p53蛋白的表达及临床意义

目的　探讨大肠癌组织Fas抗原及 p5 3蛋白的表达及临床意义。 方法　采用免疫组织化学方法对 32例大肠癌组织的Fas抗原 ,16例正常大肠组织及 p5 3蛋白表达进行检测和比较。 结果　Fas抗原在正常大肠组织中表达阳性 ,p5 3蛋白在正常大肠组织中表达阴性 ,Fas抗原在大肠癌组织中表达阳性率为 4 0 .6 2 %(13/ 32 ) ,p5 3蛋白在大肠癌组织中的表达阳性率为 5 3.13%。Fas抗原及 p5 3蛋白在大肠癌组织中的表达与患者性别、年龄、肿瘤大小、部位、大体分型、浸润深度、Ducks及局部淋巴结转移等无明显关系 (P >0 .0 5 ) ,Fas抗原在大肠癌组织明显下降 ,Fas抗原表达与大肠癌组织学类型及肝转移相关 (P <0 .0 5 )。p5 3蛋白表达与大肠癌肝转移相关 (P <0 .0 5 )。结论　Fas抗原在大肠癌组织中表达明显下降。随着肿瘤恶性程度的提高 ,Fas抗原表达减少。在肝转移患者中未见表达。p5 3蛋白在大肠癌组织中表达阳性 ,尤以发生肝转移者为著。Fas抗原表达下降或缺乏及 p5 3蛋白过度表达均提示预后不良     

CyberKnife stereotactic irradiation for metastatic brain tumors

BACKGROUND: The CyberKnife provides a new technique for performing frameless stereotactic irradiation. So far, few reports have been published on clinical outcomes obtained with the CyberKnife. This report summarizes our clinical experience with CyberKnife irradiation for metastatic brain tumors. MATERIALS AND METHODS: Seventy-seven lesions (48 patients) were evaluated and analyzed, and 66 lesions in 41 patients were treated with stereotactic radiosurgery (SRS). The prescribed dose was 9 to 30 Gy. RESULTS: Freedom from progression of the tumors was more likely with a prescribed dose of at least 24 Gy than with one of less than 20 Gy (p=0.0244; log-rank test). The CR (complete response) rate was significantly higher when D99 was at least 24 Gy (p=0.0045). There were no severe side effects. CONCLUSION: Stereotactic irradiation with the CyberKnife for metastatic brain tumors is effective and safe. D99 should be at least 24 Gy for CyberKnife SRS treatment.     

河南省驻马店地区HIV感染率的调查分析

目的　通过对来我院就医的抗 HIV阳性患者的定群分析 ,初步了解驻马店地区HIV的感染率和传播方式。方法　应用抗 HIVELISA试剂盒进行抗 HIV检测。结果　 3年半来共检测标本 12 980例其中检出阳性标本 15 0例 ,阳性率为 1 15 % ,经过调查发现 84%的阳性患者 1995年以前有过卖血或血浆史。结论　我区HIV感染者正处在发病阶段 ,并且感染疫情有爆发的潜在危险。