抗脊髓灰质炎病毒Ⅱ型Sabin株D抗原单克隆抗体的研制

目的应用杂交瘤技术制备抗脊髓灰质炎病毒Ⅱ型Sabin株D抗原的单克隆抗体。方法用脊髓灰质炎病毒Ⅱ型Sabin株D抗原免疫雌性Balbc小鼠,取其致敏脾脏淋巴细胞与SP20小鼠骨髓瘤细胞融合。间接ELISA法筛选阳性克隆,制备腹水。检测McAb效价、抗体亚类和杂交瘤细胞核型。结果获得4株阳性杂瘤细胞株,选择一株制备腹水,ELISA效价为1∶2.4×106,中和试验效价1∶1.3×104,蛋白浓度为27.2mgmL。McAb与Ⅱ型Sabin株C抗原(ⅡC)、Ⅰ型Sabin株及Ⅲ型Phizer株D抗原(ⅠD、ⅢD)不发生交叉反应,具有高度特异性。结论成功的制备了针对脊髓灰质炎病毒Ⅱ型Sabin株D抗原的McAb,为进一步研究IPV效力检测奠定了基础。     

疱疹病毒感染和抗疱疹病毒药物研究进展

人类疱疹病毒感染为多发病和常见病,人群感染血清疱疹病毒抗体阳性率高。现有8种人类疱疹类病毒病毒：单纯疱疹1,2型,水痘-带状疱疹,E-B病毒,巨细胞病毒。疱疹病毒6,7和8型。可侵犯人体多种组织。引起皮肤,粘膜,淋巴,生殖和神经系统或肝,肺等赃器感染。其特点为潜伏性强,常形成隐性或急慢性局部或全身性感染。     

接种乙型肝炎疫苗后的HBs抗体反应和HLA的关系

接种乙型肝炎疫苗,不一定都产生HBs抗体,而且有人由于反应极低,在免疫过程中,却成了病毒携带者.本文为了研究这种HBs抗体产生不全状态与HLA的关系,分别检查了接种乙型肝炎疫苗小儿的HLA-A、B、C及DR基因位点.作者以观察一年以上的HBs抗原及HSe抗原阳性孕妇所生、采取了预防垂直感染措施的97例小儿为观察对象.小儿生后48小时内,静注乙肝免疫球蛋白(HBIG)及肌注HBIG,或单独肌注HBIG进行初次被动免疫,以达到预防分娩时HBV感染的目的.对达到预防感染效果的小儿,在生后第     

双烯雌酚人工抗原的合成及特异性抗体的制备

目的制备抗双烯雌酚(DIEN)特异性抗体,为进一步研制DIEN免疫检测试剂盒打基础。方法采用4-溴丁酸乙酯对DIEN进行活化,以活泼酯法与BSA偶联制备免疫原(DIEN-CP-BSA);经紫外扫描和飞行时间质谱扫描鉴定偶联情况;以DIEN-CP-BSA免疫Balb/c小鼠制备特异性抗体,间接(竞争)ELISA评价抗血清效价及特异性。结果本试验获得较高效价的DIEN抗血清,其效价达1∶64 000,双烯雌酚半抑制浓度(IC50)为62 ng/ml;抗血清与结构类似物己烷雌酚的交叉反应率仅为0.34%,与己烯雌酚和17-β雌二醇无交叉反应;利用该抗体建立的间接竞争ELISA检测法,双烯雌酚在10～300 ng/ml呈线性关系。结论本研究制备了抗双烯雌酚(DIEN)特异性抗体,为研究畜产品中DIEN残留及开发DIEN免疫检测试剂盒奠定了基础。     

H5亚型禽流感灭活疫苗病毒基因的定量检测及其与HI抗体效价之间关系的分析

目的建立H5亚型禽流感病毒灭活疫苗中病毒核酸的特异、敏感、快速的定量检测方法，分析其与HI抗体效价之间的相关性。探索禽流感灭活疫苗效力体外检验的方法。方法选取H5亚型禽流感病毒（avian influenza virus，AIV）血凝素（hemagglutinin，HA）基因保守序列，使用Prinler Express 2．0软件设计了特异性引物和TaqMan MGB（minor groove binder）探针，利用实时荧光PCR技术来定量检测禽流感灭活疫苗，通过体外转录，制备含有选定检测序列的RNA标准品，绘制标准曲线。对50批H5亚型禽流感灭活疫苗进行了荧光定量PCR检测，测定了疫苗的HA基因拷贝数，同时测定了50批疫苗相应的HI抗体效价。进行了疫苗的HA基因拷贝数与HI抗体效价之间相关性的分析。结果该方法的灵敏度为10拷贝，反应，标准曲线的相关系数为0．998003，对H9亚型禽流感灭活疫苗和其他禽病疫苗无交叉反应，特异性好、重复性佳。疫苗的HA基因拷贝数与HI抗体效价不相关。结论荧光定量PCR方法为H5亚型禽流感病毒灭活疫苗检测提供了一种特异、敏感、快速的定量检测方法。     

一对高亲和力抗人肌钙蛋白Ⅰ单克隆抗体的制备及鉴定

目的 获取抗人肌钙蛋白Ⅰ（cTnI）的单克隆抗体（McAb）.方法 以人cTnI作为抗原,免疫Balb/c小鼠,通过杂交瘤技术制备了抗人cTnI高亲和力、高特异性单克隆抗体.随后采用间接ELISA法测定抗血清效价,用protein G亲和纯化法纯化抗体,抗原竞争ELISA法鉴定抗体亲和力,SDS-PAGE法鉴定纯度,Western blotting鉴定抗cTnI单克隆抗体的特异性,竞争ELISA法分析抗原结合位点.结果 筛选出9株稳定分泌抗cTnI的单抗杂交瘤细胞株,其中A3、A9两株免疫球蛋白亚类均为IgG2a,分泌的抗体纯度高,与CK-MB、cTnT无交叉反应,效价均为：1：1024000,亲和力分别为4.21×10^8mol/L、1.07×10^8mol/L,抗原结合位点不同.结论 成功制备出了一对高亲和力、高特异性抗人cTnI单克隆抗体.     

重组EV71和CVA16型手足口病双价VLP疫苗构建及免疫保护效果评价

目的运用Bac-to-Bac杆状病毒表达系统表达EV71和CVA16的病毒样颗粒,并对纯化的重组EV71、CVA16双价病毒样颗粒疫苗进行免疫效果评价。方法构建重组杆状病毒Bacmid-P1-3CD,转染Sf9昆虫细胞,纯化EV71、CVA16的病毒样颗粒并检测其形态和生物特性;通过EV71、CVA16的病毒样颗粒免疫ICR雌鼠后,以EV71、CVA16强毒株腹腔攻击5日龄乳鼠,对重组EV71、CVA16型双价VLP免疫保护性进行评价。结果利用Bac-to-Bac杆状病毒表达系统成功构建并表达EV71和CVA16病毒样颗粒,颗粒大小约为23~30 nm,存在Mr约39 000 VP1特异性蛋白表达。重组EV71、CVA16双价VLP疫苗免疫接种能够诱导小鼠机体产生高效价的特异性抗体(EV71中和抗体效价为1∶960,CVA16中和抗体效价为1∶624)并发挥优于单价VLP疫苗的有效免疫保护效果。结论重组EV71、CVA16型双价VLP疫苗免疫原性和保护性显著高于单价疫苗,为手足口病疫苗的研发提供了新的思路及实验基础。     

应用蛋白微阵列同时检测人血清抗ToRCH抗体

目的：建立一种基于多元蛋白微阵列的免疫检测方法，用于同时检测人血清中识别弓形虫（TOXO）、风疹病毒（RV）、巨细胞病毒（CMV）、单纯疱疹Ⅰ型、Ⅱ型病毒（HSV-1、HSV-2）的特异性抗体。方法：将TOXO、RV、CMV、HSV-1和HSV-2抗原喷印到活化的玻片表面，抗原与活化基团共价结合后，制成蛋白微阵列。固定在玻片表面的抗原与病人血清中的特异性抗体反应、结合后，加入荧光标记的二抗，然后用高分辨率的激光共聚焦芯片扫描系统对蛋白微阵列进行扫描成像。所获得的图像用自行开发的软件进行分析，并自动判定并生成待测样本的定性结果。结果：使用此套蛋白微阵列系统检测抗TORCH抗体参考品，并与其它多种检测方法进行比较，各检测项目的符合率分别为92％～100％、92％～100％、96％、84％～96％、76％-96％。结论：多元蛋白微阵列法特异性强，灵敏度、准确度、精密度较高，具有应用和推广价值。     

柯萨奇病毒B 组5 型VP1 蛋白的外源表达及多克隆抗体的制备

目的:原核表达柯萨奇病毒B组5型的VP1蛋白,并制备其多克隆抗体及检测。方法:提取在Vero细胞中柯萨奇病毒B组5型的RNA,通过逆转录PCR(RT-PCR)扩增获取VP1基因序列,构建原核表达载体,大量诱导表达重组VP1蛋白。用His Trap HP亲和层析柱纯化重组蛋白,以纯化的目的蛋白为抗原,免疫雄性SD大鼠获得VP1蛋白多克隆抗体血清,ELISA测定该抗体的效价,微量中和实验测定抗体的中和活性,Western blot检测抗体的特异性,免疫组化检测抗体对组织中抗原的识别。结果:成功构建了VP1-pET-28a重组载体,并且在BL21细胞中成功诱导表达,亲和层析柱纯化后蛋白纯度大于90%。ELISA测得抗体的效价为1∶128 000,微量中和实验显示抗体没有中和活性,Western blot分析显示抗体能与CVA16和EV71交叉反应,免疫组化实验表明抗体能够识别组织中的CVB5抗原。结论:本研究成功制备了CVB5 VP1蛋白的高效价的多克隆抗体,为CVB5病毒疫苗和病毒诊断的研发提供了有效的工具。     

拟步甲科昆虫不同抗冻蛋白的免疫交叉性分析

目的 利用甲虫小胸鳖甲的抗冻蛋白MpAFP698制备抗血清,分析拟步甲科不同种类昆虫抗冻蛋白的免疫同源性.方法 通过DNA疫苗初次免疫-蛋白质加强的策略免疫新西兰大白兔,先以重组质粒pcDNA3-Mpafp698作为DNA疫苗免疫接种2次,再以融合蛋白His-MpAFP698进行蛋白质加强免疫2次.采用ELISA检测MpAFP698抗体的效价,Western blot法检测抗体的特异性及不同昆虫抗冻蛋白与MpAFP698抗体的免疫交叉反应.结果 兔抗血清的效价可达1∶40万,Western blot结果显示抗血清分别与His-MpAFP698及GST-MpAFP698蛋白有特异性结合,与小胸鳖甲抗冻蛋白MpAFP149、MpAFPS77以及来自不同昆虫的抗冻蛋白ApAPAFP914、OcAFP4有特异性结合.结论 荒漠地区不同甲虫的抗冻蛋白具有相同的抗原表位,可以发生免疫交叉反应.     

Obesity as disease and as social problem

This text is part of a broader line of study that aims to analyze how and why certain eating habits and bodily practices have become social problems, as is the case with fatness. We will show that the ideas that support the definition of obesity as a chronic and avoidable disease are leading experts and health authorities, and other social workers, to know and to think about its evolution in terms of "global" illness (epidemic) and to consider cultural factors as their main cause (obesogenic environment). Paradoxically, the international and national preventive measures taken are focused on changing individual behavior and, in particular, eating habits. The concepts about the regulation of excess weight and food provide interesting information about a particular understanding of lifestyles and culture and they take into account the current promotion of health patterns.     

Pharmacogenetic testing: not as simple as it seems

Pharmacogenetics has the potential to help guide treatment decisions by tailoring appropriate drugs and dosages to patients most likely to benefit. This straightforward clinical goal has led some to suggest that pharmacogenetic testing is free of ethical concerns. However, a number of potential risks and clinical uncertainties arise in considering the use of these new tools in clinical care. We propose a classification of pharmacogenetic tests to identify and prioritize the policy issues that will need to be addressed to ensure appropriate delivery of pharmacogenetic testing. We use the classification framework to consider the benefits and risks associated with ancillary information, timing of testing, and storage and retrieval of pharmacogenetic test results among health professionals. These issues have implications for informed consent and genetic counseling requirements, and for the role of health professionals.     

Human vascular tissues produce thromboxane as well as prostacyclin

Four different types of human blood vessels were examined for spontaneous and arachidonate-stimulated release of prostacyclin (PGI2) and thromboxane A2 (TXA2). Spontaneous PGI2 release was umbilical veins greater than umbilical arteries greater than saphenous veins greater than internal mammary arteries. With stimulation, PGI2 release by all four different vessels increased significantly (P less than 0.001) and was similar. Spontaneous release of TXA2 was also identified from all vessels examined and was similar. With stimulation, TXA2 release from all vessels increased significantly (P less than 0.001). Internal mammary artery released more TXA2 than other vessels on stimulation. The identity of TXA2 was confirmed by similar displacement of TXB2 antibody by TXB2 standards and by multiple dilutions of supernates. Treatment of vessels with aspirin inhibited PGI2 as well as TXA2 release, whereas treatment with OKY 1581 (TXA2 synthetase inhibitor) inhibited TXA2 release only. Two-dimensional thin-layer chromatography showed [14C]arachidonate conversion to PGI2 and TXA2 similar to that measured by radioimmunoassay. These data suggest that TXA2 is synthesized by human vessels. In older vessels when PGI2 generation is decreased, TXA2 production may by of pathophysiological significance.     

Guitarfish possess ipsilateral as well as contralateral retinofugal projections

Autoradiographic analysis of the primary retinal projections in the thornback guitarfish reveals both contralateral and ipsilateral projections to diencephalic, pretectal, and tegmental nuclei and the optic tectum. A total of 12 retino-recipient cell groups receive ipsilateral as well as contralateral inputs.     

Lyn deficiency affects B-cell maturation as well as survival

Lyn, an Src-family protein tyrosine kinase expressed in B lymphocytes, contributes to initiation of BCR signaling and is also responsible for feedback inhibition of BCR signaling. Lyn-deficient mice have a decreased number of follicular B cells and also spontaneously develop a lupus-like autoimmunity. We used flow cytometric analysis, BrdU labeling and our mathematical models of B-cell population dynamics, to analyze how Lyn deficiency impacts B-cell maturation and survival. We found that Lyn-deficient transitional 1 (T1) cells develop normally, but T2 cells develop primarily from the T1 subset in the spleen and fail to also develop directly from BM immature B cells. Lyn-deficient T2 cells either mature to the follicular B-cell type at a close to normal rate, or die in this compartment rather than access the T3 anergic subset. The ≈ 40% of WT follicular cells that were short-lived exited primarily by joining the T3 anergic subset, whereas the ≈ 15% Lyn(-/-) follicular cells that were not long lived had a high death rate and died in this compartment rather than entering the T3 subset. We hypothesize that exaggerated BCR signaling resulting from weak interactions with self-antigens is largely responsible for these alterations in Lyn-deficient B cells.     

Integral as well as proportional adrenal responses to ACTH

To characterize adrenal responses to ACTH, conscious dogs were infused for 40 min with normal saline or 20-1,500 ng/min alpha-ACTH. Plasma ACTH and corticosteroid levels were measured during the infusion and for 80 min after the end of the infusion. Half-maximal plateau corticosteroid responses were achieved with ACTH levels of 105 pg/ml (23 pM); a maximal plateau corticosteroid response (10.2 micrograms/100 ml) resulted from ACTH levels greater than 335 pg/ml (74 pM). When levels of ACTH between 335 and 3,000 pg/ml were achieved during the 40-min infusion, the duration of maximal corticosteroid levels was prolonged beyond the duration of the ACTH infusion. Plasma ACTH concentration decreased after the end of the infusion with a half-disappearance time of 3.6 +/- 0.2 min, but plasma corticosteroid concentrations and cortisol and corticosteroid secretory rates remained at or above the plateau secretory rates during the infusion and for about 80 min after the end of an infusion of 1,000 ng ACTH/min. When the total corticosteroid responses to the ACTH infusions were estimated, the relationship between the total corticosteroid response and the logarithm of the ACTH concentration was linear for 80-3,000 pg ACTH/ml.     

Peer review as professional responsibility: a quality control system only as good as the participants

The peer-review process remains a central part of the value and validity of scientific and technical publishing and proposal assessment. The peer review mechanism has many delicate components that should function most professionally and effectively for best results. An important central tenet is that all who seek to publish should freely avail themselves to review a commensurate load, considering many elements of professional conduct, ethics and responsibility in this process. The review itself should provide timely, unbiased, quality feedback to improve contributions to the system reviewers are serving. An additional component involves follow-on policing of published literature to assert its validity through consensus and validation. This short essay examines our collective duties as contributors, reviewers, and readers to the integrity and safekeeping of this essential quality control process.