人狂犬病毒IgG抗体定量测定试剂盒(酶联免疫法)的评价

目前,被国际兽疫局（OIE）和世界卫生组织（WHO）狂犬病专家委员会认可的检测狂犬病病毒血清／血浆抗体的方法有小鼠中和试验（MNT）、快速荧光灶抑制试验（RFFIT）、荧光抗体病毒中和（FAVN）试验口,但均存在耗时长、操作繁琐、需要昂贵荧光显微镜以及专业人员培训等缺点,只能在有条件的实验室进行。而抗狂犬病毒抗体的ELISA测定方法因其特异、     

狂犬病毒核蛋白抗原ELISA定量法的建立及初步验证

目的建立检测狂犬病疫苗各工序产品中核蛋白抗原含量的双抗体夹心ELISA法。方法以兔抗狂犬病毒多抗为包被抗体,辣根过氧化物酶标记的抗狂犬病毒核蛋白单抗作为酶标记抗体,对狂犬病毒核蛋白抗原含量进行定量测定,并对该方法进行初步验证。结果此方法线性相关系数大于0.97;最佳线性范围为0.000625～0.01IU/ml,定量限度为0.000625IU/ml;准确性为102%～109%;变异系数(CV)为7.2%～9.4%;与小牛血清、牛血清清蛋白(BSA)、卵清蛋白(OVA)、流感疫苗纯化液、乙脑疫苗纯化液、甲肝疫苗纯化液等均无交叉反应。结论该方法特异性强,灵敏度高,准确性、重复性和稳定性好,可用于狂犬病疫苗各工序产品中核蛋白抗原的定量检测。     

定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定方法的建立

目的 建立定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定(ELISA)方法.方法 选择包被抗原和酶标二抗的最适浓度,用人狂犬病毒抗体标准品标化系列工作标准品,对临床收集的各类血清标本进行检测和分析.结果 间接ELISA定量检测中和抗体的最佳包被糖蛋白抗原浓度为10μg/L,酶标二抗的工作浓度为1:4 000,临床检验,间接ELISA定量方法灵敏度为100Vo.特异度为98.2%,总符合率99.0%;与快速荧光灶抑制实验呈(RFFIT)高度相关性(r=0.981).结论 本研究建立的定量检测狂犬病中和抗体方法,可以用于人狂犬病毒中和抗体的定量检测.     

639例狂犬病疫苗免疫后抗体检测结果与分析

为了解狂犬病疫苗对人体免疫水平的影响，本文对1996年登记在册的有动物（犬与猫）咬伤史者639例进行抗狂犬病毒抗体检测，现将结果报告如下。116床资料本组639例中，男432例，女207例，年龄3～60岁。按狂犬病疫苗常规免疫程序，干咬伤后的当日至3日内，在1月内完成5针上臂三角肌肌注，并于免疫后第25～30天，采静脉血Zml，分离血清，进行抗狂犬病病毒抗体检测。凡抗体检测为阳性的血清，按1：5、l：10的滴度，继续做抗体效价检测，以观察人体免疫后的保护效价，及不同性别、不同年龄组人群免疫效价的情况。狂犬病疫苗由上海生物制品研究所…     

柯萨奇病毒A16型快速纯化和中和性单克隆抗体制备与鉴定

目的建立柯萨奇病毒A16型快速纯化方法,制备中和性单抗,并对单抗进行分析。方法收获CA16培养上清液,超滤浓缩,氯化铯密度梯度离心纯化病毒颗粒,透射电镜鉴定纯化产物。福尔马林灭活CA16,免疫BALB/c小鼠,制备分泌抗CA16特异性单抗的杂交瘤细胞系,用ELISA和中和试验分别对单抗特性进行分析。结果初步建立CA16病毒氯化铯密度梯度纯化方法,电镜显示,病毒颗粒为二十面体立体对称球形结构,病毒直径在20~30nm间,大小均匀。获得2株分泌抗CA16单抗的杂交瘤细胞系,2株单抗均为IgG2a亚型,Anti/CA16/5效价为103,Anti/CA16/10效价为104。2株抗体的中和效价分别为1∶256和1∶1 024。结论初步建立氯化铯密度梯度纯化CA16的方法,筛选出2株具有中和活性的抗CA16单抗,为CA16病毒的基础研究提供重要的原材料。     

抗-CD47单克隆抗体干扰输血相容性检测1例

目的 探讨抗-CD47单抗对输血相容性检测的影响及处理措施。方法 对1名具有抗-CD47单抗用药史的患者血液标本进行输血相容性检测，分析抗-CD47单抗效价，鉴定巯基试剂对抗-CD47单抗的洗脱效果，利用中和抗体及抗球蛋白筛选与患者相配合的血液成分。结果 患者血型鉴定为A型，RhD(+),不规则抗体筛查、DAT试验、交叉配血均为3+或4+。利用中和抗体或Gamma-clone抗球蛋白均可为患者筛选到主侧配血相合的红细胞。患者输注后无不良反应，贫血状况得到明显改善。结论 利用中和抗体或Gamma-clone抗球蛋白处理患者血液标本，可以消除抗-CD47单抗对输血相容性检测的影响，保障临床用血的安全性、及时性和有效性。     

基于Vero E6细胞的水痘-带状疱疹病毒抗膜抗原荧光抗体试验的建立与评价

目的 建立并评价基于非洲绿猴肾细胞(Vero E6)的水痘-带状疱疹病毒(VZV)抗膜抗原荧光抗体(FAMA)试验方法。方法 在VZV-Oka减毒活疫苗株在Vero E6细胞中的适应培养毒株VZV-Oka-E6的基础上,首先使用3个不同感染剂量(10^4.65、10^4.95和10^5.25 TCID₅₀)的VZV-Oka-E6病毒株感染Vero E6细胞,于显微镜下观察病变情况确定最佳的VZV-Oka-E6病毒株感染量;然后对比使用不同固定液固定感染细胞后制备的固定抗原片可检测到的灵敏度确定最优的固定液,从而建立基于Vero E6细胞的中和抗-VZV检测FAMA试验方法;采用建立的FAMA法,检测不同效价的VZV免疫球蛋白国际标准品,确定该方法的敏感性;检测人单纯疱疹病毒(HSV)1、2型特异性抗体,评价该方法的特异性;分别使用同一批次制备和4个不同批次制备的VZV感染细胞固定抗原片检测VZV免疫球蛋白国际标准品的中和抗-VZV,确定该方法的批内和批间重复性;检测已知的3种效价的VZV免疫球蛋白国际标准...     

用Rh(D)阳性红细胞吸收制备抗-D试剂血清

目的　解决人源Rh试剂血清制备困难 ,节约Rh阴性红细胞。方法　①将A、B型含有抗 D的 2种血浆混合 ,经过稀释中和后的血浆完全抗体效价下降到只有 2 ;②采用D抗原位点结合被完全封闭的相应ABO血型Rh(D)阳性红细胞 ,吸收中和后的抗 A、抗 B。结果　既吸收了抗 A、抗 B ,又未使抗 D效价降低 ,检测效价及特异性符合Rh抗 D试剂血清要求。结论　采用中和后再吸收的方法 ,解决了人源Rh试剂血清制备困难 ,可节约大量Rh阴性红细胞。     

利用脐血分析O型血清的抗-AB

目的：探讨O血清抗AB的抗体性质和效价。方法：分别用脐血红细胞和成人红细胞作为吸收细胞完成吸收放散试验。利用A（B）细胞吸收O血清，然后采用适当的放散方法对放散液的抗B（A）的效价进行测定。结果：利用脐血作为吸收细胞，4℃吸收放散试验抗AB效价为4－32，37℃吸收放散试验抗AB效价为4－16；利用成人细胞作吸收放散试验，4℃抗AB效价为4－16，37℃吸收放散试验抗AB效价为0－4。两种温度下利用脐血细胞与利用成人细胞作为吸收细胞吸收放散试验的结果的差异有显性（P＜0．01）。结论：O血清抗AB有IgM性质，也有IgG性质，IgM性质抗AB效价为4－32，IgG性质抗AB效价为4－16。     

改良抗体结合试验的建立和在灭活狂犬病疫苗效价检测中的应用

目的对抗体结合试验(ABT)进行改进并将其用于灭活狂犬病疫苗效力的测定。方法将快速荧光灶抑制试验(RFFIT)与ABT方案融合,在疫苗样品稀释、剩余中和抗体检测、结果计算方面进行改进,形成改良抗体结合试验(M-ABT);应用M-ABT对10种灭活狂犬病疫苗随库存时间增加疫苗效力改变的情况进行检测。结果 M-ABT可以在28 h内完成,比ABT节省2 d,检测结果与人用狂犬病疫苗效价测定法(NIH)基本等效;10种疫苗随着库存时间增加疫苗效力下降。结论 M-ABT法能够成功建立,并用于灭活狂犬病疫苗效力检测效果较满意。     

Migraine and genetic and acquired vasculopathies

It is remarkable that migraine is a prominent part of the phenotype of several genetic vasculopathies, including cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL), retinal vasculopathy with cerebral leukodystrophy (RVCL) and hereditary infantile hemiparessis, retinal arteriolar tortuosity and leukoencephalopahty (HIHRATL). The mechanisms by which these genetic vasculopathies give rise to migraine are still unclear. Common genetic susceptibility, increased susceptibility to cortical spreading depression (CSD) and vascular endothelial dysfunction are among the possible explanations. The relation between migraine and acquired vasculopathies such as ischaemic stroke and coronary heart disease has long been established, further supporting a role of the (cerebral) blood vessels in migraine. This review focuses on genetic and acquired vasculopathies associated with migraine. We speculate how genetic and acquired vascular mechanisms might be involved in migraine.     

Fibrinogen and fibrin structure and functions

Fibrinogen molecules are comprised of two sets of disulfide-bridged Aalpha-, Bbeta-, and gamma-chains. Each molecule contains two outer D domains connected to a central E domain by a coiled-coil segment. Fibrin is formed after thrombin cleavage of fibrinopeptide A (FPA) from fibrinogen Aalpha-chains, thus initiating fibrin polymerization. Double-stranded fibrils form through end-to-middle domain (D:E) associations, and concomitant lateral fibril associations and branching create a clot network. Fibrin assembly facilitates intermolecular antiparallel C-terminal alignment of gamma-chain pairs, which are then covalently 'cross-linked' by factor XIII ('plasma protransglutaminase') or XIIIa to form 'gamma-dimers'. In addition to its primary role of providing scaffolding for the intravascular thrombus and also accounting for important clot viscoelastic properties, fibrin(ogen) participates in other biologic functions involving unique binding sites, some of which become exposed as a consequence of fibrin formation. This review provides details about fibrinogen and fibrin structure, and correlates this information with biological functions that include: (i) suppression of plasma factor XIII-mediated cross-linking activity in blood by binding the factor XIII A2B2 complex. (ii) Non-substrate thrombin binding to fibrin, termed antithrombin I (AT-I), which down-regulates thrombin generation in clotting blood. (iii) Tissue-type plasminogen activator (tPA)-stimulated plasminogen activation by fibrin that results from formation of a ternary tPA-plasminogen-fibrin complex. Binding of inhibitors such as alpha2-antiplasmin, plasminogen activator inhibitor-2, lipoprotein(a), or histidine-rich glycoprotein, impairs plasminogen activation. (iv) Enhanced interactions with the extracellular matrix by binding of fibronectin to fibrin(ogen). (v) Molecular and cellular interactions of fibrin beta15-42. This sequence binds to heparin and mediates platelet and endothelial cell spreading, fibroblast proliferation, and capillary tube formation. Interactions between beta15-42 and vascular endothelial (VE)-cadherin, an endothelial cell receptor, also promote capillary tube formation and angiogenesis. These activities are enhanced by binding of growth factors like fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF), and cytokines like interleukin (IL)-1. (vi) Fibrinogen binding to the platelet alpha(IIb)beta3 receptor, which is important for incorporating platelets into a developing thrombus. (vii) Leukocyte binding to fibrin(ogen) via integrin alpha(M)beta2 (Mac-1), which is a high affinity receptor on stimulated monocytes and neutrophils.     

Telemedicine and teleradiology technologies and applications

Summary. Telemedicine and teleradiology hold the key for improving future health care delivery. In this paper we first review current communication and computer technologies used in telemedicine and teleradiology. Five examples in teleradiology applications are given including hospital-integrated picture archiving and communication systems, tele-neuro-imaging, telemammography, university consortium teleradiology service, and teleradiology for second opinion. Parameters important to teleradiology applications like costs, image quality, system reliability, and turn around time are considered. Data security is discussed, including patient confidentiality and image authenticity-which will be a major issue in future teleradiology applications.     

创伤高血糖与感染和MODS早期诊断和干预

本文详细介绍了创伤后血糖应激适度理论,以及高血糖与感染和多器官功能不全综合征的关系;提出涉及胰岛B细胞功能不全的MODS实验诊断新方案和极化液个体化干预新措施,可早期发现创伤MODS、降低感染率及MODS发生率和病死率。     

腹膜后纤维化与肾积水发生关系的临床研究

目的：探讨腹膜后纤维化（RPF）导致肾积水的原因及诊治经验。方法：回顾分析2004年1月—2010年12月24例腹膜后纤维化致肾积水患者的诊治资料。结果：（1）RPF患者常见首发症状为腰背痛或腹痛（69.2%）;（2）红细胞沉降率（ESR）增快和血清IgG4升高最常见。超声检查仅提示上尿路积水。RPF的静脉肾盂造影（IVP）和CT尿路成像（CTU）表现具有特征性。IVP肾盂输尿管显影不良时,CTU能较清晰的显示上尿路影像。CT扫描发现腹膜后软组织肿块9例（37.5%）,优于超声检查;（3）输尿管松解和腹腔化手术治疗22例;行肾切除术1例;行输尿管置双J管术1例。最终确诊为继发性RPF8例,其中4例为术前诊断,3例为术中腹膜后软组织肿块冷冻活检证实,1例为术后病理证实;（4）特发性RPF手术后肾积水均获长期缓解,而继发性RPF的预后取决于原发疾病及其治疗方案。结论：影像学检查是诊断RPF的重要手段,CTU优于超声检查和IVP。输尿管松解和腹腔化手术可以使特发性RPF输尿管梗阻得到长期的缓解,术中对肿块进行冷冻活检有助于鉴别特发性和继发性RPF,及时调整治疗方案。     

探讨儿童慢性顽固性咳嗽与支原体感染的关系及临床治疗观察

目的探讨儿童慢性顽固性咳嗽与肺炎支原体（MP）感染的关系及临床疗效观察。方法采用回顾性研究方法对于现将2005年3月至2008年3月在我院的55例确诊慢性顽固性咳嗽患儿,主要表现为肺炎支原体感染为临床特点进行分析,并进一步临床治疗研究。结果①临床特点：在55例确诊慢性咳嗽的患儿中,以慢性顽固性咳嗽为主要症状。58％（32／55）的病例无肺部体征;②外周血：85％（47／55）的病例外周血变化不大,WBC（4—10）×10 9／L之间,嗜酸性粒细胞增多;③特别检查：47．27％（26／55）肺炎支原体IgM（MP—IgM）抗体阳性,83．64％（46／55）PeR技术检测肺炎支原体特异性DNA;④X光报告为多种形式。结论肺炎支原体（MP）感染是引起儿童慢性顽固性咳嗽的病因之一,对儿童慢性咳嗽,特别是顽固性咳嗽的诊治中应更加重视。     

Acetaminophen and acetylcysteine dose and duration: Past,present and future

Abstract

Acetylcysteine has been utilized successfully in the treatment of acetaminophen overdose since the 1970s. Although prospective trials as to efficacy and safety of acetylcysteine were conducted, there were no randomized controlled trials. This commentary addresses the reasons for this, and the background to choice of dose of acetylcysteine utilized in the oral and IV dosing regimens. Nomograms to predict possible hepatotoxicity based upon time of ingestion of acetaminophen were developed from a relatively arbitrary definition of toxicity as an aspartate aminotransferase/alanine aminotransferase (ALT/AST) greater than 1000 IU/L. While these have proved generally useful, patients still continue to develop hepatic damage after acetaminophen overdose, particularly if they present late after ingestion. The optimum management of these patients remains unclear, and one area of uncertainty is the dose and duration of acetylcysteine in various circumstances. This article discusses the issues that need to be elucidated to better target changes in acetylcysteine dose. The potential for measurements of other markers to improve treatment selection is the subject of further research.     

VEGF和NSE在良恶性嗜铬细胞瘤中的表达及意义

目的探讨肿瘤标志物血管内皮生长因子（VEGF）和神经元特异性烯醇化酶（NSE）在良、恶性嗜铬细胞瘤组织中的表达,分析其可能的临床价值及病理学意义,为临床鉴别良、恶性嗜铬细胞瘤提供辅助依据。方法应用免疫组化（SP法）检测16例恶性嗜铬细胞瘤、18例良性嗜铬细胞瘤及17例正常肾上腺髓质组织中细胞因子VEGF和NSE表达情况,显微镜下判断组织切片的染色结果。结果①恶性嗜铬细胞瘤VEGF表达明显强于正常肾上腺髓质和良性嗜铬细胞瘤（P〈0.01）。良性肿瘤和正常肾上腺髓质的VEGF表达差异无统计学意义（P〉0.05）。恶性嗜铬细胞瘤强阳性率明显高于良性嗜铬细胞瘤（P〈0.01）。②良、恶性嗜铬细胞瘤NSE表达差异有统计学意义（P〈0.05）,良性嗜铬细胞瘤NSE的表达高于正常肾上腺髓质的NSE表达（P〈0.05）。恶性嗜铬细胞瘤强阳性率高于良性嗜铬细胞瘤（P〈0.05）。③VEGF和NSE共同阳性表达在良、恶性嗜铬细胞瘤之间差异有统计学意义（P=〈0.01）。结论临床上检测VEGF和NSE可能为鉴别良、恶性嗜铬细胞瘤提供辅助依据,共同检测VEGF和NSE可能提高良、恶性嗜铬细胞瘤鉴别的敏感性。