湖北食管癌HLA-DQB1的基因多态性

目的从基因水平探讨湖北地区汉族人食管癌 HEN-DQB1等位基因的遗传易感性.方法运用序列特异性引物聚合酶链反应技术,检测无亲缘关系湖北汉族健康人136例、食管癌组42例患者的 HLA-DQB1等位基因.SAS system 统计软件数据处理.结果湖北汉族人食管癌患者与正常人比较,HEN-DQB1*0301基因频率显著增高(0.2976 vs 0.1875),P=0.046,OR=1.835,病因分数=0.1354);两组间 HLA-DQB1其余各等位基因分布频率的比较,HLA-DQB1*0201(0.0

Evaluation of Clonorchis sinensis recombinant 7-kilodalton antigen for serodiagnosis of clonorchiasis

The diagnostic applicability of the Clonorchis sinensis recombinant 7-kDa protein was evaluated. In enzyme-linked immunosorbent assays and immunoblots, the protein showed high sensitivities (81.3 and 71.9%, respectively) and specificities (92.6 and 89.7%, respectively) for sera obtained from various helminthic infections. Some paragonimiasis sera showed cross-reactions. The antigen might be valuable in the serodiagnosis of human clonorchiasis.

基于HLA卫星组网仿真想定与HLA Web化扩展的研究

卫星组网仿真是对各种卫星组网方案进行性能分析、效能评估以及优化设计的有效途径。限于单机负载及计算能力，卫星组网仿真采用分布式交互仿真形式。作为仿真的基本依据，想定编辑生成与想定发布是仿真首先需要解决的问题。此外，想定推演过程也应对仿真场景进行展示以方便观察仿真当前状态。如何构建满足需求的卫星组网仿真想定系统是本文研究目的之一。 围绕卫星组网仿真想定系统的功能需求，首先进行了需求分析与功能定义，接着对所涉及难点包括多任务多粒度组网仿真想定的灵活配置、细粒度组网仿真内存使用优化以及想定数据实时发布等的技术途径进行了比较与分析。在确定所选用技术基础上，设计出想定系统的总体结构框架。最后结合该框架对系统实现进行了详细的说明，并对内存优化问题进行了仿真实验。 在想定系统实现基础上，进一步分析了当前高层体系结构HLA标准在分布式式交互仿真应用中存有的缺陷，给出利用Web服务对其进行扩展的必要性和扩展途径。通过对面向服务的体系结构SOA及其实现技术Web服务的讨论，分析了通过Web服务扩展HLA使其Web使能的多种途径。文中提出一种基于代理、使用Web服务的HLA Web化扩展方法，并给出了设计方案。

一种基于HLA的卫星仿真系统的设计与实现

针对当前卫星仿真中仿真工具可扩展性差和空间环境模型复杂的问题,设计并实现了一种基于HLA的分布式卫星仿真系统。在架构设计层面,遵循分布式系统设计思想,把系统自底向上划分为数据支撑层、仿真支撑层和仿真应用层,并基于负载均衡的考虑,设计了四个功能集中的仿真应用子系统。在系统实现层面,结合虚拟现实技术,实现了想定方案的灵活配置以及星空环境和在轨卫星的实时渲染,搭建了支持多种卫星仿真应用的通用仿真运行平台。在仿真应用层面,实现了在此平台上对卫星编队变轨过程的仿真,并实时采集仿真数据对仿真结果进行分析和评估。

基于HLA的仿真模型管理方法研究

“仿真是一种基于模型的活动”，任何仿真系统都不能离开模型的支持，如果每次开发新的系统都要重新建立模型，费时费力。随着仿真系统的日益复杂，导致仿真模型的结构也日趋复杂，模型管理亦日趋繁琐。因此，研究一种有效的模型管理方法，对于方便模型重用，提高开发效率有着重要的意义。实现对模型的有效管理，首先需要明确管理对象，然后把模型有条理的分类并且规范的描述出来，最后把模型存储在数据库中，供用户重用。论文首先在HLA联邦开发执行过程的基础上，分析和完善了HLA仿真建模体系，明确了HLA仿真中模型的层次；然后总结了现有的模型分类方法，从方便模型统一管理的角度，提出了一种可扩展的模型分类方法；引入了元数据和XML技术，实现了对模型的规范化描述；根据课题研究目的，提出了仿真模型管理系统的设计目标，并设计了系统的体系结构、功能模块和数据结构；最后，综合应用数据库、VC++等技术，实现了模型存储、模型的增、删、改、查以及用户管理等功能，实现了对于模型的统一管理。

Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reactions

We used colloidal An to enhance the amount of antibody immobilized on a gold electrode and ultimately monitored the interaction of antigen-antibody by impedance measurement. Self-assembly of 6 nm (diameter) colloidal An onto the self-assembled monolayers (SAMs) of 4-aminothiophenol modified gold electrode resulted in an easier attachment of antibody. The redox reactions of [Fe(CN)(6)](4-)/[Fe(CN)(6)](3-) on the gold surface were blocked due to the procedures of self-assembly of 4-aminothiophenol and antibody immobilization, which were investigated by cyclic voltammetry and impedance spectroscopy. The interaction of antigen with grafted antibody recognition layers was carried out by soaking the modified electrode into a phosphate buffer at pH 7.4 with various concentrations of antigen at 37 degreesC for 30 min. The antibody recognition layers and their interactions with various concentrations of antigen could be detected by measurements of the impedance change. The results show that this method has good correlation for detection of Hepatitis B virus surface antigen in the range of 0.5-200 mug/l and a detection limit of about 50 ng/l.

Immunosensors based on layer-by-layer self-assembled Au colloidal electrode for the electrochemical detection of antigen

Colloidal Au particles have been deposited on the gold electrode through layer-by-layer self-assembly using cysteamine as cross-linkers. Self-assembly of colloidal Au on the gold electrode resulted in ail easier attachment of antibody, larger electrode surface and ideal electrode behavior. The redox reactions of [Fe(CN)(6)]-/[Fe(CN)(6)](3-) on the gold surface were blocked due to antibody immobilization, which were investigated by cyclic voltammetry and impedance spectroscopy. The interaction of antigen with grafted antibody recognition layers was carried out by soaking the modified electrode into a phosphate buffer at pH 7.0 with various concentrations of antigen at 37degreesC for 30 min. Further, an amplification strategy to use biotin conjugated antibody was introduced for improving the sensitivity of impedance measurements. Thus, the sensor based oil this immobilization method exhibits a large linear dynamic range, from 5 - 400 mug/L for detection of Human IgG. The detection limit is about 0.5 mug/L.

Use of nitrocellulose films for affinity-directed mass spectrometry for the analysis of antibody/antigen interactions

Combination of affinity extraction procedures with mass spectrometric analyses is termed affinity-directed mass spectrometry, a technique that has gained broad interest in immunology and is extended here with several improvements from methods used in previous studies. A monoclonal antibody was immobilized on a nitrocellulose (NC) membrane, allowing the corresponding antigen to be selectively captured from a complex solution for analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). This method was also used to rapidly determine the approximate binding region responsible for the antibody/antigen interaction. The tryptic fragments of antigen protein in buffer were applied to the antibody immobilized on NC film and allowed to interact. The NC film was then washed to remove salts and other unbound components, and subjected to analysis by MALDI-TOFMS. Using interferon-alpha (2a) and anti-interferon-alpha (2a) monoclonal antibody IgG as a model system, we successfully extracted the antigen protein and determined the approximate binding region for the antigen/antibody interaction (i.e., the tryptic fragment responsible). Copyright (C) 2001 John Wiley & Sons, Ltd.

Amplification of antigen-antibody interactions based on biotin labeled protein-streptavidin network complex using impedance spectroscopy

Antibody was covalently immobilized by amine coupling method to gold surfaces modified with a self-assembled monolayer of thioctic acid. The electrochemical measurements of cyclic voltammetry and impedance spectroscopy showed that the hexacyanoferrate redox reactions on the gold surface were blocked due to the procedures of self-assembly of thioctic acid and antibody immobilization. The binding of a specific antigen to antibody recognition layer could be detected by measurements of the impedance change. A new amplification strategy was introduced for improving the sensitivity of impedance measurements using biotin labeled protein- streptavidin network complex. This amplification strategy is based on the construction of a molecular complex between streptavidin and biotin labeled protein. This complex can be formed in a cross-linking network of molecules so that the amplification of response signal will be realized due to the big molecular size of complex. The results show that this amplification strategy causes dramatic improvement of the detection sensitivity of hIgG and has good correlation for detection of hIgG in the range of 2-10 mug/ml. (C) 2001 Elsevier Science B.V. All rights reserved.

STUDIES ON INCREASING THE ACTIVITY OF SELENIUM-CONTAINING ABZYME .1. SYNTHESIS, CHARACTERIZATION AND PROPERTIES OF HAPTEN AND ANTIGEN

The thiol group of glutathione (GSH) reacts specifically with 2,4-di-ni-trochlorobenzene to give S-substituted dinitrophenyl glutathione (GSH-S-DNP); two carboxyl groups of GSH-S-DNP were further esterified by n-butanol to produce the hapten, multisubstrate analog GSH-S-DNP Butyl Ester (GSH-S-DNP BE). The primary structure of the hapten was characterized by the free. amino group analysis, H-1 NMR, IR determinations and the elemental analysis. The hapten was then conjugated to bovine serum albumin (BSA) in the presence of glutaraldehyde. The reaction mixture was purified by Ultrogel AcA54 colum chromatography to give the antigen. On an average, 25 haptens were bound to each BSA molecule. Electrophoresis analysis showed that the average molecular weight of the antigen was 87 KD. CD spectrum showed that the a-helix content of the antigen increased.

Identification and immunoprotective analysis of an in vivo-induced Edwardsiella tarda antigen

Edwardsiella tarda is a severe aquaculture pathogen that can infect many important fish species cultured worldwide. The aim of this study was to evaluate the vaccine potential of an E. tarda antigen, Eta21, which was identified from a pathogenic E. tarda strain via the method of in vivo-induced antigen technology (IVIAT). Eta21 is 510-amino acid in length and shares similar to 58% sequence identity with a putative peptidase of several bacterial species. eta21 was subcloned into Escherichia colt, and recombinant Eta21 was purified as a histidine-tagged protein. When used as a subunit vaccine, purified recombinant Eta21 was effective against lethal E. tarda challenge in a Japanese flounder model. In order to improve the immunoprotective efficacy of Eta21, the chimera AgaV-Eta21 was constructed, which consists of Eta21 fused in-frame to the secretion domain of AgaV, an extracellular beta-agarase. E. coli DH5 alpha harboring plasmid pTAET21, which constitutively expresses agaV-eta21, was able to produce and secret AgaV-Eta21 into the extracellular milieu. Vaccination of Japanese flounder with live DH5 alpha/pTAET21 elicited immunoprotection that is significantly higher in level than that induced by vaccination with purified recombinant Eta21. Vaccination with DH5 alpha/pTAET21 and recombinant Eta21 both induced the production of specific serum antibodies at four to eight weeks post-vaccination. Taken together, these results demonstrate that Eta21, especially that delivered by DH5 alpha/pTAET21, is an effective vaccine candidate against E. tarda infection. (C) 2009 Elsevier Ltd. All rights reserved.