HLA-DRB1 allele polymorphisms in genetic susceptibility to esophageal carcinoma

AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal carcinoma in Han Chinese in Hubei Province. METHODS: HLA-DRB1 allele polymorphisms were typed by polymerase chain reaction with sequence-specific primers (PCR-SSP) in 42 unrelated patients with esophageal cancer and 136 unrelated normal control subjects and the associated HLA-DRB1 allele was measured by nucleotide sequence analysis with PCR.SAS software was used in statistics. RESULTS: Allele frequency (AF) of HLA-DRB1*0901 was significantly higher in esophageal carcinoma patients than that in the normal controls (0.2500 vs0.1397, P=0.028, the odds ratio 2.053, etiologic fraction 0.1282). After analyzed the allele nucleotide sequence of HLA-DRB1*0901 which approachs to the corresponded exon 2 sequence of the allele in genebank. There was no association between patients and controls in the rested HLA-DRB1 alleles. CONCLUSION: HLA-DRB1*0901 allele is more common in the patients with esophageal carcinoma than in the healthy controls, which is positively associated with the patients of Hubei Han Chinese. Individuals carrying HLA-DRB1*0901 may be susceptible to esophageal carcinoma.

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。

Construction and evaluation of DNA vaccines encoding Edwardsiella tarda antigens

Edwardsiella tarda is an opportunistic pathogen that can infect humans, animal, and fish. Two E. tarda antigens, Eta6 and FliC, which are homologues to an ecotin precursor and the FliC flagellin, respectively, were identified by in vivo-induced antigen technology from a pathogenic E. tarda strain isolated from diseased fish. When used as a subunit vaccine, purified recombinant Eta6 was moderately protective against lethal challenge of E. tarda in a Japanese flounder model, whereas purified recombinant FliC showed no apparent immunciprotectivity. Similarly, DNA vaccines based on eta6 and fliC in the form of plasmids pEta6 and pFliC induced, respectively, moderate and marginal protection against E. tarda infection. To improve the vaccine efficacy of eta6, a chimeric DNA vaccine, pCE6, was constructed, which encodes Eta6 fused in-frame to FliC. pCE6 was found to induce significantly higher level of protection than pEta6. Likewise, another chimeric DNA vaccine, pCE18, which expresses FliC fused to a previously identified E. tarda antigen Et18, elicited significantly stronger protective immunity than the DNA vaccine based on et18 alone. Fish immunized with pEta6 and pCE6 produced specific serum antibodies and exhibited significantly enhanced expression of the genes encoding elements that are involved in both innate and adaptive immune responses. Furthermore, the induction magnitudes of most of these genes were significantly higher in pCE6-vaccinated fish than in pEta6-vaccinated fish. (C) 2009 Elsevier Ltd. All rights reserved.

PROFIBUS DP从站协议栈开发与测试

本文研究了PROFIBUS DP欧洲标准EN50170的协议参考模型,并据此开发了ROFIBUSDP从站通信协议栈,最后采用荷兰PROCENTEC公司的ProfiScript工具进行了测试,本文给出了详细的软件编写和测试方法。

PROFIBUS-DP从站通信控制器的设计与实现

PROFIBUS是一种国际化、开放式、不依赖于设备生产厂商的现场总线标准，PROFIBUS-DP作为PROFIBUS的一个分支，以其成熟性、实时性、可靠性和稳定性，在全球范围内的工业自动化领域获得了最为广泛的应用。PROFIBUS-DP协议比较复杂, 目前只有少数国外厂商提供专用的PROFIBUS-DP协议芯片，而国内对于PROFIBUS-DP总线的应用基本以购买国外自动化设备厂商的PROFIBUS-DP通信芯片为主，导致我国的自动化行业难以掌握核心技术。因此研究和开发具有自主知识产权的PROFIBUS-DP通信芯片具有广阔的前景和重要的意义。本文通过深入研究PROFIBUS-DP协议，提出了一套完整的设计方案，并设计出符合PROFIBUS-DP协议的IP核，为最终PROFIBUS-DP通信芯片的实现打下了坚实的基础。 本文详细的介绍了PROFIBUS-DP从站通信控制器的设计实现过程。首先通过分析PROFIBUS-DP协议以及参考国外现有的芯片资料，结合自身研究，提出了PROFIBUS-DP从站通信控制器的整体设计方案，给出了设计的整体框图；其次在整体设计方案的基础上详细介绍了各个功能模块的实现方法，以此为基础，采用自顶向下的设计方法，对各个模块进行详细的设计，并给出了Verilog语言实现RTL编码以及核心功能模块的仿真波形图；最后采用ALTERA公司的Cyclone EP1C6 的FPGA芯片和Philips公司的P89LV51RD2 MCU搭建了一个标准化的智能型从站，并采用ProfiCore和ProfiScrit搭建了PROFIBUS-DP从站控制器的系统级验证环境，进行了系统级验证，充分证实了设计方案的可行性。

CD4+ CD25+ regulatory T cells control T helper cell type 1 responses to foreign antigens induced by mature dendritic cells in vivo.

Recent evidence suggests that in addition to their well known stimulatory properties, dendritic cells (DCs) may play a major role in peripheral tolerance. It is still unclear whether a distinct subtype or activation status of DC exists that promotes the differentiation of suppressor rather than effector T cells from naive precursors. In this work, we tested whether the naturally occurring CD4+ CD25+ regulatory T cells (Treg) may control immune responses induced by DCs in vivo. We characterized the immune response induced by adoptive transfer of antigen-pulsed mature DCs into mice depleted or not of CD25+ cells. We found that the development of major histocompatibility complex class I and II-restricted interferon gamma-producing cells was consistently enhanced in the absence of Treg. By contrast, T helper cell (Th)2 priming was down-regulated in the same conditions. This regulation was independent of interleukin 10 production by DCs. Of note, splenic DCs incubated in vitro with Toll-like receptor ligands (lipopolysaccharide or CpG) activated immune responses that remained sensitive to Treg function. Our data further show that mature DCs induced higher cytotoxic activity in CD25-depleted recipients as compared with untreated hosts. We conclude that Treg naturally exert a negative feedback mechanism on Th1-type responses induced by mature DCs in vivo.

Identification of major factors influencing ELISpot-based monitoring of cellular responses to antigens from Mycobacterium tuberculosis.

A number of different interferon-gamma ELISpot protocols are in use in laboratories studying antigen-specific immune responses. It is therefore unclear how results from different assays compare, and what factors most significantly influence assay outcome. One such difference is that some laboratories use a short in vitro stimulation period of cells before they are transferred to the ELISpot plate; this is commonly done in the case of frozen cells, in order to enhance assay sensitivity. Other differences that may be significant include antibody coating of plates, the use of media with or without serum, the serum source and the number of cells added to the wells. The aim of this paper was to identify which components of the different ELISpot protocols influenced assay sensitivity and inter-laboratory variation. Four laboratories provided protocols for quantifying numbers of interferon-gamma spot forming cells in human peripheral blood mononuclear cells stimulated with Mycobacterium tuberculosis derived antigens. The differences in the protocols were compared directly. We found that several sources of variation in assay protocols can be eliminated, for example by avoiding serum supplementation and using AIM-V serum free medium. In addition, the number of cells added to ELISpot wells should also be standardised. Importantly, delays in peripheral blood mononuclear cell processing before stimulation had a marked effect on the number of detectable spot forming cells; processing delay thus should be minimised as well as standardised. Finally, a pre-stimulation culture period improved the sensitivity of the assay, however this effect may be both antigen and donor dependent. In conclusion, small differences in ELISpot protocols in routine use can affect the results obtained and care should be given to conditions selected for use in a given study. A pre-stimulation step may improve the sensitivity of the assay, particularly when cells have been previously frozen.

Molecular basis of complete C4 deficiency. A study of three patients.

The highly polymorphic fourth component of human complement (C4) is usually encoded by two genes, C4A and C4B, adjacent to the 21-hydroxylase (21-OH) genes and is also remarkable by the high frequency of the null alleles, C4A*Q0 and C4B*Q0. Complete C4 deficiency is exceptional because this condition appears only in homozygotes for the very rare double-null haplotype C4AQ0,BQ0. This condition in most cases gives rise to systemic lupus erythematosus and an increased susceptibility to infections. The molecular basis for complete C4 deficiency has not yet been established. Therefore we studied the DNA of three previously described C4 deficient patients belonging to unrelated families by restriction fragment length polymorphism analysis using C4 and 21-OH probes. These studies revealed a deletion of the C4B and 21-OHA genes in two patients and no deletion at all in the third patient. Therefore, complete C4 deficiency as a result of homozygosity for the C4AQ0, BQ0 haplotype is not a consequence of a deletion of the C4 genes. The molecular basis of this genetic abnormality is certainly very complex and may vary also from one case to another.

Immunization with cocktail of HIV-derived peptides in montanide ISA-51 is immunogenic, but causes sterile abscesses and unacceptable reactogenicity.

BACKGROUND: A peptide vaccine was produced containing B and T cell epitopes from the V3 and C4 Envelope domains of 4 subtype B HIV-1 isolates (MN, RF, CanO, & Ev91). The peptide mixture was formulated as an emulsion in incomplete Freund's adjuvant (IFA). METHODS: Low-risk, healthy adult subjects were enrolled in a randomized, placebo-controlled dose-escalation study, and selected using criteria specifying that 50% in each study group would be HLA-B7+. Immunizations were scheduled at 0, 1, and 6 months using a total peptide dose of 1 or 4 mg. Adaptive immune responses in16 vaccine recipients and two placebo recipients after the 2nd immunization were evaluated using neutralization assays of sera, as well as ELISpot and ICS assays of cryopreserved PBMCs to assess CD4 and CD8 T-cell responses. In addition, (51)Cr release assays were performed on fresh PBMCs following 14-day stimulation with individual vaccine peptide antigens. RESULTS: 24 subjects were enrolled; 18 completed 2 injections. The study was prematurely terminated because 4 vaccinees developed prolonged pain and sterile abscess formation at the injection site-2 after dose 1, and 2 after dose 2. Two other subjects experienced severe systemic reactions consisting of headache, chills, nausea, and myalgia. Both reactions occurred after the second 4 mg dose. The immunogenicity assessments showed that 6/8 vaccinees at each dose level had detectable MN-specific neutralizing (NT) activity, and 2/7 HLA-B7+ vaccinees had classical CD8 CTL activity detected. However, using both ELISpot and ICS, 8/16 vaccinees (5/7 HLA-B7+) and 0/2 controls had detectable vaccine-specific CD8 T-cell responses. Subjects with moderate or severe systemic or local reactions tended to have more frequent T cell responses and higher antibody responses than those with mild or no reactions. CONCLUSIONS: The severity of local responses related to the formulation of these four peptides in IFA is clinically unacceptable for continued development. Both HIV-specific antibody and T cell responses were induced and the magnitude of response correlated with the severity of local and systemic reactions. If potent adjuvants are necessary for subunit vaccines to induce broad and durable immune responses, careful, incremental clinical evaluation is warranted to minimize the risk of adverse events. TRIAL REGISTRATION: ClinicalTrials.gov NCT00000886.