微量酶联夹心杂交法定量检测hIL-8 mRNA PCR扩增产物

目的:建立一种灵敏度高、特异性强、定量、精确、操作简便的微孔板酶联夹心杂交技术,定量检测人IL-8 mR-NA。方法:针对人IL-8 mRNA逆转录聚合酶链反应产物一条链的不同区域序列设计一对特异探针,其中一条为捕获探针,5′端用活性氨基修饰,与微量DNA结合板表面的NOS基团共价结合,“竖直”地包被在微孔板内;另一条检测探针的3′端标记生物素,和辣根过氧化物酶结合。提取人外周血单个核细胞总RNA,进行RT-PCR扩增IL-8 mRNA,双链DNA产物经热变性后加入已包被捕获探针的微孔板内进行杂交,加入检测探针与已杂交的产物结合,经亲和素-辣根过氧化物酶系统检测杂交信号。结果:该法灵敏度为检出16个循环的PCR产物、5×103个PBMCs中的IL-8 mRNA、检测PCR终产物的最高稀释倍数为1∶256阳性;特异性试验非目的扩增片段酶联杂交检测未检出阳性杂交信号;精密度试验CV为5.2%。结论:该方法具有操作简便、灵敏度高、特异性强等优点,适合IL-8 mRNA PCR扩增产物的定量检测。     

Shared allergenic and antigenic determinants in Alternaria and Stemphylium extracts

To develop a model for mold allergen extract standardization, we studied eight commercial Alternaria extracts from various suppliers by a variety of immunochemical and physicochemical techniques, including measurement of Alt-I, a purified allergenic fraction of Alternaria. Wide variations were noted in the allergenic and antigenic potencies of these extracts. Estimates of Alt-I content measured by Alt-I RAST inhibition and by radioimmunoassay correlated significantly (p < 0.05), but Alt-I activity by either method could not be correlated with allergenic potency as measured by RAST inhibition using solid-phase Alternaria. Each test extract produced unique and differing patterns of Coomassie blue-stained bands in isoelectrofocusing gels and in crossed immunoelectrophoresis gels using rabbit antibodies to Alternaria. The optimal method for mold allergen standardization involves a combination of RAST inhibition, isoelectrofocusing, and crossed immunoelectrophoresis techniques, and, if possible, quantitation of individual allergens.     

IL-4–BATF signaling directly modulates IL-9 producing mucosal mast cell (MMC9) function in experimental food allergy

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Developmental regulation of type 1 and type 3 interferon production and risk for infant infections and asthma development

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Synergistic convergence of microbiota-specific systemic IgG and secretory IgA

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骨的结构与表征参数

阐述了骨的微米结构、纳米结构、表征参数与测量方法、及研究进展。     

Analysis of the α/β T-cell receptor repertoire by competitive and quantitative family-specific PCR with exogenous standards and high resolution fluorescence based CDR3 size imaging

The characterization of the human T-cell receptor (TCR) repertoire in various physiological and pathological conditions has become an important tool in studies of the immune response. Therefore, a number of PCR based strategies for the semiquantitative analysis of the TCR repertoire have been described. Family specific amplification of TCR cDNA has been employed in a number of studies often with contradictory results. We have developed a strategy utilizing exogenous standards with homologous primer binding sites for the quantitative analysis of the α/β T-cell receptor repertoire. This system allows the detection of even minute differences in T-cell populations based on quantitative PCR (Q-PCR) and competitive PCR (C-PCR). Results presented here demonstrate that expansions of T-cell subsets as defined by the specificity of the variable gene segments can be readily monitored when exceeding 1% of the total repertoire. In addition, the proposed method reveals direct information of CDR3 size heterogeneity and can be used to estimate the T-cell repertoire complexity and monitor clonal expansions. We discuss variables such as cell number and experimental conditions influencing accuracy and reproducibility of the analyses. We have used this protocol based on non-radioactive techniques for characterization of the fine specificity of the T-cell repertoire in peripheral and organ-infiltrating T-lymphocytes. The analyses revealed information about polyclonal or clonal expansion of T-cells in vivo and in vitro following various stimuli such as superantigenic stimulation of T-cell subsets as well as antigen-driven shaping of the α/β T-cell repertoire in autoimmune and infectious diseases.     

Increased fluidity of Plasmodium berghei-infected mouse red blood cell membranes detected by electron spin resonance spectroscopy

The fluidity of Plasmodium berghei-infected mouse red cell membranes is increased over that of uninfected cells at both 24°C and 37°C. This was demonstrated by electron spin resonance spectroscopy using the hydrocarbon spin labels 2-dodecyl-2′,5,5′-trimethyloxazolidine-N-oxyl and 2-heptyl-2′ -hexyl-5,5′-dimethyloxazolidine-N-oxyl to label regions of the bilayer near its surface, and deeper within the hydrocarbon region, respectively. Arrhenius plots of the ‘empirical motion parameter’ (R_i) obtained from 2-heptyl-2′-hexyl-5,5′-dimethyloxazolidine-N-oxyl-labeled cells versus temperature over the range from 0 to 45°C showed an hysteretic behavior of the spin labels in the membranes of both mature and immature uninfected cells. Such hysteretic behavior was consistently lacking in membranes of infected cells. These differences in membrane fluidity and spin label behavior are interpreted to reflect biochemical modifications of the red cell membrane which occur with infection by the malarial parasite.     

时间治疗学的计量研究

以《中国生物医学文献光盘数据库》(CBM)、《中文生物医学期刊文献数据库》(CMCC)和 OVID为数据源 ,全面检索时间治疗学的文献信息 ,利用文献管理软件 Pro Cite5辅助以手工方法对获取的信息进行管理和计量分析 ,以纳入文献的年代、期刊、著者、主题、国别和机构分布等科学和文献计量学指标全面揭示和评价时间治疗学的研究现状和发展趋势。研究结果显示 :时间治疗学的中文文献 91篇 ,分布于 73种期刊中 ,主题分布于中医学、心血管系统疾病、肿瘤、哮喘、消化性溃疡、糖尿病和时间治疗学总论等方面 ;同时纳入来自 35个国家和地区的有关外文文献 4 80篇 ,主要数据来源于 EMBASE和 MEDL INE,分布于 2 85种期刊中 ,收录时间治疗学文献 5篇以上的期刊有 14种 ,著者包括 12位 ,收录时间治疗学文献前 11名的机构分别为鲍尔勃罗斯医院、德州大学、康涅狄格州医学院、日本自治医学院、明尼苏达大学等。有关时间治疗学的研究 ,国内尚处于临床应用初期 ,国外已经形成核心著者、核心期刊和核心研究机构 ,主要分布在欧美等发达国家 ,已经在多个方面取得成绩。     

前列腺液白细胞含量与男性不育的相关性分析

目的探讨前列腺液白细胞含量与精液主要参数和指标的关系。方法对入选的260例男性不育患者进行前列腺液（EPS）常规检查计数白细胞和解脲支原体培养,按照WHO人类精液实验室手册要求检测精液主要参数、精子形态分析、精子顶体酶活性、精浆抗体（AsAb）等,分析前列腺白细胞与男性不育相关因素的关系。结果260例中炎症组93例（EPS中WBC≥10个/HP）：非炎症组（EPS中WBC〈10个/HP）167例。炎症组精液的精子密度、精子活动率、a＋b级活力精子率、精子顶体酶阳性率均低于非炎症组（P〈0.05）;而精液液化时间、精子畸形率、精浆抗体（AsAb）、前列腺液解脲支原体阳性率高于非炎症组（P〈0.05）。两组的精液量及pH值差异无显著性。结论前列腺液中白细胞含量对精液质量有一定的影响,慢性前列腺炎是导致男性不育的原因之一。