多发性硬化患者外周血单个核细胞SP3基因mRNA表达缺失研究

目的 研究我国多发性硬化(multiple sclerosis,Ms)患者外周血单个核细胞基因SP3(specificprotein 3)mRNA表达缺失的特点及其与临床表型的关系.方法 采集56例MS患者外周血单个核细胞,提取总RNA,设计SP3特异性引物,采用逆转录PCR方法观察SP3基因mRNA表达情况,并与其他疾病组、健康对照组进行比较.结果 56例MS患者中23例SP3为阴性结果,SP3表达缺失率为41.1%;健康对照组35例有5例为阴性结果,缺失率为8.6%;其它疾病对照组27例有4例为阴性结果,缺失率为14.3%.MS组与两对照组之间的差异有统计学意义(P＜0.01).SP3表达缺失组与表达组急性期的改良残废程度量表评分差异无统计学意义,稳定期则差异有统计学意义(P＜0.05).结论 通过对MS患者SP3基因mRNA表达的研究,观察到中国人MS存在该基因mRNA表达缺失.SP3与MS临床表现及其免疫学发病机制之间可能存在一定的相关性.     

多发性硬化与遗传研究进展

多发性硬化与遗传因素关系的研究受到极大重视 ,关联基因多态性与 MS易感性及病情程度的研究获得了一些进展 ,连锁分析的结果也支持 MS是一种遗传性疾病 ,本文总结有关这两方面的研究进展     

多发性硬化的免疫研究进展

多发性硬化(MS)是中枢神经系统(CNS)自身免疫性疾病的代表.本文对多发性硬化的免疫发病机制和免疫干预治疗的研究进展作一概述.     

人疱疹病毒-6与多发性硬化发病关联性研究

多发性硬化（MS）是典型的中枢神经系统炎症性自身免疫性疾病。很可能是青壮年神经系统失能的主要原因之一。作为一个复杂性状的疾病，多发性硬化可能是由环境因素和易感性基因相互作用而引起的一系列反应。包括免疫系统反应、轴突和神经胶质的急性炎症反应、功能恢复和结构的修复、炎症后神经胶质的增生以及神经节变性。     

多发性硬化的诱发电位分析

本对26例多发性硬化的病人进行脑干听觉诱发电位、视觉诱发电位,体感诱发电位的测定。以观察三种诱发电位在多发性硬化病人中的改变。经分析发现,三种诱发电位在多发性硬化病人中异常率较高,可提供第二个或多发的亚临床的客观证据,对本病的早期诊断有辅助价值,且对了解病情和预后有一定帮助。     

多发性硬化患者外周血淋巴细胞CD56的表达

目的 探讨CD56^ 淋巴细胞在多发性硬化（MS）的变化。方法 流式细胞仪测定32例复发缓解型MS患者（复发发期23例,缓解期9例）及13例复发期MS患者予糖皮质激素治疗后外周血淋巴细胞CD56表达的阳性百分率。结果 复发期和缓解期MS患者CD56的表达均高于对照组,15例复发期MS患者CD56的阳性百分率与血脑屏障受损呈正相关,复发期MS患者CD56的水平与距发作时间、整个病程、EDSS伤残评分无关。缓解期MS CD56的水平与病程无关,激素对CD56的表达我影响。结论 CD56^ 淋巴细胞涉及MS的发病机制。     

诱发电位对多发性硬化的诊断价值

本文报告多发性硬化(MS)112例,全组包括临床确诊79例,近似确诊33例。不正常的VEP占86.2％,SEP占80％,BAEP占66.7％。本组临床特点发病开始更常发生视觉障碍,以视神经损害为主。VEP可以发现无临床症状体征的视神经病灶,如果BAEP不正常提示脑干处有一临床下的病灶。因此,诱发电位检查可提供第二或多灶的病变,有助于对多发性硬化的早期诊断。     

多发性硬化的发病机制

多发性硬化是中枢神经系统炎性脱髓鞘疾病,有遗传易感性,与环境因素有关,其病因和发病机制尚未阐明,近年的研究表明它具有多种发病机制,并依此提出许多新论点和观念,我们对此进行了综述。     

寻找差异表达基因

在现代医学及分子生物学研究中 ,证实、分离、鉴定差异表达基因有着极其广泛的应用。发现差异表达基因 ,有助于阐明疾病发生的分子机制、临床诊断及预后的预测 ;并是基因药物和基因治疗发展的基础。然而 ,当前克隆、鉴定差异表达基因的技术中 ,仅少数几种技术利用了人类及模式生物基因组研究业已取得的成就。其中 ,基因表达的系列分析 (serialanaly sisofgeneexpression ,SAGE)、cDNA微点阵 (cDNAmicroarrays)和综合性基因鉴定程序 (integratedprocedureforgeneidentification ,IP GI)等方法将会变得更为有利。本文将简述近年来用于寻找差异表达基因的几种主要技术 ,并对其趋势作一展望。     

差异表达基因克隆技术的新进展

分离并克隆差异表达基因是目前功能性基因研究的热点。mRNA差异展示是筛选差异表达基因是有效的技术之一，它的主要不足是有一定假阳性。RDA、SSH、DSD、LSH技术能够克旺假阳性，但也具有一定的不足，应根据需要选择运用。随着能扩增长片段及具自动校正功能的Taq酶出现及应用，这些技术将会不断完善与发展，具有广阔的应用前景。     

多发性硬化患者外周血白细胞粘附分子的表达

目的 :探讨多发性硬化 (MS)患者外周血白细胞粘附分子表达的水平及予甲基强的松龙 (MP)治疗后的变化。方法 :流式细胞仪测定 2 8例缓解复发型MS患者及 12例复发期MS予静脉MP治疗后外围血淋巴细胞和单核细胞细胞间粘附分子 3(CD5 0 )、细胞间粘附分子 1(CD5 4 )、整合素LFA 1β亚单位 (CD18)、非常晚抗原 4 (VLA 4 )α、β亚单位 (CD4 9d、CD2 9)和 (- )选择素 (CD6 2L)的阳性百分率。结果 :复发期MSCD4 9d和CD2 9在淋巴细胞和单核细胞、CD5 4和CD6 2L在单核细胞的阳性百分率高于缓解期MS和对照组 ,复发和缓解期MSCD5 4在淋巴细胞的阳性百分率高于对照组 ;MP治疗后 ,CD5 4和CD4 9d在淋巴细胞和单核细胞、CD6 2L在单核细胞的阳性百分率下降。结论 :MS患者外周血白细胞粘附分子的表达升高并可作为MS活动期的指标     

多发性硬化患者外周血记忆性T细胞亚群的研究

目的：近来研究表明中心记忆性和效应记忆性T细胞构成记忆性T细胞的两个亚群，但在多发性硬化（Multiple sclerosis，MS）中的作用尚不清楚，因此测定了MS患者外周血记忆性T细胞亚群的水平，并进一步探讨了可能导致记忆性T细胞亚群变化的机制。方法：采用流式细胞仪和酶联免疫吸附分析（Enzyme-linked immunosorbent assay，ELISA）分别检测未治疗MS、正常对照组外周血记忆性T细胞亚群的阳性率和血浆IL-15浓度。进一步根据MS临床表现分为复发-缓解型MS（Relapse-remission MS，RRMS）和慢性进展型MS（Chronic progressiveMS，CPMS）两个亚组。结果：与正常对照比较，在CD8^＋T细胞亚群中，MS患者中心记忆性T细胞（Central memory T cell，TCM）、终末效应记忆性T细胞明显增多和减少（P〈0．05和P〈0．01），RRMS患者终末效应记忆性T细胞明显少于正常对照（P〈0．05）；MS患者血浆IL-15水平较正常对照明显升高（P〈0．05）。结论：研究显示了Ms患者中CD8^＋T CM上调，可能反映了在MS早期被诱导产生的一个持续慢性炎性应答，我们推测CD8^＋T CM的异常改变可能在维持MS慢性炎症的过程中起着重要作用，而IL-15则可能参与了促进TCM分化的调节过程。     

应用mRNA差异显示技术筛选大鼠小脑颗粒神经元凋亡模型中差异表达基因

目的： 应用mRNA差异显示技术对大鼠小脑颗粒神经元凋亡模型中差异表达基因进行筛选，克隆以及鉴定。方法: 大鼠小脑颗粒神经元的分离和原代培养；对大鼠小脑颗粒神经元凋亡逆转模型进行荧光mRNA差异显示逆转录PCR（FDDRT-PCR,fluorescent differential display RT-PCR）；EST片段亚克隆入pGEM-T EasyTM，克隆后测序并进行序列同源性检索；反Northern杂交重鉴定与筛选。结果: 通过DDRT-PCR获得164 个在小脑颗粒神经元凋亡模型中差异表达的ESTs；对其中的17个ESTs进行了克隆并测序；通过反Northern杂交的初步筛选初步鉴定出5个阳性片段。结论: 阳性差异表达EST的筛选、克隆以及鉴定为神经元凋亡与保护分子机制研究奠定基础。     

人外周血细胞放射损伤相关基因的生物信息学

目的从基因水平揭示放射前后人外周血细胞的变化。方法从基因表达数据库(GEO)中下载两组经放射处理后人外周血基因芯片数据,利用Qlucore Omics Explorer 3.0软件筛选差异表达基因,STRING、DAVID等在线分析工具对差异表达基因进行下一步的生物信息学分析。结果共筛选出94个共同差异表达基因,其中共同表达上调31个,共同表达下调11个,这些差异表达基因主要涉及到细胞凋亡的调控,细胞程序性死亡的调控,细胞死亡的调控,细胞周期的调控,DNA损伤应答,胞内信号转导等的分子功能及生物学过程。通过STRING分析,发现泛素C(UBC)、增殖细胞核抗原(PCNA)、鼠双微体基因-2(MDM2)处在核心节点位置,并参与p53通路。结论通过生物信息学的方法分析得出UBC、PCNA、MDM2可能成为潜在的防护放射后损伤的靶点。     

Assessment of expression of RELN signaling pathway in multiple sclerosis patients

Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system. Nearly 85% of MS patients are recognized with relapsing-remitting MS (RRMS), a typical clinical course of disease which is distinguished by several episodes of relapses, separated by remissions of neurological impairment. Failure of repair mechanisms is a main factor in progression of neurological dysfunction in MS. Several lines of evidence suggest that Reelin (RELN) signaling pathway can contribute in the regulation of repair mechanisms in MS patients. In the present study, we assessed expression levels of RELN and Disabled-1 (DAB1), two key genes in RELN signaling pathway, in peripheral blood of 50 RRMS patients and 50 matched healthy subjects. RELN was significantly down-regulated in total MS patients, and total female patients compared with the matched controls. However, no statistically significant difference was found in DAB1 mRNA expression between MS patients and controls. Furthermore, considerable correlations were detected between expression levels of RELN and DAB1 in the patients group. There were no significant correlations between expression levels of genes and EDSS, disease duration or age at onset. Our study provides evidences for the role of RELN signaling pathway in the pathogenesis of MS. Further studies are required to clarify the exact clinical significance of this pathway in MS patients.     

Red blood cell glutathione peroxidase activity in multiple sclerosis

Summary Glutathione peroxidase (GSH-Px) activity of red blood cells of 23 multiple sclerosis (MS) patients (10 men and 13 women, aged 22–64 years) was examined and compared to the enzyme activity of 26 healthy persons (15 men and 11 women, aged 19–50 years). It was found that the mean GSH-Px activity was significantly higher (P<0.001) in red blood cells of MS patients (39.1±8.1 IU/g Hb) as compared to the group of healthy persons (25.9±5.2 IU/g Hb). There was no difference according to sexes in both the MS patients and the control group. The results are discussed based on the hypothesis that organic peroxides play a role in the etiology of multiple sclerosis.     

Increased blood vessel density and endothelial cell proliferation in multiple sclerosis cerebral white matter

Multiple sclerosis (MS) is primarily considered an inflammatory demyelinating disease, however the role of vasculature in MS pathogenesis is now receiving much interest. MS lesions often develop along blood vessels and alterations in blood brain barrier structure and function, with associated changes in the basement membrane, are pathological features. Nevertheless, the possibility of angiogenesis occurring in MS has received little attention. In this study we used triple label enzyme immunohistochemistry to investigate blood vessel density and endothelial cell proliferation in MS samples (n = 39) compared with control tissue to explore evidence of angiogenesis in MS. The results showed that in all MS samples examined blood vessel density increased compared with controls. The greatest increase was found in subacute lesions where numbers of positively stained vessels increased from 43.9 ± 8.5% in controls to 84.2 ± 13.3% (P = 0.001). Furthermore, using an antibody against endoglin (CD105), a specific marker of proliferating endothelial cells, which are characteristic of angiogenesis, we have shown that vessels containing proliferating endothelial cells were more pronounced in all MS tissue examined (normal-appearing white matter, acute, subacute and chronic lesions, P ≥ 0.027) compared with control and this was greatest in the MS normal-appearing white matter (68.8 ± 19.8% versus 10.58 ± 6.4%, P = 0.003). These findings suggest that angiogenesis may play a role in lesion progression, failure of repair and scar formation.     

环孢素A受体在多发性硬化病人外周血单个核细胞中的表达

目的 通过对多发性硬化患者外周血单个核细胞中环孢素Ａ受体ｍＲＮＡ表达的研究，为临床采用环孢素Ａ辅助治疗该病提供一定的依据。方法 采用逆转录ＰＣＲ方法，结果经凝胶图像分析。结果 患者组即多发性硬化患者外周血单个细胞存在有ＣｙＰｍＲＮＡ的表达，与对照组相比较降低，尚无明显统计学差异（Ｐ〉０．０１）。结论 多发性硬化患者外周血单个核细胞中存在有ＣｙＰ，ＣｓＡ与细胞内的ＣｙＰ结合后是否产生生物活性还与其蛋     

Increased gene expression of scavenger receptors and proinflammatory markers in peripheral blood mononuclear cells of hyperlipidemic males

Interactions between peripheral blood mononuclear cells (PBMCs) and those within plaques are suggested to be pathophysiologically relevant to lipid-induced arteriosclerosis. In this study, gene expressions of scavenger receptors (CD36, CD68), LPS receptor (CD14), proinflammatory (tumor necrosis factor alpha [TNFalpha], CD40, interleukin-1 beta [IL-1beta]) and oxidative stress-related (manganese superoxide dismutase [MnSOD]) markers were analyzed in PBMCs of clinically asymptomatic males with classical proatherogenic risk factors such as smoking and/or hyperlipidemia. PBMCs were isolated from venous blood of normolipidemic non-smokers (n = 10) and smokers (n = 8), and hyperlipidemic non-smokers (n = 9) and smokers (n = 8). RNA from PBMCs was used for PCR analyses. Plasma concentrations of oxidized low-density lipoproteins (oxLDL) were measured by ELISA. The gene expressions of CD36, CD68, CD40, TNFalpha, and MnSOD were significantly higher in PBMCs of hyperlipidemics than in normolipidemics, irrespective of whether they were smoking or not. The individual expression of these genes showed significant positive correlations with each other but also with serum cholesterol or plasma oxLDL concentrations. The higher expressions of scavenger receptors, proinflammatory and oxidative stress-related genes of PBMCs are suggested to result mainly from hyperlipidemia and the accompanied increase of oxLDL concentrations.