东部马脑炎病毒E2基因与GM-CSF共表达质粒的构建及其免疫原性初步研究

目的:利用粒细胞-巨噬细胞集落刺激因子(GM-CSF)作为基因佐剂,提高东部马脑炎病毒E2基因重组质粒的免疫效果.方法:分别扩增E2基因与GM-CSF基因,连接进入含有内部核糖体插入位点(IRES)的真核表达载体中,构建共表达质粒.Lipofectamine2000介导转染真核细胞BHK-21,反转录PCR检测E2和GM-CSF两个基因的转录,Western印迹法和间接免疫荧光法(IFA)检测细胞内E2基因表达产物的抗原性.通过肌肉注射免疫BALB/c小鼠,IFA法测定血清抗体效价,FACS测定免疫小鼠脾细胞中CD4 /CD8 淋巴细胞构成比,初步观察共表达质粒的免疫原性.结果:经酶切鉴定与序列测定证明重组共表达质粒中含有E2和GM-CSF两个基因且序列正确.转染细胞的反转录PCR可见E2和GM-CSF两个基因的转录.Western印迹结果可见转染细胞的裂解液在相对分子质量50 000位置有特异性条带.将转染细胞制成荧光抗原片,经IFA检测可见胞浆中出现特异荧光.免疫小鼠的最高血清抗体效价为1:160,但细胞免疫未观察到明显的提高.结论:构建的共表达质粒pE2-IRES-mGM-CSF具有良好的免疫原性,能有效刺激小鼠特异性体液免疫应答,提高了E2基因重组质粒诱导的血清抗体效价.     

抗晶状体蛋白的特异性抗体免疫示踪研究

目的：了解针对晶状体组织的特异性抗体到达眼前节的途经、部位及其对晶状体生理功能的影响。方法：采用荧光标记抗体，观察被动免疫动物眼前节组织中荧光反应的变化，测定晶状体调节力和进行眼前节裂隙灯检查。结果：特异性抗体(IgG)首先到达睫状体部，而后出现在晶状体囊膜、虹膜和角膜内皮上；眼前节抗体含量峰值与裂隙灯观察的房闪、即同时出现；晶状体上皮细胞的损伤与晶状体的调节力下降则出现于转移免疫后2w。结论：针对晶状体组织抗原的特异性抗体可通过体循环先到达睫状体部，然后随房水而作用于晶状体上皮细胞。     

Analysis of gene expression in MOG-induced experimental autoimmune encephalomyelitis after treatment with a novel brain-penetrating antioxidant

Accumulating data from experimental studies indicate that oxidative stress has a major role in the pathogenesis of multiple sclerosis (MS). It has been suggested that local production of reactive oxygen species, probably by macrophages, mediates axonal damage in both MS patients and the mouse model experimental autoimmune encephalomyelitis (EAE). We have shown previously that our novel brain-penetrating antioxidant, N-acetylcysteine amide (AD4), reduces the clinical and pathological symptoms, including inflammation and axonal damage in myelin oligodendrocyte glycoprotein (MOG)-induced chronic EAE in mice. The aim of this study was to examine the molecular mechanism by which AD4 exerts protection in MOG-induced EAE mice. Therefore, we analyzed gene-expression profile in the spinal cords of MOG-induced chronic EAE mice and compared them with MOG-induced mice treated with AD4, using a cDNA microarray. We found that MOG treatment up-regulated genes encoding growth factors, cytokines, death receptors, proteases, and myelin structure proteins, whereas MOG- and AD4-treated mice demonstrated gene expression profiles similar to that seen in na?ve healthy mice. In conclusion, our study shows that chronic AD4 administration suppresses the induction of various pathological pathways that play a role in EAE and probably in MS.     

产褥期脑病62例临床分析

目的研究产褥期脑病的临床特征、治疗方法及预后。方法选择62例产褥期突发精神症状并有全脑受累体征病人的临床资料进行回顾性分析。结果发现产褥期脑病的临床表现酷似病毒性脑炎,但病前无明显感染灶,病程较长,疗效和预后较差。结论对产褥期精神异常病人应进行神经病学方面检查,尽早诊治。病初要控制和缩短脑水肿期,防止和减轻脑细胞的变性和脱鞘;恢复期要耐心而持久地进行各种康复治疗,以便病人恢复到最佳健康水平。     

散发性脑炎频谱分析

目的探讨散发性脑炎患者的定量脑电图频谱分布特征及该技术临床应用的可能性。方法对４３例临床确诊的散发性脑炎和２０例健康对照组进行定量脑电图频谱分析。结果散发性脑炎组主频均数＜６Ｈｚ，与对照组比较，Ｐ＜０．０ｌ，散发性脑炎组σ、θ相对功率值均比对照组明显增高。结论定量脑电图主频分布及各频段相对功率值在诊断散发性脑炎中有一定的临床应用价值。     

Antibody Mediated Complete Congenital Heart Block in the Fetus

Complete congenital heart block (CCHB) affects 1:20,000–25,000 live births and is usually an atrioventricular block; 30–50% of fetuses with CCHB will have a structural anomaly, though recently attention has focused on the etiological influence of autoimmune disease, such as systemic lupus erythematosus. The diagnosis is established by detailed two-dimensional ultrasound scanning of the heart to exclude anomaly coupled with M-mode echocardiography and Doppler blood velocity patterns in the major vessels to detect the uncoupling of atrial and ventricular rhythms. Risk factors for an affected child are discussed. A previously affected child, high titers of anti-Ro antibodies, the presence of anti-Ro (SS-A) and anti-La (SS-B), and maternal HLA DR3 confer high risk. Antibody mediated CCHB is irreversible. Plasmapheresis and immunosuppression have been attempted in affected mothers, with limited success, to reduce the likelihood of the fetus being affected, and steroids have been used to reduce the inflammatory reaction in the heart. In many cases the underlying pathology of the immune system adversely affects utero-placental function requiring careful monitoring of fetal well-being. CCHB renders fetal heart rate monitoring virtually useless, in the antenatal and intrapartum periods. Alternatives are explored.     

Granulocyte-macrophage colony stimulating factor inhibits class II major histocompatibility complex expression and antigen presentation by microglia

Granulocyte-macrophage colony stimulating factor (GM-CSF) modulates various functions of monocytes/ macrophages including antigen-presenting capacity. Recently it was found that astrocytes produce GM-CSF in the central nervous system (CNS) and that GM-CSF can induce proliferation and morphological changes of microglia. Here we show that GM-CSF can down regulate the interferon-γ-mediated induction of major histocompatibility complex (MHC) class II antigens in microglia, but not in astrocytes. GM-CSF pretreatment completely prevents myelin basic protein-specific T cell proliferation induced by microglia but not astrocytes. GM-CSF did not affect the cell surface expression on microglia of either MHC class I or cell adhesion molecules. The inhibition of microglial MHC class II expression and antigen-presenting function is specific for GM-CSF, as treatment with a different CSF (interleukin-3) did not modulate microglial phenotype or functional capacity. These data suggest that GM-CSF might be involved in the regulation of immune responses within the central nervous system.     

Ache、P-selectin、CDK5在病毒性脑炎中的表达及相关性研究

目的 探究乙酰胆碱酯酶(acetylcholinesterase, Ache)、P-选择素(P-selectin)、细胞周期素依赖蛋白激酶5(cyclin-dependent kinase 5,CDK5)在病毒性脑炎(viral encephalitis, VE)中的表达及相关性。方法 本研究选取在本院健康体检及神经内科就诊治疗、住院治疗的病毒性脑炎患者130例作为研究对象,并纳入研究组,根据患者的病情时间将其分为急性期和恢复期,其中急性期组85例,恢复期组45例。根据患者的临床表现,将其分为轻度组和重度组,其中轻度组76例,重度组54例,选择同期在我院接受体检的健康患者60例作为对照组。应用丁酰硫代胆碱底物法检测Ache水平;酶联免疫吸附法检测血清P-selectin水平;CDK5分离试剂盒检测CDK5水平。观察研究组和对照组、急性期和恢复期及重度组和轻度组患者Ache、P-selectin、CDK5水平,分析病毒性脑炎患者Ache、P-selectin和CDK5之间相关性,利用ROC曲线比较Ache、P-selectin、CDK5单独检测及联合检测在VE中的诊断价值。结果 研究组A...