血清可溶性 CTLA-4表达与重症肌无力的相关性研究

目的探讨血清可溶性细胞毒性T淋巴细胞相关抗原-4（CTLA-4）表达与福建地区重症肌无力（ myasthenia gravis ,MG）患者的相关性。方法 ELISA法检测福建地区80例MG患者（ MG患者分为激素未治疗组及激素治疗组,且激素治疗组进一步分为免疫抑制剂治疗组和胸腺切除组）和80例正常对照组血清可溶性CTLA-4（ sCTLA-4）水平。结果 MG激素未治疗组、激素治疗组的sCTLA-4均高于正常对照组[（6．03±3．58） ng/ml、（3．44±2．36） ng/ml vs （0．49±0．95） ng/ml],它们之间的差别具有统计学意义（χ2＝100．67,P＜0．001）;其中激素治疗组sCTLA-4表达水平高于正常对照组（Z＝-9．03,P＜0．001）,而 MG激素未治疗组sCTLA-4表达高于激素治疗组（Z＝-3．37,P＝0．001）;并且激素未治疗组激素治疗前后血清sCTLA-4水平差异有统计学意义（t＝3．10,P＝0．005）;胸腺切除术后sCTLA-4低于手术前（Z＝-2．21,P＝0．04）,而免疫抑制剂治疗前后差异却没有统计学意义（Z＝-1．26,P＝0．21）。结论 sCTLA-4参与MG的发病机制,激素治疗、胸腺切除治疗减少sCT-LA-4表达。     

重症肌无力的分子生物学研究

重症肌无力的分子生物学研究许贤豪（北京协和医院神经科，北京１００７３０）分子生物学已深入到临床各学科，当前有种趋势，那就是用分子生物学观点来认识、用分子生物学手段来诊、疗临床各科有关疾病。现以重症肌无力为例来说明此问题。重症肌无力是一种累及神经—肌肉...     

兰州地区IL-15、IL-17A和IL-18基因多态性与复发性流产的相关性研究北大核心CSCD

目的:探讨IL-15、IL-17A和IL-18基因多态性与复发性流产(RSA)的关系。方法:SNaPshot法分析150例RSA患者(RSA组)和150例健康志愿者(对照组)中IL-15基因rs3806798、IL-17A基因rs2275913、IL-18基因rs1946518和rs187238位点多态性分布情况。结果:RSA组和对照组中4个单核苷酸多态性(SNPs)基因型频率和等位基因频率分布在两组间差异无统计学意义(P>0.05);隐性和显性遗传模式下对IL-15基因rs3806798、IL-17A基因rs2275913、IL-18基因rs1946518进行非条件Logistic回归分析,发现两组间差异均无统计学意义(P>0.05)。但IL-18基因的rs187238 SNP位点GG型、CG型、CC型3种基因型构成比在两组间差异有统计学意义(χ2=9.256,P=0.010)。显性、隐性遗传模式Logistic回归分析结果显示G等位型(GC+GG基因型)与RSA患病风险降低相关(χ2=4.303,OR=0.53,95%CI=0.29~0.97,P=0.038),但G等位基因不是RSA的保护性等位基因(χ2=2.275,OR=0.66,95%CI=0.38~1.14,P=0.132)。经连锁不平衡和单体型分析,rs1946518和rs187238存在完全连锁不平衡。TGTG单体型是RSA的保护因素,其OR(95%CI)=0.361(0.158~0.827)。结论:IL-18基因rs187238基因多态性与RSA患病风险相关。     

重症肌无力患者CD4、CD8细胞表面Fas分子的变化

为探讨Fas分子、CD4、CD8分子在重症肌无力发生中的作用 ,从胸腺及外周血分离单个核细胞 ,用荧光标记的单克隆抗体 (Fas FITC、CD4 PE、CD8 Cy )和流式细胞仪检测细胞表面Fas、CD4、CD8表达情况。结果发现 ,(1)MG患者外周血单个核细胞 (PBMC )CD4 + CD8 细胞明显低于对照组 (P <0 0 2 ) ;(2 )MG患者Fas分子在PBMC的表达率明显低于对照组 (P<0 0 2 ) ,Fas+ CD4 + 细胞也低于对照组 (P <0 0 5 ) ;(3)在Fas+ 的胸腺细胞中 ,MG患者的双阳性细胞显著低于对照组 (P <0 0 1) ,而CD4 CD8+ 细胞则高于对照组 (P <0 0 5 )。说明重症肌无力患者淋巴细胞存在Fas表达异常 ,表现为外周CD4 + 细胞活化后的凋亡障碍和胸腺CD4 + CD8+ 细胞凋亡障碍 ,可能与重症肌无力患者自身反应性T细胞的形成及疾病的发生有关     

重症肌无力患者骨髓、胸腺及外周血中IFN-γ、IL-4分泌细胞的检测

<正>重症肌无力（MG）是神经肌肉传递障碍的自身免疫性疾病,它是以抗乙酰胆碱受体（AchR）抗体为其主要的致病性抗体。目前认为该抗体的产生依赖于T淋巴细胞,T细胞所分泌的细胞因子参与了实验性自身免疫性重症肌无力（EAMG）的诱导和发病过程。干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）分别由Th1和 Th2亚型细胞所分泌,在机体免疫调节中起重要作用。本实验检测了早期发病的MG手术病人的骨髓、胸腺及外周血中IFN-γ和IL-4分泌细胞数,以期了解是否Th1及Th2均参与了MG的发病过程。     

HLA-DPB1等位基因与中国人重症肌无力相关性研究

目的：探讨在中国汉族人群中重症肌无力（ＭＧ）与ＨＬＡⅡ类基因的关联性。方法：随机抽取了５９例重症肌无力患者的外周血ＤＮＡ,用聚合酶链式反应－限制性片段长度多态性（ＰＣＲ－ＲＦＬＰ）方法,分析了其ＨＬＡ－ＤＰＢ１座位的基因型,并与正常对照进行比较。结果：发现ＨＬＡ－ＤＰＢ１＊０４０２与ＭＧ呈负相关（１１．８６％ｖｓ．２６．９２％;ｒｒ＝０．１８４３,Ｐｃ＝０．０２８９,ＰＦ＝０．２２０３）;＊１９０     

重症肌无力的认知功能研究

目的：研究重症肌无力（MG）的认知功能，方法：采用听觉词语记忆测验、数字符号转换、连线测验、逻辑记忆测验、Stroop色词测验（A，B，C）、言语流畅性、Boston命名测验等对36例MG全身划病例及33例眼肌型病例和36例健康对照组进行检查。结果：病例组与对照组比较，多项认知成绩下降，结论：MG存存认知功能障碍，提示MG可能存在CNS受累。     

胸腺扩大切除治疗重症肌无力围术期血清IL-17、IL-18、IL-27水平分析

目的探讨胸腺异常合并重症肌无力（MG）患者行胸腺扩大切除术后外周血血清IL—17、IL-18、IL-27等细胞因子的变化情况及其意义。方法选取16例未经免疫抑制治疗的MG患者行胸腔镜下胸腺扩大切除术,采用双抗体夹心ELISA法检测术前及术后第7El外周血血清IL-17、IL．18、IL．27水平,以15例健康献血者的血清IL-17、IL-18、IL-27水平作为正常对照。结果胸腺切除术后MG患者外周血IL-17、IL-18、IL-27水平普遍降低,外周血IL-17、IL-18、IL-27水平在胸腺瘤和胸腺增生两种病理类型中无明显差异。结论手术切除胸腺可降低MG患者外周血IL-17、IL-18、IL-27水平。     

重症肌无力患者记忆的初步研究

重症肌无力患者记忆的初步研究王文敏弓少霞王荪王毅昆明医学院第一附属医院神经内科目前认为重症肌无力（ｍｙａｓｔｈｅｎｉａｇｒａｖｉｓ，ＭＳ）是一种因体内产生的抗乙酰胆碱受体（ａｎｔｉ－ＡＣｈ－Ｒ）抗体作用于神经肌肉接头的运动终板，从而导致神经肌肉接头处...     

重症肌无力实验模型中胸腺细胞凋亡及其机制的研究

为观察重症肌无力中胸腺细胞凋亡的变化 ,制备了实验性自身免疫性重症肌无力 (EAMG )模型 ,采用切口及末端标记 (TUNEL )的方法观察了病程的不同阶段中胸腺细胞的凋亡情况。结果表明 ,EAMG的胸腺出现了细胞凋亡指数的减少 ,在免疫后第 4周 ,细胞凋亡指数减少最明显 (P <0 0 1)。进一步观察 ,可见实验鼠从第 4周开始 ,EAMG胸腺中Bcl 2的表达增加 ,而且细胞凋亡指数的减少与Bcl 2表达分数的增加呈明显的负相关。由此可见Bcl 2表达的增加 ,可导致EAMG胸腺中激活免疫细胞的正常凋亡过程抑制 ,可能是MG自身免疫发病机制之一。     

MuSK-mCherry融合荧光蛋白的构建及对重症肌无力患者MuSK抗体的检测

目的：构建肌肉特异性激酶（MuSK）与红色荧光蛋白（mCherry）的重组融合蛋白（MuSK-mCherry）,并作为抗原用于重症肌无力（ MG）患者血清中的MuSK抗体（ MuSKAb）的检测。方法：应用PCR技术,从含有mCherry 基因的载体pRSET-B扩增mCherry基因,经T载体（pGEM-T Easy Vector）克隆至含有MuSK细胞外区第22-452位氨基酸肽段基因的载体pMT/BiP/V5-His（ MuSK）上,构建MuSK-mCherry融合荧光蛋白基因。将重组载体转染果蝇S2细胞,表达产物以共聚焦显微镜检查。以MuSK-mCherry融合蛋白作为抗原,应用荧光免疫沉淀试验检测MG患者MuSKAb。结果：成功地构建了MuSK-mCherry融合蛋白基因,并得到了表达。经对已知MuSKAb阳性的MG患者血清中的MuSKAb的检测证实,构建的MuSK-mCherry融合蛋白在荧光免疫沉淀试验中可以检测到MuSKAb。结论：以MuSK细胞外肽段与mCherry构建的融合荧光蛋白作为抗原,可以用于MG患者血清中的MuSKAb的检测。     

The auto-antigen repertoire in myasthenia gravis

Myasthenia Gravis (MG) is an antibody-mediated autoimmune disorder affecting the postsynaptic membrane of the neuromuscular junction (NMJ). MG is characterized by an impaired signal transmission between the motor neuron and the skeletal muscle cell, caused by auto-antibodies directed against NMJ proteins. The auto-antibodies target the nicotinic acetylcholine receptor (nAChR) in about 90% of MG patients. In approximately 5% of MG patients, the muscle specific kinase (MuSK) is the auto-antigen. In the remaining 5% of MG patients, however, antibodies against the nAChR or MuSK are not detectable (idiopathic MG, iMG). Although only the anti-nAChR and anti-MuSK auto-antibodies have been demonstrated to be pathogenic, several other antibodies recognizing self-antigens can also be found in MG patients. Various auto-antibodies associated with thymic abnormalities have been reported, as well as many non-MG-specific auto-antibodies. However, their contribution to the cause, pathology and severity of the disease is still poorly understood. Here, we comprehensively review the reported auto-antibodies in MG patients and discuss their role in the pathology of this autoimmune disease.     

Diagnostic and clinical classification of autoimmune myasthenia gravis

Myasthenia gravis is characterized by muscle weakness and abnormal fatigability. It is an autoimmune disease caused by the presence of antibodies against components of the muscle membrane localized at the neuromuscular junction. In most cases, the autoantibodies are against the acetylcholine receptor (AChR). Recently, other targets have been described such as the MuSK protein (muscle-specific kinase) or the LRP4 (lipoprotein related protein 4). Myasthenia gravis can be classified according to the profile of the autoantibodies, the location of the affected muscles (ocular versus generalized), the age of onset of symptoms and thymic abnormalities.The disease generally begins with ocular symptoms (ptosis and/or diplopia) and extends to other muscles in 80% of cases. Other features that characterize MG include the following: variability, effort induced worsening, successive periods of exacerbation during the course of the disease, severity dependent on respiratory and swallowing impairment (if rapid worsening occurs, a myasthenic crisis is suspected), and an association with thymoma in 20% of patients and with other autoimmune diseases such as hyperthyroidism and Hashimoto's disease. The diagnosis is based on the clinical features, the benefit of the cholinesterase inhibitors, the detection of specific autoantibodies (anti-AChR, anti-MuSK or anti-LRP4), and significant decrement evidenced by electrophysiological tests.In this review, we briefly describe the history and epidemiology of the disease and the diagnostic and clinical classification. The neonatal form of myasthenia is explained, and finally we discuss the main difficulties of diagnosis.     

微小RNA与重症肌无力研究进展

微小RNA(miRNA)是一类进化上保守的内源性单链非编码小RNA分子,长度约为19 ～24个核苷酸,通过靶基因特异性结合从而导致mRNA降解或翻译抑制,在转录后水平调控基因表达.大量研究表明miRNA与多种人类自身免疫性疾病密切相关.重症肌无力(MG)是神经-肌肉接头处传递功能障碍的自身免疫性疾病.探究miRNA在MG发生发展中的分子机制有助于阐明MG的致病机制,为MG的分子诊断和个性化治疗提供重要依据.本文主要对miRNA与MG相关性的研究进展进行综述.     

胸腺切除治疗重症肌无力近期疗效及危象发生的影响因素

目的 分析胸腺切除治疗重症肌无力(MG)近期疗效及危象发生的影响因素.方法 回顾性分析436例胸腺切除治疗重症肌无力患者,对年龄、性别、病程、临床分型、胸腺病理类型、胸腺瘤病理分期、胸腺瘤病理类型等因素进行统计分析.结果 Ⅰ型患者术后近期症状稳定的比例较高[62.63%(186/297)],而ⅡA型以上患者术后近期改善及恶化的比例均较高,其差异有统计学意义(均为P＜0.05);各性别、年龄、病程、病理类型、胸腺瘤分期、胸腺瘤病理类型亚组间近期疗效差异无统计学意义(均为P＞0.05).Logistic回归分析显示,有无胸腺瘤(回归系数0.251)及肌无力分型(回归系数1.300)两个因素与术后危象发生有关(均为P＜0.05);伴有胸腺瘤的患者、ⅡA型以上患者术后发生危象概率明显偏高.结论 临床分型在ⅡA型以上MG患者术后近期症状易出现波动;患者性别、年龄、病程、病理类型、胸腺瘤分期、胸腺瘤病理类型与近期疗效无关;伴有胸腺瘤的MG患者、ⅡA型以上患者是术后危象发生的高危人群,术前要充分估计风险,围术期注意预防.     

Interleukin-4 receptor alpha polymorphisms in autoimmune myasthenia gravis in a Caucasian population

Autoimmune myasthenia gravis is a T-cell-dependent, antibody-mediated, rare neuromuscular disorder. Interleukin-4, acting via interleukin-4 receptor alpha, plays a pivotal role in B-cell differentiation and antibody production and has been implicated to influence disease progression in experimental autoimmune myasthenia gravis. Polymorphisms of the interleukin-4 receptor alpha gene have been shown to be associated with various autoimmune diseases. We compared the distribution of three polymorphisms of the interleukin-4 receptor alpha gene (S503P, rs1805015, Q576R, rs1801275, I75V, rs1805010), all affecting interleukin-4 signaling, in two cohorts of myasthenia gravis patients with ethnically matched controls. Although the distribution of the S503P and Q576R polymorphisms did not differ significantly between the groups, the frequency of the GG rare homozygote genotype of the I75V polymorphism was significantly higher in patients with myasthenia gravis. Our data suggest that the reduced responsiveness to interleukin-4 because the I75V polymorphism may contribute to the pathogenesis of myasthenia gravis.     

Complement and cytokine based therapeutic strategies in myasthenia gravis

Myasthenia gravis (MG) is a T cell–dependent and antibody-mediated disease in which the target antigen is the skeletal muscle acetylcholine receptor (AChR). In the last few decades, several immunological factors involved in MG pathogenesis have been discovered mostly by studies utilizing the experimental autoimmune myasthenia gravis (EAMG) model. Nevertheless, MG patients are still treated with non-specific global immunosuppression that is associated with severe chronic side effects. Due to the high heterogeneity of AChR epitopes and antibody responses involved in MG pathogenesis, the specific treatment of MG symptoms have to be achieved by inhibiting the complement factors and cytokines involved in anti-AChR immunity. EAMG studies have clearly shown that inhibition of the classical and common complement pathways effectively and specifically diminish the neuromuscular junction destruction induced by anti-AChR antibodies. The inborn or acquired deficiencies of IL-6, TNF-α and TNF receptor functions are associated with the lowest EAMG incidences. Th17-type immunity has recently emerged as an important contributor of EAMG pathogenesis. Overall, these results suggest that inhibition of the complement cascade and the cytokine networks alone or in combination might aid in development of future treatment models that would reduce MG symptoms with highest efficacy and lowest side effect profile.     

自身抗体与重症肌无力的研究新进展

重症肌无力（MG）主要是由乙酰胆碱受体抗体（AChRAb）介导的细胞免疫依赖补体参与的,针对神经肌肉接头突触后膜上乙酰胆碱受体的自身免疫性疾病。近年来对MG的发病机制作了大量的研究,发现自身抗体在MG的发病中有重要作用,如AChR抗体、阿理阿若碱（RyR）抗体和肌肉特异性酪氨酸激酶（MuSK）抗体等。由于MG病人血清中所含抗体的不同其临床特点不同,在发病机制中的作用也有差异,因而治疗MG前分析抗体水平可以提供更合理的治疗方式,通过抗体水平随访治疗效果。     

Serum cytokine and chemokine profiles in patients with myasthenia gravis

Myasthenia gravis (MG) is an autoimmune‐mediated inflammatory disease of the neuromuscular junction. Previous studies of animal MG models have suggested important roles of cytokines in MG pathogenesis, but adequate studies on cytokines in human MG are lacking. Using a multiplex suspension array system, we measured the serum levels of 27 cytokines/chemokines in 47 anti‐acetylcholine receptor antibody‐positive patients with MG and 20 normal controls (NC) to investigate the contribution of cytokines/chemokines toward MG pathogenesis. Correlations between clinical parameters and cytokine/chemokine levels in patients with MG were also examined. The serum levels of interleukin (IL)‐15 (mean ± standard deviation: 6·85 ± 6·97 pg/ml) and vascular endothelial growth factor (VEGF) (96·21 ± 71·60 pg/ml) significantly increased, whereas IL‐4 levels (3·57 ± 0·86 pg/ml) decreased in patients with MG compared with NC (IL‐15: 4·42 ± 1·55 pg/ml; VEGF: 63·51 ± 32·95 pg/ml; IL‐4: 4·15 ± 0·81 pg/ml, P < 0·05). In addition, eight cytokines (IL‐4, IL‐8, IL‐15, eotaxin, macrophage inflammatory protein‐1α, macrophage inflammatory protein‐1β, VEGF and IL‐1b) were significantly changed among MG patients with thymoma, MG patients without thymoma and NC (P < 0·05). Some cytokines, such as IL‐4, IL‐15, and VEGF, may play roles in the pathogenesis of MG.