肌力康饮对EAMG模型大鼠NMJ超微结构和外周细胞CD4~+细胞浓度的影响
摘要
目的:建立实验性自身免疫性重症肌无力(expermental autoimmune myasthenia gravis,EAMG)大鼠模型,通过观察肌力康饮对EAMG大鼠的临床症状、神经肌肉接头(Neuromusculai junction NMJ)超微结构和外周CD4~+细胞浓度的变化,探讨肌力康饮治疗重症肌无力的作用机制。
方法:健康雌性Lewis大鼠60只,随机分为空白组、模型组、阳性药物组、肌力康饮高剂量组、肌力康饮中剂量组、肌力康饮低剂量组,每组10只。除外空白组,其余各组大鼠用AchRα1 129-145肽段的抗原乳液造模。确认造模成功后,肌力康饮高、中、低剂量组分别以40g/(kg·d)、20g/(kg·d)、10g/(kg·d)的生药量灌胃,每日两次。阳性药物对照组给溴化吡啶斯的明4.05g/(kg·d),每日两次灌胃,空白组、模型组给蒸馏水灌胃,给药周期28天。给药结束后,对各组进行临床症状评分,并用电镜观察各组大鼠神经肌肉接头(NMJ)超微结构的变化、取肌肉组织用Motic -BA400显微镜于400倍视野下摄影,每组随机选取6个视野,用Motic image 3.2图像分析软件进行分析,计算其平均灰度值和平均光密度,反应CD4~+细胞浓度的变化。
结果:1、免疫动物6周后,进行模型制备的各组大鼠临床症状的评分、电镜观察大鼠神经肌肉接头超微结构与空白组比较有显著差异(P<0.01),以此判断造模成功,为研究MG的治疗提供了良好的实验动物模型。
2、治疗4周后,肌力康饮各组与模型组比较症状评分、促进神经肌肉接头(NMJ)新生轴突的产生,并增加突触小泡的数量,有统计学差异(P<0.05)、外周淋巴细胞CD4~+细胞减少(P<0.05),肌力康饮高剂量组与阳性药物组比较亦有统计学差异(P<0.05)。
结论:1、本实验用AchR-α1亚单位129-145片段免疫Lewis大鼠,诱导的EAMG大鼠模型在症状表现、神经肌肉接头超微结构方面的表现与人MG相似,初步判断模型成功,为研究肌力康饮的抗MG机制提供了条件。
2、肌力康饮治疗EAMG大鼠有效。分析其干预机制可能是通过可促进神经肌肉接头新生轴突的产生,并增加突触小泡的数量,外周淋巴细胞CD4~+细胞减少,从而达到治疗MG的作用。
Objective: To establish experimental autoimmune myasthenia gravis (expermental autoimmune myasthenia gravis, EAMG) rat model, by observing the health drink on the strength of clinical symptoms of EAMG rats, neuromuscular junction (Neuromusculai junction) ultrastructural and CD4~+ cells in peripheral blood lymphocytes change explore the strength Kang drink treatment of myasthenia gravis in rats.
Methods: 60 female Lewis rats were randomly divided into blank group, model group, positive medicine group, muscle strength and Sport drink high-dose group, muscle health drink in the dose group, muscle strength and Sport drink low-dose group, n = 10 only. Except the blank group, the remaining rats with AchRα1 129-145 peptide antigen latex modeling. After the success of recognition model, strength Kang drink high, medium and low-dose group, respectively 40g / (kg ? d), 20g / (kg ? d), 10g / (kg ? d) of the Health and dose gavage treatment, positive medicine the control group to pyridostigmine bromide, 4.05g / (kg ? d), twice a day, blank group, model group was given distilled water, treatment for 28 days.After administration of the clinical symptom score in each group and the rats in each group with the electron microscope neuromuscular junction (NMJ) ultrastructure changes. Muscle tissue obtained with the Motic-BA400 microscope under 400 times magnification in the photography, each group randomly selected six horizons, with the Motic image 3.2 image analysis software for analysis, calculation of the average gray value, the reaction changes in the concentration of CD4 ~+ cells.
Results: 1.The immune animals 6 weeks after the model prepared by rats in each group of clinical symptoms score, electron microscopy of rat neuromuscular junction ultrastructure compared with the control group were significantly different (P<0.01), in order to determine successful model for the study of the treatment of MG provides a good experimental animal model.
2.4 weeks after treatment, muscle strength and Sport drink each group compared with model group, symptom score, neuromuscular junction (NMJ) ultrastructure of a statistical difference (P < 0.05), In peripheral blood lymphocytes CD4~+ cells decreased (P<0.05),muscle strength and Sport drink high-dose group and the positive medicine group there are statistical differences (P<0.05).
Conclusion: 1.This experiment AchR-α1 subunit of 129-145 fragment immune Lewis rats, induced symptoms of EAMG rats in the performance of neuromuscular junction ultrastructure performance of the MG with people similar to the initial model to determine the success of To study the anti-drink muscle health conditions MG mechanism.
2. Muscle strength and effective health drink treatment of EAMG. Analysis of their mechanisms of intervention might be: through the neuromuscular junction can promote the production of new axons and increase the number of synaptic vesicles and Reduction of CD4~+ cells in peripheral blood lymphocytes to achieve the treatment of MG role.
方法:健康雌性Lewis大鼠60只,随机分为空白组、模型组、阳性药物组、肌力康饮高剂量组、肌力康饮中剂量组、肌力康饮低剂量组,每组10只。除外空白组,其余各组大鼠用AchRα1 129-145肽段的抗原乳液造模。确认造模成功后,肌力康饮高、中、低剂量组分别以40g/(kg·d)、20g/(kg·d)、10g/(kg·d)的生药量灌胃,每日两次。阳性药物对照组给溴化吡啶斯的明4.05g/(kg·d),每日两次灌胃,空白组、模型组给蒸馏水灌胃,给药周期28天。给药结束后,对各组进行临床症状评分,并用电镜观察各组大鼠神经肌肉接头(NMJ)超微结构的变化、取肌肉组织用Motic -BA400显微镜于400倍视野下摄影,每组随机选取6个视野,用Motic image 3.2图像分析软件进行分析,计算其平均灰度值和平均光密度,反应CD4~+细胞浓度的变化。
结果:1、免疫动物6周后,进行模型制备的各组大鼠临床症状的评分、电镜观察大鼠神经肌肉接头超微结构与空白组比较有显著差异(P<0.01),以此判断造模成功,为研究MG的治疗提供了良好的实验动物模型。
2、治疗4周后,肌力康饮各组与模型组比较症状评分、促进神经肌肉接头(NMJ)新生轴突的产生,并增加突触小泡的数量,有统计学差异(P<0.05)、外周淋巴细胞CD4~+细胞减少(P<0.05),肌力康饮高剂量组与阳性药物组比较亦有统计学差异(P<0.05)。
结论:1、本实验用AchR-α1亚单位129-145片段免疫Lewis大鼠,诱导的EAMG大鼠模型在症状表现、神经肌肉接头超微结构方面的表现与人MG相似,初步判断模型成功,为研究肌力康饮的抗MG机制提供了条件。
2、肌力康饮治疗EAMG大鼠有效。分析其干预机制可能是通过可促进神经肌肉接头新生轴突的产生,并增加突触小泡的数量,外周淋巴细胞CD4~+细胞减少,从而达到治疗MG的作用。
Objective: To establish experimental autoimmune myasthenia gravis (expermental autoimmune myasthenia gravis, EAMG) rat model, by observing the health drink on the strength of clinical symptoms of EAMG rats, neuromuscular junction (Neuromusculai junction) ultrastructural and CD4~+ cells in peripheral blood lymphocytes change explore the strength Kang drink treatment of myasthenia gravis in rats.
Methods: 60 female Lewis rats were randomly divided into blank group, model group, positive medicine group, muscle strength and Sport drink high-dose group, muscle health drink in the dose group, muscle strength and Sport drink low-dose group, n = 10 only. Except the blank group, the remaining rats with AchRα1 129-145 peptide antigen latex modeling. After the success of recognition model, strength Kang drink high, medium and low-dose group, respectively 40g / (kg ? d), 20g / (kg ? d), 10g / (kg ? d) of the Health and dose gavage treatment, positive medicine the control group to pyridostigmine bromide, 4.05g / (kg ? d), twice a day, blank group, model group was given distilled water, treatment for 28 days.After administration of the clinical symptom score in each group and the rats in each group with the electron microscope neuromuscular junction (NMJ) ultrastructure changes. Muscle tissue obtained with the Motic-BA400 microscope under 400 times magnification in the photography, each group randomly selected six horizons, with the Motic image 3.2 image analysis software for analysis, calculation of the average gray value, the reaction changes in the concentration of CD4 ~+ cells.
Results: 1.The immune animals 6 weeks after the model prepared by rats in each group of clinical symptoms score, electron microscopy of rat neuromuscular junction ultrastructure compared with the control group were significantly different (P<0.01), in order to determine successful model for the study of the treatment of MG provides a good experimental animal model.
2.4 weeks after treatment, muscle strength and Sport drink each group compared with model group, symptom score, neuromuscular junction (NMJ) ultrastructure of a statistical difference (P < 0.05), In peripheral blood lymphocytes CD4~+ cells decreased (P<0.05),muscle strength and Sport drink high-dose group and the positive medicine group there are statistical differences (P<0.05).
Conclusion: 1.This experiment AchR-α1 subunit of 129-145 fragment immune Lewis rats, induced symptoms of EAMG rats in the performance of neuromuscular junction ultrastructure performance of the MG with people similar to the initial model to determine the success of To study the anti-drink muscle health conditions MG mechanism.
2. Muscle strength and effective health drink treatment of EAMG. Analysis of their mechanisms of intervention might be: through the neuromuscular junction can promote the production of new axons and increase the number of synaptic vesicles and Reduction of CD4~+ cells in peripheral blood lymphocytes to achieve the treatment of MG role.
引文
[1]张海龙,赵晓刚等.中医辨证论治重症肌无力经验[J].中国医药指南,2008,1(1):208-209.
[2]金迪,张立德,刘宇.补脾益肾法治疗重症肌无力的机理探讨[J].辽宁中医药大学学报,2008,8(10):28-29.
[3]王殿华.陈金亮治疗重症肌无力经验[J].中医杂志,2006,8(47):583-593.
[4]李声岳.张燕平治疗眼肌型重症肌无力经验[J].中医杂志,2006,2(47):97.
[5]刘维,李相中.资生振痿丸治疗重症肌无力疗效观察[J].黑龙江中医药,2000,(5):30-31.
[6]张静生,孙巍,刘萍等.黄芪复方对实验性自身免疫性重症肌无力模型大鼠神经肌肉接头超微结构的影响[J].中国临床康复,2006,35(10):20-21.
[7]魏扬震,魏九康.针灸治疗重症肌无力26例[J].中国针灸,1999,19(6):345-346.
[8]邓光中.邓铁涛教授临证中脾胃学说的运用[J].新中医,2000,2(32):13-14.
[9]许凤全,陈金亮,黄涛.从奇经和络病理论治疗眼肌型重症肌无力的临床研究[J].新中医,2006,1(38):33-34.
[10]王永文,马泽洪.针药并用治疗眼型重症肌无力[J].针灸临床杂志,2004,20(9):15-16.
[11]李少芳,林卓鹏,林浩.针药结合治疗眼肌型重症肌无力100例疗效观察[J].新中医,2004,36(9):43-44.
[12]盛伟,李冬梅.重症肌无力的辨证治疗探析[J].实用中医内科杂志,2003,17(2):83.
[13]冯起国,崔红,林立泉,等.合谷刺法为主治疗眼肌型重症肌无力47例[J].中国针灸,1998,18(1):33-34.
[14]张临洪.重症肌无力危象的急诊处理[J].临床内科杂志,1991,8(6):27.
[15]游鼎元,徐得忠.59万人口重症肌无力流行病学调查报告[J].中国神经精神疾病杂志,1988,14(4):231.
[16]吴江,贾建平,崔丽英.神经病学[M].北京:人民卫生出版社,2005,341-346.
[17]王霞.阮丽荣.张清勇等.重症肌无力患儿胸腺细胞体外增殖和CD4、CD8、Fas的测定[J].郑州大学学报,2005,1(40):10-11.
[18]张星虎,许贤豪,殷剑,等.免疫学因素对重症肌无力患者外周血单个核细胞糖皮质激素受体的影响[J].中华微生物学和免疫学杂志,2002,5(22):559-560.
[19]国红,孙宏,许贤豪,等.重症肌无力特异性细胞免疫应答[J].免疫学杂志,2000,2(16):114-115.
[20]. Lennon V A,Lindstrom J M,Seybold M E.Experimental autoimmune myasthenia:a model of myasthenia gravis in rats and guinea pigs[J].Exp Med,1975;141:1365-1375.
[21]李婉宜,李明远,陈平,等.同种异型T细胞抑制全身型MG患者AchR-Ab产生的研究[J].上海免疫学杂志,2000,5(20):289-290.
[22]李作孝,赖成虹,荣本兵等.重症肌无力患者外周血T淋巴细胞亚群凋亡的研究[J].中华神经医学杂志,2006,8(5):803-804.
[23]徐东,扬海卫.重症肌无力的发病机制[J].山东医药,2004,22(44):59.
[24]解燕春,李承晏.重症肌无力治疗新进展[J].临床内科杂志, 2005,6(22):374-375.
[25] Richman DP,Agius MA.Treatmet of autoimmune myasthenia gravis neurology,2003,61: 1652-1661.
[26] Gronseth GS,Barohn RJ.Thynectomy for non-thymomatous autoimmune Myasthenia granis(an evidence-based review).Neurology,2000,55:7-15.
[27] Sanders DB. Clinical neurophysiology of disorders of the neuromuscular junction [J]. J Clin Neurophysiol, 1993,10(2):167-180.
[28] CuiLiying,Guan Yuzhou,WangHan,et a.l Single fiber electrom yography in the diagnosis of ocularm yasthenia gravis: reportof 90 cases [J]. Chinese Medical Journal,2004,117 (6): 848-851.
[29]许贤豪.重症肌无力.神经免疫学[M].北京医科大学、中国协和医科大学联合出版社,1992.113-157.
[30]郝志波,郭晨云,等.基因疫苗pcDNA-AChRα211免疫小鼠建立重症肌无力动物模型[J].中国免疫学杂志,2006,22:217.
[31]吴怀国,陈荣志,许文华,等.乙酰胆碱受体α亚基125-147肽段制作实验性重症肌无力动物模型的研究[J].临床神经病学杂志,2006,19(5):369.
[32]孙巍,张静生,刘萍,等.实验性自身免疫性重症肌无力( EAMG)大鼠模型的研究[J].中医药学刊,2005,5:812-814.
[33]彭秀玲,袁汉英,谢毅,等.基因工程实验技术[M].第2版.长沙;湖南科学技术出版社,1998,261-264.
[34] WANG Yun-fu,SUN Sheng-gang,HE Guo-hou,et al.Study on animal of experimental autoimmune myasthenia gravis in rats.China Journal of Moderm Medicine,2006,12:3682.
[35]战克斌,卜碧涛,李志军等.乙酰胆碱受体诱导实验性自身免疫性重症肌无力模型的建立[J].中国神经免疫学和神经病学杂志,2006,13:18-20.
[36]李秀华,程焱,杨丽等.被动转移法致自身免疫性重症肌无力兔模型的建立[J].中国神经精神疾病杂志,2009,1(23):12-13.
[37]高波廷,李保华,杨明山,等.重症肌无力被动转移模型的研究[J].Chin J Clin Neurusci 2001,9(2),130-132.
[38] Grans YMF,Venschuuren JJGM,Bos NA,et al.VH gene family utilization of anti-AChR antibodies in experimental autoimmune myasthenia gravis[J].J Neuroimmunol,1993,43(1-2):113-124 .
[39] Zhang Xu,Mario Losen,Marc De Baets.The Effect of Anti-acetylcholine Receptor Monoclonal Antibody A7 in Experimental Autoimmune Myasthenia Gravis.Chin[J] Neuroimmunol Neurol 2005,12(2):84.
[40]孟繁平,杨康娟.人免疫球蛋白转基因小鼠自身免疫性重症肌无力模型的建立[J].中华微生物学和免疫学杂志,2005,25(2):170.
[41] Christadoss P,Poussin M,Deng C.Animal models of myasthenia gravis[J].Clin Immunol,2000;94:76-87.
[42] Im SH,Barchan D,Souronjon MC,et al.Role of tolerogen conformation in induction of oral tolerance in experimental autoimmune myasthenia gravis,J Immunol,2000,165(7):3599-3605.
[43]唐冰杉,扬明山,曹学兵.实验性自身免疫性重症肌无力动物模型的研究概况[J].国外医学神经病学神经外科学分册,2002,29(3):267.
[44]崔丽英,刘明生.重症肌无力的电生理诊断[J].中国实用内科杂志,2006, 4( 26) :249-250.
[45]刘萍,张静生.黄芪复方对EAMG大鼠血清IL-4、IFN-γ、TGF-β水平的影响研究[J].中医药学刊,2004,22(9):1611.
[46]孙博,李呼论,赵吉成,等.实验性重症肌无力大鼠模型的建立及巨噬细胞在发病机制中的作用探讨[J].中国免疫学杂志,2006,22:1010-1013.
[47]王英鹏,李柱一.重症肌无力中枢损害脾脏改变及细胞调亡相关蛋白表达[J].第四军医大学学报,2003,24(15):1352-1355.
[48]徐冬波,戴静芝.强力方对实验性自身免疫性重症肌无力细胞因子含量的影响[J].上海中医药杂志,2000,10:40-43.
[49]刘炳凯,曹蓓蓓.曹鸣高治疗痿证的经验[J].四川中医,2000,2(18):1-2.
[50]解燕春,李承晏.重症肌无力治疗新进展[J].临床内科杂志,2005,6(22):374-375.
[51]丛志强,李文馨.重症肌无力的治疗进展[J].医师进修杂志,2000,8(23):11-12.
[52]莫雪安.重症肌无力的干细胞治疗现状[J].临床内科杂志,2005,6(22):371-372.60.王永忠,甄文俊,严伟忠等.
[53]310例重症肌无力的外科治疗[J].中华外科杂志,2003,8(41):607-608.
[54]乌立晖,徐志飞,赵学维等.胸腺瘤合并重症肌无力的外科治疗[J].实用肿瘤杂志,2004,3(19):246-247.
[55]陈志明,姚亚其,庞烈文等.Ⅲ型危重重症肌无力的外科治疗[J].上海医学,2000,1(23):14-15.
[56]刘雅彬,黄壮士,张玉斌,等.改良手术方法治疗重症肌无力38例[J].郑州大学学报,2004,4(39):706-707.
[57]张鹏,刘懿,鄢盛尧,等.重症肌无力外科治疗的临床分析[J].中华胸心血管外科杂志,2004,5(20):299.
[58]刘阳,张清,周乃康,等.重症肌无力的外科治疗[J].军医进修学报,2005,26(4):296-297.
[59]杨明山,曹学兵.重症肌无力研究现状与展望[J].中国神经免疫学和神经病学杂志,2002,9(2):63-64.
[60]王云甫,孙圣刚,曹学兵等.实验性重症肌无力大鼠模型的研究[J].中国神经免疫学和神经病学杂志,2003,4(10):242-243.
[61]杨春晓,王化冰,王维治.实验性自身免疫性重症肌无力模型的制备[J].中国临床康复,2006,28(22):213.
[62]申志强,孙俊秀,黄立新.黄芪治疗选择性IgA缺乏的疗效[J].新药与临床,1997,16(4):246-247.
[63]张太阳,张启发.黄芪对Graves病患者T淋巴细胞亚群的影响[J].江西医学院学报,1995,35(2):29.
[64]张艳,梁华平.黄芪多糖对烧伤小鼠细胞免疫功能的作用及机理探讨[J].中国实验临床免疫学杂志,1995,7(2):41.
[65]焦红军.党参的药理作用及其临床应用[J].临床医学,2005,4(25):92-93.
[66]张兆林.党参多糖成分及其免疫药理作用研究[J].兰州医学院学报,1988,11(3):14-5.
[67]苗明三.薏苡仁多糖对环磷酰胺致免疫抑制小鼠免疫功能的影响[J].中医药学报,2002,30(5):49.
[68] Inoue M,Fulli Y,Okmura M,et al.Mmture CD4 simge positive thymocyte in human thymoma:T cell may differentiate in the thmicepithelial cell tumor pathobiology.Neurology,1997,65:216-222.
[69]杨颖.重症肌无力发病机制的研究现状与展望.北京医学, 2004 ,26(5):349-350.
[70] Zhang GX,Xiao B G,Bakhiel M,et al.Both CD4+ and CD8+ T cells are essential to induce experimental autoummune myasthenia gravie[J].J Exp Med,1996; 184: 349.
[2]金迪,张立德,刘宇.补脾益肾法治疗重症肌无力的机理探讨[J].辽宁中医药大学学报,2008,8(10):28-29.
[3]王殿华.陈金亮治疗重症肌无力经验[J].中医杂志,2006,8(47):583-593.
[4]李声岳.张燕平治疗眼肌型重症肌无力经验[J].中医杂志,2006,2(47):97.
[5]刘维,李相中.资生振痿丸治疗重症肌无力疗效观察[J].黑龙江中医药,2000,(5):30-31.
[6]张静生,孙巍,刘萍等.黄芪复方对实验性自身免疫性重症肌无力模型大鼠神经肌肉接头超微结构的影响[J].中国临床康复,2006,35(10):20-21.
[7]魏扬震,魏九康.针灸治疗重症肌无力26例[J].中国针灸,1999,19(6):345-346.
[8]邓光中.邓铁涛教授临证中脾胃学说的运用[J].新中医,2000,2(32):13-14.
[9]许凤全,陈金亮,黄涛.从奇经和络病理论治疗眼肌型重症肌无力的临床研究[J].新中医,2006,1(38):33-34.
[10]王永文,马泽洪.针药并用治疗眼型重症肌无力[J].针灸临床杂志,2004,20(9):15-16.
[11]李少芳,林卓鹏,林浩.针药结合治疗眼肌型重症肌无力100例疗效观察[J].新中医,2004,36(9):43-44.
[12]盛伟,李冬梅.重症肌无力的辨证治疗探析[J].实用中医内科杂志,2003,17(2):83.
[13]冯起国,崔红,林立泉,等.合谷刺法为主治疗眼肌型重症肌无力47例[J].中国针灸,1998,18(1):33-34.
[14]张临洪.重症肌无力危象的急诊处理[J].临床内科杂志,1991,8(6):27.
[15]游鼎元,徐得忠.59万人口重症肌无力流行病学调查报告[J].中国神经精神疾病杂志,1988,14(4):231.
[16]吴江,贾建平,崔丽英.神经病学[M].北京:人民卫生出版社,2005,341-346.
[17]王霞.阮丽荣.张清勇等.重症肌无力患儿胸腺细胞体外增殖和CD4、CD8、Fas的测定[J].郑州大学学报,2005,1(40):10-11.
[18]张星虎,许贤豪,殷剑,等.免疫学因素对重症肌无力患者外周血单个核细胞糖皮质激素受体的影响[J].中华微生物学和免疫学杂志,2002,5(22):559-560.
[19]国红,孙宏,许贤豪,等.重症肌无力特异性细胞免疫应答[J].免疫学杂志,2000,2(16):114-115.
[20]. Lennon V A,Lindstrom J M,Seybold M E.Experimental autoimmune myasthenia:a model of myasthenia gravis in rats and guinea pigs[J].Exp Med,1975;141:1365-1375.
[21]李婉宜,李明远,陈平,等.同种异型T细胞抑制全身型MG患者AchR-Ab产生的研究[J].上海免疫学杂志,2000,5(20):289-290.
[22]李作孝,赖成虹,荣本兵等.重症肌无力患者外周血T淋巴细胞亚群凋亡的研究[J].中华神经医学杂志,2006,8(5):803-804.
[23]徐东,扬海卫.重症肌无力的发病机制[J].山东医药,2004,22(44):59.
[24]解燕春,李承晏.重症肌无力治疗新进展[J].临床内科杂志, 2005,6(22):374-375.
[25] Richman DP,Agius MA.Treatmet of autoimmune myasthenia gravis neurology,2003,61: 1652-1661.
[26] Gronseth GS,Barohn RJ.Thynectomy for non-thymomatous autoimmune Myasthenia granis(an evidence-based review).Neurology,2000,55:7-15.
[27] Sanders DB. Clinical neurophysiology of disorders of the neuromuscular junction [J]. J Clin Neurophysiol, 1993,10(2):167-180.
[28] CuiLiying,Guan Yuzhou,WangHan,et a.l Single fiber electrom yography in the diagnosis of ocularm yasthenia gravis: reportof 90 cases [J]. Chinese Medical Journal,2004,117 (6): 848-851.
[29]许贤豪.重症肌无力.神经免疫学[M].北京医科大学、中国协和医科大学联合出版社,1992.113-157.
[30]郝志波,郭晨云,等.基因疫苗pcDNA-AChRα211免疫小鼠建立重症肌无力动物模型[J].中国免疫学杂志,2006,22:217.
[31]吴怀国,陈荣志,许文华,等.乙酰胆碱受体α亚基125-147肽段制作实验性重症肌无力动物模型的研究[J].临床神经病学杂志,2006,19(5):369.
[32]孙巍,张静生,刘萍,等.实验性自身免疫性重症肌无力( EAMG)大鼠模型的研究[J].中医药学刊,2005,5:812-814.
[33]彭秀玲,袁汉英,谢毅,等.基因工程实验技术[M].第2版.长沙;湖南科学技术出版社,1998,261-264.
[34] WANG Yun-fu,SUN Sheng-gang,HE Guo-hou,et al.Study on animal of experimental autoimmune myasthenia gravis in rats.China Journal of Moderm Medicine,2006,12:3682.
[35]战克斌,卜碧涛,李志军等.乙酰胆碱受体诱导实验性自身免疫性重症肌无力模型的建立[J].中国神经免疫学和神经病学杂志,2006,13:18-20.
[36]李秀华,程焱,杨丽等.被动转移法致自身免疫性重症肌无力兔模型的建立[J].中国神经精神疾病杂志,2009,1(23):12-13.
[37]高波廷,李保华,杨明山,等.重症肌无力被动转移模型的研究[J].Chin J Clin Neurusci 2001,9(2),130-132.
[38] Grans YMF,Venschuuren JJGM,Bos NA,et al.VH gene family utilization of anti-AChR antibodies in experimental autoimmune myasthenia gravis[J].J Neuroimmunol,1993,43(1-2):113-124 .
[39] Zhang Xu,Mario Losen,Marc De Baets.The Effect of Anti-acetylcholine Receptor Monoclonal Antibody A7 in Experimental Autoimmune Myasthenia Gravis.Chin[J] Neuroimmunol Neurol 2005,12(2):84.
[40]孟繁平,杨康娟.人免疫球蛋白转基因小鼠自身免疫性重症肌无力模型的建立[J].中华微生物学和免疫学杂志,2005,25(2):170.
[41] Christadoss P,Poussin M,Deng C.Animal models of myasthenia gravis[J].Clin Immunol,2000;94:76-87.
[42] Im SH,Barchan D,Souronjon MC,et al.Role of tolerogen conformation in induction of oral tolerance in experimental autoimmune myasthenia gravis,J Immunol,2000,165(7):3599-3605.
[43]唐冰杉,扬明山,曹学兵.实验性自身免疫性重症肌无力动物模型的研究概况[J].国外医学神经病学神经外科学分册,2002,29(3):267.
[44]崔丽英,刘明生.重症肌无力的电生理诊断[J].中国实用内科杂志,2006, 4( 26) :249-250.
[45]刘萍,张静生.黄芪复方对EAMG大鼠血清IL-4、IFN-γ、TGF-β水平的影响研究[J].中医药学刊,2004,22(9):1611.
[46]孙博,李呼论,赵吉成,等.实验性重症肌无力大鼠模型的建立及巨噬细胞在发病机制中的作用探讨[J].中国免疫学杂志,2006,22:1010-1013.
[47]王英鹏,李柱一.重症肌无力中枢损害脾脏改变及细胞调亡相关蛋白表达[J].第四军医大学学报,2003,24(15):1352-1355.
[48]徐冬波,戴静芝.强力方对实验性自身免疫性重症肌无力细胞因子含量的影响[J].上海中医药杂志,2000,10:40-43.
[49]刘炳凯,曹蓓蓓.曹鸣高治疗痿证的经验[J].四川中医,2000,2(18):1-2.
[50]解燕春,李承晏.重症肌无力治疗新进展[J].临床内科杂志,2005,6(22):374-375.
[51]丛志强,李文馨.重症肌无力的治疗进展[J].医师进修杂志,2000,8(23):11-12.
[52]莫雪安.重症肌无力的干细胞治疗现状[J].临床内科杂志,2005,6(22):371-372.60.王永忠,甄文俊,严伟忠等.
[53]310例重症肌无力的外科治疗[J].中华外科杂志,2003,8(41):607-608.
[54]乌立晖,徐志飞,赵学维等.胸腺瘤合并重症肌无力的外科治疗[J].实用肿瘤杂志,2004,3(19):246-247.
[55]陈志明,姚亚其,庞烈文等.Ⅲ型危重重症肌无力的外科治疗[J].上海医学,2000,1(23):14-15.
[56]刘雅彬,黄壮士,张玉斌,等.改良手术方法治疗重症肌无力38例[J].郑州大学学报,2004,4(39):706-707.
[57]张鹏,刘懿,鄢盛尧,等.重症肌无力外科治疗的临床分析[J].中华胸心血管外科杂志,2004,5(20):299.
[58]刘阳,张清,周乃康,等.重症肌无力的外科治疗[J].军医进修学报,2005,26(4):296-297.
[59]杨明山,曹学兵.重症肌无力研究现状与展望[J].中国神经免疫学和神经病学杂志,2002,9(2):63-64.
[60]王云甫,孙圣刚,曹学兵等.实验性重症肌无力大鼠模型的研究[J].中国神经免疫学和神经病学杂志,2003,4(10):242-243.
[61]杨春晓,王化冰,王维治.实验性自身免疫性重症肌无力模型的制备[J].中国临床康复,2006,28(22):213.
[62]申志强,孙俊秀,黄立新.黄芪治疗选择性IgA缺乏的疗效[J].新药与临床,1997,16(4):246-247.
[63]张太阳,张启发.黄芪对Graves病患者T淋巴细胞亚群的影响[J].江西医学院学报,1995,35(2):29.
[64]张艳,梁华平.黄芪多糖对烧伤小鼠细胞免疫功能的作用及机理探讨[J].中国实验临床免疫学杂志,1995,7(2):41.
[65]焦红军.党参的药理作用及其临床应用[J].临床医学,2005,4(25):92-93.
[66]张兆林.党参多糖成分及其免疫药理作用研究[J].兰州医学院学报,1988,11(3):14-5.
[67]苗明三.薏苡仁多糖对环磷酰胺致免疫抑制小鼠免疫功能的影响[J].中医药学报,2002,30(5):49.
[68] Inoue M,Fulli Y,Okmura M,et al.Mmture CD4 simge positive thymocyte in human thymoma:T cell may differentiate in the thmicepithelial cell tumor pathobiology.Neurology,1997,65:216-222.
[69]杨颖.重症肌无力发病机制的研究现状与展望.北京医学, 2004 ,26(5):349-350.
[70] Zhang GX,Xiao B G,Bakhiel M,et al.Both CD4+ and CD8+ T cells are essential to induce experimental autoummune myasthenia gravie[J].J Exp Med,1996; 184: 349.