A 型肉毒毒素治疗半侧面肌痉挛的临床疗效评价

目的：观察A型肉毒毒素对面肌痉挛患者的痉挛程度、抑郁症状和焦虑症状的改善。方法：对58例面肌痉挛患者进行局部注射A型肉毒毒素。在治疗前后对痉挛程度改善情况进行评定以及用汉密顿焦虑量表（HAMA）、汉密顿抑郁量表（HAMD）对焦虑状态、抑郁状态进行评分,并对药物的副作用进行观察。结果：A型肉毒毒素明显改善面肌痉挛患者的痉挛程度,治疗后2周的HAMA、HA肋评分较治疗前明显下降,且差异具有统计学意义（P〈0．01）。结论：局部注射A型肉毒毒素可迅速缓解或消除面肌痉挛患者肌肉痉挛及相关的抑郁和焦虑状态,提高患者的生活质量。     

肉毒毒素作用机制的研究进展

肉毒毒素(botulinum toxin,BTX)是肉毒梭状芽胞杆菌在生长繁殖过程中产生的一种外毒素,其通过抑制神经递质的释放而引起肌肉松弛型麻痹。在世界范围内,肉毒中毒的案例时有发生,病情严重的患者最终因呼吸衰竭而死亡。肉毒毒素相关产品在临床痉挛性疾病、腺体分泌过度、神经性疼痛的治疗及美容除皱等领域展现出广阔的应用前景。因而,肉毒毒素作用机制的研究在肉毒中毒的治疗以及临床新适应症的开发等方面具有重要意义。就肉毒毒素跨越小肠上皮细胞屏障的吸收及神经毒性作用机制的研究现状作一概述。     

肉毒毒素研究进展

肉毒毒素是肉毒梭菌产生的一种神经毒素,能够通过抑制神经肌肉接头处的乙酰胆碱释放而引起肌肉麻痹.肉毒毒素在培养液中以复合物形式存在,其中的毒性组分由3个非同源性结构域组成,是一种新型的金属蛋白酶.不同血清型的肉毒毒素能够特异性地作用于不同底物,这些底物在神经细胞的胞外分泌过程中发挥重要作用.肉毒毒素在胞吞胞吐机制的研究以及临床医学应用方面具有宝贵的价值.     

被肉毒杆菌毒素麻痹的肌肉随着神經再生的恢复

实驗在豚鼠进行,結果揭示,夹毁神经后运动神经末梢的再生在肉毒杆菌毒素中毒的肌肉与在正常肌肉完全一样地有效。用夹毁神经的办法,可以比较迅速地解除肉毒中毒肌肉的麻痹。本文结果,亦对肉毒的作用地点问题提供了新的佐証。     

婴儿肉毒中毒

<正> 一、肉毒中毒一般概念[1-3]肉毒毒素是由肉毒梭菌产生的一种高分子蛋白的神经毒素,它能引起死亡率很高的人和易感动物的肉毒中毒,尽管根据抗原的不同已将肉毒毒素分为A、B、C_1、C_2、D、E、F和G8个型,但它们都有抑制周围胆硷能运动神经末梢释放乙酰胆硷,妨碍神经传导,引起肌肉松弛性麻痹的药理作用。它们的结构和分子量也是相似的。     

肉毒毒素结构及其重组疫苗的相关研究进展

肉毒毒素是肉毒梭状杆菌产生的外毒素,有7种血清型(A～G).肉毒毒素属神经强毒,是目前已知的毒性最强的细菌蛋白质.作为重要的生物战剂之一,对肉毒毒素的研究已经相当深入,基本明确了各型肉毒毒素的基因序列、同源性和三维结构及毒素作用的本质和机理.随着国际恐怖活动的日益猖獗,针对肉毒毒素的检测和预防也备受关注,对其疫苗的探索已成为研究的焦点.本扼要介绍了肉毒毒素的结构、作用机制及其疫苗的相关研究进展.     

肉毒毒素和其它微生物神经毒素的性质及其在医学上的应用

引言 1989年12月,美国FDA批准A型肉毒毒素作为新药投产,以治疗12岁以上人的肌肉紊乱性斜视、偏侧面肌痉挛和眼睑痉挛。还可用于许多其它肌张力障碍和运动失调等疾病的实验性治疗。此举为毒素应用于人的神经和肌肉组织开辟了一个新领域。     

经肉毒杆菌毒素阻遏的接头在4-氨基吡啶、盐酸胍作用下神经末梢传导功能的恢复

A型肉毒杆菌毒素中毒大白鼠的膈神经膈肌标本,随着毒素作用的发展,细胞外记录的神经末梢动作电位与终板电位的平均振幅逐渐下降,直至完全消失,神经冲动不再能到达神经末梢。4-氨基吡啶、盐酸胍可恢复运动神经末梢传导冲动的能力,增加终板电位的量子含量,解除肉毒素产生的传递阻遏,恢复肌肉对间接刺激的收缩反应,而二性霉素对末梢电活动已消失的接头则无恢复作用。在上述资料基础上,对内毒素的作用机制和4-氨基吡啶等的抗肉毒效应进行了讨论。     

可美容的剧毒物--肉毒毒素

目前 ,一种被誉为“毒死皱纹”的毒针美容法正在风靡全球。这种奇妙的毒针是何物呢 ?是 A型肉毒毒素(botulinumtoxin,英文缩写为 BOTOX)。A型肉毒毒素 ,是世界上最毒的物质之一 ,一次注射过量足以致人死命 ;但如果将 0 .1m L 的肉毒毒素注射进皮肤 ,这个部位的皱纹竟会消失 6个月。A型肉毒毒素为什么能够消除皱纹呢 ?要解释这个问题 ,须要先了解皱纹形成的生理原因 :人体骨骼肌的运动依赖于运动神经的兴奋传递 ,这种兴奋传递来自于运动神经末梢释放的化学物质——乙酰胆碱的刺激 ,乙酰胆碱通过突触结构可刺激肌肉并使其收缩 ,从而产生了…     

凯式定氮法替代苦味酸法测定注射用A型肉毒毒素中明胶含量的可行性研究

目的 探究采用凯式定氮法替代苦味酸法测定注射用A型肉毒毒素(botulinum toxin type A for injection)明胶含量的可行性,以期对注射用A型肉毒毒素明胶含量测定方法的变更起到借鉴作用。方法 通过专属性、准确度、重复性、中间精密度和耐用性验证,确认采用凯式定氮法测定注射用A型肉毒毒素明胶含量的数据可靠性;对凯式定氮法与现有苦味酸法测定注射用A型肉毒毒素明胶含量的结果采用配对t检验和Bland-Altman分析进行统计学分析,确定2种测定方法的一致性。结果 注射用A型肉毒毒素中右旋糖酐20、蔗糖、A型肉毒毒素复合物对明胶含量测定无干扰,该方法专属性良好;准确度回收率位于95%～99%之间;重复性RSD为0.73%;中间精密度RSD为2.10%;且耐用性良好,可采用凯式定氮法进行注射用A型肉毒毒素明胶含量的测定。同时与现有苦味酸法相比,配对t检验无统计学差异;Bland-Altman分析,2种方法测量的差值均位于测量差均值的95%置信区间内,说明2种方法一致性良好。结论 可采用凯式定氮法替代苦味酸法进行注射用A型肉毒毒素明胶含量的测定。     

The relation between toxicity and toxin-related-antigen contents of Clostridium botulinum types C and D cultures as determined by mouse bioassay and ELISA

ELISA was tested for the adequacy to differentiate between botulinum type C1 and D toxins. The results presented in this paper indicate antigenic relationship between type C1 and D toxins. Furthermore, indications were obtained that the antigenicities of type C1 and of D toxins produced by different strains are not identical. Although ELISA can be used to differentiate type C1 and D toxins from those of other types, the assay has only limited value to differentiate between type C1 and D toxins. No parallel relationship between toxicity (mouse bioassay) and toxin-related-antigen contents (ELISA) of C. botulinum type C and D cultures was found.     

我国及部分国家B型肉毒毒素的中毒情况

肉毒毒素是自然界中已知毒性最强的毒素,通常被分为A～G共7个血清型,其中A、B、E型是最常见的人类中毒型别。肉毒中毒的流行特点与菌体的地域分布、各地居民的饮食习惯和社会活动都有一定关系。目前,除南极洲外的世界各大洲均有B型肉毒中毒的报道。我国、日本及欧洲B型肉毒中毒主要为家庭自制食物引发的食源性中毒,而美国则主要为婴儿肉毒中毒。近年来,创口型B型肉毒中毒与"注射型吸毒人员"的关联引起了研究者的注意。为了加深对B型肉毒中毒的了解,我们对我国及部分国家和地区的B型肉毒中毒情况做简要介绍。     

Learning from the past: historical aspects of bacterial toxins as pharmaceuticals

Botulinum neurotoxins are the most poisonous substances known to humankind, but also are the bacterial toxins most frequently used as pharmaceuticals to benefit humans. The discovery of botulinum toxins and development into a useful drug is unique and fascinating, dating back to the early 19th century, when Justinus Kerner first recognized that botulism was caused by a biological toxin and suggested its use for medicinal purposes. This was translated into reality in 1980, when Alan Scott for the first time used the toxins to successfully treat strabismus. Now a subset of botulinum toxins are widely used for cosmetic applications, treatment of various movement disorders, pain and many other syndromes, and further developments using other botulinum toxins or recombinant molecules engineered from subdomains are promising.     

Properties and use of botulinum toxin and other microbial neurotoxins in medicine.

Crystalline botulinum toxin type A was licensed in December 1989 by the Food and Drug Administration for treatment of certain spasmodic muscle disorders following 10 or more years of experimental treatment on human volunteers. Botulinum toxin exerts its action on a muscle indirectly by blocking the release of the neurotransmitter acetylcholine at the nerve ending, resulting in reduced muscle activity or paralysis. The injection of only nanogram quantities (1 ng = 30 mouse 50% lethal doses [U]) of the toxin into a spastic muscle is required to bring about the desired muscle control. The type A toxin produced in anaerobic culture and purified in crystalline form has a specific toxicity in mice of 3 x 10(7) U/mg. The crystalline toxin is a high-molecular-weight protein of 900,000 Mr and is composed of two molecules of neurotoxin (ca. 150,000 Mr) noncovalently bound to nontoxic proteins that play an important role in the stability of the toxic unit and its effective toxicity. Because the toxin is administered by injection directly into neuromuscular tissue, the methods of culturing and purification are vital. Its chemical, physical, and biological properties as applied to its use in medicine are described. Dilution and drying of the toxin for dispensing causes some detoxification, and the mouse assay is the only means of evaluation for human treatment. Other microbial neurotoxins may have uses in medicine; these include serotypes of botulinum toxins and tetanus toxin. Certain neurotoxins produced by dinoflagellates, including saxitoxin and tetrodotoxin, cause muscle paralysis through their effect on the action potential at the voltage-gated sodium channel. Saxitoxin used with anaesthetics lengthens the effect of the anaesthetic and may enhance the effectiveness of other medical drugs. Combining toxins with drugs could increase their effectiveness in treatment of human disease.     

Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse bioassay

A monoclonal antibody (BA11) has been produced against Clostridium botulinum type A neurotoxin by the fusion of myeloma cells (P3 NS1/1-Ag4-1) with spleen cells from BALB/c mice immunized with botulinum type A neurotoxoid. The antibody bound specifically to botulinum type A neurotoxin, showing no cross-reactivity with types B and E botulinum toxins or with any of several other bacterial toxins tested. The monoclonal antibody did not bind to botulinum type A neurotoxin which had been denatured with sodium dodecyl sulfate and bound only weakly to each of the separated heavy and light subunits of the neurotoxin, suggesting a conformational requirement for the antigenic determinant of the antibody. A sensitive immunoassay for C. botulinum type A toxin with monoclonal antibody BA11 in conjunction with an enzyme amplication system has been developed which allows detection of 5 to 10 mouse 50% lethal doses ml-1 of purified neurotoxin. The assay was equally sensitive when applied to the detection of crude toxin in food stuffs; the average value for the minimum level of detectable toxin in extracts of tinned salmon or corned beef was 9 +/- 3.1 mouse 50% lethal doses ml-1.     

Monoclonal antibody-based immunoassay for type A Clostridium botulinum toxin is comparable to the mouse bioassay.

A monoclonal antibody (BA11) has been produced against Clostridium botulinum type A neurotoxin by the fusion of myeloma cells (P3 NS1/1-Ag4-1) with spleen cells from BALB/c mice immunized with botulinum type A neurotoxoid. The antibody bound specifically to botulinum type A neurotoxin, showing no cross-reactivity with types B and E botulinum toxins or with any of several other bacterial toxins tested. The monoclonal antibody did not bind to botulinum type A neurotoxin which had been denatured with sodium dodecyl sulfate and bound only weakly to each of the separated heavy and light subunits of the neurotoxin, suggesting a conformational requirement for the antigenic determinant of the antibody. A sensitive immunoassay for C. botulinum type A toxin with monoclonal antibody BA11 in conjunction with an enzyme amplication system has been developed which allows detection of 5 to 10 mouse 50% lethal doses ml-1 of purified neurotoxin. The assay was equally sensitive when applied to the detection of crude toxin in food stuffs; the average value for the minimum level of detectable toxin in extracts of tinned salmon or corned beef was 9 +/- 3.1 mouse 50% lethal doses ml-1.     

Enzyme linked immunosorbent assay (ELISA) for detection of Clostridium botulinum type B toxin.

The enzyme-linked immunosorbent assay using different techniques has been applied to determine botulinum type B toxin. With the so-called "sandwich" technique, about 5,000 mouse ip LD50 of type B toxin can be detected. With the "double-sandwich" technique, about 400 mouse ip LD50 of toxin is detected and different commerical antisera are useful. For accurate quantification of botulinum toxins in culture filtrates, addition of EDTA to samples seems to be necessary. Cross-reactivity of the assay depends on the specificity of the antisera against botulinum type B toxin used and is almost eliminated with antiserum prepared against the toxic component of type B toxin.     

Development and optimization of immunoassays for the detection of botulinum toxins

Monoparametric immunoassay tests for detecting botulinum toxins types A and B and multiparametric assays for simultaneous detection of botulinum toxins type A and B have been developed. It is shown that the sensitivity of assays is affected by the size of nanoparticles of colloidal gold used as a marker of antibodies, load intensity of antibodies of colloidal gold in conjugates, the type of analytical membranes, as well as the chemical composition of buffer solutions used for the storage of conjugates and immunoassay analysis. The detection limit of monoparametric immunoassay tests is 0.5 ng/ml; that of multiparametric assays, 5.0 ng/ml. The developed immunoassay can be used for rapid assay of product quality, for grade control of botulinum toxins in pharmaceuticals, and environmental monitoring.     

Development and optimization of immunoassays for the detection of botulinum toxins

Monoparametric immunoassay tests for detecting botulinum toxins types A and B and multiparametric assays for simultaneous detection of botulinum toxins type A and B have been developed. It is shown that the sensitivity of assays is affected by the size of nanoparticles of colloidal gold used as a marker of antibodies, load intensity of antibodies of colloidal gold in conjugates, the type of analytical membranes, as well as the chemical composition of buffer solutions used for the storage of conjugates and immunoassay analysis. The detection limit of monoparametric immunoassay tests is 0.5 ng/ml; that of multiparametric assays, 5.0 ng/ml. The developed immunoassay can be used for rapid assay of product quality, for grade control of botulinum toxins in pharmaceuticals, and environmental monitoring.