A型肉毒毒素在国内外的临床应用

在介绍Ａ型肉毒毒素结构、功能及性质的基础上,对其应用于临床治疗中的国内外情况进行了综述,旨在为此毒素的应用提供理论依据及实践经验,并指出以后应深入研究的重点,以使该毒素的临床应用更趋完美。     

C型肉毒毒素与C型肉毒中毒

本文介绍了Ｃ型肉毒毒素的纯化、生物学特性,并从流行病学角度对动物Ｃ型肉毒中毒的发生、中毒机理及临床表现、诊断、治疗及预防等进行了概述。对近年来Ｃ型肉毒毒素在分子水平、作用机理等方面的研究进展做了简要介绍。     

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.     

苦味酸法测定A型肉毒毒素中的明胶含量

目的注射用A型肉毒毒素中加入明胶作为稳定剂,建立并验证定量检测注射用A型肉毒毒素中明胶含量的方法。方法通过苦味酸与明胶特异性作用,产生强的吸收,采用紫外-可见分光光度计用外标法来检测注射用A型肉毒毒素中的明胶含量。结果通过一定浓度苦味酸与明胶作用,用分光光度计在520 nm处测定吸光度,检测限可以达到2.5 mg/L,检测范围可以达到10~100 mg/L。结论该方法具有良好的特异性、准确度、精密度和灵敏度,对于测定注射用A型肉毒毒素中的明胶含量有较好的参考价值。     

A型肉毒毒素轻链基因的克隆及其结构分析

以献报道的A型肉毒毒素基因全序列为标准,设计并合成一对引物,自肉毒梭菌基因组中扩增出肉毒毒素轻链基因片段,并将扩增产物与pGEM—T载体在体外连接,构建测序重组质粒,进行测序和基因结构分析。PCR扩增获得了产物为1 364bp的DNA片段,测序结果与DNA数据库对照检索分析证明,此基因片段与GenBank中的A型肉毒毒素LC基因的一致性达99．9％以上,可以认为克隆的基因为A型肉毒毒素LC基因。     

肉毒杆菌二元毒素和细胞外酶

对肉毒中毒和肉毒毒蛋白的研究已有近百年历史,肉毒杆菌神经毒素（ＢｏＮＴ）作用机制研究的进展已使它成分为分析神经递质释放的胞吐分子机制和用于临床治疗的有用工具。近年发现,肉毒梭状芽胞杆菌Ｃ和Ｄ型菌株除分泌ＢｏＮＴ外,尚合成另外两类与ＢｏＮＴ结构和生物活必完全不同的蛋白质,肉毒二元毒素和肉毒细胞外酶,它们分别影响细胞骨架和细胞的生长发育,也成为分析相关生命过程的有效工具药,本文综述了这两类蛋白质的有关     

治疗用A型肉毒毒素的制备及其质量控制

我国治疗用Ａ型肉毒毒素采用酸等电点沉淀,磷酸盐提取,核糖核酸酶处理,ＤＥＡＥ Ａ５０离子交换层析,硫酸铵浓缩及自然结晶等程序从Ａ型肉毒梭菌培养物中提取,并经稀释、冻干而成。它毒力强、纯度高、性能稳定,宜于长期保存。经生化、免疫学测定,毒素系神经毒素和血凝素的复合体,纯度为２５～３．０×１０７ＬＤ５０（小鼠,下同）／ｍｇｐｒ,ＯＤ２６０／ＯＤ２８０≤０．５５不仅达到了美国ＦＤＡ对注射用Ａ型肉毒毒素的质量要求,而且在冻干损失,稳定性方面还优于美国制品。     

A型肉毒毒素联合肌电生物反馈治疗脑卒中后上肢肌痉挛的临床疗效分析

目的:分析A型肉毒毒素与肌电生物反馈联合用于治疗脑卒中后上肢肌痉挛的临床疗效。方法:选择2014年10月至2016年12月辽宁本溪市中心医院和北京博爱医院收治的84例脑卒中后上肢肌痉挛患者,并将其随机分为观察组和对照组,每组42例。两组患者均首先接受常规康复训练,随后对照组加用肌电生物反馈治疗,观察组患者在对照组的基础之上注射A型肉毒毒素。采用改良Ashworth痉挛量表(MAS)比较两组治疗前后的上肢肌痉挛改善情况,采用Fugl-Meyer评定量表评价患者上肢运动功能,采用量角器测量治疗前后患者的腕关节主动活动范围,采用改良的Barthel指数评价患者治疗前后的日常生活能力。结果:治疗后,观察组的上肢痉挛改善总有效率显著高于对照组(P0.05);治疗2周和4周时,两组的Fugl-Meyer评分、腕关节主动活动范围、改良的Barthel指数(MBI)均较治疗前显著升高(P0.05),且观察组治疗后2周和4周的Fugl-Meyer评分、腕关节主动活动范围、改良的Barthel指数(MBI)均显著高于对照组(P0.05)。结论:A型肉毒毒素与肌电生物反馈联合用于治疗脑卒中后上肢肌痉挛的临床疗效显著,可有效降低患者上肢痉挛状态,改善上肢和腕部运动能力,提高患者的日常生活能力。     

IEF纯化重组A型肉毒毒素保护性抗原

在大肠杆菌中高效表达的重组A型肉毒毒素保护性抗原（rBoNTaH468),是以包涵体形式存在,将表达菌株发酵后,裂解菌体,制备包涵体,溶解后的包涵体溶液经样品处理,通过等地电聚焦制备型电泳纯化,纯化的重组A型肉毒毒素保护性抗原（rBoNTaH468)纯度高于90%,产量及回收率高,纯化的重组表达产物酶联检测具有结合活性,这为下一步A型肉毒毒素抗毒素的研制打下基础。     

Muscle architecture of the upper limb in the orangutan

We dissected the left upper limb of a female orangutan and systematically recorded muscle mass, fascicle length, and physiological cross-sectional area (PCSA), in order to quantitatively clarify the unique muscle architecture of the upper limb of the orangutan. Comparisons of the musculature of the dissected orangutan with corresponding published chimpanzee data demonstrated that in the orangutan, the elbow flexors, notably M. brachioradialis, tend to exhibit greater PCSAs. Moreover, the digital II-V flexors in the forearm, such as M. flexor digitorum superficialis and M. flexor digitorum profundus, tend to have smaller PCSA as a result of their relatively longer fascicles. Thus, in the orangutan, the elbow flexors demonstrate a higher potential for force production, whereas the forearm muscles allow a greater range of wrist joint mobility. The differences in the force-generating capacity in the upper limb muscles of the two species might reflect functional specialization of muscle architecture in the upper limb of the orangutan for living in arboreal environments.     

脑卒中痉挛性患者经颅磁刺激联合物理治疗后上肢功能恢复状况的临床观察

摘要 目的：探讨脑卒中痉挛性患者经颅磁刺激联合物理治疗后上肢功能改善状况的临床观察。方法：选取我院2018年1月到2020年1月共收治的80例脑卒中患者,所有患者均出现不同部位痉挛现象,将患者随机分为观察组与对照组,每组40例。给予对照组患者常规治疗与康复训练,观察组患者在常规治疗基础上应用低频重复经颅刺激联合肌电生物反馈模式下的康复训练。对比两组患者的治疗效果与上肢功能改善情况。结果：观察组治疗总有效率95.00 %,高于对照组72.50 %（P＜0.05）;治疗前两组患者的NIHSS评分应用神经功能缺损量表（Neurological deficit scale,NIHSS）、MMSE评分应用认知功能量表（Cognitive function scale,MMSE）、ADL评分应用日常生活能力量表（Activities of daily living scale,ADL）评分对比无显著差异（P＞0.05）,治疗后,观察组的NIHSS评分低于对照组,ADL评分高于对照组（P＜0.05）;治疗前两组患者的肱二头肌和肱三头肌均方根值（Root mean square,RMS）对比无显著差异（P＞0.05）,治疗后,观察组患者的肱二头肌RMS低于对照组,肱三头肌RMS高于对照组（P＜0.05）;治疗前两组患者的运动功能评估表中的上肢功能部分（Upper limb function in motor function assessment table,FMA-UE）、手部精细化动作及上肢功能测量表（Measurement table of hand fine movement and upper limb function,Carroll）评分对比无显著差异（P＞0.05）,治疗后,观察组患者的FMA-UE、Carroll评分高于对照组（P＜0.05）。结论：对脑卒中上肢痉挛患者在常规治疗与康复训练的基础上应用低频重复经颅刺激联合肌电生物反馈模式下的康复训练,虽然对患者的认知功能无明显影响,但是能提升患者上肢痉挛的治疗效果,促进患者上肢功能恢复,提高生活能力,值得临床应用推广。     

A型肉毒毒素注射在改善面上部动力性皱纹的疗效及可行性分析

目的:探讨A型肉毒毒素改善面上部动力性皱纹的临床疗效,并分析其可行性。方法:选择2016年2月到2016年10月在我院接受面上部动力性皱纹改善治疗的患者35例作为研究对象,所有患者均给予注射A型肉毒毒素治疗。评价并比较患者在治疗后30 min、治疗后3 d、治疗后7 d的优良率。采用面部皱纹量表(FWS)评价患者面部皱纹严重程度指数,比较患者治疗前、治疗后4周及治疗后6月的FWS指数。观察并比较治疗后面上部紧绷感、乏力感、注射点局部瘀斑等不良反应发生情况。结果:患者治疗后30 min、3 d及7 d时的优良率逐渐升高,治疗后3 d、7 d的优良率明显高于治疗后30 min,治疗后7 d的优良率明显高于治疗后3 d,差异均具有统计学意义(P0.05)。患者治疗后4周和治疗后6月的FWS指数均明显小于治疗前,并且治疗后4周的FWS指数明显小于治疗后6月,差异均具有统计学意义(P0.05)。治疗后,仅有4例患者出现轻微不适,所有症状均于休息3-4 d后自行消失,不影响患者的正常工作和生活。结论:A型肉毒毒素改善面上部动力性皱纹的临床疗效显著,维持时间较长,不良反应较少且较轻,安全性较好,值得在临床上推广应用。     

Effect of various upper limb multibody models on soft tissue artefact correction: A case study

Soft tissue artefacts (STA) introduce errors in joint kinematics when using cutaneous markers, especially on the scapula. Both segmental optimisation and multibody kinematics optimisation (MKO) algorithms have been developed to improve kinematics estimates. MKO based on a chain model with joint constraints avoids apparent joint dislocation but is sensitive to the biofidelity of chosen joint constraints. Since no recommendation exists for the scapula, our objective was to determine the best models to accurately estimate its kinematics. One participant was equipped with skin markers and with an intracortical pin screwed in the scapula. Segmental optimisation and MKO for 24-chain models (including four variations of the scapulothoracic joint) were compared against the pin-derived kinematics using root mean square error (RMSE) on Cardan angles. Segmental optimisation led to an accurate scapula kinematics (1.1° ≤ RMSE ≤ 3.3°) even for high arm elevation angles. When MKO was applied, no clinically significant difference was found between the different scapulothoracic models (0.9° ≤ RMSE ≤ 4.1°) except when a free scapulothoracic joint was modelled (1.9° ≤ RMSE ≤ 9.6°). To conclude, using MKO as a STA correction method was not more accurate than segmental optimisation for estimating scapula kinematics.     

Quantitative assessment of upper limb muscle fatigue depending on the conditions of repetitive task load

The aim of this study was to discriminate fatigue of upper limb muscles depending on the external load, through the development and analysis of a muscle fatigue index. Muscle fatigue is expressed by a fatigue index based on an amplitude parameter (calculated in the time domain) and a fatigue index based on a frequency parameter (a parameter calculated in the frequency domain). The fatigue index involves a regression function that describes changes in the EMG signal parameter, time elapsing before muscle fatigue and the probability of specific trends in changes in EMG parameters for the population under study.

The experimental study covered a group of 10 young men. During the study, they exerted force at a specific level and for a specific time in 12 load variants. During the study, EMG signals from four muscles of the upper limb were recorded (trapezius pars descendents, biceps brachii caput breve, extensor carpi radialis brevis, flexor carpi ulnaris). For each variant and for each examined muscles, the value of the fatigue index was calculated. Values of that index quantitatively expressed fatigue of a specific muscle in a specific load variant.

A statistical analysis indicated variation in the fatigue of the biceps brachii caput breve, extensor carpi radialis brevis, and flexor carpi ulnaris muscles depending on the external load (load variant) according to the task performed with the upper limb.

The study demonstrated usefulness of the fatigue index in expressing quantitatively muscle fatigue and in discriminating muscle fatigue depending on the external load.     

Kinematic models of the upper limb joints for multibody kinematics optimisation: An overview

Soft tissue artefact (STA), i.e. the motion of the skin, fat and muscles gliding on the underlying bone, may lead to a marker position error reaching up to 8.7 cm for the particular case of the scapula. Multibody kinematics optimisation (MKO) is one of the most efficient approaches used to reduce STA. It consists in minimising the distance between the positions of experimental markers on a subject skin and the simulated positions of the same markers embedded on a kinematic model. However, the efficiency of MKO directly relies on the chosen kinematic model. This paper proposes an overview of the different upper limb models available in the literature and a discussion about their applicability to MKO.The advantages of each joint model with respect to its biofidelity to functional anatomy are detailed both for the shoulder and the forearm areas. Models capabilities of personalisation and of adaptation to pathological cases are also discussed. Concerning model efficiency in terms of STA reduction in MKO algorithms, a lack of quantitative assessment in the literature is noted. In priority, future studies should concern the evaluation and quantification of STA reduction depending on upper limb joint constraints.     

Progress in applying the Three Rs to the potency testing of Botulinum toxin type A

Botulinum toxin type A (BTA) is being increasingly used for a range of therapeutic purposes and also for cosmetic reasons. For many years, the potency of BTA has been measured by using an LD50 assay in mice. This assay is a cause for concern due to its unpleasant nature and extreme severity, and the requirement for high numbers of mice to be used. Alternatives to this potency assay are presently reviewed with particular reference to the work at the National Institute for Biological Standards and Control (NIBSC), and to recent work by the UK manufacturer of the substance. An in vivo local paralysis assay with considerably less severity has been developed and is in use at the NIBSC. Alternative, ex vivo functional assays in use include the measurement of BTA-induced paralysis of neurally-stimulated rodent diaphragm or rat intercostal muscle. The latter method has the advantage of allowing more preparations to be derived from one animal. However, these ex vivo methods have not yet been fully validated and accepted by regulatory agencies as potency assays. Endopeptidase assays, although not measuring muscle paralysis directly, may provide a very useful consistency test for batch release and may replace the routine use of the LD50 test for that purpose. These assays measure the cleavage of the SNAP-25 protein (the final stage of BTA action), and have been validated for batch release by the National Control Laboratory (NIBSC), and are in regular use there. ELISA assays, used alongside the endopeptidase assay, also provide useful confirmatory information on the amounts of functional (and non-functional) BTA present. The UK manufacturer is further validating its endopeptidase assay, an ex vivo muscle assay and an ELISA. It is anticipated that their work will lead to a change in the product license, hopefully within the next two years, and will form a critical milestone towards the end of the LD50 potency test.     

A new child-friendly 3D bimanual protocol to assess upper limb movement in children with unilateral cerebral palsy: Development and validation

Unilateral cerebral palsy (uCP) causes upper limb movement disorders that impact on daily activities, especially in bimanual condition. However, a few studies have proposed bimanual tasks for 3D motion analysis. The aim of this study was to validate the new version of a child-friendly, 3D, bimanual protocol for the measurement of joint angles and movement quality variables. Twenty children with uCP and 20 typically developing children (TDC) performed the five-task protocol integrated into a game scenario. Each task specifically targeted one or two upper limb degrees of freedom. Joint angles, smoothness and trajectory straightness were calculated. Elbow extension, supination, wrist extension and adduction amplitudes were reduced; hand trajectories were less smooth and straight in children with uCP compared to TDC. Correlations between the performance-based score and kinematic variables were strong. High within and between-session reliability was found for most joint angle variables and lower reliability was found for smoothness and straightness in most tasks. The results therefore demonstrated the validity and reliability of the new protocol for the objective assessment of bimanual function in children with uCP. The evaluation of both joint angles and movement quality variables should increase understanding of pathological movement patterns and help clinicians to optimize treatment.ClinicalTrials.gov identifier: NCT03888443.