Temporal relationship of muscle weakness and pain reduction in subjects treated with botulinum toxin A.

Botulinum toxin A has demonstrated efficacy in relieving pain in spastic and nonspastic muscle conditions. This analgesic property has generally been attributed to muscular relaxation. This study demonstrates initial muscular relaxation and concomitant pain relief in a masticatory muscle model. However, as muscle power returns to normal, pain relief is still very evident. This result suggests that the analgesic effect attributed to botulinum toxin is more complex than simple muscular relaxation.     

In vivo targeting of DC-SIGN-positive antigen-presenting cells in a nonhuman primate model

In vivo targeting of antigen-presenting cells (APCs) with antigens coupled to antibodies directed against APC-specific endocytic receptors is a simple and a promising approach to induce or modulate immune responses against those antigens. In a recent in vitro study, we have shown that targeting of APCs with an antigen coupled to an antibody directed against the endocytic receptor DC-SIGN effectively induces a specific immune response against that antigen. The aim of the present study was to determine the ability of the murine antihuman DC-SIGN antibody AZN-D1 to target APCs in a cynomolgus macaque model after its administration in vivo. Immunohistochemical analysis demonstrated that macaques injected intravenously with AZN-D1 have AZN-D1-targeted APCs in all lymph nodes (LNs) tested and in the liver. DC-SIGN-positive cells were mainly located in the medullary sinuses of the LNs and in the hepatic sinusoids in the liver. No unlabeled DC-SIGN molecules were found in the LN of AZN-D1-injected macaques. Morphologic criteria and staining of sequential LN sections with a panel of antibodies indicated that the DC-SIGN-targeted cells belong to the myeloid lineage of APCs. In conclusion, this is the first study that shows specific targeting of APCs in vivo by using antibodies directed against DC-SIGN.     

Time course of recovery of juvenile skeletal muscle after botulinum toxin A injection: an animal model study

OBJECTIVE: To study the effects of neuromuscular blockade using botulinum toxin A on juvenile muscles at a dosage of 6 units/kg body weight in a rat model. DESIGN: A total of 34 male Sprague-Dawley rats (1-mo old) were used. A small incision was made along the posterior aspect of the left hind leg with the exposure of the gastrocnemius. Botulinum toxin A was injected at a dosage of 6 units/kg body weight in the medial and lateral heads of the muscle. An equivalent volume of saline were injected into the right gastrocnemius (control). Motor evoked action potentials, muscle force generation, and muscle mass and neuromuscular junction morphometry were analyzed at different time intervals up to 1 yr after toxin injection. RESULTS: During the 1-2 wks after botulinum toxin A injection, muscle mass, electrophysiologic variables, and muscle force generation were significantly reduced but returned to nearly normal at 6 mos postinjection. In the study group, neuromuscular junction gutter depth became significantly shallower 2 mos after injection, then normalized at 1 yr. There was a nonsignificant trend toward larger neuromuscular junctions from the gastrocnemius injected with botulinum toxin A. CONCLUSION: Our findings provide scientific evidence to support the clinical situation in which the interinjection interval of 3-6 mos of botulinum toxin A at a similar dosage is used.     

In vivo selective expansion of gene-modified hematopoietic cells in a nonhuman primate model

A major problem limiting hematopoietic stem cell (HSC) gene therapy is the low efficiency of gene transfer into human HSCs using retroviral vectors. Strategies, which would allow in vivo expansion of gene-modified hematopoietic cells, could circumvent the problem. To this end, we developed a selective amplifier gene (SAG) consisting of a chimeric gene composed of the granulocyte colony-stimulating factor (G-CSF) receptor gene and the estrogen receptor gene hormone-binding domain. We have previously demonstrated that primary bone marrow progenitor cells transduced with the SAG could be expanded in response to estrogen in vitro. In the present study, we evaluated the efficacy of the SAG in the setting of a clinically applicable cynomolgus monkey transplantation protocol. Cynomolgus bone marrow CD34(+) cells were transduced with retroviral vectors encoding the SAG and reinfused into each myeloablated monkey. Three of the six monkeys that received SAG transduced HSCs showed an increase in the levels of circulating progeny containing the provirus in vivo following administration of estrogen or tamoxifen without any serious adverse effects. In one monkey examined in detail, transduced hematopoietic progenitor cells were increased by several-fold (from 5% to 30%). Retroviral integration site analysis revealed that this observed increase was polyclonal and no outgrowth of a dominant single clonal population was observed. These results demonstrate that the inclusion of our SAG in the retroviral construct allows selective in vivo expansion of genetically modified cells by a non-toxic hormone treatment.     

肉毒毒素A对肌肉痉挛患者功能康复的作用

目的：探讨A型肉毒毒素（botulinum toxin type A，BTX—A）对上运动神经元损伤后肢体肌肉痉挛的治疗价值及其剂量影响。方法：选择48例上运动神经元损伤患者采用肉毒毒素A电刺激引导下局部肌肉注射治疗肌痉挛，其中把小腿三头肌、肱二头肌、屈指肌随机分成高低两个剂量组，观察剂量不同对疗效的影响，同时对所有患者制订注射后的目标，观察其达标情况。结果：肉毒毒素A注射后肌肉张力明显降低（P〈0．05），但在小腿三头肌、肱二头肌及屈指肌群中均未发现明显的量效关系，各配对大小剂量组肌张力评分差异无显著性意义（P〉0．05）；患者功能显著改善，康复目标总达标率为70．4％，肉毒毒素对上肢的粗大运动及下肢的步行功能的改善效果明显，而对手的精细活动功能的恢复效果欠佳。结论：肉毒毒素A对缓解上运动神经元损伤后的肢体肌肉痉挛，提高其生活自理能力及运动功能疗效显著，肉毒毒素作用的量效关系尚有待确认.     

A型肉毒毒素对大鼠离体十二指肠平滑肌自发性收缩的影响

目的 观察A型肉毒毒素(botulinum toxin A,BTX-A)对离体十二指肠平滑肌的自发性收缩是否存在抑制作用,这种抑制作用的时效特点与胆碱能M受体抑制剂作用的异同,BTX-A是否可抑制外源性乙酰胆碱(acetylcholine,ACh)增强的十二指肠收缩,BTX-A预处理平滑肌后外原性ACh是否可增强十二指肠平滑肌的收缩.旨在为临床应用BTX-A治疗小肠收缩功能紊乱所致的疾病提供理论和实验依据.方法 选取Sprague-Dawley大鼠,实验时击头部致昏后,距幽门0.5 cm处取1.0～1.5 cm的肠管,置于37℃Krebs液的恒温平滑肌槽中,肌条的一端固定在肌槽底部的塑料弯钩上,另一端固定在张力传感器上.肌条在1 g的前负荷下孵育,随机分为BTX-A组(n=12),阿托品组(n=12),ACh+ BTX-A素组(n=12),ACh+阿托品组(n=12),BTX-A+ ACh组(n=12).在自发性收缩平稳20分钟后,根据研究方案,分别加入BTX-A( 10 U/ml)或阿托品(1μmol/L)、ACh(100 μmol/L).Biolap 420 E生物机能实验系统记录十二指肠纵形平滑肌条在不同给药条件下的收缩变化.结果 BTX-A抑制了十二指肠平滑肌自发性收缩的频率、张力及振幅(P＜0.01),这种抑制作用持续＞1小时.阿托品抑制了十二指肠平滑肌自发性收缩的频率、张力及振幅(P ＜0.01),但用药5分钟后,十二指肠平滑肌的收缩开始恢复,用药10分钟后,十二指肠平滑肌的收缩已基本恢复;十二指肠平滑肌自发性收缩的频率、张力及振幅与用药前比较,差异无统计学意义(P＞0.05).ACh增强了十二指肠平滑肌自发性收缩的频率、张力及振幅(P＜0.01),BTX-A抑制了ACh增强的十二指肠平滑肌收缩的频率、张力及振幅(P ＜0.01).ACh增强了十二指肠自发性收缩的频率、张力及振幅(P＜0.01),阿托品抑制了ACh增强十二指肠平滑肌收缩的频率、张力及振幅(P＜0.01).BTX-A抑制了十二指肠平滑肌自发性收缩的频率、张力(P＜0.01)及振幅(P ＜0.05),加入外源性ACh后不能增强十二指肠平滑肌收缩的频率、张力及振幅.结论 BTX-A抑制离体十二指肠平滑肌自发性收缩,BTX-A的抑制作用与阿托品不同,表现出不完全抑制十二指肠的收缩频率、张力和收缩幅度,这种作用为逐渐而持续性抑制.BTX-A抑制了外源性ACh增强的十二指肠平滑肌的收缩.BTX-A抑制了十二指肠平滑肌自发性收缩后,外源性ACh不能增强十二指肠平滑肌的收缩,提示BTX-A可作用于突触后膜M受体,从而抑制了ACh与M受体结合,具有类似“阿托品样效应”.     

肉毒毒素临床应用进展及适应症审批的意义

肉毒毒素是肉毒梭菌增殖过程中产生的一种细菌外毒素,其可作用于周围神经末梢的神经肌肉接头处抑制突触前神经递质乙酰胆碱的释放,实现肌肉化学性去神经支配、镇痛、调节自主神经紊乱等效应.本文重点概述肉毒毒素的治疗效应,以及肉毒毒素临床应用进展和肉毒毒素适应症开发的临床价值.     

特发性脊柱侧凸的病因学研究:鸡脊柱侧凸模型椎旁肌中肌梭的形态学特征

目的观察鸡脊柱侧凸模型椎旁肌中肌梭结构、数量、密度及其分布,为特发性脊柱侧凸的病因提供理论依据. 方法对刚孵育出的鸡行松果体切除,获得脊柱侧凸动物模型;将鸡脊柱侧凸模型分为 Cobb角 10°～ 20°组及 Cobb角 >40°组,椎旁肌取下后切片,行 Masson三色染色,观察肌梭结构,对肌梭数量、密度进行统计学分析. 结果鸡脊柱侧凸模型椎旁肌中部分肌梭被囊呈"气球样变",梭内肌纤维有异常改变; Cobb角 10°～ 20°组肌梭密度为( 25± 5)个 /g肌肉. Cobb角 >40°组肌梭密度为( 23± 7)个 /g肌肉. 结论与正常对照组相比,鸡脊柱侧凸模型椎旁肌中部分肌梭结构异常,肌梭数量减少,密度降低.     

In vitro and in vivo effects of leukotriene B4 antagonism in a primate model of asthma.

To test the hypothesis that leukotriene (LT) B4 antagonists may be clinically useful in the treatment of asthma, CP-105,696 was evaluated in vitro, using chemotaxis and flow cytometry assays, and in vivo, using a primate asthma model. CP-105,696 inhibited LTB4-mediated monkey neutrophil chemotaxis (isolated cells, LTB4 = 5 nM) and CD11b upregulation (whole blood, LTB4 = 100 nM) with IC50 values of 20 nM and 16.5 microM, respectively. Using a modification of a previously described in vivo protocol (Turner et al. Am. J. Respir. Crit. Care Med. 1994. 149: 1153-1159), we observed that treatment with CP-105,696 inhibited the acute increase in bronchoalveolar lavage (BAL) levels of IL-6 and IL-8 by 56.9 +/- 13.2% and 46.9 +/- 14.5%, respectively, 4 h after challenge with Ascaris suum antigen (Ag). CP-105,696 tended to reduce the increase in BAL protein levels 0.5 h after Ag challenge by 47.5 +/- 18.3%, but this was not statistically significant. In addition, CP-105,696 prevented the significant 11-fold increase in airway responsiveness to methacholine after multiple Ag challenge. These results suggest that LTB4 partially mediates acute and chronic responses to antigen in an experimental primate asthma model and support the clinical evaluation of LTB4 antagonists in human asthma.     

肉毒毒素A作用下肌细胞形态的变化规律研究

目的探讨肉毒毒素A(BTX-A)作用下肌细胞形态的变化规律。方法 90只成年雄性SD大鼠通过随机数字表随机分为实验组和对照组,每组45只,分别注射BTX-A与生理盐水,每组通过随机数字表法又分为9个时间组,每时间组5只大鼠,按观察时间的不同(注射后30分钟、6小时、1天、3天、1周、2周、4周、6周和8周),采用线粒体酶组织化学染色的方法动态观察局部注射BTX-A后大鼠胸舌骨肌纤维直径变化。结果注`射BTX-A后30分钟,肌纤维直径开始变小,萎缩于2~4周内接近高峰,大多数肌纤维的这种萎缩状态会持续到8周以后;从第4周起,点状区域的肌纤维直径有恢复的趋势,而且这些肌纤维的酶活性也比其它肌纤维高;注射BTX-A后白肌纤维受影响最早而且明显,恢复亦最慢,红肌纤维受影响迟而恢复快。结论注射BTX-A后,大鼠胸舌骨肌的线粒体酶活性受到抑制,能量代谢出现障碍,肌纤维随之发生去神经性萎缩;但随着神经发芽和新轴突生成,BTX-A导致的骨骼肌能量代谢障碍及萎缩也逐渐恢复。     

Effects of intra-articular botulinum toxin type A in an equine model of acute synovitis: a pilot study

OBJECTIVE: To test the hypothesis that botulinum toxin type A (BoNT-A) can attenuate lameness associated with acute synovitis in an equine model. DESIGN: Four horses 2-6 yrs of age with clinically normal carpi were studied for 15 days. Kinematic gait analysis and clinical measures of lameness were conducted before and after experimental interventions. Horses were randomly assigned to either placebo (saline) or treatment (BoNT-A) groups. On day 0 of the intervention, 50 units of BoNT-A or an equivalent volume of saline (0.09%) was given into the middle carpal joints. On day 14, acute synovitis was induced with intra-articular injection of recombinant equine interleukin-1 beta (IL-1 beta) 100 ng. Synovial fluid, clinical evaluation of lameness, and kinematic gait analysis were evaluated on day 15. RESULTS: Synovitis was observed on histology and cytology in all horses after IL-1 beta, indicating acute suppurative inflammation. In the BoNT-A group, one horse developed lameness, whereas the other demonstrated no change in baseline gait evaluation. No adverse effects were observed in joints injected with BoNT-A or with saline alone. CONCLUSIONS: Our findings support the idea that BoNT-A can attenuate lameness in an equine model of acute synovitis. Our findings further suggest that BoNT-A might be a potential new treatment for painful arthritis; this warrants further study.     

A型肉毒毒素关节腔注射对佐剂性关节炎大鼠的抗炎效果观察

目的观察A型肉毒毒素(BoNT/A）踝关节腔注射对佐剂关节炎大鼠模型的抗炎作用。 方法选取Wistar大鼠90只,于左后足趾皮下注射完全弗氏佐剂（CFA）0.1ml进行免疫,从而制作大鼠佐剂诱导性关节炎(AIA)动物模型。待AIA模型成模后,采用随机数字表法将其分为BoNT组(向踝关节腔内注射BoNT/A)、生理盐水组(向踝关节腔内注射生理盐水)和假手术组（行踝关节腔穿刺但不注射药物）;另选取30只健康Wistar大鼠纳入空白组。大鼠于免疫后每隔3d行踝关节红外热成像拍摄以及踝关节关节炎指数评分;成模后分别于成模当天、成模后7d、成模后14d时行红外热成像拍摄、白细胞介素-1β（IL-1β）蛋白检测以及踝关节HE染色、评分。 结果大鼠免疫后3d时其关节炎指数开始升高,于免疫10d左右时关节炎指数明显升高,并随疾病进展逐渐增高,至免疫18~20d时达到峰值。红外热成像检查显示,大鼠免疫后出现右侧踝关节区域温度明显升高,随着炎症发展,温度呈逐渐下降趋势,免疫20d时双侧踝关节平均温度维持在37.5~38.0℃。成模后7d、14d时BoNT组可观察到双侧足趾平均温度较成模当天时明显下降,与同时间点生理盐水组及假手术组间差异均具有统计学意义（P<0.05）;成模后7d、14d时BoNT组踝关节腔滑膜中IL-1β蛋白表达较成模当天时明显降低,并且与相同时间点生理盐水组及假手术组间差异亦具有统计学意义（P<0.05）。成模后7d、14d时除空白组外,其余各组大鼠滑膜组织增生、炎性细胞浸润、软骨破坏及软骨下骨质暴露程度均较成模当天时有不同程度加重;并且BoNT组在成模后7d、14d时HE评分［分别为（3.8±0.63）分和（3.7±0.86）分］均显著低于相同时间点模型组及生理盐水组水平(P<0.05)。 结论关节腔内注射BoNT/A对佐剂关节炎大鼠具有抗炎作用。     

A pilot study of botulinum toxin A for headache in cervical dystonia

OBJECTIVE: To evaluate the prevalence of associated headache (HA) pain with craniocervical dystonia and the therapeutic effect of BoNT-A injections on the HA component when injected for cervical dystonia. BACKGROUND: HA associated with craniocervical dystonia is a recent formally codified entity, but has not been systematically studied. METHODS: We identified 44 subjects from three movement disorder clinics who presented with craniocervical dystonia and concurrent HA pain. The subjects were injected with botulinum toxin type A (BoNT-A) and prospectively evaluated with the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS), headache diaries, Headache Impact Test (HIT-6), and Migraine Disability Assessment Scale (MIDAS), along with HA pain anatomy and adverse events, at baseline, and at 4, 8, and 12 weeks post-injection. RESULTS: As expected, all aspects of the TWSTRS robustly improved. Headache diaries and the HIT-6 also improved at 4, 8, and 12 weeks post-injection. Sections of the MIDAS improved, and adverse events were minimal. CONCLUSION: BoNT-A safely improves headache associated with craniocervical dystonia when administered for the primary condition of craniocervical dystonia.     

Effect of botulinum toxin on endplate noise in myofascial trigger spots of rabbit skeletal muscle

OBJECTIVE: To assess the effect of botulinum toxin type A (BTX-A) on the endplate noise prevalence in rabbit myofascial trigger spots to confirm the role of excessive acetylcholine release on the pathogenesis of myofascial trigger points and to develop an objective indicator of the effectiveness of BTX-A in the treatment of myofascial trigger points. DESIGN: Eighteen adult New Zealand rabbits were divided into three groups that received a single bolus of BTX-A over a myofascial trigger spot region on one side of the biceps femoris muscle. Another 10 rabbits received multiple-point injections in a myofascial trigger spot where endplate noises were found. A control study was performed on the other side of the biceps femoris muscle. The endplate noise prevalence in a myofascial trigger spot region was assessed. RESULTS: It was found that injection of BTX-A reduced the prevalence of endplate noise. No significant differences between a single bolus injection and multiple-point injections were noted, although there was some evidence that multiple-point injections might maintain the endplate noise decreasing effect much longer than a single injection. CONCLUSIONS: This study demonstrated the suppressive effect of BTX-A on endplate noise prevalence in a myofascial trigger spot region. The prevalence of endplate noise in the myofascial trigger point region may be a useful objective indicator for evaluating the therapeutic effectiveness of BTX-A injection to treat myofascial trigger points.     

Parent evaluation of spasticity treatment in cerebral palsy using botulinum toxin type A

OBJECTIVE: To assess parent ratings of treatment acceptability associated with botulinum toxin type A (BTX-A) injection for spasticity in children with cerebral palsy (CP). DESIGN: A single-point survey design across a sequentially recruited cohort, using a standardized evaluation measure. SETTING: Regional specialty health care center medical clinic and pain research program. PARTICIPANTS: Fifty-nine parents of children with CP receiving BTX-A injection for spasticity. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Treatment Evaluation Inventory. RESULTS: Overall, parent ratings of treatment acceptability ranged from moderate to high. There were no significant differences for caregiver ratings in relation to characteristics of the raters (age, sex, marital status) or of the children (age, sex, mental retardation, severity of disability) characteristics. CONCLUSIONS: These findings indicate that on average, parents of children with CP consider BTX-A treatment for the management of spasticity to be an acceptable form of treatment.     

Postpoliomyelitis muscle weakness: a prospective study of quadriceps strength

The objective of this study was to evaluate the presence of progressive postpoliomyelitis muscle weakness (PPMW) in affected individuals 20 to 40 years after the initial polio infection. Over a three-year period, the isometric and isokinetic strength of the quadriceps femoris muscle was studied in seven symptomatic patients with previous poliomyelitis (mean = 38.3 years from infection) to determine if quadriceps strength decreased during the three years. Each patient had a quadriceps affected by polio on one side and a clinically nonaffected quadriceps on the contralateral limb. The maximal isometric force and the peak isokinetic force of the affected quadriceps (AQ) and nonaffected quadriceps (NQ) muscles were tested on a computerized isokinetic dynamometer machine at six-month intervals. Isometric force increased significantly, by 29% per year (p less than .02) in the AQ and by 14% per year (p less than .05) in the NQ. Paired analysis to determine the change in strength between the affected and nonaffected muscles for the isometric data showed a mean nonsignificant increase in the AQ of 14% per year (p = .01). The change in peak isokinetic force demonstrated a significant increase in the AQ of 35% per year (p less than .05); whereas, the NQ peak isokinetic force increased 15% per year which was not statistically significant. Paired analysis to determine the change in strength between the affected and nonaffected muscles for the isokinetic data showed a nonsignificant relative increase in the AQ of 20% per year (p less than .06).(ABSTRACT TRUNCATED AT 250 WORDS)     

The role of botulinum toxin injections in the management of muscle overactivity of the lower limb

Muscle overactivity is common in patients with adult onset central nervous system damage. It can produce significant disablement in conjunction with other impairments such as adaptive soft tissue shortening and loss of muscle strength. Muscle overactivity is not evenly distributed throughout the body; across joints there is frequently imbalance between agonist and antagonist, producing abnormal joint postures and movement patterns. Due to the asymmetric nature of the abnormal activity across joints, in general we recommend local treatment targeting the more overactive of the two agonists, rather than systemic treatment. Considerable experience with the use of botulinum toxin, both serotypes A and B, in the treatment of muscle overactivity has been accumulated in the last two decades through pragmatic clinical practice and open label studies, supported by an increasing number of randomized controlled trials. In most cases, it is important to use botulinum toxin injection for treatment of muscle overactivity in the setting of wider rehabilitation goals and interventions. Focal and partial blocks with botulinum toxin should be used as a component of a general neurorehabilitation programme rather than as an alternative to other treatments. We review the evidence supporting the use of botulinum toxin to treat muscle overactivity in the lower limb, present practical guidelines on when and how to use botulinum toxin and provide direction for future research.     

超声引导下的肩胛下肌外侧（腋下）入路肉毒毒素注射

目的:介绍超声引导下肩胛下肌外侧腋下入路肉毒毒素注射方法。方法:2例上肢偏瘫肩痛并有痉挛的患者,采用超声引导下外侧腋下入路注射肉毒毒素。结果:经注射治疗后,患者肩痛、上肢痉挛明显降低;主动和被动肩关节活动度均有提高。结论:某些深部肌肉注射肉毒毒素具有一定技术难度和风险。由于在超声引导下,深部肌肉可视并容易注射,注射具有安全性、有效性和经济性。