不同延期神经再支配环杓后肌方法的疗效观察

目的：探讨环杓后肌延期神经再支配的方法及时机。方法：将２９只狗分成三组，两个实验组在喉返神经切断后即刻、４、６、８、１０、１２个月分别将膈神经与喉返神经吻合选择性支配环杓后肌和颈袢神经—胸骨甲状肌蒂植入环杓后肌，对照组喉返神经切断后各时间不作任何手术。结果：两个实验组６个月后行喉镜、神经肌电及肌收缩力、组织化学证实：神经吻合组各时间效果明显好于神经肌蒂植入组，但两种术式疗效均随病程延长而下降。结论：神经吻合法延期神经再支配环杓后肌６个月内效果较好，而神经肌蒂植入法治疗环杓后肌麻痹最好不晚于失神经后４个月，否则效果不佳     

环杓关节在延期喉内肌神经再支配后的变化

目的：观察环杓关节在延期喉内肌神经再支配后的变化及对杓状软骨运动的影响。方法：在喉返神经切断后，将犬分为两组：实验组（６、８、１０、１２和１８个月后行延期神经再支配）和对照组（相同时间无神经再支配），观察其杓状软骨运动及环杓关节组织学变化。结果：１２个月以内行喉内肌神经再支配后环杓关节纤维化有不同程度的逆转，而１８个月行喉内肌神经再支配后环杓关节纤维化不可逆。结论：越早行喉内肌神经再支配对环杓关节     

不同延期神经再支配环杓的肌方法的疗效观察

探讨环杓后肌延期神经再支配的方法及时机。方法：将２９只狗分成三组，两个实验组在喉返神经切断后即刻、４、６、８、１０、１２个月分别将膈神经与喉返神经吻合选择性支配环杓后肌和颈袢神经一胸骨甲状肌蒂植入环杓后肌。     

失神经支配后喉内肌细胞增殖状态的变化

目的 了解犬喉内肌失神经支配后细胞萎缩和增殖状态的变化规律，对反映肌肉不可逆发迹的指标进行初步探讨。方法 在喉返神经切断后不同时间，取犬的甲杓肌及环杓侧进行如下观察：（１）ＨＥ染色测肌细胞的截面积；２）ＶａｎＧｉｅｓｏｎ染色测定胶原纤维与肌细胞面积比；（３）ＣＡＳ２００图像分析系统ＤＮＡ定量测定方法统计Ｓ期细胞核比值。结果 甲杓肌及环杓侧肌失神经支配后６个月内肌细胞的截面积无明显改变，６个月后肌我     

甲杓肌颈袢主支植入术后收缩特性

将７条犬的左喉返神经、喉上神经外支切断后，以左颈袢主支植入左声带内收肌中，６个月后用张力换能器检测。结果７条犬术侧甲杓肌均有收缩反应，收缩强度为正常侧的３８％～９２％，最大单收缩时间较正常侧延长１３％～３６％。提示肌肉收缩特性的研究能较准确地反映再神经支配后肌功能恢复情况。     

一侧膈神经上根选择性神经再支配环杓后肌的实验研究

目的 探讨一侧膈神经上根选择性神经再支配环杓后肌的可行性和有效性.方法 8只健康雄性青年Beagle犬作为实验动物,全麻下切断Beagle犬左侧膈神经上根与左侧喉返神经,两断端通过游离神经桥接的方式吻合,并将同侧喉返神经内收肌支切断并植入同侧环杓后肌中;右侧不做任何处理,作为正常对照侧.于手术前、神经修复术后即刻、术后6个月分别行电子喉镜和喉肌电图检查,在最后一次检查后处死动物,取双侧环杓后肌和喉内段喉返神经行组织学检查并与对侧比较.结果 术前8只犬的双侧声带运动正常,术后即刻左侧声带固定,术后6个月均恢复了吸气性外展运动.术前8只犬双侧环杓后肌均能记录到自发肌电位,并引出诱发电位;术后即刻左侧环杓后肌呈电静息,未记录到诱发电位;术后6个月均能记录到自发肌电位,并引出诱发电位,而且两种电位幅度分别与术前比较差异均无统计学意义(均为P>0.05).环杓后肌Masson染色示两侧肌肉纤维相对截面积、胶原纤维相对截面积、肌肉/胶原纤维截面积比差异均无统计学意义(均为P>0.05).喉内段喉返神经甲苯胺蓝染色后示神经纤维分布较均匀密集,且左右两侧喉返神经有髓神经纤维数量差异无统计学意义(P>0.05).结论 左侧膈神经上根选择性神经再支配环杓后肌能有效避免神经错向再生,恢复声带的生理性外展运动.     

环杓后肌失神经及神经再支配后兴奋性的研究

为研究环杓后肌失神经及其神经再支配后的兴奋性,将15只狗分成3组即神经植入组、神经肌蒂组及对照组,每组5只.术后6个月发现植入神经、带肌蒂的神经均与正常喉返神经兴奋性无显著性差异(P>0.05),且神经兴奋性高于肌肉兴奋性.两个恢复神经再支配组的环杓后肌与正常环杓后肌兴奋性亦无显著性差异(P>0.05),而失神经的环杓后肌与两个神经修复组及正常环杓后肌的兴奋性则有高度显著性差异(P<0.01),且其兴奋性最低.结果表明,当功能性电刺激治疗双侧喉返神经麻痹时,刺激电极安放在修复的神经上比安放在环杓后肌处效果要好.     

实验性声带麻痹神经吻合术后神经传导初步研究

目的研究喉返神经完全性损伤后颈袢神经支单纯支配环杓侧肌的电生理恢复情况。方法将实验组犬左喉返神经完全切断,3个月后,再将其内收肌支与同侧颈袢胸舌肌支延期吻合,吻合术后6个月检测环杓侧肌肌电活动情况。结果吻合术后6个月,左环杓侧肌可见肌电动作电位,其潜伏期为1.60±0.05ms,与术前相比较无显著差异,但神经传导速度为15.60±0.25m/s,较术前减慢,差异有显著性。结论喉返神经损伤后3月再行选择性环杓侧肌神经支配吻合术,能基本恢复肌肉电生理功能,但要达到完全正常水平,仍有待继续观察。     

用颈交感干和舌下神经袢再支配狗环杓后肌的研究

将8只狗的右侧喉返神经及其内收支切断，再分别将其中4只狗的右侧颈交感神经干和2只狗的右侧舌下神经袢切断后，各将其近侧端和已切断的喉返神经的远侧端吻合，其余2只狗作为失神经支配的对照组。分别于术前、术后即刻及术后4个月行声门照像、肌电图观察，最后行环杓后肌肉眼及组织化学观察。实验结果证明：采用两种神经修复喉返神经均可部分恢复环杓后肌功能，论证了用颈交感神经干修复的机理及各种神经修复法的优缺点，为今后进一步的研究和临床应用提供理论依据。     

实验性狗环杓后肌神经移植后的功能效果

本研究旨在比较一组巳行喉返神经切断和各种类型环杓后肌神经移植的狗所取得的功能效果。作者行喉返神经端对端吻合,即神经自身吻合6例;支配胸骨甲状肌的颈神经袢与喉返神经吻合5例;膈神经与喉返神经吻合5例。除3例神经自身吻合外,所有实验狗的喉返神经内收肌支均予切断,结扎两断端。依神经再生距离的不同,于手术后3～4月(神经自身     

Study on contractile properties of the posterior cricoarytenoid muscle after delayed reinnervation]

To study date on the contractile properties of posterior cricoarytenoid muscle after delayed reinnervation of different reinnervated methods. Twenty four dogs were reinnervated at 0,4,5,6,10 and 12 month interval following recurrent laryngeal nerve via the phrenic nerve anastomosed to the recurrent laryngeal nerve after cutting the adductor branch and ansa cervicalis-sternothyroid muscle pedicle implanted into the posterior cricoarytenoid muscle. After 6 months, a series of contractions were recorded from each side in twenty living dogs. The results showed that contractile force of reinnervated muscle decreased gradually with the time of denervation, but contractile force of muscle was no significantly difference between reinnervated side of nerve anastomosed group in 4 months after denervated and normal side, and it was significantly difference between nerve anastomosed group and nerve-muscle pedicle implanted group at some time of delayed reinnervation. The contractile time of reinnervated side of two operated groups was similar to that of normal side. The conclusion demonstrated that the contractile properties can indicate exactly reinnervated degree of muscle, and the earlier reinnervation was performed, the better curative effect was.     

Intrinsic laryngeal muscle reinnervation using the muscle-nerve-muscle technique

OBJECTIVES: This study was performed to investigate the muscle-nerve-muscle reinnervation technique in the larynx, in which a nerve conduit implanted into an innervated muscle conducts axonal sprouting into a denervated muscle while maintaining function of the donor muscle. METHODS: In this study, the muscle-nerve-muscle technique was used to direct superior laryngeal nerve axons to reinnervate intrinsic laryngeal muscles by implanting the recurrent laryngeal nerve stump into the cricothyroid muscle in 8 dogs. In 4 of the dogs, the recurrent laryngeal nerve trunk to the adductor muscles was divided so that all axonal sprouting was directed to the posterior cricoarytenoid muscle. Six-month electromyography data were obtained from 6 of the 8 dogs. RESULTS: All 6 dogs showed evidence of successful reinnervation of the thyroarytenoid or posterior cricoarytenoid muscles with action potentials that corresponded to spontaneous respiratory efforts, while the donor cricothyroid muscles retained their phasic contraction. These responses were obliterated when the recurrent laryngeal nerve conduit was divided. Histologic examination of the intrinsic laryngeal muscles demonstrated successful reinnervation. CONCLUSIONS: The results confirm that intrinsic laryngeal muscles may be successfully reinnervated by the superior laryngeal nerve with the muscle-nerve-muscle technique, without sacrifice of function of the cricothyroid muscle. This method offers an alternative source of appropriately firing axons for laryngeal reinnervation procedures.     

膈神经替代喉返神经修复治疗双侧声带麻痹

目的 探讨膈神经喉返神经吻合和内收肌支环杓后肌植入术（膈神经手术）治疗双侧喉返神经损伤声带麻痹的有效性、可行性。方法 第二军医大学长海医院耳鼻咽喉科1999年8月－2001年7月治疗外伤性双侧喉返神经损伤声带麻痹6例。病程1周－18个月，一侧作膈神经手术，而另一侧作颈袢肌蒂环杓后肌植入术。手术前后电子喉镜、频闪喉镜观察声门大小、声珲运动、振动情况，噪音声学参数分析，喉肌电力产检查评价手术效果。结果 术后2－3周检查发现4例声门较术前增大2－3mm，但声带固定不动，2例无明显改善。术后6个月5例膈神经修复侧均恢复了较大幅度的吸气性声带外展功能，外展幅度可达3－5mm，而肌蒂植入侧仅轻微外展或固定不动，幅度均在1mm以内。此5例均顺利拔管，并能承受较大强度的体力活动，1例仍在随访中。术后4个月6例肌电图检查显示膈神经修复侧自发、诱发电位均明显大于肌蒂植入侧，自发电活动与肋间肌基本同步，而较肌蒂植入侧延迟100－200ms。声音估价显示3例声嘶术后较术前好转，2例无变化。术后半年肺功能均恢复正常。结论 膈神经喉返神经吻合内收肌支环杓后肌植入术安全可行，较颈袢肌蒂植入术更能有效地恢复声带吸气性外展运动，值得临床推广应用。     

喉返神经-膈神经吻合术后声带外展肌的神经肌电图表现

目的研究声带外展肌-环杓后肌在喉返神经-膈神经吻合术前、后的神经肌电图表现。方法造成犬单侧喉返神经完全性损伤,即行单侧犬喉返神经-膈神经吻合术,对吻合术前、后同侧环杓后肌神经电图各项参数进行对比分析。结果吻合术前环杓后肌的神经电图表现为潜伏期1．26±0．86,神经传导速度15．08±0．47ms／s,幅值50．49±4．75mV。神经吻合术后4个月,环杓后肌神经电图出现清晰动作电波形,表现为潜伏期延长,传导速度减慢,动作电位峰-峰值明显减小。结论神经吻合术后4个月,出现环杓后肌肌纤维不同程度的神经再支配,但新生神经末稍终板数量及神经鞘膜之再生还未完全达到正常水平。     

Experimental laryngeal reinnervation by phrenic nerve implantation into the posterior cricoarytenoid muscle

Under general anaesthesia, 5 dogs underwent sectioning of the right recurrent nerve followed by implantation of the phrenic nerve into the posterior cricoarytenoid (PCA) muscle. Some 6-7 months later the dogs were sacrificed after registration of vocal cord motility. Still photographs and movie film of the larynx were taken during quiet and forced respiration and at electrical stimulation of the implanted phrenic nerve. The PCA and vocal muscles were removed for histochemical studies. We found practically no abductory movement of the vocal cord on the reinnervated side, either during quiet or forced respiration. During forced inspiration there was, however, a slight medial bowing of the right vocal cord. At electrical stimulation there was a sphincteric movement of the entire larynx. Histochemistry showed a reinnervation picture of both the PCA and the vocal muscles on the experimental side. The conclusion drawn from this study is that axonal escape, probably from the implantation site, results in an unwanted reinnervation of laryngeal adductor muscles, which neutralize the abducting effect of the PCA muscle during inspiration. This method therefore does not seem to be suitable as a treatment alternative for bilateral recurrent nerve paralysis.     

家犬双侧环杓后肌失神经及颈袢神经再支配后肌卫星细胞活性变化

目的 探讨家犬双侧环杓后肌失神经支配及用颈袢神经再吻合支配后肌卫星细胞活性的变化.方法 随机数字表法将24只家犬分成3组,每组8只,分别为切断双侧喉返神经组,切断喉返神经后即刻颈袢神经再吻合组,不切断喉返神经的对照组.3组动物手术后再饲养9周后再次暴露喉返神经及环杓后肌,使用喉返神经诱发环杓后肌电位来验证喉返神经的再通情况;双盲法提取环杓后肌肌肉组织中总RNA,反转录后做实时定量聚合酶链反应(PCR)检测肌卫星细胞增殖分化标记物Myogenin、Myf5和Pax7的mRNA表达.结果 手术后3组动物各有1只死亡或感染退出实验.术后9周诱发神经环杓后肌肌电图提示对照组动物喉返神经功能均正常,切断组7只动物电刺激都没有反应,吻合组均有神经再通.Myogenin的mRNA相对表达量切断组较对照组和吻合组均有明显升高(Z值为1.42和1.38,P值均＜0.05),Myf5的mRNA表达切断组也明显高于对照组和吻合组(Z值为1.66和1.69,P值均＜0.01);切断组Pax7的mRNA表达明显高于对照组(Z=1.66,P＜0.01),也高于吻合组(Z=1.42,P＜0.05);而吻合组与对照组Myogenin、Myf5和Pax7的mRNA表达差异均无统计学意义(P值均＞0.05).结论 双侧喉返神经切断后,环杓后肌的肌卫星细胞增殖分化的mRNA表达增强,而同颈袢神经再吻合后肌卫星细胞的增殖分化表达下降.

Abstract：

Objective To investigate the activity of bilateral posterior cricoarytenoid muscle satellite cell after denervation or reinnervation with ansa cervicalis. Methods Twenty four dogs were randomly divided into 3 groups. The bilateral laryngeal recurrent nerves were cut in group one in all dogs. The bilateral laryngeal recurrent nerves were anastomosed with ansa cervicalis after incision in group two in all dogs. The dogs in group three were used as control. Nine weeks after surgery, the electromyography was used to test the regeneration of the nerve. The posterior cricoarytenoid muscles biopsy were collected. The expression of mRNA of Myogenin, Myf5, and Pax7 was assayed by realtime RT-PCR after total RNA isolation. Results Two dogs died after surgery in incision and anastomose group. The electromyography suggested that the RLN of all dogs had denervated in the incision group and had reinnervated in the anastomose group after 9 weeks. Myogenin mRNA from RLN incision dogs PCA muscles had greater expression versus controls ( Z = 1.42, P ＜ 0. 01 ) or anastomosed dogs ( Z = 1.38, P ＜ 0. 01 ). Myf5 mRNA expression from RLN incision dogs PCA muscles had significant increase versus control dogs ( Z = 1.66, P ＜0. 01 ) or anastomosed dogs ( Z = 1.69, P ＜ 0. 01 ). Pax7 mRNA expression from RNL incision dogs had significant increase compared with control ( Z = 1.66, P ＜0. 01 ) or anastomosed animals ( Z = 1.42, P ＜0. 05 ). There was no significant difference in Myogenin ( Z = 1.34, P ＞ 0. 05 ), Myf5 ( Z = 0. 54, P ＞0. 05) and Pax ( Z = 0. 54, P ＞ 0. 05 ) mRNA expression between controls and anastomosed animals.Conclusions The bilateral denervation of RLN cause significantly increasing in dog PCA muscle satellite cell proliferation and differentiation. The bilateral reinnervation of RLN cause PCA muscle satellite cell come back nonproliferative, quiescent state in dog.     

Experimental laryngeal reinnervation by phrenic nerve implantation into the posterior cricoarytenoid muscle

Under general anaesthesia, 5 dogs underwent sectioning of the right recurrent nerve followed by implantation of the phrenic nerve into the posterior cricoarytenoid (PCA) muscle. Some 6-7 months later the dogs were sacrificed after registration of vocal cord motility. Still photographs and movie film of the larynx were taken during quiet and forced respiration and at electrical stimulation of the implanted phrenic nerve. The PCA and vocal muscles were removed for histochemical studies. We found practically no abductory movement of the vocal cord on the reinnervated side, either during quiet or forced respiration. During forced inspiration there was, however, a slight medial bowing of the right vocal cord. At electrical stimulation there was a sphincteric movement of the entire larynx. Histochemistry showed a reinnervation picture of both the PCA and the vocal muscles on the experimental side. The conclusion drawn from this study is that axonal escape, probably from the implantation site, results in an unwanted reinnervation of laryngeal adductor muscles, which neutralize the abducting effect of the PCA muscle during inspiration. This method therefore does not seem to be suitable as a treatment alternative for bilateral recurrent nerve paralysis.