带蒂骨骼肌桥接近端周围神经防治残端痛性神经瘤的实验研究

目的 :探讨神经瘤的形成机制和致痛机理及周围神经与带蒂骨骼肌桥接吻合后神经瘤的生长情况。方法 :Wistar大白鼠 4 0只 ,随机分为实验组与对照组 ,两组均将左侧坐骨神经在适当水平切断 ,远端神经切除 ,实验组近端神经分为两束 ,用带蒂骨骼肌桥接两个神经断端 ,对照组近端神经置于原位。饲养 16周进行检测。结果 :实验组再生的神经纤维顺利通过吻合口长入骨骼肌肌桥内 ,并在肌桥的肌束内生长 ,分布于肌纤维之间 ,未见神经瘤形成 ;对照组均有典型的神经瘤形成。结论 :采用带蒂骨骼肌桥接断裂的周围神经近端后再生的神经纤维能顺利通过吻合 ,并沿肌外膜内和肌束之间生长 ,可防止神经瘤的形成。     

周围神经断端桥接后数量放大效应的研究

目的证实周围神经横截面不对等修复时神经纤维数量的放大效应。方法选用雄性Sprague-Dawley大鼠50只,分为A、B、C、D、E5组。A、B、C3组将大鼠坐骨神经切断后,A组近端保留完整断端与远端坐骨神经用甲壳质套管留置2mm间隙套接,B组在近端5mm处将坐骨神经中的腓总神经束结扎切断,将胫神经束与远端坐骨神经套接,C组在近端5mm处将坐骨神经中的胫神经束结扎切断,将腓总神经神经束与远端坐骨神经套接;D、E两组将胫神经在分叉处远端5mm组将胫神经切断;D组结扎切断2／3近端纤维,将剩余神经纤维与远端胫神经进行甲壳质套管套接,E组将全部近端纤维与远端胫神经进行甲壳质套管套接。1、2、4、12周后分别取材进行组织学和电生理研究。结果采用SPSS软件进行统计学分析。结果12周后电生理检查发现,各组诱发出的最大波幅下面积B、C两组小于A组（均P〈0．05）,D组小于E组（P〈0．05）。A组与B、C组之间,D、E组之间感觉神经传导速度相近。锇酸染色有髓神经纤维计数：各组远端均大于近端：A组远端比近端增加34．4％,B组增加39．6％,C组增加80．4％,D组增加101．1％,E组增加48．9％（P〈0．05）。结论在周围神经桥接后,远端神经纤维数量明显大于近端,存在神经纤维数量的放大;同源的神经桥接的放大效应大于非同源的神经。临床上较细神经修复远端粗大神经是可能的。     

HRP直接触涂周围神经断端追踪神经纤维来源的研究

<正> 自Kristensson等将辣根过氧化物酶(HRP)追踪神经联系的方法用于周围神经的研究以来,研究周围神经者,一般是将HRP液注射于末梢部位,或注射于神经干内,或将切断的神经近侧断端浸泡于HRP液中,而在相应的神经元胞体中显示出阳性反应产物。但注射于末梢部位者,HRP用量较大,注射于神经干内者,标记细胞的数量不如浸泡法的多,而浸泡法的浸泡时间常长达数小时,如果切断的神经近侧段部位太深、长度较短则操作起来很不     

Experimental studies on peripheral nerve regeneration enhanced by nerve growth factor

Summary The effect of exogenous nerve growth factor(NGF) examined on the neural repair of adult rabbit sciatic nerve was evaluated in the present study. A nerve regeneration chamber was created by suturing the proximal and distal ends of a transected sciatic nerve into a silicone chamber, A gap of 6 mm in chamber was left after removal of a 3mm piece of nerve in the distal ends and insertion of the proximal andl distal stumps into the chamber. Animals were operated on bilaterally, one side of the chamber was filled with a 1 mg/ml NGF/normal saline(NS) (experimental)and the contralateral side with NS(control). The regenerated nerves from within the silicone chamber were dissected and fixed 1 to 5 weeks following surgery for histological studies at both the light microscopic and ultrastructural levels. The NGF chamber showed a more mature regenerated nerve based on a larger diameter of the regenerated nerve trunk, a great number of axons, and thicker myelin sheaths. The project was supported by National Natural Science Foundation of China.     

Rapid immunostaining of live nerve for identification of sensory and motor fasciculi.

Six New Zealand white rabbits were anesthetized with pentobarbital, and sciatic nerves were exposed and cut at the mid thigh. Both proximal and distal ends were incubated directly with Blue-SAb in a micropipe of 5 mm diameter covering the nerve trunk ends for 30 minutes at room temperature. After removal of the micropipe, the nerve ends were washed with physiological saline. Blue-stained fasciculi, i.e., sensory fasciculi were seen among unstained ones under the operating microscope. This method requires no histological sections. Neural cells of the spinal cord and ganglion were cultured in RPM11640 medium containing Bright Blue. The growth and metabolism of the neural cells were tested by MTT assay and their morphology was observed. Statistical difference between the experiment and control groups was determined, indicating that Bright Blue had no effect on the neural cells and their repairing power. This rapid immunostaining technique offers a good approach for the identification and accurate coaptation of sensory fasciculi in peripheral nerve repair.

     

神经近端吻合术防治外伤性截指后顽固性残端神经痛

目的　报告神经近端吻合术在外伤性截指后顽固性残端神经痛的防治。方法　采用神经近端吻合术 ,预防及治疗外伤性截指术后顽固性残端神经痛 6 4例。术后随访 5 2例。其中 ,急诊手术 35例 ,1指 2 1例 ,2指 8例 ,3指 6例。晚期手术 17例 ,1指 9例 ,2指 4例 ,3指 2例 ,4指 2例。结果　术后得到随访者无 1例发生顽固性残端神经痛。结论　神经近端吻合术是防治外伤性截指术后顽固性残端神经痛的有效方法     

周围神经端侧吻合再生纤维来源的实验研究

目的:探索周围神经端-侧吻合后再生纤维(侧芽)的来源。方法:将Wistar大鼠左侧腓总神经切断后端-侧吻合到外膜开窗的胫神经干上,术后8周对胫神经和腓总神经分别用快兰和核黄作神经干注射标记,取脊髓背根神经节(DRG)荧光显微镜下观察被标记细胞;取吻合口附近的胫、腓神经段经锇酸染色后在光镜下观察腓总神经内轴突与胫神经轴突的关系。结果:实验组背根节内未发现双标细胞。光镜下,可清晰地见到左侧腓总神经内的再生纤维来自于胫神经近侧端。结论:周围神经端-侧吻合腓总神经内的再生纤维来自于胫神经的侧芽缺乏依据,因而无法排除来自腓总神经近端生长锥和受损胫神经纤维生长锥的可能。     

臂丛神经根性撕脱伤动物模型的实验研究

目的：为臂丛神经实验外科提供更加科学准确的动物模型。方法：60只SD大鼠，随即分为3组。A组：切断肌皮神经主干。B组：切断颈6及加入其中的颈7分支后将其近端撕脱。C组：切断颈5、6、7后将其近残端尽量拉出剪断。左例为实验侧，右例为正常侧，进行自身对照。术后定期检测电生理学、组织学及肌肉功能等指标。结果：A组和C组术后实验侧与正常对照侧相比各项指标均有显著性差异；B组术后早期肱二头肌虽有轻微改变，但晚期实验侧与正常对照侧相比各项指标无显著性差异。结论：切断大鼠肌皮神经主干，或者切断颈6的同时将颈5和颈7切断，然后近端撕脱，可以准确模拟臂丛神经根性撕脱伤。     

大鼠膈神经异化支配迷走神经的Y管实验研究

目的探讨大鼠膈神经经Y形管长入迷走神经的异化再生特征及神经生长因子（nerve growth factor,NGF）对其再生的影响。方法SD大鼠20只,颈部建立膈神经一迷走神经“Y形再生室”异化神经模型（硅胶Y管近端接膈神经近端,远端接迷走神经和膈神经远端）,随机分成2组,A组（对照组）10只,术后腹腔注射生理盐水,B组（NGF组）10只,术后腹腔注射NGF,连续2周。术后12周电刺激检测大鼠的心脏功能,对再生神经进行组织学观察。结果电刺激再生异化神经后。B组大鼠动脉血压及心率的下降幅度均大于A组,差异有统计学意义（P〈0．05）;B组再生异化神经和膈神经有髓纤维数目、通过率、髓鞘厚度和轴突直径均大于A组,差异有统计学意义（P〈0．05）;同组两侧比较,再生有髓神经数目和通过率为异化神经侧多,髓鞘厚度和轴突直径为膈神经侧大,差异有统计学意义（P〈0．05）。结论异化神经具有成熟髓鞘结构及靶器官支配功能,在性质上属于躯体运动神经;NGF能够促进异化神经生长并提高其支配效能;再生神经无明显解剖部位趋化性,长入远端神经干粗大侧（迷走神经）有数量优势,长入躯体运动神经侧（膈神经）有质量优势。     

An experimental study of using “tendon bridge” or “tendon tunnel” as a conduit for nerve regeneration

In our experiments, a “tendon bridge” or a “tendon tunnel” was used as a conduit to repair the defect of a peripheral nerve in rabbits. A gap of about 1 cm was made on the deep peroneal nerve in each animal. In group 1, the tendon of anterior tibial muscle was isolated near the severed nerve, and a segment of half cross section of tendon was removed in ladder-shaped reversed fashion. The proximal and distal nerve stump were sutured respectively to the tendon by 11/0 atraumatic sutures under surgical microscope. In group 2, the nerve stumps were sutured to the tendon of anterior tibial muscle without cutting the tendon, but the tendon was dissected free and turned to form a tunnel wrapping the gap in between the nerve stumps. The results showed that a gap of 1 cm of the deep peroneal nerve might be repaired by both methods, but the results of using “tendon tunnel” in group 2 were much better than those of using “tendon bridge” in group 1.     

神经再生中靶器官的诱向作用

本研究以细胞培养的方法证明了在周围神经再生过程中,远侧端即轴突衍生的靶器官,很强地诱导着神经轴突的定向生长。进而采用了自然凝胶电泳系统,分析了与神经诱向再生相关的活性因子,其结果提示在远侧端组织中,出现的18KD和89KD因子具有很强的诱向性作用,诱导轴突在再生的过程中准确到达靶器官;并很强地提示雪旺氏细胞在神经再生过程中扮演着重要角色。     

周围神经43kD蛋白在损伤坐骨神经中的表达

目的：探索SD大鼠损伤坐骨神经组织中43kD蛋白的表达。方法：采用周围神经43kD蛋白单克隆抗体,以Western Blot方法检测大鼠损伤坐骨神经中相应蛋白的表达情况。结果：正常大鼠坐骨神经组织与大鼠损伤后2周的坐骨神经远侧端组织,在43kD处均有免疫反应阳性条带,在大鼠损伤后2周的坐骨神经远侧端组织中阳性反应产物着色较深。结论：大鼠坐骨神经组织中有43kD蛋白的表达,而损伤神经远侧端43kD蛋白表达增强。     

Experimental studies of specific neuro-chemotaxis in peripheral nerve regeneration in the rat

In this paper are reported the results of a systematic study of chemotactic effect and other features of the regeneration of severed peripheral nerve in rat. The results revealed that (1) the proximal stumps of tibial and peroneal nerves tended to grow to their original distal stumps, (2) motor branch and sensory branch also showed preferential growth to the homojunctional patterns of nerve grafts, (3) using muscle and skin as targets, these not only attracted nerve growth, out also selectively attracted motor branch and sensory branch. Our results indicate that during rat peripheral nerve regeneration there exists neurotropism as evidenced by tissue specificity, topographic specificity, functional pattern specificity and target specificity. Based on these findings, the possible mechanism of neurochemotropism is discussed. Distance between the stumps, selective use of experimental chamber material and the clinical significance of this neurotropic research are also discussed.     

应用端侧吻合方法治疗失神经肌肉萎缩的实验研究

目的 探讨应用端侧吻合方法防治失神经肌肉萎缩的作用,为临床治疗提供实验依据.方法 选用SD雌性大鼠作为研究对象,共32只,分为四组,每组8只.A组:将右侧腓总神经切断后,于相邻胫神经干上行外膜“开窗”,将腓总神经远侧断端以端侧吻合的方式吻合于胫神经干上.B组:将一神经移植段的两端分别与正常胫神经干和切断腓总神经远端行外膜“开窗”端侧吻合.C组:将右侧腓总神经切断后,两断端均结扎并翻转缝合于临近肌肉上.D组:正常对照组:大鼠右侧腓总神经不做任何处理.术后12周行神经肌肉形态学检查,胫前肌肌纤维横截面积,肌湿重及神经电生理检测,评价端侧吻合方法对失神经肌肉萎缩的防治作用.结果 ①神经肌肉形态学检查:A,B,D三组均可见再生的轴突和髓鞘,并且见正常肌肉形态结构,C组神经远断端纤维化并且肌纤维萎缩变细.②胫前肌肌纤维横截面积:A,B两组间差异无显著性(P＞0.05),C组与A,B,D三组比较,差异有显著意义(P＜0.05)③胫前肌肌湿重:A,B两组较D组稍轻,统计学上无显著差异(P＞0.05),C组与A,B,D三组比较,差异有显著意义(P＜0.01)④神经传导速度比较:C组测不出运动神经传导速度,A,B两组运动神经传导速度慢于D组,A,B两组间运动神经传导速度比较,无统计学意义(P＞0.05),A,B两组与D组比较,差异有统计学意义(P＜0.01).结论 端侧吻合法为防治肌肉失神经萎缩的有效方法,但其再生神经对肌肉支配功能不足以代替原支配神经.     

Mechanism of the inhibition of neuroma formation by autogenous nerve replantation in situ after amputation in rabbits

Two experimental patterns of autogenous nerve replantation in situ and centrocentralnerve suture were performed on the rabbit median and ulnar nerves.The results of light andtransmission electron microscopy as well as immunohistochemistry showed that autogenous nervereplantation in situ could inhibit the formation of neuroma.The mechanism might be that twoproximal stump axons could touch within the interpolated graft segment.Simple end-to-endnerve suture failed to inhibit neuroma development because of a lack of proper environment forthe touching between two proximal stump axons.     

兔自体神经原位移植限制创伤性神经瘤形成机理

本实验采用家兔近端正中神经和尺神经原位移植法研究预防创伤性神经瘤形成之机理。经光镜,透射电镜及免疫组化标记观察表明:(1)自体神经原位移植法能有效地抑制创伤性神经瘤形成。两神经再生的神经纤维在移植段中排列整齐,来自两神经的轴索末端髓鞘对接是抑制创伤性神经瘤形成的关键;(2)移植段中由于雪旺氏细胞增生所形成的雪旺氏鞘对近端神经轴索的再生有趋化和引导作用;(3)单纯吻合法不能抑制创伤性神经瘤的发生。     

睫状神经营养因子对面神经再生影响的实验研究

目的：以新西兰白兔面神经损伤为动物模型,研究睫状神经营养因子（CNTF）对损伤后面神经再生的作用,为临床上选择高效的神经营养因子提供实验依据。方法：选用28只新西兰白兔,将其双侧面神经上颊支离断后套入硅胶管,吻合神经断端,并将硅胶管两站与神经外膜吻合后,向套管内注射CNTF液（实验组）以及生理盐水（对照组）。于术后1,2,4,8周采用电生理、光镜、透射电镜等手段来检测。结果：术后2周、4周,实验组不论是神经传导速度还是组织学形态均明显优于对照组。8周以后,实验组和对照组无明显差别。结论：局部应用外源性CNTF短期内可明显促进损伤面神经的再生,有很大的临床应用价值。     

血小板源性生长因子B在损伤周围神经再生中的作用

目的：观察血小板源性生长因子B（PDGF-B）在损伤大鼠坐骨神经中的表达,旨在阐明其在末梢神经再生中的作用。 方法：分别构建大鼠坐骨神经切割伤和碾压伤模型,应用免疫组织化学S-P法检测PDGF-B的表达。结果：两类损伤模型中,损伤近端神经和再生轴突PDGF-B的表达均增强。两类损伤模型均可见损伤神经远端的雪旺氏细胞PDGF-B表达量明显增加,在碾压伤模型中,随着轴突-雪旺氏细胞关系的恢复,PDGF-B表达水平下降,并最终恢复正常;在切割伤模型中,二者关系不能恢复,PDGF-B减至极低水平。 结论：坐骨神经损伤后PDGF-B的表达增强在末梢神经再生中发挥重要作用。