神经肌腱吻合预防神经瘤形成的实验研究

目的：为预防神经瘤形成，治疗神经瘤性残端痛提供一种新的有效的方法。方法：利用ＳＤ大白鼠３０只进行动物实验，在显微镜下，神经残端与肌腱端端吻合。结果：通过光学显微镜及大体观察，用神经与肌腱吻合，吻合口没有神经瘤形成。结论：神经肌腱吻合可以使再生的神经纤维沿着腱纤维间隙生长，能防止神经瘤的形成，达到治疗神经瘤性残端痛的目的，建议在临床上应用。     

神经肌腱对吻法预防截指术后神经瘤性疼痛

目的探讨神经肌腱对吻法预防截指术后神经瘤性疼痛疗效。方法临床200例急诊人院手外伤需截指患者，按手术先后随机分成实验组（常规截指术中采用神经肌腱对吻法处理手指神经残端）和对照组（常规截指术未作处理），每组100例。结果术后200例中失访14例，186例随访6个月～2年，按残端指神经瘤性疼痛发生率，实验组明显低于对照组。结论神经肌腱缝合法处理手指神经末梢是一种有效的预防神经瘤性疼痛的方法。     

自体神经桥接陈旧性周围神经缺损后雪旺细胞的组织学观察

目的:探索长时间失神经损伤后的雪旺细胞的促神经再生功能。方法:切除成年雌性SD大鼠左侧坐骨神经4 mm,分别饲养3、6个月后,修剪近、远侧神经断端制成不同持续时间的大鼠坐骨神经10 mm陈旧性缺损模型。实验组切取对侧正常坐骨神经桥接缺损,对照组缺损模型制备完成后不予任何修复。各组动物术后再饲养3个月取材,标本进行神经三色染色、免疫荧光染色,电镜等组织学方法观察。结果:各组损伤近端神经结构无明显差异,实验组桥接物段可观察到粗细不等的有髓神经纤维。电镜下,实验组远侧断端皆可观察到髓鞘形成良好的有髓神经纤维和存活的雪旺细胞。结论:长时间失神经损伤的雪旺细胞仍能在一定时间内存活并维持其促神经再生功能。     

单环刺缢及同种异体神经制成人工神经修复神经缺损

目的:研究去细胞的单环刺缢体壁和同种异体神经所构成的人工组织神经的组织相容性及其修复神经缺损的效果。方法:取大鼠40只随机分成实验组、自体神经组、硅胶管组、正常组。实验组将去细胞的单环刺缢体壁缝合成神经导管,其内充填去细胞的异体神经,修复大鼠10 mm坐骨神经缺损。术后4月,通过大体观察,组织形态学观察,了解该人工组织神经修复大鼠坐骨神经缺损的疗效。结果:该人工组织神经组织相容性良好,神经功能恢复效果正常组>自体神经组>实验组>硅胶管组,实验组疗效与自体神经组接近,明显优于硅胶管组。结论:将去细胞的单环刺缢体壁和同种异体神经制成人工组织神经修复周围神经缺损是可行的。     

海藻纤维膜片促进大鼠损伤坐骨神经的再生

背景：课题组和青岛大学高分子材料研究所合作研制的海藻纤维生物膜,具有优良的生物相容性,常被用作制备各种复合材料。 目的：观察海藻纤维膜片包绕覆盖神经断端吻合口对大鼠坐骨神经损伤后再生的影响。 方法：切断36只雄性Wistar大鼠右侧坐骨神经,随机分组：对照组行神经外膜端端吻合;实验组行神经外膜端端缝合,将海藻纤维膜片包绕并覆盖神经吻合口远近端各约0.5 cm,形成封闭再生室。术后观察海藻纤维膜片降解吸收规律及缝合处粘连情况,组织学切片行苏木精-伊红染色、锇酸染色、白细胞介素2及白细胞介素4免疫组织化学染色。 结果与结论：术后4-6周,实验组海藻纤维膜片逐渐被降解吸收,与周围组织粘连较少,炎性细胞浸润程度较轻,纤维组织增生较少。两组术后1,7,14 d的白细胞介素2及白细胞介素4含量比较差异无显著性意义。实验组术后6周再生神经纤维分布规则且大小较为均一,其神经纤维数量、轴突大小及髓鞘厚度等指标均显著优于对照组(P < 0.05)。表明海藻纤维膜片具有良好的生物降解性和组织相容性,其包绕覆盖坐骨神经形成的神经再生密闭室可促进大鼠损伤坐骨神经再生。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

同种异体神经复合体修复兔周围神经缺损

背景：应用种植许旺细胞的去细胞同种异体神经复合体修复周围神经缺损,探索其对神经再生及功能恢复有更好的促进作用,并且免疫原性非常小。 目的：用种植胎兔许旺细胞的去细胞同种异体神经复合体修复兔缺损的坐骨神经,观察移植神经周围免疫细胞的变化及功能恢复。 　 方法：48只新西兰白兔随机分成实验组和对照组。两组动物均切除一段坐骨神经,造成2.0 cm长的缺损,实验组用种植胎兔许旺细胞的同种异体神经复合体修复坐骨神经;对照组仅用去细胞同种异体神经修复。移植后1,4,8周光镜观察移植段坐骨神经周围肌肉组织中免疫细胞的浸润情况,计数每个高倍视野免疫细胞的数量。移植后4,8,16周大体观察兔的足部溃疡形成及愈合情况,大体观察神经愈合情况;肌电图检查桥接段坐骨神经的传导速度。 结果与结论：手术区局部均未出现明显的排斥反应,实验组足部溃疡愈合情况优于对照组。移植后1周移植段坐骨神经周围肌肉组织中有大量淋巴细胞及巨噬细胞浸润,实验组明显多于对照组(P < 0.05);移植后4周,浸润的免疫细胞两组均较1周后明显减少,实验组减少更明显。移植后8周,浸润的免疫细胞更加减少,但两组间比较差异无显著性意义(P > 0.05)。移植后4周时,两组均未见明显的神经传导,8,16周神经传导速度实验组均优于对照组(P < 0.05)。提示,种植许旺细胞的去细胞同种异体神经复合体免疫原性非常小,对神经再生及功能恢复有更好的促进作用。     

以化学去细胞法处理同种异体神经与周围静脉伴行修复面神经缺损

背景：有研究表明化学去细胞法处理的同种异体神经修复面神经缺损可以取得较好的修复效果。 目的：在化学去细胞法处理同种异体神经的基础上探索一种修复面神经缺损更有效的修复方式。  方法：将新西兰大白兔随机分为2组,实验组制备面神经颊支缺损动物模型,采用经化学去细胞的同种异体腓肠神经进行移植,且与伴行静脉行外膜缝合;对照组在同样位置的远近端分别切断面神经,但不破坏被切断神经与周围组织的正常解剖关系,再切断处行外膜缝合桥接。 结果与结论：修复后3个月两组兔均存活,面部表情基本对称,胡须活动正常,神经移植处未见明显瘢痕及神经瘤形成。电镜观察结果显示,实验组与对照组右侧面神经颊支传导速度,移植体远端吻合口附近5.0 mm段有髓神经纤维数量,靶肌肉运动终板计数均差异无显著性意义(P > 0.05)。结果证实,化学去细胞同种异体神经与周围静脉伴行修复家兔面神经缺损的方法可以达到与自体面神经原位移植相似的修复效果。     

静脉桥接法治疗及预防创伤性神经瘤

目的：评价静脉桥接法治疗及预防残端创伤性神经瘤的临床疗效。方法：采用静脉桥接法治疗及预防创伤性神经瘤14例。根据患者术后疼痛缓解程度、局部有无触痛、Tinel＇s征及患者的满意程度等对术后效果做出评价。结果：14例患者术后随访时间为3个月～5a，自觉疼痛症状均得到明显缓解，无自发性疼痛，局部无触痛。结论：静脉桥接法是治疗创伤性神经瘤的有效方法。     

吻合血管的尺神经转位治疗截瘫

目的：重建胸段脊髓横断致伤完全性截瘫患者的部分周围神经功能。方法：在４例截瘫病人将一侧的尺神经自腕部切断,以骨间前神经旋前方肌支吻接远端尺神经深支,取一桡神经浅支的分支与尺神经浅支吻接,自皮下隧分段游离尺神经至腋窝起始处,分离过程中保护尺侧上副血管。于皮下将尺神经及其尺侧上副血管引人侧胸壁隧道。显露胸背动,静脉,将尺侧上副血管与其行端端吻合。在臂部分离出阴部神经、股后皮神经和坐骨神经,用带筋膜蒂股     

聚乳酸和聚羟基乙酸共聚物复合膜修复坐骨神经损伤

背景：聚乳酸和聚羟基乙酸复合膜具有良好的生物相容性,稳定的机械强度,无毒副作用,可控的降解速率等特点。 目的：观察聚乳酸和聚羟基乙酸共聚物膜对大鼠坐骨神经损伤的修复作用。 方法：手术显露36只Wistar大鼠坐骨神经,随机分成3组：假手术组游离坐骨神经后不作任何处理,对照组切断坐骨神经后行神经断端直接吻合,实验组于神经吻合断端包裹聚乳酸和聚羟基乙酸共聚物膜。 结果与结论：①神经电生理学检测：术后4,6周神经传导速度及波幅比较,对照组<实验组<假手术组(P均<0.05)。②组织学检测：对照组神经与周围组织粘连严重,实验组吻合口光滑平整,聚乳酸和聚羟基乙酸共聚物膜明显降解吸收,与周围组织无粘连,对照组有髓神经数量较假手术组、实验组明显减少,且轴突再生率和再生轴突成熟度较低。③辣根过氧化物酶逆行示踪检测：对照组被辣根过氧化物酶标记的阳性有髓神经纤维数量较假手术组、实验组明显减少。表明聚乳酸和聚羟基乙酸共聚物膜能减轻神经术后粘连,促进神经再生。     

生长相关蛋白及其mRNA在大鼠损伤坐骨神经中的表达

为了研究大鼠周围神经损伤后远侧端组织中生长相关蛋白 (GAP-4 3 )及其 m RNA的表达 ,采用正常 SD大鼠坐骨神经横断损伤的模型 ,于术后不同时间取坐骨神经远侧端组织 ,以 Western Blot和 RT-PCR法检测 GAP-4 3及其 m RNA的表达。RT-PCR法检测结果显示 ,正常大鼠坐骨神经组织中 GAP-4 3 m RNA表达量较低 ,而损伤坐骨神经的远侧段 GAP-4 3 m RNA的表达量明显增加 ,于损伤第 2周时达高峰。用抗 GAP-4 3抗体进行 Western Blot检测 ,结果表明 ,正常坐骨神经 43 k Da处出现弱阳性反应的蛋白区带 ,而损伤后坐骨神经远侧段 43 k Da处阳性反应条带着色明显加深 ,损伤后第 3周时阳性反应着色最强。本研究结果提示 ,GAP-4 3及其 m RNA在大鼠损伤坐骨神经远侧段组织中表达增强 ,其 m RNA表达上调高峰在神经损伤后的第 2周 ;而其蛋白合成增加最为明显的时间是在神经损伤后的第 3周。     

生长相关蛋白在损伤坐骨神经中的表达

目的：研究大鼠周围神经损伤与再生过程中生长相关蛋白的表达。方法：取正常SD大鼠坐骨神经和离断损伤后不同时间近侧端和远侧端神经组织，采用Anti-GAP－43抗体以Western blot方法检测GAP－43的表达变化。结果：NC膜上43kDa位置出现阳性反应条带，在正常坐骨神经中反应微弱，损伤后明显加强，随损伤时间延长，神经近侧端的反应无明显改变，神经元侧端的反应逐渐加强，以第3周为最强，之后逐渐减弱，结论：GAP－43在正常坐骨神经中低表达，损伤后近侧端与远侧端表达均增加，远侧端的表达增加以第3周为高峰。     

p27kip1和Skp2在损伤后大鼠坐骨神经中的表达变化

为了探讨损伤后周围神经p27kip1和S期激酶相关蛋白2(Skp2)的定位表达和变化,本实验将成年SD大鼠随机分为正常对照组、夹伤组和切断组,运用Western blot结合免疫组织化学及免疫荧光双标,在大鼠坐骨神经损伤时,对p27kip1和Skp2表达的影响进行了研究。结果表明:(1)坐骨神经夹伤后,p27kip1蛋白表达先逐渐下降,后又逐渐上升;坐骨神经切断后,远侧段p27kip1蛋白表达持续下降,而近侧段p27kip1蛋白表达在切断后6h明显下降,后又逐渐升高至正常水平,而Skp2表达变化与之相反;(2)免疫组织化学染色结果显示,坐骨神经切断后1w,远侧段从断端到末端,p27kip1阳性信号逐渐增加,而Skp2阳性信号逐渐减弱;(3)免疫荧光双标显示,正常和损伤坐骨神经的雪旺氏细胞中都有p27kip1和Skp2表达。以上结果提示:周围神经损伤后影响雪旺氏细胞中p27kip1和Skp2的表达变化,为进一步研究它们在周围神经损伤和修复中的作用机制奠定基础。     

The Protective Effect of Vipera Raddei Venom on Peripheral Nerve Damage

Acute experiments were performed on spinal rats to study the protective actions of Vipera raddei venom after section of the sciatic nerve. Individual spike activity was recorded from interneurons and motoneurons in the lumbar segment of the spinal cord, induced by stimulation of the sciatic nerve and the extensor (gastrocnemius) and flexor (peroneus communis) nerves on the lesioned and symmetrical intact sides in controls and after daily injections of venom for four weeks. In animals not treated with Vipera raddei venom, the lesioned side lacked interneuron and motoneuron responses to stimulation of the extensor and flexor nerves of the distal stump, though these were present on stimulation of the contralateral side; responses were the inverse of this on the intact side, due to the failure of the proximal and distal stumps to fuse, as also demonstrated by atrophy of the distal stump of the sciatic nerve and the absence of movement activity in the lesioned limb. Treatment with Vipera raddei venom led to restoration, by four weeks, of interneuron and motoneuron responses on the lesioned side on stimulation of the ipsilateral nerves and on the intact side by stimulation of the contralateral nerves; this is the result of apparent fusion of the proximal and distal stumps of the lesioned nerve. Further evidence for this was hypertrophy of the distal stump and restoration of movement activity in the lesioned limb. These results show that Vipera raddei venom has potential for use in regenerating damaged peripheral nerves. __________ Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 90, No. 12, pp. 1441–1456, December, 2004.     

基于高频交流电刺激的神经传导阻断的动物实验研究

目的：面肌痉挛虽不危及生命，但可以对患者造成心理负担，严重影响患者正常的工作和生活。高频电刺激可以有效阻断运动神经的神经冲动，从而终止相应的神经功能。本研究通过利用动物在体神经进行实验的方法研究高频信号阻断神经传导的效应，为高频电刺激治疗面肌痉挛提供更多的信息。方法：采用Wistar大鼠在体坐骨神经进行实验，对神经施加高频阻断刺激信号，通过记录并观察神经所支配肌肉的肌电信号及其肌肉状态来确定阻断效果．研究高频交流电信号对周围神经传导阻断的作用。结果：当对神经给予高频阻断刺激信号时，神经出现传导阻断的现象，且阻断现象发生于某一频段，完全阻断则只是发生于其中更小的频率范围，而继续增大频率或幅值，则可能会出现部分阻断、强直甚至不阻断现象。结论：本动物实验研究为将来高频电刺激治疗面肌痉挛的临床应用的实现提供更多信息。     

大鼠坐骨神经损伤后诱导型一氧化氮合酶表达的变化

为了探讨大鼠坐骨神经损伤后再生过程中诱导型一氧化氮合酶(iNOS)表达的变化及意义,本研究采用大鼠坐骨神经切断缝合模型,分别于术后1、3、7、14、21及28d取吻合口远端的神经,采用免疫组织化学和实时荧光定量聚合酶链反应(RT-PCR)方法检测损伤神经远端iNOSmRNA及其蛋白的表达水平。结果显示:假手术对照组坐骨神经中未见明显的iNOS阳性产物,iNOSmRNA表达极低。实验组神经损伤后iNOSmRNA及其蛋白的表达水平均明显增高(P<0.01),iNOS阳性产物的吸光度(A)值在术后7d达高峰。iNOSmRNA表达在术后1、3、7d维持较高水平,此后则明显下降。上述结果说明大鼠坐骨神经损伤后神经纤维中iNOS的表达增加,iNOS可能在周围神经损伤后的再生过程中起着一定的作用。     

Spatiotemporal Expression of SSeCKS in Injured Rat Sciatic Nerve

SSeCKS (src suppressed C kinase substrate) functions in the control of cell signaling and cytoskeletal arrangement. It is expressed in brain and spinal cord, but little is known about its expression in peripheral nerves. In this study, in rats, real‐time polymerase chain reaction and Western blot analysis showed that expression of SSeCKS in crushed sciatic nerve reached its highest level 6 hr after crushing, whereas in a transection model, SSeCKS peaked at 2 days in the proximal stump and 12 hr in the distal stump. Immunohistochemical analysis demonstrated up‐regulation of SSeCKS protein surrounding the crush site and in the two stumps of the transected nerve. In addition, SSeCKS colocalized with growth‐associated protein 43 and with S100, which also changed with time after injury. These findings support the idea that SSeCKS participates in the adaptive response to peripheral nerve injury and may be associated with regeneration. Anat Rec, 291:527–537, 2008. © 2008 Wiley‐Liss, Inc.     

Peripheral Nerve Regeneration and Electrophysiological Recovery with CIP-Treated Allogeneic Acellular Nerves

Acellular nerve grafts are a desirable alternative to autografts, both because the source of acellular nerves is potentially unlimited and because they have the same matrix structure as natural nerves, which would facilitate axon growth from the defective nerve stump. Although some acellular nerves have been developed, most of them were studied in isogenic transplantation models and evaluated only by histological observation. In the present study, novel allogeneic acellular nerves prepared using the cold isostatic pressuring (CIP) method were developed and assessed as a potential substitute for autografts. The host immune response to acellular nerves and fresh nerves was analyzed using Lewis rats as donors and SD rats as recipients, which is the allogeneic transplantation model, by subcutaneous implantation for one month. In addition, sciatic nerve transplantation into a 10-mm nerve gap was carried out using the same model, and the axonal growth in acellular nerve transplantation was evaluated histologically and electrophysiologically, and compared with that of axons in the autograft transplant area. The subcutaneously implanted acellular nerves contained more macrophages and less vasculature than the allogeneic fresh nerves. In spite of these results of the subcutaneous implantation, Schwann cell infiltration in the graft transplanted into the sciatic nerve gap was observed after the short-term transplantation. The myogenic potential, which was measured as an index of electrophysiological function in acellular nerve transplantation, was also recovered in the long-term transplantation. Our results indicate that the acellular nerves developed herein have the potential to support nerve regeneration and might be useful as an alternative to autografts.     

Effect of nerve length and temperature on the induction of action potentials in denervated slow muscle fibres of the frog

A. Pyriformis and extensor longus digiti IV muscles of Rana temporaria were denervated by cutting the sciatic or peroneal nerve at various distances from the muscles. Slow fibres were identified by their membrane time constants, and examined for their ability to produce action potentials in response to intracellularly applied current pulses. B. The slow muscle fibres acquired the ability to generate action potentials several days after denervation. The duration of this latent period depended on the length of the peripheral nerve stump, and on the temperature at which the frogs were kept after the operation. C. At 18 degrees C the latent period increased by 0.36 days per mm of sciatic nerve stump. At 11.5 degrees C the corresponding value was 0.7 days/mm. The effect of length of the peroneal nerve was smaller than that of the sciatic nerve. D. It is suggested that the peripheral nerve stump serves as a reservoir of 'trophic' material which is transported towards the slow fibres at a rate of 2.8 mm/day (at 18 degrees C) and seems to block the formation of Na channels. The Q10 value of this transport system would be 2.7.