周围神经虚拟三维重建中神经束功能及形态定位的组织化学染色方法研究

目的探索周围神经虚拟三维重建中不同性质神经纤维与功能束的组织学水平的形态学定位方法。方法取自愿捐献新鲜成人尸体右侧正中神经作为样本进行连续冰冻切片,取形态完整标本切片30个,首先采用单纯Karnovsky-Roots法对神经切片染色(A组,n=30),再依次行甲苯胺蓝染色(B组,n=28)以及丽春红2R染色(C组,n=21)。每种染色完成后即于光学显微镜下分区显微摄影(×100)并拼接成全景图像,比较不同染色方法下全景图像的纹理特征、乙酰胆碱酯酶活性部位数量及平均灰度;并对比计算机自动获取神经束轮廓结果,以确立获取理想图像的染色方案。结果 A、B、C组乙酰胆碱酯酶活性位点数量分别为(21.63±4.06)×102、(20.64±3.51)×102、(20.54±5.71)×102个;平均灰度分别为(1.41±0.06)×102、(1.10±0.05)×102、(1.14±0.07)×102。各组间乙酰胆碱酯酶活性位点数量差异无统计学意义(F=0.64,P=0.54);与A组比较,B、C组平均灰度均较低,差异有统计学意义(P<0.001)。A组仅乙酰胆碱酯酶活性部位着色;B组髓鞘显示不满意;C组可同时显示神经纤维轴突及髓鞘染色,神经束和不同性质神经纤维纹理特征最显著,神经束边界轮廓最清晰,计算机处理时假阳性容易去除,图像分割最精确。结论 Karnovsky-Roots-甲苯胺蓝-丽春红2R三重复染方法不影响神经纤维乙酰胆碱酯酶阳性位点的表达,图像纹理清晰,较符合周围神经虚拟三维重建时在组织学水平分辨及获取神经束功能状态二维图像的相关要求,有望解决周围神经组织形态学表达方式这一技术难题。     

神经内松解术对周围神经卡压作用的实验研究

评估神经内松解术对周围神经卡压的效果。方法：将大鼠坐骨神经卡压１２周后，分别采用单纯减压、神经内松解、神经内松解加地塞米松等３种方法手术。于术后第４、８、１２周作神经电生理检测，组织形态学观察。结果显示：内松解术组的电生理指标，神经纤维的直径、数目，髓鞘的厚度及纤维结缔组织减少的程度均优于同期的单纯减压     

分子免疫染色法鉴别人体周围神经感觉纤维的实验研究及临床应用

用分子免疫染色法在术中鉴别人体周围神经感觉纤维。方法：在动物实验已成功的基础上，分离提取新鲜人体脊神经元特异蛋白，将其作为抗原制备了鼠抗人感觉神经元特异蛋白本克隆抗体。用食用亮蓝标记单克隆抗体后，直接对人体周围神经于断面进行染色的实验研究；并为临床６例周围神经损伤患者进行了术中染色，整个染色过程仅需３０分钟。结果：感觉神经纤维呈清晰蓝色，从而获得了满意结果。结论：该法特异性强，显色快，应用范围广，是鉴别周围神经功能束的一种特异、可靠、准确的方法     

远端神经变性对神经再生放大规律的影响

目的 探讨应用正常供体神经修复预变性受体神经时,再生神经纤维放大的效果.方法 将雄性SD大鼠12只,随机分为变性组、对照组,所有动物均实行手术.变性组应用部分正常腓总神经修复变性8周胫神经远端;对照组应用部分正常腓总神经修复即刻损伤后胫神经远端.术后3个月进行神经电生理测量、肌力测量、组织学观察、神经锇酸染色及有髓神经纤维计数,并计算两组神经再生放大率.结果 变性组和对照组神经传导速度分别为(16.992±3.737)m/s和(23.092±2.788)m/s;肌肉强直收缩力恢复率为(39.642±5.865)%和(71.098±6.778)%;再生有髓神经纤维数分别为1718.2±282.0和3340.0±506.5;神经再生放大率1.581±0.329和3.098± 0.642;以上指标变性组均低于对照组(P＜0.05).结论 应用正常神经修复8周变性神经远端时,神经纤维再生放大率降低,说明损伤远端神经较长时间变性,接受再生神经纤维长入的能力降低.     

FK506缓释膜片促进外周神经再生的实验研究

目的观察神经断伤吻合口局部植入FK506缓释膜片对神经再生的影响.方法取Wistar大鼠60只,随机平均分为A、B、C、D 4组.切断大鼠右侧坐骨神经后间断吻合,将免疫抑制剂FK506溶入聚乳酸(PLA)后制成15 mm×8 mm×1 mm大小的膜片,植入神经吻合口周围作为生物降解缓释制剂,可稳定、持续释放FK506约3周.A组为对照组,膜片内不含FK506;B、C、D组设计释放率分别为1 mg@kg-1@d-1、2 mg@kg-1@d-1和5 mg@k-1@d-1.术后第3、6、10周观察神经吻合口质量、测定坐骨神经功能指数以及电生理学和组织学检查的定性、定量分析.结果B、C组测定结果明显优于A组,C组优于B组.D组神经连接及再生不良.结论适量FK506神经吻合口植入,缓释应用对周围神经再生的速度和质量均有明显的促进作用.     

纤维蛋白胶粘合修复周围神经损伤的实验研究

目的 比较纤维蛋白胶粘合法与神经外膜缝合法修复周围神经损伤的效果。方法 将雄性Wistar大鼠20只随机分为两组，即粘合组和缝合组，每组10只。以大鼠左侧坐骨神经为修复神经模型进行实验。术后8周，各组取8只大鼠进行电生理、组织学检查，每组其余2只取标本做透射电镜观察。结果 术后8周粘合组神经纤维较缝合组平直，粘合组与缝合组比较，运动神经潜伏期延迟比、吻合口神经纤维通过比、有髓纤维截面积恢复比、肌湿重恢复比差别具有显著性意义(t值分别为2．32、2．43、2．39、2．36，P值分别为0．032、0．029、0．030、0．031)。结论 应用纤维蛋白胶粘合修复周围神经损伤是一种较为有效的神经接合方法。     

用组织扩张器延长损伤后的周围神经远段的实验观察

目的 观察用谐和顺延长损伤后谱性的周转神经远段作为修复神经缺损的实验结果。方法 以容积为１２ｍｌ外形近似长方体的组织扩张器,３周内缓慢延长１５只ＳＤ习一侧钳夹并结扎损伤的坐骨」神经远段,然后切去延长部分,使两种神经断端在无张力下缝合,术后１４周,进行电生理测定和组织学观察,并与对侧神经缺损后在张力下缝和比较。结果 和组织扩张器延长的损伤后坐骨神经远段,可延长３６．５％。再生神经纤维可通过变性后延长     

Rerouting peripheral nerves for spinal cord lesions

To provide control of paralyzed limb muscles following spinal cord lesions, peripheral nerves containing motor axons from motoneurons located above a spinal cord lesion could potentially be rerouted to nerves containing motor axons located below the spinal cord lesion. To test this hypothesis in rats, the distal end of a cut tibial nerve, innervated by the L4-6 spinal level, was anastomosed or rerouted to the central end of the cut femoral nerve, innervated by the L3-4 spinal level. Appropriate controls were used. Recovery of lower hind limb motor function was followed at regular intervals, measuring the twitch tension of toe flexion (innervated by the tibial nerve) induced by transcutaneous stimulation of nerve rootlets exiting the spinal cord. After 4-6 months, 54% of motor function returned in the experimental group. Retrograde transport of horseradish peroxidase from the gastrocnemius muscles to spinal motoneuron cell bodies confirmed that the innervation of this group was at a higher level. Furthermore, after an L4 spinal transection, twitch tension responses to spinal cord outlet stimulation remained only in the experimental group. Therefore, a peripheral nerve containing motor axons from above the lesion was rerouted to a distal peripheral nerve to control muscles that would have otherwise been denervated.     

周围神经移植修复脊髓损伤的实验研究

目的：探讨自体坐骨神经移植修复脊髓损伤的可行性。方法：将58只雌性Wistar大鼠分为二组，实验组：采用显微外科技术，将50只大鼠于T13水平切除左半侧脊髓10mm，再取右侧坐骨神经10mm移植到脊髓缺损处，近端接脊髓，远端接马尾，分别于术后2、4、6、8、12、22周在光镜和电镜下观察移植处坐骨神经、吻合口远端马尾神经、左后肢坐骨神经再生情况，并用摄像机记录患肢功能恢复情况。对照组：8只大鼠，于13水平切除左半侧脊髓10mm，不移植坐骨神经，观察脊髓缺损远端马尾神经和左右肢坐骨神经再生情况。结果：对照组坐骨神经的轴突及髓鞘部分崩解，密度降低，无再生轴突形成。实验组术后4周电镜下偶见移植处坐骨神经髓鞘及轴突形成，术后8周光镜及电镜下可见较多细的有髓神经纤维，22周时接近正常；同时观察到左后肢坐骨神经再生；大鼠后肢功能部分恢复，肌力达3级。结论：大鼠脊髓损伤后有再生能力，周围神经移植修复脊髓损伤是可行的。     

Repairing peripheral nerve defects with tissue engineered artificial nerves in rats

Objective： To observe the effect of tissue engineered nerves in repairing peripheral nerve defects （ about 1. 5 cm in length） in rats to provide data for clinical application.
Methods： Glycerinated sciatic nerves （2 cm in length） from 10 Sprague Dawiey （SD） rats （aged 4 months） were used to prepare homologous dermal acellular matrix. Other 10 neonate SD rats （aged 5-7 days） were killed by neck dislocation. After removing the epineurium, the separated sciatic nerve tracts were cut into small pieces, then digested by 2.5 g/L trypsin and 625 U/nd collagenase and cultured in Dulbecco＇ s modified Eagle＇ s medium （DMEM） for 3 weeks. After proliferation, the Schwann cells （SCs） were identified and prepared for use. And other 40 female adult SD rats （ weighing 200 g and aged 3 months） with sciatic nerve defects of 1.5 cm in length were randomly divided into four groups： the defects of 10 rats bridged with proliferated SCs and homologous dermal acellular matrix （the tissue engineered nerve group, Group A）, 10 rats with no SCs but homologous dermal acellular matrix with internal scaffolds （Group B ）, 10 with autologous nerves （Group C ）, and the other 10 with nothing （the blank control group, Group D）. The general status of the rats was observed, the wet weight of triceps muscle of calf was monitored, and the histological observation of the regenerated nerves were made at 12 weeks after operation.
Results： The wounds of all 40 rats healed after operation and no death was found. No foot ulceration was found in Groups A, B and C, but 7 rats suffered from foot ulceration in Group D. The triceps muscles of calf were depauperated in the experimental sides in all the groups compared with the uninjured sides, which was much more obvious in Group D. The wet weight of triceps muscle of calf and nerve electrophysiologic monitoring showed no statistical difference between Group A and Group C, but statistical difference was found between Groups A and B and Groups B and     

Immunology of peripheral nerves and response to trauma

Research in limb reconstruction using peripheral nerve tissue has been hampered by tissue rejection. In order to provide more information on the immunology of peripheral nerves, 35 specimens (26 from seven cadavers and nine from surgical biopsies) were analyzed for the presence of human leukocyte antigens (HLA), which are a key component of major histocompatibility complex class II (MHC Class II) antigens. MHC Class II antigens were noted in all human nerve specimens, and the percentage of areas of positive staining by immunohistochemistry was significantly higher (3.82% +/- 1.01%) than that of negative controls (.05% +/- 0.02%) (p less than 0.01). The negative controls consisted of serial sections that were stained in the same manner except that the primary antibody was deleted. There was no significant difference in the presence of MHC Class II antigens between sensory and mixed motor nerves or among the different HLA groups (HLA-DR, HLA-DP, HLA-DQ). A mouse model was used to evaluate the effect of trauma on the presence of MHC Class II antigens. Three weeks after transsection of the sciatic nerve there was a statistically significant increase (p less than 0.05) in the presence of MHC Class II antigens in the distal portion of the nerve (4.49% +/- 0.78%) as compared with samples from animals that had had exposure of the nerve without transsection (1.44% +/- 0.21%) or in nerves from animals that had had no surgery (1.02% +/- 0.21%). The presence of MHC Class II antigens could require more complex cross-matching for tissue transplantation. Nerve grafts that have undergone Wallerian degeneration may actually be more prone to rejection than are other peripheral nerve tissue transplants.     

甘露醇和当归对周围神经再灌注损伤的保护作用

目的　研究甘露醇和当归对周围神经再灌注损伤的保护作用。方法　观察神经电生理,神经内钙,自由基含量和组织结构的变化。　结果　缺血６ 小时应用甘露醇, 当归,再灌注５ 小时后潜伏期由（３７５６ ±０２０６） ｍ ｓ 减至（ ２２１６ ±００９６） ｍ ｓ 和（ ３４７２ ±０１６６ ） ｍ ｓ , 传导速度由（１２４１８ ±１０１１）ｍ ／ｓ 增至（ ２１３２０ ±０６７６ ） ｍ ／ｓ 和（ １７１２０ ±１０３５ ） ｍ ／ｓ , 波幅 由（ １２６２ ±０２５６ ） ｍ Ｖ 增 至（７１０８ ±０４３ ９） ｍ ｖ 和（２６０４ ±０２９ ７） ｍ Ｖ , 神 经内钙 由（ ７７７９４ ±８ ２６８ ） μｇ／ｇ 减 至（ ４２１３８ ±３４８９） μｇ／ｇ 和（４００９４ ±２０８２） μｇ／ ｇ 。缺血６ 小时应用当归,甘露醇再灌３ ０ 分钟, 自由基由（３７５４５ ±４４７５） Ｙｍ ａｘ／μｇ 减至（２８００９ ±２１７６） Ｙｍ ａｘ／ μｇ 和（ １１１３６ ±１８ ０６） Ｙｍ ａｘ／ μｇ 。组织形态学的变化也说明损伤程度得到明显抑制。　结论　甘露醇和当归均可用于防治周围神经的再灌注损伤, 甘露醇的作用优于当归     

Evaluating the effect of polytetrafluoroethylene and extractum cepae-heparin-allantoin gel in peripheral nerve injuries in a rat model

BACKGROUND:Peripheral nerves can be injured by congenital, mechanical, thermal or chemical causes. Peripheral nerve injuries are increasing in frequency, particularly in countries that are becoming more industrialized. Nerve and extremity injuries result in work loss and high treatment costs, and can lead to separation of patients from their social environment. Failure of nerve repair causes muscle functional losses, sensory losses and painful neuropathies.OBJECTIVES:To compare the effects of condensed polytetrafluoroethylene (cPTFE) and cPTFE-extractum cepae-heparin-allantoin (cPTFE-EHA) gel compound on nerve and functional recovery, and the prevention of adhesion and scar tissue formation after total peripheral nerve injury repaired by primary suture in a rat model.RESULTS:cPTFE alone and cPTFE-EHA gel was found to provide better functional recovery and nerve regeneration compared with primary repair only. In the macroscopic evaluation, the cPTFE-EHA gel was found to have no negative effect on wound healing and, despite increasing extra-neural scar tissue and adhesions, it had no negative effect on nerve function; in addition, it facilitated functional recovery.CONCLUSIONS:Compared with the cPTFE application alone, the application of perineural cPTFE-EHA gel during peripheral nerve surgery appeared to provide better functional recovery without causing any significant changes in epineural and extraneural scar tissue formation.     

Malignant epithelioid peripheral nerve sheath tumor arising in a benign schwannoma

The case of a patient with malignant degeneration of a solitary abdominal schwannoma and endobronchial metastasis is presented. The patient presented clinically with dyspnea referable to her lung mass, anorexia, and night sweats. The lung mass, initially diagnosed as a large-cell undifferentiated carcinoma, was later found to be histologically identical to the malignant portion of the abdominal tumor. The light microscopic, electron microscopic, and immunoperoxidase staining characteristics of the tumor are reported, and previous reports in the literature are reviewed.     

周围神经再生时乙酰胆碱受体免疫电镜与肌功能观察

目的 了解周围神经再生时Ｎ－乙酰胆碱受体（Ｎ－ＡｃｈＲ）改变与肌功能恢复关系。方法 选用家犬测定喉返神经再生时肌功能及肌电位,采用透射免疫电镜结合图像分析岂肉接头（ＮＭＪ）及其Ｎ－ＡｃｈＲ分布和含量改变。结果 术后３周,突前膜结构基本消失而后膜结构尚完整,Ｎ－ＡｃｈＲ含是 健侧增多,术后５周,再生神经长入ＮＭＪ,并可记录到肌电位,但Ｎ－ＡｃｈＲ含量处于最低状,肌收缩微弱;术后１２周,Ｎ－ＡｃｈＲ增     

丙酸睾酮对萃取神经修复大鼠坐骨神经缺损的促进作用

目的 观察丙酸睾酮对甘油萃取神经修复大鼠坐骨神经缺损的促进作用.方法 取SD大鼠60只,手术造成左侧坐骨神经1 cm缺损.每组20只.A组:甘油萃取的坐骨神经修复左侧缺损并注射丙酸睾酮(TP)50 g/L,0.1 ml,2次/周;B组仅以甘油萃取的坐骨神经修复缺损;C组以同种异体神经修复.术后16周,比较各组运动神经传导速度恢复率(RRMNCV)和腓肠肌复合动作电位波幅恢复率(RRCMAP);再生有髓神经纤维计数恢复率(RRMFP)及患侧小腿三头肌湿重/健侧小腿三头肌湿重率(ITSWW/UTSWW).结果 术后16周,A、B、c三组RRMNCV分别为(59.06±10.33)%、(45.80±9.05)%、(23.45±4.60)%;RRCMAP分别为(76.64±5.01)%、(64.55±4.20)%、(20.44 ±3.82)%;RRMFP分别为(79.11±2.55)%、(63.67±4.97)%、(29.76±5.68)%;ITSWW/UTSWW分别为(70.37±9.54)%、(56.77±6.08)%、(33.75±7.89)%,三组比较差异有统计学意义(P<0.05).结果 说明A组修复效果最好,B组次之,C最差.结论 丙酸睾酮对甘油萃取神经修复大鼠坐骨神经缺损具有促进作用.     

甲基泼尼松龙对周围神经修复的影响

目的 探讨甲基泼尼松龙对周围神经再生的促进作用及剂量选择。方法 28只成年SD雌性大，取股外侧切口，钳夹造成双侧坐骨神经损伤。创伤后1h起分别肌注不同剂量的甲基泼尼松龙或对照溶液，并间隔3h维持肌注，持续24h。按肌注剂量的不同随机分为大剂量组（首剂150mg/kg）、中剂量组（首剂30mg/kg）、小剂量组（首剂15mg/kg）和对照组（生理盐水与0．9％苯甲醛1：1混合液）。肌注后观察各组大鼠一般状态，记录术前及术后4周大鼠足印，测定坐骨神经功能指数（SFI0；术后4周显露双侧坐骨神经测定神经传导速度（NCV）；对取得的数据进行统计学分析。同时行神经横切片组织学观察。结果 中、小剂量组SFI均值优于大剂量组和对照组，但差异无显著性意义；中、小剂量组神经诱发电位引出率（100％）明显高于对照组（57．1％）（P＜0．01）；小剂量组NCV值优于其他三组，而且差异具有显著性意义（P＜0．05）；大剂量组大鼠均出现明显神经精神症状，有1只因感染死亡，3只4侧出现局部感染，其他三组未见明显异常。中、小剂量组神经横切片轴索再生情况优于大剂量组及对照组。结论 甲基泼尼松龙的全身应用可有效促进周围神经损伤后再生，但剂量选择不应过大，以减少感染及精神症状等副作用。     

Regeneration of peripheral nerves following Nd-YAG laser and scalpel transection—Experimental observations

Regeneration of 70 rat sciatic nerves was investigated following transection by a 1.06 μm neodymium-doped yttrium aluminium garnet (Nd-YAG) laser and by a conventional scalpel. Twenty-three amputation stumps after each technique were reexposed at different intervals between 3 and 240 days. In an second series 12 nerve trunks each were taken out following initial epineural suture repair after survival periods between 30 and 90 days. Light and electron microscopic findings were evaluated. Amputation neuromas could be observed in 16% of laser and 83% of scalpel cases. Laser neuromas were small and without adherence to surrounding tissue structures. Following end to end coaptation the repair site was crossed by sprouting axons in 92% of laser cases and in all nerves after scalpel division. Laser transection and suture repair resulted in optimal coaptation without fusiform thickening by proliferation of epineural fibroblastic tissue. Scar reactions with compression of the microfascicular pattern was present in 8% of laser and 75% of scalpel cases.     

The functional recovery of peripheral nerves following defined acute crush injuries.

This study evaluates the effect of crushing load on functional recovery of the sciatic nerve. Male Sprague-Dawley rats were divided into five groups: sham operation, resected sciatic nerve, and 100 g (13 mm Hg/mm2), 500 g (50 mm Hg/mm2), and 15,000 g (1,000 mm Hg/mm2) of sciatic crush load (groups 1-5). In groups 3-5, a 5-mm segment of sciatic nerve was crushed for 10 min using a specially designed crushing device. Motor functional recovery was assessed from hind-limb walking tracks by calculating a sciatic functional index. There was no detectable functional deficit in the group receiving sham operations, while the resected sciatic nerve group exhibited complete dysfunction for the full duration of the experiment. All groups subjected to crush exhibited an initial deficit that gradually recovered to normal by day 14 (100-g crush), day 39 (500-g crush), and day 53 (15,000-g crush). Histological changes were also related to the initial crushing load and the length of the recovery period. Results indicate that the crushing device described is able to administer an adjustable, defined crush injury to the rat sciatic nerve, and that the functional deficit resulting from such an injury can be easily monitored with a sciatic functional index. The rate of recovery of crushed nerves was directly related to the initial load. All crushed nerves recovered in this experiment, even after the application of a 15,000-g load for 10 min.     

体神经-内脏神经吻合后神经纤维再生过程的光镜电镜观察

目的：观察大鼠体神经-内脏神经吻合后神经纤维的再生。方法：人工建立体神经-内脏神经反射弧大鼠模型，用电镜配合光镜观察12只大鼠术后1、4、8、24周神经变性与再生。结果：术后8、24周大体观察见神经吻合口位置稍许膨大，光镜观察发现术后8周吻合口位置可见新生的轴突，电镜观察见术后1周吻合口及其远近段神经纤维发生Waller变性，术后4周吻合口部位有再生的有髓和无髓纤维，术后8周新生的髓鞘进一步增厚，板层结构清晰可见，术后24周，髓鞘成熟，轴浆富含微管、微丝、线粒体。结论：体神经运动纤维能够再生长入并替代内脏神经节前纤维；再生的神经纤维具备基本正常的周围神经超微结构。