不同病理类型的臂丛神经损伤后变性神经中S-100蛋白的变化

目的探讨不同病理类型的臂丛神经损伤后变性神经中Ｓ－１００蛋白的变化规律，为临床进行术中检测与判定提供理论依据，以进一步指导临床，提高臂丛神经损伤的诊治效果。方法建立ＳＤ大鼠不同病理类型的臂丛神经损伤，用免疫组织化学方法分别在损伤后１、２、３和６个月检测变性的远端神经中Ｓ－１００蛋白的分布和含量变化。结果Ｓ－１００蛋白主要沿轴突四周分布，节后组损伤１个月后Ｓ－１００蛋白呈阴性反应，而节前组术后６个月仍有阳性轴突。结论Ｓ－１００蛋白主要分布于雪旺细胞，节前与节后这两种不同病理类型的臂丛神经损伤中神经变性的程度不同，节前损伤远端神经在术后仍能持续保持Ｓ－１００蛋白阳性。     

不同病理类型臂丛损伤后变性神经中胶原的变化

目的研究不同病理类型的臂丛损伤后变性神经中胶原蛋白的变化规律。方法建立ＳＤ大鼠不同病理类型的臂丛损伤模型，用免疫组织化学和电镜方法，分别检测变性的远端神经中Ｉ型蛋白含量的变化及观察胶原纤维的超微结构。结果胶原含量在节前组中术后６个月组略上升，节后组术后１个月组即开始上升，３个月以上上升显著。节后组胶原含量多于节前组（Ｐ＜０．０１）。结论节前损伤远端神经中胶原增生较节后损伤轻且速度缓慢     

臂丛神经节前损伤的大鼠动物模型

目的：介绍大鼠臂丛神经节前损伤动物模型的设计及实验结果。方法：选用ＳＤ大鼠，通过扩大椎间孔撕脱脊神经节，造成臂丛神经节前损伤模型（实验组），用组织病理学方法观察神经节内神经元细胞的变化，并用免疫组化方法，检测神经中Ｓ－１００蛋白阳性轴突数及灰度。结果：节前损伤模型可见撕脱的神经节，术后 １个月仍能见到神经节及神经元细胞； Ｓ－ １００蛋白阳性轴突在术后 １个月仍有表达；而节后损伤（对照组）到术后１个月则未见Ｓ－１００蛋白阳性轴突。结论：为臂丛神经损伤的深入研究提供一个具有科学性及合理性的实验模型。     

自体,异体雪旺细胞及其培养液植入羊膜基底膜修复神经缺损的？…

目的 比较自体雪旺细胞、异体ＳＣ、ＳＣ培养液对周围神经损伤后的再生作用。方法：取４０只ＳＤ大鼠,将其坐骨神经切除１ｃｍ造成神经缺损。用羊膜基底膜桥接后,分成４组,在间膜基底膜管中分别植入：（１）自体ＳＣ;（２）异体ＳＣ;（３）ＳＣ培养液;（４）生理盐水,为对照组,术后１个月和３个月,各组分别进行神经－肌电图和组织学观察。结果 术后３个月,自体ＳＣ组,ＳＣ培养液组的运动神经传导速度比术后１个月增快１     

自体、异体雪旺细胞及其培养液植入羊膜基底膜修复神经缺损的比较研究

目的比较自体雪旺细胞（Ｓｃｈｗａｎｎｃｅｌ,ＳＣ）、异体ＳＣ、ＳＣ培养液对周围神经损伤后的再生作用。方法取４０只ＳＤ大鼠,将其坐骨神经切除１ｃｍ造成神经缺损。用羊膜基底膜桥接后,分成４组,在羊膜基底膜管中分别植入：（１）自体ＳＣ;（２）异体ＳＣ;（３）ＳＣ培养液;（４）生理盐水,为对照组。术后１个月和３个月,各组分别进行神经－肌电图和组织学观察。结果术后３个月,自体ＳＣ组、ＳＣ培养液组的运动神经传导速度比术后１个月增快１２．１％、１１．２％;运动诱发电位波幅比术后１个月增高１４．９％、４．７％;而异体ＳＣ组和对照组则呈下降趋势。组织学发现：术后１个月４组均有神经轴突长过桥接段。术后３个月自体ＳＣ组、ＳＣ培养液组神经纤维数量增多、轴突直径增粗、髓鞘形成较好。结论自体ＳＣ和ＳＣ培养液对周围神经的再生有很好的促进作用。机体对异体ＳＣ会产生排斥反应,暂不宜采用。     

大鼠阴茎海绵体nNOS神经纤维损伤后再生可能性的研究

目的 探讨海绵体神经损伤对阴茎海绵体ｎＮＯＳ神经纤维的影响。方法 采用免疫组织化学技术ＳＰ法测定大鼠海绵体神经损伤后阴茎勃起组织ｎＮＯＳ神经纤维数量。结果 双侧损伤组术后３周,阴茎海绵体ｎＮＯＳ神经纤维显著减少,６个月后仍很少;单侧损伤组,术后３周损伤侧ｎＮＯＳ神经纤维的改变类似于双侧损伤组,６个月后损伤侧ｎＮＯＳ神经纤维明显增多,接近于对侧水平。结论 阴茎海绵体ｎＮＯＳ神经纤维在单侧海绵体神经损     

慢传输型便秘结肠神经丝蛋白和S—100蛋白的表达及意义

目的：探索结肠慢传输型便秘的发病机制。方法：采用免疫组织化学方法研究３３例慢性传输型便秘的病人（ＳＴＣ组）和２５例非便秘性结肠（对照组）升、横、降及乙状结肠肌间神经丛内神经丝蛋白和Ｓ－１００蛋白的表达。并利用图像分析系统作定量分析。结果：ＳＴＣ组结肠ＨＥ染色显示肌间神经丛未见明显异常，免疫组织化学染色示，对照组各段肠壁肌间神经丛内神经丝蛋白和Ｓ－１００蛋白的含量较恒定（Ｐ〉０．０５）。ＳＴＣ组结肠各段与对照比较。神经丝蛋白的含量无明显减少（Ｐ〉０．０５），但出现堆积聚集现象，平均光密度明显高于对照组（Ｐ〈０．０１）；Ｓ－１００蛋白的含量及平均光密度明显高于对照组（Ｐ〈０．０１）。且随着病程的延长而增加，二者呈直线相关（Ｐ〈０．０２）。结论：ＳＴＣ结肠肌间神经丛的病理改变是全结肠性改变。神经丝蛋白的堆积聚集和神经     

慢传输型便秘结肠肌间神经丛形态学改变及其临床意义

目的：探讨慢传输型便秘（ＳＴＣ）结肠传输缓慢的神经病理学基础。方法：对１４例ＳＴＣ患者的乙状结肠全层组织进行常规ＨＥ染色、肌间神经丛Ｈｏｌｍｅｓ银浸染色以及Ｓ－１００蛋白免疫组化染色等研究。并与１１例非梗阻性直肠癌患者的正常乙状结肠标本作对照。结果：１４例ＳＴＣ患者乙状结肠肌间神经丛嗜银性反应较对照组均有不同程度的降低，神经纤维排列紊乱并有缠结现象。肌间神经丛内Ｓ－１００蛋白的免疫反应性在ＳＴＣ组     

脊柱侧凸矫正术中的神经损伤并发症

目的：分析脊柱侧凸矫正术中神经损伤的机理及防治措施。方法：５４例脊柱侧凸患者，男１８例，女３６例。Ｃ－Ｄ法矫正３３例，平均年龄１５．５岁，Ｃｏｂｂ角平均６６°；ＴＳＲＨ法矫正２１例，平均年龄１４岁，Ｃｏｂｂ角平均６８°。结果：Ｃ－Ｄ组矫正后Ｃｏｂｂ角减少至２６°，ＴＳＲＨ组减小至２８°。２例出现神经损伤（３．７％）。其中１例经唤醒试验发现神经损伤立即取出Ｃ－Ｄ棒，术后双下肢瘫痪。卧石膏床３个月，给予辅助性治疗，１年后基本恢复；另１例术中诱发电位显示波幅降低，但唤醒试验正常，术后右大腿有一过性感觉障碍，１周后恢复。结论：神经损伤应分为严重损伤和一过性损伤，前者应及时取出内固定器械，后者可严密观察。对畸形严重，尤其是双主弯者，不能一味追求矫正度数而忽视神经损伤的发生     

不同时间段的预变性神经段分子生物学变化

目的 了解预变性神经移植修复神经缺损较新鲜神经移植修复效果良好的机制,并研究预变性神经段分子生物学变化。方法 选用３６只ＳＤ大鼠,将大鼠双侧上肢尺神经在腋部切断,远断端埋入肱二头肌中。术后当天、３ｄ、１周、２周、３周、４周（１￣６组）取材,应用ｒｔ－ＰＣＲ,Ｓｏｕｔｈｅｒｎ Ｂｌｏｔ的方法检测不同时间段的预变性神经中ＢＤＮＦ和ＣＮＴＦ的变化。结果 预变性神经中ＢＤＮＦ和ＣＮＴＦ的变化有明确的特点。     

陈旧性神经损伤后雪旺细胞的变化实验研究

目的 观察大鼠坐骨神经损伤后雪旺细胞的变化。方法 切断成年SD大鼠右侧坐骨神经，形成10mm缺损。于术后1～12个月不同时间段取材。标本用p75受体（p75 neurotrophin receptor．p75NTR）、S-100免疫荧光染色，激光共聚焦显微镜观察，另部分标本进行超微结构观察。结果 神经损伤后1个月，损伤远端神经中p75NTR和S-100的表达最多，在损伤后6个月时p75NTR降至正常水平，S-100则在损伤后9个月时消失。电镜下，雪旺细胞出现在神经内膜管内，神经内膜管下及内膜管周围有大量胶原原纤维增生。结论 大鼠坐骨神经损伤后，雪旺细胞中p75NTR的表达增加，但随损伤时间的延长呈进行性下降，伤后6个月时消失。     

后根轴突再生重建感觉传入通路的实验研究

目的 将后根分别与脊神经节中枢突支(后根)和周围突支(脊神经)吻合,为通过后根再生重建感觉传入通路奠定组织学基础.方法 SD大鼠74只随机分为三组,左侧为正常对照侧,右侧为手术侧.A组将L4后根近断端L6后根远断端吻合;B组将L4脊神经出椎间孔2 mm处切断,近断端与L6后根远断端吻合;C组手术操作同B组,但不吻合作为假手术组.术后3个月以逆行神经示踪、脊髓后根入口(dorsal root entry zone,DREZ)降钙素基因相关肽(calcitonin gene-related peptide,CGRP)阳性轴突分布、脊神经节和脊髓后角神经元存活率、光镜和透射电镜评价轴突再生效果.结果 逆行神经示踪证实B组再生轴突轴浆运输功能恢复良好.A组和B组均可见轴突再生通过吻合口,神经纤维数量和髓鞘厚度与正常对照侧比较差异无统计学意义;但A组轴突再生停止于DREZ,脊髓后角CGRP阳性纤维分布密度低于B组和正常对照侧,差异有统计学意义.三组L6脊神经节神经元存活率与正常对照侧比较差异无统计学意义;A组和C组L4脊髓后角神经元存活率低于B组和正常对照侧,差异有统计学意义.结论 后根与脊神经吻合后不影响脊髓后角神经元存活,神经纤维再生、轴浆运输能通过DREZ进入脊髓,是一种理想的修复感觉传导通路的方式.

Abstract：

Objective To provide the morphological evidence for sensory pathway by the repair of dorsal root (DR) in which received transected and anastomosed with central processes (DR) and peripheral processes (spinal nerve) respectively. Methods 74 SD rats were divided into three groups randomly. Group A: the distal ending of L6 DR was sutured to the proximal ending of the L4 DR in right side. Group B: the postganglionic spinal nerve of L4 was cut at a point 2 mm to intervertebral foramen and its proximal stump was reconnected to the distal ending of L6 DR in right side. Group C: under the same operative manipulation on group B, but no anastomosis was performed to serve as the sham operation group. The left side was kept intact and served as a normal control in all groups. The repairing results of DR axonal regeneration were evaluated at 3 months after operation by retrograde tracing, the density of positive Calcitonin gene-related peptide (CGRP) axon in dorsal root entry zone (DREZ), survival rate of neurons in DR ganglia and posterior horn, light and transmission electron microscope. Results HRP retrograde tracing demonstrated axonal axoplasmic transport of regenerative nerve recovered well in group B. Regenerative fibers were found. There was no difference between group A or B and normal control with regard to the number of myelinated axons and the thickness of myelin. Regenerating axon stop at the DREZ in group A and the density of positive CGRP were lower than those in group B and normal control. Survival rate of neurons in L6 DR ganglia had no difference among all groups. However, in posterior horn, a significant increase in death was seen in group A and sham group. Conclusion The ability of axonal regeneration was effective and with no disservice to neurons in posterior horn following anastomosis of postganglionic spinal nerve and dorsal root, by which axoplasmic transport across the DREZ into the spinal cord.     

Markers of nerve tissue injury in the cerebrospinal fluid in patients with lumbar disc herniation and sciatica

STUDY DESIGN: The light subunit of neurofilament protein, S-100 protein, neuron-specific enolase, and glial fibrillary acidic protein were determined in the cerebrospinal fluid in patients with lumbar disc herniation and in control patients. OBJECTIVES: To determine whether nerve root injury caused by disc herniation increases the levels of nerve and glial cell injury markers in the cerebrospinal fluid. SUMMARY OF BACKGROUND DATA: Markers of nerve tissue injury can be analyzed in the cerebrospinal fluid, allowing characterization of the cell types involved and the degree of disease in patients with neurologic disorders. METHODS: Cerebrospinal fluid samples were obtained by preoperative lumbar puncture in patients who underwent surgery for lumbar disc herniation and in patients who underwent lower extremity surgery (control group), neurofilament protein (light subunit) and glial fibrillary acidic protein were analyzed by enzyme-linked immunosorbent assay and S-100 protein and neuron-specific enolase by radioimmunoassay and luminescence immunoassay, respectively. In the disc herniation group the concentrations of the four markers were evaluated regarding possible correlation to patient history, computed tomographic findings, and clinical findings. RESULTS: Cerebrospinal fluid concentration of neurofilament protein (light subunit) and S-100 were increased in the disc herniation group compared with that in control subjects (1158 +/- 383 ng/L vs. 152 +/- 14 ng/L, P < 0.01; 1963 +/- 231 ng/L vs. 1003 +/- 152 ng/L, P < 0.05, respectively). No statistical differences in neuron-specific enolase and glial fibrillary acidic protein concentrations were observed between the groups. Disc herniation patients with fewer than 3 months' duration of subjective symptoms had higher neurofilament protein levels than did patients with longer duration. None of the markers was related to preoperative clinical or computed tomographic findings. Patients with persistent neurologic findings at follow-up 2-3 months after surgery had higher levels of neurofilament protein before surgery compared with-those without sequelae. CONCLUSIONS: Patients with disc herniation and sciatica have increased concentrations of neurofilament protein and S-100 in the cerebrospinal fluid, which indicates damage of axons and Schwann cells in the affected nerve root.     

End-to-side nerve repair: review of the literature and clinical indications

End-to-side (ETS) nerve repair, in which the distal stump of a transected nerve is coapted to the side of an uninjured donor nerve, has been suggested as a technique for repair of peripheral nerve injuries where the proximal nerve stump is unavailable or a significant nerve gap exists. Full review of the ETS literature suggests that sensory recovery after ETS repair results in some, but not robust, regeneration. Sensory axons will sprout without deliberate injury. However, motor axons only regenerate after deliberate nerve injury. Experimental and clinical experience with ETS neurorrhaphy has rendered mixed results. Continued research into ETS nerve repair is warranted. ETS techniques should not yet replace safer and more reliable techniques of nerve repair except when some, but not good, sensory recovery is appropriate and a deliberate injury to the donor motor nerve is made.     

Effect of a modified Nd:YAG laser technique on neuroma formation: An experimental study in rat sciatic nerve.

BACKGROUND AND OBJECTIVES: Traumatic transection of a peripheral nerve is inherently associated with the development of neuroma at the end of the proximal stump, often leading to therapy-resistant pain. This study was designed to evaluate whether the neodymium:yttrium aluminum garnet (Nd:YAG) laser could prevent neuroma formation after neurectomy. STUDY DESIGN/MATERIALS AND METHODS: The sciatic nerves of 14 rats were diffuse coagulated by defocused Nd:YAG laser (12 W power), and subsequently transected with additional focused laser energy. The control group consisted of contralateral nerves transected by microscissors. The nerves were reexposed at different time intervals up to 9 weeks after surgery, and evaluation consisted of macroscopy, and light and transmission electron microscopy. RESULTS: True neuroma formation could not be observed after laser transection, and only five nerves formed a neuromatous bulb, with minimal adhesions to surrounding tissue. Microscissor transection resulted in widespread amputation neuromas, consisting of regenerating axons and connective tissue, and nervous tissue regenerating into surrounding tissue. Laser-transected nerves showed degenerative changes of the axons and myelin, while proliferation of Schwann cells could not be observed. No outgrowth of axons could be observed outside the coagulated proximal stump. An epi/perineurial layer was present, covering the nerve stumps. Microscissor-transected nerves showed proliferation of fibroblasts and Schwann cells, forming minifascicles, and vigorous outgrowth of axons into the tissue and even into the distal nerve stump. CONCLUSIONS: Within the limitations of this study it is concluded that the formation of amputation neuromas is suppressed by Nd:YAG laser application by thermal coagulation of the nerve and suppression of Schwann-cell proliferation.     

Comparison of the neurotropic effects of motor and sensory Schwann cells during regeneration of peripheral nerves.

We examined the inductive ability of motor and sensory Schwann cells on regeneration of motor and sensory axons using a silastic Y chamber, and Lewis rats L5 ventral root (motor) and saphenous nerve (sensory). We developed four experimental models: motor-motor nerve group-proximal motor stump with distal fresh and frozen/thawed motor nerve segments (n = 7); sensory-sensory nerve group-proximal sensory stump with distal fresh and frozen/thawed sensory nerve segments (n = 7); motor-sensory nerve group-proximal motor stump with distal fresh and frozen/thawed sensory segments (n = 8); and sensory-motor nerve group-proximal sensory stump with distal fresh and frozen/thawed motor segments (n = 8). The gap was set at 4 mm. Six weeks postoperatively we compared the number of regenerated myelinated axons in the two distal channels, and found that sensory Schwann cells have a strong inductive ability for regeneration of both sensory and motor axons. Motor Schwann cells have weak inductive ability for regeneration of motor axons and no inductive ability for regeneration of sensory axons.     

The influence of motor axon misdirection on muscle contraction force in early nerve repair in a rat sciatic nerve model.

We examined the influence of misdirection of regenerating motor axons toward the distal sensory Schwann tubes on the muscle contraction force in early nerve repair using a rat sciatic nerve model. At 0, 1, 2, 4 and 8 weeks after severing the tibial, peroneal and sural nerves, the proximal stump of the tibial nerve was anastomosed with the distal stumps of both the peroneal and sural nerves using tubulisation (n=10 in each of five groups). We intentionally used the distal stump of the sural nerve (a sensory nerve) to induce regeneration in motor axons from the proximal tibial nerve stump toward the distal sensory nerve stump. Twenty-four weeks after nerve repair, isometric contraction force and wet weight of the anterior tibial muscle were measured, and the numbers of regenerated myelinated axons (motor and sensory) in the distal sural and peroneal nerves were counted. The rates of sural nerve regeneration were significantly higher at weeks 0 and 1 than at the later repair times. However, muscle contraction force and muscle wet weight did not differ significantly between groups in which nerves were repaired within four weeks of severance. These results indicate that peripheral nerve repair within four weeks of severance does not influence the muscle contraction force of single muscle despite the misdirection of motor axons toward the distal sensory Schwann tubes.     

Serum S-100B protein in severe head injury.

OBJECTIVE: Despite the significant recent progress in cerebral monitoring, it is still difficult to quantify the extent of primary brain injury and ongoing secondary damage after head injury. The objective of our study was to investigate S-100B protein as a serum marker of brain damage after severe head injury. METHODS: Eighty-four patients with severe head injury (Glasgow Coma Scale score < or =8) were included in this prospective study. Venous blood samples for S-100B protein were obtained as soon as possible after admission and every 24 hours thereafter, for a maximum of 10 consecutive days. Serum levels of S-100B protein were compared with outcome after 6 months, clinical variables, and the category of the Marshall classification of initial computed tomographic findings. RESULTS: Patients who died had significantly higher serum S-100B values compared with those who survived (median, 2.7 microg/L versus 0.54 microg/L; P < 0.0001, Mann-Whitney U test). Nineteen (58%) of 33 patients who died had peak S-100B values of 2 microg/L or higher, compared with 4 (8%) of the 51 surviving patients (P < 0.0005, Fisher's exact test). There was also a strong correlation between S-100B values and computed tomographic findings. Logistic regression analysis in a model with age, Glasgow Coma Scale score, intracranial pressure, and computed tomographic findings revealed S-100B as an independent predictor of outcome. Persistent elevation of S-100B levels for 2 to 6 days, even in patients with favorable outcome, may reflect ongoing secondary damage after severe head injury. CONCLUSION: S-100B may be a promising serum marker for assessing the extent of primary injury and the time course of secondary damage after severe head injury.