生殖股神经移植重建大鼠勃起神经通道

目的　探讨利用生殖股神经移植加神经生长强化介质修复损伤的阴茎海绵体神经,重建勃起神经通道的可行性。　方法　3月龄SD雄性大鼠54只,随机平均分成假手术对照组、双侧阴茎海绵体神经损伤组及神经移植加胰岛素样神经生长因子Ⅰ(IGF Ⅰ)组。术后1、3、6个月用阿朴吗啡试验评估各组动物勃起功能,并取阴茎干组织,采用NADPH d组化法了解nNOS阳性神经纤维的再生情况。　结果　术后1、3、6个月,假手术对照组均有正常阴茎勃起(勃起率100% ),神经切断组完全丧失勃起功能(勃起率0% );术后1个月神经移植组也丧失勃起功能(勃起率0% ),术后3、6个月勃起功能渐恢复(勃起率分别为50%和66. 7% ),两两比较差异有统计学意义(P<0. 05)。术后3、6个月,神经移植组nNOS 阳性神经纤维数显著增多,神经部分切断组的nNOS阳性神经纤维数和术后1个月相比无增多,两两比较差异有统计学意义(P<0. 002)。　结论　双侧阴茎海绵体神经损伤后,利用生殖股神经移植加IGF促进神经再生,是重建勃起神经通路,恢复大鼠勃起功能的一种有效方法。     

海绵体神经移植恢复大鼠勃起功能的研究

目的　应用腓肠神经移植替代损伤的双侧海绵体神经恢复大鼠的勃起功能。方法　将 48只雄性SD大鼠随机分为 3组 :神经移植组、神经损伤组及假手术组。 2、4个月后 ,海绵体神经电刺激检测大鼠勃起功能 ,免疫组织化学法检测海绵体内nNOS阳性神经纤维。结果　 2个月后神经移植组与神经损伤组大鼠对海绵体神经电刺激均无勃起反应 ,两组海绵体内nNOS阳性神经纤维数目差异无显著性 (P >0 .0 5 ) ;而 4个月后神经移植组大鼠勃起功能较神经损伤组差异有显著性 (P <0 .0 5 ) ,海绵体内nNOS阳性神经纤维数目也显著高于神经损伤组 (P <0 .0 5 ) ,与假手术组差异无显著性 (P >0 .0 5 )。结论　腓肠神经移植替代损伤的双侧海绵体神经可恢复大鼠的勃起功能。     

海绵体神经损伤所致ED大鼠模型建立

目的 :寻找大鼠海绵体神经并建立神经损伤所致ED大鼠模型。　方法 :对 2 0只大鼠进行解剖 ,在外科显微镜下找到海绵体神经并经电刺激试验证实。随后将 4 2只实验大鼠随机分为假手术对照组、单侧海绵体神经损伤组及双侧海绵体神经损伤组。术后 3周用阿朴吗啡试验来评估所建动物模型。　结果 :盆大神经节位于背侧前列腺后外侧叶表面 ,海绵体神经是最大的传出神经。诱发阴茎勃起的电刺激参数 :电压 5V、刺激频率 2 0Hz及刺激时间 5ms。术后 3周 ,阿朴吗啡均能诱发对照组大鼠阴茎勃起 ,30min内平均勃起 (2 5 7± 1 4 0 )次。实验组大鼠 ,无论单侧损伤还是双侧损伤 ,均丧失勃起功能 (0 0 0± 0 0 0 )。　结论 :大鼠较大的盆大神经节及海绵体神经易于辨认 ,电刺激反应明显 ,是建立海绵体神经损伤性ED模型的理想动物。无论是单侧海绵体神经损伤还是双侧海绵体损伤 ,损伤早期 ,大鼠均丧失勃起功能     

腓肠神经移植重建海绵体神经保留勃起功能的实验研究

目的 :研究腓肠神经移植替代损伤的双侧海绵体神经恢复大鼠的勃起功能。　方法 :4 8只雄性SD大鼠 (3～ 4月龄和 30 0～ 4 0 0g)随机分为神经移植组、神经损伤组及假手术组 ,每组 16只。 2、4个月后 ,海绵体神经电刺激检测大鼠阴茎勃起功能 ,阴茎内注射神经逆行示踪剂荧光金 5d后检测盆神经节内被标记的神经元细胞。　结果 :2个月后神经移植组与神经损伤组大鼠对海绵体神经电刺激均无勃起反应 ,两组盆神经节内荧光金标记的神经元细胞数目差异有显著性 (P <0 .0 5 ) ;而 4个月后神经移植组大鼠勃起功能较神经损伤组差异有显著性 (P <0 .0 5 ) ,盆神经节内荧光金标记的神经元细胞数目也显著高于神经损伤组 (P <0 .0 5 ) ,与假手术组差异无显著性 (P >0 .0 5 )。　结论 :腓肠神经移植替代损伤的双侧海绵体神经可恢复大鼠的勃起功能。     

神经套管术重建勃起功能的动物实验研究

目的　探讨利用神经套管术加神经生长强化介质修复阴茎海绵体神经损伤 ,重建勃起功能的可行性。　方法　 5 4只SD雄性大鼠随机分成假手术对照组、双侧阴茎海绵体神经损伤组及神经套管加胰岛素生长因子 (IGF)组 ,术后 1、3、6个月用阿朴吗啡试验评估所建动物模型的勃起功能 ,随后取阴茎干、脚组织 ,利用NADPH d染色法证实nNOS阳性神经纤维的再生情况。　结果　术后 1个月 ,阿朴吗啡诱导阴茎勃起试验中 ,神经部分切断组和神经套管IGF组间勃起率没有显著差异 ;术后 3、6个月 ,神经套管组勃起率分别为 4 2 %和 5 0 % ,神经部分切断组勃起率均值为 0 % (P <0 .0 5 )。术后 3、6个月 ,神经套管组nNOS阳性神经纤维数显著增多 ,神经部分切断组的nNOS阳性神经纤维数同术后 1个月相比无增多 (P <0 .0 0 2 ) ,两两比较差异有显著性意义。　结论　利用神经套管加IGF促进神经再生 ,是双侧阴茎海绵体神经损伤后 ,重建勃起功能的一种有效方法。     

骨髓间充质干细胞即刻与延时治疗修复阴茎海绵体神经损伤大鼠勃起功能的研究

目的:拟观察在双侧阴茎海绵体神经(CN)损伤的大鼠神经性勃起功能障碍(NED)模型中即刻和延时向阴茎海绵体注射骨髓间充质干细胞(BM-MSCs)对大鼠阴茎勃起功能的修复作用。方法:选取28只8周龄、体重200~250 g雄性SD大鼠随机分为4组:假手术组找到双侧海绵体神经后不做处理直接关腹并缝合皮肤;对照组、即刻治疗组和延时治疗组均通过钳夹的方式损伤双侧阴茎海绵体神经建立NED模型随后关腹缝合。假手术组和对照组向阴茎海绵体内注射对照剂,即刻治疗组注射BM-MSCs,延时治疗组术后两周注射BM-MSCs。术后12周采用电刺激CN记录阴茎海绵体内压(ICP),颈动脉穿刺测定平均动脉压(MAP),ICP/MAP作为勃起功能的评价指标来评估大鼠的勃起功能。在勃起功能检测后处死大鼠,取阴茎海绵体中段组织检测平滑肌、胶原和神经纤维。结果:在手术后12周,即刻治疗组和延时治疗组ICP和ICP/MAP均较对照组升高(P0.05)。即刻治疗组和延时治疗能提高大鼠海绵体组织内平滑肌与胶原的比例(P0.05),同时两组阴茎海绵体内平滑肌含量高于对照组(P0.05),阴茎海绵体背神经内神经微丝(NF)蛋白阳性神经纤维数目和nNOS的表达高于对照组(P0.05)。结论:阴茎海绵体内注射BM-MSCs能对双侧CN损伤大鼠的勃起功能恢复有促进作用。BM-MSCs治疗可以提高海绵体组织内平滑肌与胶原的比例,改善纤维化,并能提高海绵体组织中平滑肌含量、阴茎背神经的神经丝含量和nNOS的表达水平,术后延时治疗同样能取得一定的治疗效果。     

神经干细胞移植重建勃起功能的研究

目的 观察神经干细胞移植修复损伤的海绵体神经从而恢复勃起功能的可行性.方法 42只雄性SD大鼠(3～4个月龄和体质量300～400 g)随机分为假手术组、神经干细胞移植组和神经损伤组,每组14只.2、4个月后,海绵体神经电刺激检测大鼠阴茎勃起功能,免疫组织化学法检测海绵体内一氧化氮合酶(nNOS)阳性神经纤维.结果 2个月后3组大鼠对海绵体神经电刺激的勃起反应率分别为100%、0%和0%;3组海绵体内nNOS阳性神经纤维数目分别为98.5±9.2、22.5±3.5和25.7±5.1,后2组间差异无统计学意义(P>0.05).4个月后3组大鼠电刺激后勃起率分别为100%、57.14%和7.19%,后2组间差异有统计学意义(P<0.05);3组海绵体内nNOS阳性神经纤维数目分别为95.1±7.7、86.0±13.4、26.5±4.3,前2组间差异无统计学意义(P>0.05),后2组间差异有统计学意义(P<0.05).结论 神经干细胞移植是修复损伤的海绵体神经从而恢复勃起功能的一种有效方法.     

显微修复损伤的阴茎海绵体神经恢复大鼠勃起功能

目的　探讨利用显微外科技术修复外科损伤的阴茎海绵体神经 ,重建大鼠勃起通道的可行性。方法　 36只SD雄性大鼠随机平均分成 3组 ,即假手术对照组、双侧阴茎海绵体神经损伤组和显微修复组 ,术后 1、3个月后用阿朴吗啡 (APO)试验来评估所建动物模型 ,随后取阴茎干组织 ,利用NADPH d组化法证实nNOS阳性神经纤维的再生情况。结果　术后 1个月 ,神经部分切断组和显微修复组间差异无显著性 (P >0 .0 5 ) ;术后 3个月 ,APO所诱导的阴茎勃起试验中 ,显微修复组勃起率为 83 .3 % ;神经部分切断组勃起率一直为 0 % (P <0 .0 5 )。此外 ,神经套管组的nNOS阳性神经纤维数目在术后 3个月有显著增多 ,而神经部分切断组的nNOS阳性神经纤维数目同术后 1个月相比无增多 ,两两比较有差异有显著性 (P <0 .0 0 8)。结论　双侧阴茎海绵体损伤后 ,立即用显微外科技术吻合神经 ,是重建勃起通路 ,恢复勃起功能的一种有效方法。     

C7神经移位重建截瘫双下肢功能的实验研究及初步临床应用

目的 观察大鼠桡神经移位修复股神经，重建股四头肌功能的效果，并应用C7神经根移位．通过长段神经桥接修复股神经，重建截瘫患者双下肢的部分功能。方法 SD大鼠16只，随机分为两组。A组：将左侧桡神经切断．近端牵至锁骨下切口内．经吻合血管的坐骨神经进行桥接．一期修复同侧股神经；B组：行桡神经与桥接神经吻合．术后3个月再切断同侧股神经并与桥接神经吻合。观察指标为电生理、股四头肌湿重恢复率、肌纤维截面积恢复率、有髓神经纤维通过率及截面积恢复率。临床上应用C7神经根为动力神经源．将胫神经自小腿远端切断并向近端游离至臀部，应用带血管蒂的胫神经反转至颈部与C7神经吻合．待胫神经自颈部再生到臀部后，再将胫神经与股神经吻合来治疗2例截瘫患者，结果 B组股四头肌湿重．肌纤维截面积恢复率均忧于A组；两组有髓神经纤维通过率、截面积恢复率及肌肉复合动作电位波幅比较差异无显著性意义。1例随访41个月．左侧股四头肌肌力恢复至4^-级，右侧3级；另1例随访24个月．双侧股四头肌肌力恢复至2^-级。结论 周围神经具有强大的再生能力，C7移位分期修复股神经可重建截瘫患者下肢的部分感觉．运动功能。     

FK506对大鼠阴茎海绵体神经再生影响的研究

目的:探讨FK506对大鼠阴茎海绵体神经损伤后再生的影响,并讨论其作用的可能机制。　方法:54只 SD雄性大鼠随机分成3组,即假手术对照组(简称对照组,n=24)、单侧阴茎海绵体神经切断组(简称单切组,n= 24)、单侧阴茎海绵体神经切断+FK506组(简称FK506组,n=6)。术后1、3个月电刺激阴茎海绵体神经并连续监 测阴茎海绵体内压变化及阴茎勃起情况,同时取阴茎海绵体组织采用NADPH d法观察一氧化氮合酶(nNOS)阳性 神经纤维的再生情况。　结果:术后1个月单切组nNOS阳性神经纤维数量明显减少,与对照组相比差异有极显 著性(P<0.01),术后3个月在电刺激未切断侧阴茎海绵体神经时,FK506组比单切组产生更大的最大海绵体内压 (P<0.01),且单切组阴茎海绵体组织中nNOS阳性神经纤维比术后1个月无明显增加(P>0.05),而FK506组 nNOS阳性神经纤维显著增加(P<0.01)。　结论:FK506皮下注射能促进大鼠损伤的阴茎海绵体神经再生,促进 勃起功能恢复。     

Schwann cell seeded guidance tubes restore erectile function after ablation of cavernous nerves in rats

PURPOSE: Dissection of the cavernous nerves eliminates spontaneous erections. We evaluated the ability of Schwann cell seeded nerve guidance tubes to restore erections after bilateral cavernous nerve resection in rats. MATERIALS AND METHODS: Sections (5 mm) of the cavernous nerve were excised bilaterally, followed by immediate bilateral microsurgical reconstruction. In 10 animals per group (20 study nerves) reconstruction was performed by genitofemoral nerve interposition, interposition of silicone tubes or interposition of silicone tubes seeded with homologous Schwann cells. As the control 10 animals (20 study nerves) underwent sham operation (positive control) and bilateral nerve ablation (without reconstruction) was performed in a further 10 (negative control). Erectile function was evaluated 3 months postoperatively by relaparotomy, electrical nerve stimulation and intracavernous pressure recording. RESULTS: After 3 months neurostimulation resulted in an intact erectile response in 90% (18 of 20) of Schwann cell grafts, while treatment with autologous nerves (30% or 6 of 20) or tubes only (50% or 10 of 20) was less successful (p <0.01). Whereas untreated ablated rats showed no inducible erections (0% or 0 of 20), all sham operated animals had an intact erectile response (100% or 20 of 20). Maximum intracavernous pressure upon electrostimulation was significantly elevated using Schwann cell grafts compared to results in the other treatment groups (p <0.001). Morphological evaluation revealed advanced regeneration within Schwann cell grafts. CONCLUSIONS: Schwann cell seeded guidance tubes restore erectile function after the ablation of cavernous nerves in rats and they are superior to autologous nerve grafts.     

The effect of platelet-rich plasma on cavernous nerve regeneration in a rat model

The aim of this study was to investigate the effect of platelet-rich plasma (PRP) on cavernous nerve (CN) regeneration and functional status in a nerve-crush rat model. Twenty-four Sprague–Dawley male rats were randomly divided into three equal groups: eight had a sham operation, eight underwent bilateral nerve crushing with no further intervention and eight underwent bilateral nerve crushing with an immediate application of PRP on the site of injury. Erectile function was assessed by CN electrostimulation at 3 months and nerve regeneration was assessed by toluidine blue staining of CN and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining of penile tissue. Three months after surgery, in the group that underwent bilateral nerve crushing with no further intervention, the functional evaluation showed a lower mean maximal intracavernous pressure (ICP) and maximal ICP per mean arterial pressure (MAP) with CN stimulation than those in the sham group. In the group with an immediate application of PRP, the mean maximal ICP and maximal ICP/MAP were significantly higher than those in the injured control group. Histologically, the group with the application of PRP had more myelinated axons of CNs and more NADPH-diaphorase-positive nerve fibres than the injured control group but fewer than the sham group. These results show that the application of PRP to the site of CN-crush injury facilitates nerve regeneration and recovery of erectile function. Our research indicates that clinical application of PRP has potential repairing effect on CN and peripheral nerves.     

Bilateral cavernous nerve interposition grafting during radical retropubic prostatectomy: Memorial Sloan-Kettering Cancer Center experience

PURPOSE: Cavernous nerve graft is an option for men requiring bilateral cavernous nerve resection for cancer control during radical prostatectomy. We determined the success rate and identified determinants of success of bilateral cavernous nerve grafting following resection of the 2 nerves during radical prostatectomy in patients who were potent preoperatively. MATERIALS AND METHODS: We retrospectively reviewed the records of 44 consecutive patients who underwent bilateral nerve grafting from 1999 to 2004. Postoperative erectile function was defined as the achievement of erections satisfactory for intercourse with or without oral medication. We calculated cumulative erectile function recovery rates using Kaplan-Meier curves. The log rank test was used to compare variables affecting erectile function recovery with p <0.0083 considered significant after adjusting for the number of variables evaluated using the Bonferroni correction. RESULTS: The overall 5-year cumulative recovery of erectile function permitting penetration was 34% and the rate of consistent penetration was 11%. None of the analyzed variables were significantly associated with recovery of postoperative erectile function, including patient age (p = 0.3), incomplete bilateral cavernous nerve resection (p = 0.045), sural nerve grafts compared to genitofemoral or ilioinguinal nerves as donor sites (p = 0.067), post-radiation salvage radical prostatectomy (p = 0.15), neoadjuvant hormone therapy (p = 0.7) and comorbidities (p = 0.15) or medications (p = 0.4) affecting EF. CONCLUSIONS: Bilateral cavernous nerve grafts might be beneficial in select patients. A definitive answer awaits the performance of a multi-institutional, randomized, controlled trial.     

Bilateral nerve grafting during radical retropubic prostatectomy: extended follow-up

Objectives. To confirm the benefit of using an interposition sural nerve graft at the time of radical retropubic prostatectomy in an extended series of men with at least 1 year of follow-up. We previously reported the return of erectile function after resection of both cavernous nerves.Methods. Twenty-eight potent men with clinically localized prostate cancer underwent radical retropubic prostatectomy with deliberate wide bilateral neurovascular bundle resection and the placement of bilateral nerve grafts. Erectile dysfunction questionnaires and patient interviews were completed at 6-month intervals. A minimum of 12 months of follow-up (mean 23 ± 10 months) was obtained for 23 men (mean age 58 ± 6 years). A control group of 12 men who underwent bilateral nerve resections, but declined nerve graft placement, was also followed up.Results. Of the 23 men, 6 (26%) had spontaneous, medically unassisted erections sufficient for sexual intercourse with vaginal penetration. An additional 6 men (26%) described “40% to 60%” spontaneous erections (fullness, no rigidity, not able to penetrate). Ten men (43%) had intercourse with sildenafil. No demonstrable erections occurred before 5 months postoperatively. The greatest return of function thus far was observed at 18 months after surgery.Conclusions. This surgical technique continues to show promise as an advance in prostate cancer surgery. The results of this study demonstrated recovery of erectile function in men who underwent bilateral nerve graft placement during radical retropubic prostatectomy when both cavernous nerves were deliberately resected.     

Cavernous nerve regeneration using acellular nerve grafts

INTRODUCTION: The restoration of erectile function following complete transection of nerve tissue during surgery remains challenging. Recently, graft procedures using sural nerve grafts during radical prostatectomy have had favorable outcomes, and this has rekindled interest in the applications of neural repair in a urologic setting. Although nerve repair using autologous donor graft is the gold standard of treatment currently, donor nerve availability and the associated donor site morbidity remain a problem. In this study, we investigated whether an "off-the-shelf" acellular nerve graft would serve as a viable substitute. We examined the capacity of acellular nerve scaffolds to facilitate the regeneration of cavernous nerve in a rodent model. MATERIALS AND METHODS: Acellular nerve matrices, processed from donor rat corporal nerves, were interposed across nerve gaps. A total of 80 adult male Sprague-Dawley rats were divided into four groups. A 0.5-cm segment of cavernosal nerve was excised bilaterally in three of the four groups. In the first group, acellular nerve segments were inserted bilaterally at the defect site. The second group underwent autologous genitofemoral nerve grafts at the same site, and the third group had no repair. The fourth group underwent a sham procedure. Serial cavernosal nerve function assessment was performed using electromyography (EMG) at 1 and 3 months following initial surgery. Histological and immunocytochemical analyses were performed to identify the extent of nerve regeneration. RESULTS: Animals implanted with acellular nerve grafts demonstrated a significant recovery in erectile function when compared with the group that received no repair, both at 1 and 3 months. EMG of the acellular nerve grafts demonstrated adequate intracavernosal pressures by 3 months (87.6% of the normal non-injured nerves). Histologically, the retrieved regenerated nerve grafts demonstrated the presence of host cell infiltration within the nerve sheaths. Immunohistochemically, antibodies specific to axons and Schwann cells demonstrated an increase in nerve regeneration across the grafts over time. No organized nerve regeneration was observed when the cavernous nerve was not repaired. CONCLUSION: These findings show that the use of nerve guidance channel systems allow for accelerated and precise cavernosal nerve regeneration. Acellular nerve grafts represent a viable alternative to fresh autologous grafts in a rodent model of erectile dysfunction.     

猕猴去细胞同种异体神经体内移植的组织学观察

目的观察去细胞同种异体神经体内移植后组织形态学改变,寻找合适的人工神经支架材料。方法共使用成年猕猴9只,随机取其中的6只,培养自体Scs,应用已经制备好的去细胞神经为支架材料,体外构建组织工程化周围神经移植体修复猕猴尺神经40mm缺损。分实验组、空白组、正常对照组3组。术后5个月取材,观察大体形态、HE染色、免疫组织化学染色及组织形态学改变。结果3组移植物表面均有不同程度的血管化,HE染色均有细胞核和髓鞘的蓝染成份出现,NF免疫组织化学染色后,均有淡黄色神经微丝出现。实验组、正常对照组效果相似,它们均优于空白组、实验组,正常对照组再生神经纤维的数量差异均无统计学意义（P〉0．05）。但在实验组与空白组间的差异有统计学意义（P〈0．05）。结论去细胞同种异体神经具有良好的生物相容性和促进神经再生能力,是最具应用前景的神经修复材料之一。     

腹腔镜直肠癌根治术保留盆腔植物神经效果分析

目的：探讨在腹腔镜直肠癌根治手术中保留盆腔植物神经（PANP）对男性局部复发率、生存率和术后生活质量的影响。方法：不同时期的2组腹腔镜直肠癌根治手术病人,其中非保留盆腔植物神经组34例,保留组88例,回顾性分析比较2组患者的局部复发率、5年存活率及术后排尿功能和性功能。结果：两组均顺利完成手术。其中保留植物神经组82例获得随访,未保留组28例获得随访。局部复发率保留植物神经组为73％（6／82）,未保留组为7．1％（18／28）;5年存活率保留植物神经组为8412％（69／82）,未保留组为82．1％（23／28）,2组差异无统计学意义（P〉O．05）。排尿功能障碍保留植物神经组为28．1％（23／82）,未保留组为60．7％（17,28）;勃起功能障碍保留植物神经组为24．4％（20,82）,未保留组为67．9％（19／28）;射精功能障碍保留植物神经组24．4％（20／82）,未保留组71,4％,（20／28）,两组比差异均有统计学意义（PcO．05）。结论：腹腔镜直肠癌根治术中保留植物神经功能,对局部复发率和5年存活率无明显影响,但可明显提高患者的生存质量。     

端侧吻合中供体神经的损伤处理促进神经再生作用的研究

目的：研究端侧吻合中供体神经对于受体神经再生作用。方法：选用雄性SD大鼠20只,随机分为2组,每组10只,每组后肢共20侧。A组：左侧腓总神经的切断后,同时行在左侧胫神经待吻合口处先开窗后再横向切开一半,后将腓总神经远端以端侧吻合法45°缝合于胫神经开窗处（已横向切开一半处）。B组：除不作供体胫神经的横向切开一半的处理外,其余与A组处理相同。术后2个月行组织学检查,胫神经及腓总神经传导速度检查,腓总神经和胫神经电生理检查,胫前肌肌湿重量,电镜观察再生神经纤维数量。结果：两组肌湿重量无统计学意义（P〉0.05）,腓总神经的传导速度差异（P〈0.05）,腓总神经有髓神经纤维计数的差异（P〈0.01）,A组胫神经传导速度明显小于B组。结论：说明我们对供体神经进行横向切开处理对于神经再生是有必要的,但是对于胫神经的横向切开一半的处理对于胫神经产生了影响,在促进神经再生的同时需要重新对供体神经横向切开的范围进行更深的研究。