培养雪旺细胞NGF、CNTF和GDNF共表达和共定位实验研究

目的 探讨将雪旺细胞和背根节神经元共同培养后去除背根节神经元后雪旺细胞神经生长因子（nerve growth factor，NGF）、睫状神经营养因子（ciliang neurotrophic factor，CNTF）和胶质细胞源性神经营养因子（glial cell line-derived neurotrophic factot，GDNF）共表达变化特点及其细胞共定位。方法 运用多彩色荧光原位杂交和计算机图像处理技术实现雪旺细胞NGF、CNTF和GDNFmRNA共表达变化的定量分析和细胞共定位，并通过免疫组织化学方法验证有关结果。结果 NGF、CNTF和GDNFmRNA表达变化可同时定位于单个雪旺细胞；通过mRNA和蛋白表达水平的图像分析发现，去除背根节神经元后NGF和GDNF表达在迅速下降后随即逐渐增高，但2周时仍低于共培养表达水平（P〈0．05），CNTF表达则持续下降（P〈0．01）。结论 雪旺细胞和背根节神经元共同培养后建立相互营养支持的体系，当失去神经元营养支持后，需要分泌神经生长因子和多种神经营养因子以维持自身环境的稳定；同时上述研究提供的方法能对雪旺细胞中多种神经营养因子的共表达定位进行可视化研究。     

神经干细胞移植促进脊髓损伤后脑源性神经营养因子的表达

目的：探讨神经干细胞（NSCs）移植对大鼠脊髓损伤后脑源性神经营养因子（BDNF）表达的影响及其意义。方法：NSCs提取自新生Wistar大鼠的海马区，经培养、鉴定。制作大鼠脊髓损伤（SCI）模型，于伤后第7天移植NSCs。实验分为3组：NSCs移植组（A组），DMEM填充组（B组），正常对照组（C组）。应用RT—PCR法和免疫组化法观察细胞移植后BDNF基因表达的变化。结果：RT—PCR结果分析，移植术后第1、3、5天，A组BDNF mRNA的表达量明显高于B组，差异有统计学意义（P〈0．05）。组化结果分析，移植术后第7、14、28天BDNF的表达量A组明显高于B组，差异有统计学意义（P〈0．05）。结论：NSCs在移植后可上调脑源性神经营养因子BDNF基因的表达，是其修复脊髓损伤的机制之一。     

睫状神经营养因子涂层的聚羟基乙酸聚乳酸神经导管修复犬胫神经缺损

目的 探讨应用脉冲等离子体方法涂层睫状神经营养因子（CNTF）的聚羟基乙酸聚乳酸（PGLA）神经导管修复犬胫冲经缺损的疗效。方法 18只杂交犬．每只犬左后肢制成胫神经2．5cm缺损模型，随机分别应用三种方法修复。A组：应用脉冲等离子体方法涂层CNTF的PGLA神经导管；B组：单纯PGLA神经咩管；C组：自体神经。应用苏木精-伊红和Masson染色、S-100免疫组化染色、神经电生理及神经轴突计数方法评价神经再生效果。同时动态记录犬行走步态变化，实验观察期3个月。结果 神经导管血管化良好且大部分降解吸收，再生神经均已通过所有神经导管。A组与C组的神经传导速度和神经轴突计数差异无统计学意义（P〉0．05），而A组和C组的数据均优于B组（P〈0．05）。A组和C组的犬基本恢复正常行走步态，而B组犬仍有跛行。结论 脉冲等离子体方法涂层CNTF的PGLA神经导管能有效修复犬2．5cm胫神经缺损，取得与自体神经相近的效果。     

周围神经端端或端侧吻合后降钙素基因相关肽和P物质免疫组织化学的对比研究

目的　探讨周围神经端端或端侧吻合后神经递质降钙素基因相关肽 (CGRP)和 P物质 (SP)水平的变化。　方法　选取雌性 Wistar大鼠 2 0只 ,随机分为 5组 ,每组 4只 ;实验组为 4组 ,正常对照组为 1组。实验组 :切断双侧腓总神经。将左侧腓总神经远侧断端吻合至胫神经侧方 ,右侧行腓总神经端端吻合 ,术后 1、2、4和 2 7周于各时间点分别在神经吻合口、腰段脊髓和背根神经节处取材 ,每次 4只进行 CGRP和 SP的免疫组织化学染色。对照组 :不行神经切断及吻合术 ,1周后检测同上。　结果　实验组术后 1周在腰段脊髓后角和背根神经节中 CGRP和 SP的表达均显著减少 ,术后 4、2 7周 ,CGRP和 SP在腰段脊髓后角的表达逐渐恢复至接近正常水平 ,但在背根神经节中的表达却无显著变化。端端、端侧吻合的两侧 CGRP和 SP变化的趋势一致 ,但在腰段脊髓后角的 4个时间点的样本中 ,端端吻合的表达显著高于端侧吻合。对坐骨神经进行乙酰胆碱酯酶 (Ach E)、SP、CGRP和 PGP9.5染色显示神经纤维均可通过端端与端侧神经吻合口。　结论　再生的神经纤维可以通过端侧吻合口 ,端侧吻合中感觉神经的恢复与端端吻合相似 ,但恢复的程度稍差于端端吻合。     

纳米微囊-VEGF复合体对创伤组织中VEGF、受体KDR的表达及微血管形成的影响

目的：研究纳米微囊-VEGF复合体对创伤组织中VEGF、受体KDR以及微血管表达的影响，探讨其促进损伤组织修复的作用机理。方法：将纳米微囊-VEGF复合体作用于兔耳慢性创面，利用反转录PCR方法观察各组术后1、3、7、10、14天创面组织中YEGF及KDR mRNA表达的变化；利用免疫组化染色观察创面肉芽组织中VEGF蛋白表达及微血管的变化。结果：VEGF mRNA在NW（非缺血创面组）和CW＋VEGF组（转基因组）3天后的表达水平均高于其在CW组（慢性创面组）相应时间点的表达水平，P〈0．05；KDR mRNA在NW组7～14天，CW＋VEGF组3～14天的表达水平高于CW组，P〈0．05；VEGF蛋白表达变化与mRNA表达结果基本一致；与CW组相比，7天后微血管计数在CW＋VEGF组及NW组增加显著，P〈0．05。结论：外源性VEGF基因转染导致创面中VEGF及受体KDR的表达上调，并通过增加肉芽组织中微血管的生成促进了慢性创面的愈合。     

神经纤维瘤蛋白在先天性脊柱侧凸患者成骨细胞和软骨细胞中的表达

目的：检测神经纤维瘤蛋白在先天性脊柱侧凸患者成骨细胞和软骨细胞中的表达。方法：6例先天性脊柱侧凸患者，在后路手术时取髂骨及髂骨生长板，分离、培养成骨细胞和软骨细胞，分别行碱性磷酸酶染色和甲苯胺蓝染色。逆转录-多聚酶链反应（RT—PCR）检测神经纤维瘤蛋白mRNA．间接免疫荧光和Westemblot检测神经纤维瘤蛋白在成骨细胞和软骨细胞中的表达。结果：先天性脊柱侧凸患者成骨细胞和软骨细胞中存在Ⅱ型神经纤维瘤蛋白表达，该蛋白主要分布在细胞浆，所表达蛋白为三磷酸鸟苷酶活化蛋白（GAP）活性较弱的Ⅱ型异构体。结论：先天性脊柱侧凸患者成骨细胞和软骨细胞中存在神经纤维瘤蛋白表达，但该蛋白是否通过对成骨细胞和软骨细胞的影响导致骨骼系统异常还有待于进一步研究。     

单侧隐睾睾丸组织中胰岛素样因子3基因表达

目的 研究单侧隐睾对睾丸组织胰岛素样因子3（Insl3）表达的影响。方法 通过建立隐睾大鼠动物模型获得A组单侧隐睾大鼠12只和B组双侧隐睾大鼠10只，C组空白对照正常大鼠10只。在大鼠8周龄时取睾丸组织用Tarazol法提取总RNA，Northern Blot方法检测各组睾丸组织中Insl3基因表达水平；同时提取各组睾丸组织蛋白，Westernblot检测Insl3蛋白多肽表达。结果 Insl 3 mRNA在单侧隐睾侧睾丸组织中表达量低于C组（P〈0．05），单侧隐睾大鼠对侧睾丸表达量低于C组（P〈0.05）；InSl3基因表达的蛋白多肽在隐睾侧睾丸组织中表达量较低（P〈0．05），对侧睾丸组织蛋白表达较C组低（P〈0．05），但较隐睾侧睾丸和B组明显增多（P〈0．05）。结论 单侧隐睾生精功能的损害可能是影响了隐睾侧睾丸Insl3基因表达，同时还抑制对侧睾丸的表达。     

侧侧缝合法治疗不完全性周围神经损伤的实验研究

目的： 种新的治疗不完全性周围神经损伤的方法一侧侧缝合法，并对其疗效进行初步的实验研究。方法：SD大鼠12只，双下肢随机分为实验侧和对照侧。将两侧腓总神经在相同部位以相同的力度钳夹损伤；实验侧将损伤的腓总神经远端与胫神经靠拢后，切开相邻面的束、外膜，互相侧侧缝合，对照侧不作进一步处理。3个月后，对神经的再生情况进行肌电图、组织学等检查。结果：实验侧腓总神经远端有良好的神经再生，再生的神经纤维质量与对照侧相比有显著差异（P＜0．05）。结论：不完全性周围神经损伤经侧侧缝合修复，可以获得较好的再生；侧侧缝合法是一种新的修复不完全性周围神经损伤的方法。     

脊髓腹侧压迫损伤后MP、Iloprost和Riluzole对Bcl—2表达的影响

目的：观察脊髓腹侧迫损伤后Bcl-2表达的动态变化及神经保护剂对Bcl-2表达的影响。方法：大鼠胸脊髓腹侧压迫损伤后,将受试运动分别用甲基强的松龙（MP）、Iloprost,Riluzole及以上三种药物合用联合治疗。应用免疫组织化学及原位杂交方法检测治疗组及损伤组中Bcl-2的表达。结果：脊髓伤后1h Bcl-2蛋白增加,伤后3h达高峰,伤后2周表达接近于正常水平。Bcl-2 mRNA的表达也有相似的变化过程。。MP、Iloprost和Riluzole能增加Bcl-2蛋白表达,但对Bcl-2 mRNA的表达增加不明显。结论：作用方式不同的几种神经保护剂均能增加脊髓损伤组织中的Bcl-2蛋白的表达。     

兔坐骨神经损伤修复后骶交感神经节内c-ret mRNA表达的变化

目的 探讨兔坐骨神经损伤修复后骶交感神经节内c-ret mRNA表达的变化。方法 用组织原位杂交方法观察兔坐骨神经损伤修复1，3，7，14，30，60d后c-ret mRNA在骶交感神经节内的表达。结果 兔坐骨神经损伤修复3d后，在损伤侧骶交感神经节内c-ret mRNA表达明显上调，在损伤后7d达高峰，损伤后14d与30d仍维持较高水平，损伤后60d基本恢复到正常水平。结论 C—ret mRNA在兔坐骨神经损伤修复后骶交感神经节内表达的变化，提示在坐骨神经损伤修复后给予外源性胶质源性神经营养因子(GDNF)可能有利于交感神经纤维的再生。     

An updated review on the principles of intraoperative neurophysiological monitoring and the anaesthetic considerations

It is known that any surgery to the nervous system poses risks to neural structures and their surrounding structures. These mechanisms of injury are the result of mechanical manipulations, haemodynamic alterations, chemical or thermal injuries. Intraoperative neurophysiological monitoring (IONM), using various modalities, is employed to facilitate the assessment of the functional integrity of neural structures, and it is used to provide a real-time alerting system when changes caused by surgically induced insults are detected. The primary goal of IONM is reducing the risk of postoperative neurological deficits during these surgical procedures. It is used to provide information that allows the surgeon to correct any surgical interventions that may have compromised these systems and this also in turn provides guidance on what neurological deficits to anticipate postoperatively. Apart from being utilized as an alerting system to avoid catastrophic outcomes, IONM also assists as a guidance system using stimulation techniques to map out eloquent areas within the cortex, allowing identification of specific neuronal structures, particularly when landmarks cannot be easily recognized due to infiltration by pathological lesions.In this article, we focus on updating our previous paper published in 2019 and again, to provide attention to the various neurophysiological modalities that are employed in IONM. We will look at the basic underlying physiological principles and their individual indications for use clinically. We will explain the information that each modality provides. Importantly, and the primary reason for this article, we look at the various anaesthetic agents, their effects on each neurophysiological modality and other anaesthetic considerations such as haemodynamic and temperature effects. We will also recommend the use of an alert checklist for the multidisciplinary team should an intraoperative alert be issued during surgical procedures.     

中医骨伤科学的现代化是可持续发展的必由之路

21世纪，现代科学和现代医学高速发展，中医面临着机遇与挑战。随着人类对健康需求的不断增强和深化，中医已成为世界范围内医学发展关注的热点。而在国内，随着经济体制和社会保障改革的深化，中医产业化也被纳入到国家经济发展规划当中，可以说，新世纪中医飞速发展的大气候已经形成。当前乃至今后，中医的发展必将是国际化的。中医骨伤科学是中医学中一个颇具特色和优势的分支，其发展也必将是国际化的。     

The role of the transient receptor potential (TRP) superfamily of cation-selective channels in the management of the overactive bladder

? The pathophysiology of lower urinary tract symptoms (LUTS), detrusor overactivity (DO), and the overactive bladder (OAB) syndrome is multifactorial and remains poorly understood. ? The transient receptor potential (TRP) channel superfamily has been shown to be involved in nociception and mechanosensory transduction in various organ systems, and studies of the LUT have indicated that several TRP channels, including TRPV1, TRPV2, TRPV4, TRPM8, and TRPA1, are expressed in the bladder, and may act as sensors of stretch and/or chemical irritation. ? However, the roles of these individual channels for normal LUT function and in LUTS/DO/OAB, have not been established. ? TRPV1 is the channel best investigated. It is widely distributed in LUT structures, but despite extensive information on morphology and function in animal models, the role of this channel in normal human bladder function is still controversial. Conversely, its role in the pathophysiology and treatment of particularly neurogenic DO is well established. ? TRPV1 is co-expressed with TRPA1, and TRPA1 is known to be present on capsaicin-sensitive primary sensory neurones. Activation of this channel can induce DO in animal models. ? TRPV4 is a Ca(2+)-permeable stretch-activated cation channel, involved in stretch-induced ATP release, and TRPV4-deficient mice exhibit abnormal frequencies of voiding and non-voiding contractions in cystometric experiments. ? TRPM8 is a cool receptor expressed in the urothelium and suburothelial sensory fibres. It has been implicated in the bladder-cooling reflex and in idiopathic DO. ? The occurrence of other members of the TRP superfamily in the LUT has been reported, but information on their effects on LUT functions is scarce. There seem to be several links between activation of different members of the TRP superfamily and LUTS/DO/OAB, and further exploration of the involvement of these channels in LUT function, normally and in dysfunction, may be rewarding.     

电视胸腔镜手术在肺硬化性血管瘤诊断和治疗中的应用

目的探讨电视胸腔镜手术在以孤立性肺结节（solitary pulmonary nodule,SPN）为表现的肺硬化性血管瘤（pulmonary sclerosing hemangioma,PSH）诊断和治疗中的价值。方法回顾性分析1999年2月-2008年10月,11例术前经胸片、胸部CT发现的周围型SPN（咳嗽2例,咳血1例,胸痛1例,无症状7例）,术后病理确诊为肺硬化性血管瘤的临床资料。全胸腔镜或胸腔镜辅助小切口肺楔形切除7例,肺段切除3例,肿瘤切除术1例。结果 11例均成功完成电视胸腔镜手术（video-assisted thoracoscopic surgery,VATS）。术中快速冰冻病理示PSH7例,2例肺良性病变,2例恶性肿瘤待排除;术后病理确诊为PSH。平均手术时间57 min（45-70 min）,平均出血量85 ml（50-120 ml）;术后无并发症和手术死亡。11例术后随访22-63个月,平均31个月,无复发。结论 PSH术前诊断困难,电视胸腔镜不仅是PSH的诊断手段,也是目前最有效的治疗方法。     

The role of surgical transcatheter valve implantation for the treatment of severe mitral annular calcification

With a rise in the aging population, mitral annular calcification is increasingly encountered with an incidence of 10% in over 70 years old. This with increasing patient comorbidities presents a technical challenge due to the risk of atrioventricular disruption which is associated with high operative mortality of up to 75%. We describe two cases of severe mitral disease with marked annular calcification successfully treated with a balloon expandable transcatheter valve which was deployed on cardiopulmonary bypass via a trans‐atrial approach.     

Role of CT and MRI in the design and development of orthopaedic model using additive manufacturing

Objective

To study the role of Computed tomography (CT) and Magnetic resonance imaging (MRI) for design and development of orthopaedic model using additive manufacturing (AM) technologies.

Evaluating the safety of the Harmonic Scalpel around the recurrent laryngeal nerve

Background: The Harmonic Scalpel (HS) is widely used in thyroidectomy. Determining the safety margin of using the HS near the recurrent laryngeal nerve (RLN) is helpful in preventing the injury to this nerve during thyroidectomy. Methods: The parameters of evoked electromyography (EEMG) of vocal muscles before and after using the HS at a power level of 3 were recorded in a rabbit model. Masson staining was used to determine lateral injury caused by incisions using the HS. Results: After the activated tip of the HS made contact with the RLN for ≥1 s or was placed 1 mm from the nerve for 3 or 5 s or 2 mm from the nerve for 5 s, significant changes were observed in the minimal stimulus current intensity threshold, the optimal stimulus current intensity threshold, the onset latency and the wave amplitude of EEMG. After the activated HS tip touched the RLN or was placed 1 mm to the nerve for ≥1 s or 2 mm to the nerve for 5 s, significant changes were found in peak latency. The thickness of injured lateral tissue was <1, 1–2 and >2 mm when using HS for 1, 3 and 5 s, respectively. Conclusion: When used near the RLN at a power level of 3, the activated HS tip should be ≥2 mm from the nerve and the duration of incision should be ≤3 s.     

神经节苷脂有髓损伤的保护作用观察

目的：观察神经节苷脂对损伤脊髓神经组织的保护作用,并初步探讨其作用机制。方法：重度脊髓损伤大鼠随机分为对照组及神经节苷脂治疗组（n=15)。在蛛网膜下腔注射生理盐水20μl或神经节苷脂30μg。观察体重变化局部血流量、凋亡细胞原位标记、神经功能评价和定量组织学分析。结果：局部血流量2组无明显差异,治疗组存活8率、体重、神经功能评分和残余神经组织面积明显高于对照组,而TUNEL阳性细胞数量明显低于对照组。结论：神经节苷脂对损伤脊髓组织有明显保护作用,可能是通过对神经细胞凋亡的阻断来发挥作用的。