带蒂骨骼肌桥接近端周围神经防治残端痛性神经瘤的实验与临床应用研究

目的：探讨神经瘤的形成机制和致痛机制及周围神经与带蒂骨骼肌桥接吻合后神经的生长情况。方法：Wistar大白鼠40只随机分为实验组与对照组，两组均将左侧坐骨神经在适当水平切断，远端神经切除，实验组近端神经分为2束，用带蒂骨骼肌桥接2个神经断端，对照组近端神经置于原位。饲养16周进行组织学检测。临床应用此方法治疗残端痛性神经瘤24例。结果：实验组再生的神经纤维顺利通过吻合口长入骨骼肌桥内，并在肌桥的肌束内生长，分布于肌纤维之间，未见有神经瘤形成。对照组均有典型的神经瘤形成。临床24例病人总计36个神经瘤，随访6～20个月按中日联谊医院评定标准优良率为92％。结论：带蒂骨骼肌桥接近端周围神经是防治残端痛性神经瘤的一种简单、实用、有效的方法。     

同侧颈7神经根移位治疗臂丛上干根性撕脱伤患肢功能变化的实验研究

目的研究臂丛上干根性撕脱伤再切取同侧颈，后不造成肢体功能进一步损害的代偿机制。方法60只SD大鼠随机等分为二组，上干根性撕脱＋同侧颈7神经根切断为实验组，单纯上干根性撕脱为对照组。术后3、6和12周每组取10只大鼠对颈7神经根代表肌（背阔肌、肱三头肌、桡侧腕短伸肌和指总伸肌）作肌肉组织学、神经电生理与运动终板组织形态学的检测。结果术后3周实验组与对照组相比，四块颈7代表肌的湿重显著减轻且肌细胞截面积缩小，复合肌肉动作电位（CMAP）的潜伏期显著延长并波幅明显下降，运动终板突触后膜面积明显减小，上述指标的二组差异均有统计学意义（P〈0．05或P〈0．01）。荧光显微镜观察显示实验组颈7代表肌的运动终板分布稀疏，电镜下可见形态不成熟的再生运动终板。伤后6周，实验组肱三头肌CMAP潜伏期及波幅的恢复率以及桡侧腕短伸肌与指总伸肌肌细胞截面积、CMAP潜伏期及波幅和突触后膜面积的恢复率与对照组相比，差异无统计学意义（P均〉0．05）。实验组颈7代表肌的运动终板数量已较3周时明显增多，超微形态亦趋于成熟。到12周，实验组各项检测指标的恢复率均已接近对照组（P均〉0．05），其颈7代表肌的运动终板分布密集且数量较多，超微结构接近成熟，形态与对照组基本一致。结论臂丛上干根性撕脱伤切取同侧颈7神经根后，下干神经纤维能通过末梢运动终板的再生对中干代表肌进行代偿性支配，故不会导致肢体功能的进一步损害。     

中药治疗对大鼠坐骨神经损伤后神经生长因子蛋白表达的影响

目的：研究党参、丹参、黄芪、生地等单味中药及复方神肌再生冲剂对大鼠坐骨神经损伤后神经生长因子（nerve growth factor,NGF)蛋白表达的影响。方法：雄性SD大鼠108只，按手术先后随机分成党参、丹参、黄芪、生地、复方神肌再生冲剂和对照组6组，每组18只。按取材时间分为术后2、4、8、周3个时间组（每组6只）。显露大鼠右侧坐骨神经，于坐骨结节远端0.8cm处造成坐骨神经挤压伤。根据分坐骨神经干，应用免疫组织化学和图像分析的方法研究神经组织中NGF的表达并进行定量分析。结果：术后4、8周组，丹参和复方神肌再生冲剂组，坐骨神经组织中的NGF蛋白高表达。结论：大鼠坐骨神经损伤后用中药丹参和复方神肌再生冲剂治疗，可促进坐骨神经组织NGF蛋白表达。     

FK 506加速周围神经损伤修复后的功能恢复

目的：探讨FK506对大鼠坐骨神经横断伤修复后肢体功能恢复的影响。方法：45只SD大鼠高位切断坐骨神经后原位缝合，术后实验组（25只）用FK506灌胃，对照组（20只）不给药，于术后第1、2、3、5个月检测术肢比目鱼肌肌湿重恢复率，小腿三头肌肌力恢复率，坐骨神经功能指数及皮层体感诱发电位（SEEP）的潜伏期，结果：实验组肢体功能恢复时间较对照组提前大约2个月。结论：FK506可加速周围神经横断伤修复后肢体功能的恢复。     

FK506加速周围神经损伤修复后肢体功能恢复的实验研究

目的：探讨FK506对大鼠坐骨神经横断伤修复后肢体功能恢复的影响。方法：SD大鼠高位切断坐骨神经后原位缝合，术后实验组FK506灌胃,对照组不给药。分别于第1、2、3、5月检测术肢比目鱼肌重量恢复率、小腿三头肌肌力恢复率、坐骨功能指数、皮层体感诱发电位潜伏期。结果：实验组肢体功能恢复时间较对照组提前大约两个月。结论：FK506可加速周围神经横断伤后肢体功能恢复。     

臂丛损伤后神经移位寄养法预防失神经支配肌萎缩的实验研究

目的研究臂丛损伤后神经移位端侧缝合寄养法是否有预防失神经支配骨骼肌肌萎缩的作用。方法64只SD大鼠随机分成两组。对照组：切断肌皮神经，造成肱二头肌失神经支配。实验组：切断肌皮神经后，胸内侧神经分支移位与肌皮神经远端作端侧缝合寄养失神经支配的肱二头肌。术后2、4、6、8周观察大鼠的行为变化与肱二头肌的萎缩程度，检测肱二头肌肌肉纤颤电位或再生电位、肱二头肌肌肉湿重、肌纤维截面积和Na-K-ATP酶活性。以左侧为实验侧，右侧为自身对照侧，将左侧测量值除以右侧测量值，求各观察值恢复率，比较组间各观察值的恢复率。结果术后对照组随着失神经时间延长，肌肉萎缩程度逐渐加重，屈肘功能不能恢复，纤颤电位波幅逐渐下降，肌肉湿重、肌纤维截面积和酶的活性均逐渐下降；而实验组随着神经寄养时间的延长，肌肉萎缩程度逐渐减轻，屈肘功能逐渐恢复，出现再生电位，肌肉湿重、肌纤维截面积逐渐增加，酶活性逐渐升高，虽不及正常组，但明显不同于肌肉萎缩严重的失神经组。结论神经移位端侧缝合寄养法可以有效地预防失神经支配骨骼肌肌萎缩。     

神经干细胞移植再支配失神经骨骼肌的实验研究

目的 探讨采用神经干细胞移植的方法使失神经骨骼肌重获神经再支配的可行性。方法 将108只SD大鼠按注射药物的不同随机分为3组，每组36只大鼠。切断右侧胫神经建立腓肠肌失神经实验模型。实验组：将神经干细胞悬液注射到切断胫神经的远端。对照组：注射等量的细胞培养液。损伤组：注射等量的生理盐水。术后8、10、12周采用免疫组织化学、HRP逆行示踪技术和神经形态学的方法，检测失神经骨骼肌是否重新获得移植神经干细胞的再支配。结果 术后8周局部即可检测到分化的神经元和神经胶质细胞，其再生的轴突与靶肌肉已经建立神经突触连接。术后10、12周时，再生的神经纤维进一步增多。结论 周围神经断伤后局部用神经干细胞移植能够使远端失神经骨骼肌重新获得神经再支配。     

靶肌肉注射甲状腺素促进周围神经再生的实验研究

目的研究在靶肌肉注射甲状腺素(T3)对周围神经再生的作用。方法将60只大鼠随机分为实验组和对照组,每组30只。硅胶管桥接大鼠坐骨神经,实验组于术后每周2次将甲状腺素注射于伤侧胫前肌和腓肠肌上,术后1个月和3个月,应用运动神经传导速度(MNCV)、光镜、透射电镜等方法分别从功能和形态方面测定各项指标;对照组注射等量的生理盐水。结果实验组再生神经在功能和形态方面均优于对照组。结论靶肌肉注射甲状腺素能有效促进周围神经再生。     

缺血预处理改善骨骼肌功能的研究

目的：探讨缺血预处理法改善缺血骨骼肌功能的临床价值。方法：用ＳＤ大鼠１２只，以右后肢为动物实验模型。分为缺血组（对照组，鼠６只），即缺血４小时后再灌注１小时的方法；缺血预处理组（实验组，鼠６只），缺血过程同对照组，但在缺血前预先经过２次缺血５分钟、再灌注１０分钟的处理。实验时，分别于缺血前、缺血１、４小时及再灌注１小时时，测定两组实验侧肢体腓肠肌最大肌张力的变化。实验结束后分别测量血ＭＤＡ、ＣＰＫ及大鼠右后肢９９ｍＴｃ亚甲基二磷酸计数。结果：实验组最大肌张力的变化（缺血４小时、再灌注１小时时）较对照组有明显改善；血ＭＤＡ、ＣＰＫ及肌肉９９ｍＴｃ亚甲基二磷酸较对照组显著降低。结论：缺血预处理不仅能改善骨骼肌的缺血耐受性，而且能有效地改善骨骼肌的功能     

化学萃取自体骨骼肌桥修复神经缺损初步研究

目的 观察化学萃取自体骨骼肌桥修复大鼠坐骨神经缺损的可行性。方法 用化学萃取方法制备自体肌骼肌的无细胞基底膜管，修复大鼠坐骨神经１５ｍｍ神经缺损，与冻融肌作骼肌桥作对照，用免疫组织化学方法及碱性磷酸酶组织化学方法观察神经及血管再生的情况。结果 化学萃取自体骨骼肌桥在修复神经缺损伤后，其物理性能，移植物血管化过程和神经再生速度与质量均优于冻融肌桥。     

中药对小鼠骨折后血管重建与骨折愈合方式关系的影响

目的 探讨中药对小鼠骨折后血管重建与骨折愈合方式关系的影响。方法 昆明种小鼠50只，制作闭合性胫腓骨骨折模型，随机分为两组，实验组口服中药，对照组口服生理盐水。分别于术后第3、7、15、21、28天处死动物。腹主动脉墨汁灌注，制成石蜡切片。HE染色，光镜观察。结果 实验组骨折部位血管的增生、扩张程度高于对照组。软骨内成骨占整个骨痂的比例少于对照组。实验组较对照组约提前1周愈合。结论 中药通过促进骨折部位血管增生、扩张，加速软骨骨痂改建，减少软骨内成骨，增加膜内成骨的比例，促进骨折的愈合。     

外敷中药对骨折愈合微血管重建的影响

目的 探讨中药对骨折愈合微血管重建的影响。方法 家兔１５只,双侧胫骨制作骨折模型,左侧外敷中药,右侧未用中药为对照组。术后１０,２０,３０天分别处死动物,将中国墨汁注射入腹主动脉,采用组织切片法,观察微血管变化,以评价中药对骨折愈合的影响。     

冬虫夏草对缺血再灌注肢体能量代谢和线粒体酶活性的影响

目的 观察冬虫草醇提取液对肢体缺血再灌注损伤的影响。方法 制作兔肢体缺血再灌注损伤动物模型，实验分对照组、再灌注组和治疗组。取骨骼肌测定三磷酸腺苷、二磷酸腺苷、一磷酸腺苷、磷酸肌酸含量和线粒体ATP酶活性。结果 再灌注组与对照组比较，骨骼肌能量代谢障碍及其线粒体ATP酶活性降低。使用冬虫夏草后，骨骼肌各项测定指标较再灌注组相比明显改善。结论 冬虫夏草对缺血再灌注损伤骨骼肌有保护作用。     

恒磁场对SD大鼠深创面愈合的影响

目的：探讨恒磁场对SD大鼠深创面愈合的影响,恒磁场对创面愈合过程中VEGF表达的影响,以及不同强度恒磁场治疗SD大鼠深创面的区别。方法：将48只清洁级SD大鼠随机分成0．16T、0．32T磁疗组和对照组。分别在术后第3、6、9、12天每组处死4只大鼠,测定创面愈合指数,免疫组化检测肉芽组织VEGF表达。比较各组的创面愈合指数及VEGF表达情况。结果：④愈合指数。0．16T磁疗组在第6、9天时的愈合指数高于对照组,差异有统计学意义;0．32T磁疗组在第3、6、9、12天时的愈合指数高于对照组,差异有统计学意义。0.32T磁疗组与0．16T磁疗组相比差异无统计学意义。②创面修复不同时期VEGF表达观察结果。在术后第3、6天磁疗组VEGF表达强于对照组,差异有统计学意义;第9、12天磁疗组与对照组相比差异无统计学意义（P〉0．05）。0．32T磁疗组与0．16T磁疗组相比差异无统计学意义。两个磁疗组在第6天时VEGF阳性率达到整个愈合过程中的峰值,而对照组在第9天时才达到高峰,且VEGF表达强度低于两个磁疗组。结论：0．16T及0．32T恒磁片均能促进SD大鼠深创面的愈合,恒磁场促进深创面愈合的机制可能和在创面愈合早、中期增强VEGF的表达有关。     

腰椎间盘突出症患者髓核组织中受体相互作用的丝氨酸苏氨酸激酶1表达和临床意义

目的:探讨腰椎间盘突出症(lumbar disc herniation,LDH)患者髓核组织中受体相互作用的丝氨酸苏氨酸激酶1(receptor-interacting protein serine-threonine kinases 1,RIPK1)表达和临床意义。方法:选取2016年1月至2018年1月治疗40例LDH患者的髓核组织标本为病例组,另选取同期行手术治疗30例腰椎骨折患者的髓核组织为对照组,分别采用聚合酶链反应(polymerase chain reaction,PCR),Western blot法检测两组患者髓核组织中RIPK1 mRNA和蛋白的表达,采用免疫组织化学染色法检测两组患者髓核组织中RIPK1蛋白表达,采用ELISA法检测两组患者髓核组织中RIPK1和TNF-α浓度,采用单因素方差分析髓核组织中RIPK1和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)浓度与LDH患者Pearce分级间的关系,采用Pearson分析RIPK1与TNF-α浓度间的相关性。结果:RIPK1在对照组髓核组织中呈弱阳性表达,RIPK1蛋白在病例组中呈阳性或强阳性表达。病例组髓核组织RIPK1 mRNA表达量高于对照组(P<0.05)。病例组髓核组织中RIPK1蛋白表达量高于对照组(P<0.05)。病例组髓核组织RIPK1浓度高于对照组(P=0.012)。病例组髓核组织TNF-α浓度显著高于对照组(P=0.009)。不同Pearce分级的LDH患者髓核组织中RIPK1和TNF-α浓度存在显著性差异(P>0.05),髓核组织中RIPK1和TNF-α浓度随着Pearce分级增加而显著升高。Pearson相关分析示LDH患者髓核组织中RIPK1和TNF-α浓度间存在显著正相关(r=0.781,P<0.001)。结论:LDH患者椎间盘组织中RIPK1 mRNA和蛋白表达水平高于正常的椎间盘组织,随着Pearce分级增高而升高,其可能是参与LDH炎性病变的重要因子。     

Effects of CO2 insufflation and laparotomy on wound healing in mice

BACKGROUND: The aim of the present study was to evaluate the effects of laparotomy and CO2 insufflation on wound healing in a murine incisional wound healing model. METHODS: Seventy-two male Swiss Albino mice were randomly allocated into three groups of control, laparotomy and CO2 insufflation. A transverse skin incision of 15 mm was made in the dorsum of each mouse, and four interrupted mattress sutures with 4.0 polypropylene thread were laid for wound closure. A median laparotomy was performed in the laparotomy group. CO2 insufflation was performed with an intra-abdominal pressure of 9 mmHg. The retained gas was evacuated from the abdominal cavity at the end of a 60-min period. Mice were killed on the 3rd, 7th and 15th postoperative days. The wound tensile strength and 5-hydroxyproline concentration in the wound tissue were measured. RESULTS: Tensile strength of the incised skin increased as the post-incision period progressed. There was no significant difference between the tensile strengths of the incised skin of control, laparotomy and CO2 insufflation groups throughout the observation period. The skin 5-hydroxyproline concentrations of all groups were not significantly different at the 3rd postoperative day. But laparotomy and CO2 insufflation groups had lower 5-hydroxyproline concentrations at the 7th and 15th postoperative days, when compared to controls (P < 0.02 for 7th and 15th days). CONCLUSION: CO2 insufflation and laparotomy reduce the 5-hydroxyproline concentration of the wound, suggesting a diminished wound healing capacity.     

Book reviews

Background and purpose Animal models of skeletal muscle injury should be thoroughly described and should mimic the clinical situation. We established a model of a critical size crush injury of the soleus muscle in rats. The aim was to describe the time course of skeletal muscle regeneration using mechanical, histological, and magnetic resonance (MR) tomographic methods.

Methods Left soleus muscles of 36 Sprague-Dawley rats were crushed in situ in a standardized manner. We scanned the lower legs of 6 animals by 7-tesla MR one week, 4 weeks, and 8 weeks after trauma. Regeneration was evaluated at these times by in vivo measurement of muscle contraction forces after fast-twitch and tetanic stimulation (groups 1W, 4W, 8W; 6 per group). Histological and immunohistological analysis was performed and the amount of fibrosis within the injured muscles was determined histomorphologically.

Results MR signals of the traumatized soleus muscles showed a clear time course concerning microstructure and T1 and T2 signal intensity. Newly developed neural endplates and myotendinous junctions could be seen in the injured zones of the soleus. Tetanic force increased continuously, starting at 23% (SD 4) of the control side (p < 0.001) 1 week after trauma and recovering to 55% (SD 23) after 8 weeks. Fibrotic tissue occupied 40% (SD 4) of the traumatized muscles after the first week, decreased to approximately 25% after 4 weeks, and remained at this value until 8 weeks.

Interpretation At both the functional level and the morphological level, skeletal muscle regeneration follows a distinct time course. Our trauma model allows investigation of muscle regeneration after a standardized injury to muscle fibers.     

封闭负压引流早期处理软组织爆炸伤创面的实验研究

目的：观察封闭负压引流（vacuum assisted closure,VAC）早期处理对软组织爆炸伤创面的影响,为战伤早期处理提供新的思路。方法：选取健康3月龄长白猪8头,体重（50±5）kg,雌雄不拘,用同一型号的电雷管（型号：660929F48840-55,含DDNP0.3g,RDX0.7g）紧贴双侧臀部皮肤,造成16个损伤程度相当的爆炸创面,左右创面配对分成实验组和对照组。在室外暴露3h后进行创面的彻底清创,实验组用封闭负压引流处理,负压值维持在（-50±5）Kpa;对照组用无菌干纱布覆盖。分别于治疗前后进行大体观察、细菌学计数和革兰氏阳性（G＋）细菌比例分析,以及采集病理学标本进行HE染色和Masson染色。结果：治疗3d后实验组创面细菌数为[（7.82±0.55）×104]CFU/g,对照组为[（1.07±0.14）×106]CFU/g,两组间有统计学差异（P〈0.01）;实验组G＋细菌比例显著提高,对照组则没有变化;实验组创面清洁,较对照组有更丰富的新生肉芽组织、血管和胶原,坏死组织明显减少。结论：VAC能抑制创面细菌增殖,提高G＋细菌的比例,促进创面愈合,可用于软组织爆炸创面的早期治疗。     

The effect of mesenchymal stem cell lyophilizate on the recovery of the zone of stasis following thermal burns

Introduction and aimMesenchymal Stem Cells (MSCs) are known to contribute to wound healing by increasing tissue regeneration. This study examined the effect of MSC-Lyophilizate (MSC-L) on the recovery of the zone of stasis in thermal burns.MethodsA comb was used to induce second-degree thermal burns (1 × 2 cm) on the dorsum of the rats. Within 30 min after the burn, MSC-L derived from the umbilical cord was administered to the experimental group and 1.5 ml of 0.9% isotonic sodium chloride to the sham group. The control group did not receive any intervention. Tissue samples were collected on postoperative day 7. Histopathological assessments were made using a microscope with digital camera attachment. SPSS for IBM 25 was used for data analysis.ResultsEpithelial loss and subepidermal bullae were observed in the control and sham groups on day 7. In the experimental group, the MSC-L administration was found to increase epithelial tissue formation and neovascularization in the dermis. We found no significant pathological findings in the epidermis and dermis in the experimental group.ConclusionAdministration of umbilical cord-derived MSC-L is of potential importance in wound healing. In our study, we observed that MSC-L that contained 1.5 million cells contributed significantly to the recovery of the stasis zone of burn.     

Neuronal regeneration in denervated muscle following sensory and muscular neurotization

Background and purpose?Neurotization of denervated muscles has been shown to improve muscle bulk, but the neuronal regeneration response has not been compared previously in different surgical techniques of neurotization. Thus, using a rat model of experimental skeletal muscle denervation, we studied neuronal regeneration following sensory neurotization by two methods: sensory nerve to motor branch of muscle and direct sensory nerve implantation to muscle.

Material and methods?The lateral head of the gas-trocnemius muscle was denervated in 36 rats, of which the first 12 served as denervated controls. In the second group of 12, the sural nerve was anastomozed to the motor branch of the gastrocnemius muscle (sensory-to-motor nerve neurotization) and in the remaining 12 rats the sural nerve was split into 4 fascicles and embedded into 4 quadrants of the muscle (direct sensory nerve-to-muscle neurotization). Immunohistochemistry was used to examine nerve fibers in muscle containing the sensory neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP), and general neuronal marker protein gene product 9.5 (PGP 9.5).

Results?Semiquantitative analysis showed that, compared to the control side, the number of nerve fibers on the experimental side was highest (p < 0.01) for group III (direct sensory nerve-to-muscle neurotization) for all 3 markers. The difference was 71%, 298%, and 254% for PGP 9.5, CGRP, and SP, respectively.

Interpretation?This method may be a good option for inducing neuronal regeneration in denervated muscles, and has therapeutic implications for prevention of atrophy of denervated muscles and as an adjunct for reconstruction of soft tissue defects.