膀胱二聚体Cajal间质细胞的超微结构及其对ATP的电反应

目的:为证明膀胱二聚体Cajal间质细胞(interstitial cells of Cajal,ICC)发挥起搏作用提供超微结构和功能学证据。方法:电子显微镜下鉴定组织和培养的豚鼠膀胱二聚体ICC并观察ICC之间的超微联系;体外分离、培养豚鼠膀胱ICC,运用细胞膜片钳技术观察ATP(100μmol/L)刺激下二聚体ICC和单体ICC的兴奋性差别。结果:二聚体ICC中两个ICC细胞胞体紧密相连,细胞之间的间隙小于20 nm,存在大量缝隙连接,局部细胞膜融合,形成联胞体。在ATP(100μmol/L)的刺激下体外培养的二聚体ICC的内向电流幅度为286.9±26.4 pA,显著大于单体ICC的内向电流幅度163.8±18.6 pA(P<0.01)。结论:二聚体ICC具有形成高兴奋起搏电流的超微结构基础,高兴奋性的二聚体ICC可能才是逼尿肌的起搏细胞。     

豚鼠输尿管Cajal间质细胞毒蕈碱受体的表达及功能研究

目的研究毒蕈碱受体在豚鼠输尿管Cajal间质细胞（ICC）上的表达,初步探讨ICC在胆碱能神经递质传递中的作用。方法将20只豚鼠随机分成对照组和实验组,进行输尿管肌条实验,观察在卡巴胆碱作用下输尿管肌条的收缩幅度和频率的变化;10只豚鼠,将输尿管进行固定、撕片和荧光染色,检测豚鼠输尿管毒蕈碱受体亚型（M1-M5）在ICC上的表达情况。结果卡巴胆碱作用后,实验组肌条的收缩幅度为0 g,频率为0次/分,对照组的幅度为（0.106±0.021）g,频率为（4.900±0.875）次/分,两组差异显著（P〈0.01）。免疫荧光双标显示,输尿管ICC仅表达M2、M3受体亚型。结论胆碱能神经可能通过ICC来调控输尿管平滑肌。     

糖尿病豚鼠膀胱Cajal样间质细胞形态学变化及意义

目的 观察糖尿病豚鼠膀胱Cajal样间质细胞形态学变化,探讨糖尿病膀胱功能改变原因.方法 用链脲佐菌素单次注射建立豚鼠糖尿病模型,建模成功4周后,行尿动力学检查,采用组织冷冻切片、酪氨酸蛋白激酶受体单克隆抗体间接免疫荧光检测及透射电镜技术观察膀胱中Cajal样间质细胞变化.结果 与对照组相比,糖尿病豚鼠膀胱最大逼尿肌压显著降低,最大膀胱容量及膀胱顺应性显著明显增高.免疫荧光检测显示逼尿肌中KIT阳性细胞与逼尿肌组织面积百分比均明显小于对照组.电镜下可见Cajal样间质细胞内细胞器变性、减少,胞质广泛溶解,核周间隙增宽,与其他细胞间的缝隙连接显著减少、结构破坏、连接松散.结论 糖尿病豚鼠膀胱Cajal样间质细胞的数量减少和结构破坏可能是造成膀胱功能障碍的重要原因之一.     

c-kit阳性细胞和胎儿食管平滑肌发育的关系

目的:探讨c-kit阳性细胞和α-平滑肌肌动蛋白阳性平滑肌细胞(α-SMA)在人胎儿食管中分布关系.方法:采用免疫荧光组织化学双重标记法对8例胎儿食管中c-kit阳性细胞和α-SMA阳性平滑肌细胞的分布进行观察.结果:c-kit阳性细胞在食管上段内肌层内侧有少量分布,肌间层和黏膜下层也可见零星分布,在食管中段外肌层开始有少量分布,内肌层数量中等,至食管下段内外肌层均有大量分布.α-SMA阳性平滑肌细胞在食管肌层由上至下逐渐增加,并由内肌层扩展到外肌层.在食管上端肌间层和黏膜下层可见少量c-kit和α-SMA共同表达.结论:c-kit阳性细胞可能是Cajal间质细胞,在胎儿食管的分布与平滑肌密切相关.食管内平滑肌细胞和Cajal间质细胞可能起源于同一种前体细胞.     

改进型“瑞士卷”法在小鼠肠道组织切片中的应用

目的为提高小鼠肠管组织石蜡切片的整体质量, 探讨一种改进型"瑞士卷"法在肠道组织病理切片中的应用。方法分别采用常规"瑞士卷"制作方法和改进型"瑞士卷"法脱水包埋小鼠回肠、结直肠组织样本, 制成组织切片。HE染色用于评估组织切片整体形态质量、组织器械性损伤及黏膜下层与肌层分离的情况;免疫组织化学染色LYVE-1标记淋巴管、染色CD31标记血管, 分析黏膜下层结构完整性;采用免疫荧光染色对小鼠回肠潘氏细胞(溶菌酶阳性)及结直肠杯状细胞(MUC2阳性)予以标记。结果 HE染色显示改进型"瑞士卷"组小鼠肠管组织切片整体形态及外观更加规整, 组织未发生器械性损伤, 且极少见黏膜下层与肌层分离的现象;免疫组织化学结果显示, 改进型"瑞士卷"组小鼠肠管组织切片黏膜下层多见完整的淋巴管和血管结构, 计数其黏膜下层完整淋巴管和血管的数量分别为(701.0±46.7)条和(881.6±58.7)条, 显著高于常规方法组(149.5±23.6)条和(122.9±20.2)条(P<0.05);免疫荧光发现, 改进型"瑞士卷"组小鼠回肠潘氏细胞及结直肠杯状细胞定位准确, 极少见细胞结构缺失的情况。结论改进...     

豚鼠前列腺Cajal间质细胞的超微结构特点及功能

目的:观察豚鼠前列腺Cajal间质细胞(interstitial cells of Cajal,ICCs)的超微结构特点以及ICCs和前列腺神经及平滑肌细胞之间的超微结构联系。方法:制作豚鼠前列腺组织切片并进行免疫荧光染色,使用酪氨酸激酶受体(c-Kit)抗体标记ICCs。制作豚鼠前列腺超薄组织切片,在透射电镜下寻找ICCs并观察其超微结构特点以及与周围神经、平滑肌细胞之间的超微结构联系。结果:免疫荧光标记发现豚鼠前列腺间质内有较多的c-Kit阳性ICCs。电镜下发现豚鼠前列腺组织内存在和c-Kit阳性ICCs形态、分布一致,符合典型胃肠道ICCs超微结构特点的间质细胞,这些前列腺ICCs分布于平滑肌细胞之间,与相邻的神经末梢形成典型的突触样连接,和周围的平滑肌细胞之间紧密相连。结论:豚鼠前列腺ICCs具有介导前列腺神经信号,调控平滑肌活动的超微形态学基础。     

长爪沙鼠胃肠道Cajal间质细胞的形态学

目的 探讨长爪沙鼠胃肠道Cajal间质细胞（ICCs）的形态和分布规律。 方法 采用10只成年长爪沙鼠,体重50~70g,取胃、小肠、结肠制作冷冻切片,结合全层铺片的c-Kit免疫荧光染色。结果 ICCs呈网络状分布于整个胃肠道,不同部位ICCs的分布及形态有所不同。在胃底部,仅见肌内ICCs(ICC-IM）,而在胃体和胃窦部除ICC-IM外,可见肌间ICCs(ICC-MY)分布在肌间神经丛周围;其细胞密度胃底ICC-IM最多,由胃底至胃窦逐渐减少,而ICC-MY由胃体至胃窦逐渐增多。在小肠可见ICC-IM, ICC-MY和深肌层ICCs(ICC-DMP)3个亚群,结肠管壁内也分布有ICC-IM、ICC-MY和黏膜下ICCs(ICC-SM)3个亚群。结论 沙鼠可用于有关ICCs正常形态、结构及功能的研究。     

全层铺片技术显示胃肠道不同部位Cajal间质细胞

正Cajal间质细胞(interstitial cells of Cajal,ICC),是西班牙神经解剖学家Santiago Ramony Cajal于1893年首次在胃肠道中发现的一类特殊间质细胞,与胃肠道平滑肌和神经纤维末梢在解剖学上有紧密联系,具有独立功能,既是胃肠起搏细胞,又具有传导神经递质的作用~[1]。ICC广泛存在于哺乳动物的整个消化道管壁内,按分布区域可大致分为:分布于胃、小肠和结肠内的环肌与纵行肌之间的肌间神经丛ICC;位于黏膜下环肌层表面的ICC;位于小肠深肌丛区域的ICC;广泛散布于环肌和纵肌肌束内的ICC~[2]。目前消化道全层铺片技术由于能够直观、全面地显示ICC的分布、形态以及形成的细胞网络,而成为研究胃肠神经系统的重要方法之一,但文献中却鲜少提及此方法的详细操作过程。已知ICC特异性表达由原癌基因c-kit编码的受体酪氨酸激酶     

小肠壁内淋巴管的微细分布

用半薄切片光镜观察和超微切片电镜观察，研究了家兔和豚鼠小肠壁内淋巴管的微细分布特点。结果表明：家兔和豚鼠小肠壁内淋巴管起自粘膜层的中央乳糜管，向下注入粘膜层或粘膜下层毛细淋巴管网。在小肠壁的粘膜下层、肌层和浆膜层均有毛细淋巴管和淋巴管。家兔小肠壁淋巴管检出率（７０％）明显高于豚鼠的（４１．３％）。于小肠壁各层内，粘膜层检出率最高（６８．３１％），粘膜下层次之（２５．５７％），肌层较少（５．３３％）     

胎儿十二指肠浆肌层、黏膜下层血管构筑的形态学研究

目的　补充胎儿十二指肠浆肌层及黏膜下层的血管形态学资料。方法　采用组织透明揭层铺片、组织切片及微血管铸型扫描电镜观察三种方法 ,观测 6 0例足月新鲜胎儿十二指肠壁浆肌层及黏膜下层的血管构筑形态。结果　浆膜及纵行肌层毛细血管网主要由直动脉的长支、短支及参与构成黏膜下毛细血管丛第一级血管网的小微动脉发出的返支构成 ;环形肌层毛细血管较稀疏 ,相互连接成长条形血管网 ,毛细血管局部可膨大为窦状 ;黏膜下层毛细血管丛由三级血管网构成。结论　黏膜下层的小动脉为十二指肠器官内血液供应的枢纽     

Morphologic and functional heterogeneous changes of the urinary bladder different parts at rats with infravesical obstruction of the urinary tracts

Infravesical obstruction of the lower urinary tracts of 30 rats was carried out by the measured constriction of the urethral prevesical parts. Morphologic and functional changes of the urinary bladder different parts were studied in 1 week and in 3 months. Compensatory hypertrophy of the detrusor was accompanied increasing of hypertrophied, atrophic and native forms of leiomyocytes, as so their transformation into myofibroblasts with connective tissue formation between the muscle fibers mainly in the neck of urinary bladder Contractility of the detrusor decreased with their tonus increasing mainly in the neck of urinary bladder too. Weakening effect of adrenalin in 3 month after obstruction significantly decreased, while in the body and neck of urinary bladder it virtually disappeared. Blockade of alpha-adrenoreceptors after noradrenalin stimulation resulted in enchancement of the hypertrophic detrusor contraction when tonus of the urinary neck decreased. On the contrary, in the intact urinary bladder the similarly influence resulted in weakening of contraction.     

Sympathetic innervation of the urinary bladder and urethral muscle in the pig

Larsen , J.-J., J. Nordling and B. Christensen , Sympathetic innervation of the urinary bladder and urethral muscle in the pig. Acta physiol. scand. 1978. 104. 485–490. The sympathetic innervation of the lower urinary tract was studied in the female and male pig using a specific histochemical technique for visualization of noradrenaline containing nerves. In smooth muscle from the detrusor few evenly distributed adrenergic nerve terminals were found. A greater number of terminals were observed in the trigone, bladder neck and urethra. There was no sex difference. In smooth muscle specimens from pig detrusor and trigone sympathetic nerve terminals were more abundant than in corresponding tissue from humans.     

构建糖尿病膀胱病豚鼠模型及其尿动力学评价

背景：糖尿病性膀胱病是糖尿病患者最常见的慢性并发症之一,建立糖尿病膀胱病动物模型为相关研究提供实验动物平台。 目的：建立糖尿病膀胱病豚鼠模型并进行尿动力学评价。 方法：50只英国种短毛雌性豚鼠,实验组(n=42)以单次腹腔注射链脲佐菌素法诱导糖尿病豚鼠,对照组(n=8)注射相应剂量空白枸橼酸缓冲液,分别在9周和12周时行尿动力学检查,确定膀胱功能失代偿豚鼠即糖尿病性膀胱病组和代偿豚鼠即代偿组豚鼠尿动力学特点。 结果与结论：42只豚鼠有20只成功诱导出糖尿病。糖尿病豚鼠中,9周时9只糖尿病组豚鼠,6只膀胱功能代偿,3只失代偿;12周时,另外9只糖尿病组豚鼠,1只膀胱功能代偿,8只失代偿(89%)。糖尿病性膀胱病组豚鼠残余尿量增加(0.72±0.08) mL、最大逼尿肌压下降(0.63±0.05) kPa、膀胱容量增加(2.01±0.05) mL及膀胱顺应性增加(3.47±0.41) mL/kPa,与对照组及代偿组比较,差异均存在显著性意义(P < 0.001)。豚鼠一般可在诱导糖尿病成功后12周发生糖尿病性膀胱病,表现出膀胱残余尿量增加等相应的尿动力学改变。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

Heterogeneity of Morphological and Functional Changes in Various Compartments of Rat Urinary Bladder in Infravesical Obstruction of the Urinary Tract

Infravesical obstruction of the lower urinary tract was simulated in rats by dosed constriction of the prevesical portion of the urethra. The functional and morphological changes in various urinary bladder compartments were evaluated after 1 week and 3 months. The development of compensatory hypertrophy of the detrusor was associated with an increase in the number of hypertrophic, atrophic, and young leiomyocyte forms and their transformation into myofibroblasts, with the formation of connective tissue laminae between myofibril bundles mainly in the zone of urinary urinary bladder neck. Specific contractility of the detrusor strips decreased with increasing their tone, which was most pronounced in the neck zone. The relaxing effect of norepinephrine was significantly lower after 3 months of obstruction and virtually disappeared in the zone of the urinary bladder body and neck. Blockade of α-adrenoceptors after adrenostimulation with norepinephrine stimulated contractions of the hypertrophic detrusor against the background of reduced tone of the urinary bladder neck, in contrast to intact urinary bladder where this treatment reduced contractions. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny , Vol. 147, No. 1, pp. 108-112, January, 2009     

Distribution of lumbar and sacral afferent axons in submucosa of cat urinary bladder

Lumbar and sacral afferent axons in the submucosa of the urinary bladder were recognized by degeneration in seven cats subjected to spinal ganglionectomies. Of 2,935 observed terminating axon profiles, 145 were found degenerating. Lumbar afferent axons were 3.7 times more numerous than sacral afferent axons in the submucosa, a reversal of the ratio reported for the muscle coat of the bladder. Sacral afferent axons were evenly distributed to different regions of the bladder, but lumbar afferents were concentrated in the bladder neck. Apparent endings in the submucosa of the urinary bladder were principally free nerve endings. Synaptic vesicles were found in 57% of observed terminating axon profiles. The bladder neck had more terminating axon profiles of all kinds than other regions of the urinary bladder.     

Distribution of lumbar and sacral afferent axons in submucosa of cat urinary bladder.

Lumbar and sacral afferent axons in the submucosa of the urinary bladder were recognized by degeneration in seven cats subjected to spinal ganglionectomies. Of 2,935 observed terminating axon profiles, 145 were found degenerating. Lumber afferent axons were 3.7 times more numerous than sacral afferent axons in the submucosa, a reversal of the ratio reported for the muscle coat of the bladder. Sacral afferent axons were evenly distributed to different regions of the bladder, but lumbar afferents were concentrated in the bladder neck. Apparent afferent endings in the submucosa of the urinary bladder were principally free nerve endings. Synaptic vesicles were found in 57% of observed terminating axon profiles. The bladder neck had more terminating axon profiles of all kinds than other regions of the urinary bladder.     

基于三维重建技术的腰骶椎轴向融合术入路的相关解剖学测量

目的　为腰骶椎轴向融合术入路提供解剖学基础。  方法    利用Mimics10.01对选取的CT数据进行三维重建,以STL格式导入Geomagic studio 12对三维模型进行简单优化,测量相关数据。  结果    以尾骨尖为切口,将轴向融合螺钉固定于腰骶椎体中轴的AxiaLIF手术方式,导针从尾骨尖到钉道入口的距离,男、女性分别为（101.7±9.5）mm,（100.2±9.0）mm（P＞0.05）;以尾骨切迹为切口,将轴向融合螺钉固定在腰骶椎中柱的AxiaLIF手术方式,导针从尾骨切迹到钉道入口的距离,男、女性分别为（82.4±9.5）mm,（83.1±10.6）mm（P＞0.05）;两种不同通路的AxiaLIF手术,导针到S3/4横线的垂直距离EF和EG,男性分别为（24.4±5.3）mm和（14.5±2.8）mm（P<0.05）,女性分别为（27.0±5.6）mm和（17.0±3.8）mm（P<0.05）,有显著的统计学差异。  结论    经尾骨尖为手术切口的AxiaLIF的危险度较高,以尾骨切迹为切口置钉于腰骶椎中柱的手术通路符合骶前间隙安全范围,有待进一步由生物力学研究验证。