钙视网膜蛋白在诊断先天性肠无神经节细胞症中的意义

发布时间：2018-10-31 13:28

【摘要】：目的1.探讨钙视网膜蛋白(calretinin,CR)在诊断先天性肠无神经节细胞症(congenital aganglionosis)即先天性巨结肠(hirschsprung's disease,HD)中的概率及意义。2.应用免疫组织化学染色方法,观察对比CR、神经元特异性烯醇酶(neuron-specific enolase,NSE)、B细胞淋巴瘤／白血病-2(B-cell lymphoma/leukemia 2,Bcl-2)在HD病变段及正常肠段肠壁表达情况,评价其在诊断HD中的临床价值。方法1.收集我院2008年1月至2014年10月经病理检查苏木素-伊红(hematoxylin-eosin,HE)染色确诊为HD病例共495例的病案资料,包括钡剂灌肠(contrast enema,CE)、直肠肛管测压(anorectal manometry, ARM)、经直肠粘膜吸引活检(rectal suction biopsy,RSB)等结果,通过上述各项检查阳性率来比较其在诊断HD中的价值,并分析出现CR阴性结果原因。2.收集我院2011年6月至2013年11月巨结肠术后经HE染色确诊为HD的手术标本16例,包括HD病变段及近端切缘(即正常肠段),分别对病变段及正常肠段组织切片进行CR. NSE、Bcl-2免疫组织化学染色,计算机成像系统照相存盘,通过图像分析软件(Image-Pro Plus)判定各标志物在HD病变段及正常肠段神经丛中阳性染色面积百分比。结果1.495例HD患儿中,435例术前CE,265例术前ARM,254例CR检查,其中术前经RSB112例,术后142例。CE、ARM及CR诊断阳性率分别为81.1%、90.6%、99.6%,其中CR阳性率术前为98.2%,术后达100%。2.CR在正常肠壁粘膜下及肌间神经丛呈阳性反应,HD病变段粘膜下及肌间神经丛呈阴性表达。NSE、Bcl-2在正常肠壁粘膜下及肌间神经丛呈阳性反应,HD病变段神经节细胞阴性表达,部分神经纤维阳性表达。3.定量分析：CR染色HID病变段与正常肠段神经丛阳性染色面积百分率存在统计学差异。NSE、 Bcl-2染色HD病变段与正常肠段神经丛阳性染色面积百分率无统计学差异。结论1.诊断HD需依靠病史、典型的临床表现及辅助检查,特殊辅助检查中CR阳性率术前为98.2%,术后达100%。2.CR免疫组化染色可显示正常肠壁神经节细胞结构,呈阳性表达,HD病变段呈阴性表达,CR可作为诊断HD一项重要的神经标志物。3.CR可广泛用于临床,作为术前诊断HD的“金标准”。
[Abstract]:Objective 1. To investigate the probability and significance of calcium retina protein (calretinin,CR) in the diagnosis of (congenital aganglionosis) (congenital megacolon). CR, neuron-specific enolase (neuron-specific enolase,NSE), B cell lymphoma / leukemia-2) was observed by immunohistochemical staining. To evaluate the clinical value of Bcl-2 in the diagnosis of HD. Method 1. From January 2008 to October 2014, we collected 495 cases of HD diagnosed by hematoxylin eosin (hematoxylin-eosin,HE) staining, including barium enema (contrast enema,CE), (anorectal manometry, ARM), of rectal anus manometry. The results of transrectal mucosal aspiration biopsy (rectal suction biopsy,RSB) were used to compare the positive rate of the above examinations in the diagnosis of HD, and to analyze the causes of CR negative results. 2. From June 2011 to November 2013, 16 cases of Hirschsprung's disease diagnosed as HD by HE staining were collected, including the lesion segment of HD and the proximal incisal margin (that is, the normal intestinal segment). The sections of the pathological segment and the normal segment of the intestine were sections of CR. respectively. NSE,Bcl-2 immunohistochemical staining and computer imaging system were used to determine the percentage of positive staining area in HD lesion segment and normal intestinal segment by image analysis software (Image-Pro Plus). Results among the 1.495 cases of HD, 435 cases of preoperative CE,265 were examined by ARM,254 before operation CR, including RSB112 before operation and 142 cases after operation. The positive rates of CE,ARM and CR were 81.1% and 90.6%, 99.6%, respectively. The positive rate of CR was 98.2% before operation. The positive rate of 100%.2.CR was positive in submucous and myenteric plexus of normal intestinal wall after operation. The expression of NSE, was negative in submucous and myenteric plexus of HD lesions. The expression of Bcl-2 was positive in submucous and myenteric plexus of normal intestinal wall, negative in ganglion cells of HD lesion, and positive in some nerve fibers. Quantitative analysis: there was statistical difference in the percentage of positive staining area between the lesion segment of HID and the normal segment of intestine by CR staining, but there was no significant difference in the percentage of positive staining area between the lesion segment of HD and the normal segment of intestine by NSE, Bcl-2 staining. Conclusion 1. The diagnosis of HD depends on the history, typical clinical manifestation and auxiliary examination. The positive rate of CR in special auxiliary examination is 98.2 before operation. The 100%.2.CR immunohistochemical staining after operation can show the normal ganglion cell structure and positive expression. CR can be used as an important neural marker in the diagnosis of HD, and 3.CR can be widely used in clinical practice as the "gold standard" for the diagnosis of HD before operation.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R726.5

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