Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions: Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.
目的： 研究钙视网膜蛋白(calretinin，CR)在全结肠无神经节细胞症(total colonic aganglionosis, TCA)不同肠段的表达，探讨其临床意义。 方法： 收集华中科技大学同济医学院附属武汉儿童医院2010至2016年TCA行回肠结肠根治手术病例9例，另选取9例因其他疾病切除肠管的正常手术切断缘作为对照。对其进行常规HE及免疫组织化学EnVision法染色，观察CR在结肠、回肠及TCA不同肠段中的表达模式，并对黏膜固有层及黏膜肌层内CR阳性纤维进行计数，同时应用图像分析软件(Image-Pro-Plus)判定CR阳性染色在黏膜固有膜内的平均吸光度，分析其在不同肠段中的表达差异。 结果： CR在回肠和结肠中的染色模式相似，含神经节细胞的回肠和结肠黏膜固有膜和黏膜肌层均可见CR染色阳性纤维，其数量差异无统计学意义(P>0.05)；无节细胞的回肠和结肠CR阳性纤维数量显著少于对照组和切断缘(P<0.01)；回肠移行段黏膜CR阳性神经纤维数量明显少于回肠切断缘(P<0.01)，其黏膜下及肌间可见小的神经丛，CR神经纤维呈不连续的束状染色。 结论： CR在TCA含节细胞肠段和无节细胞肠段的表达存在显著差异，具备辅助诊断TCA的潜在价值，有望成为诊断TCA的重要方法。.
Keywords: Calretinin; Hirschsprung disease; Immunohistochemistry; Intestinal diseases.