The Significance Of Calretinin For The Clinical Diagnosis Of Hirschsprung’s Disease

Objective 1.To research the probability and significance of calretinin(CR) for the clinical diagnosis of Hirschsprung’s disease(HD). 2.To observe and compare the distribution of CR, neuron-specific enolase(NSE), B-cell lymphoma/leukemia-2(Bcl-2) by applying immunohistochemical staining in the aganglionic bowel segment of HD and ganglionic bowel segment in order to evaluate the clinical diagnostic value in HD.Method 1.Collected the clinical data of 495 HD cases via pathological examination hematoxylin-eosin(H&E) staining from January 2008 to August 2014, including the outcomes of contrast enema, anorectal manometry and rectal suction biopsy, to compare the clinical value of the examinations by positive rates, and analysis the cause of the negative results of CR.2.Collected the 16 surgical specimens of HD cases via pathological examination H&E staining after operation from June 2011 to November 2013, including the aganglionic bowel segments and ganglionic bowel segments. All specimens were underwent the immunohistochemical staining with CR, NSE, Bcl-2. Performed the immunostaining morphometric analysis by image analysis program(Image-Pro Plus). Evaluated the positive stained area density by calculating the percentage of the area occupied by the positively stained area of the nerve plexus.Result l.In 495 Hirschsprung’s disease children, there were 435 preoperative contrast enema,265 preoperative anorectal manometry,254 CR staining examination which contains 112 by preoperative rectal suction biopsy and 142 post operation. The positive rates of contrast enema, anorectal manometry, rectal suction biopsy were 81.1%、90.6%、99.6%, the positive rate of CR was 98.2% preoperation,100% postoperation.2.CR immunohistochemical staining was positive in submucosal and myenteric plexus in ganglionic bowel segments, negative in aganglionic bowel segments. NSE and Bcl-2 immunohistochemical staining equally presented positive. Both of NSE and Bcl-2 were negative in staining ganglionic cells of aganglionic bowel segments, several nerve fibers were positive.3.The quantitive analysis revealed that there were significant statistical differences in the positively stained area of CR between the ganglionic and aganglionic bowel segments. There were not significant statistical differences in the positively stained area of NSE and Bcl-2 between the ganglionic and aganglionic bowel segments.Conclusion 1.The clinical diagnosis of Hirschsprung’s disease based on medical history, typical physical examination and auxiliary examination. The positive rate of CR was 98.2% preoperation,100% postoperation. 2.CR immunohistochemical staining could show the structure of ganglionic cells, expressed positive of ganglionic bowel segments and negative of aganglionic ones. CR could be an important neural marker in the diagnosis of HD.3.CR could be widely used in clinic, and it could be the ’gold standard’ in preoperative diagnosis of Hirschsprung’s disease.