摘要目的 探讨还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)、乙酰胆碱酯酶(AchE)酶组织染色法、快速苏木素-伊红(HE)染色法在先天性巨结肠(HD)术中诊断中的优劣,提高术中诊断准确率,减少再次手术风险.方法 标本取自HD患儿术中切除的结肠18例,其中男11例,女7例,年龄(8.6±3.0)个月.标本分狭窄段、移行段、扩张段,分别用NADPH-d的快速染色法、冰冻切片HE与AchE酶组织染色法快速染色法进行比较,并用β-Ⅲ型微管蛋白(TUJ-1)免疫荧光再次证实NADPH-d的快速染色法的准确性.结果 3种方法诊断HD的阳性率分别是NADPH-d法94.4％ (17/18),HE法83.3％ (15/18),AchE酶组织染色法88.9％(16/18),组间差异无统计学意义(P＞0.05).NADPH-d法耗时最短,为(7.0±1.7)min,HE法(23.2±2.9)min,AchE法(30.9±2.2) min,各组间比较差异有统计学意义(P＜0.01).结论 NADPH-d法和HE法联用能术中快速诊断HD,确定病变段肠管切除范围.AchE酶组织染色法适用于黏膜层的活检诊断.

abstractsObjective To evaluate the superiorities and disadvantages of reduced nicotinamide adenine dinucleotide dehydrogenase (NADPH-d),acetylcholinesterase (AchE) enzyme histochemistry and hematoxylin and eosin (HE) staining in the intraoperative rapid diagnosis of Hirschprung' s disease (HD),increase the intraoperative diagnostic accuracy and decrease the risks of reoperation.Methods Eighteen children with HD underwent intraoperative full-thickness intestinal biopsies,including 11 boys,and 7 girls,with average age of (8.6 ± 3) months.Frozen sections at three levels (narrow segments,transitional region and dilated segments) were taken,and the extent of aganglionosis was examined by using NADPH-d staining,traditional rapid HE and AchE staining respectively.TUJ-1,a marker of enteric neural cells,was used for identification of NADPH-d positive myenteric plexus and ganglion cells by immunofluorescence staining.Results The positve rate of NADPH-d staining,HE staining and AchE staining for HD was 94.4％ (17/18),83.3％ (15/18),and 88.9％ (16/18),respectively (P ＞ 0.05).Time comsuming in NADPH-d,HE and AchE staining was (7.0 ± 1.7),(23.2 ± 2.9) and (30.9 ± 2.2) min,respectively(P ＜ 0.01).Conclusion The combination of NADPH-d and HE staining,with high idiosyncrasy and sensibility,simple and convenient,can be used as a routine intraoperative diagnosis method to identify HD and detect cutting range of colon.AchE staining was the best technique to demonstrate hypertrophic nerve trunks in the mucosa and submucosa and was well suited to suction rectal biopsy.