The Immunohistochemical Staining in Determination of Postoperative Bowel Dysfunction in Children with Hirschsprung's Disease

Document Type : Original Article (s)

Authors

1 Associate Professor, Department of Surgery, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Resident, Department of Surgery, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Assistant Professor, Department of Pathology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: The aim of this study was to evaluate the calretinin and nitric oxide (NO) immunohistochemical (IHC) staining in ganglionic segment of colon of children with Hirschsprung's disease (HD).Methods: In a cross-sectional study, 60 children with HD entered the study, and underwent surgery to pull out the bowel from the anus to take biopsy from every 5 cm. Hematoxylin and eosin (H&E) and IHC methods were evaluated, and the results were reported to be poorly positive or severely positive for calretinin. All patients were followed up 1 and 6 months after the operation, and the results were compared between the two groups.Findings: From 60 children with HD, 14 patients (23.3%) were symptomatic. In 14 symptomatic patients, all colorectal samples from ganglionic segments were negative for NO staining in comparison of placental NO staining as sham group. In the IHC examination, nuclear and cytoplasmic calretinin staining revealed ganglion cells in 52 patients as strongly positive, and weakly positive in 8 patients. Six patients in strongly positive group (11.5%) were symptomatic, while 8 patients whose calretinin staining was reported as weakly stained, had symptoms (P = 0.001). Overall accuracy of prediction of postoperative constipation for calretinin staining was 96.67%.Conclusion: According to the findings of our study, it seems that in children with HD who undergo surgery, proximal ganglion immunohistochemical staining may be effective in determining postoperative bowel dysfunction.

Keywords


  1. Hiradfar M, Sharifi N, Khajedaluee M, Zabolinejad N, Taraz JS. Calretinin immunohistochemistery: An aid in the diagnosis of Hirschsprung's disease. Iran J Basic Med Sci 2012; 15(5): 1053-9.
  2. Cheng LS, Schwartz DM, Hotta R, Graham HK, Goldstein AM. Bowel dysfunction following pullthrough surgery is associated with an overabundance of nitrergic neurons in Hirschsprung disease. J Pediatr Surg 2016; 51(11): 1834-8.
  3. Parmentier M, Passage E, Vassart G, Mattei MG. The human calbindin D28k (CALB1) and calretinin (CALB2) genes are located at 8q21.3----q22.1 and 16q22----q23, respectively, suggesting a common duplication with the carbonic anhydrase isozyme loci. Cytogenet Cell Genet 1991; 57(1): 41-3.
  4. Alexandrescu S, Rosenberg H, Tatevian N. Role of calretinin immunohistochemical stain in evaluation of Hirschsprung disease: an institutional experience. Int J Clin Exp Pathol 2013; 6(12): 2955-61.
  5. Goldstein AM, Hofstra RM, Burns AJ. Building a brain in the gut: development of the enteric nervous system. Clin Genet 2013; 83(4): 307-16.
  6. Vorobyov GI, Achkasov SI, Biryukov OM. Clinical features' diagnostics and treatment of Hirschsprung's disease in adults. Colorectal Dis 2010; 12(12): 1242-8.
  7. Neuvonen MI, Kyrklund K, Rintala RJ, Pakarinen MP. Bowel function and quality of life after transanal endorectal pull-through for hirschsprung disease: Controlled outcomes up to adulthood. Ann Surg 2017; 265(3): 622-9.
  8. Jarvi K, Laitakari EM, Koivusalo A, Rintala RJ, Pakarinen MP. Bowel function and gastrointestinal quality of life among adults operated for Hirschsprung disease during childhood: A population-based study. Ann Surg 2010; 252(6): 977-81.
  9. Young HM. All is not normal in "Normoganglionic" bowel regions of Hirschsprung disease model mice. Cell Mol Gastroenterol Hepatol 2015; 1(1): 4-5.
  10. Coyle D, O'Donnell AM, Gillick J, Puri P. Altered neurotransmitter expression profile in the ganglionic bowel in Hirschsprung's disease. J Pediatr Surg 2016; 51(5): 762-9.
  11. Wu TT, Tsai TW, Shen YT, Hsu JD, Yang LC, Li C. Analyses of PRMT1 proteins in human colon tissues from Hirschsprung disease patients. Neurogastroenterol Motil 2010; 22(9): 984-90, e254.
  12. Musser MA, Correa H, Southard-Smith EM. Enteric neuron imbalance and proximal dysmotility in ganglionated intestine of the Sox10(Dom/+) Hirschsprung mouse model. Cell Mol Gastroenterol Hepatol 2015; 1(1): 87-101.
  13. Zaitoun I, Erickson CS, Barlow AJ, Klein TR, Heneghan AF, Pierre JF, et al. Altered neuronal density and neurotransmitter expression in the ganglionated region of Ednrb null mice: Implications for Hirschsprung's disease. Neurogastroenterol Motil 2013; 25(3): e233-e244.
  14. Musa ZA, Qasim BJ, Ghazi HF, Al Shaikhly AW. Diagnostic roles of calretinin in hirschsprung disease: A comparison to neuron-specific enolase. Saudi J Gastroenterol 2017; 23(1): 60-6.
  15. Kapur RP, Reed RC, Finn LS, Patterson K, Johanson J, Rutledge JC. Calretinin immunohistochemistry versus acetylcholinesterase histochemistry in the evaluation of suction rectal biopsies for Hirschsprung disease. Pediatr Dev Pathol 2009; 12(1): 6-15.
  16. Kacar A, Arikok AT, Azili MN, Ekberli AG, Tiryaki T. Calretinin immunohistochemistry in Hirschsprung's disease: An adjunct to formalin-based diagnosis. Turk J Gastroenterol 2012; 23(3): 226-33.
  17. Holland SK, Ramalingam P, Podolsky RH, Reid-Nicholson MD, Lee JR. Calretinin immunostaining as an adjunct in the diagnosis of Hirschsprung disease. Ann Diagn Pathol 2011; 15(5): 323-8.