| Objective. To observe the expression of HuD protein and P2Y2 receptor in the rectal end and normal colon of the middle and high ARM children, and to explore its role on the development of intestinal nervous system in children with ARM.Methods. Terminal rectum specimens of 15 children with middle and high ARM in Zunyi medical college affiliated hospital from July 2014 to January 2016 were collected, According to the position of malformatiorms, there were 15 cases of high and intermediate. The children’s age is 2 days to 5months. Among them, one-stage abdominal-perineal anoplasty were 8 cases, laparoscopic-assisted anorectal pull-through(LAARP) were 7 cases, 10 cases of normal bowel(normal group), 10 specimens and necrosis of intestinal intussusception second phase closed fistula resection of the transverse were 4 cases, high imperforate anus with two closed fistula resection of distal fistula of intestine(co were 3 cases, congenital megacolon secondary closure fistula excision of distal end of the fistulation tube were 3 cases.We collected the distal rectum during operation Sappanwood / eosin staining was used to analyze the number of terminal rectum mucosa and myenteric nerve plexus.Immunohistochemistry(IHC),immunofluorescence and Western blot were used to detect the expression of HUD protein and P2Y2 receptor. A follow-up was conducted on the 15 children from four months to one year, with an average time of five months,and Kelly score was used to assess the anal function.Results. 1.HE staining: The number ofdistal rectum submucous and myenteric plexus in patients with low ARM were significantly deceased as compared with normal controls(7.30±1.26 vs 1.20±0.73, P<0.01).2.IHC: P2Y2 receptor and HuD protein were mainly expressed in the myenteric and the submucosal plexus, predominantly located in the cytoplasm, and HuD protein were also expressed in the nucleus. The expression of P2Y2 receptor in patients with low ARM were significantly reduced with a lower integral optical density as compared with normal controls(18.83±3.65vs8.49±2.68, P<0.01), as well as the expression of HuD protein(43.48±7.11vs3.70±1.21,P<0.01).3. Western blot: The expression of HuD protein in congenital intestinal tissue were significantly lower than normal controls(1.13±0.32vs0.49±0.08, P<0.05), as well as the expression of P2Y2 receptor(0.92±0.18vs0.36±0.09, P<0.05).4.Immunofluorescence staining: The expression of HuD protein were also identified by immunofluorescence staining with a lower integral optical density as compared with normal controls(96.56±17.38vs22.32±4.36, P<0.01), as well as the expression of P2Y2 receptor(78.13±7.86vs26.56±5.32, P<0.01).5. Anal function results:The good rate of anal function in children with ARM was87.7%Conclusion. 1. P2Y2 receptor and HuD protein were expressed in normal colon,mainly in the myenteric and the submucosal plexus, predominantly located in the cytoplasm, while HuD protein were also expressed in the nucleus.2The expression of HuD protein and P2Y2 receptor in ARM children were decreased than normal controls, which may play a role in the development of enteric nervous system. 3 HuD protein and P2Y2 receptor may play a role in the development of enteric nervous system. |
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