The Study On The Assistant Examination And Surgical Procedure For HAD And Immunohistochemical Staining Of Postoperative Paraffin Sections

ObjectiveFirstly, in search of the instructive meaning of the examinational positive index towards diagnose and therapy and the implications of different surgeries on diseases. Generalizing and analyzing of the auxiliary examination (AchE CK-HIS, anorectal manometry) before the surgery and the selections of the operation methods by means of statistics. The selected examples included97Hirschsprung’s disease (HD),28Intestine neuronal dysplasia (IND) and42hypoganglionosis (HG)Secondly, this study selects five markers of neuron:1Protein Gene Product9.5, PGP9.5;2Cathespsin D, CAD;3Peripherin, PR;4S-100;5Neuron-specific enolase, NSE. To study further the characteristic of pathology of Bowel intestinal neuronal dysplasia sticking points and enhance the accurate rate of diagnosis and identification by means of IHC; Immunohistochemistry to examine the expressive character of colon in the conditions of normal、 HD、HG and IND.MethodsTo count the (-) and (+) of the staining of rectal mucosa of the AchE before surgery of the patients in groups HD, IND and HG; adding up no reflective wave, W or U shape wave and sine reflective wave in the anorectal manometry; the surgical methods including leftcolectomy and subtotal coletomy. To color28sections of the paraffin wax of IND colon by means of Immunohistochemical using marked strptavidin/peroxidase among five antibodies: cathepsin D, peripheral protein, S-100and NSE.To observe the size of nerve plexus with optical microscope and the quantities of ganglion cell and so on.ResultsComparative study found that the staining of rectal mucosa of AchE group, showed most patients were negative in group IND, and positive accounted for majority in group HG and HD, this was the unusual difference of IND and HG, HD.As a result of the existence of enteric nervous system, there was no big proliferation of cholinergic neurons in distal colon intestinal wall, this was also the main root and evidence that IND lacking of Hirschsprung spasms of the intestine.Anorectal manometry showed that most of IND patients had rectal-anal inhibition reflex.The most waves were W or U shape, that was different from normal. The operative procedures of IND was subtotal colectomy, oppositely the operative procedures of HG was leftcolectomy, there were significant difference between the HD.The staining characteristic in normal intestinal nervous system were:Catepsins D only was presented in ganglion cell, but was not expressed in nerve plexus; the expression of PGP9.5and Peripherin can be shown in submucosal plexus, myenteric plexus and in the ganglion cells; S-100were expressed in submucosal and myenteric plexus, but expressed negatively within ganglion cells which show a blank area of cell-shape. NSE was brownish strong positive reaction in the cytoplasm of ganglion cells and was yellow fine corrugated expression in circular muscle nerve fibers. But Schwann cells and the surrounding cells were negative.The results of quantitative analysis for HD and AHD were:1Nerve plexus:In IND, the vertical transverse diameter obviously was bigger than that in normal group, the cross-sectional area also obviously was bigger than the normal group, but is smaller than that in HD.2Cells count:the ganglion cells increased markedly in IND, usually6, partially even more than10. ConclusionAchE staining of rectal mueosal biopsy and anorectal manometry are important diagnosis method of HD. They can be used for the diagnosis and differential diagnosis of HG and IND. Surgical operation methods of Hirschsprung’s disease are very more. There are two surgical operation methods of Allied of Hirchsprung’s Disease,we can choose leftcolectomy or subtotal colectomy base on spefical condition. The surgical operation methods of HG are leftcolectomy, however the surgical operation methods of IND are subtotal colectomy.Immunohistochemical staining by using Catepsins D, PGP9.5, S100and Peripherin to examine enteral nervous system have better special result that can obviously show morphological characteristics of the intestinal nerve plexus as well as ganglion cells in IND. This method is helpful for the diagnosis and differential diagnosis in IND.