NFP,TH And Enteric Nervous System Developmental Malformation-HD,HAD

Hirschsprung's disease(HD) was primarily described by Harlald Hirschsprung of Denmark in 1886. HD lies in the second in the inborn diseases of the gastrointestinal tract, about possessing 1/2000-1/5000 of the living infants. The incidence rate of male and female is 3～5:1. HD was occurred 4-12 week of the embryo. The most reason was that the ganglion cell could not locate gather. According to the difference of the intestinal tract that has pathological changes, HD were divided into the short, the long, the whole colon, and the whole digestive tract. The key feature was that the distant intestinal canal was absent or losable of the ganglion cell. The clinical manifestation of Allied Hirschsprung Disorder(HAD) was very similar with HD. But the pathological change was different from HD. Its manifestation were: ganglion cell-penia, ganglion cell-immature, ganglion cell-dystopia, ganglion cell-great. The disease was ignored and incorrectly diagnosed of HD. This disease incidence was about 20%-75% of the HD.Neurofilament protein(NFP), cytoskeletal protein, is the main composition of neurofilaments(NFs). NFs are located specially in the body and prominence of the neuron and the neuroendocrine cell. NFs are divided into light neurofilament(NF-L), mid-size neurofilament(NF-M), and heavy- neurofilament(NF-H) by their molecular mass. Each subunit is the product of single gene. NFs have important roles in the migration,differentiation and development of neuron, evolution of axone and dendron, and transportation of macromolecule.Tyrosine hydroxylase(TH), the marker of sympathetic nerve, is the rate-limiting enzyme of NE. In the wall of intestine there are cholinergic nerve and adrenergic nerve. NE make the smooth muscle of intestine relaxing.The experiment used the immunity-histochemistry. We observed the expression of NFP and TH in the colon of HD,HAD and the normal, in order to invest the role of NFP,TH in neural development. Meanwhile we can comprehend the relationship and distinction between HD and HAD.Our experiment have the HD and HAD models from Tongji Hospital, which were diagnosised as HD and HAD after operation. There were 15 cases of HD, 12 male cases, 3 female cases; there were 11 cases HAD, 8 male cases, 3 female cases. The ages were between 1 month and 9 years. We collected stenotic and dilated segment of bowels from every case. Each paraffin block chiped three slices continuously. The thickness was 6um. At the same time, we selected 10 normal cases as the control. The ages were between 3 years and 8 years.We used the monoclone antibody, adopted S-P immunity-histochemistry. We observed the relationship and difference between the two diseases from two aspects. The first part we compared the expression of NFP+ cell and fiber in the different segment of the three groups. Each group took five slices. We randomly selected five campus visual under the microscope of 200 times from every slice. The images were imported to computer, dealed by cell measurement routine of HMIAS-2000, removal gray scale of bankground. We have statistical analysis to the integral photodensity of the brown-yellow reflexing products, comparing the difference of the three groups in the expression of NFP. The second part we observed the expression of the TH. The method is the same to NFP. Thereby we can offer objective evidence to the diagnosis of HD and HAD.The results of the experiment :①A large number of immunoreactive fibers for NFP and strong staining of the bodies of ganglion cells in the myenteric plexus were observed on the normal specimens and in the dilated segments of colon from HD and HAD(P>0.05). Increasedly immunoreactive fibers for NFP were detected, but there was no NFP-positive cell in the stenotic segment of bowels of HD. On the contrary, NFP-positive cells could be seen in the stenotic segment of the colon from HAD, which were giant cells, small cells and cells with irregular shape, but there was little immunoreactive fibers for NFP. Statistical analysis showed differences in immunoreactivity for NFP between the stenotic segment of colon of HD and the normal control (P<0.01), as well as between HAD and the normal control (P<0.01).②Medium number of immunoreactive fibers for TH and no staining of the bodies of ganglion cells in the myenteric plexuses and submucosal plexuses were observed on the normal specimens. Increasedly immunoreactive fibers for TH were detected in the stenotic segment of bowels of HD. There were little or strong immunoreactive fibers for TH in the stenotic segment of bowels of HAD. And statistical analysis showed significant differences in positive fibers for TH between the stenotic segment of colon of HD and the normal control (P<0.01), moreover statistical analysis showed also difference between HAD and the normal control ((P<0.01)The results of the experiment implied the pathologenesis of HD and HAD was different. The abnormal distribution of NFP and TH could be related to HD and HAD. And the expression of NFP and TH in HD and HAD showed that the pathologenesis of HD and HAD could be different.