| Objective: To explore the effect of enteric nervous system(ENS) in pathogenesis of Congenital choledochal cyst (CCC), the expresses of nervous growth factor (NGF) with their acceptors (high affinity receptor TrkA and low affinity receptor P75NGFR ), neurocyte specific enolase(NSE) and the production of soluble protein gene 100 (S-100) in CCC were observed by comparing with above-mentioned ones in the biliary tract in fetus. At the same time, because the detection of NSE and S-100 has a cooperative and complement feature, it is regarded as a fine mark for judging the growth degree of biliary tract neurons in order to fill up the mechanism of CCC further more.Methods: Between January 2007 and January 2009, 22 cases with CCC were enrolled in this study in department of pediatric surgery of the Second Hospital of Hebei Medical University. There were 4 male ones and 18 female ones, whose ages were 4.44±3.90 years(1 month-18 years). Of them, 14 cases had an abdominal pain, 6 cases presented jaudice and white stool, 1 case with fever and vomiting, 1 case was found by ultrasonographic investigation. They were all diagnosed clearly by ultrasonographic investigation or CT in preoperation. We resected their choledochal cysts and gallbladders, to practice hepaticojejunm Roux-Y stomy. We took the wall of gall bladders, the proximal and distal segments of duct cysts as experimental group samples. 11 cases who were termined of pregnacy in fetal phase was control group, there were 5 male ones and 6 female ones, whose moon's age were 7.36±0.81 months(6-8 mongths). After being prepared by means of being fixed by 10% formaldehyde and imbedded by paraffm, paraffm section were examined by immunohistochemistry and restained by hematoxylin in order to make the visualization more clear. The ones whose cytoplasms showed brown color were positive, or else, the ones showed blue color were negative. Observing the paraffm section by optical microscope, we get the message about neural development and distribution in bile duct wall. The positive rate of cells were divided to five grades (A) according to the certerition as follow: 0~1%=0, 2~10%=1, 11~50%=2, 51~80%=3, 81~100%=4. The positive strength of cells were divided four grades(B) by Image-Pro Plus: 0(negative), 1(weak positive), 2(positive), 3(strong positive). Then the quantities of the positive cells (IHS) in five field of visions were caculated (IHS =A×B). All of the data were analyzed with SPSS version 13.0 software for Windows (SPSS, Inc., Chicago, USA). The one-factor analysis varianc test was used to compare the expresses of all indexes in the walls of cysts or gall bladders of CCC and those of fetus. P values <0.05 were considered statistically significant.Results: NGF and its acceptors ( TrkA and P75NGFR ), NSE and S-100 were all expressed at different levels in the cytoplasm of two groups which being observed by light microscope, then the quantities of the positive cells in five fields of visions were caculated by Image-Pro Plus. All of the data were analyzed by SPSS 13.0 for Windows by one-factor analysis varianc test. The expresses of NGF, TrkA, P75NGFR and NSE in distal segments of CCC were 0.73±0.15, 0.82±0.13, 0.91±0.13 and 0.73±0.12 respectively; meanwhile, the indexes above-mentioned expressed in proximal segments of CCC were 1.50±0.18, 2.14±0.34, 1.50±0.25 and 1.50±0.25. Furthermore, they were expressed in bile duct of fetus phase: 2.64±0.49, 3.82±0.50, 2.27±0.52 and 2.36±1.56. The expresses of NGF, TrkA, P75NGFR and NSE in distal segments of CCC were weaker than those in proximal segments of CCC; nevertherless, the significant difference lied in the express rate of the indexes above-mentioned in the distal and proximal of CCC and those in bile duct of fetus phase (P﹤0.05). The expresses of S-100 in distal segments, proximal segments of CCC and the bile duct of fetal phase were 1.00±0.11, 1.59±0.20 and 2.73±0.38. Compared to the express of S-100 in bile duct of fetus phase, the express of S-100 in distal and proximal segments of CCC were all weaker(P﹤0.05), but there was no statistical significance between the express of S-100 in the distal segments and proximal segments of CCC (P﹥0.05). In the end, there was no statistical significant significance about the expresses of all above-mentioned indexes in gall bladder walls between the two groups (P﹥0.05).Conclusions:1. Abnormal development of enteric neurons is possibly one of the pathogenetics of CCC on the basis of which the expresses of NGF, TrkA and P75NGFR were all weaker in CCC than those in bile duct of fetal phase.2. The expresses of NGF, TrkA and P75NGFR in distal segments of CCC were weaker than those in proximal segments of CCC which suggests that the distal segment of CCC is the major pathological point which results in the proximal bile duct expanded gradually.3. The expresses of NGF, TrkA and P75NGFR in proximal segments of CCC were all weaker than those in bile duct of fetal phase. The lower density of obove-mentioned indexes is due to the expandance of the proximal bile duct or the abnormal neural development of the proximal segments of cysts. This conclusion is wait for the further study in the future.4. NGF, TrkA and P75NGFR are possibly important to the pathogenesy of CCC without influence to the development of gall bladder.5. NSE and S-100 are the markers of matural development of biliary tract nerves. Compared to the expresses of them in bile duct of fetal phase, the expresses in CCC were weaker. The result indicate that the development of NGF, TrkA and P75NGFR are all confined in choledochal cyst.
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