Basic And Clinical Research On Cholestatic Liver Disease In Children

Part ?.Estradiol promotes the differentiation of hepatoblasts into intrahepatic biliary epithelial cells and the development of intrahepatic bile ducts by regulating the Notch signal pathwayAims:To study the effects of estradiol on the differentiation of hepatoblasts into intrahepatic biliary epithelial cells and the development of intrahepatic bile ducts by regulating the Notch signal pathway.Methods:(1)DLK1+hepatoblasts were separated from the liver of C57BL/6CrSlc fetal mice by flow cytometry.The blank control group was subjected to no dispose.The experimental groups were intervened for 24 hours by estradiol with different concentrations(lnmol/L,10nmol/L,100nmol/L),estradiol(10nmol/L)+DAPT(40umol/L).Taqman real-time PCR was used to detect the RNA of Notchl,Notch2,Notch3,Notch4,Jaggedl and DLL4.Laser confocal immunofluorescence staining method was used to detect the expression of Notchl,Notch2,Notch3,Notch4,Jagged 1 and DLL4.(2)The hepatoblasts of blank control group were subjected to no dispose.The experimental groups were intervened for 24 hours by estradiol with different concentrations(lnmol/L,10nmol/L,100nmol/L),estradiol(1nmol/L,10nmol/L,100nmol/L)+DAPT(40umol/L).Flow cytometry was used to detect the percentage of SOX9 and OPN co-positive cells.(3)The C57BL/6CrSlc pregnant mice were respectively divided into control group,estradiol(0.6 mg/kg/day by intraperitoneal injections)group,estradiol(0.6 mg/kg/day by intraperitoneal injections)+DAPT(10 mg/kg/day by subcutaneous injections)group and tamoxifen(0.4mg/kg/day by gavage administration)group for in vivo experiment.Liver samples of the fetal mice at ED 14,ED 18 and newborn mice were collected.Taqman real-time PCR was used to detect the expression of RNA of Notch1,Notch2 and Jagged1.Immunohistochemical analysis was used to detect the expression of Notchl,Notch2 and Jaggedl in the liver.The intrahepatic bile ducts were observed through HE-staining.Results:(1)Estradiol enhances the expression of RNA of Notch1,Notch2,Notch3,Notch4,Jagged1 and DLL4 dose dependently.Estradiol enhances the expression of Notch1,Notch2,Notch3,Notch4,Jagged1 and DLL4.(2)Estradiol promotes the differentiation of hepatoblast into intrahepatic biliary epithelial cells.(3)In vivo,it was proved that estradiol could accelerate the development of bile ducts by activating Notch signal pathway.DAPT can reverse the activation.Tamoxifen can inhibits the development of intrahepatic bile ducts.Conclusions:Estradiol promotes the differentiation of hepatoblasts into intrahepatic biliary epithelial cells and the development of intrahepatic bile ducts by activating the Notch signal pathway.Part ?.Clinical and Histopathologic Features of Sodium Taurocholate Cotransporting Polypeptide Deficiency in Pediatric PatientsAims:The function of NTCP(Sodium taurocholate cotransporting polypeptide)has been the hotspot and focus in recent years.However,little has been reported about the clinical,laboratory and pathological presentations of pediatric patients with NTCP deficiency.The aim of the study was to analyze the clinical,histopathologic and ultra-structural pathologic characteristics of NTCP deficiency in 13 pediatric patients.Methods:From August 2012 to October 2018,this retrospective study conducted in the Department of Pediatrics of Tongji Hospital,China analyzed the data of 13 NTCP deficient patients with an SLC10A1 gene mutation.Except for NTCP deficiency,no other liver diseases were present in the patients,which was determined by both a genetic testing panel for jaundice and by reviewing medical records.The laboratory results,imaging,histopathologic and ultra-structural pathologic information were recorded for analysis.Results:The serum level of total bile acid was high in all 13 patients.All patients had adequate growth and development.Eight of the patients(8/13)presented with visible jaundice and twelve(12/13)were found to have hyperbilirubinemia.A needle liver biopsy was performed in 11 cases,which revealed slightly chronic inflammation in all 11 patients.One of the patients(1/13)was found to be suffering from gallstones.Conclusion:(1)SLC10A1 gene should be detected on patients with hyperbiliary academia especially found through medical examinations.(2)Though most of the patients are asymptomatic or mildly symptomatic,SLC10A1 gene should be detected on patients with cholestasis.(3)SLC10A1 gene should be detected on patients with gallstones of unknown cause.Part ?.Clinical assessment of differential diagnostic methods in infants with cholestasis due to biliary atresia and non-biliary atresia-related cholestasisAims:The differential diagnosis of biliary atresia(BA)and non-BA-related cholestasis was evaluated in order to provide a practical basis for a rapid,early and accurate diagnosis.Methods:396 infants with cholestatic jaundice were studied prospectively during the period of May 2007 to June 2011.The liver function in all subjects was tested.All cases underwent abdominal ultrasonography and duodenal fluid examination.Most cases were subjected to hepatobiliary scintigraphy,magnetic resonance cholangiopancreatography(MRCP)and a percutaneous liver biopsy.The diagnosis of BA was finally made by cholangiography or histopathologic examination.The accuracy,sensitivity,specificity and predictive values of these various methods were compared.Results:178 patients(108 males and 70 females with a mean age of 58±30 days)were diagnosed as having BA.218 patients(136 males and 82 females with a mean age of 61±24 days)were diagnosed as having non-BA etiologies of cholestasis jaundice during the follow-up period in which jaundice faded after treatment with medical therapy.For diagnosis of BA,clinical evaluation,hepatomegaly,stool color,serum gamma-glutamyltranspeptidase(GGT),duodenal juice color,bile acid in duodenal juice,ultrasonography(gallbladder),ultrasonography(griangular cord or strip-apparent hyperechoic foci),hepatobiliary scintigraphy,MRCP,liver biopsy had an accuracy of 76.0%,51.8%,84.3%,70.0%,92.4%,98.0%,90.4%,67.2%,85.3%,83.2%and 96.6%,a sensitivity of 83.1%,87.6%,96.1%,73.7%,90.4%,100%,92.7%,27.5%,100%,89.0%and 97.4%,a specificity of 70.2%,77.5%,74.8%,67.0%,94.0%,96.3%,88.5%,99.5%,73.3%,75.4%and 94.3%,a positive predictive value of 69.0%,72.6%,75.7%,64.6%,92.5%,95.7%,86.8%,98.0%,75.4%,82.6%and 98.0%,and a negative predictive value of 83.6%,8.5%,95.%,75.7%,92.3%,100%,84.2%,93.7%,100%,84.0%and 92.6%,respectively.Conclusions:All the differential diagnosis methods are useful.The test for duodenal drainage and elements is fast and accurate.It is helpful in the differential diagnosis of BA and non-BA etiologies of cholestasis.It shows good practical value clinically.