The Clinicopathologjcal Study And Genetic Diagnosis Of Infantile Cholestasis Diseases

Part One:The clinicopathologjcal study of infantile cholestasis diseasesObjective : Retrospectively control study of the clinical date and liver pathological information of liver biopsy in patients with infantile cholestatic disease, and to analyze its diagnostic significance.Methods : Retrospectively collected the clinical and liver pathological information to classify and summary, who were hospitalized at the Affiliated Hospital, Capital Institute of Pediatrics, between January 2014 to July 2015,referred to Clinical diagnosis of infantile cholestatic disease. According to the results of clinical date and laparoscopic bile duct exploration, patients were divided into non biliary atresia?BA? group and biliary atresia group, and data intercomparison and analysis between the two groups. Inclusion criteria : according to the recommended diagnostic standards of the ninth session of the American Society for Pediatric:ALT and AST increased value,?or? of serum TBil < 85 umol/L?5 mg/d1?, DBIL > 17.1 umol/L?1 mg/dl? or serum TBil > 85 umol/L, DBIL/TBil > 20%, The aged > 1 due to the onset of age in infancy was included in the study.Results:1. There was significantly difference in the level of the onset age, age, hepatomegaly, acholic stool, r- GGT, PT and APTT between non BA group and BA group?P<0.05 or P<0.01?.Whereas CMV infection, splenomegaly, the WBC, HB, ALT, AST, ALP, TBIL, DBIL, and D-Dimer have no obvious difference. 2. Sixty-eight?79.1%? pathological data were collected in our hospital and twenty-six light microscope pathological from 302 hospital. The thrity-seven BA pathology mainly for small bile duct proliferation and hyperplasia of fibre, the maill changes of non BA significant hepatocyte swelling, degeneration and silt, multinucleated giant cell formation and structure of hepatic lobule remaining or not clear. Morphologic changed under light microscope in liver tissues mainly displayed with different levels cholestasis of capillary bile duct and liver cells, liver cell swelling, degeneration, necrosis, multinucleated giant cell formation; small bile duct hyperplasia, atresia and disappeared; expanded portal regions and fibrous tissue hyperplasia, inflammation and fibrosis around hepatic sinus. The most obvious performance was inflammatory cells infiltrating in liver cells and hepatic sinusoids. 3. 68 cases?79.1%? have been confirmed, the liver bile duct system abnormal was accounted for the main reason of cholestasis liver disease, a total of 49 cases?57.0%?. Respectively was biliary atresia?BA? ? 36 cases, biliary atresia?type 1 case, disappear or atresia in interlobular bile ducts was 12 cases(Alagille syndrome?4 cases?. The others were 10 cases of citrin deficiency deficiency, 2 cases of familial intrahepatic cholestasis?PFIC?, 2 cases of congenital high direct bilirubin concentration, 1 case of glycogen storage disease type ?, 2 cases of intravenous nutritional cholestasis liver disease and 2 cases of CMV hepatitis.Conclusion: Contrast the clay stool, r-GGT, hepatomegaly, coagulation function and pathological play an important role in differentiating diagnosis between non BA and BA. BA accounted for a large part of Cholestasis liver disease, the pathological changes were highly specificity than non BA. Inherited metabolic liver disease was higher in non BA group, the diagnosis of it could be made increasingly by combination with liver pathological changes and clinical data. Part Two:Application of captured high throughput sequencing in the genetic diagnosis of infantile cholestasis diseasesObjective:To make genetic diagnosis of infantile cholestasis diseases patients using target gene sequence capture and next generation sequencing technology, aim to discusses the application value of this technology in the genetic metabolic liver disease.Methods:Target gene sequence capture and next generation sequencing were used to 10 patients who were highly suspected genetic metabolic liver disease and hospitalized at the Affiliated Hospital, Capital Institude of Pediatrics, between January 2014 and July 2015. Blood samples were collected from patients and their parents and genomic DNA was extracted from lymphocytes?one patient lack of parents' blood samples?.Sanger sequencing was used to confirm the results of the patients and their parents.Results:4 patients carried 4 pathogenicity mutations in JAG1 gene,including one novel insertion mutation, c.1779-1780 ins A. Three missense mutations, c.703G>A, c.839C>T,c.1720C>T, parental analysis verified that the JAG1 missense mutation of 3 patients were de novo. One patient with glycogen storage disease type ? for compound heterozygous mutations, c.1544 c >T?515 h?, c.1229 a > c. One patient with citrin deficiency for the new frameshift mutations c. 852₈55deltatg and c. 1663₁664inscccgggcagccacctgtaatctc, parental analysis indicated that the mutation respectively from the patient's mother and father. The results of Sanger sequencing validation consistent with the results of the next generation sequencing.Conclusion:Target gene sequence capture and next generation sequencing can detect multiple genes in infantile cholestasis diseases simultaneously,greatly improve the detection efficiency. To provid the molecular diagnosis and treatment for Clinicians have good clinical application prospects.