Etiology Analysis Of Pediatric Patients With Hepatopathy:11-year Experience In A Single Center

Objective:To explore the AGL gene mutations in two families affected with glycogen storage disease type Ⅲa.Subjects and methods:The clinical data of the three patients from two families,who were suspected to have glycogen storage disease type Ⅲa,were collected and analyzed.Genomic DNA was extracted from peripheral blood leukocytes of the patients and their parents.All the AGL exons and their flanking sequences of AGL gene were sequenced to identify the causative mutations,and then the frequency of the identified novel viriants was explored by Sanger sequencing analysis of the relevant AGL exons in 75 healthy volunteers.Results:The patient in the first family had bilateral visual loss which was secondary to convulsion caused by hypoglycemia.Laboratory investigations revealed elevated liver and muscular enzymes.The AGL genotype was c.3710₃711delTA/IVS14+1G>T.The former variant was maternaly-inherited and had never been reported,and the latter was an abnormal splice-site variant inherited from the father.The main clinical features of the two siblings in the second family were hypoglycemia and hepatomegaly,along with markedly elevated serum levels of liver and myocardial enzymes.The AGL gene analysis revealed that the patients were both compound heterozygote of the viriants c.1735+1G>T and c.959-1G>C.The splicing viriant c.959-1G>C was a novel one with an allele frequency less than 1%.Conclusions: According to the AGL genetic analytic results as well as their clinical features,the three patients from two families were definitely diagnosed as having glycogen storage disease type Ⅲa and the first one had GSDⅢa in conjunction with retrobulbar neuritis.This research expanded the AGL gene viriant spectrum,provided laboratory evidences for the molecular diagnosis of the patients and for the genetic counseling in the family,and came up with valuable clinical and molecular evidences for the treatment of this disease.Objective: Alagille syndrome（ALGS）is an autosomal dominant disease caused by JAG1 or NOTCH2 variant.This research aimed to explore the clinical and molecular genetic features of ALGS patients.Subjects and methods:The clinical data of 11 patients suspected to have ALGS were collected and analyzed.Genomic DNA was extracted from peripheral blood leukocytes of the patients and their parents.For 5 patients collected before March of 2006,all JAG1 exons and their flanking sequences were directly sequenced.For the remaining 6 patients,high-throughput gene capture technology,chromosomal microarray analysis（CMA）and whole-genome copy-number variant（CNV）analysis when necessary,were utilized to explore the genetic causes.Results: All patients had cholestasis.However,the γ-glutamyl transferase（GGT）levels in one patient were normal.Nine patients had posterior embryotoxon and facial dysmorphism.Eight patients displayed heart anomalies.Seven patients presented with vertebral anomalies and among them,1 patient had sacralization of the cubitus and radius.The condition of nine patients tended to be stabilized on follow-up,but 1 patient died of liver failure in late infancy and 1 got worse.Seven JAG1 variantswere detected in 9 out of the 11 ALGS patients,with c.1977G>A（p.Trp659*）and c.1106₁107del CC（p.Pro369fs）being two novel variants.Two heterozygous interstitial deletions of 3.0Mb and 9.24 Mb in size,respectively,in chromosome 20 were discovered in the remaining 2 patients.Both deletions involved the entire JAG1 gene and the latter had not been reported.De novo origin was unveiled for the detected mutations in most（81.8%）of the patients.Although mother of 2 patients carried the relevant variant,she didn’t demonstrate any clinical features of ALGS.Conclusions: With cholestasis,posterior embryotoxon,facial malformations,heart defects and vertebral anomalies being the major manifestations,ALGS demonstrated variable clinical expressivities and incomplete penetrance.This study identified a total of 7 JAG1 variants as well as 2 interstitial deletions involving this gene,and among them,the variants c.1977G>A（p.Trp659*）and c.1106₁107delCC（p.Pro369fs）as well as the 9.24 Mb chromosomal interstitial deletion had not been reported previously.Objective: This study aims to investigate the etiology distribution feature of pediatrics hepatopathy,so as to provide evidences for their clinical management.Subjects and methods: The research subjects included 590 patients who were diagnosed to have pediatric hepatopathy at the Department of Pediatrics,the First Affiliated Hospital of Jinan University from the beginning of April,2006 to the end of March,2017.By means of a pilot cross-sectional study,their clinical manifestations,laboratory and imaging results,pathology and molecular biology findings were collected and analyzed,and the etiology distribution feature of this cohort was summarized.A chi-square（χ2）test in SPSS version 19.0 was performed to assess the statistical significance of differences in the frequency of pediatrics hepatopathy in Southern and Northern China and age of first diagnosis.Results: Among the 590 patients,there were 357 males（60.5% of the total number）and 233 females（39.5%）.The female-to-male ratio was 1.5 to 1.The medium onset age was one month,with the minimum 1 day and maximum 15 years.There were 291 patients whose onset age was less than one month,244 between 1 month and 1year age,and 55 over 1year age,accounting for 49.3%,41.4% and 9.3% of the total cases,respectively.Among these patients,432 cases（73.2%）had known etiologies,while the etiologies in the rest 158（26.8%）patients remained unclear.In the identified etiologies,genetic hepatopathy,most of which was NICCD（317 cases,53.7%）,took the first place with a proportion of 61.8%（365 cases）of the total patients.Moreover,congenital biliary dysplasia and acquired hepatopathy accounted for 4.6%（27cases）and 6.8%（40 cases）of all the patients,respectively.The patients with congenital biliary dysplasia,acquired hepatopathy as well as NICCD all accounted for larger proportions in South China than in North.Most patients with congenital biliary dysplasia or acquired hepatopathy had their first visit to hospital within the first month of age,while those with NICCD or other genetic hepatopathy,within the age range from 1 month to 1year.Conclusions: By analyzing the clinical and laboratory data of 590 pediatric patients with hepatopathy,the etiology diagnoses were made for 432（73.2%）patients,and 365 patients（61.8%）were found to suffer from genetic hepatopathy,indicating that genetic factors had become the major causes of pediatrics hepatopathy.There were 158 patients with unclear etiologies,the identification of which remained an unresolved issue.Pediatric patients with hepatopathy usually had their first visit to hospital during their infancy,and demonstrated different geographic distribution in terms of their etiologies.This research accumulated scientific data for the etiology spectrum of pediatric hepatopathy,and provided reliable evidences for the diagnosis,treatment and follow-up of the patients with liver diseases.