Genetic analysis of a child with Progressive familial intrahepatic cholestasis type II due to a homozygous variant of ABCB11 gene

To explore the genetic basis for a girl with primary microcephaly and growth retardation. A girl who was admitted to Guangdong Maternal and Child Health Care Hospital in was selected as the study subject. Peripheral blood samples were collected from the child and her parents. Trio whole exome sequencing was carried out, and candidate variants were verified by Sanger sequencing and bioinformatic analysis. This study was approved by the Medical Ethnics Committee of Guangdong Maternal and Child Health Care Hospital (Ethics No. 202201278). DNA sequencing revealed that the child has harbored compound heterozygous variants of the WDR62 gene, including a frameshifting c.2963delC (p.Pro988Argfs*80) variant in exon 24 which was inherited from the unaffected father, and a nonsense c.3163G>T (p.Glu1055*) variant in exon 26, which was inherited from her unaffected mother. Both variants were predicted to affect the reading frame of the WDR62 gene. Based on the clinical manifestations, results of genetic testing and pedigree analysis, the compound heterozygous variants were predicted to underlay the pathogenesis of microcephaly and growth retardation in this child. Above discovery has expanded the mutational spectrum for WDR62-associated Primary microcephaly type 2, and facilitated genetic counseling for the family.

To explore the genetic basis for a woman with oocyte maturation disorder during assisted reproductive treatment (ART), and to verify the source of the variant and its impact on oocyte maturation through family verification. A 35-year-old infertile woman presented at the Reproductive Medicine Center of Gansu Provincial Maternal and Child Health Care Hospital on 20 October 2023 for a 10-year history of infertility despite unprotected intercourse was selected as study subject. Peripheral venous blood sample was collected from the proband. Next-generation sequencing (NGS) was used to detect the potential variant. Candidate variants were validated within her family by Sanger sequencing, and their deleteriousness was assessed with comprehensive bioinformatic analyses to elucidate their origin and impact on oocyte maturation. According to the Standards and Guidelines for the Interpretation of Sequence Variants (hereinafter referred to as ACMG Guidelines) formulated by the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of the candidate variant was rated. This study was approved by the Medical Ethics Committee of Gansu Provincial Maternal and Child Health Care Hospital (Ethics No.: 2023GSFYLS78). The proband underwent three controlled ovarian-stimulation cycles as part of assisted reproductive technology, yielding a total of 29 oocytes, among which only three were mature, whilst the remainders exhibited maturation arrest. Targeted sequencing of peripheral-blood DNA revealed a heterozygous c.728C>T (p.P243L) missense variant of the TUBB8 gene. While the same variant was detected in the proband's father. Based on the ACMG guidelines, the variant was classified to be likely pathogenic (PS4_Supporting+PM2_Supporting+PP2+PP3+PP4). The heterozygous c.728C>T (p.P243L) missense variant of the TUBB8 gene probably underlay the oocyte maturation disorder in the proband, which may be either autosomal dominant or autosomal recessive. For probands with oocyte maturation disorders caused by the heterozygous c.728C>T variant of the TUBB8 gene, oocyte donation may be considered.

The health care department of county-level Maternal and Child Health care Hospital is an important part of maternal and children health care institutions, constructing a set of scientific and standard of performance appraisal index for the health care department is a prerequisite for improving the service objectives of county-level maternal and children health care institutions. This article designed a set of scientific and practical performance evaluation index system for the health care department of county-level Maternal and Child Health care Hospital through literature analysis, in-depth interviews, key performance indicators and Delphy law. The pertinence of evaluation index selection, the distribution rationality of each index weight coefficient and the better operability with scientifically designed evaluation index system could provide a useful reference for the performance appraisal of the health care department of county-level Maternal and Child Health care Hospital.

ObjectiveGeneral hospitals admit lower gestational age neonates than maternal and child health care centers, therefore associated with a higher morbidity and mortality. This study aimed to assess the etiology and clinical characteristics of neonatal sepsis in different medical setting models.MethodsNeonates admitted to 5 tertiary medical centers, including one national general hospital, two maternal and child health care hospitals and two regional general hospitals, in central-south China with culture-proven sepsis between January 2010 and December 2019 were included in the study. We compared maternal and neonatal characteristics, pathogen distribution, treatment and neonatal outcomes among 3 different medical setting models in this retrospective cohort.ResultsWe identified 757 episodes of culture-proven sepsis in 757 neonates. The predominant pathogens were coagulase-negative staphylococci, Klebsiella pneumoniae, Escherichia coli and Group B streptococci. A total of 683 neonates with detailed information were involved in further comparison; 54.6% were from the national general hospital, 35.9% were from the maternal and child health care hospital, and 9.5% were from the regional general hospital. Neonates in national and regional general hospitals had significantly lower gestational age and birthweight (P < 0.001). Patterns of pathogen distribution were different among these medical setting models. Early-onset sepsis was more common in maternal and child health care hospitals (61.4% vs. 42.1% vs. 46.7%, P < 0.001), while hospital-acquired late-onset sepsis was more common in national and regional general hospitals (32.7% vs. 33.3% vs. 11.4%, P < 0.001). The proportion of complications or comorbidities of neonates in maternal and child health care hospitals were significantly lower than neonates in national and regional general hospitals (P < 0.001). The case fatality rate was significantly higher in regional general hospitals (10.8% vs. 3.2% vs. 0.8%, P = 0.001).ConclusionWe report distinct patterns of clinical characteristics, pathogens and outcomes in patient subgroups with neonatal sepsis from national general hospital, maternal and child health care hospital and regional general hospital. It might have some implications for improvement of prevention, management and empirical antibiotic use in neonatal sepsis in different setting models, especially in resource-limited settings from middle and low-income countries.

Introduction: Progressive familial intrahepatic cholestasis (PFIC) type 1 and type 2, characterized by normal serum gamma-glutamyltransferase levels, are caused by defects in FIC1 and BSEP, respectively. Previous reports showed that PFIC type 1 patients had bland intracanalicular cholestasis, whereas non-type 1 patients were marked by non-specific giant cell hepatitis with moderately to markedly elevated serum transaminase levels. However, there has been only one study comparing the clinical features of these two genotypes on the basis of a molecular diagnosis. To investigate the occurrence of genetic mutations and clinical differences between the two genotypes, we performed mutational analysis in FIC1 and BSEP genes in patients with PFIC phenotypes and normal gamma-glutamyltransferase activities. Methods: The coding region and exonintron boundaries of the FIC1 and BSEP gene were sequenced in 17 unrelated Japanese PFIC patients with normal serum gamma-glutamyltransferase activities. All patients were Japanese, from various parts of the country. The following clinical data were collected: age at onset, clinical signs of cholestasis, serum alanine aminotransferase (ALT) levels, liver histology, growth, and therapy including liver transplantation. Results: All patients had novel disease-causing mutations in FIC1 or BSEP. Of the 17 patients, 13 carried FIC1 mutations (5 homozygotes, 7 compound heterozygotes, 1 heterozygote), and 4 carried BSEP mutations (1 homozygote, 2 compound heterozygotes, 1 heterozygote). FIC1 mutations included 8 missense mutations, 2 nonsense mutations, 2 insertions, 1 deletion and 3 splicing mutations. BSEP mutations consisted of 5 missense mutations and 1 exon skipping. There were overlap in ALT levels between PFIC type 1 and PFIC type 2 patients, and other clinicopathological features were insufficient to differentiate between the two genotypes. However, complications such as intractable diarrhea or severe liver steatosis occurred after transplantation in 12 of the 13 patients with type 1, while they never occurred in the type 2 patients. Conclusion: PFIC type 1 is the major type in Japan, and frequently causes severe complications such as intractable diarrhea and liver steatosis after liver transplantation. For predicting these unfavorable complications, genetic diagnosis is needed for case of PFIC before liver transplantation, because it is impossible to distinguish between PFIC type 1 and PFIC type 2 by clinical findings exclusively.

To explore the genetic etiology of a fetus with Coffin-Siris syndrome (CSS). A fetus with abnormal ultrasound findings detected at Luoyang Maternal and Child Health Care Hospital in July 2023 was selected as the study subject. Clinical data were analyzed retrospectively. Whole exome sequencing was carried out on fetal tissue and parental peripheral blood samples, and candidate variant was verified by Sanger sequencing and pathogenicity analysis. This study was approved by the Luoyang Maternal and Child Health Care Hospital (Ethics No. LYFY-YCCZ-2023011). Color Doppler ultrasound at 16+ gestational weeks revealed bilateral ventriculomegaly and cerebellar hypoplasia in the fetus. Trio-WES found that the fetus has harbored a heterozygous c.553C>T (p.Gln185Ter) variant of the ARID1A gene, which was verified by Sanger sequencing to have a de novo origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.553C>T (p.Gln185Ter) variant of the ARID1A gene was classified as pathogenic (PVS1+PS2_Supporting+PM2_Supporting). The fetus was diagnosed with CSS type 2, and the heterozygous c.553C>T (p.Gln185Ter) variant of the ARID1A gene probably underlay its brain malformations.

To explore the clinical manifestations and genetic etiology of a Chinese pedigree affected with Familial focal epilepsy with variable foci (FFEVF). A FFEVF pedigree presented at the Department of Medical Genetics of Linyi Maternal and Child Health Care Hospital on March 14, 2023 was selected as the study subject. The proband was subjected to whole exome sequencing (WES). Candidate variants were verified by Sanger sequencing of the proband and other affected members and bioinformatic analysis. This study was approved by the Linyi Maternal and Child Health Care Hospital (Ethics No. QTL-YXLL-2023048). WES revealed that the proband has harbored a heterozygous c.1642C>T (p.Arg548Cys) missense variant in exon 15 of the NPRL3 gene. Sanger sequencing confirmed that the variant was inherited from the proband's father, and multiple members of the pedigree had also harbored the same variant. Based on guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as variant of unknown significance (PM2_supporting + PP3). The clinical phenotype of FFEVF patients caused by variants of NPRL3 gene is extensive, and the patients may present with neurological abnormality of autism spectrum disorder in addition to seizures.

To explore the clinical phenotype and pathogenesis of a child with partial duplication in the long arm of chromosome 10 (10q), and conduct a review of relevant literature. A child presented at Lianyungang Maternal and Child Health Care Hospital in April 2018 for growth retardation, intellectual disability, and autism spectrum disorder (ASD) was selected as the study subject. Peripheral blood samples were collected from the child and his parents for G-banded chromosomal karyotyping analysis. Genomic DNA was also extracted for chromosomal microarray analysis (CMA). The clinical phenotype and relevant genes were searched in the Online Mendelian Inheritance in Man (OMIM) and the UK Database of Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER). The pathogenicity of chromosomal variation was analyzed based on guidelines from the American College of Medical Genetics and Genomics (ACMG). Relevant literature was searched from the CNKI, Wanfang Data, and PubMed databases by using keywords such as "10q" "duplication" and "trisomy", with the time set as from the establishment of database to December 1, 2023. This study has been approved by the Medical Ethics Committee of the Lianyungang Maternal and Child Health Care Hospital (No. XM2023030). The clinical phenotype of child had included growth retardation, intellectual disability, and ASD. G-banded chromosomal analysis suggested that the child has a karyotype of 46,XY,dup(10)(q23.31q24.33), whilst both of his parents were normal. CMA analysis of the child revealed that the child was arr[19]10q23.31q24.33(87603382_104948862)×3, with a 17.34 Mb duplication in the 10q23.31q24.33 region. Search of the OMIM database suggested that the duplicated segment has contained 171 genes associated with various diseases, and search of the DECIPHER database has identified cases with overlapping with the duplication. A search of the PubMed database has identified 2 publications involving 2 patients with chromosomal duplications overlapping the 10q23.31q24.33 region with a segment length of > 10 Mb. The 2 patients had mainly manifested growth retardation, intellectual disability, ASD, and facial and limb malformations. The main pathogenic genes had included PTEN, WNT8B, LZTS2, NFKB2, PAX2, KIF11, FRA10AC1, and CNNM2. No similar case was retrieved from the CNKI and Wanfang Data databases. The partial 10q duplication as a novel CNV involving genes such as PTEN and WNT8B probably underlay the growth retardation, intellectual disability and ASD in child 1 . This study has enriched the genotype-phenotype spectrum of patients with partial 10q23.31q24.33 duplications.

Progressive familial intrahepatic cholestasis (PFIC) is a rare genetic disorder that results from defective mechanisms of bile secretion. We aim to describe different types of PFIC and their clinical features, treatment modalities, and outcomes in Saudi Arabia. Patients and Methods. This is a retrospective study of all patients diagnosed with PFIC at King Faisal Specialist Hospital and Research Center in Riyadh from January 1, 2002, to December 31, 2021. All relevant information was collected from patient charts and transferred into the REDcap® database for statistical analysis. A total of 79 patients were identified with PFIC, and PFIC type 3 was the most common (59.5%), followed by PFIC type 2 (34.2%), PFIC type 1 (5.1%), and PFIC type 4 (1.3%). Males and females were affected in 54.4% and 45.6%, respectively. Mutations in ATP8B1, ABCB11, and ABCB4 genes were observed in PFIC type 1, PFIC type 2, and PFIC type 3, and loss of function in a variant of TJP2 was detected in PFIC type 4, respectively. A total of 51 (64.6%) patients underwent liver transplantation: three patients (3/4) with PFIC type 1 (75%), twenty patients (20/27) with PFIC type 2 (74.1%), twenty-seven patients (27/47) with PFIC type 3 (57.4%), and one patient with PFIC type 4 (100%). The mean duration of disease before transplantation was 53.9 ± 67 months with a median of 30 months. Following liver transplantation, symptomatic control was achieved in 47 patients (92.2%). Recurrence after transplantation occurred in 4 patients (7.8%) within an average of 22.5 months and a median of 17 months. PFIC is considered a rare disorder in Saudi Arabia; however, early recognition of the disease is important for appropriate management and early referral for liver transplantation evaluation. The overall rate of liver transplantation in our cohort was 64.6% with an excellent five-year survival rate.

To evaluate the impact of Antimicrobial Stewardship Programs (ASPs) on antibiotic use and drug resistance. This was a retrospective, multicenter, management intervention study. The data from 85 maternity hospitals (maternal and child health care hospitals) in Hubei province from 2012 to 2019 were collected. The indicators related to antimicrobial drug use included the utilization rate of different grades of antimicrobial drugs, the intensity of antimicrobial agent use, the rational use of prophylactic antimicrobial agents before class I surgical incision, and pathogenic detection and consultation rates before antimicrobial drug use. Since the implementation, the purchase of antimicrobial agents in hospitals has been maintained within the prescribed range, and the defined daily dose system (DDDs) of antimicrobial agents has been reduced, prophylactic use and accurate treatment of antimicrobial agents related to class I surgical incision have been more reasonable. With the implementation of ASPs, the detection rate of imipenem-resistant Acinetobacter baumannii, cefotaxime-resistant Escherichia coli, and methicillin-resistant Staphylococcus aureus has been decreased in China from national bacterial resistance surveillance data. ASPs have positive effects on antibiotic use and drug resistance in 85 maternity hospitals (maternal and child health care hospitals).

Nasobiliary Drainage in an Episode of Intrahepatic Cholestasis in a Child With Mild <i>ABCB11</i> Disease

To explore the genetic etiology of two fetuses with Mosaic variegated aneuploidy syndrome (MVA) in a pedigree. A 30-year-old pregnant woman, who presented at the Center for Medical Genetics and Prenatal Diagnosis of Shandong Maternal and Child Health Care Hospital on November 16, 2023, was enrolled. Clinical data of the pedigree were collected, and peripheral blood samples from the parents and amniotic fluid samples from the two fetuses were obtained for genomic DNA extraction. Whole exome sequencing (WES) was performed on both fetuses, followed by Sanger sequencing for familial validation and pathogenicity analysis of candidate variants. Chromosomal karyotyping of the parents was conducted to quantify the proportion of premature chromatid separation (PCS). This study was approved by the Medical Ethics Committee of Shandong Maternal and Child Health Care Hospital (Ethics No. 2024-034). Both fetuses exhibited structural brain anomalies and developmental delays during the second trimester. Amniocyte karyotyping revealed low-level mosaic aneuploidy involving multiple chromosomes, while chromosomal microarray analysis (CMA) showed no abnormalities. Pregnancy termination was performed for fetus 1. WES identified compound heterozygous variants in BUB1B, i.e., c.2363_2364del (p.S788Cfs*29) and ss804270619: G>A, in both fetuses. Sanger sequencing confirmed paternal inheritance of c.2363_2364del and maternal inheritance of ss804270619:G>A. According to the American College of Medical Genetics and Genomics (ACMG) and Clinical Genome Resource (ClinGen) Standards and Guidelines for the Interpretation of Sequence Variants, the c.2363_2364del variant was classified as likely pathogenic (PVS1 + PM2_Supporting). Parental karyotyping demonstrated PCS traits, with a higher proportion of abnormal metaphases in the father. The compound heterozygous variants c.2363_2364del (p.S788Cfs*29) and ss804270619: G>A in BUB1B may constitute the genetic etiology of the two MVA fetuses in this pedigree.

Objective To discuss the establishment and application effects of secondary triage management system based on JCI standard in outpatient clinic of maternal and child health care hospital. Methods In July 2015, secondary triage mode was implemented and secondary triage management team was established in our hospital under the JCI standard basis. Nursing staff were trained. The secondary triage management system in outpatient clinic, job and responsibilities of nurses and work flow of secondary triage mode were developed. Satisfaction of patients and doctors in outpatient clinic and waiting time of patients before (January 2014 to June 2015) and after (July 2015 to October 2016) the implementation of secondary triage mode were compared. Results After the implementation of secondary triage mode, the satisfaction of patients increased from (85.76±15.24) to (96.38±12.91) , while satisfaction of patients increased from 88.80% to 97.20%. The waiting time of patients decreased from (59.64±12.38) min to (40.55±10.96) min. The differences were statistically significant (t/χ2=15.634, 13.550, 9.915; P<0.001) . Conclusions The secondary triage management model can reduce the waiting time of patients and improve their satisfaction. Key words: Ambulatory care; JCI standard; Maternal and child health care hospital; Secondary triage; Management

Introduction: Progressive familial intrahepatic cholestasis (PFIC) is a rare group of heterogenous autosomal recessive disorders characterized by defective bile acid secretion or transportation leading to progressive cholestasis. The different types of PFIC are associated with varying genetic defects leading to different phenotypes and extrahepatic manifestations. We report our experience with liver transplantation (LT) in eighteen PFIC patients. Methods: We performed a retrospective review of all children less than 18 years old who underwent LT with a diagnosis of PFIC at our institution. Genetic mutation analysis and explant histopathology were used to confirm all diagnoses. Patient demographics, clinical and laboratory studies, and perioperative data including impact of LT on growth were analyzed. Results: Eighteen children with PFIC underwent LT between November 2005 and October 2018. (Table 1) Subtype analysis based on genetic testing and pathology included two PFIC type 1 (PFIC1), ten PFIC type 2 (PFIC2), two PFIC type 3 (PFIC3), two PFIC type 4 (PFIC4), and two PFIC type 5 (PFIC5). Median age at time of liver transplant was one year of age with the youngest transplant performed on a three-month-old and the oldest transplant performed on a 17-year-old. Two patients with PFIC1 had prior internal cholecystojejunocolostomies for biliary diversion. Two patients, both with PFIC2, had prior partial external biliary diversion procedures. Follow-up from time of transplant ranged from 2 to 15 years, with a median of 8 years. There was improvement in growth in terms of height and weight across all PFIC subtypes. Overall mean height z-score improved from -1.89 to -0.84. Mean weight z-score improved from -1.15 to 0.16. Overall median total bilirubin before transplantation was 7.5 mg/dL with normalization to 0.5 mg/dL posttransplant. There have been no graft failures or deaths in our population. Conclusion: LT is a safe and effective long-term treatment for PFIC of all subtypes. It is associated with good outcomes in terms of growth and control of cholestasis as measured in total bilirubin. There is little morbidity in our population with 100% graft and patient survival even after fifteen years of follow-up.

To analyze the clinical data and genetic characteristics of a child with fibrocartilage hyperplasia type 1 (FBCG1). A child who was admitted to Gansu Provincial Maternity and Child Health Care Hospital on January 21, 2021 due to severe pneumonia and suspected congenital genetic metabolic disorder was selected as the study subject. Clinical data of the child was collected, and genomic DNA was extracted from peripheral blood samples from the child and her parents. Whole exome sequencing (WES) was carried out, and candidate variants were verified by Sanger sequencing. The patient, a 1-month-old girl, had presented with facial dysmorphism, abnormal skeletal development, and clubbing of upper and lower limbs. WES revealed that she has harbored compound heterozygous variants c.3358G>A/c.2295+1G>A of the COL11A1 gene, which has been associated with fibrochondrogenesis. Sanger sequencing has verified that the variants have been respectively inherited from her father and mother, both of whom were phenotypically normal. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.3358G>A variant was graded as likely pathogenic (PM1+PM2_Supporting+PM3+PP3), and so was the c.2295+1G>A variant (PVS1＋PM2_Supporting). The compound heterozygous variants c.3358G>A/c.2295+1G>A probably underlay the disease in this child. Above finding has facilitated definite diagnosis, genetic counseling for her family.