Progressive family intrahepatic cholestasis （PFIC） is a rare group of autosomal recessive disorders. In recent years， with the development of molecular biology， new pathogenic genes have been constantly identified， and PFIC is currently categorized into 12 genotypes based on the OMIM database. The main manifestations of PFIC include jaundice， pruritus， growth retardation， and malabsorption of fat-soluble vitamins， and some variants can rapidly progress to liver fibrosis， liver cirrhosis， liver failure， and even liver cancer. Different types of PFIC have different clinical manifestations and treatment strategies， and genetic testing can help to achieve early identification and diagnosis. This article reviews the latest advances in the genotyping， clinical features， and treatment of PFIC.

OBJECTIVE: To explore the clinical features and genetic characteristics of three patients with Infantile liver failure syndrome type 2 (ILFS2). METHODS: Three children who were diagnosed with ILFS2 at the Children's Hospital Affiliated to Zhengzhou University from February 2023 to February 2024 were selected as the study subjects. Clinical data of the children were collected. Peripheral blood samples of the children and their parents were collected and subjected to whole exome sequencing (WES). Candidate variants of the NBAS gene were verified by Sanger sequencing. This study was approved by the Ethics Committee of the Children's Hospital Affiliated to Zhengzhou University (Ethics No. 2024-k-069). RESULTS: The three children had presented with fever-triggered recurrent acute liver failure. All of them were found to harbor compound heterozygous variants of the NBAS gene, including c.3596G>A and c.1181A>T in child 1, c.2617C>T and c.2T>C in child 2, and c.3596G>A and c.2817_2818insT in child 3. Among these, the c.1181A>T and c.2817_2818insT variants were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), they were respectively classified as variants of uncertain significance (PM2_Supporting+PM3+PP3) and pathogenic (PVS1+PM2_Supporting+PM3). CONCLUSION Combined with the patient's clinical phenotype, the compound heterozygous variants of the NBAS gene probably underlay the pathogenesis of ILFS2 in the three children. For children with fever-related acute liver failure of unknown causes, the possibility of this disease should be suspected, and genetic testing may facilitate the diagnosis. Early diagnosis and timely intervention can significantly improve the prognosis. Discoveries of the c.1181A>T and c.2817_2818insT variants have enriched the mutational spectrum of the NBAS gene.
Humans ; Exome Sequencing ; Genetic Testing/methods* ; Liver Failure, Acute/etiology* ; Mutation ; Child ; Adult ; Neoplasm Proteins

OBJECTIVE: To explore the clinical and genetic characteristics of a boy with X-linked familial Behcet-like autoinflammatory syndrome-2 (AIFBL2). METHODS: A boy who was admitted to Children's Hospital Affiliated to Zhengzhou University in December 2023 due to recurrent oral ulcers for 2 years, intermittent abdominal pain and fever for more than 1 year was selected as the study subject. Clinical data of the patient was collected. Whole exome sequencing (WES) was carried out, and candidate variant was verified by Sanger sequencing and bioinformatic analysis. A literature search was conducted in OMIM, PubMed, Wanfang Data Knowledge Service Platform, China Biomedical Literature Service System, and the VIP database using the keywords "ELF4 gene" "deficiency in ELF4, X-linked" "ELF4 deficiency" and "DEX" to identify recently published studies. This study was approved by the Medical Ethics Committee of the Children's Hospital Affiliated to Zhengzhou University (Ethics No.: 2023-H-K44). RESULTS: The patient, a 12-year-old male, presented with recurrent mouth ulcers, fever and abdominal pain. Lymphocyte subsets showed a significant decrease in NK cells. Abdominal CT showed thickening of local intestinal wall in the lower right abdomen. Colonoscopy revealed a solitary deep longitudinal ulcer in the ileocecal region. Genetic testing revealed a hemizygote missense variant c.687C>G, with his mother showing the same mutation at this locus. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG), the variant was considered likely pathogenic (PP1+PP2+PM2_Supporting+PP3+PP4). Literature review has found 19 AIFBL2 patients including 1 patient from this study. Mouth ulcer, fever, rash and abdominal pain were the primary clinical manifestations, for which genetic testing is the main diagnostic method. CONCLUSION The hemizygote c.687C>G missense variant of the ELF4 gene probably underlay the AIFBL2 in this child, which has provided a basis for his clinical diagnosis and genetic counseling.
Humans ; Male ; Behcet Syndrome/genetics* ; Child ; DNA-Binding Proteins/genetics* ; Exome Sequencing ; Hereditary Autoinflammatory Diseases/genetics* ; Mutation

Objective:To explore the clinical features and genetic characteristics of three patients with Infantile liver failure syndrome type 2 (ILFS2).Methods:Three children who were diagnosed with ILFS2 at the Children′s Hospital Affiliated to Zhengzhou University from February 2023 to February 2024 were selected as the study subjects. Clinical data of the children were collected. Peripheral blood samples of the children and their parents were collected and subjected to whole exome sequencing (WES). Candidate variants of the NBAS gene were verified by Sanger sequencing. This study was approved by the Ethics Committee of the Children′s Hospital Affiliated to Zhengzhou University (Ethics No. 2024-k-069). Results:The three children had presented with fever-triggered recurrent acute liver failure. All of them were found to harbor compound heterozygous variants of the NBAS gene, including c. 3596G>A and c.1181A>T in child 1, c.2617C>T and c. 2T>C in child 2, and c. 3596G>A and c. 2817_2818insT in child 3. Among these, the c. 1181A>T and c. 2817_2818insT variants were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), they were respectively classified as variants of uncertain significance (PM2_Supporting+ PM3+ PP3) and pathogenic (PVS1+ PM2_Supporting+ PM3). Conclusion:Combined with the patient′s clinical phenotype, the compound heterozygous variants of the NBAS gene probably underlay the pathogenesis of ILFS2 in the three children. For children with fever-related acute liver failure of unknown causes, the possibility of this disease should be suspected, and genetic testing may facilitate the diagnosis. Early diagnosis and timely intervention can significantly improve the prognosis. Discoveries of the c. 1181A>T and c. 2817_2818insT variants have enriched the mutational spectrum of the NBAS gene.

Objective:To explore the clinical and genetic characteristics of a boy with X-linked familial Behcet-like autoinflammatory syndrome-2 (AIFBL2).Methods:A boy who was admitted to Children′s Hospital Affiliated to Zhengzhou University in December 2023 due to recurrent oral ulcers for 2 years, intermittent abdominal pain and fever for more than 1 year was selected as the study subject. Clinical data of the patient was collected. Whole exome sequencing (WES) was carried out, and candidate variant was verified by Sanger sequencing and bioinformatic analysis. A literature search was conducted in OMIM, PubMed, Wanfang Data Knowledge Service Platform, China Biomedical Literature Service System, and the VIP database using the keywords " ELF4 gene" " deficiency in ELF4, X-linked" " ELF4 deficiency" and " DEX" to identify recently published studies. This study was approved by the Medical Ethics Committee of the Children′s Hospital Affiliated to Zhengzhou University (Ethics No.: 2023-H-K44). Results:The patient, a 12-year-old male, presented with recurrent mouth ulcers, fever and abdominal pain. Lymphocyte subsets showed a significant decrease in NK cells. Abdominal CT showed thickening of local intestinal wall in the lower right abdomen. Colonoscopy revealed a solitary deep longitudinal ulcer in the ileocecal region. Genetic testing revealed a hemizygote missense variant c. 687C>G, with his mother showing the same mutation at this locus. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG), the variant was considered likely pathogenic (PP1+ PP2+ PM2_Supporting+ PP3+ PP4). Literature review has found 19 AIFBL2 patients including 1 patient from this study. Mouth ulcer, fever, rash and abdominal pain were the primary clinical manifestations, for which genetic testing is the main diagnostic method.Conclusion:The hemizygote c. 687C>G missense variant of the ELF4 gene probably underlay the AIFBL2 in this child, which has provided a basis for his clinical diagnosis and genetic counseling.

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

Background and aims:Metabolic dysfunction-associated fatty liver disease(MAFLD)is a common chronic condition that can lead to cancer due to its complex pathogenesis.Therapeutic agents targeting AMP-activated protein kinase(AMPK)activation have been suggested as potential treatments for metabolic disorders such as metabolic dysfunction-associated steatohepatitis(MASH).Rhizoma Atractylodis Mac-rocephalae(RAM)has been clinically used to treat obesity-related health problems,but its therapeutic effects on MAFLD and the underlying mechanism remain unclear.Therefore,this study was conducted to evaluate the function and underlying mechanism of RAM in the treatment of MAFLD.Methods:The effect of RAM decoction on MAFLD was evaluated using a high-fat diet(HFD)-induced MAFLD mouse model.In vitro studies were conducted using a palmitic acid/oleic acid-induced lipid accumulation model in the alpha mouse liver 12 cells and RAM-containing serum.The underlying mechanisms were elucidated through a combination of network pharmacology analysis,immunohis-tochemistry,western blotting,and polymerase chain reaction analysis.Results:Administration of RAM decoction significantly reduced body weight gain in MAFLD mice without changing food intake.The weights of the liver and inguinal adipose tissues were also reduced after RAM treatment.Additionally,RAM administration decreased serum levels of alanine aminotrans-ferase,aspartate transaminase,total cholesterol,triglyceride,low-density lipoprotein cholesterol,and glucose,while reducing lipid droplet accumulation in the liver tissues of MAFLD mice.The underlying mechanisms included the activation of the phosphorylation of AMPK and acetyl-CoA carboxylase(ACC),and inhibition of the expression of sterol regulatory element binding protein 1(SREBP1).However,RAM did not alter the protein expression levels of peroxisome proliferator-activated receptor α and carnitine palmitoyltransferase-1α.Furthermore,the RAM-induced upregulation of phosphorylated AMPK,phos-phorylated ACC,and SREBP1 expression,as well as the downregulation of fatty acid synthase expression,were reversed by using an AMPK inhibitor.Conclusions:Through a combination of network pharmacology and experimental validation,we demonstrated that RAM may exert therapeutic effects on MAFLD by inhibiting lipid synthesis and activating phosphorylated AMPK pathways.

Objective To investigate the clinical characteristics of hepatitis-associated aplastic anemia(HAAA)in pediatric patients.Methods A retrospective study was conducted on 212 children with aplastic anemia(AA)who were hospitalized at Henan Children's Hospital from September 2014 to February 2023.The patients were categorized into two groups based on etiology:the HAAA group and the non-HAAA group.The study group consisted of 23 patients in the HAAA group,while a control group of 115 children without HAAA was matched in a 1∶5 ratio based on age,sex,and severity of aplastic anemia.The clinical characteristics,treatment regimens,and outcomes of the 23 patients with HAAA were analyzed and compared with those of the control group comprising 115 patients.Results Among the 23 children with HAAA,there were 11 males and 12 females,with a median age of 6 years and 3 months(ranging from 1 year and 4 months to 12 years old).The onset of aplastic anemia in all HAAA children occurred after the initial presentation of acute hepatitis.Following gradual improvement in liver function,peripheral blood images showed a progressive decline by two or three lines,including platelets.Among these cases,very severe aplastic anemia was observed in 14 patients(60.9%),severe aplastic anemia in 7 patients(30.4%),and non-severe aplastic anemia in 2 patients(8.7%).The median interval between hepatitis onset and diagnosis of aplastic anemia was found to be 56 days(range:10～157 days).All 23 pediatric patients with HAAA presented with acute icteric hepatitis,accounting for 100%of the cohort.One patient(4.3%)was genetically diagnosed with X-linked lymphoproliferative disease type 2,while liver biopsy revealed drug-induced hepatitis/chemical liver injury in five patients(21.7%).Compared to the control group,HAAA patients exhibited significantly lower levels of CD4+cells[(1.2±0.3)vs.(1.5±0.1)]and CD4+/CD8+ratios[(-0.2±0.4)vs.(0.1±0.2)](P<0.05).Three patients received immunosuppressive therapy(IST),18 underwent hematopoietic stem cell transplantation(HSCT),and two non-severe cases were treated with methylprednisolone sodium succinate and compound Zapoan pill;all patients survived.Conclusions Children with HAAA present a critical condition and exhibit a poor prognosis,predominantly manifesting as severe or extremely severe aplastic anemia during the recovery phase of hepatitis.Reduction in CD4+cell count and inversion of CD4+/CD8+ratio may serve as early warning indicators for HAAA.Effective improvement in prognosis can be achieved through immunosuppressive therapy and hematopoietic stem cell transplantation.

Objective:To summarize the clinical phenotype and analyze genetic variant characteristics of hereditary pancreatitis (HP) in children.Methods:Eight families of children diagnosed with HP and admitted to the Children's Hospital Affiliated to Zhengzhou University from August 2022 to May 2024 were selected as the research subjects. A retrospective analysis was conducted on the clinical characteristics, pathogenic gene carrying status, and follow-up of the children. Whole exome sequencing was applied to conduct genetic testing on the 8 children and their relatives. Suspected variations were verified by Sanger sequencing and subjected to bioinformatics analysis.Results:This population was made up of 2 boys and 6 girls with the median onset age of 1.2(0.5-3.4) years. The median age at diagnosis was 2.9(1.5-5.7) years. The clinical symptoms and signs were abdominal pain (8/8), abnormal crying (6/8), vomiting (4/8), and bloating (2/8). None of them had jaundice, fever, hydrothorax or ascites. Pancreatitis generally occurred 4-6 times, and occurred 1-3 times before HP was diagnosed. 6 patients had family history of pancreatitis. Blood amylase and lipase in all 8 patients spiked by over 3 folds. Abdominal imaging showed enlarged pancreas with or without peripheral exudation (8/8), and pancreatic pseudocyst (1/8). The whole exome sequencing detected 2 splicing variant of SPINK1 gene c.194+2T>C(p.IVS3+2T>C), and 6 missense variations of PRSS1 gene c.86A>T(p.N29I) ( n=4) and c.365G>A(p.R122H) ( n=2). Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), all variants were assessed as pathogenic, including 6 inherited from father and 2 from mother. The predicted splicing model indicated that the splice site mutation c.194+2T>C can cause exon skipping, resulting in an unstable truncated protein. Analysis with PyMOL software suggested that the variants c.86A>T (p.N29I) and c.365G>A (p.R122H) may affect the structure of the encoded protein. Conventional conservative measures, including fasting, trypsin secretion inhibition by octreotide, were offered. One child underwent endoscopic retrograde cholangiopancreatogram plus pancreatic duct lithotomy and pancreatic duct stenting. Conclusions:For recurrent childhood pancreatitis, especially with family history, genetic testing is helpful for early diagnosis of HP.

Objective To investigate the clinical characteristics of hepatitis-associated aplastic anemia(HAAA)in pediatric patients.Methods A retrospective study was conducted on 212 children with aplastic anemia(AA)who were hospitalized at Henan Children's Hospital from September 2014 to February 2023.The patients were categorized into two groups based on etiology:the HAAA group and the non-HAAA group.The study group consisted of 23 patients in the HAAA group,while a control group of 115 children without HAAA was matched in a 1∶5 ratio based on age,sex,and severity of aplastic anemia.The clinical characteristics,treatment regimens,and outcomes of the 23 patients with HAAA were analyzed and compared with those of the control group comprising 115 patients.Results Among the 23 children with HAAA,there were 11 males and 12 females,with a median age of 6 years and 3 months(ranging from 1 year and 4 months to 12 years old).The onset of aplastic anemia in all HAAA children occurred after the initial presentation of acute hepatitis.Following gradual improvement in liver function,peripheral blood images showed a progressive decline by two or three lines,including platelets.Among these cases,very severe aplastic anemia was observed in 14 patients(60.9%),severe aplastic anemia in 7 patients(30.4%),and non-severe aplastic anemia in 2 patients(8.7%).The median interval between hepatitis onset and diagnosis of aplastic anemia was found to be 56 days(range:10～157 days).All 23 pediatric patients with HAAA presented with acute icteric hepatitis,accounting for 100%of the cohort.One patient(4.3%)was genetically diagnosed with X-linked lymphoproliferative disease type 2,while liver biopsy revealed drug-induced hepatitis/chemical liver injury in five patients(21.7%).Compared to the control group,HAAA patients exhibited significantly lower levels of CD4+cells[(1.2±0.3)vs.(1.5±0.1)]and CD4+/CD8+ratios[(-0.2±0.4)vs.(0.1±0.2)](P<0.05).Three patients received immunosuppressive therapy(IST),18 underwent hematopoietic stem cell transplantation(HSCT),and two non-severe cases were treated with methylprednisolone sodium succinate and compound Zapoan pill;all patients survived.Conclusions Children with HAAA present a critical condition and exhibit a poor prognosis,predominantly manifesting as severe or extremely severe aplastic anemia during the recovery phase of hepatitis.Reduction in CD4+cell count and inversion of CD4+/CD8+ratio may serve as early warning indicators for HAAA.Effective improvement in prognosis can be achieved through immunosuppressive therapy and hematopoietic stem cell transplantation.