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 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 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 analyze the clinical features and genetic characteristics of a patient with Shwachman-Diamond syndrome (SDS) due to compound heterozygous variants of SBDS gene.Methods:A female child with SDS who was admitted to the Children's Hospital Affiliated to Zhengzhou University in February 2022 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her elder sister and parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:The child, a 1-year-and-1-month-old girl, had mainly manifested with diarrhea, hematochezia, growth retardation and malnutrition, along with increased transaminases and decreased neutrophils and hemoglobin. The anteroposterior X-ray of her left wrist indicated significantly delayed bone age. Colonoscopy revealed that her colorectal mucosa was erosive with oily food residues attached to the intestinal lumen. Genetic testing revealed that she has harbored c. 258+ 2T>C and c. 100A>G compound heterozygous variants of the SBDS gene. The c. 258+ 2T>C variant has derived from her father and known to be pathogenic, whilst the other has derived from her mother. Based on the guidelines from the American College of Medical Genetics and Genomics, the c. 100A>G variant was classified as likely pathogenic (PM1+ PM2_Supporting+ PM3+ PM5+ PP3). Conclusion:The compound heterozygous variants of c. 258+ 2T>C and c. 100A>G probably underlay the SDS in this child. For children with refractory diarrhea, liver damage and growth retardation, SDS should be suspected, and genetic testing can facilitate the diagnosis and treatment.

OBJECTIVE: To report on the clinical features and result of genetic testing for a child featuring immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome. METHODS: Clinical records, genetic testing, laboratory investigation and treatment of the child were summarized in addition with a comprehensive review of the literature. RESULTS: The 3-year-old boy was administered due to intractable diarrhea, recurrent infections, liver dysfunction and failure to thrive, though no diabetes or skin disorder was observed. Laboratory testing showed elevated liver enzymes and total IgE, decreased albumin and electrolyte imbalance. Gastrointestinal endoscopy revealed erosion and granules in the duodenum, and edema in the terminal ileum and colon. Biopsies showed villous atrophy in the duodenum and terminal ileum. Genetic testing revealed that the patient has carried a missense c.1087A>G (p.I363V) variant in the exon 10 of the FOXP3 gene. He was treated with enteral and parenteral nutrition, anti infection and Sirolimus, and was waiting for hemopoietic stem cell transplantation. CONCLUSION Although IPEX syndrome usually occur during infancy, it should not be ruled out solely based on the age, and its presentation can be variable. For male children with refractory diarrhea, autoimmune disorder and growth retardation, the diagnosis should be suspected and confirmed by genetic testing.
Child, Preschool ; Diabetes Mellitus, Type 1/genetics* ; Diarrhea/genetics* ; Forkhead Transcription Factors/genetics* ; Genetic Diseases, X-Linked/genetics* ; Genetic Testing ; Humans ; Immune System Diseases/genetics* ; Male ; Mutation ; Polyendocrinopathies, Autoimmune/genetics*

Objective To explore the expression of annexin A10 (ANXA10) in human hepatocellular carcinoma (HCC),and analyze its correlation with matrix metalloproteinases-9 (MMP-9) and vascular endothelial growth factor (VEGF),thus to provide the basis for clinical diagnosis and assessment of HCC.Methods 88 HCC patients were selected,and they were all performed surgical treatment,HCC stage Ⅰ in 11 cases,stage Ⅱ 25 cases,stage Ⅲ 31 cases,stage ⅣV 21 cases.The expressions of ANXA10,MMP-9,VEGF of HCC cancer tissues,adjacent liver tissues and normal liver tissues were tested by immunohistochemical method.The correlation of ANXA10 with VEGF and MMP-9 was analyzed.Results The absorbance value of ANXA10 expression in the HCC cancer tissue was (0.074 ± 0.012),which was lower than that in the adjacent liver tissues [(0.091 ± 0.013)] and normal liver tissues[(0.131 ±0.025)],and ANXA10 expression of the adjacent liver tissue was lower than that of the normal liver tissues,the differences were statistically significant (t =8.96,9.44,8.71,all P ＜ 0.05).The absorbance values of MMP-9 and VEGF expression in the HCC cancer tissue were (0.147 ± 0.017) and (0.127 ± 0.028),respectively,which were higher than those in the adjacent liver tissues [(0.096 ± 0.012),(0.091 ± 0.015)] and normal liver tissues [(0.075 ± 0.014),(0.077 ± 0.019)].The absorbance values of MMP-9 and VEGF of the adjacent liver tissues were higher than those of the normal liver tissues,the differences were statistically significant (t =8.05,9.30,8.11;8.28,9.51,8.02,all P ＜ 0.05).The absorbance value of ANXA 10 expression in the HCC stage Ⅲ + Ⅳ cancer tissue was (0.056 ± 0.010),which was lower than that in the stage Ⅰ + Ⅱ cancer tissue [(0.082 ±0.016)],the difference was statistically significant (t =8.90,P ＜ 0.05).The absorbance values of MMP-9 and VEGF expression in the stage Ⅲ + Ⅳ cancer tissue were (0.157 ± 0.022) and (0.169 ± 0.033),respectively,which were higher than those in the stage Ⅰ + Ⅱ cancer tissue [(0.114 ±0.015),(0.091 ±0.021)],the differences were statistically significant (t =9.13,9.72,all P ＜ 0.05).ANXA10 was correlated with MMP-9 and VEGF (r =0.324,0.295,all P ＜ 0.05).Conclusion ANXA10 presents lower expression in HCC cell,and the expression decreased with the increase of staging.It is negatively related with the MMP-9 and VEGF.ANXA10 expression missing or inactivation of malignant change may be one of the most important features in HCC.

Objective Objective To analysis the clinical curative effect on elderly patients with choledocholithiasis by endoscopic retro-grade cholangiopancreatography ( ERCP) and endoscopic sphincterotomy ( EST) and discuss its clinical significance. Methods 256 patients with choledocholithiasis were collected for diagnosis and treatment by ERCP. The stone characteristics was mastered by ERCP and the pa-tients were treated with EST. Results Patients with choledocholithiasis were successful in 249 cases (97. 27%) for ERCP intubation and 239 cases (93. 36%) for stone remove. After EST,the stones were removed successfully at one time in 199 cases (83. 26%),2 times in 40 cases (16. 74%). 7 patients (2. 73%) with multiple diverticulum and papillary position poor by intubation failure were changed over to op-eration treatment. After ERCP,patients were performed endoscopic nasobiliary drainage (ENBD) 64. 44% (154/239). Pull out the drain-age tube of 90. 91% (140/154) of the patients after stones disappeared which was proved by angiography for 3~5 days. 14 cases (9. 09%) were performed second times to remove the stones due to the presence of residual stones. Postoperative complications occured in 15 cases (6. 02%) including 9 cases of acute pancreatitis and 6 cases of infection of biliary tract, and they were cured after 1 week of corresponding treatment. Transient increase of serum amylase occurred in 39 cases, and all of them recoveried after 3 days without special treatment. Con-clusion The results showed that ERCP ( or EST) were well tolerated by elderly patients with choledocholithiasis,and they were of obvious curative effect and quick recovery,which is worthy of promotion and application.

Objective To explore the effects of zinc finger E-box binding protein (ZEB)2 3′UTR gene transfection on proliferation, invasion and migration in human gastric epithelial cell line GES-1. Methods The synthetic ZEB2 3′UTR and miR-200b micmics were transfected into GES-1 cell line by lipofectamine 2000. We set up control grop, the mutation group and ZEB2 3′UTR group. Real-time quantitative PCR was performed to evaluate the expression levels of miR-200a/b/c and ZEB1/ZEB2 mRNAs after transfection.And then we set up control group, ZEB2 3′UTR group, ZEB2 3′UTR+negative control group and ZEB2 3′UTR+miR-200b micmics group. The protein expression levels of ZEB1, ZEB2, matrix metallopro-teinases (MMP) 2/9 and proliferating cell nuclear antigen (PCNA) were detected by Western blot assay. The invasion and mi-gration capability were analyzed by transwell assay and wound healing test. MTT assay was used to detect the proliferation ability. Results Compared with control group and mutation group, the expressions of miR-200a/b/c were significantly de-creased, especially for miR-200b. And the expressions of ZEB1/ZEB2 were significantly increased at both mRNA and pro-tein levels after transfected with the ZEB2 3′UTR, enhancing the capability of migration,invasion,and proliferation (P <0.05). Compared with ZEB2 3′UTR group, the capabilities of proliferation,invasion and migration were significantly lower in combined group. Conclusion ZEB2 3′UTR can increase the ability of cell proliferation, invasion and metastasis through regulating the levels of miR-200a/b/c, and then influence the regulation of transcription of the target gene, which could lead to malignant transformation of GES-1 cells.

Objective:To explore the effect and mechanism of miRNA sponge on the epithelial-mesenchymal transition (EMT) of gastric carcinoma cell lines SGC7901. Methods:Synthetic ZEB2 3'UTR plasmid and siRNA targeting ZEB2 were transfected into the SGC7901 cell line by Lipofectamine 2000. Real-time quantitative polymerase chain reaction was performed to evaluate the expres-sion levels of miR-200a/b/c. Finally, the migratory, invasive, and proliferative activities of the gastric carcinoma cells in vitro were ana-lyzed by the scratch test, the Transwell cell invasion, and the cell cloning assay. The expression of the target protein was detected by Western blot. Results:Compared with the control group, the expressions of miR-200a/b/c significantly decreased, and their migration, invasion, and proliferation capabilities were considerably higher after they were transfected with ZEB2 3'UTR. Although the expres-sions of miR-200a/b/c significantly increased, the migratory, invasive, and proliferative activities of SGC7901 cells also degraded after they were transfected with siRNA targeting ZEB2. The expression of ZEB2 increased, and that of E-cadherin decreased at the protein level after they were transfected with ZEB2 3'UTR. The protein expression of Vimentin in SGC7901 cells significantly increased. The indicators show the opposite trend when cells were transfected with siZEB2, and the differences between the control and mutation groups were insignificant. Conclusion:ZEB2 3'UTR can regulate EMT course by regulating the miR-200a/b/c expression in gastric car-cinoma, consequently regulating the invasion and migration of carcinoma cells.

Objective To investigate the preventive effect of Indomethacin for post-ERCP pancreatitis and hyperamylasemia.Methods A total of 600 patients,who were undergoing ERCP,were randomly divided into 3 groups to receive anal Indomethacin (n=200),intravenous octreotide (n=200) or no special medication (n=200) before ERCP.The level of serum amylase before and 24h after ERCP were measured,and the rate of acute pancreatitis and hyperamylasemia after ERCP were assessed.Results Serum amylase levels before ERCP of all groups were normal.The mean serum amylase level of Indomethacin group (101.3±77.7 U/L) after ERCP was significantly lower than those of octreotide group ( 176.6±138.3 U/L,P =0.040 ]and control group (227.2±264.9 U/L,P=0.048),while there was no difference between octreotide group and control group ( P＞0.05 ).The incidence of post-ERCP pancreatitis in Indomethacin group (2.5％) was significantly lower than that of control group (9.5％,P=0.003),while there was no difference between octreotide group (4.5％) and control group ( P=0.05 ).The incidence of hyperamylasemia after ERCP in Indomethacin group (5.5％) was significantly lower than that of control group ( 13.5％,P=0.006 ),while there was no difference between octreotide group (10.0％) and control group ( P＞0.05 ).Conctusion Anal administration of Indomethacin before ERCP can effectively reduce the incidence of acute pancreatitis and hyperamylasemia after ERCP.