Objective Comprehensively analyzing the epilepsy-related genes by bioinformatics methods,and to explore their functions based on network and pathway analysis.Methods I Collected epilepsy-related genes through OMIM,DisGeNET,GeneCards databases and literatures deposited in PUBMED.Performed gene ontology(GO)and pathway enrichment analysis using"ClusterProfiler"R package,and the correlation between pathways was identified through pathway crosstalk analysis.Epilepsy-related genes were then mapped to human protein-protein interaction network(PPIN)to obtain epilepsy-specific PPIN,and extracted the hub and potential genes based on network topology.Results A total of 572 epilepsy-related genes were collected,642 significant GO biological process items and 80 Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways,including learning,memory and cognition,were obtained by enrichment analysis.Based on PPIN analysis,10 hub genes of the epilepsy were extracted and most of them are gamma-aminobutyric acid(GABA)receptor genes.By integrating PPIN and WGCNA analysis,17 potential genes were extracted,involving heat shock protein,growth factor receptor binding protein,etc.Conclusion Epilepsy is a complex process,involving the abnormality of multiple functional genes,multiple biological processes and pathways are closely contact through multiple functional genes,and collectively lead to the occurrence of epilepsy.

Organoids are in vitro 3D tissue cultures that retain the characteristics of their source tissue and organs,and have been widely used in biomedical fields.Organoids have emerged as effective models for investigating diseases,contribute to further understanding of pathophysiology and molecular mechanisms of diseases,and provide technical support for personalized treatment of genetic diseases.This article specifically explores the application of organoid technology in childhood genetic diseases.It emphasizes the advantages and potential advancements of this technology in disease modeling,correction of genetic defects and personalized treatment,thereby serving as a valuable reference for future research endeavors.

Objective: To investigate the pathogenicity of rare damaging variants from ciliary pathway to human lumbosacral neural tube defects(NTDs). Methods: The coding region sequences of 49 ciliary genes were amplified by AmpliSeq technique and sequenced by PGM sequencing platform for screening the rare damaging variants, and the parents without phenotype were used as controls to evaluate the pathogenicity of variants. Results: The rare de novo mutation of GLI3 gene(c.C580 T,p.H194 Y) was detected in one patient.The rare complex heterozygous mutations of CRB2 gene(c.G1392 C,p.R464 S;c.T3448 C,p.C1150 R) was detected in another patient. Conclusion Rare damaging variants from ciliary pathway may be associated with the occurrence of human lumbosacral NTDs.

OBJECTIVE: To carry out genetic testing for two families affected with cobalamin C (cblC) and establish a rapid method for the detection of a hotspot pathogenic variant c.609G>A of the MMACHC gene by using a PCR-high-resolution melting curve (PCR-HRM) method. METHODS: Genomic DNA was extracted from peripheral blood samples of the probands and their parents. Potential variants of the MMACHC gene was analyzed by Sanger sequencing. The c.609G>A variant of the MMACHC gene was screened among 100 healthy children with the PCR-HRM method. RESULTS: Sanger sequencing revealed that proband 1 carried compound heterozygous variants c.394C>T and c.609G>A of the MMACHC gene, while proband 2 carried compound heterozygous variants c.482G>A and c.609G>A of the same gene. PCR-HRM analysis of the two probands and the 100 healthy children were consistent with the Sanger sequencing. CONCLUSION c.609G>A is a hotspot pathogenic variant of the MMACHC gene. The diagnosis of cblC may be rapidly attained through detection by PCR-HRM.

Objective:To investigate the clinical characteristics and treatment of orbital cellulitis in neonates.Methods:Clinical data of 6 neonates with orbital cellulitis were retrospectively analyzed.Newborn orbital cellulitis cases were searched in PubMed, Web of Science, CNKI, Wanfang and VIP databases from the establishment of the database to July 1, 2019, and then relevant literature was reviewed.Results:(1)There were 4 males and 2 females.Five patients had fever.White blood cell(WBC), C-reactive protein(CRP) and procalcitonin(PCT) were significantly increased in 5 cases.Three cases were caused by the spread of infection of surrounding tissues, 2 cases were caused by bacteremia, and 1 case was suspected of vertical transmission from mother to child.Two cases of blood culture and 2 cases of purulent culture were positive.Microbial culture results were all staphylococcus aureus(SA). Four patients were treated with Ceftriaxone + Ampicillin and 2 cases with Ceftriaxone + Linezolid.Abscess incision and drainage were performed in 1 patient.All 6 cases were cured.(2)A total of 13 articles were retrieved, which reported 15 cases of neonatal orbital cellulitis were reported.Together with this 6 cases, 21 cases were analyzed.Periorbital swelling and fever were the main manifestations, and some children presented with simple fever or refuse milk and moan as the first symptom.WBC was significantly increased in most cases.The positive bacterial cultures of 19 cases all contained SA.Nineteen cases were treated with two or more antibiotics.According to the drug sensitivity test results, Vancomycin was used in 8 of the 12 foreign cases, and the third-generation cephalosporin combined with Ampicillin or Linezolid was mainly used in the domestic cases.Abscess incision and drainage were performed in 12 cases.Nineteen cases were cured, 1 case was left with spastic paralysis of limbs, and 1 case died.Conclusions:Orbital cellulitis is rare in the neonatal period, mainly due to the spread of periorbital tissue infection, bacteremia and vertical maternal and infant infection.It is sensitive to oral wiping.Invasive operations increase the risk of infection.The onset of the disease is insidious and ra-pid, and the diagnosis may be delayed in the neonatal period due to the lack of typical symptoms.SA is the chief pathogen.Antibiotics including staphylococci and streptococci should be used for treatment, and drug combination is often required.Incision and drainage should be performed timely for abscess formation.Early diagnosis, rational use of antibio-tics and prompt surgical treatment are necessary conditions for cure.Most of the patients have good prognosis, while the patients with cavernous sinus thrombosis have poor prognosis.

Objective To investigate the spectrum of CYP21A2 gene mutation and the correlation between genotype and phenotype in patients with 21-hydroxylase deficiency in Tianjin and surrounding areas.Methods Genomic DNA was extracted from the peripheral blood samples of the proband.Locus-specific PCR,direct sequencing of PCR amplification products,and multiplex ligation-dependent probe amplification were applied to detect pathogenic gene CYP21A2 and the relationship between genotypes and phenotypes was analyzed.Results (1) Of 35 patients with 21-hydroxylase deficiency,25 were classified as salt-wasting phenotype and 10 were simple virilizing phenotype.(2) 69 mutant alleles were detected in a total of 70 alleles in 35 patients.Only one mutant allele was detected in one patient.Two mutant alleles were detected in all other patients,with the mutation detection rate 98.6％.(3) A total of 6 types of mutations were detected,of which c.293-13C/A＞G (I2G) was the most common,accounting for 57.1％ (40/70),followed by 18.6％ (13/70) for large gene deletion or conversion,and 14.3％ (10/70) for p.I173N.In addition,a novel mutation,c.949C＞T (p.R317X),which has not been reported previously,was detected as a pathogenic mutation.(4) Correlation analysis of genotype and phenotype in 35 children showed that the phenotype predicted by genotype was consistent with the actual salt-wasting phenotype in 31 children,and those in three children were inconsistent with the actual clinical phenotype.Conclusion The mutation characteristics of CYP21A2 gene in patients with 21-hydroxylase deficiency in Tianjin and surrounding areas are slightly different from those reported in other regions in China.A mutation c.949C＞T has not been reported,which enriches the mutation spectrum of CYP21A2 gene and provide the foundation for genetic counseling and prenatal diagnosis.

OBJECTIVE: To detect pathogenic variation in a pedigree affected with hereditary spastic paraplegia type 31 and explore its molecular pathogenesis. METHODS: Customized Roche NimbleGen capture probes were used to capture all exons of the target genes in relation with hereditary spastic paraplegia. The DNA samples were also assayed with fluorescent quantitative PCR as well as chromosomal microarray analysis using CytoScan HD chip. RESULTS: The proband and her father and grandfather were found to carry a deletion for position 85 992 693-86 842 693 on chromosome 2, which spanned approximately 900 kb and encompassed the REEP1 gene. The latter has been specifically associated with hereditary spastic paraplegia type 31. The same deletion was not found in her mother who is phenotypically normal. CONCLUSION The deletional variation of the REEP1 gene probably underlies the disease in this pedigree.
Female ; Humans ; Membrane Transport Proteins ; supply & distribution ; Paraplegia ; Pedigree ; Sequence Deletion ; Spastic Paraplegia, Hereditary ; genetics

Objective: To detect potential mutation in a Chinese pedigree affected with congenital limb malformations. Methods: Clinical data was collected. Genomic DNA was extracted from peripheral blood samples of family members. The zone of polarizing activity regulatory sequence (ZRS) were amplified by PCR and subjected to direct sequencing. Results: Among the 13 individuals in this pedigree, there were 4 PPD patients, who were characterized by varying degrees of deformity. The female patients suffered triphalangeal thumb and preaxial polydactyly, while the male patients only had preaxial polydactyly. Only one patient had foot involvement. TA heterogeneous mutations was discovered in the ZRS (105C>G) in all patients, the same mutation was not detected in 2 healthy family members. Conclusion The inheritance pattern of PPD was autosomal dominant inheritance. There was a significant variability of symptoms among family patients. The heterozygous mutation of the ZRS (105C>G) probably underlie the disease.

OBJECTIVE: To detect potential mutation in a Chinese pedigree affected with congenital limb malformations. METHODS: Clinical data was collected. Genomic DNA was extracted from peripheral blood samples of family members. The zone of polarizing activity regulatory sequence (ZRS) were amplified by PCR and subjected to direct sequencing. RESULTS: Among the 13 individuals in this pedigree, there were 4 PPD patients, who were characterized by varying degrees of deformity. The female patients suffered triphalangeal thumb and preaxial polydactyly, while the male patients only had preaxial polydactyly. Only one patient had foot involvement. TA heterogeneous mutations was discovered in the ZRS (105C>G) in all patients, the same mutation was not detected in 2 healthy family members. CONCLUSION The inheritance pattern of PPD was autosomal dominant inheritance. There was a significant variability of symptoms among family patients. The heterozygous mutation of the ZRS (105C>G) probably underlie the disease.
Female ; Genetic Testing ; Hand Deformities, Congenital ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Male ; Membrane Proteins ; genetics ; Pedigree ; Polydactyly ; genetics ; Thumb ; pathology

OBJECTIVE: To explore the characteristics of differentially methylated genes and gene ontology associated with neural tube defects (NTDs). METHODS: Twelve subjects from 3 NTDs pedigrees were enrolled. Patients with NTDs have served as the case group, while their family members with normal phenotypes have served as the control group. Genomic DNA was extracted from peripheral venous blood samples of the families and used for DNA methylation analysis. Pairwise comparison was carried out primarily for patient-offspring pairs, and co-segregation of methylation pattern with NTDs was analyzed. Pathway related to differentially methylated genes was predicted with DAVID software. RESULTS: Pairwise comparison indicated that VTRNA2-1 was the only gene in which all CpG sites were methylated. Co-segregation of VTRNA2-1 gene methylation with NTDs was found in all pedigrees. Pathways of hypermethylated genes included plasma membrane component, regulation of cellular protein metabolic process, and regulation of actin cytoskeleton organization, while the pathways of hypomethylated genes have included transcription regulator activity, cell adhesion, and neuronal differentiation. CONCLUSION Methylation of the VTRNA2-1 gene has co-segregated with NTDs in the studied pedigrees. The pathways of differentially methylated genes has involved with mechanism of neural tube development.
CpG Islands ; DNA Methylation ; Gene Ontology ; Humans ; MicroRNAs ; genetics ; Neural Tube Defects ; genetics ; Pedigree