Differential expression of microRNAs profile in the peripheral blood of children with drug-resistant epilepsy / 中华实用儿科临床杂志

Objective:To study the profile of microRNAs (miRNAs) in the peripheral blood of children with drug-resistant epilepsy, and to find diagnostic biomarkers for early identification of drug-resistant epilepsy in children.Methods:Retrospective study.Five peripheral blood samples were collected from children in drug-resistant epilepsy group (group R), drug-responsive epilepsy group (group F) composed of the children with epilepsy in pediatric neurology clinic of the First Affiliated Hospital of Zhengzhou University from January 2018 to June 2019 and healthy control group (group J) composed of healthy children who underwent physical examination in the children′s health care clinic at the same time for analyzing miRNA profiles by high-throughput sequencing.In addition, peripheral blood samples were collected from children in R′ group (5 cases), F′ group (7 cases) and J′ group (6 cases) similarly for validating expression levels of 11 candidate miRNAs by quantificational real-time polymerase chain reaction (qPCR). Receiver operating characteristic curves (ROC) were plotted to analyze the diagnostic potential of 7 targeted miRNAs in distinguishing children with drug-resistant epilepsy from drug-responsive epilepsy.Target genes of the 7 validated miRNAs were predicted using online databases, which were then analyzed by Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO). Kruskal-Wallis rank sum test was used for comparison among the three groups.Results:High-throughput sequencing found that compared with group F, there were 68 differentially expressed miRNAs in group R, involving 22 up-regulated and 46 down-regulated miRNAs.qPCR results showed that, expression trends of 7 miRNAs (let-7f, miR-99a-5p, miR-99b-5p, and miR-125a-5p, miR-125b-5p, miR-142-5p, miR-100) were consistent with high-throughput sequencing results among the 11 selected miRNAs.ROC analysis found that when the cut-off values of miR-99a-5p, miR-99b-5p, miR-125a-5p, miR-125b-5p, miR-142-5p and miR-100 were greater than 0.56, 1.00, 3.17, 2.24, 2.09 and 0.59, respectively, their area under curve (AUC) (≥0.871), sensitivity (≥80.0%) and specificity (≥85.7%) were relatively high, which were expected to be diagnostic marker for drug-resistant epilepsy in children.Among them, the diagnostic potential of miR-125b-5p was the best.Bioinformatics analysis found that miR-125b-5p was enriched in the regulation of hypoxia inducible factor-1 signaling pathway, insulin signaling pathway, pluripotent stem cell signaling pathway, mitogen-activated protein kinase signaling pathway, sphingomyelin signaling pathway, neurotrophic protein signaling pathway and mammalian target of rapamycin signaling pathway.Conclusions:The miRNA profile in the whole blood of children with drug-resistant epilepsy is significantly different from that in children with drug-responsive epilepsy.miR-125b-5p is expected to be a potential biomarker of drug-resistant epilepsy in children.

Clinical and genetic analysis of a child with mental retardation autosomal dominant 7 / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical and genetic characteristics of a child with clinical manifestations of hypoplasia, epilepsy and abnormal face.@*METHODS@#The clinical data of the child were collected. The peripheral blood samples of the patient and his parents were extracted for high-throughput sequencing, and Sanger sequencing verification and bioinformatics analysis were performed to detect suspected pathogenic variants.@*RESULTS@#The clinical manifestations of the child were overall developmental backwardness, seizures, autism, and special facial appearance. High throughput sequencing showed that there was a heterozygous mutation of exon 11: c.1920_c.1927delCCTCTACC (p.Ser641Rfs*31) of the DYRK1A gene. The same variant was found in neither of her parents, suggesting that it has a denovo origin.@*CONCLUSION@#The exon11: c.1920_c.1927delCCTCTACC (p.Ser641Rfs*31) mutation in DYRK1A gene was the genetic etiology of the case, which enriches the pathogenic gene spectrum of DYRK1A and provides the basis for clinical diagnosis and genetic counseling.

Different clinical phenotypes and genetic characteristics of five patients with DiGeorge syndrome / 中华内分泌代谢杂志

Objective:To identify the clinical phenotypes, diagnosis, and treatment of five children with DiGeorge syndrome finally diagnosed by gene, with review of the literature.Methods:The clinical data of five children with DiGeorge syndrome admitted to our hospital were collected and sorted out. Copy number variation sequencing (CNV-seq) based on next generation sequencing (NGS) technology was used to diagnose the genetic etiology of the children. The relationship between phenotypes and genotype among these five children were emphatically compared.Results:The five children collected in this study were all younger than 6 months. The course of the disease was more than 2 months to 1 year. Most of the first symptoms were convulsions and/or repeated infection. All of them had different degrees of growth retardation, with or without special facial features, epilepsy, congenital heart disease, etc. The similar blood ionized calcium levels revealed hypocalcemia, but the frequency and severity of convulsions were different. The copy number variation of chromosome 22q11.21 was detected in all these five children, and the deletion fragment was between 2.56-2.6 Mb, which was mostly coincident with the classical deletion region of DiGeorge syndrome (chr22: 19009792-21452445) recorded in Decipher database. One case was suggested to be a novel mutation, and the rest were of unknown origin.Conclusions:DiGeorge syndrome has great clinical heterogeneity. CNV-seq based on NGS technology is not only conducive to accurate genetic etiological diagnosis, but also helpful for understanding the corresponding relationship between clinical phenotype and genotype of hereditary syndrome, improving clinicians′ understanding and avoiding misdiagnosis.

Clinical characteristics and risk factors of post-traumatic seizures in children / 中华实用儿科临床杂志

Objective To analyze the clinical characteristics and risk factors for post-traumatic seizures (PTS) in children,so as to provide a theoretical evidence for clinicians to prevent PTS.Methods From January 2010 to November 2016,the clinical data and auxiliary examination of 388 post-traumatic patients hospitalized at the First Affiliated Hospital of Zhengzhou University were analyzed retrospectively.According to the presence of epileptic seizure,these patients were divided into PTS group and non post-traumatic seizures (nPTS)group.The risk factors associated PTS were investigated by univariate analysis.Results Among the 388 post-traumatic children,72 cases had seizures,which occurred almost predominantly less than 1 year.Fifty-six point nine percent (41/72 cases) were immediately PTS,and 31.9％ (23/72 cases) were early PTS,and 11.1％ (8/72 cases) were late PTS.Among the seizures types,generalized seizures accounted for 51.4％ (37/72 cases),and tonic-clonic seizures were in common;focal seizures accounted for 36.1％ (26/72 cases);focal combined generalized seizures accounted for 2.8％ (2/72 cases),and the remaining 9.7％ (7/72 cases) were ominous.Electroencephalogram showed the slow wave and spike wave most common.There were significant differences in factors statistically,included age,Glasgow Coma Scale (GCS) score,the severity of traumatic brain injury,cerebral contusion,subdural hematoma,therapy method between the patients with seizures group and the patients without seizures group (Z =4.717,x2 =13.079,17.852,5.664,17.457,5.496,all P ＜ 0.05).In single factor analysis and multi-factor regression analysis,age,GCS score,the severity of traumatic brain injury,subdural hematoma,therapy method were associated with the incidence of PTS (all P ＜ 0.05).Conclusions PTS is a severe complication of brain trauma in children.Small age,GCS ≤8 scores,severe brain injury,subdural hematoma,surgery are the risk factors of PTS.

Identification of pathogenic mutation in a Chinese pedigree affected with split hand/split foot malformation / 中华医学遗传学杂志

OBJECTIVE@#To detect potential mutation in a Chinese pedigree affected with split hand/split foot malformation (SHFM).@*METHODS@#The patients were screened for genome-wide copy number variations with single nucleotide polymorphism (SNP) microarray. Copy number variations were verified by real-time fluorescence quantitative PCR.@*RESULTS@#There were 3 SHFM patients from three generations, which conformed to an autosomal dominant inheritance. SNP microarray assay revealed that all patients have carried a 0.34 Mb duplication in 10q24.31-q24.32 (102 993 649-103 333 271) encompassing the BTRC and DPCD genes. The result was verified by real-time fluorescence quantitative PCR, confirming that the duplication has co-segregated with the SHFM phenotype in the pedigree.@*CONCLUSION@#The 10q24.31-q24.32 duplication probably underlies the pathogenesis of SHFM in this pedigree. Tiny copy number variations can result in diseases featuring autosomal dominant inheritance.

Study on glycyrrhizin in reducing neuronal damage by inhibiting high mobility group protein 1 in immature rats with epilepsy / 中华实用儿科临床杂志

Objective To study the effect of glycyrrhizin(GL) on the gene expression of high mobility group protein 1 (HMGB1) in hippocampus and serum.To evaluate the effect on the expression of neuron-specific nuclear-binding protein (Neu-N) in the hippocampus CA1,CA3 regions in the chronic stage of an immature rat epilepsy model.Methods Fifty-two 21 day-old SD rats were randomly divided into control group,model group Ⅰ and model group Ⅱ according to the random table method.Model group Ⅰ was induced epilepsy by kainic acid (KA),and the model group Ⅱ was pretreated with GL by intraperitoneal injection at 30 min before KA injection.According to the different observation time points,each group was divided into 4 subgroups:3 h,12 h,24 h and 7 d.Model group Ⅱ was divided into 3 subgroups:10 mg/kg,50 mg/kg,100 mg/kg,according to the different doses of GL.There were 3 animals in each subgroup.Score was performed according to the Racine score,and quantitative real-time polymerase chain reaction and Western blot were applied to detect the mRNA and protein expression of HMGB1 in the acute phase.Enzyme-linked immunosorbent assay(ELISA) was applied to measure the expression of HMGB1 in blood;immunohistochemical was applied to measure the expression of Neu-N in hippocampus in the chronic phase(7 d).Results Compared with model group Ⅰ,seizure onset time was obviously prolonged in model group Ⅱ [(24.08 ± 1.98) min vs.(33.39 ± 2.66) min],and the difference was statistically significant (t =9.231,P ＜0.05);Comparing KA model group Ⅰ with control group,the gene expression of HMGB1 significantly increased,and reached a peak at the time of 12 h (H =10.532,P ＜ 0.05),but the protein expression of HMGB1 was changed obviously and there was no significant difference (H =5.227,P ＞0.05).The expression of HMGB1 in the serum also significantly increased,especially at 12 h (H =6.897,P ＜0.05).At the time of 12 h after KA injection,the gene expression of HMGB1 in the hippocampus was significantly decreased in model group Ⅱ compared with model group Ⅰ (H =10.721,P ＜0.05) (especially in the 100 mg/kg model group).Also,the expression of HMGB1 in the scrum was obviously decreased (H =6.967,P ＜ 0.05) (especially in the 100 mg/kg model group).At the time of 7 d after KA injection,hippocampal neuron loss in model group.Ⅰ was significantly reduced compared with control group (P ＜ 0.05),and hippocampal neuron loss in model group Ⅱ was evidently decreased compared with model group Ⅰ (P ＜ 0.05),(especially in the 100 mg/kg model group in CA1,50 mg/kg model group in CA3).Conclusions In the immature rat temporal lobe epilepsy model,GL may have neuroprotective by inhibiting the synthesis and release of HMGB1,inhibiting inflammation further to restrain the loss of neurons in the chronic phase.