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.

Case report of 16p11.2 microdeletion only manifested as paroxysmal kinesigenic dyskinesia / 中华实用儿科临床杂志

The clinical data of a child with paroxysmal kinesigenic dyskinesia (PKD) and being diagnosed and treated in the Department of Pediatrics of the First Affiliated Hospital of Zhengzhou University in October 2018 were analyzed retrospectively.The male patient was 13 years old.The clinical manifestation was the change of body position, and the temporary movement cannot appear.The manifestations included the turning of head to one side, the falling back of neck, head shaking, swinging, the tightly hugging of hands in front of the chest, the touching of two tiptoes to the ground, numb sole, and ache.Gene detection: chromosome 16p11.2 (chr16: 29594293-30189789) had about 595.5 kb heterozygosity deletion.A total of 8 cases of 16p11.2 microdeletion in children with PKD were reported in details.16p11.2 microdeletion is another form of gene expression that causes PKD.16p11.2 microdeletion should be screened for genetic evaluation in patients with PKD.

Case report of anti-AMPA2 receptor antibody encephalitis after herpes simplex virus encephalitis in child / 中华实用儿科临床杂志

The clinical data of a child with anti-(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid, AMPA2) receptor antibody encephalitis after herpes simplex encephalitis was retrospectively analyzed.The child was a 9-year-old female developing abnormal mental behavior after fever.The auxiliary examination of the first hospital displayed, cerebrospinal fluid: sugar qualitative (+ ), white blood cell count 32×10 6/L, albumin measurement (immune turbidity method) 317.00 mg/L, immunoglobulin IgG 45.80 mg/L.Herpes simplex virus (+ ). Skull magnetic resonance imaging showed: abnormal signal at the top of the frontal frontotemporal, considering intracranial infection.Video electroencephalogram: the background is diffuse slow wave, a small amount of multifocal spikes, sharp waves, spine slow wave release, left frontal, and temporal sacral protrusions.One partial seizure may be detected during the awake period.The diagnosis was " herpes simplex virus encephalitis" , and the body temperature of the child returned to normal after anti-infection and hormone therapy.However, there were still cognitive impairments, irritability, and no language communication.After 2 years, there was no abnormality in routine biochemical and viral cerebrospinal fluid examination.Serum and cerebrospinal fluid autoimmune encephalitis-related antibody spectrum: anti-NMDA, AMPA1/2, GABAB receptor antibody and anti-CASPR2, LGI1 antibodies were negative in serum.The anti-AMPA2 receptor antibody in the cerebrospinal fluid was weakly positive, and the final diagnosis was anti-AMPA2 receptor antibody encephalitis.After the application of hormones, the children′s cognition improved, mood was more stable than before, and language communication improved as well.Anti-AMPA2 receptor antibody encephalitis can be observed in children, and may be related to immune response after viral infection.For patients of viral encephalitis with poor treatment or disease relapse and progression, the possibility of autoimmune encephalitis should be considered.

Genetic susceptibility genes of childhood moyamoya disease / 国际儿科学杂志

Moyamoya disease(MMD) is a chronic progressive cerebrovascular disease characterized by progressive stenosis or occlusion of bilateral infernal of internal carotid artery, initial segments of anterior cerebral artery and middle cerebral artery and abnormal vascular network formation accompanied by compensatory hyperplasia of skull base.With the development of neuroimaging technology, the report of the disease has a rising trend year by year, the etiology and pathogenesis of MMD have not been clarified yet.Because of its racial susceptibility and familial aggregation, it is believed that genetic factors may be involved in the pathogenesis.This article mainly reviews the genetic related genes of MMD, aiming at understanding the role of genetic related genes in the occurrence and development of MMD in detail and comprehensively so as to make diagnosis earlier and more accurately, which is helpful for timely treatment and improvement of long-term prognosis for children.

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.

The management of antiepileptic drugs in women during lactation / 中华实用儿科临床杂志

Epilepsy is one of the most common chronic diseases of the central nervous system,many epilepsy patients need lifelong medication.Epidemiological studies have shown that 3‰-5‰ neonates born by women suffering from epilepsy.Treatment become more challenging because not only the patients themselves but also the breastfed in fants should be taken into consideration.This paper reviewed how to choose lactation antiepileptic drug.In short,choosing drugs which transport less to milk and have less side effects to infants,using the lowest effective dose and avoiding combination if possible can ensure the safety of breastfeeding.

Change and significance of the soluble triggering receptor expressed of myeloid cells-1 and soluble urokinase plasminogen activator receptor expression in children with sepsis / 中华实用儿科临床杂志

Objective To investigate the change and clinical significance of soluble triggering receptor expression of myeloid cells-1 (sTREM-1) and soluble urokinase plasminogen activator receptor (suPAR) expression in children with sepsis.Methods There were 80 systemic inflammatory response syndrome (SIRS)patients who were included in the study,60 cases in the sepsis group,20 cases in the non-infectious SIRS group and 30 cases in the healthy control group.By using the enzyme-linked immunosorbent assay (ELISA)to dynamically monitor the levels of serum sTREM-1,suPAR in children with sepsis,the differences of sTREM-1,suPAR levels between children with sepsis and non-sepsis were observed,the correlation with the pediatric critical illness score(PCIS) was analyzed,and the sensitivity and specificity of sTREM-1,suPAR,C-reactive protein (CRP)and procalcitonin (PCT)and other biochemical markers were compared,and the value of sTREM-1,suPAR,CRP,PCT in the early determination and prognosis of sepsis were investigated.Results Serum sTREM-1,suPAR,PCT levels in sepsis group were significantly higher than non-infectious SIRS group and the healthy control group,and the difference was statistically significant (P ＜ 0.05),but the differences of serum CRP levels in non-infectious SIRS group and sepsis group were not statistically significant(P ＞ 0.05).In sepsis subgroup,serum sTREM-1,suPAR,PCT levels between the three groups were of statistically significant difference (P ＜ 0.05).Through dynamic monitoring of sepsis group,serum sTREM-1,suPAR,CRP,PCT levels had a gradual downward trend in 1,4,7 day,at each time point difference was statistically significant (P ＜ 0.05).Serum sTREM-1,suPAR levels in sepsis group had significant negative correlation with PCIS (r =-0.322,-0.333,P ＜ 0.05).The sensitivity and specificity of sTREM-1,suPAR,CRP,PCT on diagnosing sepsis were in a descending order,and sTREM-1 combined with suPAR has the highest sensitivity and specificity.Conclusions sTREM-1 and suPAR all can serve as indicators of infection and inflammation,as their expression level can reflect the severity of sepsis.sTREM-1 combined with suPAR diagnostic sensitivity and specificity of sepsis was significantly better than a single indicator of sTREM-1,suPAR,CRP,PCT.Combining multiple indicators can improve the accuracy of diagnosis.

Report one child with idiopathic hypereosinophilic syndrome / 临床儿科杂志

Objective To discuss the diagnosis and treatment of idiopathic hypereosinophilic syndrome (IHES) in children. Method The course and treatment process of a 6-year-old child with IHES had been retrospectively analyzed. Result The boy was admitted for abdominal discomfort and poor appetite, quickly developed into abdominal distension, dyspnea, jaundice, edema, and worsen hepatosplenomegaly. Routine blood test showed that the eosinophilia was 186.39×109/L. Bone marrow smear showed that the mature eosinophilcell granulocyles signiifcantly increased to 90.4%. The FIL1P1-PDGFRαfusion gene detection, parasites and antibodies tests were all negative. CT and other examinations indicated that the digestion, circulation, blood and nervous system were all affected. The diagnosis of IHES was considered. Hydroxycarbamide and steroids applied, the eosinophil decrease, however, the symptoms no relief, eventually developed to the multiple organ failure. Conclusion IHES is rare in children. Further studies are necessary regarding the treatment and prognosis.

Clinical analysis of two cases of epidermal nevus syndrome and a review of literature / 中华全科医师杂志

Clinical, radiological and pathological presentations in two children with epidermal nevus syndrome were analyzed and relevant literature was reviewed.Two patients both had typical epidermal nevus and abnormal cerebral radiography, which was associated with mental retardation, epilepsy, language and movement retardation.One case was complicated with an ocular tumor.Pathological investigations of the epidermal nevus revealed papilliform proliferation in squamous epidermis.The disorder may have a systemic involvement besides cutaneous lesions, with a predilection for the central nervous system.Early diagnosis and therapy may help to improve patients life quality.

Enhanced expression of aquaporin-9 in rat brain edema induced by bacterial lipopolysaccharides / 华中科技大学学报（医学）（英德文版）

To investigate the role of AQP9 in brain edema, the expression of AQP9 in an infectious rat brain edema model induced by the injection of lipopolysaccharide (LPS) was examined. Immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that the expressions of AQP9 mRNA and protein at all observed intervals were significantly increased in LPS-treated animals in comparison with the control animals. Time-course analysis showed that the first signs of blood-brain barrier disruption and the increase of brain water content in LPS-treated animals were evident 6 h after LPS injection, with maximum value appearing at 12 h, which coincided with the expression profiles of AQP9 mRNA and protein in LPS-treated animals. The further correlation analysis revealed strong positive correlations among the brain water content, the disruption of the blood-brain barrier and the enhanced expressions of AQP9 mRNA and protein in LPS-treated animals. These results suggested that the regulation of AQP9 expression may play important roles in water movement and in brain metabolic homeostasis associated with the pathophysiology of brain edema induced by LPS injection.

The effect of docosahexaenoic acid and 5-fluorouracil on human gastric cancer cell lines and the expression of bcl-2, bcl 2l12 and bax / 中华普通外科杂志

Objective To evaluate the growth inhibition of human gastric carcinoma cell lines SGC 7901 in vitro and the expression of bcl-2, bcl 2l12 and bax with docosahexaenoic acid (DHA) and 5-fluorouracil (5-FU). Methods The effect of DHA and 5-FU was measured by trypan blue, and the interaction between two agents was judged by combination index (CI). Cells were observed by inverted microscope. Flow cytometry was used for analysis of apoptosis by PI staining and Annexin-V/PI. RT-PCR was used to analyze the levels of bcl-2, bcl 2l12 and bax mRNA. Results DHA significantly inhibited the growth of SGC 7901 cells in a dose- and time-dependent way ( P < 0. 05 ), the IC50 of 24 h and 48 h was 67. 81 μg/ml and 45.76 μg/ml, and a strong synergism was found in the combination of DHA and 5-FU (CI < 1 ,P <0. 01 ). Treated by DHA and 5-FU for 48 h, cells became sparse under inverted microscope. DHA or 5-FU was able to induce apoptosis and the effect became even more significant by the combination of DHA and 5-FU. Cells were holted in phase of G01/G1 and S. RT-PCR showed that DHA or 5-FU down-regulated the expression of bcl-2 and bcl 2l12 mRNA, while bax mRNA expression was not downregnlated. Conclusions DHA could inhibit the growth of gastric carcinoma cells, DHA and 5-FU had synergetic effect in the inhibition of the cells growth and blockage of the cell cycles possibly by down-regulating the expression of bcl-2 and bcl 2l12.

Up-regulation of mRNA expressions of fas,bcl-2,bim,bax,caspase-3,caspase-9,and bcl2l12 in K562 treated with bortezomib / 中国病理生理杂志

AIM:To detect the treatment of K562 leukemia cells with bortezomib altering the expression of genes fas,bcl-2,bcl2l12,bim,bax,caspase-9 and caspase-3.METHODS:MTT assay was used to detect the inhibition of proliferation.Apoptosis was detected by Annexin-V staining and mitochondrial transmembrane potential(??m).RT-PCR was used to analyze the mRNA expressions of fas,bcl-2,bcl2l12,bim,bax,caspase-3 and caspase-9.RESULTS:Bortezomib caused a time-and dose-dependent inhibition of cell proliferation and IC50 of 24 h and 48 h were 161.41 nmol/L and 96.33 nmol/L,respectively.At the concentration of 104 nmol/L,bortezomib induced apoptosis in a time-dependent manner,including increasing annexin-V positivity and decreasing the ??m.RT-PCR showed that bortezomib up-regulated the mRNA expression of fas,bcl2l12,caspase-9 and caspase-3,but mRNA expressions of bcl-2,bim and bax did not changed obviously.CONCLUSION:Bortezomib inhibits the proliferation of K562 and induces apoptosis,in which fas,bcl2l12,caspase-9 or caspase-3 gene is one of the main genes taking part in.