Analysis of clinical phenotype and gene variation of pyridoxine-dependent epilepsy caused by ALDH7A1 gene mutation / 中华神经科杂志

Objective:To analyze the clinical phenotype and genotype characteristics of children with pyridoxine-dependent epilepsy (PDE) and provide evidence for diagnosis.Methods:Clinical data of 3 children with PDE enrolled in the Department of Neurology of Hunan Children′s Hospital from July 2016 to December 2020 were collected, and whole-exome sequencing (WES) was used for analysis. Pathogenic variants were analyzed and screened using bioinformatics tools combined with clinical phenotype. Sanger sequencing was used to analyze the source of mutations in children′s core family members.Results:Cases 1 (female) and 2 (male) were siblings, both of whom had convulsions within 24 hours after birth. WES results showed that the siblings carried compound heterozygous mutations of c.796C>T (p.R266 *) and c.1553G>C (p.R518T) in the ALDH7A1 gene, coming from the father and mother of the siblings respectively. Both of the mutations have been reported as pathogenic. Case 3, female, developed convulsions at the age of 1. WES results revealed that she carried compound heterozygous mutations of c.1094-109T>A and c.7C>T (p.R3C) in the ALDH7A1 gene, coming from her father and mother respectively. After searching HGMDPro, PubMed, 1000 Genomes, and dbSNP databases, both of the 2 mutations of c.1094-109T>A and c.7C>T (p.R3C) were not reported. The pathogenicity predictions of the 2 mutations were carried out by different biological information analysis software. The results showed that both of the mutations were harmful. All the 3 children had no epileptic seizures after treatment with increased doses of vitamin B6. Conclusions:When infants have unexplained convulsions, especially in the neonatal stage, PDE caused by ALDH7A1 gene mutation should be considered. Pyridoxine precision treatment has a good effect. The 2 de novo mutations of c.1094-109T>A and c.7C>T (p.R3C) enrich the mutation spectrum in the ALDH7A1 gene. WES has the auxiliary significance in the diagnosis of epilepsy.

Glyceryl phenylbutyrate in the treatment of argininosuccinate synthetase deficiency: a case report / 中华实用儿科临床杂志

Clinical data of a child with high blood ammonia and suspected argininosuccinate synthetase deficiency (ASSD) in Hunan Children′s Hospital were retrospectively analyzed, including data of mass spectra for blood amino acids and acyl carnitine, urine organic acid analysis and whole exome sequencing.After the exact diagnosis of ASSD and being approved by the Administrative Regulation for Import Medical Devices Urgently Needed in Boao Lecheng International Medical Tourism Pilot Zone of Hainan Free Trade Port, the patient was medicated with Glyceryl phenylbutyrate (GPB) and followed up.The patient was a boy aged 7 years and 8 months, who presented at the Neurology Department of Hunan Children′s Hospital for sleepiness, abnormal mental behavior and personality change for 1 week on December 2, 2021.Before GPB treatment, the highest blood ammonia, alanine aminotransferase and aspartate transaminase were 325.2 μmol/L, 465.7 IU/L and 277.3 IU/L, respectively.Genetic metabolism assay of blood and urine showed a significantly increased citrulline at 697.42 μmol/L; urine organic acid analysis showed increased urinary orotic acid at 144.2 μmol/L, and increased uracil at 65.1 μmol/L.A pure heterozygous variant of the ASS1 gene (c.1087C>T, p.R363W) was detected.After GPB treatment, the blood ammonia levels were 21.3 μmol/L, 54.6 μmol/L and 62.4 μmol/L on the 41 st, 90 th and 146 th days, respectively.Until July 20, 2022 follow-up visit, the patient recovered well without adverse events.This was the first ASSD child in China who was treated with GPB.This case report provided therapeutic experience of ASSD in our country.ASSD has a high mortality rate and unexplained abnormal mental behavior.It is necessary to timely measure blood ammonia, and a series of urea cycle disorders should be well concerned.The diagnosis and management of ASSD rely on the data of metabolism examination and genetic testing.

Analysis of clinical phenotype and genetic variants in a child with mitochondrial F-S disease due to variants of FDXR gene / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical phenotype and genetic variants of a child suspected for mitochondrial F-S disease.@*METHODS@#A child with mitochondrial F-S disease who visited Department of Neurology, Hunan Provincial children's Hospital on November 5, 2020 was selected as research subject of this study. Clinical data of the child was collected. The child was subjected to whole exome sequencing (WES). Bioinformatics tools were used to analyze the pathogenic variants. Candidate variants were verified by Sanger sequencing of the child and her parents.@*RESULTS@#WES revealed that the child has harbored compound heterozygous variants of the FDXR gene, namely c.310C>T (p.R104C) and c.235C>T (p.R79C), which were inherited from her father and mother, respectively. Neither variant has been reported in HGMD, PubMed, 1000 Genomes, and dbSNP databases. Both of the variants have been suggested as deleterious according to the prediction results from different bioinformatics analysis software.@*CONCLUSION@#Mitochondrial diseases should be suspected for patients with multiple system involvement. The compound heterozygous variants of the FDXR gene probably underlay the disease in this child. Above finding has enriched the spectrum of FDXR gene mutations underlying mitochondrial F-S disease. WES can facilitate the diagnosis of mitochondrial F-S disease at the molecular level.

Genetic analysis of a child with early onset neurodevelopmental disorder with involuntary movement and a literature review / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical phenotype and genetic basis of a child with early onset neurodevelopmental disorder with involuntary movement (NEDIM).@*METHODS@#A child who presented at Department of Neurology of Hunan Children's Hospital on October 8, 2020 was selected as the study subject. Clinical data of the child were collected. Genomic DNA was extracted from peripheral blood samples of the child and his parents. Whole exome sequencing (WES) was carried out for the child. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. Relevant literature was searched from the CNKI, PubMed and Google Scholar databases to summarize the clinical phenotypes and genetic variants of the patients.@*RESULTS@#This child was a 3-year-and-3-month boy with involuntary trembling of limbs and motor and language delay. WES revealed that the child has harbored a c.626G>A (p.Arg209His) variant of the GNAO1 gene. Sanger sequencing confirmed that neither of his parents has carried the same variant. The variant had been reported in HGMD and ClinVar databases, but not in the dbSNP, ExAC and 1000 Genomes databases. Prediction with SIFT, PolyPhen-2, and Mutation Taster online software suggested that the variant may be deleterious to the protein function. By UniProt database analysis, the encode amino acid is highly conserved among various species. Prediction with Modeller and PyMOL software indicated that the variant may affect the function of GαO protein. Based on the guideline of the American College of Medical Genetics and Genomics (ACMG), the variant was rated as pathogenic.@*CONCLUSION@#The GNAO1 gene c.626G>A (p.Arg209His) variant probably underlay the NEDIM in this child. Above finding has expanded the phenotypic spectrum of GNAO1 gene c.626G>A (p.Arg209His) variant and provided a reference for clinical diagnosis and genetic counseling.

Application of lacosamide in sodium channel-related epilepsy in young infants / 中华神经科杂志

Objective:To report 2 young infants of sodium channel related epilepsy with SCN2A gene mutation, and to discuss the clinical characteristics of the disease and the efficacy and safety of lacosamide combined with the literature.Methods:Corresponding information of 2 children hospitalized in the Department of Neurology of Hunan Children′s Hospital in July 2021 and October 2021 was collected, including the symptoms, comprehensive physical examination, blood, cerebrospinal fluid, imaging, electrophysiological examination, diagnosis and treatment process, response to treatment and other clinical data, as well as the sequencing results of the whole exome of the children. The efficacy and safety of lacosamide were analyzed, and the related literatures of the Biomedical Literature Database, Wanfang Data Knowledge Service Platform and Chinese Knowledge Infrastructure Database were searched and reviewed.Results:Both of the 2 cases were girl. Their onset age was within 3 months. The initial symptoms were frequent convulsions and backward development. There was no structural abnormality in the head image. The convulsions could not be controlled according to conventional multidrug treatment. The seizures were quickly controlled with lacosamide. Now they have been followed up for 6 months. No obvious adverse reactions were found. Case 1 gene test results showed the SCN2A gene (chr2:166152333-166246334) heterozygous deletion, SCN1A gene (chr2:166847754-16693013) heterozygous deletion, the deletion size being about 5.72 Mb. Case 2 gene test results showed new missense mutation of SCN2A (c.1285G>A, p.Glu429Lys). There were dozens of seizures every day. They were treated with valproic acid, oxcarbazepine and levetiracetam successively. The seizures could not be controlled. Three focal seizures originated in the left temporal region were detected by electroencephalogram. There was no recurrence on the third day after adding lacosamide, and there was no attack after 5 months of follow-up. No obvious adverse reactions were found during follow-up.Conclusions:Sodium channel related epileptic encephalopathy often starts early, has frequent seizures, and can be accompanied by backward psychomotor development at the same time. The slow sodium channel blocker lacosamide has good efficacy and safety in the treatment of sodium channel-related epilepsy with SCN2A gene mutation or combined SCN1A gene mutation.

Factors predicting the risk of seizure relapse after antiepilepsy drug withdrawal in childhood epilepsy / 中华实用儿科临床杂志

Objective To investigate the factors associated with seizure relapse after antiepilepsy drug (AED) withdrawal in childhood epilepsy.Methods A retrospective analysis was conducted in epileptic children of Hunan Children's Hospital from Jan.2003 to Jan.2011.Among those with anti-epileptic therapy for seizure-free period over 2 years,the patients who relapsed after withdrawal were followed up through outpatient clinic visits and/or by telephone interviews for at least 2 years.Results Of the 127 cases of children enrolled in this study,28 patients(22.05％) relapsed [male:12/59 cases (20.34％) and female:16/68 cases (23.53％)].Cumulative relapse rates were 18.18％ (8/44 cases) in infancy,15.79％ (6/38 cases) in toddlers,23.53％ (8/34 cases) in preschool children,and 54.55％ (6/11 cases)in school age group.Of the patients who relapsed,generalized seizure occurred in 12/87 cases (13.79％),partial seizure in 16/40 cases(40.00％).According to seizure frequency between the first seizure and AED administration,3 cases(6.25％) relapsed among 48 cases of seizure frequency ＜ 5 times,13 cases(24.07％) relapsed among 54 cases of seizure frequency 5 to 10 times,and 12 cases(48.00％) relapsed among 25 cases of seizure frequency more than 10 times.Relapse occurred in 9 cases of monotherapy(9/91 cases,9.89％) and in 19 cases of polytherapy (19/36 cases,52.78％).According to the seizure control period (period between the beginning of antiepileptic treatment and AED withdrawal),14 cases relapsed among 37 cases with the seizure control period of 2 to 3 years (37.84％),8 cases relapsed among 51 cases with the period of 3 to 4 years (15.69％),and 6 cases relapsed among 39 cases with the period of 4 to 5 years(15.38％).According to AED tapering off period,10 cases relapsed among 24 cases with the period of 3 months (41.67％),9 cases relapsed among 36 cases with the period of 3-6 mc ths (25.00％),and 9 cases relapsed among 67 cases with the period of over 6 months(13.43％).Factors associated with an increased risk of relapse were age of epilepsy onset,seizure type,route of administration,timing of antiepileptic trug withdrawal,tapering speed,which were had statistical significance (x =8.051,6.780,16.896,27.607,7.576,8.451,all P ＜0.05).Gender difference was not associated with the risk of relapse(x2 =0.187,P ＞ 0.05).Conclusions Factors associated with an increased risk of relapse are age of epilepsy onset,seizure type,route of administration,timing of antiepileptic drug withdrawal,tapering speed.Standard therapies of early treatment,adherence to medication for at least 3 years,taper period for more than 6 months are associated with a decreased probability for relapse.

Changes of Caveolin-1, IL-1β, VEGF in cerebrospinal fluid of children with viral encephalitis and their clinical significance / 国际儿科学杂志

Objective To detect the disparity of three biological molecules Caveolin-1,IL-1β,VEGF in cerebrospinal fluid of children with viral encephalitis at the different stages; to explore the role of Caveolin-1,IL-1β,VEGF in the pathogenesis of viral encephalitis;and to evaluate their clinical significance in assessing the severity and prognosis of viral encephalitis.Methods We recruited 65 inpatients children with viral encephalitis in the Second Neurology Department of Hunan Children's Hospital from July 2011 to July 2012.Subjects were divided into 2 groups:54 cases of acute phase and 11 cases of recovery phase.According to the clinical manifestations,they were re-divided into 40 patients with mild viral encephalitis and 25 cases of severe viral encephalitis.Twenty healthy age matched controls (10 cases of epilepsy and 10 cases of congenital abnormality) were also taken for the study.Cerebrospinal fluid exam,EEG,head MRI and other tests were performed in all patients.Caveolin-1,IL-1β and VEGF levels in cerebrospinal fluid of 65 children with viral encephalitis and 20 age-matched controls were measured using ELISA.Results Cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels in the acute phase of viral encephalitis were (49.209 ± 22.320) pg/ml,(16.923 ± 6.823) ng/ml,(44.342 ± 19.264) ng/ml respectively,and (33.253 ± 20.349)pg/ml,(11.724 ± 3.009)ng/ml,(30.312 ± 18.147) ng/ml in recovery phase,which were significantly higher than those of controls (P ＜0.01).The difference was statistically significant between acute phase and recovery phase (P ＜ 0.05).Acute viral encephalitis patients had higher Caveolin-l,IL-1β,VEGF levels than the epilepsy group,and the difference was statistically significant (P ＜ 0.05).In viral encephalitis group,children with cerebrospinal fluid protein content (0.5 ～ 1.0 g / L) had higher of Caveolin-1,IL-1β and VEGF levels as compared with those with cerebrospinal fluid protein content ≤ 0.5 g/L,and the difference was statistically significant (P ＜ 0.01).Cerebrospinal fluid Caveolin-1,IL-1 β and VEGF showed no significant difference among children with different severity of encephalitis,different levels of frequent seizures,different degrees EEG changes (P ＞ 0.05).But in the patients with severe head MRI changes,cerebrospinal fluid Caveolin-1,IL-1β,VEGF levels increased significantly (P ＜ 0.05).Conclusions Caveolin-1,IL-1β and VEGF may participate in the pathogenesis of viral encephalitis.Detection of these parameters may be helpful to the evaluation of the severity and prognosis of viral encephalitis.

Effect of health education on farmers' intention of joining in the new rural cooperative medical system / 北京大学学报(医学版)

Objective:To evaluate the impact of health education intervention on promoting rural residents to join new rural cooperative medical system(NCMS)and their intention of joining NCMS based on Health Belief Model in project areas in Henan and Jilin Provinces.Methods:Quasi-experiment study was used to evaluate intervention impact.Following the evidence-based approach,according to needs assessment,a half-year health education intervention was implemented among farmers in the experimental counties in Henan and Jilin Provinces respectively.A questionnaire survey was conducted among farmers in intervention and control counties before and after intervention,and intervention impact was evaluated by comparing the indicators' changes in intervention and control counties.Results:After health education intervention,the knowledge level of farmers in two intervention counties increased by 29.0% and 37.8% respectively,their scores of perceived threatens of health risk and perceived barriers of joining NCMS among the respondents were decreased.Meanwhile,their score of perceived benefit of joining NCMS were increased,and the rate of willingness to join NCMS increased remarkably in both intervention counties.Conclusion:Health education was effective and helpful in increasing farmer's knowledge,understanding and cognitive level of NCMS,and it should play an important role for the sustainable development of NCMS.

STUDY ON EXTRACTION AND BIOLOGICAL ACTIVITY OF ANTIOXIDANT SUBSTANCE OF SWEET POTATOES / 营养学报

Objective: The chemical composition, extraction and biological activity of antioxidant substance of sweet potatoes were studied. Method: The antioxidant substance was extracted with alcohol and purified with AB-8 big-hole resin and its elimination rate of _?O 2 and OH and SOD activity were determined.Results: The content of antioxidant substance stockpiled respectively in the root tubers, stems and leaves of sweet potatoes and was different in varieties. The highest content in the root tuber was 0.324%, which accounted 70% of total content in sweet potatoes. Its extraction ratio was 85%. The major composition of the antioxidant substance was phenol acid, including chlorogenic acid,isochlorogenic acid,neochlorogenic acid and 4-0-caffeonyl quinic butyl ester. They had intense antioxidant function and their elimination rate to _?O 2 and?OH at concentration 90 mg/ml was 88.02% and 82.21% respectively. The antioxidantsubstance restrained oxidation of lipid and hemolysis of red cell induced by H2O2,and increased SOD activity in serum and skin of mice. Conclusion: Several kinds of phenol acid as antioxidant substance stockpiled in sweet potatoes were proved to have strong antioxidant activity.

FRACTIONATION OF POLYSACCHARIDBS FROM CURCUMA ZEDOARIA ROSC AND ITS BIOACTIVITIES / 营养学报

Objective: To extract the polysaccharides of Curcuma zedoaria Rosc (PCZ) and study its bioactivities. Methods: The crude polysaccharides were extracted from herbal slice of Curcuma zedoaria Rosc with hot water ,and precipitated by ethanol and savage deproteinization and depigmentation with activated charcoal. The constituents were isolated by column chromatography and the total polysaccharides content was determined. Experiments of immune function,anti-oxidation and anti-tumor activity of KM mice and in viro were studied. Results: The content of total polysaccharides of Curcuma zedoaria Rosc was 33.12%. and the extraction rate was 1.938% . It could raise spleen cell proliferation and transformation rate of lymphocyte and SOD activity in blood of mice. A higher antitumor effect of the polysaccharides by injection was observed dose-dependently. Conclusion: PCZ could raise the immune and anti-oxidative activities and then had a higher antitumor effect.

STUDY ON THE ANTIOXIDANT ACTIVITY OF SWEET POTATO ANTHOCYANIN AND ITS INHIBITING EFFECT ON GROWTH OF CANCER S_(180) / 营养学报

Objective: To study the antixiodant activity of sweet-potato anthocyanin (SPAC) and its inhibiting effect on growth of cancer S180. Method: The scavenging capacity of ??O2 and?OH and the inhibitory effect of MDA were determined in vitro.The SOD activity of serum and skin, the effect on S180 growth, serum GSH-Px activity and MDA were tested in mice. Results: SPAC had antioxidant activity in vitro, with elimination rate of O2?? and?OH 30.63% and 20.57% higher than the same concerntration of VC respectively. Activity of SOD in the serum and skin of mice were raised 27.30% and 13.50% respectively after giving SPAC. The growth rate of cancer S180 in the mice was inhibited, and the highest inhibitory rate was 43.12%. The activity of serum GSH-Px, SOD of the mice was raised as compared to control, but MDA declined . Conclusion: SPAC is a perfect natural pigment with inhibitory effect on growth of cancer S180 probably through its antioxidant activity.

PROPERTIES OF AN ANTIOXIDANT FROM ALOE WITH PROTECTIVE EFFECT ON RED BLOOD CELLS / 营养学报

Objective: To investigate the free radical scavenging properties of six aloe species and study its protective effect on red blood cells. Methods: Free radical scavenging properties were investigated in modified chemical system. The antioxidant was purified by silica column and thin layer chromatography and analyzed by HPLC. Its protection on red blood cells was studied with red blood cells culture system in vitro. Results: The extract from A. berebihoria showed the most effective scavenging activities on O - 2? and ?OH (32.19% and 26.05% respectively) in six aloe species. The fraction A with a Rf value of 0 497 was proved to be a pure antioxidant (Rt=15.956 min). It is an anthraquinone with phenol hydroxyls. Fraction A showed strong inhibitory activity on H 2O 2 induced hemolysis of red blood cells and its IC 50 was 0.856 g/L. Conclusion: Fraction A of the extract from A. berebihoria had strong scavenging activities of O - 2? and ?OH, thereby inhibiting the peroxidation of red blood cells and maintaining their functions.