Construction and application of rational drug use database for children in our hospital / 中国药房

OBJECTIVE To establish a database of rational drug use for children in our hospital， and to provide reference for ensuring the safety of drug use in children. METHODS The construction and filling of the knowledge base of rational drug use for children were performed by establishing the basic structure of the knowledge base， formulating reference standards for the quality level of pediatric medication evidence， and refining evidence-based evidence of pediatric medication. The rule base of rational drug use for children was designed and built from four aspects： preliminary determination rules for patient information， basic drug information rules， prescription suitability review rules， result labeling and post-processing rules. The database was embedded into prescription review system of our hospital and was applied online to test its effectiveness. RESULTS A set of database containing 672 commonly used pediatric medicines and more than 15 000 rules for rational drug use for children was initially constructed. The average interception rate of unreasonable medical orders for hospitalized children after database application（from December 2021 to May 2022）was 2.03%， and was higher than 0.80% before database application（from June 2021 to November 2021）（χ 2＝5 784.389， P＜0.001）； after post-sampling and prescription review， the average qualified rate of medication orders in discharged medical records for children after the application of the database was 99.10%， and was higher than 94.58% before the application of the database （χ 2＝301.237， P＜0.001）. CONCLUSIONS Self-constructed evidence-based rational drug use database for children is close to the actual clinical needs of pediatrics in medical institutions， which can effectively reduce clinical irrational drug use behaviors in pediatrics， improve the pass rate of prescriptions， and ensure the safety of children’s drug use.

A case of dilated cardiomyopathy caused by FHL2 gene variant and a literature review / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical phenotype and genetic features of a child with dilated cardiomyopathy (DCM).@*METHODS@#Clinical data of the child who had presented at the Zhengzhou Children's Hospital on April 28, 2020 was collected. Trio-whole exome sequencing (trio-WES) was carried out for the child and her parents, and candidate variants were validated by Sanger sequencing. "FHL2" was taken as the key word to retrieve related literature from January 1, 1997 to October 31, 2021 in the PubMed database and was also searched in the ClinVar database as a supplement to analyze the correlation between genetic variants and clinical features.@*RESULTS@#The patient was a 5-month-old female infant presented with left ventricular enlargement and reduced systolic function. A heterozygous missense variant c.391C>T (p.Arg131Cys) in FHL2 gene was identified through trio-WES. The same variant was not detected in either of her parents. A total of 10 patients with FHL2 gene variants have been reported in the literature, 6 of them had presented with DCM, 2 with hypertrophic cardiomyopathy (HCM), and 2 with sudden unexplained death (SUD). Phenotypic analysis revealed that patients with variants in the LIM 3 domain presented hypertrophic cardiomyopathy and those with variants of the LIM 0~2 and LIM 4 domains had mainly presented DCM. The c.391C>T (p.Arg131Cys) has been identified in a child with DCM, though it has not been validated among the patient's family members. Based on the guidelines of the American College of Medical Genetics and Genomics, the c.391C>T(p.Arg131Cys) variant was re-classified as likely pathogenic (PS2+PM2_Supporting+PP3+PP5).@*CONCLUSION@#The heterozygous missense variant of c.391C>T (p.Arg131Cys) in the FHL2 gene probably predisposed to the DCM in this child, which has highlighted the importance of WES in the clinical diagnosis and genetic counseling.

Clinical and genetic analysis of eight children with Primary hypertrophic cardiomyopathy / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical and genetic characteristics of eight children with Primary hypertrophic cardiomyopathy (HCM).@*METHODS@#Eight children with HCM admitted to the Department of Cardiology of Henan Children's Hospital from January 2018 to December 2021 were selected as the study subjects. Clinical data of the children were collected. Whole exome sequencing was carried out on two children, and trio whole exome sequencing was carried out on the remainder 6 children. Sanger sequencing was used to verify the candidate variants in the children and their parents, and the pathogenicity of the variants was evaluated based on the guidelines from the American College of Medical Genetics and Genomics (ACMG).@*RESULTS@#The patients had included 5 males and 3 females, with their ages ranging from 5 to 13 years old. The average age of diagnosis was (7.87 ± 4.8) years old, and the cardiac phenotype showed non-obstructive HCM in all of the patients. WES has identified variants of the MYH7 gene in 4 children, including c.2155C>T (p.Arg719Trp), c.1208G>A (p.Arg403Gln), c.1358G>A (p.Arg453His), and c.1498G>A (p.Glu500Lys). Based on the guidelines from the ACMG, the first 3 variants were classified as pathogenic, while c.1498G>A (p.Glu500Lys) was classified as likely pathogenic (PM1+PM2_Supporting+PM6+PP3), which was also unreported previously. The remaining four children had all harbored maternal variants, including MYL2: c.173G>A (p.Arg58Gln; classified as pathogenic), TPM1: c.574G>A (p.Glu192Lys) and ACTC1: c.301G>A (p.Glu101Lys)(both were classified as likely pathogenic), and MYBPC3: c.146T>G (p.Ile49Ser; classified as variant of uncertain significance). Seven children were treated with 0.5 ~ 3 mg/(kg·d) propranolol, and their symptoms had improved significantly. They were followed up until September 30, 2022 without further cardiac event.@*CONCLUSION@#Genetic testing can clarify the molecular basis for unexplained cardiomyopathy and provide a basis for clinical diagnosis and genetic counseling. Discovery of the c.1498G>A (p.Glu500Lys) variant has also expanded the spectrum of MYH7 gene mutations underlying HCM.

Genetic analysis of a child with restricted cardiomyopathy and phenylketonuria and a literature review / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical and genetic characteristics of a child with restricted cardiomyopathy (RCM) and phenylketonuria (PKU), and summarize the clinical characteristics and genetic diversity of RCM in children through a literature review.@*METHODS@#A child with RCM in conjunct with PKU who was admitted to the Children's Hospital Affiliated to Zhengzhou University in June 2020 due to edema of eyelids and lower limbs for 1 year and aggravation for over 1 month was selected as the study subject. Relevant clinical data were collected. Peripheral blood samples of the child and his parents were collected for whole exome sequencing (WES). Candidate variants were validated by Sanger sequencing and bioinformatic analysis. Childhood, TNNI3 gene and restricted cardiomyopathy were used as the keywords to search the Wanfang data knowledge service platform, Chinese Journal Full-text database and PubMed database, and the search period was limited to from the time of establishment till August 2022. Clinical manifestations and characteristics of the TNNI3 gene variants were summarized.@*RESULTS@#The child, a 2-year-old-and-4-month-old male, had normal intelligence, facial features and normal hair and skin color, but his motor and physical development was delayed, in addition with edema of bilateral eyelids and lower limbs. The results of WES and Sanger sequencing revealed that he has harbored compound heterozygous variants of the PAH gene, namely c.331C>T (p.R111X) and c.940C>A (p.P341T), which were inherited from his father and mother, respectively. In addition, he has also harbored a de novo heterozygous variant of c.508C>T (p.R170W) of the TNNI3 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the TNNI3: c.508C>T (p.R170W) was classified as a pathogenic variant (PS2+PS4+PM2_Supporting+PM5), PAH: c.331C>T (p.R111X) as a pathogenic variant (PVS1+PM2_Supporting+PM3+PP4), and c.940C>A (p.P341T) as a likely pathogenic variant (PM2_Supporting+PM3+PM5+PP4). In total 30 children with RCM caused by TNNI3 gene variants were retrieved, with a male-to-female ratio of 1 : 1.55 and manifestations including heart failure, sinus rhythm, bi-atrial enlargement, ST-T wave change, ventricular restricted filling, and decreased ventricular diastolic function. In total 16 variants of the TNNI3 gene were identified, among which c.575G>A was the most common, and all cases had conformed to an autosomal dominant inheritance.@*CONCLUSION@#Phenylalanine hydroxylase deficiency and RCM are rare diseases with complex clinical manifestations. The PAH: c.331C>T (p.R111X)/c.940C>A (p.P341T) and TNNI3: c.508C>T (p.R170W) variants probably underlay the RCM and PKU in this child.

Clinical and genetic analysis of five children with Catecholaminergic polymorphic ventricular tachycardia due to variants of RYR2 gene / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical and genetic characteristics of five children with Catecholaminergic polymorphic ventricular tachycardia (CPVT).@*METHODS@#Five children with clinical manifestations consistent with CPVT admitted to the Department of Cardiology of Children's Hospital Affiliated to Zhengzhou University from November 2019 to November 2021 were selected as the study subjects. Their clinical data were collected. Potential variants were detected by whole exome sequencing, and Sanger sequencing was used to verify the candidate variants. All patients were treated with β-blocker propranolol and followed up.@*RESULTS@#All patients had developed the disease during exercise and presented with syncope as the initial clinical manifestation. Electrocardiogram showed sinus bradycardia. The first onset age of the 5 patients were (10.4 ± 2.19) years, and the time of delayed diagnosis was (1.6 ± 2.19) years. All of the children were found to harbor de novo heterozygous missense variants of the RYR2 gene, including c.6916G>A (p.V2306I), c.527G>C (p.R176P), c.12271G>A (p.A4091T), c.506G>T (p.R169L) and c.6817G>A (p.G2273R). Among these, c.527G>C (p.R176P) and c.6817G>A (p.G2273R) were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.527G>C (p.R176P) was classified as a pathogenic variant (PS2+PM1+PM2_Supporting+PM5+PP3+PP4), and the c.6817G>A (p.G2273R) was classified as a likely pathogenic variant (PS2+PM2_Supporting+PP3+PP4). The symptoms of all children were significantly improved with the propranolol treatment, and none has developed syncope during the follow up.@*CONCLUSION@#Discovery of the c.527G>C (p.R176P) and c.6817G>A (p.G2273R) variants has expanded the mutational spectrum of the RYR2 gene. Genetic testing of CPVT patients can clarify the cause of the disease and provide a reference for their genetic counseling.

Correlation between ADRB2 gene polymorphism and childhood bronchial asthma susceptibility and drug resis-tance / 中华实用儿科临床杂志

Objective To discuss the association between ADRB2 gene polymorphism of rs1042713, rs1042714 locus and susceptibility of childhood bronchial asthma and response to the treatment,and to understant the preliminary pathogenesis of asthma preliminary. Methods The oral epithelial cells of children in asthma case group from the outpatient or inpatients at the Zhengzhou Children′s Hospital (173 cases)and healthy control group (166 ca-ses)were collec-ted. The genotypes of ADRB2 gene encoding area rs1042713 locus and rs1042714 were tested by the mean of Taqman probe real-time fluorescent quantitative PCR,and the difference was analyzed. The therapeutic effects between 2 groups were compared,and then the differences in the amino acid sequence and protein structure correspon-ding genotypes of ADRB2 genes were investigated by using bioinformatics analysis. Results There were statistical differences in genotype frequency and allele frequency of rs1042713 locus between asthma case group and healthy con-trol group(P = 0. 001,0. 000),but no difference at rs1042714 locus(P = 0. 159,0. 061). GG genotype of rs1042713 locus had significant differences in adjusting the drug dosage with AA,AG genotype patients(H = 12. 583,P = 0. 002), but the different genotype of rs1042714 locus had no significant differences in adjusting the drug dosage(H = 2. 696, P = 0. 260). The polymorphism of rs1042713 locus caused protein local structure changes,but the homologous protein structure of rs1042714 locus caused no changes. Conclusions ADRB2 gene polymorphism of rs1042713 locus is not only associated with the susceptibility of childhood bronchial asthma,but also has an effect on β2-receptor agonist treatment to a certain extent. But polymorphism of rs1042714 locus may not be associated with the susceptibility of asth-ma and the response to treatment. The reason may be different genotypes leading to different protein structures and bio-logical functions. The mechanism may be related to the structural and functional differences of protein structures corre-sponding to different genotypes.

GC-MS determination of metabolites in rat kidneys / 中南大学学报(医学版)

Objective:To establish a method to determine the metabolites in rat kidney tissues by gas chromatography-mass spectrometry (GC-MS) combined with chemometric techniques. Methods:Metabolites were separated and identiifed on HP-5MS column (30 m × 0.25 μm × 0.25 mm). The initial column temperature was 100℃lasting 3 min, and then programmed at 8℃/min to 300℃, maintaining at this temperature for 6 min. hTe internal standard was heptadecanoic acid. hTe grinded kidney tissue was exacted by methanol. hTe supernatant was dried by nitrogen. Atfer the oximation and derivation, the supernatant was analyzed by GC-MS. hTe overlapped peaks were resolved into pure chromatogram and mass spectra with chemometric techniques. Qualitative analysis was performed by comparing the obtained pure mass spectra with those in NIST mass spectra database and certiifcated by the standards and the references. hTe internal method was used for semi-quantitation. Results:A total of 53 compounds were identiifed. hTe main constitutions in the kidney tissue were amino acids, saccharides, fatty acids and urea. Conclusion:hTe combination of methods is rapid and accurate for the analysis of metabolites in the kidney tissue, which provides more information for further study of metabonomics in kidney tissues.

Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro / 重庆医科大学学报

Objective:To investigate expression and role of nesprins gene during mouse embryonic stem cell(ESC)differentiation into cardiomyocyte in vitro.Methods:Mouse ESC cultrued in combining 5-azacytidine as inducer with hanging drop cultures and the differentiated cells were identified with cardiac special genes and proteins;The mRNA expression of nesprin-1 and nesprin-2 genes were detected by reverse transcription-polymerase chain reaction(RT-PCR),and sub-cellular location of nesprin-1 protein in nucleus was detected by immunofluorescence in d7,d7+3,d7+6,d7+9 and d7+20.Results:The expression of nesprin-1 and nesprin-2 genes mRNA can be found in the period of mouse ESC differentiation into cardiomyocyte.There is expression peak of nesprin-1 gene mRNA in d7,d7+3,d7+6,then declined gradually to the lowest in d7+20;expression peak of nesprin-2 gene mRNA in d7,then also declined gradually to the lowest in d7+20;The protein of nesprin-1 can be detected in the period of mouse ESC differentiation into cardiomyocyte,and mainly located in nuclear envelope and space around nuclei.Conclusion:The expression peak of nesprin-1 and nesprin-2 gene mRNA is interrelated with the phase of inducing and specialization of ESC differentiation into cardiac muscle cell,it can be speculated that nesprin genes has an important role in maintaining ESC differentiation into cardiac muscle cell.