Rapid screening of a hotspot variant c.609G>A in MMACHC gene by using PCR-high-resolution melting curve analysis / 中华医学遗传学杂志

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.

Clinical analysis of 6 cases of orbital cellulitis in neonates and literature review / 中华实用儿科临床杂志

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.

Analysis of co-segregation of methylation pattern and gene ontology among pedigrees affected with neural tube defects / 中华医学遗传学杂志

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.

Genetic analysis of one family with congenital limb malformations / 中华医学遗传学杂志

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.

Analysis of CYP21A2 gene mutation and phenotype in patients with 21-hydroxylase deficiency / 中华内分泌代谢杂志

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.

Deletional variant of REEP1 gene in a pedigree affected with spastic paraplegia type 31 / 中华医学遗传学杂志

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.

Genetic analysis of one family with congenital limb malformations / 中华医学遗传学杂志

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.

Origin and morphological features of small supernumerary marker chromosomes in Turner syndrome / 中华医学遗传学杂志

OBJECTIVE To explore the origin and morphological features of small supernumerary marker chromosomes (sSMCs) in Turner syndrome. METHODS For 5 cases of Turner syndrome with a sSMC identified by conventional G-banding, dual-color fluorescence in situ hybridization (FISH) was applied to explore their origin and morphological features. RESULTS Among the 5 cases, 3 have derived from the X chromosome, which included 2 ring chromosomes and 1 centric minute. For the 2 sSMCs derived from the Y chromosome, 1 was ring or isodicentric chromosome, while the other was an isodicentric chromosome. CONCLUSION The sSMCs found in Turner syndrome have almost all derived from sex chromosomes. The majority of sSMCs derived from the X chromosome will form ring chromosomes, while a minority will form centric minute. While most sSMC derived from Y chromosome may exist as isodicentric chromosomes, and a small number may exist as rings. For Turner syndrome patients with sSMCs, dual-color FISH may be used to delineate their origins to facilitate genetic counseling and selection of clinical regime.

Identification of a novel mutation of UPB1 gene in a Chinese family affected with beta-ureidopropinoase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular etiology for a Chinese family affected with beta-ureidopropinoase deficiency.@*METHODS@#Genomic DNA was extracted from the peripheral blood samples of family members. All exons and flanking intron regions of the UPB1 gene were amplified by PCR and detected by direct sequencing. The pathogenicity of identified mutation was analyzed using Polyphen2 and SIFT software.@*RESULTS@#Compound heterozygous mutations of the UPB1 gene, including c.853G>A (p.A285T) and c.917-1G>A, were discovered in the proband, which were inherited respectively from his mother and father. Bioinformatics analysis suggested that this novel mutation was damaging.@*CONCLUSION@#The compound heterozygous mutations of the UPB1 gene probably underlie the beta-ureidopropinoase deficiency in the infant. Discovery of c.853G>A also enriched the mutation spectrum of the UPB1 gene.

Mutation analysis on DACT1 gene in children with neural tube defects in northern Chinese Han population / 天津医药

Objective To investigate the correlation between neural tube defects (NTDs) and DACT1 gene, and provide the basic data for disease diagnosis and genetic counseling. Methods Blood samples were obtained from 163 NTDs patients and 480 unrelated healthy individuals. Mutation detection of DACT1 gene and DNA direct sequencing was carried out by PCR amplification. Bioinformatics analysis of these mutated loci was performed. Results Six mutations were found in NTDs patients, including 4 missense mutations (p.R45W, p.D142G, p.N356K and p.V702G). But these mutations were not found in 480 healthy individuals. Three mutated amino acid residues (p.45R, p.142D and p.356N) were highly conservative in evolution, and the mutated carriers were female patients, and suffered from anencephaly. Conclusion DACT1 gene mutation may be a risk factor of NTDs in Han population of northern China.

Study on DNA methylation in three neural tube defects pedigrees / 中华实用儿科临床杂志

Objective To investigate the methylation alteration of genomic DNA (gDNA) and its significance in pedigree neural tube defects (NTDs).Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group,and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted,then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification,which was put in I11umina Infinium human methylation 450k bead chip to perform hybridization,elution,extension,and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2％ of the detected CpG sites and there were 617 differential hypermethylation sites (P ＜ 0.05),and 63 of them represented significant difference(P ＜ 1 × 10-4),including zinc finger E-box binding homebox 2,5,10-methylenetetrahydrofolate dehydrogenase 1 etc;there were 104 differential hypomethylation sites (P ＜ 0.05),and 65 of them represented significant difference (P ＜ 0.01),including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients,but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree,the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.

Study on DNA methylation in three neural tube defects pedigrees / 中华实用儿科临床杂志

Objective To investigate the methylation alteration of genomic DNA (gDNA) and its significance in pedigree neural tube defects (NTDs).Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group,and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted,then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification,which was put in I11umina Infinium human methylation 450k bead chip to perform hybridization,elution,extension,and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2％ of the detected CpG sites and there were 617 differential hypermethylation sites (P ＜ 0.05),and 63 of them represented significant difference(P ＜ 1 × 10-4),including zinc finger E-box binding homebox 2,5,10-methylenetetrahydrofolate dehydrogenase 1 etc;there were 104 differential hypomethylation sites (P ＜ 0.05),and 65 of them represented significant difference (P ＜ 0.01),including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients,but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree,the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.

Identification of ALDH5A1 gene mutations in a Chinese family affected with succinic semialdehyde dehydrogenase deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutation in a Chinese family affected with succinic semialdehyde dehydrogenase deficiency.</p><p><b>METHODS</b>Genomic DNA was extracted from the peripheral blood samples of the proband, her family members and 100 healthy controls. All exons and flanking intronic regions of the ALDH5A1 gene were amplified by PCR and subjected to direct sequencing.</p><p><b>RESULTS</b>The proband was found to have compound heterozygous mutations of the ALDH5A1 gene, namely c.398_399delAA (p.N134X) and c.638G>T (p.R213L), for which her parents were both heterozygous carriers. The same mutations were not found among the 100 healthy controls.</p><p><b>CONCLUSION</b>The novel mutations of the ALDH5A1 gene probably underlie the pathogenesis of the disease in the infant, which also enriched the mutation spectrum of the ALDH5A1 gene.</p>

The clinical experience of diagnosis and treatment of late vitamin K deficiency intracranial hemorrhage as the first symptom of biliary atresia / 天津医药

Objective To investigate the surgical diagnosis and treatment of late vitamin K deficiency intracranial hemorrhage caused by biliary atresia. Methods Clinical data of six cases of biliary atresia with late vitamin K deficiency intracranial hemorrhage were collected in the Department of Neurosurgery of Tianjin Children’s Hospital from January 2000 to December 2013. Data were analyzed to identify the biliary atresia as soon as possible in the treatment of intracranial hemorrhage and prolonged jaundice in children. Results Six cases (1 male, 5 female), mean age was (16.0±2.6) days, and were treated with external drainage of intracranial hematoma and infusion therapy. In the treatment, children were found jaundice exacerbation and doubted about biliary atresia. After consultation by general surgeons, children were transferred to the department of general surgery for further treatment at an average age of (29.1±1.2) days, and were diagnosed as biliary atresia by intraoperative cholangiography. Conclusion Pediatric neurosurgeon should have a sufficient understanding and make an early diagnosis to late vitamin K deficiency intracranial hemorrhage caused by biliary atresia, to avoid delaying the optimal treatment time of biliary atresia.

Association between maternal MTHFR C677T polymorphism and neural tube defects in offsprings:a Meta-analysis / 天津医药

Objective To explore the association between maternal methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism and neural tube defects (NTDs). Methods CBM, VIP, CNKI, Wanfang, PubMed and Web of Science databases from set up to March, 2014 were electronically searched to identify case-control studies on the relationship between maternal MTHFR C677T polymorphism and NTDs. The data were quantitatively analyzed by RevMan 5.0 software. Results A total of 25 studies were selected including 2 282 cases and 3 420 controls. Overall, the pooled OR (with 95%CI) under co-dominant model and allele contrast were 2.28(1.60-3.24), 1.25(1.02-1.53) and 1.42(1.21-1.67). Subgroup analysis showed significant association between maternal MTHFR C677T polymorphism and NTDs susceptibility in Asian populations. Conclusion The present meta-analysis suggests that MTHFR C677T polymorphism is significantly associated with maternal risk for NTDs, especially in Asian populations.

The relationship between frizzled 6 gene polymorphisms and neural tube defects in children of northern Han Chinese population / 中华神经科杂志

Objective To study the association of single nucleotide polymorphisms (SNPs) of the frizzled 6(FZD6) gene with neural tube defects(NTDs) in a northern Han Chinese population.Methods Three nonsynonymous SNPs in the FZD6 gene (rs827528,rs3808553,rs12549394) were examined.The SNPs were genotyped by polymerase chain reaction (PCR) and sequencing in 135 NTD patients and matched normal controls.The allele,genotype and haplotype frequencies were calculated and analyzed to examine the association between FZD6 SNPs and NTDs.Results Both T allele and TT genotype frequencies of the rs3808553 polymorphism in the NTDs group were significantly higher than those in the controls,and children with T allele and TT genotype were associated with increased risk of NTDs (OR =1.575,95％ CI 1.112-2.230,P =0.010 and OR =2.811,95％ CI 1.325-5.967,P =0.023 respectively).There were no significant differences among different genotypes or alleles in both rs827528 and rs12549394.Haplotypes AG-C and A-T-C were found associated with NTDs in the case-control study (OR =0.560,95％ CI 0.378-0.830,P=0.004 and OR=1.670,95％CI 1.126-2.475,P =0.011 respectively).Conclusions The rs3808553 polymorphism of FZD6 is obviously associated with NTDs in children of northern Han Chinese population.The TT genotype may increase the risk for NTDs.The rs827528 and rs12549394 polymorphisms of FZD6 may have no association with NTDs.

Aneurysmal bone cyst of the calcaneus in a child: case report and review of literature / 中国矫形外科杂志

[Objective] Aneurysmal bone cyst of the ealeaneus is an uncommon entity. The purpose of the report is to describe aneurysmal bone cyst of the ealcaneus seen in a child.[ Methods] The clinical, radiological and histopathologic findings of a boy with aneurysmal bone cyst of the calcaneus were retrospectively analyzed with reference to literature. [ Results ]The tumor mainly involved the right calcaneus. He was treated successfully after the curettage and bone grafting. Histopathological examination revealed an aneurysmal bone cyst of the right calcaneus. He is now disease-free for 2 years, and no sign of recurrence was found. [ Conclusion] Aneurysmal bone cyst of the calcaneus is an uncommon entiy. To simple aneurysmal bone cyst,curettage and bone grafting are effective therapic method.

Diagnosis and treatment of split cord malformations in children / 中华全科医师杂志

We retrospectively analyzed clinical and imaging data of 26 children with split cord malformations (SCMs). Based on Pang's classification, 14 SCMs were defined as type Ⅰ and 12 as type Ⅱ.Neural function was markedly improved in 20 patients postoperatively. Three of 4 children who did not undergo surgical treatment had neural function deteriorated. Two children lost follow-up. We suggest that Pang's Classification of SCMs may be useful in describing pathological changes and guiding surgical procedure; imaging examine (including MRI, CT and X-ray) would play a significant role in confirmed SCMs diagnosis; and surgical operation should focus on eliminate and prevent spinal cord damnification.