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A 1-month-old male infant presented with skin flushing covering with collodion-like membrane all over the body at birth, and experienced gradual skin desquamation thereafter. At the age of 2 months, collodion-like membrane completely peeled off, and the patient presented with obvious scales and dry skin. Skin examination showed generalized dry skin, tense, glossy and transparent plastic wrapper-like membrane remaining on the front chest, large and disk-shaped white scales with an adherent center and free edges inlaid in the skin of the trunk and scalp. Genetic testing showed compound heterozygous mutations in the CYP4F22 gene of the patient, including the mutation c.1137G>A (p.W379X) inherited from his father and the mutation c.467G>A (p.R156H) inherited from his mother. The patient was diagnosed with lamellar ichthyosis.
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Objective@#To evaluate the application value and significance of low-depth whole-genome sequencing for copy number variations (CNV-Seq) in the genetic diagnosis and prenatal diagnosis of X-linked ichthyosis (XLI) due to STS gene deletion.@*Methods@#Clinical data were collected from 3 616 subjects who received CNV-Seq, and single-gene test results were collected from 7 patients or pedigrees with ichthyosis in The First Affiliated Hospital of Zhengzhou University in 2018. The 3 616 samples included 2 891 prenatal samples from pregnant women (most were amniotic fluid samples, some fetal villus samples, very few umbilical blood samples) and 725 peripheral blood samples from other subjects. Genomic DNA was extracted from amniocytes or peripheral blood, and then subjected to CNV-Seq. Quantitative PCR (qPCR) and single nucleotide polymorphism (SNP) -comparative genomic hybridization (CGH) array were performed to verify the detected CNVs. Pathogenicity of the CNVs was analyzed according to the database of genomic variants (DGV) , database of genomic variation and phenotype in humans using ensembl resources (DECIPHER) , clinical genome resource (ClinGen) and online Mendelian inheritance in man (OMIM) .@*Results@#Of the 3 616 subjects receiving CNV-Seq, Xp22.31 deletion was identified in prenatal samples from 6 pregnant women, including 5 male and 1 female fetuses. The deleted fragment of Xp22.31 covered the XLI region containing the major gene STS. The parental CNV-Seq showed that the Xp22.31 deletion was spontaneous mutation in 2 of the 6 fetuses, and inherited from the parents in the other 4 fetuses. qPCR confirmed that the female fetus was a carrier of a complete heterozygous deletion of the STS gene, and there was a complete deletion of the STS gene in the other 5 male fetuses. SNP-CGH array also confirmed that the female fetus was heterozygous Xp22.31 deletion carrier, which was consistent with the CNV-Seq results. Ichthyosis gene panel sequencing in the 7 patients with ichthyosis showed 1 with harlequin ichthyosis, 2 with ichthyosis vulgaris, 3 with XLI, and no causative mutation in 1. CNV-Seq confirmed that Xp22.31 deletion existed in the above 2 patients with XLI due to STS gene deletion. Moreover, Xp22.31 duplication was found in 16 out of 3 616 subjects receiving CNV-Seq, but they were all individuals or fetuses with normal phenotype.@*Conclusions@#CNV-Seq is a stable and reliable method for screening whole-genome CNVs, and can be applied to genetic diagnosis and prenatal diagnosis of XLI due to STS gene deletion. The deletion of Xp22.31 fragment containing the STS gene can cause XLI, and the duplication of the same region is highly likely to be the polymorphic variation.
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Objective To evaluate the application value and significance of low-depth whole-genome sequencing for copy number variations(CNV-Seq)in the genetic diagnosis and prenatal diagnosis of X-linked ichthyosis(XLI)due to STS gene deletion. Methods Clinical data were collected from 3616 subjects who received CNV-Seq, and single-gene test results were collected from 7 patients or pedigrees with ichthyosis in The First Affiliated Hospital of Zhengzhou University in 2018. The 3616 samples included 2891 prenatal samples from pregnant women(most were amniotic fluid samples, some fetal villus samples, very few umbilical blood samples)and 725 peripheral blood samples from other subjects. Genomic DNA was extracted from amniocytes or peripheral blood, and then subjected to CNV-Seq. Quantitative PCR(qPCR)and single nucleotide polymorphism(SNP)-comparative genomic hybridization(CGH)array were performed to verify the detected CNVs. Pathogenicity of the CNVs was analyzed according to the database of genomic variants(DGV), database of genomic variation and phenotype in humans using ensembl resources (DECIPHER), clinical genome resource (ClinGen) and online Mendelian inheritance in man (OMIM). Results Of the 3616 subjects receiving CNV-Seq, Xp22.31 deletion was identified in prenatal samples from 6 pregnant women, including 5 male and 1 female fetuses. The deleted fragment of Xp22.31 covered the XLI region containing the major gene STS. The parental CNV-Seq showed that the Xp22.31 deletion was spontaneous mutation in 2 of the 6 fetuses, and inherited from the parents in the other 4 fetuses. qPCR confirmed that the female fetus was a carrier of a complete heterozygous deletion of the STS gene, and there was a complete deletion of the STS gene in the other 5 male fetuses. SNP-CGH array also confirmed that the female fetus was heterozygous Xp22.31 deletion carrier, which was consistent with the CNV-Seq results. Ichthyosis gene panel sequencing in the 7 patients with ichthyosis showed 1 with harlequin ichthyosis, 2 with ichthyosis vulgaris, 3 with XLI, and no causative mutation in 1. CNV-Seq confirmed that Xp22.31 deletion existed in the above 2 patients with XLI due to STS gene deletion. Moreover, Xp22.31 duplication was found in 16 out of 3616 subjects receiving CNV-Seq, but they were all individuals or fetuses with normal phenotype. Conclusions CNV-Seq is a stable and reliable method for screening whole-genome CNVs, and can be applied to genetic diagnosis and prenatal diagnosis of XLI due to STS gene deletion. The deletion of Xp22.31 fragment containing the STS gene can cause XLI, and the duplication of the same region is highly likely to be the polymorphic variation.
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OBJECTIVE To explore whether different concentrations of potassium bisperoxo (1,10-phenanthroline) oxovanadate [BPV (phen)) affect cell cycle by regulating the expression of DNA meth-yltranferases (DNMT) and cell cycle-related genes. METHODS HT22 cells were treated with BPV (phen) 0.3 and 3 μ mol • L-1 for 24 h. The cell viability was detected by MTS assay, cell cycle was detected by flow cytometry, the activity of DNMT was detected by ELISA, the mRNA expressions of p21, DNMT1, DNMT3A and DNMT3B were measured with real-time quantitative PCR, while the levels of corresponding proteins were measured by Western blotting. RESULTS Compared with the DMSO control group, BPV(phen) 0.3 μ mol• L-1 did not affect cell viability, but the cell viability of BPV(phen) 3 pmol-L-1 increased signifi-cantly (P<0.05). There was no significant difference in cell cycle between DMSO control and BPV(phen) 0.3 nmol-L-1 group, but the proportion of cells in S phase increased((76.0±1.6)%)(P<0.05) and in G2 phase decreased ((2.1 ±1.5)%) (P<0.05) of BPV(phen) 3.0 μ mol • L-1 group. The intracellular DNMT activity of BPV(phen) 3.0 Mmol • L-1 group was significantly increased compared with the DMSO control group (F<0.05). There was no significant difference in the mRNA expressions of p21, DNMT1, DNMT3A and DNMT3B between DMSO control and BPV(phen) 0.3 μ mol • L-1 group, but all increased in BPV(phen) 3.0 μ mol • L-1 group (P<0.05, P<0.01). Western blotting results showed that there was no significant difference in the protein expressions of P21, DNMT1, DNMT3A and DNMT3B between DMSO control and BPV (phen) 0.3 μ mol • L-1 group, but only the protein expressions of DNMT3B and P21 of BPV (phen) 3.0 μ mol • L-1 group increased significantly (F<0.05). CONCLUSION BPV(phen) can regulate the expression of downstream and cell cycle-related genes by altering the expression of DNMTs, which in turn affects the growth and proliferation of HT22 cells.
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Abstract Objective: To identify pathogenic genomic imbalances in patients presenting congenital heart disease (CHD) with extra cardiac anomalies and exclusion of 22q11.2 deletion syndrome (22q11.2 DS). Methods: 78 patients negative for the 22q11.2 deletion, previously screened by fluorescence in situ hybridization (FISH) and/or multiplex ligation probe amplification (MLPA) were tested by chromosomal microarray analysis (CMA). Results: Clinically significant copy number variations (CNVs ≥300 kb) were identified in 10% (8/78) of cases. In addition, potentially relevant CNVs were detected in two cases (993 kb duplication in 15q21.1 and 706 kb duplication in 2p22.3). Genes inside the CNV regions found in this study, such as IRX4, BMPR1A, SORBS2, ID2, ROCK2, E2F6, GATA4, SOX7, SEMAD6D, FBN1, and LTPB1 are known to participate in cardiac development and could be candidate genes for CHD. Conclusion: These data showed that patients presenting CHD with extra cardiac anomalies and exclusion of 22q11.2 DS should be investigated by CMA. The present study emphasizes the possible role of CNVs in CHD.
Resumo Objetivo: Identificar desequilíbrios genômicos patogênicos em pacientes que apresentam cardiopatias congênitas (CC) e anomalias extracardíacas e exclusão da síndrome de deleção 22q11.2 (SD22q11.2). Métodos: Foram avaliados por microarray cromossômico (CMA) 78 pacientes negativos para a deleção 22q11.2, previamente testados por hibridação in situ com fluorescência (FISH) e/ou amplificação de múltiplas sondas dependentes de ligação (MLPA). Resultados: Foram identificadas variações do número de cópias de DNA (CNVs) clinicamente significativas (≥ 300 kb) em 10% (8/78) dos casos, além de CNVs potencialmente relevantes em dois casos (duplicação de 993 kb em 15q21.1 e duplicação de 706 kb em 2p22.3). Genes envolvidos como IRX4, BMPR1A, SORBS2, ID2, ROCK2, E2F6, GATA4, SOX7, SEMAD6D, FBN1 e LTPB1 são conhecidos por atuar no desenvolvimento cardíaco e podem ser genes candidatos a CC. Conclusão: Esses dados mostram que pacientes que apresentam CC, com anomalias extracardíacas e exclusão da SD22q11.2, devem ser investigados por CMA. Ainda, este estudo enfatiza a possível função das CNVs nas CC.
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Humans , Male , Female , Infant , Child , Adult , Chromosomes, Human, Pair 22/genetics , Chromosome Deletion , DNA Copy Number Variations/genetics , Heart Defects, Congenital/genetics , Oligonucleotide Array Sequence Analysis , GenomicsABSTRACT
Objective To investigate the genetic data of 21 autosom al STR included in G oldeneyeTM DNA ID 22N CK it in Chinese H an nationality and to evaluate the forensic application. Methods B y detected 500 unrelated healthy individuals in Chinese H an nationality of East China w ith G oldeneyeTM DNA ID 22N CK it, allele frequencies, population genetics param eters and linkage disequilibrium inform ation of the 21 autosom al STR w ere statistically analyzed. Results In the 21 autosom al STR , no deviations from H ardy-W einberg equilibrium w ere detected and all loci w ere independent form each other. D P values of 21 au-tosom al STR w ere all above 0.85, and the com bined discrim ination pow er w as 1-3.616 5×10-26. Com bined m ean exclusion chance of this system in duo cases w as 1-2.786 81×10-6, in trio cases w as 1-8.545 82× 10-10. Conclusion Tw enty-one autosom al STR included in G oldeneyeTM DNA ID 22N CK it are highly polym orphic in the H an nationality. Com bined w ith G oldeneyeTM DNA ID 20A K it, the kit can satisfy the needs for full-sibling testing and facilitate the solution of this kind of case tools.
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Epithelial membrane protein 3 (EMP3) is a trans-membrane signaling molecule with important roles in the regulation of apoptosis, differentiation and invasion of cancer cells, but the detailed is largely still unknown. We analyzed the mRNA levels and methylation statuses of EMP3 in 63 primary breast carcinomas and assessed their correlations with clinicopathologic variables. The expression of EMP3 mRNA in primary breast carcinomas was significantly higher than the expression of 20 normal breast tissues (p<10(-7)). EMP3 overexpression in breast carcinomas was significantly related to histological grade III (p=3.9X10(-7)), lymph node metastasis (p= 0.003), and strong Her-2 expression (p=3.3X10(-6)). Hypermethylation frequencies of EMP3 were detected in 36.5% of breast carcinomas by methylation-specific polymerase chain reaction. However, no significant correlations were found between methylation status of EMP3 and mRNA expression levels as well as other clinical parameters. In conclusion, EMP3 may be a novel marker of tumor aggressiveness. Overexpression of EMP3 in primary breast carcinoma is not associated with DNA methylation.
Subject(s)
Adult , Female , Humans , Middle Aged , Breast Neoplasms/genetics , Carcinoma/genetics , DNA Methylation , Epigenesis, Genetic , Gene Expression Regulation, Neoplastic , Lymphatic Metastasis , Membrane Glycoproteins/genetics , Neoplasm Staging , RNA, Messenger/metabolism , Receptor, ErbB-2/genetics , Severity of Illness IndexABSTRACT
AIM:To study the mRNA expression and methylation status of imprinted gene SLC22A18 in infiltrating ductal carcinomas(IDCs),and the correlation between methylation status and clinical characteristics in IDCs.METHODS:The methylation status at the promoter regions of SLC22A18 gene was examined by methylation-specific polymerase chain reaction(MSP)in the specimens of IDC from 40 patients.The mRNA expression of SLC22A18 gene was detected by real-time reverse quantitative transcriptase polymerase chain reaction(real-time RT-PCR)in 40 specimens of IDC and the cell line MDA-MB-231.The cell line MDA-MB-231 was treated with 5-aza-2'-deoxycytidine(5'-aza-dc)and trichostatin A(TSA),then MSP and rea1-time RT-PCR were used to detect the methylation status and mRNA expression levels of SLC22A18 gene.RESULTS:SLC22A18 mRNA expression in 40 IDC tissues was lower than that in al1 corresponding adjacent non-cancerous tissues(P