Objective To reveal the real cause of onset and lay the foundation of preimplantation genetic diagnosis for the future, prenatal diagnosis were performed for high-risk fetuses who were diagnosed tentatively as severe ...
详细信息
Objective To reveal the real cause of onset and lay the foundation of preimplantation genetic diagnosis for the future, prenatal diagnosis were performed for high-risk fetuses who were diagnosed tentatively as severe micromelia TD-I type(or OI-II type, ACG-I, II type, SRPS-III type etc) by the antenatal ultrasonography. At the same time, analysing and discussing the high mutation incidence and the homozygous mutation mechanism of p.R248 C of TD-I type to reveal the underlying mechanism. Method DNA of the fetal specimens were prepared by the microscale fast extraction method. The sick fetuses were diagnosed rapidly by the ARMS/RE method established for the high incidence of p.R248 C of FGFR3 gene. For those that the p.R248 C test is negative, sequence analysis was performed on other sites by sequencing. For those that FGFR3 gene mutation is negative, the COL1 A1, COL1 A2, COL2 A1, DYNC2 H1, NEK1 and other related gene were detected using one or two generation sequencing technology, and a series of methods were used to identify the pathogenicity of novel mutations. Result A total of 15 patients with severe short limb deformities were examined.(1) 10 cases carried the mutation of FGFR3 gene, thereinto, 6 cases with p.R248 C mutation, 1 case with p.S249 C mutation, 2 cases with p.Y373 C mutation, 1 case with a novel heterozygous mutation. For p.R248 C mutation, 3 homozygous mutation cases and 3 heterozygous ones were found. Using the ARMS/RE method, all normal control was negative, however, 365 bp specific fragments were amplfied for patients with p.R248 C mutation. And the fragment can be cut into two fragments(22 bp and 343 bp) by specific enzyme digestion.(2) 3 cases carried the mutation of COL1 A1 gene, case 1 and case 2 are twins who carried the p.G452 R mutation and case 3 carried the p.G1022 V mutation.(3) 1 case who carried the mutation of PPIB gene is compound heterozygous.(4) 1 case who carried the mutation of DYNC2 H1 gene is compound heterozygous. Conclusion
Objective Background and Objectives The precise level of NR0 B1 gene expression is critical f or gonadal differentiation during embryogenesis, and the ectopic activation of NR0 B1 is involved i n the development of so...
详细信息
Objective Background and Objectives The precise level of NR0 B1 gene expression is critical f or gonadal differentiation during embryogenesis, and the ectopic activation of NR0 B1 is involved i n the development of some cancers. However, the regulatory mechanisms controlling NR0 B1 exp ression are not well understood. Therefore, the epigenetic modifications promoting NR0 B1 activat ion were examined in this study. Method Methods The expression profile of NR0 B1 protein was investigated in 160 human lung a denocarcinoma(ADCA) cases by immunohistochemical analysis. The methylation pattern of the Cp G islands(CGI) of the NR0 B1 promoter was investigat ed in cancerous and paired adjacent noncancerous tissues by bisulfite sequencing. DNA methyltr ansferase inhibitor 5′-aza-2'-deoxycytidine, histone deacetylase inhibitor trichostatin A(TSA) and histone methylase inhibitors DZNep and BIX01294 were used to treat cells for up-regulating NR0 B1 transcription. The levels of modified-histones H3 and H4 occu pied on the NR0 B1 promoter were examined in the tissues and cells with different NR0 B1 expres sion status by the chromatin immunoprecipitation and quantitative PCR analysis. Result Results NR0 B1 protein was detected in cancerous tissues of more than 50% of ADCA ca ses and tended to be expressed in low-differentiated cancerous tissues from males. Nevertheless, NR0 B1 activation in ADCA has not previously been correlated with DNA demethylation. DNA methylation was not observed to be involved in the in vivo NR0 B1 transcription regulation althoug h the hypermethylated NR0 B1 CGI silenced its in vitro expression. NR0 B1 expression was not de tected in the 293 T cells although they possess a hypomethylated NR0 B1 promoter. The following TSA treatment increased acetylated histone H4 binding to the NR0 B1 promoter and activated NR0 B1 expression;in contrast, the treatment of DZNep and BIX01294 decreased the methylation of histones H3 K9 and H3 K27 and slightly induced NR0 B1 tra
Objective To reveal molecular genetic mechanism of a couple with normal phenotype but induce labor a thoracostenosis and phocomelia fetus. So that we can make good foundations of PGD for future high risk fetus. Method...
详细信息
Objective To reveal molecular genetic mechanism of a couple with normal phenotype but induce labor a thoracostenosis and phocomelia fetus. So that we can make good foundations of PGD for future high risk fetus. Method Because of no the 1 st aborted fetus‘ sample, we use parents‘ DNA to do whole-exome s equencing. Then the results were analyzed according to inheritance patterns, ultrasound results o f fetus, clinical symptoms, and more than forty common gene of bone diseases to select some ca ndidate mutations that we use Sanger sequence to verify them. Novel mutations were evaluated f or their potential pathogenecity using the SIFT, Poly Phen-2 algorithms, Provean and conservativ e analysis among more than 10 species. On the basic of determined etiology, we collected the a mniotic fluid from the 18 th gestational week. Amplification of relevant exons by PCR and bi-directi onal sequence. Result(1) Candidate gene from father: LEPRE1(c.1057 A>G), FLNB(c.3583 G>A), EVC(c.884 C>G), GUSB(c.10 G>A), ARSE(c.775 C >G), DYNC2 H1(p.F3571 Rfs*5, GALNS(c.1462 G>A), PCNT(c.5582 C>T), FLNA(c.1582 G>A), etc., total 19. From mother: AGL(c.686 A>G), SLC34 A1(c.533 T>A), ESCO2(c.1522 A>G), COL 5 A1(c.378 G>T), SLC34 A3(c.1612 C>T), DYNC2 H1(p.R2771 S), F BN1(c.3463+3 A>G), GDF5(c.979 C>G), etc., total 28.(2)After Sanger sequence and identificatio n of pathogenicity for the novel mutations, both F3571 Rfs*5 and R2771 S of DYNC2 H1 gene ar e pathogenic. Where the mutations of amino acids are highly conserved, the spatial conformation change caused by the mutations is significant. Conclusion(1)After NGS, Sanger sequence and identification of patho-genicity, the fetus was di agnosed with rare asphyxiating thoracic dysplasia type 3(ATD3), so termination of pregnancy wa s recommended. The root cause of this fetus‘ disease are F3571 Rfs*5 and R2771 S of DYNC2 H1 gene.(2)For rare genetic skeletal disorders, it can be detected with NGS and Sanger sequence.
Objective To investigate the association of functional variations(coding and UTR) of GWAS repo rted loci and exudative age-related macular degeneration(wet AMD) in Chinese *** Sixty-one genes in the 28 previous report...
详细信息
Objective To investigate the association of functional variations(coding and UTR) of GWAS repo rted loci and exudative age-related macular degeneration(wet AMD) in Chinese *** Sixty-one genes in the 28 previous reported GWAS loci were designed for target sequen cing of 400 wet AMD patients(200 PCV and 200 CNV) and 1250 controls. After single SNP asso ciation analysis, 25 variants in 18 of the 61 genes with association P value <0.01 were selected fo r the first replication in 885 wet AMDs(523 PCV and 362 CNV) and 562 controls by Sequenom M ass Array genotyping. After that, 4 SNPs were selected for further validation in the third Chinese c ohort with 456 wet AMDs(241 PCV and 215 CNV) and 211 controls by SNa Pshot genotyping. Result Previously reported six coding variants(rs800292, rs2274700, rs3753396 and rs1065489) in CFH and rs10490924 and rs2736911 in the HTRA1/ARMS2 region showed strong association with wet AMD. Previously reported four SNPs in loci 3 q12.1, 9 q31.1 and 15 q21.3 only showed ve ry week association in this Chinese cohort. The variant rs189132250(c.A1329 G, p.A443 A) in the BBX gene located in 3 q12.1 showed association with wet AMD with P value of 8.15? 10-5 and O R of 5.46 with stronger association in CNV than in PCV(P =7.06?10-6 and OR of 9.34 for CNV;P=0.021, OR=3.83 for PCV). rs144351944 in the UTR5 region of FILIP1 L gene showed associatio n only with PCV but not with CNV(P=9.06 ?10-4, OR=1.59 for PCV;P=0.30, OR=1.2 for CNV;P=2.13?10-3 and OR of 4.47 for wet AMD). rs1800978 in the UTR5 of ABCA1 also association wit h wet AMD mainly contributed from PCV but not CNV(P=1.17× 10?3, OR=1.26 for wet AMD;P=3.0× 10?4, OR=1.33 for PCV and P=0.04, OR=1.20 for CNV). Conclusion rs189132250 in BBX, rs144351944 in FILIP1 L and rs1800978 in the UTR5 of ABCA1 showed association in wet AMD. rs189132250 in BBX showed association mainly in CNV subty pe, while rs144351944 in FILIP1 Land rs1800978 in ABCA1 showed association mainly in PCV su btype. Th
暂无评论