·AIM: To investigate the clinical features and genetic defects in four generations of a Chinese family affected with atypical granular corneal dystrophy type I (GCD type I). · METHODS: Family history and cli...·AIM: To investigate the clinical features and genetic defects in four generations of a Chinese family affected with atypical granular corneal dystrophy type I (GCD type I). · METHODS: Family history and clinical data were recorded. Genomic DNA samples were obtained from peripheral blood leukocytes of all participated. Exons of the transforming growth factor-β-induced (TGFBI) gene were directly sequenced after being amplified by polymerase chain reaction (PCR), and multi-point linkage analysis using microsatellite makers flanking the gene was applied to identify the disease-causing mutation. · RESULTS: Clinical features were quite variable in patients, some patients only had opacities in the epithelium, and others revealed multiple bilateral circular, discrete, crumb -like opacities mainly in the epithelium, with several in different depths of corneal stroma, and the performance was different bilaterally, even in the same patient. Directly nucleotide sequencing revealed a heterozygous p.R555W mutation in the coding sequence of the TGFBI gene in all affected individuals of the family, but was not found in all unaffected. The maximum logarithm of odds (LOD) score obtained by multi -point analysis was detected at marker locus D5S393 (LOD = 2.740; α=1.000). ·CONCLUSION: Our case presented with clinical futures and the pathogenic mutations in TGFBI gene, the phenotype of the pedigree was quite different from typical GCD type I, so we suggested that this phenotype was a variant of GCD type I. These findings expand the knowledge about GCD type I, and demonstrate that molecular genetic analysis is important to make an accurate diagnosis of patients with variable corneal dystrophies in clinic.展开更多
In this paper,we report the clinical and molecular features of the distinct TGFBI (human transforming growth factor β-induced,OMIM No.601692) gene-linked corneal dystrophy.Altogether,five pedigrees and ten unrelated ...In this paper,we report the clinical and molecular features of the distinct TGFBI (human transforming growth factor β-induced,OMIM No.601692) gene-linked corneal dystrophy.Altogether,five pedigrees and ten unrelated individuals diagnosed as corneal dystrophy were recruited.Peripheral venous DNA was extracted,and then amplified by polymerase chain reaction (PCR) and scanned for mutation by single-stranded conformation polymorphism (SSCP).Direct DNA sequencing was used to analyze the mutations of the TGFBI gene.In our study,thirty patients from five pedigrees and ten sporadic patients were diagnosed as four TGFBI gene-linked corneal dystrophies of granular corneal dystrophy type I (GGCD I),Avellino corneal dystrophy (ACD),lattice corneal dystrophy type I (LCD I),and lattice corneal dystrophy type ⅢA (LCD IIIA),and in total,seven disease-causing mutations,namely R555W,A546D,A546T,and T538P mutations in exon 12,R124H and R124C mutations in exon 4,and P501T mutation in exon 11,were identified,while four polymorphisms of V327V,L472L,F540F,and 1665-1666insC were screened in exons 8,11,and 12.The study ascertained the tight genotype-phenotype relationship and confirmed the clinical and genetic features of four TGFBI gene-linked corneal dystrophies.展开更多
AIM: To analyze mutations in transforming growth factor beta-induced (TGFBI) gene in a Chinese pedigree with Reis-Bücklers corneal dystrophy (RBCD,also known as GCD3).METHODS: In a five-generation Chinese family,...AIM: To analyze mutations in transforming growth factor beta-induced (TGFBI) gene in a Chinese pedigree with Reis-Bücklers corneal dystrophy (RBCD,also known as GCD3).METHODS: In a five-generation Chinese family,eight members were identified with RBCD and the rest were unaffected.All members of the family underwent complete ophthalmologic examinations.Exons of TGFBI were amplified by polymerase chain reaction,sequenced,and compared with a reference database.RESULTS: A single heterozygous C>T (R124C) point mutation was found in exon 4 of TGFBI in all the affected members of the pedigree,but not in the unaffected members.CONCLUSION: R124C which was a known mutation for lattice corneal dystrophy type I,segregated with the RBCD in this pedigree.This elucidated the correlation between genotype and phenotype in a Chinese family of RBCD.展开更多
Background Both in vitro and in vivo data have demonstrated the TGFBI gene functions as a putative tumor suppressor and is frequently downregulated in human tumors of different histological types.The hypermethylation ...Background Both in vitro and in vivo data have demonstrated the TGFBI gene functions as a putative tumor suppressor and is frequently downregulated in human tumors of different histological types.The hypermethylation of the TGFBI promoter,as one of the main regulatory mechanisms,is associated with TGFBI silencing.In this study,we used a methylation-specific PCR （MSP） method to evaluate the methylation status of the TGFBI promoter in human leukemias.Methods Real-time RT-PCR and methylation-specific PCR approaches were performed to define the TGFBI expression and promoter methylation in human leukemia call lines and clinical samples.Genomic DNA was isolated from peripheral blood mononuclear cells from leukemia patients,bisulfite-converted,and analyzed by the MSP method.Results Hypermethylation of the TGFBI promoter occurred in leukemia cell lines and demethylation treatment reexpressed TGFBI at a substantially increased level in most of leukemia cell lines tested.Furthermore,a much higher level of CpG island methylation and a significantly lower TGFBI expression were also identified in clinical leukemia samples.Conclusion The results suggest an important role of promoter methylation in regulating TGFBI expression in leukemia,which provides a useful diagnostic marker for clinical management of human leukemias.展开更多
AIM:To investigate whether mutations in TGFBI gene or CHST6 gene correlated with stromal corneal dystrophies(CD) in 8 Chinese probands.· METHODS:Eight unrelated patients with stromal corneal dystrophies were recr...AIM:To investigate whether mutations in TGFBI gene or CHST6 gene correlated with stromal corneal dystrophies(CD) in 8 Chinese probands.· METHODS:Eight unrelated patients with stromal corneal dystrophies were recruited in this study;all affected members were assessed by completely ophthalmologic examinations.Genomic DNA was extracted from peripheral leukocytes,17 exons of TGFBI gene and the exon of CHST6 gene were amplified by polymerase chain reaction(PCR),sequenced directly and compared with the reference database.· RESULTS:Three heterozygous mutations in TGFBI gene were identified in six patients:c.370C>T(p.Arg124Cys) was found in exon 4 of TGFBI gene in three members,c.371G>A(p.Arg124His) was found in one patient;c.1663C>T(p.Arg555Trp) was found in exon 12 in other two members.In addition,four polymorphisms with the nucleotide changes rs1442,rs1054124,rs4669,and rs35151677 were found in TGFBI gene.Mutations were not identified in the rest of 2 affected individuals in TGFBI gene or CHST6 gene.· CONCLUSION:Within these patients,R124C,R124H and R555W mutations were co-segregated with the disease phenotypes and were specific mutations for lattice corneal dystrophy type I(LCD I),Avellino corneal dystrophy(ACD,GCDⅡ),granular corneal dystrophy type I(GCD I),respectively.Our study highlights the prevalence of codon 124 and codon 555 mutations in the TGFBI gene among the Chinese stromal corneal dystrophies patients.·展开更多
目的:探讨转化生长因子β诱导蛋白(TGFBI)在胶质瘤中的表达及其生物学功能。方法:通过从癌症基因组图谱(TCGA)、中国脑胶质瘤图谱(CGGA)和脑肿瘤分子数据库(Rembrandt)下载胶质瘤的转录组数据和相应的临床资料,分析在不同级别胶质瘤中TG...目的:探讨转化生长因子β诱导蛋白(TGFBI)在胶质瘤中的表达及其生物学功能。方法:通过从癌症基因组图谱(TCGA)、中国脑胶质瘤图谱(CGGA)和脑肿瘤分子数据库(Rembrandt)下载胶质瘤的转录组数据和相应的临床资料,分析在不同级别胶质瘤中TGFBI mRNA的表达水平,及其表达变化与胶质瘤患者预后的关系。免疫组织化学染色法检测35例胶质母细胞瘤组织和5例正常脑组织中TGFBI蛋白的表达水平。通过用慢病毒转染U87和U373细胞株构建TGFBI表达下调的胶质母细胞瘤细胞系,检测TGFBI对胶质瘤细胞增殖、侵袭、凋亡和细胞周期的影响。通过基因集富集分析(GSEA)预测TGFBI在胶质瘤发生发展中可能参与并调控的相关通路。结果:TCGA、CGGA和Rembrandt数据库的结果分析显示随着胶质瘤级别的增加,TGFBI mRNA的表达水平升高(P<0.05)。此外,TGFBI m RNA的高表达与胶质瘤患者不良预后显著相关(P<0.01)。免疫组化检测结果显示,与正常脑组织相比,胶质母细胞瘤组织中TGFBI蛋白表达水平显著增加(P<0.05)。细胞学实验结果显示,TGFBI表达下调可显著抑制胶质母细胞瘤细胞增殖和侵袭能力,同时能够促进细胞凋亡,促使细胞从G1期转向S期。GSEA分析结果显示高表达TGFBI可能通过TOLL样受体信号通路、NOD样受体信号通路和JAK-STAT信号通路参与调控胶质瘤恶性生物学行为。结论:TGFBI在胶质母细胞瘤组织中表达升高,可促进肿瘤细胞的增殖、侵袭,抑制细胞凋亡并诱导细胞周期重排,推测其可能在胶质瘤发生发展中扮演着癌基因的角色。展开更多
基金Zhejiang Key Innovation Team Project of China (No.2009R50039)Zhejiang Key Laboratory Found of China (No.2011E10006)+1 种基金Medical Science and Technology Project of Zhejiang Province,China (No.2010QNA012)Science and Technology Program of Zhejiang University (No.2011FZA7013)
文摘·AIM: To investigate the clinical features and genetic defects in four generations of a Chinese family affected with atypical granular corneal dystrophy type I (GCD type I). · METHODS: Family history and clinical data were recorded. Genomic DNA samples were obtained from peripheral blood leukocytes of all participated. Exons of the transforming growth factor-β-induced (TGFBI) gene were directly sequenced after being amplified by polymerase chain reaction (PCR), and multi-point linkage analysis using microsatellite makers flanking the gene was applied to identify the disease-causing mutation. · RESULTS: Clinical features were quite variable in patients, some patients only had opacities in the epithelium, and others revealed multiple bilateral circular, discrete, crumb -like opacities mainly in the epithelium, with several in different depths of corneal stroma, and the performance was different bilaterally, even in the same patient. Directly nucleotide sequencing revealed a heterozygous p.R555W mutation in the coding sequence of the TGFBI gene in all affected individuals of the family, but was not found in all unaffected. The maximum logarithm of odds (LOD) score obtained by multi -point analysis was detected at marker locus D5S393 (LOD = 2.740; α=1.000). ·CONCLUSION: Our case presented with clinical futures and the pathogenic mutations in TGFBI gene, the phenotype of the pedigree was quite different from typical GCD type I, so we suggested that this phenotype was a variant of GCD type I. These findings expand the knowledge about GCD type I, and demonstrate that molecular genetic analysis is important to make an accurate diagnosis of patients with variable corneal dystrophies in clinic.
基金Project supported by the Ministry of Health Research Fund of China(No. WKJ2009-2-020)the Science and Technology Specific Project of Zhejiang Province (No. 2009C03010-2),China
文摘In this paper,we report the clinical and molecular features of the distinct TGFBI (human transforming growth factor β-induced,OMIM No.601692) gene-linked corneal dystrophy.Altogether,five pedigrees and ten unrelated individuals diagnosed as corneal dystrophy were recruited.Peripheral venous DNA was extracted,and then amplified by polymerase chain reaction (PCR) and scanned for mutation by single-stranded conformation polymorphism (SSCP).Direct DNA sequencing was used to analyze the mutations of the TGFBI gene.In our study,thirty patients from five pedigrees and ten sporadic patients were diagnosed as four TGFBI gene-linked corneal dystrophies of granular corneal dystrophy type I (GGCD I),Avellino corneal dystrophy (ACD),lattice corneal dystrophy type I (LCD I),and lattice corneal dystrophy type ⅢA (LCD IIIA),and in total,seven disease-causing mutations,namely R555W,A546D,A546T,and T538P mutations in exon 12,R124H and R124C mutations in exon 4,and P501T mutation in exon 11,were identified,while four polymorphisms of V327V,L472L,F540F,and 1665-1666insC were screened in exons 8,11,and 12.The study ascertained the tight genotype-phenotype relationship and confirmed the clinical and genetic features of four TGFBI gene-linked corneal dystrophies.
基金Supported by grants from the National Natural Science Foundation of China (No. NNSF 81000370)
文摘AIM: To analyze mutations in transforming growth factor beta-induced (TGFBI) gene in a Chinese pedigree with Reis-Bücklers corneal dystrophy (RBCD,also known as GCD3).METHODS: In a five-generation Chinese family,eight members were identified with RBCD and the rest were unaffected.All members of the family underwent complete ophthalmologic examinations.Exons of TGFBI were amplified by polymerase chain reaction,sequenced,and compared with a reference database.RESULTS: A single heterozygous C>T (R124C) point mutation was found in exon 4 of TGFBI in all the affected members of the pedigree,but not in the unaffected members.CONCLUSION: R124C which was a known mutation for lattice corneal dystrophy type I,segregated with the RBCD in this pedigree.This elucidated the correlation between genotype and phenotype in a Chinese family of RBCD.
基金This study was supported by the National Basic Research Program of China （973 Program, No. 2013CB911000） and the National Nature Science Foundation of China （No. 30971602, 81272929）.Acknowledgements： We thank Dr. Chi Zhenfen for help with cell culture.
文摘Background Both in vitro and in vivo data have demonstrated the TGFBI gene functions as a putative tumor suppressor and is frequently downregulated in human tumors of different histological types.The hypermethylation of the TGFBI promoter,as one of the main regulatory mechanisms,is associated with TGFBI silencing.In this study,we used a methylation-specific PCR （MSP） method to evaluate the methylation status of the TGFBI promoter in human leukemias.Methods Real-time RT-PCR and methylation-specific PCR approaches were performed to define the TGFBI expression and promoter methylation in human leukemia call lines and clinical samples.Genomic DNA was isolated from peripheral blood mononuclear cells from leukemia patients,bisulfite-converted,and analyzed by the MSP method.Results Hypermethylation of the TGFBI promoter occurred in leukemia cell lines and demethylation treatment reexpressed TGFBI at a substantially increased level in most of leukemia cell lines tested.Furthermore,a much higher level of CpG island methylation and a significantly lower TGFBI expression were also identified in clinical leukemia samples.Conclusion The results suggest an important role of promoter methylation in regulating TGFBI expression in leukemia,which provides a useful diagnostic marker for clinical management of human leukemias.
文摘AIM:To investigate whether mutations in TGFBI gene or CHST6 gene correlated with stromal corneal dystrophies(CD) in 8 Chinese probands.· METHODS:Eight unrelated patients with stromal corneal dystrophies were recruited in this study;all affected members were assessed by completely ophthalmologic examinations.Genomic DNA was extracted from peripheral leukocytes,17 exons of TGFBI gene and the exon of CHST6 gene were amplified by polymerase chain reaction(PCR),sequenced directly and compared with the reference database.· RESULTS:Three heterozygous mutations in TGFBI gene were identified in six patients:c.370C>T(p.Arg124Cys) was found in exon 4 of TGFBI gene in three members,c.371G>A(p.Arg124His) was found in one patient;c.1663C>T(p.Arg555Trp) was found in exon 12 in other two members.In addition,four polymorphisms with the nucleotide changes rs1442,rs1054124,rs4669,and rs35151677 were found in TGFBI gene.Mutations were not identified in the rest of 2 affected individuals in TGFBI gene or CHST6 gene.· CONCLUSION:Within these patients,R124C,R124H and R555W mutations were co-segregated with the disease phenotypes and were specific mutations for lattice corneal dystrophy type I(LCD I),Avellino corneal dystrophy(ACD,GCDⅡ),granular corneal dystrophy type I(GCD I),respectively.Our study highlights the prevalence of codon 124 and codon 555 mutations in the TGFBI gene among the Chinese stromal corneal dystrophies patients.·
文摘目的:探讨转化生长因子β诱导蛋白(TGFBI)在胶质瘤中的表达及其生物学功能。方法:通过从癌症基因组图谱(TCGA)、中国脑胶质瘤图谱(CGGA)和脑肿瘤分子数据库(Rembrandt)下载胶质瘤的转录组数据和相应的临床资料,分析在不同级别胶质瘤中TGFBI mRNA的表达水平,及其表达变化与胶质瘤患者预后的关系。免疫组织化学染色法检测35例胶质母细胞瘤组织和5例正常脑组织中TGFBI蛋白的表达水平。通过用慢病毒转染U87和U373细胞株构建TGFBI表达下调的胶质母细胞瘤细胞系,检测TGFBI对胶质瘤细胞增殖、侵袭、凋亡和细胞周期的影响。通过基因集富集分析(GSEA)预测TGFBI在胶质瘤发生发展中可能参与并调控的相关通路。结果:TCGA、CGGA和Rembrandt数据库的结果分析显示随着胶质瘤级别的增加,TGFBI mRNA的表达水平升高(P<0.05)。此外,TGFBI m RNA的高表达与胶质瘤患者不良预后显著相关(P<0.01)。免疫组化检测结果显示,与正常脑组织相比,胶质母细胞瘤组织中TGFBI蛋白表达水平显著增加(P<0.05)。细胞学实验结果显示,TGFBI表达下调可显著抑制胶质母细胞瘤细胞增殖和侵袭能力,同时能够促进细胞凋亡,促使细胞从G1期转向S期。GSEA分析结果显示高表达TGFBI可能通过TOLL样受体信号通路、NOD样受体信号通路和JAK-STAT信号通路参与调控胶质瘤恶性生物学行为。结论:TGFBI在胶质母细胞瘤组织中表达升高,可促进肿瘤细胞的增殖、侵袭,抑制细胞凋亡并诱导细胞周期重排,推测其可能在胶质瘤发生发展中扮演着癌基因的角色。
