Objective： Although the kinase insert domain-containing receptor （KDR） gene play an very important role in the metastasis of cancer and is also as one of the molecular targets used in cancer therapy, mutation in th...Objective： Although the kinase insert domain-containing receptor （KDR） gene play an very important role in the metastasis of cancer and is also as one of the molecular targets used in cancer therapy, mutation in the tyrosine kinase （TK） domain of the KDR gene has not been reported. Here we detected the mutations and polymorphisms in the TK domain of KDR gene in human lung cancer patients and to give the basic evidence and clue for cancer prevention and target therapy. Methods： The entire sequence of exons 21, 22, 23 and 27 （which contain the coding sequence of tyrosine phosphorylation） in the TK domain of KDR gene in the patients with lung cancer and control healthy individuals were assayed by PCR and DNA sequencing. We also analyzed one non-coding single nucleotide polymorphisms （SNPs） in the KDR gene. Results： No mutations were found in exon 22, 23 and 27. One heterozygous mutation of c.＋2837 in exon 21 was found at a frequency of 2.08% （2/96） in the patients with lung cancer and none were detected in the healthy control individuals. The mutation was from a G to a A resulting in substitution of arginine with histidine residue. Conclusion： Our data suggested that we should focus on the mutation or SNP in the other regions or the expression levels of KDR gene, and the function of c.＋2837 mutation of KDR .qene may be needed further study in the future.展开更多
Objective N-methyl-D-aspartate（NMDA）receptor has been indicated to be involved in the pathogenesis of Alzheimer’s disease（AD）.The NMDA receptor subunit 2b（NR2B）has attracted more attention due to its characteri...Objective N-methyl-D-aspartate（NMDA）receptor has been indicated to be involved in the pathogenesis of Alzheimer’s disease（AD）.The NMDA receptor subunit 2b（NR2B）has attracted more attention due to its characteristic distribution and selective reduction in AD brain.The present study aimed to explore the association between NMDA gene polymorphism and AD.Methods A total of 63 AD patients and 68 normal controls in Shanghai city were employed in this study.Genotype of C2664T variant（rs1806201）in the exon13 of GRIN2B gene was determined by gene sequencing. Results Among AD patients,15（23.6%）subjects were identified as C/C genotype,and 35（55.6%）were identified as C/T genotype.The left 13（20.6%）subjects were identified as T/T genotype.In normal controls,15（22.1%）subjects were identified as C/C genotype,39（57.4%）as C/T genotype and 14（20.6%）as T/T genotype.The distribution frequency of neither GRIN2B C2664T genotype（P=0.895）nor allele（P=0.790）was significantly different between AD patients and normal controls,even when the subjects were stratified by gender and age of disease onset in AD patients.Conclusion The results suggest that there is no relation between GRIN2B C2664T polymorphism and AD in Chinese Han population of Shanghai City.展开更多
The detection of single amino-acid variants （SAVs） usually depends on single-nucleotide polymorphisms （SNPs） database. Here, we describe a novel method that discovers SAVs at proteome level independent of SNPs dat...The detection of single amino-acid variants （SAVs） usually depends on single-nucleotide polymorphisms （SNPs） database. Here, we describe a novel method that discovers SAVs at proteome level independent of SNPs data. Using mass spectrometry-based de novo sequencing algorithm, peptide-candidates are identified and compared with theoretical protein database to generate SAVs under pairing strategy, which is followed by database re-searching to control false discovery rate. in human brain tissues, we can confidently identify known and novel protein variants with diverse origins. Combined with DNA/RNA sequencing, we verify SAVs derived from DNA mutations, RNA alternative splicing, and unknown post-transcriptional mechanisms. Furthermore, quantitative analysis in human brain tissues reveals several tissue-specific differential expressions of SAVs. This approach provides a novel access to high-throughput detection of protein variants, which may offer the potential for clinical biomarker discovery and mechanistic research.展开更多
基金Supported by the grants from the National Natural Science Foundation of China (No. 30772531)Guangdong Provincial Medical Science and Technology Research Foundation (No. B2006001)China Postdoctoral Science Foundation (No. 20060400212)
文摘Objective： Although the kinase insert domain-containing receptor （KDR） gene play an very important role in the metastasis of cancer and is also as one of the molecular targets used in cancer therapy, mutation in the tyrosine kinase （TK） domain of the KDR gene has not been reported. Here we detected the mutations and polymorphisms in the TK domain of KDR gene in human lung cancer patients and to give the basic evidence and clue for cancer prevention and target therapy. Methods： The entire sequence of exons 21, 22, 23 and 27 （which contain the coding sequence of tyrosine phosphorylation） in the TK domain of KDR gene in the patients with lung cancer and control healthy individuals were assayed by PCR and DNA sequencing. We also analyzed one non-coding single nucleotide polymorphisms （SNPs） in the KDR gene. Results： No mutations were found in exon 22, 23 and 27. One heterozygous mutation of c.＋2837 in exon 21 was found at a frequency of 2.08% （2/96） in the patients with lung cancer and none were detected in the healthy control individuals. The mutation was from a G to a A resulting in substitution of arginine with histidine residue. Conclusion： Our data suggested that we should focus on the mutation or SNP in the other regions or the expression levels of KDR gene, and the function of c.＋2837 mutation of KDR .qene may be needed further study in the future.
基金supported by the Key Project of Shanghai Science and Technology Committee(No.08411951100,10ZR1425800)the National Major Special Project of Science and Technology of Ministry of Science and Technology,China(No.2008ZX09312-014,2008ZX09312-003)
文摘Objective N-methyl-D-aspartate（NMDA）receptor has been indicated to be involved in the pathogenesis of Alzheimer’s disease（AD）.The NMDA receptor subunit 2b（NR2B）has attracted more attention due to its characteristic distribution and selective reduction in AD brain.The present study aimed to explore the association between NMDA gene polymorphism and AD.Methods A total of 63 AD patients and 68 normal controls in Shanghai city were employed in this study.Genotype of C2664T variant（rs1806201）in the exon13 of GRIN2B gene was determined by gene sequencing. Results Among AD patients,15（23.6%）subjects were identified as C/C genotype,and 35（55.6%）were identified as C/T genotype.The left 13（20.6%）subjects were identified as T/T genotype.In normal controls,15（22.1%）subjects were identified as C/C genotype,39（57.4%）as C/T genotype and 14（20.6%）as T/T genotype.The distribution frequency of neither GRIN2B C2664T genotype（P=0.895）nor allele（P=0.790）was significantly different between AD patients and normal controls,even when the subjects were stratified by gender and age of disease onset in AD patients.Conclusion The results suggest that there is no relation between GRIN2B C2664T polymorphism and AD in Chinese Han population of Shanghai City.
文摘The detection of single amino-acid variants （SAVs） usually depends on single-nucleotide polymorphisms （SNPs） database. Here, we describe a novel method that discovers SAVs at proteome level independent of SNPs data. Using mass spectrometry-based de novo sequencing algorithm, peptide-candidates are identified and compared with theoretical protein database to generate SAVs under pairing strategy, which is followed by database re-searching to control false discovery rate. in human brain tissues, we can confidently identify known and novel protein variants with diverse origins. Combined with DNA/RNA sequencing, we verify SAVs derived from DNA mutations, RNA alternative splicing, and unknown post-transcriptional mechanisms. Furthermore, quantitative analysis in human brain tissues reveals several tissue-specific differential expressions of SAVs. This approach provides a novel access to high-throughput detection of protein variants, which may offer the potential for clinical biomarker discovery and mechanistic research.
