RESUMO
Associations between polymorphisms of the CD36 gene and susceptibility to coronary artery heart disease (CHD) are not clear. We assessed allele frequencies and genotype distributions of CD36 gene polymorphisms in 112 CHD patients and 129 control patients using semi-quantitative polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. Additionally, we detected CD36 mRNA expression by real-time quantitative PCR, and we quantified plasma levels of oxidized low-density lipoprotein (ox-LDL) using an enzyme-linked immunosorbent assay (ELISA). There were no significant differences between the two groups (P>0.05) in allele frequencies of rs1761667 or in genotype distribution and allele frequencies of rs3173798. The genotype distribution of rs1761667 significantly differed between CHD patients and controls (P=0.034), with a significantly higher frequency of the AG genotype in the CHD group compared to the control group (P=0.011). The plasma levels of ox-LDL in patients with the AG genotype were remarkably higher than those with the GG and AA genotypes (P=0.010). In a randomized sample taken from patients in the two groups, the CD36 mRNA expression of the CHD patients was higher than that of the controls. In CHD patients, the CD36 mRNA expression in AG genotype patients was remarkably higher than in those with an AA genotype (P=0.005). After adjusted logistic regression analysis, the AG genotype of rs1761667 was associated with an increased risk of CHD (OR=2.337, 95% CI=1.336-4.087, P=0.003). In conclusion, the rs1761667 polymorphism may be closely associated with developing CHD in the Chongqing Han population of China, and an AG genotype may be a genetic susceptibility factor for CHD.
Assuntos
Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , /genética , Doença da Artéria Coronariana/genética , Predisposição Genética para Doença , Polimorfismo Genético/genética , Povo Asiático/genética , Estudos de Casos e Controles , China/etnologia , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/etnologia , Ensaio de Imunoadsorção Enzimática , Frequência do Gene , Genótipo , Predisposição Genética para Doença/etnologia , Modelos Logísticos , Lipoproteínas LDL/sangue , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição/genética , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , RNA Mensageiro/análiseRESUMO
OBJECTIVE: To investigate the functions of Fibroblast Growth Factor Receptor-2 (FGFR2) at different stages of cell differentiation. The engineered murine embryonic stem (ES) cells with conditional knockout of FGFR2 were developed depending on Cre-loxP. METHODS: Cre-loxP system was used in a conditional targeting vector. The competent AM-1 bacteria, which expressed Cre-recombinase, was used to confirm the Cre-mediated deletion of the floxed exons 7 and 8 of FGFR2. The targeting vector was electroporated into the ES cells, and the transfected ES cells were screened with G418 and Ganciclovir. Finally, the ES clones with correct targeting events were identified by Southern Blot and PCR. RESULTS: The targeting vector with conditional knockout of murine FGFR2 was successfully constructed andconfirmed by PCR and digesti on analysis in bacteria. 86 ES clones were collected by selective culture with G418 and Ganciclovir. Four of the 86 ES clones were found containing the targeting gene sequence in genomic DNA proved by Southern Blot with a 5'-end flank probe. Two of the four ES clones had the correct targeting events that included the insertion of the targeting gene sequence in genomic DNA and were checked by Southern Blot with a 3'-end flanking probe. Finally, the insertion of loxP (loxP3) between exons 8 and 9 in genomic DNA was identified in one of the two ES clones by Southern Blot and PCR.CONCLUSION: FGFR2 conditional knockout depending on Cre-loxP can be successfully used in ES cells.
Assuntos
Animais , Camundongos , Células-Tronco Embrionárias/metabolismo , Estruturas Embrionárias/citologia , Marcação de Genes , Genoma/genética , Vetores Genéticos/genética , Sequência de Bases , Integrases/genética , Integrases/metabolismo , Dados de Sequência Molecular , Recombinação Genética , Mapeamento por Restrição , Análise de Sequência de DNARESUMO
Sixteen adult patients (8 males and 8 females) with severe sickle cell painful crisis were given acupuncture for pain relief. Good response was achieved in 12 (75 per cent) patients and the remainder had moderate pain relief. All patients were discharged home the same day feeling better