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Carriage of the TT-genotype, T-allele of the AGT gene, C-allele of the AT1R gene, 4a-allele in the 4a/4a+4a/4b group of ecNOS gene and the +9/+9- genotype of the B2BKR gene showed a significant higher level of FMD disturbances.
al., 2003), showed an association between the ecNOS gene and both endothelial function and hypertension however a number of similar studies were negative (Zee et.
Many studies have explored the association between ecNOS polymorphisms and vascular diseases (14).
A recent report discussed the relationship between the ecNOS polymorphism and gene expression and/or enzyme activity (16), but because the study was performed on postpartum placental tissues, the associations arrived at might have been modified by a history of maternal smoking and other environmental factors.
ecNOS is relatively unstable; we therefore harvested the cultured cells quickly and froze the harvested cells at -70[degrees]C before homogenization for mRNA measurement and Western blot analysis.
DNA ISOLATION AND IDENTIFICATION OF ecNOS POLYMORPHISMS
Analyses of the intron 4 variable number of tandem repeats (VNTR) polymorphism, the E298A polymorphism in exon 7, and the G10-T polymorphism in intron 23 of the ecNOS gene were performed as we have previously described (15).
Studies have rarely been performed on the specific association between the genetic polymorphisms of the ecNOS gene and plasma NOx concentrations in CAD patients.
The intron 4 VNTR polymorphism of the ecNOS gene was detected by the method of Wang et al.
For detection of the E298A polymorphism in exon 7 of the ecNOS gene, a primer pair was used to amplify a part of the ecNOS gene containing exon 7 by PCR.
For the G10-T polymorphism in intron 23 of the ecNOS gene, the primer pair for PCR was as follows: sense, 5'-CCCCTGAGTCATCTAAGTATTC-3'; antisense, 5'AGCTCTGGCACAGTCAAG-3'.
The G-allele frequency of the G10-T polymorphism in intron 23 of the ecNOS gene was higher in CAD patients than in controls.
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