| Objective: (1) To study the gene mutation in a pedigree with Dowling-Meara type epidermolysis bullosa simplex(DW-EBS) of monogenic disease. (2) To study the relationship between paraoxonasel(PON1) and coronary heart disease(CHD) of polygenic disease.Methods: (1) Using polymerase chain reaction(PCR) combined with single strand conformation polymorphism (SSCP) method and DNA sequencing to detect the point mutation of keratin 14 in a pedigree of DM-EBS. (2) The genotype and allele frequency of paraoxonasel gene -108C/T,-162A/G and -909C/G polymorphism were assayed by polymerase chain reaction(PCR)- restriction fragment length polymorphism(RFLP).Results: (1) Many bases were divergent between 637bp PCR product of the case and the codes of the same place of Genebank.(2) 121 code of K14 was CAC in the case in 151bp PCR product. That was different from AAC at the same place of Genebank. 123 code of K14 was GGT. That was different from AAT at the same place of Genebank,too. Other codes were same as the identical place of Genebank.(3) 121 code of K14 was CAC in the case and two controls in 151bp PCR products. 123 code was ACT in the case. 123 code was GGT in two controls. All of them were different from the codes of the same place of Genebank. Other codes were same as the identical place of Genebank.(4) Two PCR products were received from one sample. 121 code and 123 code of K14 were identical or not in two PCR products. Other codes were same as the identical places of Genebank.(5) PON1-108C/T:The frequencies of the genotypes and alleles in CHD were 0.105,0.547,0.349 and 0.378,0.622 respectively.The frequencies in control were 0.250,0.453,0.297 and 0.477,0.523 respectively.There was significant differences in genotype frequency between CHD group and control.PONl-162A/G: The frequencies of the genotypes and alleles in CHD were 0.035,0.105,0.860 and 0.087,0.913 respectively.The frequencies in control were 0.047,0.203,0.750 and 0.148,0.852 respectively.There was no significant differences in the genotype and allel frequency between CHD group and control.PONl-909C/G:The frequencies of the genotypes .andalleles in CHD were 0.233,0.558,0.209 and 0.512,0.488 respectively.The frequencies in control were 0.258,0.523,0.219 and 0.520,0.480 respectively. There was no significant differences in the genotype and allel frequency between CHD group and control,too.(6) The genotype of PON1-108TT was a risk factor of CHD (OR=2.807,P=0.022). The genotype of PON1-162GG and PON1-909GG rise the risk of CHD.But the ORs of them were no significant difference.(7) PON1-108C/T,-162A/G and -909C/G were combined to analysize the association with CHD.The OR with one risk genotype was 1.812,P=0.244. The OR with two risk genotypes was 1.636,P=0.360. The OR with three risk genotypes was 3.556,P=0.079.There were no significant differences.That was to say increasing number of risk genotypes was not associated with CHD.(8)The level of HDL-C and ApoA I were lower significantly in CHD group than in control(P<0.05),but the level of TG,TC and LDL-C were no significant difference between two groups(P>0.05). PONl-108C/T:The level of HDL-C in CC homozygotes was lower significantly in CHD group than in control(P<0.05),there was no significant difference in TT homozygotes and TC heterozygotes between two groups(P>0.05).The level of plasm lipids in two groups was no significant difference between PON1-162A/G and PON1-909C/G..Conclusion: (1) There is gene polymorphism at 121 code and 123 code of K14 possibly.(2) There is not gene mutation in 151bp gene fragment of K14 1A domain in the pedigree.(3)The -108C/T polymorphism of PON1 maybe associate with CHD in Chinese population.(4) The -162A/G and -909C/G polymorphism of PON1 are not associated with CHD in Chinese population.(5) PON1-108TT genotype maybe is a risk factor of CHD in Chinese.(6) There is no cooperative effect among these risk genotypes with CHD.
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