Diabetic Angiopathies Clinical Trial
Official title:
Polymorphism Analysis of HLA-DRB1*04 Alleles in Patients With Type 2 Diabetic Macroangiopathy in Northeast Chinese
To explore the possible implications of HLA-DRB1*04 alleles in patients with type 2 DM and macroangiopathy
Previous studies examining the molecular etiology of diabetes mellitus in China and abroad
have mainly focused on the relationship between HLA and type 1 diabetes mellitus (DM) (1-3).
Due to the complexity of type 2 DM heredity, few researchers have studied the correlation
between type 2 DM and HLA. In recent years, it has been suggested that the development of
coronary heart disease (CHD) in diabetic individuals is not merely a complication of
diabetic macroangiopathy. CHD and diabetic macroangiopathy share some genetic associations.
Previous studies have demonstrated that certain HLA*DRB1*04 subtypes are associated with an
increased risk of cardiovascular disease.
Experimental methods: For genomic DNA extraction, 3-5ml venous blood samples were collected
from all patients. Coagulation was prevented using EDTA.
The procedure for PCR amplification was as follows: denaturing for 100s at 96°C, annealing
for 60s at 63°C, 1 cycle; denaturing for 10s at 96°C, annealing for 60s at 63°C, 9 cycles;
denaturing for 10 s at 96°C, annealing for 30 s at 59°C; extending for 30 s at 72°C, 20
cycles; maintaining at 4°C. Analyzing PCR products stained by Ethidium Bromide on 2.5%
agarose gel electrophoresis. Bands were observed using an ultraviolet light and a biological
imaging system.
Statistical method: The degree of agreement with Hardy-Weinberg equilibrium was determined
using the χ2 test. A t-test was employed for comparison of CRP levels (M ± s) and a χ2 test
was performed to compare allelic frequencies. The relative risk rate (RR) was calculated as
(Number of patients with the gene×Number of people in control group without the gene)
divided by (Number of people in control group with the gene×Number of the patients without
the gene). PC was calculated using Fisher's method if χ2 > 3.84.
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Observational Model: Case Control, Time Perspective: Prospective
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