Gene Polymorphism Associated With Macroangiopathy in Type 2 Diabetes Patients
Study Details
Study Description
Brief Summary
To explore the possible implications of HLA-DRB1*04 alleles in patients with type 2 DM and macroangiopathy
| Condition or Disease | Intervention/Treatment | Phase |
|---|---|---|
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Detailed Description
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 HLADRB104 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.
Study Design
Arms and Interventions
| Arm | Intervention/Treatment |
|---|---|
| Group A 150 healthy blood donors without a family history of diabetes mellitus |
Other: Group A
150 healthy blood donors without a family history of diabetes mellitus were enrolled in the study.
Other Names:
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| Group B 200 cases of type 2 DM without complications (group B), there were 62 males and 108 females, aged 29-53, with an average age of 42 ± 9 years |
Other: Group B
200 cases of type 2 DM without complications were enrolled in the study.
Other Names:
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| Group C 120 cases of type 2 DM with macroangiopathy complication (group C), there were 70 males and 50 females, aged 40-53, with an average age of 47 ± 6 years. Among the group C subjects, 65 individuals had CHD; 55 patients had cerebrovascular disease (CVD); and 30 subjects had a combination of peripheral vascular diseases (PVD) and CHD |
Other: Group C
120 cases of type 2 DM with macroangiopathy complication were enrolled in the study.. Among the group C subjects, 65 individuals had CHD; 55 patients had cerebrovascular disease (CVD); and 30 subjects had a combination of peripheral vascular diseases (PVD) and CHD.
Other Names:
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Outcome Measures
Primary Outcome Measures
- Test of allelic frequencies [At recruitment]
A commercially available kit was used for extraction of genomic DNA from the whole blood samples (PEL-FREEZ Inc., USA). Thirty-two pairs of sequence-specific primers (SSP) for HLA-DRB1*04 alleles were purchased from One Lambda, Inc. (USA), and the Taq enzyme was purchased from Promega Corp. (USA). The PCR-SSP technique was employed to determine the HLA-DRB1*04 alleles for each subject. Genes were amplified using the 5700 PCR thermal cycler manufactured by Applied Biosystems Inc. (USA).
Secondary Outcome Measures
- Evaluation of serum CRP(C-reactive protein) levels [At recruitment]
A commercially available 96-well plate ELISA assay kit (introassay CV 1.7%~3.9%, interassay CV 2.8%~5.1%) was used to quantify hs-CRP in serum (DSL Inc., USA). All samples were evaluated in duplicate. The absorbance (OD) was determined using a BIO-RAD2550 microwell plate reader (USA).
Eligibility Criteria
Criteria
Inclusion Criteria:
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clinical CHD (such as angina pectoris, myocardial infarction) diagnosed by dynamic electrocardiogram and ultrasonic cardiogram;
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coronary atherosclerosis confirmed by coronary angiographic examination;
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cerebral infarction diagnosed by cerebral CT;
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common carotid artery intimal-media thickness (IMT) ≥1.2 mm measured by Doppler ultrasonic examination;
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extensive irregular stenosis of a lower extremity artery (diameter < 3mm) or segmentally obstructed.
Exclusion Criteria:
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age < 20 years
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age > 60 years
Contacts and Locations
Locations
No locations specified.Sponsors and Collaborators
- First Affiliated Hospital of Harbin Medical University
Investigators
- Principal Investigator: Nan Ma, Master, First Affiliated Hospital of Harbin Medical University
Study Documents (Full-Text)
None provided.More Information
Publications
None provided.- HarbinMU_WHW_001