Gene Polymorphism Associated With Macroangiopathy in Type 2 Diabetes Patients

Diabetic Angiopathies Clinical Trial

Official title:

Polymorphism Analysis of HLA-DRB1*04 Alleles in Patients With Type 2 Diabetic Macroangiopathy in Northeast Chinese

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT02882945
• Other study ID #: HarbinMU_WHW_001
• Status: Completed
• Phase: N/A
• First received: August 25, 2016
• Last updated: August 25, 2016
• Start date: May 2015
• Est. completion date: September 2015

Study information

• Verified date: August 2016
• Source: First Affiliated Hospital of Harbin Medical University
• Contact: n/a
• Is FDA regulated: No
• Health authority: China: Ministry of Health
• Study type: Observational

Clinical Trial Summary

To explore the possible implications of HLA-DRB1*04 alleles in patients with type 2 DM and macroangiopathy

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 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.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 470
• Est. completion date: September 2015
• Est. primary completion date: August 2015
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: Both
• Age group: 20 Years to 60 Years

Eligibility

Inclusion Criteria:

• clinical CHD (such as angina pectoris, myocardial infarction) diagnosed by dynamic electrocardiogram and ultrasonic cardiogram;

• coronary atherosclerosis confirmed by coronary angiographic examination;

• cerebral infarction diagnosed by cerebral CT;

• common carotid artery intimal-media thickness (IMT) =1.2 mm measured by Doppler ultrasonic examination;

• extensive irregular stenosis of a lower extremity artery (diameter < 3mm) or segmentally obstructed.

Exclusion Criteria:

• age < 20 years

• age > 60 years

Study Design

Observational Model: Case Control, Time Perspective: Prospective

Related Conditions & MeSH terms

• Diabetic Angiopathies

Intervention

Other:
• Group A
150 healthy blood donors without a family history of diabetes mellitus were enrolled in the study.
• Group B
200 cases of type 2 DM without complications were enrolled in the study.
• 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.

Locations

Country	Name	City	State
n/a

Sponsors (1)

Lead Sponsor	Collaborator
First Affiliated Hospital of Harbin Medical University

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Test of allelic frequencies	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).	At recruitment	No
Secondary	Evaluation of serum CRP(C-reactive protein) levels	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).	At recruitment	No