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Trial record 2 of 16 for:    HLA-DRB1 AND DRB1

Gene Polymorphism Associated With Macroangiopathy in Type 2 Diabetes Patients

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ClinicalTrials.gov Identifier: NCT02882945
Recruitment Status : Completed
First Posted : August 30, 2016
Last Update Posted : August 30, 2016
Sponsor:
Information provided by (Responsible Party):
Weihua Wu, First Affiliated Hospital of Harbin Medical University

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
Diabetic Angiopathies Other: Group A Other: Group B Other: Group C

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 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.


Study Type : Observational
Actual Enrollment : 470 participants
Observational Model: Case Control
Time Perspective: Prospective
Official Title: Polymorphism Analysis of HLA-DRB1*04 Alleles in Patients With Type 2 Diabetic Macroangiopathy in Northeast Chinese
Study Start Date : May 2015
Actual Primary Completion Date : August 2015
Actual Study Completion Date : September 2015

Resource links provided by the National Library of Medicine


Group/Cohort 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 Name: healthy blood donors

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 Name: type 2 DM without complications

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 Name: type 2 DM with macroangiopathy complication




Primary Outcome Measures :
  1. Test of allelic frequencies [ Time Frame: 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 :
  1. Evaluation of serum CRP(C-reactive protein) levels [ Time Frame: 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).


Biospecimen Retention:   Samples With DNA
Laboratory analysis Blood samples were collected after over-night fasting, and serum was stored at −20°C.


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Ages Eligible for Study:   20 Years to 60 Years   (Adult)
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   Yes
Sampling Method:   Probability Sample
Study Population
150 healthy blood donors without a family history of diabetes mellitus at the Harbin Blood Disease Research Center were enrolled in the study, and 320 outpatients and hospitalized patients of type 2 diabetes mellitus from the First and Second Affiliated Hospital of Harbin Medical University.
Criteria

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

Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT02882945


Sponsors and Collaborators
First Affiliated Hospital of Harbin Medical University
Investigators
Principal Investigator: Nan Ma, Master First Affiliated Hospital of Harbin Medical University

Responsible Party: Weihua Wu, Director, First Affiliated Hospital of Harbin Medical University
ClinicalTrials.gov Identifier: NCT02882945     History of Changes
Other Study ID Numbers: HarbinMU_WHW_001
First Posted: August 30, 2016    Key Record Dates
Last Update Posted: August 30, 2016
Last Verified: August 2016
Individual Participant Data (IPD) Sharing Statement:
Plan to Share IPD: Undecided

Additional relevant MeSH terms:
Diabetic Angiopathies
Vascular Diseases
Cardiovascular Diseases
Diabetes Complications
Diabetes Mellitus
Endocrine System Diseases