Biomarker for diabetes mellitus complicated with coronary heart disease

A technology for coronary heart disease and diabetes, applied in the field of biomarkers for diabetes combined with coronary heart disease

Active Publication Date: 2021-03-19
青岛山大齐鲁医院(山东大学齐鲁医院(青岛))
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the relationship between lncRNA and diabetes with coronary heart disease. Therefore, the study of lncRNA differential expression in the blood of patients with diabetes with coronary heart disease will provide a new direction for the diagnosis and treatment of diabetes with coronary heart disease

Method used

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  • Biomarker for diabetes mellitus complicated with coronary heart disease
  • Biomarker for diabetes mellitus complicated with coronary heart disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] extract serum RNA

[0022] (1) Collect blood from 30 healthy people (set as group A), blood from 22 patients with diabetes mellitus and coronary heart disease (set as group B), 25 blood from patients with pure coronary heart disease (set as group C) and 20 patients with simple diabetes blood (set to group D);

[0023] (2) Place the plasma of each group in a centrifuge, centrifuge at 3000rpm / min for 15min, and take the supernatant to obtain serum;

[0024] (3) Take 250ul of serum from each group and put them in a centrifuge tube, add 750ul of Invitrogen TRIzol reagent, blow and mix well, and let stand at room temperature for 5 minutes;

[0025] (4) Add 200ul of chloroform, shake and mix well, and let stand at room temperature for 15min;

[0026] (5) Place the centrifuge tube in a low-temperature centrifuge and centrifuge at 12,000 rpm / min at 4°C for 15 minutes;

[0027] (6) Draw the supernatant into a new centrifuge tube, add an equal volume of isopropanol, mix well, ...

Embodiment 2

[0033] Preparation of cDNA of serum in each group

[0034] (1) Removal of genomic DNA

[0035] The reaction system is as follows:

[0036] Reagent Usage amount 5×gDNA Eraser Buffer 2.0 μl gDNA Eraser 1μl Total RNA 1μg RNase Free dH2O up to 10 μl

[0037] The reaction conditions were as follows: 42°C for 2 minutes, 4°C.

[0038] (2) Reverse transcription reaction

[0039] The reaction system is as follows:

[0040] Reagent Usage amount Reaction solution of step (1) 10μl PrimeScript RT Enzyme Mix I 1.0μl RT Primer Mix 1.0 μl 5×PrimeScript Buffer 2 4.0 μl RNase Free dH2O 4.0 μl Total 20 μl

[0041] The reaction conditions are as follows: 37°C for 15 min; 85°C for 5 s; 4°C.

Embodiment 3

[0043] Detection of differential expression of AC021188.4 in each group

[0044] (1) Design specific primers for AC021188.4, the primer sequences are as follows:

[0045] AC021188.4 Upstream primer: 5'-CACATCGAGGCTCTAGTGACC-3'

[0046] AC021188.4 downstream primer: 5'-GCTTGCCAGCACGATTTACAA-3'

[0047] GAPDH upstream primer: 5'-CCCATCACCATCTTCCAGGAG-3'

[0048] GAPDH downstream primer: 5'-TTCTCCATGGTGGTGAAGACG-3'

[0049] (2) Perform fluorescent quantitative PCR reaction:

[0050] Reagent Usage amount SYBR Green Premix Ex Taq (2×) 10μl upstream primer 0.4μl downstream primer 0.4μl cDNA template 2μl sterile water 7.2μl Total 20μl

[0051] Reaction conditions: 95°C for 30s; 95°C for 5s, 60°C for 35s, 40 cycles; 70°C for 45s.

[0052] (3) Use 2 -△△Ct Methods Calculate the differential expression level of AC021188.4 in each group.

[0053] Experimental results such as figure 1 As shown, it can be seen from the result...

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Abstract

The invention provides a biomarker for diabetes mellitus complicated with coronary heart disease, and belongs to the technical field of biomedicine. Through RNA extraction, it is found that compared with serum of a healthy human body, the expression quantity of the gene AC021188.4 in serum of a patient suffering from the diabetes mellitus complicated with the coronary heart disease is remarkably reduced, while the expression quantity of the gene AC021188.4 does not remarkably change in serum of a patient suffering from the simple coronary heart disease and serum of a patient suffering from thesimple diabetes mellitus, and therefore the AC021188.4 can serve as a gene marker for the mellitus complicated with coronary heart disease.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a biomarker of diabetes mellitus complicated with coronary heart disease. Background technique [0002] With the gradual aging of the population, the incidence of coronary heart disease and diabetes is increasing year by year. Relevant research data show that the incidence of coronary heart disease in diabetic patients is 2-4 times that of normal people. Compared with patients with pure coronary heart disease, coronary artery lesions in diabetic patients with coronary heart disease are more diffuse and severe. It can be seen that diabetes combined with coronary heart disease has become a major disease that seriously threatens human life and health. Therefore, realizing the early diagnosis of diabetes mellitus with coronary heart disease will help realize the early treatment of diabetes mellitus with coronary heart disease and improve the cure rate of patients. [0003] lnc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11C12N15/113
CPCC12Q1/6883C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 王兆平王革王颖翠
Owner 青岛山大齐鲁医院(山东大学齐鲁医院(青岛))
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