Application of serum extracellular vesicle miR-503-5p in preparation of diagnostic kit for type-2 diabetes
A technology of type 2 diabetes and diagnostic kits, applied in the determination/testing of microorganisms, biochemical equipment and methods, etc.
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Embodiment 1
[0031] Example 1 Dynamic changes in the degree of methylation of the miR-503-5p promoter region during natural aging.
[0032] Experimental method: Use the bioinformatics software Methprimer to retrieve the promoter region of 2500bp upstream of miR-503-5p, and analyze the possible methylation modification sites; extract DNA from embryonic mice and aged mice, as well as human plasma DNA, The degree of methylation at specific sites was identified using MSP technology.
[0033] Experimental results: if figure 1 As shown, there are a large number of methylation modifications in the promoter region of miR-503-5p in mouse and human DNA ( figure 1A), Methylation-specific PCR (methylation-specific PCR, MSP) shows that the methylation site at -2397bp ~ -2213bp upstream of miR-503-5p has specific methylation products after birth in mice, This indicates that the methylation level of this site has a tendency to gradually increase during development ( figure 1 B), similarly, our MSP res...
Embodiment 2
[0034] Example 2 The relationship between serum extracellular vesicle miR-503-5p content and the pathogenesis of type 2 diabetes.
[0035] Experimental animals: Male C57BL / 6J mice were purchased from the Institute of Model Animals, Nanjing University. All animal experiments were approved by the Experimental Animal Management Committee of Nanjing Medical University, and all experimental animals were kept in a barrier facility in the Experimental Animal Center of Nanjing Medical University in a clean environment with a temperature of (21±2)°C and a humidity of (35±2)°C. )%, 12h: 12h day and night without lighting, free to eat and drink, the drinking water is the sterile water prepared by the experimental animal center.
[0036] Experimental method: We established a high-fat model when the mice were 4 weeks old, and continuously monitored the changes in their body weight and random blood sugar, and detected the content of miR-503-5p in serum extracellular vesicles before and afte...
Embodiment 3
[0045] Example 3 The relationship between human serum extracellular vesicle miR-503-5p content and type 2 diabetes.
[0046] Experimental materials: We collected blood samples from people undergoing physical examination in provincial government hospitals in Jiangsu. The inclusion criteria for the healthy adult group were people aged 18-40 with no history of diabetes and other diseases, and the inclusion criteria for the elderly group were those over 60 years old without diabetes. People with a history of diabetes, the inclusion criteria for the elderly diabetic group are people over 60 years old with diabetes but without diabetic complications. The specific information of the three groups of people is shown in Table 1-3.
[0047] Table 1 Group information of healthy adults
[0048] Numbering gender age BMI (kg / m 2 )
Fasting blood glucose (mg / dL) history of diabetes 1 male 37 20.6 96.84 none 2 Female 29 22.5 96.12 none 3 Fema...
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