Soluble insulin receptor as type 2 diabetes detection biomarker
An insulin receptor, type 2 diabetes technology, applied in the fields of medicine and biology, can solve problems such as reducing the level of INSR and exacerbating insulin resistance
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Embodiment 1
[0030] Example 1 The amount of BACE1 protein and the enzymatic cleavage activity of INSR in patients with type 2 diabetes and co-patients with type 2 diabetes and cognitive impairment are up-regulated.
[0031] 1. Collection of samples
[0032] All samples were collected by the First Affiliated Hospital of University of Science and Technology of China from January 2018 to January 2019 and were pathologically diagnosed as 56 cases of type 2 diabetes and 61 cases of cognitive impairment. Type 2 diabetes and cognitive impairment A total of 97 cases of patients and 57 cases of normal healthy elderly plasma samples. There were no differences in age and sex composition among the four groups. All plasma samples were processed in the same way, including numbering, centrifugation, supernatant plasma fraction and storage under the same conditions. This study has been approved by the Ethics Committee of the First Affiliated Hospital of University of Science and Technology of China, wit...
example 2
[0040] Example 2: Significant increase of sINSR level in plasma of patients with type 2 diabetes, type 2 diabetes and cognitive impairment; sINSR content is positively correlated with the content of BACE1 in plasma and the enzymatic cleavage activity of BACE1 to the synthetic INSR polypeptide; sINSR The content is positively correlated with the clinical indicators of type 2 diabetes.
[0041] 1. Experimental materials
[0042] Anti-INSR alpha monoclonal antibody (83-7) (Thermo Scientific, MA5-137640); biotinylated Anti-INSR beta monoclonal antibody (18-44) (Abcam, ab983); Lipo3000 (Invitrogen); pCDNA-INSR plasmid , pEGFP-BACE1 plasmid; ultrafiltration tube (Millipore, UPC905008); magnetic globin G (Thermo Fisher, 10003D); 0.45 μm PVDF (GE Healthcare Lifescience, 10600023); Corning microtiter plate (Costar, 42592); antibody coating solution (Candor-Bioscience, 120125); blocking solution (Thermo Scientific, 37515); ELISA amplification system (PerkinElmer, NEP116001EA); TMB Supe...
example 5
[0053] Example 5: The ratios of Aβ40 and Aβ42 / 40 only show differences in patients with cognitive impairment (including patients with simple cognitive impairment and patients with type 2 diabetes and cognitive impairment) but not in patients with simple type 2 diabetes, and cannot Differentiate cognitive impairment associated with type 2 diabetes from other types of cognitive impairment.
[0054] 1. Experimental materials
[0055] Fujirebio INNO-BIA plasma Aβ forms kit (#REF 81578), Luminex 200 (Luminex Corporation, Texas, USA)
[0056] 2. Specific operation
[0057] The first day: Wash the plate once with 225 microliters of washing solution, add the diluted microspheres to the wells, 100 microliters per well, and drain the liquid. Plasma was diluted in sample dilution at a ratio of 1:2 and added to the wells, 75 microliters / well, and 25 microliters of diluted secondary antibodies were added at the same time, and incubated overnight at 4 degrees. The next day: Drain the liq...
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