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Assessment of plasma levels Determination of mouse insulin levels using Elabscience mouse insulin enzyme-linked immunosorbent assay kit (Wuhan Elarite Biotechnology Co., Ltd.) ( Measurement of aortic stiffness by in vivo pulse wave velocity Under isoflurane anesthesia (100%, oxygen flow 1.75%), velocity waveforms were acquired at the aortic arch using a Vevo2100 high-resolution small animal ultrasound machine (VisualSonics, Canada) ( In vivo aortic stiffness was assessed by pulse wave velocity (PWV)