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1 Remove Melanotan II vial from refrigeration and allow to reach room temperature (1520 minutes) 2 Clean vial top with alcohol swab and allow to air dry completely 3 Using a sterile syringe, draw the calculated volume of bacteriostatic water (BAC water) 4 Inject BAC water slowly down the side of the vial do not aim directly at the powder 5 Gently swirl in circular motions DO NOT shake vigorously as this degrades the peptide 6 Allow 23 minutes for full dissolution solution should be clear and colorless 7 Draw calculated dose into an insulin syringe for subcutaneous injection 8 Inject into abdomen, thigh, or upper arm

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All operations in this study were approved by the Medical Technology Clinical Application and Research Ethics Committee of Hangzhou First People's Hospital (approval number: KY-20211105-0066-01), and each participant's written informed consent was obtained

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siRNA gene knockdown validation was determined 72 hours after siRNA transfections via qPCR