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Reactions were prepared in a 96-well plate at 10 µL total volume with 1X One-Step Master Mix, 200-400 nM forward and reverse primer, 20-35 ng total RNA diluted in sterile water, and 1X qScript One-Step Reverse Transcriptase

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The involvement of these distant organs in IR injury significantly complicates clinical outcomes, increasing the risk of multi-organ failure and underscoring the need for targeted therapeutic interventions