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Quantitative realtime polymerase chain reaction (qRTPCR) Total RNA was isolated using Trizol LS (Invitrogen, USA) following the manufacturers protocol

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It effectively combats oxidative stress in the testes and spermatozoa by scavenging free radicals, reducing lipid peroxidation, and bolstering the endogenous antioxidant system, thereby preserving sperm quality, motility, membrane integrity, and mitochondrial activity 17,18

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This catabolic effect is a standard biological response to rapid weight loss