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Quantitative real-time PCR (qRT-PCR) was performed using 2 sybr Green qPCR MasterMix (Sevenbio, Beijing) and the ABI StepOnePlus real-time PCR system (Applied Biosystems, United States), with -actin as the internal reference

Nitrous oxide misuse can cause vitamin B12 deficiency and absorption issues, which can lead to nerve damage, paralysis, chronic numbness, and loss of coordination

The researchers utilized advanced mass spectrometry and RNA-sequencing to map the exact metabolic dysfunctions driving these conditions, uncovering striking evidence of severe oxidative stress and mitochondrial lipid oxidative damage mediated by glutathione peroxidase 4, as documented in the PNAS metabolic study

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