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A 20 l mix, containing 8 l of cDNA sample, 10 l of SYBR Green PCR Master Mix (Thermo Fisher Scientic), and 2 l of 10 mM forward primer and reverse primer mixture was used to performed Quantitative RT-PCR with a StepOne Plus Real-time PCR system (Applied Biosystems, Foster City, CA, United States)

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The long-term use of such treatments may have significant side effects, which should also be investigated and considered