RNA extraction and RT-qPCR Analysis:
Real-time quantitative polymerase chain reaction (RT-qPCR) was performed
to assess the expression of the ARG1 gene and NOS1 gene in serum using
specific primers. The extraction of total RNA was carried out using the
Steady Pure Universal RNA Extraction Kit obtained from Accurate
Biotechnology, Hunan Co., Ltd. Reverse transcription was performed using
the Evo M-MLVRT Kitt kit from the same manufacturer. Subsequently,
qRT-PCR was conducted on the CFX96 Real-Time System by Bio-Rad,
utilizing the SYBR Green Supermix provided by Accurate Biotechnology,
Hunan Co., Ltd. The 2-ΔΔCt method was
employed to calculate the relative gene expression levels, with the
housekeeping gene GAPDH serving as the internal control. The designed
primers are listed in Table S1.