design: The samples were transported to sequencing companies on dry ice for sequencing. Total RNA was extracted via advanced molecular biology equipment. After RNA extraction, a Nanodrop 2000 and Agilent 2100 were used to determine the purity, concentration and integrity of the RNA. After the sample was tested, a library was constructed. After the library was constructed, a Qubit 3.0 fluorescence quantification instrument was used for preliminary quantification. The concentration required was 1 ng/l or above. The Qsep400 high-throughput analysis system was subsequently used to detect the inserted fragments of the library. The effective concentration of the library (library effective concentration>2 nM) was accurately quantified by the QPCR method to ensure the quality of the library. Fastp software was used to remove reads containing adapters and low-quality reads to obtain clean data.