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Quantitative real-time PCR reaction was performed with QuantStudio 3 Real-Time PCR System (Thermo Fischer Scientific) using the following cycling parameters: 60C for 10 min, 95C for 10 min, 40 cycles of 95C for 15 s, and 60C for 1 min, and a dissociation stage of 95C for 15 s, 60C for 1 min, 95C for 15 s, and 60C for 15 s