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Relative mRNA expression was determined using the GoTaq 1-step real-time quantitative PCR (RT-qPCR) system (TAKARA) and qPCR using SYBR premix Ex Taq kit (TAKARA), and RT-qPCR was performed on an ABI Prism 7500 Fast Real-Time PCR System (Applied Biosystems, CA, USA)

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A significant gray zone exists between 200 and 300 pg/mL where patients may experience symptoms despite levels that appear technically normal