Peripheral blood and middle ear effusion (MEE) from PATIENTS with OME (PC) and healthy volunteers (NC)
Repeatedly OME patients (PC Group) and obviously out of ear disease and systemic abnormal immune response to disease (after IgE and clinical symptom screening) of healthy people (NC Group) were recruited to collect 2 ml venous blood. Signed informed consent was obtained from patients and healthy people and this study was approved by the ethnic committee of Shenzhen Longgang District Hospital of Otolaryngology (Shenzhen Institute of Otolaryngology).
Peripheral blood mononuclear cells (PBMC) from the remaining blood were isolated by density gradient centrifugation and RNA was extracted from the harvested cells using TRIzol according to the manufacturer’s instructions. CDNA was synthesized by Sangon Biotech (Shanghai, China) and amplified by QRT-PCR (Applied Biosystems, Foster City, California, US). The data were analyzed using relative gene expression methods and the GAPDH Ct values in the samples were standardized. Each sample shall be measured in triplicate.
Primer sequences used for PCR amplification are as follows: