To investigate the etiology, clinic characteristics, and pathogenesis of cholesterol granuloma(CG) of the middle ear, as well as to discuss the effect of surgeries. MethodsForty-three patients (43 ears) with cholesterol granuloma of the middle ear were enrolled in this retrospective study, and 42 received middle ear mastoid surgery and were confirmed by a pathologic examination post-operatively. The main clinical manifestations of CG were hearing loss (43/43), tinnitus (26/43), sense of fullness (29/43), and bloody otorrhea (12/43), with blue drum as a typical physical sign (36/43). Temporal bone high resolution computed tomography (HRCT) scan showed soft tissue shadows in 43 cases with erosion of the ossicular chain in 17 cases, which was confirmed by surgeries in 13 cases. Results 31 patients underwent a combined modality treatment of mastoidectomy, tympanotomy, and grommet insertion and other 11 patients were subjected to a combined strategy of mastoidectomy, tympanotomy, and reconstruction of the ossicular chain. The eardrums of patients became normal 1 to 2 months after the surgeries. The hearing threshold was improved (the improvement of ABgap was 25±3.5dB) and no symptoms recurred in all patients after a follow-up from half to 3 years. ConclusionThe main clinical features of cholesterol granuloma of the middle ear are hearing loss, tinnitus, sense of fullness, and bloody otorrhea, as well as blue drum. HRCT is useful for diagnosis and operation. As for the treatment of CG, mastoid surgery should be performed on, in which the affected tissues in the middle ear should be completely removed; a ventilation system of the mastoid tympani should be established.