Objective To investigate the clinical effects and safety differences of endoscopic hormone injection by tympanic membrane puncture and eustachian tube in the treatment of patients with chronic secretory otitis media. Methods From January 2015 to September 2016, 90 patients with chronic secretory otitis media in Zhoushan Hospital were chosen.They were randomly divided into two groups according to the digital table.The tympanic membrane puncture group (65 patients) was treated with hormone injection by tympanic membrane puncture and the eustachian tube group(65 patients) was treated with endoscopic hormone injection by eustachian tubeThe total clinical effective rate, the bone conduction threshold value in 1kHz, 2kHz, 4kHz and 8kHz, the ETDQ-7 score and the levels of laboratory index before and after treatment, the secondary tympanic effusion rate and the recurrence rate with follow-up of both groups were compared. Results The total clinical effective rate of the tympanic membrane puncture group and eustachian tube group were 77.78% and 93.33%, respectively.The total clinical effective rate of the eustachian tube group were significant higher than that of the tympanic membrane puncture group(χ2=9.84, P<0.05). The bone conduction threshold value in 1kHz, 2kHz, 4kHz and 8kHz of the tympanic membrane puncture group after treatment were (10.42±1.60)kHz, (12.86±2.50)kHz, (16.09±2.81)kHz, (15.26±2.68)kHz, respectively.The bone conduction threshold value in 1kHz, 2kHz, 4kHz and 8kHz of the eustachian tube group after treatment were (9.75±1.36)kHz, (11.13±2.02)kHz, (14.82±2.44)kHz, (13.78±2.11)kHz, respectively.The bone conduction threshold value in 1kHz, 2kHz, 4kHz and 8kHz of the eustachian tube group after treatment were significant lower than those of the tympanic membrane puncture group and those before treatment(t=3.10, 3.56, 3.29, 3.96, 2.87, 3.15, 2.91, 3.28, 2.67, 2.81, 2.78, 2.50, all P<0.05). The ETDQ-7 scores of the eustachian tube group after treatment were significant lower than those of the tympanic membrane puncture group and those before treatment(t=3.31, 3.87, 2.89, all P<0.05). The levels of laboratory index of the eustachian tube group after treatment were significant higher than those of the tympanic membrane puncture group and those before treatment(t=3.36, 3.77, 3.71, 4.02, 2.83, 3.06, all P<0.05). The secondary tympanic effusion rate and the recurrence rate with follow-up of the eustachian tube group were significant lower than those of the tympanic membrane puncture group(χ2=9.87, 12.33, 10.67, all P<0.05). Conclusion Compared with endoscopic hormone injection by tympanic membrane puncture, endoscopic hormone injection by eustachian tube in the treatment of patients with chronic secretory otitis media possess the advantages including relieve the clinical symptoms, improve the bone conduction threshold and eustachian tube function, regulate the laboratory indexes and reduce the risk of recurrence and effusion. Key words: Otitis media with effusion; Tympanic membrane; Punctures; Eustachian tube; Dexamethasone