Objective To evaluate the clinical effect of trabeculectomy combined with amniotic membrane insertion under the scleral flap to treat patients with refractory glaucoma. Methods Thir- ty-five eyes of 28 patients with glaucoma underwent trabeculectomy combined with amniotic mem-brane insertion under the scleral flap. The filtering bleb, intraocular pressure, the success rate of the operation and complications were observed after surgery. Results After 3～24 months of follow-up, 34 eyes had type Ⅰ blebs, 9 eyes had type Ⅱ blebs, 2 eyes had type Ⅲ blebs and 3 eyes had type Ⅳ blebs. The average intraocular pressure was controlled at (13.1±2.6)mmHg after 3 months and (15.4±3.1)mmHg 12 months after surgery. Drugs were used in 2 eyes to control intraocular pressure. The total success rate of the operation was 94.3%(33/35). Five eyes with anterior chamber exudation and 1 eye diagnosed with neovascular glaucoma with a small hyphema were found post surgery, but they all recovered within a week. Conclusion Combined trabeculectomy with amniotic membrane insertion under the scleral flap can increase the success rate of glaucoma surgery.