Objective:To study the T/N ratio difference and trend of99mTc-HL91 hypoxia imaging before and after brain malignant glioma with radiotherapy. Methods: 85 postoperative patients with malignant glioma were detected99mTC- HL91 SPECT hypoxia imaging before and after radiotherapy in which radioactive count(ROI)in the early phase(half an hour)and delayed phase(5 hours)in the tumor area and normal tissue was tested. Results:T/N at delayed phase was statistical significance before and after radiotherapy in99mTC- HL91 SPECT hypoxia imaging(P0.05). T/N after radiotherapy at delayed phase was lower than before. All patients had no comfortable and serious toxic effect during the test. Conclusion:T/N at delayed phase was statistical significance before and after radiotherapy in99mTC- HL91 SPECT hypoxia imaging, which indicates that the radiotherapy can kill cancer cell and provides some scientific data for effect of radiotherapy on postoperative patients with malignant glioma. T/N after radiotherapy at delayed phase was lower than before, which shows that status of hypoxia declines after radiotherapy.