Objective: To observe the medium-and long-term clinical efficacy and safety of radioactive 125I seed implantation for refractory malignant tumors based on CT-guided 3D template assisted technique. Methods: Twenty-five patients with refractory malignant tumors who received radioactive 125I seed implantation based on CT-guided 3D template assisted technique in our hospital from August 2016 to March 2017 were selected for retrospective analysis. The postoperative adverse reactions were recorded, and the number of puncture needle and particles used in the operation and dosimetric parameters, as well as postoperative physical strength scores and tumor response were statistically analyzed, the overall survival time and survival rate were counted, and the effect and prognosis were explored. Results: All patients were successfully completed 125I seed implantation, and no serious complications occurred. The average number of implanted puncture needles was 17 (19.12±13.00), and the median number of particles was 52 (55.12±32.97). D90 in the postoperative CTV (93.24±15.70) Gy was slightly lower than preoperative CTV (93.92±17.60) Gy (P>0.05). D90 in the postoperative PTV (142.16±22.25) Gy was lower than preoperative PTV (145.32±23.48) Gy (P>0.05). The tumor responses at 6 months postoperatively were: CR: 20% (5/25), PR: 48% (12/25), SD: 24% (6/25), PD: 8% (2/25), CR+PR: 68%, local control rate: 92% (23/25). The 6-, 12-, and 24-month survival rates were 100%, 88%, and 52%, respectively. The KPS physical strength score was gradually recovered, and increased to the highest value at 12 months after seed implantation, and then slightly decreased and the average score was still above 90 points. There were 1 case of pneumothorax during operation, and 2 cases with superficial malignant tumors developed skin ulceration. Multivariate analysis of prognosis showed that tumor sites and types were independent risk factors affecting survival, and the number of needles and particles, and template type were not factors that affected prognosis. Conclusions: 3D template combined with CT-guided radioactive 125I seed implantation improves the rational distribution of radiation dose in tumor target area because of the achievement of accurate radioactive 125I particle implantation, and it has less complications, and can further extend the overall survival and quality of life in patients with refractory malignant tumors.