Objective: To investigate the influence of sustained-release fluorouracil implants on the healing process of the bilioenteric anastomosis in rabbits, so as to provide data related to the safety and feasibility of its clinical application. Methods: Forty healthy New Zealand rabbits were equally randomized into experimental group and control group. Rabbits in both groups underwent bilioenteric anastomosis, and those in experimental group received sustained-release fluorouracil implantation near the anastomotic stoma, while those in control group received the empty granules instead. In both groups, the tissues around the anastomotic site were harvested on postoperative day (POD) 3, 7, 14, 21 d respectively, and the general conditions of the abdominal cavity and anastomotic stoma were observed, van der Ham adhesion scoring for the anastomotic stoma was performed, the anastomotic bursting pressure (ABP) was measured, and the histopathological changes and collagenous connective tissues at the anastomotic stoma were examined. Results: In experimental group, 18 rabbits survived, one died of postoperative bile leakage and one died of abdominal infection; 19 rabbits survived in control group and one died of bile leakage, and there was no statistical difference in mortality between the two groups (P>0.05). The general healing statuses of the two groups were similar, and there was no significant difference in van der Ham score or ABP at each time point between the two groups (all P>0.05). The histopathological examinations (HE staining and Mallory’s trichrome staining) demonstrated that the two groups showed a similar pattern regarding the inflammatory reactions, neovascularization, and collagen fiber hyperplasia around the anastomotic stoma after operation. Conclusion: Sustained-release fluorouracil implant exerts no obvious influence on the healing process of the bilioenteric anastomosis in rabbits, so it can be used as a safe adjuvant chemotherapeutic method for prevention and treatment of postoperative local recurrence and metastasis of biliopancreatic malignant tumors.