Objective: To investigate the feasibility and efficacy of retrograde tracing along the cystic duct to prevent bile duct injury (BDI) during laparoscopic cholecystectomy (LC).  
Methods: Three hundred and twenty-four patients undergoing LC were selected. The approach of retrograde tracing along the cystic duct was performed in all patients during operation, namely, after the supposed cystic duct was isolated, retrograde dissection closely along this duct toward the gallbladder was performed, and if this  duct was verified to be the true cystic duct, it was clamped and then cholecystectomy was performed, but if this was not the cystic duct, the anatomical pathway was changed to seek the cystic duct for prevention of BDI up to the completion of cholecystectomy.
Results: Of the 324 patients, the supposed cystic duct was verified to be the actual cystic duct or its safety site in 319 cases, was found actually to be the common hepatic (bile) duct in 3 cases, and aberrant right posterior hepatic duct opening into the cystic duct in 2 cases. LC was uneventfully performed in all the 324 patients. The average operative time was 64.3 (25–210) min, and the average blood loss was 7.3 (2–150) mL.Postoperative minor bile leakage and fluid collections in the gallbladder region occurred in one patient, which were cured by puncture drainage and wound dressing. No BDI was found in the other patients. The average length of postoperative stay was 2.6 (1–13) d. No jaundice or abdominal pain was noted in the patients during one-moth follow-up.
Conclusion: Retrograde tracing along the cystic duct for prevention of BDI during LC is safe and reliable as well as easy to perform. It can be used as a supplement to other preventive methods.