To evaluate the clinical efficacy of flexible ureteroscope (F-URS) combined with holmium laser lithotripter in treating renal calculi in horseshoe kidney.From November 2010 to December 2013, the medical history and charts of sixteen patients (mean age 42.9 ± 11.6 years, range 26-66 years), including 13 males and 3 females were analyzed retrospectively. Mean stone burden was 29 ± 8 mm (range 17-42 mm2). Mean stone digitized surface area (DSA) was 321 ± 94 mm2 (range 180-538 mm2). Under spinal anesthesia in a modified lithotomy position with the head down, rigid ureteroscope was placed firstly into the ureter to reach the level of the pelvis, a zebra guide wire was inserted and following the removal of the rigid ureteroscope, an ureteral access sheath was positioned along the guide wire, then passed the URF P-5 flexible ureteroscope into the renal cavities over the guidewire. After locating the stones, holmium laser lithotripsy was performed.The average operative time was 92 ± 16 minutes (range 74-127 min.). No major complications were encountered. Ten patients obtained stone-free status with one session, four obtained stone-free status after two sessions. Single session stone-free rate was 62.5%, overall stone-free rate was 87.5%. Two patients have small residual stones in the lower pole.F-URS combined with holmium laser lithotripter and nitinol basket, is safe and effective in dealing with moderate stone diameter (<30 mm) in HSKs with high clearance rates and low complication rates.