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Objectives. To assess the feasibility and utility of a newly developed wireless portable ultrasound “UProbe- 2” for puncture guidance in percutaneous nephrolithotomy (PCNL). Materials and Methods. The UProbe-2 consists of a probe with integrated ultrasound circuit boards inside, an intelligent terminal that installed specific ultrasound software, and a puncture trestle. The probe can connect to an intelligent terminal via WI-FI. It was used to guide puncture in PCNL in 31 patients who suffered kidney stones or large impacted proximal upper ureteral stones with hydronephrosis from June 2016 to July 2017. Patients’ characteristics as well as parameters during operation were collected. Results. PCNL was successfully performed in all patients with the guidance of the UProbe-2. The mean (standard deviation) puncture time was 3.6 (0.9) minutes, and the median operative duration was 50 (35-80) minutes. The primary stone clearance rate was 87% (27/31). No major intraoperative or postoperative complications occurred. Conclusion. The UProbe-2 ultrasound is safe and effective for puncture guidance during PCNL according to this initial study. It could provide a new and alternative choice for guidance of PCNL. However, more clinical trials especially randomized controlled trials should be performed.