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This study aimed to describe endoscopic anatomy of the seminal tract and summarize our experience of transutricular seminal vesiculoscopy (TSV) guided by real-time transrectal ultrasonography (TRUS) in managing persistent hematospermia.A total of 281 consecutive patients with persistent hematospermia who underwent TSV with or without real-time TRUS were enrolled in this single-center,prospective,observational study.The median follow-up period was 36.5 (range: 8.0-97.5) months.TSV was successfully performed in 272 (96.8%) patients.The approach of a 4.5/6 F rigid vesiculoscope entering the seminal tract was categorized into four types on the basis of endoscopic presentation of the ejaculatory duct orifice and verumontanum.Seven (2.6%),74 (27.2%),64 (23.5%),and 127 (46.7%) patients had Types Ⅰ (through the ejaculatory duct in the urethra),Ⅱ (through the ejaculatory duct in the prostatic utricle),Ⅲ (transutricular fenestration through a thin membrane),and Ⅳ (real-time transrectal ultrasound-guided transutricular fenestration) approach,respectively.In patients who successfully underwent surgery,bleeding occurred in the seminal vesicle in 249 (91.5%) patients.Seminal vesiculitis,calculus in the prostatic utricle,calculus in the ejaculatory duct,calculus in the seminal vesicle,prostatic utricle cysts,and seminal vesicle cysts were observed in 213 (78.3%),96 (35.3%),22 (8.1%),81 (29.8%),25 (9.2%),and 11 (4.0%) patients,respectively.Hematospermia was alleviated or disappeared in 244 (89.7%) patients 12 months after surgery.Fifteen patients had recurrent hematospermia,and the median time to recurrence was 7.5 (range: 2.0-18.5) months.TSV guided by TRUS may contribute to successful postoperative outcomes in managing persistent hematospermia.