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Objective: To evaluate the changes of pelvic floor tissue structure and function between pregnant and non-pregnant women from the view of transperineal pelvic floor ultrasound., Methods: Thirty-eight cases of women with a second singleton pregnancy and thirty-two cases of women with a first singleton pregnancy underwent transperineal pelvic floor ultrasound, and their results were compared with forty-two cases of healthy non-pregnant women., Results: The differences of bladder neck descent (BND), rectal ampulla distance and levator hiatus area (LHA) among the three groups were statistically significant (P<0.05), and the differences of BND, rectal ampulla distance, LHA between the women with a second singleton pregnancy group and non-pragnent group were statistically significant (P<0.05). The BND, retrovesical angle at rest (RVA-R) and retrovesical angle underwent Valsalva maneuver (RVA-V) in the group of stress urinary incontinence (SUI) during pregnancy were larger than those in non-SUI group, with significant difference (P<0.05), especially BND and RVA-V (P = 0.00)., Conclusion: Transperineal pelvic floor ultrasound has a high resolution of pelvic floor structure and function changes during pregnancy, and can dynamically evaluate pelvic floor function, providing a theoretical basis for early diagnosis and prevention of female pelvic floor dysfunction (FPFD) in subsequent pregnancies., Competing Interests: The authors report no conflicts of interest in this work., (© 2022 Xu et al.)