Hochu-ekki-to is a Chinese herbal medicine that has been used for unexplained male infertility with some favorable clinical results. However, the detailed mechanisms of the effect of this medicine on male infertility have not been disclosed. This study investigated the effects of Hochu-ekki-to on sperm motility of human semen. Sixteen semen samples were obtained from 14 normal volunteers. Semen samples were incubated with 1, 10, and 100 ug/mL of Hochu-ekki-to solution in 5% CO₂ 37°C for 1-6 h, and the motility was compared with that of the control groups (the untreated semen and the semen incubated with phosphate-buffered saline). The motility of sperm in 10 and 100 μg/ mL of Hochu-ekki-to solution for 1-6 h was significantly higher than that of the control groups. One microgram/milliliter of Hochu-ekki-to solution also maintained higher sperm motility after 4-6 h incubation compared with the control groups. Previous reports showed that sperm motility correlated significantly with fluorescence intensity peaks from both spermatozoa and semen plasma. However, the fluorescence intensity at 622 nm excited at 488 nm from spermatozoa fractions in each group was not significantly different. It would appear that the enhanced sperm motility induced by Hochu-ekki-to is mediated by improvement of the circumstances of semen rather than that of spermatozoa-related factors.