The effects on semen and the function of accessory gonad of workers after short and long term exposure to benzene, toluene, and xylene were examined. The semen and blood of 24 married workers exposed to benzene, toluene, and xylene were collected. Routine sperm characteristic, acrosin activity, and Lactate dehydrogenase C4 (LDH-C4) relative activity were detected. The results showed that benzene, toluene, and xylene were found in the blood and semen of some ex-workers at workplaces where the air concentration of benzene, toluene, and xylene exceeded the maximum allowable concentration (MAC). No such solvents were detected in the blood and semen of workers of the control group. The sperm vitality and sperm motility decreased in the exposed workers. The mean acrosin activity, γ-GT activity and LDH-C4 relative activity in the exposed workers were lower, and fructose concentration was higher than those in the control. There were negative correlations between sperm vitality, sperm activity, acrosin activity, or LDH-C4 relative activity and working history. These results suggest that the mixture of these solvents could affect the sperm and the function of accessory gonad. This might be one reason of the abnormal pregnancy outcome among the wives of workers exposed to benzene, toluene, and xylene.