The dilatation curves of continuous cooling transformation at different cooling rates were determined for U75V rail steel by THERMECMASTOR-Z thermal simulator, and continuous cooling transformation curve was obtained. The influence of cooling rate on microstructure and hardness was studied. The softening behavior after isothermal deformation in the austenite region 850-1000°C but before the second pass was also studied by double-pass compression tests. The results show that the product of austenite decomposition was pearlite when the cooling rate was lower than 10°C. Troostite and martensite were gained at the cooling rate of 10°C•s-1. Only martensite was obtained when the cooling rate was in the range of 10-50°C•s-1. The hardness of the steel increased with the increase of cooling rate. Under the condition of 30% deformation and 3s-1 deformation rate, the relaxation time for completing recrystallization was shorter than 100s when the deformation temperature was higher than 1000°C. When the deformation temperature was lower than 880°C, full recrystallization was difficult to achieve even if the relaxation time was extended.