Thermal analysis of biological tissues subject to a train of ultrashort pulse irradiations was made of developing a combined time-dependent radiation and conduction bio-heat transfer model. A model skin tissue stratified as three layers with different optical, thermal and physiological properties was considered. Temperature response of the skin tissue exposed to a single ultrashort pulse irradiation was firstly analyzed by the finite volume method in combination with the transient discrete ordinates method. This temperature rise was found to reach pseudo steady state within an extremely short time period in which thermal diffusion is negligible. Since the tissue properties were assumed to be constant during a train of pulse irradiation, this temperature rise subject to a single pulse can be employed for repeated pulses, In the same time, Pennes' equation was employed to study the bio-heat transfer in the meso-time scale. The effects of pulse strengths and repetition rate on the temperature response in the multi-layer skin tissue were investigated.