The transports of the dynamic biochemical signals in the non-reversing pulsatile flows in the mixing microchannel of a Y-shaped microfluidic device are analyzed. The results show that the mixing micro-channel acts as a low-pass filter, and the biochemical signals are nonlinearly modulated by the pulsatile flows, which depend on the biochemical signal frequency, the flow signal frequency, and the biochemical signal transporting distance. It is concluded that, the transfer characteristics of the dynamic biochemical signals, which are transported in the time-varying flows, should be carefully considered for better loading biochemical signals on the cells cultured on the bottom of the microfluidic channel. The transports of the dynamic biochemical signals in the non-reversing pulsatile flows in the mixing microchannel of a Y-shaped microfluidic device are analyzed. The results show that the mixing micro-channel acts as a low-pass filter, and the biochemical signals are nonlinearly modulated by the pulsatile flows, which depend on the biochemical signal frequency, the flow signal frequency, and the biochemical signal transporting distance. It is said that, the transfer characteristics of the dynamic biochemical signals, which are transported in the time-varying flows , should be carefully considered for better loading biochemical signals on the cells cultured on the bottom of the microfluidic channel.