For the safety issues of night long-distance bus drivers, a test system that real-time continuously acquires vehicles driving parameter, traffic environmental images as well as ECG physiological state of drivers under night-time driving conditions is designed. The impact of continuous driving time, night-time conditions and others on the driver's heart electrical parameters and driving behaviors is studied. The experiments were carried out on 11 drivers and the total journeys were over 11,800 km. The experiments' results indicate that: The long-distance bus drivers. The driver's average heart rate shows a downward trend as the driving time extends. And the average heart rate while driving is 13~18 times higher than the average heart rate while resting. Under night highway free-flow conditions and the highest safety speed 100 km/h of the passenger transport company regulations, the driver's 50%, 75% and 95% percentile speed are 93.6 km/h, 99.3 km/h, 103.2km/h respectively, and 50%, 75% and 95% of the throttle pedal position are 75%, 85%, 90% of the largest pedal trip respectively.