This study aims to discover the characteristics of heart rate variability of resting and real-time motional states in healthy population. The ECG data of 16 healthy young subjects during 5-minute resting and motion periods were recorded respectively. After that, the R wave was extracted through Hilbert transformation and the RR interval time series was computed. The calculations for HRV parameters in time and frequency domains, Poincare scatter plots and the detrended fluctuation analysis were conducted. Our study finds that, the measures of real-time motion in time domain (Mean RR, SDNN and RMSSD), the measures in frequency domain (VLF, LF, HF, TP), and the measures of Poincare scatter plots are significantly less than those of resting state (P <0.001). The measure of long-term fractal exponent (alpha(2)) in real-time motion state is significantly higher than resting state (P <0.05). Between the two states, there are no statistically significant differences in other parameters of frequency domain (LF norm, HF norm, LF/HF) and short-term fractal exponent (alpha(1)), P>0.05. This study suggests that, the results can provide new characteristic signatures and statistical evidence for sport medicine or rehabilitation medicine.