Spasticity is a major contributor to pain, disabilities and many secondary complications after stroke. Investigating the effect of spasticity on neuromuscular function in stroke patients may facilitate the development of its clinical treatment, while the underlying mechanism of spasticity still remains unclear. The aim of this study is to explore the difference in the neuromuscular response to passive stretch between healthy subjects and stroke patients with spasticity. Five healthy subjects and three stroke patients with spastic elbow flexor were recruited to complete the passive stretch at four angular velocities (10°/s, 60°/s, 120°/s, and 180°/s) performed by an isokinetic dynamometer. Meanwhile, the 64-channel electromyography (EMG) signals from biceps brachii muscle were recorded. The root mean square (RMS) and fuzzy entropy (FuzzyEn) of EMG recordings of each channel were calculated, and the relationship between the average value of RMS and FuzzyEn over 64-channel was examined. The two groups showed similar performance from results that RMS increased and FuzzyEn decreased with the increment of stretch velocity, and the RMS was negatively correlated with FuzzyEn. The difference is that stroke patients showed higher RMS and lower FuzzyEn during quick stretch than the healthy group. Furthermore, compared with the healthy group, distinct variations of spatial distribution within the spastic muscle were found in the EMG activity of stroke patients. These results suggested that a large number of motor units were recruited synchronously in the presence of spasticity, and this recruitment pattern was non-uniform in the whole muscle. Using a combination of RMS and FuzzyEn calculated from high-density EMG (HD-EMG) recordings can provide an innovative insight into the physiological mechanism underlying spasticity, and FuzzyEn could potentially be used as a new indicator for spasticity, which would be beneficial to clinical intervention and further research on spasticity.