Epidemiological investigations indicate that nursing professionals experience a higher prevalence of musculoskeletal disorders (MSDs) than most other occupational groups. Most nursing activities require standing and walking for prolonged periods. Such job requirements may contribute to MSD problems in the lower extremities. Thus, wearing comfortable nursing shoes is essential to reduce the lower-extremity discomfort for clinical nurses. The objectives of this study are: (1) to evaluate three brands of commercially available nursing footwear and identify the important shoe features for adequate shoe support during nursing activities, and (2) to assess the effect of wearing compression hosiery by measuring the biomechanical, physiological, and psychophysical responses of test participants. The method of this study involved two phases. First, field observations were conducted to collect job demand data, including walking speed, and the ratios of walking, standing, and sitting. Second, an experiment was conducted to evaluate the functions of professional nursing footwear and examine the influence of compression hosiery on lower extremity discomfort relief. Measurements included electromyography (EMG) of the lower leg, joint range of motion (ROM) in the lower extremity, foot pressure, ground reaction force (GRF), and subjective discomfort ratings for the lower body and feet. Summarizing the findings of this study, comfortable footwear for nursing professionals should emphasize a footbed with arch support outside with 1.5 cm thickness of EVA materials in the metatarsal zone and heel height between 1.8 and 3.6 cm; this can minimize foot pressure distribution, impact force, and increase shin and ankle comfort. In addition, wearing compression hosiery is recommended to alleviate lower body and foot discomfort for clinical nurses.