The demanding and unpredictable nature of nursing duties, particularly night shifts, poses significant challenges to nurses' overall well-being. Night shifts, characterized by altered circadian rhythms and increased workplace demands, present unique challenges affecting both the physical and mental health of nurses. This cross-sectional study aimed to explore the psychological and physical health of nurses due to night shift duty and identify research gaps. Conducted in Dhaka Medical College Hospital and Shaheed Suhrawardy Medical College Hospital, the study included 326 purposively selected nurses from January to December 2023. Data were collected through face-to-face interviews using the Depression, Anxiety, and Stress Scale (DASS-21) and the Physical Health Questionnaire (PHQ) and were analyzed using SPSS. The majority (75.5%) of respondents were female, with a mean age of 32 years (±6.50 SD). Findings revealed that 14.4% of nurses experienced severe depression, while 8.9% suffered from extremely severe depression. Additionally, 19.9% reported severe anxiety, and 22.7% had extremely severe anxiety. Regarding stress levels, 16.6% experienced severe stress, and 6.1% faced extremely severe stress. In terms of physical health, 27.3% of nurses often struggled with sleep difficulties, 24.8% frequently suffered from headaches, 16.3% often had an upset stomach, and 23.9% experienced flu symptoms lasting more than seven days’ post-night shift duty. Females exhibited significantly higher levels of anxiety than males (χ2 = 14.813, p<0.05). A strong positive correlation was found between depression and stress (r = 0.707, p<0.001). The findings suggest that night shift duties may impair both the psychological and physical health of nurses. Therefore, hospital authorities and policymakers must be sensitized to these issues, and appropriate measures should be implemented to make night shifts more comfortable, ultimately improving nurses' clinical performance, personal health, and overall quality of patient care.