Introduction: Mental health plays a key role in an individual's quality of life and general well-being, as highlighted in the World Health Organization's characterization of mental health. The main goal of this research is to compare the frequency of stress, anxiety and depression among nursing graduate students at the Faculty of Health Studies in Rijeka who listen to lectures in rotations compared to nursing graduate students at the Faculty of Medicine in Pula who listen to lectures every week. The specific goal is to examine the frequency of depression, anxiety and stress among graduate nursing students in Rijeka and Pula with regard to gender. Subjects and methods: A cross-sectional survey was conducted with 100 respondents. Respondents are of both sexes, aged from 22 to 60 years. The research was conducted with a questionnaire for the assessment of depression, anxiety and stress (Depression, Anxiety and Stress Scale), DASS-21. The questionnaire was standardized and translated into Croatian. Results: Every tenth respondent had very serious symptoms of depression, and 22% of respondents had serious symptoms. Half of the respondents had serious (14%) or very serious (36%) symptoms of anxiety, while almost a third of the respondents (32%) were under serious stress, and 6% were under very serious stress. 23% of respondents requested the help of a psychologist or other health professional, while 13% of respondents used anxiolytics. 12.0% of women and no men had very serious signs of depression. 41.2% of men and 22.9% of women have normal symptoms of anxiety. Very serious symptoms of stress are present equally in men (5.9%) and women (6.0%), but while serious symptoms of stress are shown by a third of women (33.7%). Conclusion: The results showed that there is no statistically significant difference in symptoms of anxiety and stress with regard to the way of studying. There is no significant difference in the occurrence of stress, anxiety and depression in graduate nursing students with regard to gender. Students who attend regular classes are statistically significantly less depressed than students who attend classes during the work week.