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Background: Job stress, the most important factor in reducing organizational productivity and the cause of psychosomatic disorders, today is more prevalent among the healthcare staff. Therefore, the aim of the present study was to assess the level of job stress among the nursing staff of Kashan hospitals during 2009. Materials and Methods: This descriptive cross-sectional study was performed on 190 nurses of Kashan hospitals using the Osipow questionnaire consisted of two parts: demographic data and six stress-inducing work roles (role overload, role insufficiency, role ambiguity, role boundary, responsibility and physical environment). In addition, job stress was divided into four (subnormal, normal, moderate and severe) levels. Results: Results indicated that the 95.3% of nursing staff were found to have a normal level of job stress. No significant relationship was seen between the job stress and gender; however, there was a significant relation between the role insufficiency, role boundary, responsibility and physical environment and gender. Moreover, no significant relationship was found between the job stress and some factors (e.g. age, education, marital status, income and type of hospital). The most frequent job stressors among the men and women were responsibility (90.2%) and role ambiguity (40.9%), respectively. Conclusion: Role ambiguity, responsibility and workload are the major sources of stress among the nursing staff. [ABSTRACT FROM AUTHOR]