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Mindfulness-based interventions have emerged as unique approaches for addressing a range of clinical and subclinical difficulties such as stress, chronic pain, anxiety, or recurrent depression. Moreover, there is strong evidence about the positive effects of yoga practice on stress management and prevention of burnout among healthcare workers. The aim of this study was to conduct a single-arm clinical trial to assess the effectiveness of an intervention based on mindfulness-based stress reduction and yoga in improving healthcare workers’ quality of life. Healthcare workers of two hospitals in Rome were enrolled in a 4-week yoga and mindfulness course. Four questionnaires were administered at different times (Short Form-12 (SF-12), State-Trait Anxiety Inventory (STAI) Y1 and Y2, and Positive and Negative Affect Schedule (PANAS)) to evaluate the efficacy of the intervention. Forty participants took part to the study (83.3 %). The Mental Composite Score-12, that is part of the quality of life assessment, passed from a median of 43.5 preintervention to 48.1 postintervention (p = 0.041), and the negative affect passed from a score of 16 in the preintervention to 10 in the postintervention (p < 0.001). Both the forms of the STAI questionnaires showed a decrease after the intervention. Yoga and mindfulness administered together seem to be effective to reduce stress and anxiety in healthcare workers, providing them with more consciousness and ability to manage work stressful demands.