Journal of Medical Internet Research - International Scientific Journal for Medical Research, Information and Communication on the Internet #Preprint #PeerReviewMe: Warning: This is a unreviewed preprint. Readers are warned that the document has not been peer-reviewed by expert/patient reviewers or an academic editor, may contain misleading claims, and is likely to undergo changes before final publication, if accepted, or may have been rejected/withdrawn. Readers with interest and expertise are encouraged to sign up as peer-reviewer, if the paper is within an open peer-review period. Please cite this preprint only for review purposes or for grant applications and CVs (if you are the author). Background: Smartphones have integrated into our society and are expected to serve as tools to improve health outcomes. In the summer of 2016, Pokémon Go, a location-based augmented reality game for smartphones, was released and attracted attention from the perspective of health, especially with its potential to increase physical activity (PA). A few studies have compared objectively-measured step counts before and after the release of the game; however, they were conducted over a short study period and evaluated only young people. Objective: Forty-eight players and 192 non-players aged ≥38 years were matched for sex, age group, and PA level; they were respondents to a questionnaire randomly sent to citizens who were given free pedometers by Yokohama city. Their play status was identified through the questionnaire. To investigate the change in step counts before and after the release of Pokémon Go according to play status, two-way repeated measures ANOVA was performed. Step counts one month before the release of the game were compared with those eight months after the release. In addition, subgroup analysis according to sex, age group, PA level, and subjective health status was conducted. Methods: Among citizens who was given a free pedometer by Yokohama City, 48 players and 192 non-players, matched for sex, age group and PA level, were selected. They were aged 38 or older and their play statuses were identified through a questionnaire randomly sent to those citizens. To investigate the change in step counts before and after the release of Pokémon GO by play status, two way repeated measures ANOVA was used. Step counts before the release of 1 month were compared with those after the release of 8 months. In addition to the analysis of all samples, subgroup analysis by sex, age group, PA level and subjective health status were conducted. Results: The mean ages of players and non-players were 56.6 (±9.7) years and 57.3 (±9.6) years, respectively, and the mean of baseline step counts of players and non-players were 7776.5 (±2773.3) and 7947.1 (±2849.2), respectively. There was no significant difference in the age and baseline step counts according to a t-test. In the analysis of all samples, the interaction between play status and time effect was significant for three of eight months after release. In the subgroup analyses, the interaction was significant for five months in men, eight months in the 55–64-year-old group, three months in workers, two months in the active group in PA level, and four months in participants with subjectively good health. The interaction was significant for only one month at most in other subgroups. Conclusions: The present study confirmed a difference in step counts between players and non-players before and after the release of Pokémon GO. According to our analysis, step counts were higher until six months after the release. There was no decrease in the step counts of players even in cold months, despite the decrease in step counts of non-players. In subgroup analyses, players were more likely to be men, aged <55 years, workers, active, and subjectively in good health.