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Anxiety is one of the problems in patients undergoing invasive procedures. One way to control anxiety is through psychological preparation. This study was aimed to evaluate the effect of psychological preparation on anxiety level in patients undergoing upper gastrointestinal endoscopy. Methods The present study consisted of 98 patients which were referred to Ahvaz Golestan Hospital for upper gastrointestinal endoscopy. They were randomly (table of random numbers) divided into a control group and an experimental group. All of the participants filled out Spielberg State–Trait Anxiety Inventory (STAI). The experimental group was psychologically prepared for endoscopy (given information about endoscopy and behavioural intervention), and then both groups were asked to complete STAI again an hour before endoscopy. The collected data were analysed through Mann–Whitney, Kruskal-Wallis, and Chi-square tests using SPSS 17.0. Results Before the intervention was carried out, there was no significant difference between the two groups in terms of the rate of state and trait anxiety (p > 0.05). After the intervention, the rate of state and trait anxiety decreased significantly (p < 0.05) in experimental group. Conclusion In patients subjected to endoscopy, psychological preparation was effective in reducing their anxiety and thus this can be considered as an efficient method in decreasing anxiety.