Aims This study aimed to develop a Practice Programme for Upright Positions in the Second Stage of Labour to provide a reference for midwifery professionals in the standardized implementation of upright positions in clinical practice. Background The adoption of upright positions in the second stage of labour is recommended by many international organizations, but upright positions have not been widely used and their implementation varies greatly across studies. Methods The Practice Programme for Upright Positions in the Second Stage of Labour was developed under the guidance of the Medical Research Council framework for developing and evaluating complex interventions and the World Health Organization handbook for guideline development. Four stages were conducted: (1) establishing the intervention development group, (2) identifying a theoretical basis and forming a content framework, (3) evidence retrieval and synthesis and (4) refining and modelling the practice programme. Results The content framework of the Practice Programme for Upright Positions in the Second Stage of Labour was formed based on the literature review, semi-structured interviews and expert consultation, including indications and contraindications, implementation methods, observations, potential risks and precautions. According to each item, we conducted a series of systematic reviews, and summarized the available best evidence from clinical guidelines, systematic reviews and original studies. Eventually, the Practice Programme for Upright Positions in the Second Stage of Labour was developed, integrating the findings of the iterative evidence reviews and revised by stakeholders. Conclusions This study first reported the development process of the Practice Programme for Upright Positions in the Second Stage of Labour, characterized by evidence-based, iteratively processed and highly rigorous. The implications may guide researchers to embed the intervention normatively into clinical practice for improving maternal and infant outcomes. Implications for Nursing Management The Practice Programme for Upright Positions in the Second Stage of Labour could facilitatesystematic management of labour positions and guide midwives in the successful implementation of upright positions.