OBJECTIVE: This study investigated the associations between changes in autonomic nervous system (ANS) function, psychological status during the mechanical ventilation (MV) weaning process, and weaning outcomes. METHODS: In this prospective study, we recruited 67 patients receiving MV for >24h at a medical center in northern Taiwan. Patients\' ANS function, represented by heart rate variability (HRV), the rapid shallow breathing index (RSBI), anxiety, fear, and dyspnea, was repeatedly measured 10min before and 30min after undergoing a weaning trial. Forty-nine patients capable of sustaining a 2-h weaning trial were successfully weaned. RESULTS: Compared with the failed group, the success group showed significantly smaller decreases in high-frequency HRV (HRV-HF) and smaller increases in RSBI (per 10 breaths/min/L), fear, dyspnea, and anxiety in response to the weaning trial (odds ratio [OR]=2.19, 0.81, 0.69, 0.66, and 0.77, respectively; p<0.05). Multivariate analyses revealed that low-frequency HRV before weaning (OR=2.32; 95% confidence interval [CI]=1.13-4.78, p=0.02), changes in HRV-HF (OR=3.33; 95% CI=1.18-9.44, p=0.02), and psychological fear during the weaning process (OR=0.50; 95% CI=0.27-0.92, p=0.03) were three independent factors associated with 2-h T-piece weaning success. CONCLUSIONS: ANS responses and psychological distress during weaning were associated with T-piece weaning outcomes and may reflect the need for future studies to utilize these factors to guide weaning processes and examine their impact on outcomes.