Supplementary Material for: Urinary Metabolomics Study on Acupuncture plus Wet Cupping Therapy for Post Stroke Depression: A pilot study

Objective: This study aimed to investigate urinary metabolomics alterations associated with acupuncture plus wet cupping (AC) therapy in patients with post-stroke depression (PSD). Methods: A total of 40 subjects were randomly allocated to either the acupuncture plus wet cupping group (AC group, n=20) or the acupuncture group (Acu group, n=20). Morning urine samples were collected at baseline and after 4 weeks of treatment, yielding a total of 80 samples. An additional 20 healthy volunteers served as a blank control group, from whom urine samples were also obtained. Urinary metabolites were analyzed using gas chromatography-mass spectrometry (GC-MS). Differential metabolites and related metabolic pathways were identified. Clinical efficacy was assessed using the Hamilton Depression Rating Scale-24 items (HAMD-24) at baseline and post-treatment. Correlation analysis was performed to assess the relationship between metabolic changes and clinical improvement. Results: After 4 weeks of treatment, the HAMD-24 scores in the AC group (median: 10.00, IQR: 9.00-13.00) were significantly lower than those in the Acu group (median: 18.00, IQR: 15.25-19.00), with a statistically significant between-group difference (P