Fig 1: rTMS treatment improved motor functions and increased the proportion of peripheral Tregs in PD patients. A Study flow chart. Sixty patients with PD were screened for eligibility in this clinical study between Sep 2021 and Jan 2022. A total of 6 PD patients were ultimately excluded. Finally, the 54 PD patients were randomized (1: 1) to divided into sham and rTMS group. Demographic data, basic examinations including electroencephalography, medication dosage before and after rTMS or sham treatment were carried out in both groups of patients. Tregs, aTreg, nTreg, proportion and the levels of IFN-γ, TNF-α, IL-17α, IL4, IL10 and TGF-β1 in the peripheral blood were measured, H&Y and MDS-UPDRS III scores were performed at the same time point before and after rTMS or sham treatment. B Tregs, and aTreg/nTreg proportion in the peripheral blood were assessed by flow cytometry prior to treatment and 1 day after the treatment. Hoehn and Yahr and MDS-UPDRS III were performed prior to treatment and on Days 13, 19, and 40 after the treatment. C No significant difference between the Mean H & Y ± SD on Days -1, 13, 19 and 40 in the sham and rTMS groups. (sham vs. rTMS groups on days -1: n = 27, P = 0.761, t = -0.306, d. f. = 52; sham vs. rTMS groups on days 13: n = 27, P = 0.761, t = -0.306, d. f. = 52; sham vs. rTMS groups on days 19: n = 27, P = 0.761, t = -0.306, d. f. = 52; sham vs. rTMS groups on days 40: n = 27, P = 0.761, t = -0.306, d. f. = 52). D There was a significant difference in the difference between the MDS-UPDRS III scores of the rTMS group and the sham group on day 13, 19, and 40 and their respective MDS-UPDRSIII scores on day -1.(rTMS vs. sham groups between days 13 and -1: n = 27, P < 0.0001, t = 15.64, d. f. = 208; rTMS vs. sham groups on 19 and -1: n = 27, P < 0.0001, t = 15.38, d. f. = 208; rTMS vs. sham groups on on 40 and -1: n = 27, P < 0.0001, t = 15.77, d. f. = 208). E The baseline of the proportion of CD4 + CD25 + CD127- Treg cells in the peripheral blood displays no significant difference between sham and rTMS group. (sham vs. rTMS groups: n = 27, P = 0.796, t = 0.26, d. f. = 52). F The peripheral blood CD4 + CD25 + CD127- Treg cells before and after sham rTMS treatment in the sham treatment group shows no significant difference. (before vs. after: n = 27, P = 0.185, t = 1.363, d. f. = 26). G The peripheral blood CD4 + CD25 + CD127-Treg cells before and after rTMS treatment in the rTMS treatment group shows significant difference. (before vs. after: n = 27, P < 0.000, t = -14.88, d. f. = 26). H Mean change in the proportion of Tregs/CD4 + T cells in the peripheral blood ± SD between days -1 and 13 in the sham and rTMS groups. (n = 27, P < 0.0001, t = 14.98, d. f. = 52). I No correlation between the change in the proportion of Tregs/CD4 + T cells and the change in MDS-UPDRSIII scores (between days -1 and 13) in sham group (n = 27, r2 = 0. 0007, p = 0.895). J Negative correlation between the change in the proportion of Tregs/CD4 + T cells. and the change in MDS-UPDRSIII scores (between days -1 and 13) in rTMS group. (n = 27, r2 = 0.8261, p < 0.0001). rTMS = repetitive transcranial magnetic stimulation; H & Y = Hoehn-Yahr; MDS UPDRS III = Movement Disorder Society Unified Parkinson's Disease Rating Scale part III; SD = Standard Deviation; Data are presented as mean ± SD; two-tailed unpaired t test (C, D, E, H); paired sample t test (F, G); linear regression analysis (I, J). ****P < 0.0001. Each data point represents an individual subject. Comparisons with no asterisk had a P > 0.05 and were considered not significant