Background and Purpose: Tui Na is an alternative and non-invasive intervention, based on TCM theory of the meridian and acupoint, and has been widely used in clinical treatment. Tui Na has been proved effective and safe for poststroke upper limb dysfunction. Yet the specific neural mechanism associated with Tui Na is still limited. This study was aimed to explore the changes in spontaneous brain activity, as a reflection of cortical plasticity, induced by Tui Na in post-stroke patients, with the application of rs-fNIRS assessment. Materials and Methods: A total of 18 participants, including 10 stroke patients and 8 healthy controls (HC), underwent a rs-fNIRS assessment before and after a session of 30-minute Tui Na on Yangming Meridian. The optrodes were placed upon the bilateral frontal and parietal cortex. With the preprocessed fNIRS data, the fractional amplitude of low-frequency fluctuation alterations between the two groups were calculated, standardized and analyzed. Results: At baseline assessment, no significant difference in zfALFF values was observed between post-stroke patients and HCs. After Tui Na, compared to HCs, stroke patients showed higher spontaneous activity in SMA, SAC, and ipsilesional SM1, and lower in the contralesional PMC. Furthermore, Changes of zfALFF values in contralesional SM1 after Tui Na was negatively correlated with FMA-UE score in stroke patients. Conclusion: With resting-state fNIRS, this study investigates for the first time the spontaneous neural changes in post-stroke patients following Tui Na on Yangming Meridian. The preliminary results indicate that Tui Na may enhance plasticity of the somatosensory cortex associated with clinical upper limb function.