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The core issues that need to be focused on in future research are optimal stimulation parameters for different IBS subtypes, combined regimens of VNS with other therapies (e.g., low FODMAP diet, prokinetics, and antidepressants), key targets to be validated in mechanistic studies of IBS-D, and so on

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Histochemical and immunohistochemical staining Tissue cryosections were fixed for 15 min, incubated with SA--gal staining at 37 C overnight in the dark according to the manufacturers instructions (Solarbio, Beijing, China)

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