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The aim of this study was to explore the primary chemical properties and anti-fatigue effect in vivo of Pholiota nameko polysaccharide (PNP). Through UV-visible spectrum, the absorption peaks of proteins, nucleic acids and pigments were not found. The organic functional groups of polysaccharides (3,289.97, 1,584.72, and 1,045.23 cm-1 so on) were measured by IR spectroscopy. The PNP was a semi-crystalline or non-crystalline substance, possessed a three-dimensional lump structure with a smooth, dense surface and amorphous structure accordi