In this paper, hemicelluloses from bamboo (Phyllostachys pubescens Mazel) stem aged six months were sequentially extracted with hot water, 2% KOH, and 5% NaOH. The water-soluble hemicelluloses H 1 , and four alkali-soluble hemicellulosic fractions H 2 , H 3 , H 4 and H 5 were obtained, achieving a total yield of 59.60% based on the original hemicelluloses. Sugar composition analysis showed that the hemicelluloses were mainly composed of xylose (44.39–72.71%), arabinose (26.36–51.87%), ribose (0.93–2.72%), and uronic acid (0.29–5.27%). The structures of the hemicelluloses were determined to be mainly arabinoxylan, using the FT-IR and NMR techniques. The AFM images of fraction H 3 revealed a distribution of spherical nanoparticles with different sizes, while the fraction H 2 had helix rod and random coil feature. The SEM characterizations indicated that the fraction H 1 displayed spherical particles while the fraction H 3 had mainly flat particles at high magnification. Thermal stability was also analyzed using TG–DTG method and first-order kinetics model.