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The Lop Nur region, in the east part of Tarim Basin, was an important transportation junction between west and east, north and south Eurasia. However, previous studies on prehistoric human activity have concentrated mostly on the Bronze Age, whereas that during the Stone Age remains largely unresearched. Here, we present a new direct evidence of human activity in the late Pleistocene, recorded on a grinding stone buried in a lacustrine sediment section of the Lop Nur region. The grain size distribution of the sediment section indicates that the site was probably in the center of a lake with weak hydrodynamic environment. Therefore, the stone artifact can only be carried to here by people instead of river and it was never move as soon as left here. Results of radiocarbon dating, the evident stratigraphic relations between the stone artifact and the sedimentary formation, indicate that the human activity could extend to approximately 13 ka BP. Furthermore, the results of starch-grain and use-wear analyses suggest that ancient humans gathered seeds of Triticeae, roots, and tubers and used the grinding stone to simply process selected plant as plant foodstuffs during this period. It implies that the environmental conditions in the river delta of the Lop Nur were inhabitable during the late Pleistocene.