This paper presents an integrated Re-Os isotope and trace element investigation of the Neoproterozoic to Cambrian organic-bearing black mudstones in the eastern Three Gorges, the Yangtze Craton. Their sedimentary environmental evolution and geological significance are then discussed. It shows that 1) initial Os isotopic ratios of fine-grained sedimentary rocks from the strata display regular variations along the stratigraphic column; 2) the Nantuo tillite displays higher initial Os isotopic ratio than that of the overlying cap carbonate, while the overlying successive strata show an increase trend with several fluctuations; 3) geochemical features of U/Th ratio and δU value for the samples suggest that the Liantuo sandstone and Nantuo tillite were formed within oxidation environments; and 4) negative correlation between Y/Ho ratio and δCe value for the black mudstones is likely indicative of variable involvement of seafloor hydrothermal activity during some sediments deposited.