“…Based on the results of our pre-experiments and the results of water environment research and previous toxicological studies, we determined that the Cu concentration was 50 μg L −1 and the NP concentration was 1000 μg L −1 in this study. [13][14][15][16]23 The control group (control) and three treatment groups were set up, and the treatment groups included the Cu exposure group (Cu 2+ , 50 μg L −1 ), the NP exposure group (NPs, 1000 μg L −1 ) and the group with co-exposure to NPs and Cu (NPs + Cu, NPs, 1000 μg L −1 , Cu 2+ , 50 μg L −1 ). The required NPs were added to 20 mL test tubes and the pH was adjusted to 7 ± 0.1 with 0.1 mmol L −1 HCl and 0.05 mmol L −1 NaOH to obtain the stock solution for the NP group with a final concentration of 100 mg L −1 .…”