Experimental method: Select three groups of SD rats.

The first group recruited 20 SD rats and divided them into two groups: drinking ordinary water and H2 PEM water bottle every day. These rats were not given thioacetamide. This experiment is used to determine the safety of long-term use of hydrogen-rich water and whether hydrogen-rich water can improve the quality of life (Supplementary Figure S1). Concentration of H2 PEM water bottle produced by Liry will up to be 5000ppb with working 10mins,USB charging, simple and convenient, charge anytime, anywhere.

In the second group, 38 adult male SD rats (300-370 g) were selected and given thioacetamide with or without hydrogen-rich water for 5 months. The rats were divided into thioacetamide group (n = 19) and thioacetamide + hydrogen group (n = 19). All rats were given 300 mg/L thioacetamide in their drinking water daily until sacrificed. In addition, the test group took 2ml of 1.8 mg/L H2 cup daily and 2ml of regular water in the control group. After 8 weeks of treatment, 3 animals in each group were sacrificed every month, and livers were taken to test the tumorigenic effect of thioacetamide and the preventive effect of hydrogen. The animals are weighed weekly.

The third group recruited 30 adult male SD rats. Rats in each group were given the same thioacetamide water for 3 months and then stopped drinking. They were divided into normal water group and hydrogen-rich water group. This experiment is used to test whether hydrogen-rich water can block the progress of CCA.

Result

Long-term use of Hydrogen Water Maker affects the safety and quality of life. There were no examples of thioacetamide or drug-related mortality in all experiments. The first experiment focused on the safety of the use of hydrogen-rich water. Compared with the control group, the use of hydrogen water for 9 months did not cause any pathological changes in internal organs including the heart, liver and kidneys (Figure 1A). There was no significant difference in AST and ALT levels between the two groups (Figure 1B). However, during this period, we observed two exciting things: the hair of rats that drank hydrogen-rich water was whiter and brighter than that of rats that did not drink hydrogen-rich water (Figure 1C); Sperm motility is higher than the latter (Figure 1D). In addition, HE and IHC staining showed that the former rats had denser hair follicles and higher proliferation activity than the latter rats (Figure 1C).