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These results suggested that the model was successfully established ( FIGURE 1 To determine the effect of hypothyroidism on the estrus cycle, we detected the vaginal smears for each phase including proestrus (P), estrus (E), metestrus (M), and diestrus (D) (Supplementary Figure S1A), the results showed that the estrus cycle of rats in the hypo group became irregular, primarily manifested as a significantly prolonged estrus cycle, extended from 4.37 0.18 days to 6.48 0.21 days, while the proportion of proestrus in the overall estrus cycle was decreased from 9.37% 1.88%5.43% 1.69%, and the proportion of estrus tended to increase (Table 1