In-vivo screening method for examining thyroid disruption and application
A thyroid and screening technology, applied in the field of environmental toxicology, can solve problems such as low sensitivity, large error, and inaccurate test results, and achieve high sensitivity, small error, and accurate test results
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Embodiment 1
[0051] Raising and breeding of embodiment 1 experimental animals
[0052] Feeding conditions
[0053] The water quality conditions are as follows: tap water filtered by activated carbon has a total residual chlorine of 0, an iodine concentration of 2-4 μg / L, a pH value of 6.5-7.0, a dissolved oxygen concentration of >5 mg / L, and a water hardness (calculated as CaCO3) of about 150 mg / L.
[0054] Lighting conditions: full-wavelength fluorescent lamps create natural lighting conditions, and the light and dark photoperiod is 12h / 12h.
[0055] Temperature conditions: The temperature of the water is kept at 22±1°C.
[0056] Xenopus adult frog bait: chopped pork liver: amphibian special feed (Hong Kong, China) at a ratio of 1:1, 3 times a week.
[0057] Xenopus tadpole bait: live artemia, fed 3 times a day.
[0058] induced breeding
[0059] Xenopus laevis is self-cultivated in this unit, and the earliest frog was purchased from Nasco, USA. Adult female and male frogs of Xenopus...
Embodiment 2
[0060] Example 2 Methimazole interferes with the thyroid gland of Xenopus laevis at stage NF48
[0061] Dilute the stock solution of methimazole into dechlorinated tap water to prepare test solutions with serial concentrations of 1mg / L, 5mg / L and 10mg / L, and set a blank control at the same time. Select Xenopus laevis tadpoles at the NF48 stage and randomly distribute them to each treatment tank, with 20 tadpoles per tank, 18L exposure solution, and two parallels for each treatment. Exposure was done in a semi-static manner with water changes 2-3 times a week. NF48 stage tadpoles were exposed for 4 weeks, and the developmental period composition was counted at one week of exposure. When the exposure was terminated, the tadpoles were anesthetized in MS-222, and the length of the tadpole’s hind limbs, the length from the mouth to the cloaca, developmental stage, and body weight were measured; In the fixative solution, the thyroid histological structure analysis is to be done.
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Embodiment 3
[0069] Example 3 Sodium perchlorate interferes with the thyroid gland of Xenopus laevis at NF48 stage
[0070] NF48 stage tadpoles were exposed to sodium perchlorate (32μg / L, 63μg / L, 125μg / L) for one week, and the developmental stages were distributed. The chi-square statistics showed that after NF48 stage tadpoles were exposed for one week, all doses significantly inhibited the development of tadpoles. Developmental stage distribution (p image 3 shown.
[0071] At the end of exposure, after NF48 stage tadpoles were exposed to sodium perchlorate (32μg / L, 63μg / L, 125μg / L) for 4 weeks, the distribution of developmental stages of tadpoles using chi-square statistics showed that 32μg / L, 63μg / L, 63μg / L, 125μg / L sodium perchlorate significantly inhibited the metamorphosis of tadpoles (p Figure 4 shown.
[0072] At the end of exposure, after NF48 stage tadpoles were exposed for 4 weeks, the higher doses of 63 μg / L and 125 μg / L significantly inhibited the growth of tadpole hindlimbs,...
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