In vivo screening method and application for detection of thyroid disturbance

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

Active Publication Date: 2019-07-23
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The first object of the present invention is to provide an in vivo screening method for detecting thyroid interference, and the second object of the present invention is to provide the application of the above-mentioned in vivo screening method for detecting thyroid interference in detecting thyroid interference, so as to relieve the existing The technical problems of low sensitivity, large error and inaccurate detection results exist in the technology

Method used

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  • In vivo screening method and application for detection of thyroid disturbance
  • In vivo screening method and application for detection of thyroid disturbance
  • In vivo screening method and application for detection of thyroid disturbance

Examples

Experimental program
Comparison scheme
Effect test

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.

...

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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Abstract

The invention provides an in-vivo screening method for examining thyroid disruption and application, which relate to the technical field of environmental toxicology. The in-vivo screening method for examining thyroid disruption provided by the invention exposes xenopus laevis tadpoles at the stage of NF48 to chemical contaminant to examine the metamorphic development index of the xenopus laevis tadpoles, the in-vivo screening method for examining thyroid disruption is highly efficient and highly sensitive, the error is small, and the examination result is accurate.

Description

technical field [0001] The invention relates to the technical field of environmental toxicology, in particular to an in vivo screening method and application for detecting thyroid interference. Background technique [0002] Although methods have been developed to assess the effects of chemicals on the development of the rodent cerebellum in laboratory studies, the difficulty of manipulating embryos in utero hinders the broad availability of the method. In teleosts, due to the differences in the thyroid system itself and the regulatory mechanism, it is difficult to unify the research on thyroid interference in various fish species, and it is difficult to analogize it to humans due to the differences between the thyroid system and mammals. [0003] In amphibians, except that the hypothalamus secretes CRH (corticotropin releasing hormone) instead of TRH (thyrotropin-releasing hormone) in mammals, the composition of the thyroid system is similar to that in mammals. very consist...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/50
CPCG01N33/5008
Inventor 李培峰张银凤常文光王曼
Owner QINGDAO UNIV
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