Method for rapidly evaluating composition effect of estrogen and antiestrogen
An anti-estrogen and estrogen technology, applied in biochemical equipment and methods, material excitation analysis, microbial determination/inspection, etc. Observation and operation, the effect of many samples and large number of eggs laid
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Embodiment 1
[0027] Example 1 , Using zebrafish embryos to evaluate the compound effect of 17β-estradiol and tamoxifen
[0028] Test drug: 17β-estradiol (E 2 ), tamoxifen, all purchased from sigma company in the United States.
[0029] Preparation method: The standard product was dissolved in 0.1% acetone to prepare 1g / L mother liquor and stored at 4°C. The laboratory was diluted with ultrapure water to the required concentration.
[0030] Animal: Zebrafish.
[0031] Reagents: RT-PCR kit, purchased from Bao Biological Engineering (Dalian) Co., Ltd.
[0032] The specific steps of the test method are as follows:
[0033] 1. Obtaining zebrafish embryos: match 10 pairs of male and female fish at a ratio of 1:1 or 1:2, put them in a special zebrafish hatching box, separate the male and female fish with a partition, and place the broodstock under light after 12 hours , Start spawning within half an hour, suck out the eggs with a straw at the end of spawning, check for fertilization under a microscope, di...
Embodiment 2
[0047] Example 2 , Application of zebrafish embryos to evaluate the compound effect of o,p'-DDT and tamoxifen
[0048] Test drugs: o,p'-DDT, tamoxifen, all purchased from sigma company in the United States.
[0049] Preparation method: The standard product was dissolved in 0.1% acetone to prepare 1g / L mother liquor and stored at 4°C. The laboratory was diluted with ultrapure water to the required concentration.
[0050] Animal: Zebrafish.
[0051] Reagents: RT-PCR kit, purchased from Bao Biological Engineering (Dalian) Co., Ltd.
[0052] Test method: Compared with the test method of Example 1, the difference is that the selected mixture ratio of estrogen and anti-estrogens is: o,p'-DDT is 100μg / L, tamoxifen is 10μg / L, 50μg respectively / L and 250μg / L.
[0053] Test results: At the morphological level of embryonic development, o,p'-DDT and tamoxifen did not show any neutralizing or balancing effects, but a certain degree of toxic additive effects, such as Figure 5 , Image 6 with Fig...
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