Establishment of zebra fish osteoporosis diagnostic standard
A standard technique for the diagnosis of osteoporosis, applied in the field of biomedicine, can solve problems such as time-consuming, no prompts, complicated staining procedures, etc., and achieve the effect of simple operation, accurate and reliable results, and fast imaging speed
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Embodiment 1
[0041] Example 1 Establishment of a diagnostic method for zebrafish osteoporosis
[0042] 1. Configuration of the solution
[0043] Prednisolone solution: Weigh 3.6 g of prednisolone, dissolve it in 10 mL of dimethyl sulfoxide (DMSO), dissolve it fully, dilute to 1 L of distilled water, and store in a sealed container with a concentration of 10 mmol / L. Dilute to 150 μmol / L before use.
[0044] 0.03% MS222 (ethyl m-aminobenzoate methanesulfonate): Weigh 30 mg of MS222, dissolve in 100 mL of distilled water, seal and store at 4°C.
[0045] 2. Establishment of zebrafish osteoporosis criteria
[0046] (1) Select 12 adult female zebrafish with normal development from 3 to 5 months and divide them into two groups. One group is placed in culture water as normal group, and the other group is placed in 150μmol / L prednisolone for induction and fed once a day. The brine shrimp were replaced with fresh induction solution every 2 days; after 21 days of feeding, they were euthanized in 0...
Embodiment 2
[0060] Example 2 Drug evaluation of zebrafish osteoporosis model
[0061] (1) Osteoporosis model group: 12 normal adult female zebrafish from 3 to 5 months were selected and divided into two groups. One group was induced with 150 μmol / L prednisolone, fed with common feed once a day, and changed every 2 days. One fresh induction solution; one group was fed a diet containing 200 μg / g FA, and the fresh induction solution was changed every two days;
[0062] After induction and feeding for 21 d, they were euthanized in 0.03% MS222, washed twice and fixed in 4% paraformaldehyde for 24-48 h.
[0063] (2) Cut out the caudal body segment of the zebrafish and store it in 70% alcohol.
[0064] (3) Micro-CT scanning: Scan the sagittal plane of the caudal body of zebrafish with SCANO MEDICALμCT 100, select the caudal body C1-C4 for three-dimensional reconstruction, obtain the bone mineral density (BMD), and record.
[0065] Scanning parameters are as follows: X-ray source is 55kV, 200μA...
Embodiment 3
[0069] Example 3 Drug evaluation of zebrafish osteoporosis model
[0070] (1) Osteoporosis model group: 12 normal adult female zebrafish from 3 to 5 months were selected and divided into two groups. One group was induced with 150 μmol / L prednisolone, fed with common feed once a day, and changed every 2 days. One fresh induction solution; one group was fed a diet containing 2 mg / g FA, and the fresh induction solution was changed every two days;
[0071] After induction and feeding for 21 d, they were euthanized in 0.03% MS222, washed twice and fixed in 4% paraformaldehyde for 24-48 h.
[0072] (2) Cut out the caudal body segment of the zebrafish and store it in 70% alcohol.
[0073] (3) Micro-CT scanning: Scan the sagittal plane of the caudal body of zebrafish with SCANO MEDICALμCT 100, select the caudal body C1-C4 for three-dimensional reconstruction, obtain the bone mineral density (BMD), and record.
[0074] Scanning parameters are as follows: X-ray source is 55kV, 200μA, ...
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