Application of nitazoxanide in pharmacy
A technology for nitazoxanide and medicine is applied in the application field of nitazoxanide in pharmaceuticals, which can solve the problems of poor prognosis of patients with bone metastases, and achieve the effect of low acquisition cost.
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Embodiment 1
[0048] Example 1 In vivo imaging system to observe the effect of nitazoxanide on bone metastasis of prostate cancer cells
[0049] 1. Construction of highly metastatic PC3-KQ cells in prostate cancer
[0050] 1. Conversion
[0051] Take the competent Escherichia coli out of the refrigerator at -80°C and thaw on ice; add the ligation product (K369Q) constructed in the laboratory into the competent Escherichia coli and incubate on ice; place on ice after heat shock at 42°C for 90s 2min; Add 200 μL of LB medium and 1 μL of antibiotics to the competent state in the ultra-clean bench, pipette evenly, and then coat the plate containing antibiotics; after coating the plate, culture in a constant temperature incubator at 37°C overnight.
[0052] 2. Plasmid amplification
[0053] Use a 10 μL pipette tip to pick up the monoclonal colonies on the plate and put them into EP tubes. Shake the bacteria in a shaker at 200 rpm for 12 hours, and extract the plasmids according to the plasmid e...
Embodiment 2
[0081]Example 2 microCT detection of the protective effect of nitazoxanide on bone tissue in mice
[0082] After the experiment in Example 1 was over, the mice were killed by dislocation of the cervical vertebrae, and the bone tissue was dissected and fixed with paraformaldehyde for 48 hours. The bone tissue was then immersed in a centrifuge tube containing PBS, and then small animal microCT was performed to observe the transfer of PC3-KQ cells. and bone tissue destruction, and detection of cortical bone destruction, and analysis of trabecular density (BV / TV), trabecular number (Tb.N), trabecular thickness (Tb.Th) and trabecular separation (Tb.Sp) and other indicators.
[0083] The result is as Figure 6 , Figure 7 and Figure 8 As shown, among them, Figure 6 In order to analyze the protective effect of NTZ on the bone tissue of living mice by micro-CT in the embodiment of the present invention, at the end of administration, micro-CT was performed on the PC3-KQ control g...
Embodiment 3
[0085] Embodiment 3HE staining detects the therapeutic effect of nitazoxanide on mouse bone tissue
[0086] 1. HE staining
[0087] (1) After the mice in different treatment groups were killed by cervical dislocation, the femur and tibia were separated, and the excess connective tissue and muscle tissue around them were cut off, fixed in 4% paraformaldehyde for 48 hours, and then decalcified with EDTA decalcification solution for 14-21 hours. sky;
[0088] (2) After the bone tissue is decalcified, it is dehydrated, and the procedure is as follows:
[0089] 70% ethanol 1h
[0090] 80% ethanol 1h
[0091] 90% ethanol 1h
[0092] 95% ethanol 1h
[0093] Absolute ethanol Ⅰ 1h
[0094] Absolute ethanol Ⅱ 1h
[0095] Xylene Ⅰ 1h
[0096] Xylene II 1h
[0097] Paraffin Ⅰ 1h
[0098] Paraffin II 1h
[0099] Paraffin III 1.5h
[0100] (3) Embedding: Turn on the embedding machine 2 hours in advance, preheat and melt the paraffin; soak the embedding cassette in liquid paraffi...
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