Application of ophicalcitum extract and method for preparing same
A technique of extracting stamens, used in the field of chemistry and medicine
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Embodiment 1
[0015] Embodiment 1: The preparation steps of stamen water extract and extract with anticancer activity:
[0016] Sampling: According to the requirements in "Rock and Mineral Analysis", use the interactive method and the quartering method to divide the stamens to ensure that the samples are uniform. Take out the stamens by quartering, wash and air-dry the stamens.
[0017] Pulverization: After the purified stamens are pulverized by a high-speed universal pulverizer, the powder is passed through a 100-200 mesh sieve and transferred to an oven, dried at a constant temperature of 105°C for 2 hours, and then placed in a drying oven to cool down for later use.
[0018] Extraction: Accurately weigh 100g of stamen powder dried at constant temperature, put it in a 2000mL round bottom flask, add 1000ml of deionized water at 1:10, reflux extraction 2-4 times, each time for 1-2h, filter while hot, and filter the filtrate Concentrate, set the volume in a 100ml volumetric flask, add water...
Embodiment 2
[0021] Embodiment 2 stamen (produced in Henan), serpentine fiber (produced in Sichuan) water decoction extract to human liver cancer cell HepG2, human liver cancer cell SMMC-7721, cervical cancer cell Hela, colon cancer cell SW-480 in vitro Cytostatic studies.
[0022] In vitro cancer cell inhibition experiments:
[0023] A: Prepare cancer cell suspension, take each tumor cell in logarithmic growth phase, digest with 0.25% trypsin, and prepare 5×10 cells with culture medium 4 cells / ml of cell suspension, seeded in 96-well culture plate, 100 μL per well.
[0024] B: Cells were cultured at 37°C, 5% CO2 and saturated humidity for 24 hours.
[0025] C: Filter the drug solution to be tested through a 0.45 μm filter membrane, add the drug according to the concentration of different crude drug amounts, 100 μL per hole, set 5 duplicate holes, use the normal cancer cell group as the negative control group, add the control group, etc. Measure fresh culture medium. Cells were culture...
Embodiment 3
[0038] Example 3 Study on the in vivo anticancer activity of stamen fiber nanoparticles on mouse liver tumor H22.
[0039] A: Culture and passage of H22 tumor cells
[0040]Before H22 cells were inoculated, they were mixed with DMEM+10% calf serum at 37°C and 5% CO 2 Subcultured to a good state, made 2×10 7 cell suspension per ml, inoculated in the peritoneal cavity of healthy Kunming mice, and passaged in ascites.
[0041] B: Establishment of H22 liver cancer mouse solid tumor model
[0042] Under aseptic conditions, extract the ascites of H22 mice inoculated for 7-9 days with a syringe, dilute with sterile saline to adjust the cell volume to 1×10 7 per ml, inoculated into the armpits of 50 mice, each with 0.2 ml, observed the growth of the mice, and made a solid tumor model.
[0043] C: Animal grouping and dosing
[0044] 24 hours after inoculation, they were randomly divided into 5 groups, 10 rats in each group, and administered by intragastric administration at regula...
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