Use of Liriope muscari Bailey C in pharmacy
A technology of Ophiopogon japonicus and inhibitors, which is applied in the field of pharmaceutical application of Ophiopogon japonicus C, can solve the problems that the survival period cannot be better extended and has not been reported.
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Embodiment 1
[0016] Example 1 Detection of toxicity of DT-13 to cells
[0017] (1) Material:
[0018] Instrument: Ultra-clean workbench (Suzhou Aikelin Purification Equipment Co., Ltd.), constant temperature CO 2 Incubator (Heraeus, Germany), enzyme-linked immunoassay (BIO-RAD, USA), inverted biological microscope (OLYMPUS, Japan), plate shaker (Jiangsu Guangming Experimental Instrument Factory).
[0019] Reagents: RPMI1640, F12 medium (GIBCO), trypsin (SIGMA), newborn calf serum (HYCLONE), MTT (SIGMA), DMSO (SIGMA).
[0020] Cell lines: human breast cancer cells MDA-MB-435, MDA-MB-231, MCF-7.
[0021] (2) Method:
[0022] 1. Take a bottle of cells that are in good condition in the exponential growth phase, add 0.25% trypsin digestion solution, digest to shed adherent cells, and count 2~4×10 4 Pcs / ml to make cell suspension.
[0023] 2. Inoculate the cell suspension on a 96-well plate, 180μl / well, and place constant temperature CO 2 Incubate in an incubator for 24 hours.
[0024] 3. Change the...
Embodiment 2D
[0036] Example 2 The effect of DT-13 on cell adhesion in hypoxic state
[0037] OBJECTIVE: To detect changes in the ability of human breast cancer cells to adhere to basement membrane under hypoxia in vitro
[0038] Materials: Six-well plate, 96-well plate, PBS, 1% gelatin, 2% BSA, serum-free medium, 10% calf serum medium, digestion solution (0.25% trypsin), MTT reagent, DMSO
[0039] method:
[0040] 1. Preparation of Gelatin Board:
[0041] 1. Weigh a certain amount of gelatin powder, add 1g / 100ml amount of hot PBS to dissolve it, take advantage of the hot spot into a 96-well plate, 30μl / well, lay flat and shake to make the gelatin flatten the bottom of the well, be careful not to have bubbles .
[0042] 2. Leave it overnight, suck out the gelatin, and wash it with PBS.
[0043] 3. Block it with 2% BSA for 1 hour and then suck it off.
[0044] 4. Add serum-free medium, 20-30μl / well, equilibrate for more than 1 hour, and then aspirate.
[0045] Second, cell culture
[0046] 1. Vi...
Embodiment 3
[0071] Example 3 The effect of DT-13 on cell migration ability in hypoxic state
[0072] Use Boyden Chamber Transwell to detect cell migration ability. The device is divided into internal and external rooms. The bottom of the inner chamber is a polycarbonate filter membrane (pore size 8 μm), which is first coated with 1% gelatin, and then equilibrated with serum-free culture medium for 1 hour. Human breast cancer cells (1.5×10 4 Each cell / 100μl) was suspended in a culture medium containing 2% FBS and different concentrations of the test compound and seeded into the inner chamber. Add 0.6ml of culture medium with or without VEGF (20ng / ml) to the outer chamber. After incubating at 37°C for 6 hours, discard the culture medium in the wells and fix the cells with 90% ethanol for 10 minutes. Use a cotton swab to gently wipe away the non-migrated cells in the upper layer of the inner chamber, stain with 0.1% crystal violet at room temperature for 10 minutes, rinse the excess dye with PBS...
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