Complex vitamin D for preventing and treating fungal infection and application of complex vitamin D
A multivitamin and fungal infection technology, applied in the field of medicine, can solve the problems of difficulty in fungal infection, fungal drug resistance, low price, etc., and achieve the effects of low cytotoxicity, thinning of bacterial fur, and reduced spore germination rate.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0031] Embodiment 1 liquid-based diluent method drug susceptibility test (MIC 90 test)
[0032] Before the experiment, the three Aspergillus strains (A. fumigatus, Aspergillus flavus, and Aspergillus niger) were continuously replanted twice in YPD solid medium at 37°C, and Cryptococcus neoformans were cultured in YPD liquid medium at 37°C and 180rpm / min. 24h, to ensure the vigor and purity of the strains to ensure the vigor and purity of the strains. After the Aspergillus spores activated twice were eluted with sterile double distilled water containing 1% Tween 80, the Aspergillus spores and Cryptococcus neoformans were diluted with double distilled water, and then counted with a hemocytometer to determine the spore concentration. -1640 dilution to adjust the concentration of bacterial solution is 2 × 10 5 cfu / ml spare. Referring to the microdilution method of the CLSI M38-A protocol, add 100 μL of double-diluted multivitamin D to the 1st to 10th wells of the 96-well plate ...
Embodiment 2
[0036] Embodiment 2 in vitro antibacterial experiment
[0037] 1. Antibacterial experiment on Aspergillus
[0038] Prepare YPD solid medium containing different concentrations of complex vitamin D: mix vitamin D2 and vitamin D3 in different mass ratios, dissolve in DMSO to prepare a stock solution with a concentration of 50 mg / ml, and store at 4°C for later use.
[0039] Preparation of Aspergillus spore suspension: Before the experiment, the three Aspergillus strains were inoculated twice in YPD solid medium in a constant temperature incubator at 37°C to ensure the viability and purity of the strains. The twice-activated Aspergillus fumigatus spores were eluted with sterile double distilled water containing 1% Tween80, and Aspergillus spore suspensions of different concentrations were prepared with sterile double distilled water (10 5 cfu / ml, 10 4 cfu / ml, 10 3 cfu / ml) to carry out spot plate experiment, promptly take 3 μ l of spore suspension each time and spot on the prepa...
Embodiment 3
[0043] Example 3 Cytotoxicity Experiment
[0044] The cytotoxic effect of compound vitamin D (the mass ratio of vitamin D2 and 1,25-dihydroxyvitamin D3 is 1:2) on liver cancer cells (HepG2 cells) was detected by CCK-8 assay. Discard the culture medium for liver cancer cells with a growth confluence of about 90%, wash twice with PBS, add an appropriate amount of trypsin-EDTA, and store in 5% CO 2 , Incubate in a humid cell incubator at 37°C for 2 to 3 minutes. When the cells are plump and oval, add an appropriate amount of complete medium to stop the digestion, centrifuge, remove the supernatant, and collect the cells. Add an appropriate amount of regular cell complete medium, resuspend the cells, blow and mix gently with a pipette, count the cells, and use regular cell complete medium to adjust the cells to 5×10 4 / ml, and pipette evenly. The cell suspension was seeded into a 96-well plate at an amount of 100 μl / well, and 3 plates were planted (three replicates). Each 96-we...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com