Aqueous extracts of Chinese herbs and composition and application thereof
A water-based, biological technology, applied in the direction of microorganisms, animal cells, tissue culture, etc., can solve the problem that patients cannot be treated smoothly
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preparation example 1
[0062] Preparation Example 1: Primary culture of human dermal fibroblasts
[0063] Skin samples were obtained with the subject's consent. Skin samples were cut into skin fragments 0.5 to 1 mm cubic in size. Subsequently, the skin fragments were placed in DMEM containing FBS, collagenase and dispase, and incubated at 37° C. for 16 to 18 hours. Centrifugation was performed at 1,500 rpm for 5 minutes. Then, the supernatant was removed to replace the broken cells and skin pieces supplemented with FBS, and the broken skin pieces and cells were evenly distributed in Erlenmeyer flasks, and cultured at 37°C. Cells were subcultured when the cell growth density reached 80%. The cells of the third to sixth passages were used for the experiments described below.
preparation example 2
[0064] Preparation Example 2: Preparation of Chinese herbal medicine extract
[0065] Astragalus and Alisma were purchased from traditional Chinese medicine shops. Earth dragons were purchased from area fishing supply stores. Bitter melon was purchased from the regional fruit and vegetable market.
[0066] 100 grams of Astragalus polyphylla was added to 500 grams of secondary deionized water, followed by boiling at normal pressure for 2 hours. Weigh to get 210 grams of Astragalus polysequence suspension.
[0067] 100 grams of Alisma was added to 500 grams of secondary deionized water, followed by boiling under normal pressure for 2 hours. Weigh 242 grams of Alisma suspension.
[0068] 100 grams of balsam pear fruit was added to 500 grams of secondary deionized water, followed by boiling at normal pressure for 2 hours. Weigh 150 grams of bitter gourd suspension.
[0069] 100 grams of unprocessed whole worms of Earthworm were added to 500 grams of secondary deionized water...
Embodiment 1
[0072] Example 1: Unilateral treatment of cells with extracts
[0073] Take quantitative fibroblasts obtained in Preparation Example 1, plant them in a 96-well flat culture dish, and in the presence of the individual Chinese herbal medicine extracts prepared in Preparation Example 2, culture them in a medium containing 0.5% heat-deactivated calf serum (FBS; Gibco, NY, USA) in Duchenne's Modified Eagle's Medium (DMEM; Sigma Chemical Co., St. Louis, MO, USA) for 3 to 6 days. The control group was the fibroblasts of Preparation Example 1 cultured in DMEM containing 0.5% or 1.0% FBS. The supernatant and cells were harvested and analyzed separately.
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