Application of bubble walnut shell extract in the preparation of antitumor drugs
A technology of soaking walnuts and extracts, which is applied in the field of medicine to achieve the effects of simple extraction process, low cost, stable ingredients and drug efficacy
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Embodiment 1
[0018] Example 1 : Cytotoxicity test of walnut shell extract on human lung cancer cells (NCI-H292, NCI-H661)
[0019] 1.1 Preparation of soaked walnut shell extract:
[0020] Pickled walnuts (Juglans sigillata) were collected from Yangbi County, Yunnan Province, and were identified as mature fruits of Juglandaceae (Juglandaceae) by Professor Xia Conglong from the Department of Pharmacognosy, Dali College. The shells of the fruits were dried in the sun for later use.
[0021] 1.1.1 Preparation of bubble walnut shell extract PHTK-DEF
[0022] Crush 1 kg of dried soaked walnut shells, add 800 ml of sodium hydroxide solution (1 mol / liter), soak overnight at room temperature, and filter. Add 700 milliliters of sodium hydroxide solution (1 mol / liter) again, stand for 2 hours, filter, and combine the filtrates. Add glacial acetic acid to the filtrate to pH=5, let stand for 1 hour, and filter. Add 1 times the amount of ethanol (95%) to the filtrate, let it stand for 1 hour, and fi...
Embodiment 2
[0043] Example 2 : Cytotoxicity test of walnut shell extract on human liver cancer cells (HepG-2, Huh-7)
[0044] 2.1 Preparation of soaked walnut shell extract:
[0045] The same method as in Example 1.1 was used to prepare PHTK-DEF and PHTK-D, extracts of foamed walnut shells.
[0046] 2.2 Cytotoxicity test:
[0047] 2.2.1 Instruments and materials
[0048] (1) Instrument: with embodiment 1.
[0049] (2) material: with embodiment 1
[0050] (3) Cell lines: human liver cancer cells HepG-2 and Huh-7, purchased from the Cell Bank of Kunming Institute of Zoology, Chinese Academy of Sciences.
[0051] 2.2.2 Method
[0052] (1) Cell inoculation: HepG-2 and Huh-7 cells were respectively cultured in DMEM medium containing 10% inactivated FBS in a 5% carbon dioxide incubator at 37°C. After trypsinization, centrifuge, discard the supernatant, and adjust the cell concentration to 1×10 with complete medium 4 per milliliter, inoculate in a 96-well plate, 150 microliters per well...
Embodiment 3
[0063] Example 3 : Effect of bubble walnut shell extract on normal liver cells (L-02)
[0064] 3.1 Purpose of the experiment:
[0065] The toxicity of the extract was evaluated through the growth effect of the walnut shell extract on normal hepatocytes (L-02 cell line).
[0066] 3.2 Preparation of soaked walnut shell extract:
[0067] The same method as in Example 1.1 was used to prepare PHTK-DEF and PHTK-D, extracts of foamed walnut shells.
[0068] 3.3 Cytotoxicity test:
[0069] 3.3.1 Instruments and materials
[0070] (1) Instrument: with embodiment 1.
[0071] (2) Material: with embodiment 1.
[0072] (3) Cell line: normal human hepatocyte L-02, purchased from the Cell Bank of Kunming Institute of Zoology, Chinese Academy of Sciences.
[0073] 3.3.2 Method
[0074] (1) Cell inoculation: L-02 cells were cultured in DMEM medium containing 10% inactivated FBS in a 37°C, 5% carbon dioxide incubator, and the above-mentioned cells in the logarithmic growth phase were d...
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