Preparation of nucleic acid and traditional Chinese medicine composite aqueous solution for rapidly enhancing autoimmunity of animals
An aqueous solution and immunity technology, which is applied in the direction of drug combination, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problems that other uses of CpGODNs have not been reported, and achieve rapid enhancement of animal autoimmune response , increase the intensity of immune response, the effect of convenient application
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[0024] In the present invention, the composition includes the water extract of Chinese medicine, the water extract of Chinese medicine is the water extract of ginseng and Cistanche cistanche, and the concentration of the water extract of Chinese medicine is preferably 0.9-1.1g / mL, more preferably 1.0 g / mL, based on the crude drug Cistanche cistanche, the concentration of the Chinese medicine extract is preferably 0.9 to 1.1 g / mL, more preferably 1.0 g / mL. In the present invention, the Chinese medicinal water extract also includes mannose at a concentration of 0.008-0.012 g / mL, more preferably 0.01 g / mL; the function of the mannose is to enhance immunity. In the present invention, the preparation method of the water extract of the traditional Chinese medicine comprises the following steps: soaking the ginseng and cistanche cistanche in water, boiling with a slow fire, decocting and concentrating. In the present invention, the mass ratio of the ginseng and cistanche is preferab...
Embodiment 1
[0034] CpG oligodeoxynucleotide (CpG oligodeoxynucleotide, CpG ODN) aqueous solution, designed and synthesized 478 CpG ODNs (see Table 1 for sequence information), artificially synthesized and dissolved in enzyme-free water to a final concentration of 100ng / μL;
[0035] Water extract of traditional Chinese medicine: take 300g each of ginseng and Cistanche deserticola, wash them, put them in 3000mL deionized water at 25°C for 3h, then decoct them with slow fire for 1h, filter them through a 200-mesh filter and add 3g of mannose to the filtered juice (SIGMA company) , continue to decoct and concentrate to 300mL;
[0036] Serum-free medium Opti-MEM (purchased from GIBCO);
[0037] Liposome: Liposome 2000Plus and Liposome 2000LTX (purchased from Invitrogen)
[0038] Mix 100 μL CpG oligodeoxynucleotide aqueous solution, 5 mL serum-free medium Opti-MEM, 35 μL Liposome 2000Plus, and let stand at 25°C for 5 minutes; add 100 μL Liposome 2000LTX, mix evenly, and let stand for 30 minute...
Embodiment 2
[0052] CpG oligodeoxynucleotide (CpG oligodeoxynucleotide, CpG ODN) aqueous solution, designed and synthesized 478 CpG ODNs (see Table 1 for sequence information), artificially synthesized and dissolved in enzyme-free water to a final concentration of 75ng / μL;
[0053] Water extract of traditional Chinese medicine: take 200g each of ginseng and cistanche, wash, soak in deionized water at 25°C for 3h, decoct in 2800mL deionized water for 1h, add 2.5g mannose (SIGMA company) to the juice, continue decocting and concentrate to 250mL;
[0054] Serum-free medium Opti-MEM (purchased from GIBCO);
[0055] Liposome: Liposome 2000Plus and Liposome 2000LTX (purchased from Invitrogen)
[0056] Mix 80 μL of CpG oligodeoxynucleotide aqueous solution, 4 mL of serum-free medium Opti-MEM, 30 μL of Liposome 2000Plus, and let stand at 25°C for 30 minutes; add 80 μL of Liposome 2000LTX, mix evenly, and let stand for 25 minutes , add 95.81mL Chinese herbal extract, and mix well. Bottle, seal, ...
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