31 results about "Phorbol" patented technology

The invention belongs to the technical field of cell culture, and in particular relates to a method for efficient separation and culture of human primary melanin cells. According to the method provided by the invention, an ROCK inhibitor is added to a culture medium which consists of the following ingredients: a Ham's F-12 medium, dibutiryl cyclic adenosine monophosphate, 3-isobutyl-1-methylxanthine, sodium orthovanadate, phorbol 12-myristinate 13-acetate, fetal calf serum and double-antibody; on the basis of mutual actions of the various ingredients, the melanin cells can achieve cloning growth, so that a culture cycle can be shortened by half and separation efficiency of the melanin cells can be greatly improved; the separation and culture method provided by the invention is simple and easy to operate and is low in amount of medium ingredients; and large-scale production of the human primary melanin cells is achieved.

The invention belongs to the technical field of assisted reproduction, and relates to an oocyte activating solution and application thereof. The oocyte activating solution comprises 1.1 to 1.8 mg/L ofionomycin, 6 to 12 mg/L of calcium ion carrier A23187, 63 to 94 mug/L of puromycin, 19 to 43 mg/L of 6DMAP, 3.4 to 6.6 mg/L of actinone CHX, 28 to 53 mug/L of phorbol PMA, 7.8 to 12.7 mug/L of inositol triphosphate, 1.5 to 2.4 g/L of SrCl2, 16 to 25 mg/L of heparin, and basic culture medium. The oocyte activating solution can effectively activate oocytes, and is suitable for activating the oocytes after ICSI in an in-vitro fertilization assisted reproduction technology.

This invention concerns a process to convert a hydroxyl group (bold in R₃C—OH) in a tigliane-type compound to a hydrogen (bold in R₃C—H) to obtain deoxytigliane-type compounds or structural or functional analogs thereof. The process has wide application particularly to produce specific biologically active compounds in quantity for use as pharmaceuticals. In particular the process can be used to convert phorbol to a 12-deoxytigliane (prostrating which is a therapeutic lead for the treatment of AIDS. New compositions of matter are also disclosed.

The invention belongs to the technical field of the biological medicine, and relates to a microbial conversion method for a euphorbia lathyris diterpene compound. The method comprises the following steps: performing the microbial conversion to euphorbia lathyris phorbol and 7-hydroxy euphorbia lathyris phorbol by using Nocard's bacillus (i) Nocardia sp. (/i) NRRL 5646, and preparing 3-carbonyl euphorbia lathyris phorbol and 7-acetoxyl euphorbia lathyris phorbol derivatives. Proved by the experiment, the polarity of the 3-carbonyl euphorbia lathyris phorbol in the derivative is reduced. Compared with a substrate, the derivative has the better cytotoxic activity. The derivative is an antitumor lead compound with development value.

The invention relates to the field of biological medicines, and relates to a conversion method of a euphorbia lathyris diterpene compound. The method comprises the following steps of utilizing a known microorganism to perform microorganism conversion on the euphorbia lathyris phorbol and a derivative thereof, and combining with chemical acylation, so as to prepare a euphorbia lathyris diterpene derivative compound; especially, utilizing a mucor circinelloides (CICC40242) microorganism to perform hydroxylation biological conversion on the euphorbia lathyris phorbol and euphorbia factor L3, and performing chemical acylation on hydroxyls introduced by C8 and C18, so as to obtain three hydroxylated products and six C8 alcohol esterified products. The method has the advantages that the water solubility of the euphorbia factor L3 is improved by the hydroxylation at site, and the druggability is improved; furthermore, the prepared euphorbia lathyris diterpene compound can be used for preparing anti-tumor activity and anti-tumor multidrug resistance type pharmaceutical compositions. The chemical formula is shown in the specification.

Herein is reported a method for producing a thymocyte supernatant comprising the steps of co-cultivating thymocytes and mononuclear cells at a cell ratio of at least 0.5:1.2 in the presence of phorbol-12-myristate-13-acetate and Phytohemagglutinin M for up to 60 hours, and separating the co-cultivation medium from the cells and thereby producing the thymocyte supernatant.

The invention provides a targeting nanomaterial with a cell membrane coated with Au-Fe3O4 as well as a preparation method and application of the targeting nanomaterial, belonging to the technical field of targeting nanomaterials. The targeting nanomaterial provided by the invention comprises a cancer cell membrane, Au-Fe3O4 nanoparticles, tannic acid and phorbol 12-myristate 13-acetate, wherein the Au-Fe3O4 nanoparticles, the tannic acid and the phorbol 12-myristate 13-acetate are wrapped in the cancer cell membrane. PMA can effectively stimulate cells to generate H2O2 and active oxygen O2 <->, can supplement insufficient H2O2 in cancer cells, and can supplement raw materials for Fenton reaction; the Fe3O4 can be partially decomposed in the presence of the tannic acid to form Fe<2+> and Fe<3+>, Fe<2+> can catalyze H2O2 in the cancer cells to be decomposed into hydroxyl radicals, and TA can promote rapid conversion of Fe<3+> generated by Fe3O4 into Fe<2+>; and meanwhile, Au and Fe3O4 have good photo-thermal conversion efficiency, and can give full play to the therapeutic effect of PTT.

The present invention relates to a diterpene compound derived from Aleurites fordii, and a pharmaceutical composition for treating or preventing viral infectious diseases, a health functional food for preventing or ameliorating viral infectious diseases and a composition for enhancing the production of interferon-gamma, which comprise the diterpene compound. Furthermore, the present invention relates to a method for preventing or treating viral infectious diseases including administering the composition to a subject having a viral infectious disease occurrence or a risk thereof with a therapeutically effective dose.

The invention discloses a method for evaluating the immunomodulatory function of a substance, which comprises the following steps: preparing a primary natural immune cell suspension from the spleen of an acquired immune cell deficient mouse; culturing the immune cells and OVA to obtain a culture system; collecting spleen and lymph node single-cell suspensions of a T cell receptor transgenic mouse, and carrying out magnetic bead sorting to obtain CD4 + CD62L + Naive T cells; adding the CD4 + CD62L + Naive T cells into the culture system, and adding a substance to be evaluated into the culture system at the concentration of 100-500mu g/mL in the culture process; collecting all cells, performing stimulation culture on the cells by using phorbol and ionomycin, performing membrane rupture, matching and fixing, adding a fluorescent antibody, and detecting the proportion of Th1, Th2 or Treg by using a flow cytometer so as to evaluate the immunomodulatory function of the substance. The method can accurately evaluate whether the active substance has the immune regulation function, and is low in cost and short in period.

The invention provides an extraction method for phorbol to overcome the problems that conventional silica gel column chromatography is employed for phorbol extraction in the prior art and consumes a large amount of solvents and conventional extraction processes are complicated and have low purification efficiency. The extraction method for phorbol in the invention comprises the following steps: (1) weighing the seeds of Jatropha carcass and carrying out physical pulverization; (2) carrying out refluxing with an ethanol solution, then carrying out extraction, and then concentrating the obtainedextract until the extract is dry; (3) extracting a product concentrated to dryness in the step (2) with ethyl acetate and then carrying out concentration to dryness; (4) weighing a concentrated product obtained in the step (3), adding a sodium hydroxide solution and carrying out stirring until TLC shows that alkaline hydrolysis is completed; and (5) subjecting a crystallization product obtained in the step (4) to recrystallization with absolute ethanol so as to obtain a white needle crystal.

The invention relates to a construction method and a use of a nanometer targeting preparation of a micro-molecular immune drug. Ingenol phorbol angelate which cannot be dissolved in water and only can be administrated in a skin smearing manner as a gel, and its derivatives are loaded to a carrier auxiliary material to form nanometer particles, and then the nanometer particles are lyophilized to form a powder injection, so the water solubility problem is solved, the stability of the active components of the micro-molecular immune drug is improved, the problem of no vein administration or tumor interventional administration due to too high hydrophobicity and toxicity is solved, and the use of the micro-molecular immune drug is expanded.

The invention relates to the technical field of natural medicine extraction, and particularly discloses a method for extracting phorbol from croton oil by using a Girard's reagent. The method comprises the following steps: S1, mixing 1 part of croton oil with 5-25 parts of a methanol saturated solution of a Girard's reagent according to a feed liquid volume ratio, and carrying out a stirring reaction at 18-50 DEG C for 30-180 min to obtain a reaction solution; S2, concentrating a reaction solution; S3, extracting and removing croton oil by using petroleum ether to obtain a water phase containing the phorbol Girard's reagent salt; S4, adjusting the pH value to 2-3, and extracting the phorbol in the water phase by using petroleum ether; S5, concentrating the petroleum ether phase to obtain an oily substance; and S6, carrying out alcoholysis, acidification and filtration on the oily substance to obtain a crude product of the phorbol. According to the method, carbonyl reacts with a Girard's reagent, and then a large amount of impurities can be removed by an extraction method due to different polarities of the components, so that a crude product of the phorbol with relatively high purity is obtained. The method has the characteristics of simplicity and convenience in operation, short time consumption and low solvent consumption, and is easy to popularize and apply industrially.

The invention relates to batch half-channel function level detection method based on fluorescent dye uptak. The method comprises the steps: (1) suspension culture: taking THP-1 cells of a suspension culture cell line, using an RPMI1640 culture solution added with beta-mercaptoethanol, and regularly adding or replacing the culture solution to ensure the steady state of the living environment of the cells; monitoring that the concentration of the cell suspension does not exceed 1 * 10 < 6 >/ml; adjusting the concentration of the cell suspension to be 4 * 10 < 5 >/ml, adding tetradecanoyl phorbol acetate, inoculating the cell suspension to a 96-well plate, and changing the solution 24 hours later to remove non-adherent cells; using a blank RPMI1640 culture solution for maintaining culture so as to remove the action effect of tetradecanoyl phorbol acetate; and (2) taking in a fluorescent dye to detect the activity of the semi-channel, taking ethidium bromide as the fluorescent dye, preparing an ethidium bromide fluorescent staining stock solution by using a standard RPMI1640 culture solution, adding the ethidium bromide fluorescent staining stock solution into a 96-well plate, adding the fluorescent staining solution, placing the 96-well plate on ice, incubating in a dark place, and placing the 96-well plate into a fluorescence microplate reader to detect the fluorescence intensity result.

The invention relates to the technical field of natural medicine extraction, and particularly discloses a method for separating phorbol by utilizing an ultrasonic-assisted alcoholysis-extraction coupling technology. The method comprises the following steps: S1, uniformly mixing 1 part of croton oil with 5-25 parts of a saturated alkaline alcoholic solution according to a volume ratio of feed liquid, carrying out alcoholysis-extraction under the action of ultrasonic waves, and carrying out standing separation to obtain an alkaline supernatant containing phorbol and a croton oil lower phase liquid; and S2, adjusting the pH value of the obtained alkaline supernatant to be neutral by using an acidic alcohol solution, and removing the obtained precipitate to obtain a neutral alcohol solution containing the phorbol. The ultrasonic-assisted alcoholysis-extraction coupling technology is used for extracting the phorbol from the croton oil, alcoholysis and extraction are carried out synchronously, and alcoholysis-extraction coupling is carried out, so the alcoholysis efficiency is improved advantageously, alcoholysis-extraction is fast and efficient under the assistance of ultrasonic waves,time is saved, and the purity of the phorbol is improved. The method has the advantages of few operation steps, short time consumption, low solvent consumption and easiness in promotion and application.