140 results about "Mtt method" patented technology

The invention discloses a type of amino acid modified curcumin, a synthesis method thereof, and an application thereof. According to the invention, structural modification is carried out upon curcumin by using natural amino acid, such that 1-(4-hydroxy-3-methoxyphenyl)-7-(4-oxyacetyl amino carbobenzoxy-3-methoxyphenyl)-1,6-heptadiene-3,5-diketone which is represented by a general formula (I) is obtained. With an MTT method, evaluation is carried out upon the compound represented by the general formula (I) in the inhabitance of proliferation activities of four cells which are K562, H22, HL60 and S180. IC50 values of the compound represented by the general formula (I) in inhibiting the proliferations of the four tumor cells are calculated. As a result of experiments, with the compound represented by the general formula (I), proliferations of tumor cells can be substantially inhibited. The compound has excellent anti-tumor activity, and can be prepared into anti-tumor medicines.

A process for detecting the sensitivity of tumor cell to medicine includes such steps as adding the antineoplastic medicine in the holes on detecting plate, freeze solidifying, adding tumor cells to the holes, reducing it to color deposit by MTT method, preparing color solution, enzyme-linked assay, calculation and comparing to obtain sensitivity value, and judging. Its advantages are simple operation, and high correctness and stability.

The invention relates to a marine bacillus and its polypeptide with antitumor activity. The strain is preserved at China Center for Type Culture Collection in Wuhan, and its preservation number is CCTCC M 2013063. A novel marine bacillus polypeptide with anti-tumor activity is obtained from a fermentation product of the strain. By means of an MTT method, the invention finds that the polypeptide has an obvious proliferation inhibition effect on human hepatoma carcinoma cell BEL-7402, human breast cancer cell MCF-7, human glioma cell U251, human non-small-cell lung cancer cell A549 and other tumor cells, has relatively small cytotoxicity on human fibroblast HFL1, and can be applied in drugs treating human liver cancer, gliomas, lung cancer and breast cancer.

A medicine combination has the function of resisting lung cancer. The combination is mixed with rhizoma paridis saponin and milk vetch root amylose according to the weight ratio of 3 to 1. The rhizoma paridis is extracted by alcohol and then the rhizoma paridis extract is obtained through gradient elutriation of macroporous absorption resin alcohol; the milk vetch root is extracted by water and then the protein is settled and removed to get the milk vetch root amylose; the two extracts are mixed and the medicine is made. The MTT activities experiment in vitro with the MTT method has proven that the medicine combination can obviously control the growth of various lung cancer cells such as LA795 lung adenocarcinoma cell and the IC50 can reach 26.73 ug / ml; the experiment of mouse with the lung cancer tumor has represented that the tumor constraint rate can reach 55.63 percent; in this way, lung transfer of hypodermic transplanted tumor of the mouse with the lung cancer tumor can be obviously controlled and the tumor cells can be brought to death; moreover, spleen index and thymus index can be promoted and the medicine is innocuous and has no side effect. In addition, compared with the raw materials, the medicine combination has high activity and clear function; moreover, the medicine can be made into different types.

The invention relates to an MTT (thiazolyl blue) cell toxicity test method of biological assessment of a total particle matter in cigarette smoke and belongs to the technical field of safety assessment of tobacco and cigarette smoke. The MTT cell toxicity test method is characterized by comprising the following steps of: inoculating and culturing immortalized human bronchial epithelial cells (BEAS-2B cells) and contaminating the total particle matter in the cigarette smoke; detecting the cell survival rate by adopting an MTT method; and analyzing and evaluating the cell toxicity of the total particle matter in the cigarette smoke according to a test result. Compared with the prior art, the MTT cell toxicity test method has the following characteristics that BEAS-2B cells are human cells and are target organ source cells acting on a human body by the smoke; a BEAS-2B cell system is used for carrying out cell toxicity assessment on the cigarette smoke and has the strong pertinence; in a testing process, steps of washing the cells for a plurality of times are reduced; and a formaldehyde fixing step does not need to be carried out so that a testing period is shortened, the operation is rapid and convenient, the sensitivity is high and the result is stable and reliable. Moreover, the MTT test method disclosed by the invention is further applicable to a smoke cell toxicity test of various cell systems and the commonality is strong.

The invention discloses application of Lunasin polypeptide in the aspect of preparing substance with weight-reducing activity. Lunasin is substance with weight-reducing activity and is prepared by inhibiting differentiation of mouse preadipocyte (3T3-L1) through a PPAR gamma passage. The application includes: using an MTT method to detect influence, on 3T3-L1 adipocyte activity, of Lunasin at different dosages; detecting inhibiting effect, on adipocyte differentiation, of Lunasin at different dosages. Compared with current study conditions of Lunasin, the application has the advantages that the application shows that Lunasin can inhibit differentiation of insulin-induced 3T3-L1 cells, reduce accumulation of lipid droplets in lipid and inhibit mRNA expression of PPAR gamma, Lunasin presents dosage dependence, and effect of Lunasin with dosage higher than 50ug / mL is better than that of rosiglitazone of 5ug / mL; the application shows that Lunasin has potential weight-reducing effect.

The invention relates to assessment and fermentation of marine actinomycete WBF9 that is used for producing anti-tumor compound and the separation and purification of the anti-tumor compound and the cytotoxic activity, and pertains to the microscopic organisms medicine filed. Bacterial strain WBF9, which is streptomyces griseorubens that is separated from the sea for the first time, is identified as one of the new bacterial strains of streptomyces griseorubens by traditionally classification and 16SrDNA genetic analysis. Culture medium (glucose: 1 percent, potato: 15 percent, yeast powder: 0.6 percent, calcium chloride: 0.12 percent and natural seawater, pH value 7.2) is applied by the bacterial strain for fermentation, and the compound extracted from the fermentation liquor, which has strong anti-tumor activity and identified as Norharmane by MS and NMR technique, is the first compound that is acquired by separating marine actinomycete. According to the MTT method, the compound is found to have cytotoxicity against four tumor cells: KB, BGC 803, Hep G2 and SGC 7901, and is a potential anti-tumor medicine.

The invention discloses a fluorescence-labeled magnetic kaempferol microsphere system and a preparation method thereof. The preparation method comprises the following steps: (1) firstly, dissolving a hydrophobic drug, namely hydrophobic drug, in an oil-base modified magnetic fluid, uniformly mixing, dropping the dissolved kaempferol to a proper amount of a bovine serum albumin solution by virtue of an injector, and synthesizing a magnetic kaempferol protein microsphere through an ultrasonic chemical method; (2) then, connecting to self-made graphene quantum dots through an amino carboxylic reaction so as to obtain a fluorescent and magnetic dual-functional kaempferol microsphere; and (3) co-cultivating the a fluorescent and magnetic dual-functional kaempferol microsphere with human Hela cells, and detecting cytotoxicity by virtue of an MTT method. The fluorescence-labeled magnetic kaempferol microsphere disclosed by the invention is low in preparation cost and high in drug loading rate; the microsphere not only has the superparamagnetism of magnetic particles but also reserves the excellent fluorescence performance of the graphene quantum dots; and the microsphere can achieve an excellent effect on medicines for tumor targeted therapy.

The invention provides a preparation method and application of a Tf (transferrin) modified nano diamond targeted medicine. The preparation method of the nano diamond targeted medicine comprises the following steps: dissolving transferrin Tf into a PBS (phosphate buffer solution), then adding DOX (doxorubicin) and reacting to obtain a Tf-DOX compound; and carrying out reaction on carboxylated nano diamond and the Tf-DOX compound to obtain the nano diamond targeted medicine ND-(Tf-DOX). The nano diamond targeted medicine acts on HepG2, and treatment on cells by virtue of low temperature, different inhibitors and free Tf shows that the nano diamond targeted medicine has energy dependence; with normal human HEK293 as reference, study on absorption of the cells for the nano diamond targeted medicine shows that the nano diamond targeted medicine has targeted selectivity; with human HepG2 and human HeLa as references, detection on cytotoxicity of the nano diamond targeted medicine by adopting an MTT method shows that the nano diamond targeted medicine has a targeted anti-tumour effect and selectivity for cancer cells; and study on a tumour inhibiting effect of the nano diamond targeted medicine in vivo by taking tumour-bearing mice as models shows that the nano diamond targeted medicine can be used for inhibiting tumours and has low toxic and side effects on DOX. Therefore, the nano diamond targeted medicine can be applied to preparation of an anti-tumour medicine.

The invention discloses a screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia. The tea polyphenol in tea and golden camellia is extracted and prepared from different types of tea and golden camellia, and the tea polyphenol acts on human esophageal squamous cells and adenocarcinoma cell OE33 which are cultured in vitro; an anti-proliferative effect on esophagus cancer by the tea polyphenol is detected in an MTT method, and an inducing differentiation effect on the esophagus cancer by the tea polyphenol is detected by flow cytometry so as to observe cell cycles and apoptotic effects; cell morphological change is observed by using a fluorescence inversion microscope; protein expression change in the esophagus cancer cells is analyzed by means of immunohisochemistry, Real-time fluorescent quantitation RT-PCR and Westem-blot. Based on SPSS1 statistics, the detection results are analyzed, and differential expression proteins obtained by performing action on the esophagus cancer by the tea polyphenol in the tea and golden camellia are detected, and associated proteins are screened out according to database search and matching. In the invention, a mechanism of the function of tea polyphenol can be expounded in accordance with the research on marker molecules of the differential expression proteins acted by the tea polyphenol.

The invention relates to a method for verifying and fermenting polar marine ray fungi AFN1007 producing antitumor active substance, and the research on the antitumor activity of the zymotic fluid thereof, belonging to the microbial medicine field. The strain AFN1007 is verified to be a novel kind of Streptomyces by the traditional classification and the analysis of 16S rDNA gene of the AFN1007. The compositions of a culture medium by percentage are: 1 percent of soybean flour, 1 percent of tragantine, 0.1 percent of KNO3, 0.05 percent of MgSO4, 0.001 percent of FeSO4, 0.05 percent of NaCl and natural seawater with a pH value between 7.2 and 7.4. The strain is fermented by using the culture medium. The MTT method shows that the zymotic fluid has strong antitumor action on four tumor cells of KB, BGC 803, Hep G2 and Hela and an antitumor compound with a novel structure is expected to be found in the zymotic fluid.

The invention aims at establishing a method for quickly measuring total number of live bacteria of luminous bacteria, in particular to a method for counting the live bacteria based on the MTT coloration principle. The invention adopts MTT colorimetric method to measure the viable count of the luminous bacteria and carries out optimizing selection on the detection condition of the MTT colorimetric method. Therefore, the optimal method for measuring total number of the live bacteria of the luminous bacteria is determined. In the invention, first, the product of formazan generated from the reaction of succinate dehydrogenase in MTT and live thalli is dissolved in DMSO, light absorption value is determined within the range of 400nm-800nm, and the largest detected absorption wavelength is determined to be 500nm. By optimizing time and influencing the light absorption value by temperature, the optimal measurement condition is detected: the reaction time is 2h, and the reaction temperature is 25 DEG C. The invention has novel detection principle, simple and fast detection method, good repeatability, can be used for the quantitative detection of the number of the live bacteria of the luminous bacteria in the fields of food hygiene and safety, environmental monitoring and the like. The invention can also be applied to the tests of microbiology, immunology and other disciplines and used for measurement of the activity and number of the live bacteria materials. The invention can also develop the method for measuring the total number of the bacteria which is used for replacing the traditional method for measuring the total number of the live bacteria.

The invention discloses a sesquiterpene quinone compound separated and purified from abelmoschus sagittifolius, as well as a preparation method and applications thereof, relating to the technical field of medicines. The new sesquiterpene quinone compound with antitumor activity is named as Acylhisbiscone B, with the molecular formula of C17H22O4 and the structural formula shown in the specification. According to the preparation method, the detection on the antitumor activity of the new compound is carried out by adopting an MTT method through combination of column chromatography and preparative liquid chromatography purification methods. In vitro experiment results show that the new sesquiterpene quinone compound has activity on significantly inhibiting the growth of Hela (human cervical carcinoma cells) and HepG-2 (human hepatoma carcinoma cells). The new sesquiterpene quinone compound can provide new chemical entity or lead compound for research of antitumor medicaments, and can also be used as food raw materials for preparation of health food.

The invention discloses application of conyza blinii total saponins for preparing an anti-neoplastic drug. An MTT method is used for detecting a growth inhibition ratio of the conyza blinii total saponins for cervical carcinoma Hela cells and lung carcinoma SPC-A1 cells and calculating IC50 (50 percent of Inhibiting Concentration); a result explains that the conyza blinii total saponins has time and concentration dependency for the growth inhibition of the Hela cells and the SPC-A1 cells; an agarose gel electrophoresis method is adopted to detect acted cells of the conyza blinii total saponins in order to display an apoptotic characteristic DNA ladder and indicate that the conyza blinii total saponins has an apoptotic function for the Hela cells and the SPC-A1 cells; and a Western-blot method is adopted to detect the influence on apoptotic key protein caspase-3 content by the conyza blinii total saponins, obtain that after the conyza blinii total saponins acts on the cells, casepase-3 is increased through a semiquantitative analysis and indicate that the growth inhibition of the cells by the conyza blinii total saponins is regulated by an apoptosis mechanism.

The invention relates to the application of the compound metabolized of fungus between kidney beans. The analysis would take X-Ray, MS and biological activity testing processes. According to the test of biological testing of compound Mycoepoxydiene, using MTT method to testing the anti-tumor activity, the human oral cavity cancer KB cell is IC50<6.25ug / ml, and the human osteosarcoma MG63 cell is IC50=34.6ug / ml. The compound Mycoepoxydiene has anti-tumor activity, and could be used in anti-tumor medicine or the primer of other biology activity.

The invention relates to the technical field of microbes, in particular to a microbe bacterial strain in which a new compound can be obtained by separating in the fermentation liquor. The invention is named as Actinopolyspora erythraea YIM 90600T, and is preserved in a depositary institution appointed by the State Intellectual Property OfficeState Intellectual Property Office with the preserving number of CCTCC M208243. The Actinopolyspora erythraea of the invention is separated from saline soil in AydingkolHu in Xinjiang of China. The depositary institution appointed by the State Intellectual Property OfficeState Intellectual Property Office is the China Center for Type Culture Collection in Wuhan University in China. The postcode is 430072, and the preserving data is Oct. 9, 2007. A new erythrocin analogue having antibacterial tumour cytoactive (MTT method) can be obtained by separating in the fermentation liquor of the Actinopolyspora erythraea of the invention can separate.

The invention discloses a preparation method of a magnetic graphene drug-loading system. The preparation method comprises the following steps of: (1) modifying oxidized graphene by using two types of surface active agents, and synthesizing into magnetic oxidized graphene by a hydrothermal synthesis method; (2) modifying by an amphiphilic polymer PF127, reducing the magnetic oxidized graphene into magnetic graphene with good biocompatibility by hydrazine hydrate; (3) loading a hydrophobic anti-cancer drug (paclitaxel) onto the magnetic graphene by pi-pi stacking effect and preparing into the magnetic graphene drug-loading system with integration of an anti-cancer effect, imaging and biocompatibility; (4) culturing the magnetic graphene drug-loading system and MCF-7 breast cancer cells together, and detecting the cell toxicity by an MTT method. The invention discloses application of the magnetic graphene drug-loading system in treating the breast cancer. The magnetic graphene drug-loading system disclosed by the invention is good in drug dispersity, strong in magnetic responsibility, good in biocompatibility and high in drug loading amount, and can show better inhibiting effect for MCF-7 breast cancer cells under the condition of less use amount (200mug / mL).

The invention discloses a polyoxometalate compound modified by amino acid.The compound has the general formula of Am[B3XMo6O21].nH2O and is prepared through the steps of dissolving Na2MoO4.H2O, Se, As, Sb or Bi oxide, glycine, alanine, histidine, lysine, tryptophan, phenylalanine, methionine, threonine, isoleucine, leucine or arginine and KCl in H2O, putting the solution on a magnetic stirrer to be mixed and stirred, adjusting pH of the solution to be 3.4 to 3.6 through HCl, stirring and heating reflux for 0.5-2 hours, conducting cooling and filtering, standing for 1-2 days, and collecting crystals.It is determined through the MTT method that the IC50 value of HL-60 cells is 0.127 microM, the IC50 value of U937 cells is 2.731 microM, the IC50 value of U937 cells is 2.731 microM, and the IC50 value of HUVECs cells is 889.18 microM.The polyoxometalate compound is high in leukemia resistant activity, low in toxicity and low in price.

The invention relates to the technical field of medicine preparation, and in particular relates to application of 3,5,3',4'-trihydroxy-stilbene-3'-b-D-glucoside in preparation of medicines for treating cancers. The cytotoxic effect of the 3,5,3',4'-trihydroxy-stilbene-3'-b-D-glucoside on 12 human tumor cells cultured in vitro and a human normal cell is tested through an MTT method; and the 3,5,3',4'-trihydroxy-stilbene-3'-b-D-glucoside has a relatively good inhibiting effect on proliferation of a human lung cancer cell line, a human colon cancer cell line, a human erythroleukemia cell line, a human gastric cancer cell line and a human breast cancer cell line, and has no inhibiting action on proliferation of human normal cells. The curative effect of the 3,5,3',4'-trihydroxy-stilbene-3'-b-D-glucoside on a nude mouse transplantation tumor of a human lung cancer cell line A549 is also tested. The result shows that the 3,5,3',4'-trihydroxy-stilbene-3'-b-D-glucoside has obvious inhibiting action on proliferation of the nude mouse transplantation tumor of the human lung cancer cell line A549 after continuous intravenous injection for 14 days; and the dose-effect relationship is obvious.

The invention discloses the use of anthocyanin in medicines for treating atherosclerosis and the application for preventing and treating atherosclerosis with regulation and control of CHOP gene, and relates to the improvement effect of anthocyanin monomer cyanidin-3, 5-diglucoside on macrophage injuries caused by oxidized low density lipoprotein, and discusses the mechanism of action thereof. In the invention, mouse macrophage cell line RAW264.7 is tested, and the effect of cyanidin-3, 5-diglucoside on activity of RAW264.7 macrophage processed by the oxidized low density lipoprotein is determined based on MTT method, the total RNA of cells is extracted, Affymetrix Mouse U4302.0 gene expression profile chip is applied to carry out marking hybridization experiments, differential gene analysis is carried out for the scanning results of Affyemtrix gene chip with MTT method, and the differential genes are screened, and further real-time quantitative PRC method is utilized to further validate the expressed differential gene, The validation results verify that the cyanidin-3, 5-diglucoside can regulate CHOP gene expression down, and has obvious protective action on macrophages injured by oxidized low density lipoprotein.

The invention discloses an oyster polypeptide-nano selenium granule compound having hangover-relieving and liver-protecting effects and a preparation method and application thereof. The method includes: homogenizing oyster meat, adding water and active enzyme, enzymolyzing, allowing boiling water bath, centrifuging to take supernate, cold-drying the supernate to obtain oyster polypeptide powder, adding the oyster polypeptide powder into water, well mixing to obtain a solution, adding a sodium arsenite solution into the oyster polysaccharide solution, well mixing, adding an ascorbic acid solution while stirring, allowing water bath reaction, dialyzing to obtain functionalized nano selenium sol, ultrasonically treating, and freeze-drying to obtain the oyster polypeptide-nano selenium granulecompound. Oyster polypeptide is used to stabilize nano selenium, so that stability of nano selenium is remarkably improved, and nano selenium can be stably stored at 4 DEG C for one month; through MTT method detection, nano granules can relieve damage effect of alcohol or hydrogen peroxide on liver. If the compound is converted into healthcare products having a liver protecting function and capable of supplementing selenium element, great social and economic benefit can be generated.

The present invention provides a method for isolating and purifying immuno-modulating polypeptide from cow placenta, which is characterized by using the steps of anion-exchange chromatography, gel exclusion chromatography and reverse-phase high performance liquid chromatography to isolate and purify immuno-modulating polypeptide from cow placenta, identifying its activity of stimulating lymphocyte proliferation in vitro by MTT method, then determining its molecular weight by MALDI-TOF-MS, its isoelectric point by CIEF and its amino acid sequence with analyzer for protein sequencing. Since the obtained immuno-modulating polypeptide by the method according to the present invention has more than 90% purity, its bioactivity can reach medicinal standards.

The invention relates to clam active polypeptide, in particular to application of clam active polypeptide in preparing an anti-lung cancer medicine. The invention provides the application of the clam active polypeptide in preparing the anti-lung cancer medicine. As shown by experimental results by an MTT method and a cell counting method, the clam polypeptide has specific effect of inducing and killing lung cancer cells A549, and has obvious effect of inhibiting the lung cancer cells A549, so that the clam active polypeptide can be used for preparing the anti-lung cancer medicine.

The application discloses estrogen effects of a Ganoderma lucidum component GL-1 and preliminary discussion of an action mechanism thereof. A computer-assisted simulation computation is used for computing binding capability of GL-1 and an estrogen acceptor, a MTT method is used for detecting influences of GL-1 on proliferation of MCF-7 cells and MDA-MB-231 cells, influences of GL-1 on proliferation of MCF-7 cells induced by estrogen as well as influences of an estrogen acceptor inhibitor ICI182,780 on proliferation of MCF-7 cells induced by GL-1; Western blot is used for detecting expression conditions of ER alpha and ER beta monoclonal antibodies. Results show that the Ganoderma lucidum component GL-1 and an estrogen acceptor beta have good combination capability, and Ganoderma lucidum component GL-1 has estrogen-like effects, can be combined with the estrogen acceptor, and influences estrogen secretion and ER beta expression.

The invention relates to a preparation method for Sinkiang turnip polysaccharide having immunoregulatory activity. The method includes the steps that Sinkiang turnip root tubers are dried and smashed, ethyl alcohol with the concentration of 80% is used for removing impurities from the Sinkiang turnip root tubers, the Sinkiang turnip root tubers are subjected to hot water extraction, suction filtration is conducted to extract supernate, absolute ethyl alcohol precipitation and centrifugation are conducted, precipitate is washed sequentially through diethyl ether, ethyl alcohol and acetone and is dried until the constant weight state is reached, and the Sinkiang turnip polysaccharide is obtained. An MTT method is used for analyzing the immunoregulatory activity of the Sinkiang turnip polysaccharide and it is discovered that the Sinkiang turnip polysaccharide has a remarkable effect of promoting mouse peritoneal macrophage proliferation. The preparation method for the Sinkiang turnip polysaccharide has the advantages that the extraction yield is high, cost is low, the structure of the polysaccharide is complete, and no pollution is caused. The prepared Sinkiang turnip polysaccharide has the prominent immunoregulatory activity, thereby having broad application prospects in the food industry and the pharmaceutical industry.

The invention belongs to the technical field of nanomechanics and molecular biology, and relates to preparation of a carbon quantum dot fluorescent probe and application of the carbon quantum dot fluorescent probe in selective detection of active oxygen. Carbon quantum dots with specific detection functions on different active oxygen can be prepared by the method. A series of different carbon sources are used as precursors, and different carbon quantum dots are synthesized through a one-step hydrothermal reaction. XPS characterization shows that the types and contents of oxygen-containing groups and nitrogen-containing groups on the surfaces of the carbon quantum dots generated by different carbon sources are different, and various groups have different binding capacities with active oxygen such as superoxide anions (O2<.->) and hydroxyl radicals (.OH), so that the carbon quantum dots can be used for specific detection of related active oxygen. Meanwhile, an MTT method proves that thecarbon quantum dot synthesized by the method has good biocompatibility, can be used for in-vivo cell imaging and online detection, detects the content of active oxygen generated by stimulating livingcells by a drug, and is used for related cell counting.

A establishment method of a novel high-throughput screening model is provided in the invention, which aims to solve the problems of nondeterminacy and incorrespondence exsiting in general in vitro high-throughput screening method. The model provided by the invention furthest simulates animal physiological environment, thus the accuracy of drug screening is increased and the results obtained by the method of in-vitro screening are closer to that of clinical application. The technical scheme of the present invention is as follows: Animal medicated serum is obtained by absorpting and separating drug active ingredients during the process of in vivo circulation of animals, thereby the medicated serum is added to cell culture media or maintenance media after drug is subject to the process of in vivo metabolism of animals. Cell activity is calculated by utilizing micro cell culture method, cytopathic effects (CPE) measurement and MTT method.

The present invention relates to technical field of medicine, and relates to the use of diterperoid compound glaucocalyxin X in isodon japonica (Burm.f) Hara var.glaucocalyx (Maxim.) Hara in preparing anti-cancer medicine. The invention adopts an MTT method for executing cytotoxicity experiment. The result shows that the glaucocalyxin X equally has remarkable cytotoxicity and therefore can be used for preparing anti-cancer medicine. The invention provides a new source for preparing the anti-cancer medicine.