710 results about "IC50" patented technology

The invention relates to an aflatoxin nano antibody gene pool, a construction method and application of the aflatoxin nano antibody gene pool as well as an aflatoxin B1 nano antibody 2014AFB-G15. The aflatoxin nano antibody gene pool is prepared by extracting RNA (ribonucleic acid) in alpaca blood after immunization of an aflatoxin B1 antigen, performing specific amplification on a variable region gene of an alpaca heavy chain antibody by adopting an RT-PCR (reverse transcription-polymerase chain reaction) method to obtain an aflatoxin nano antibody VHH gene, and then performing transformation after connecting with a pCANTAB5E (his) vector. The aflatoxin B1 nano antibody 2014AFB-G15 obtained by screening, disclosed by the invention, has the characteristics of organic reagent resistance, high temperature resistance and the like, and is good in stability; the IC50 (half maximal inhibitory concentration) of the aflatoxin B1 nano antibody 2014AFB-G15 to the aflatoxin B1 is 0.66ng / mL, and the cross reactivity of the aflatoxin B1 nano antibody 2014AFB-G15 to the aflatoxins B2, G1,G2 and M1 is 22.6%, 0.95%, 32.1% and 26% respectively.

The invention provides a hybridoma cell line 1C11 and an anti-aflatoxin general monoclonal antibody secreted by the same as well as the applications thereof. The hybridoma cell line 1C11 can be used for preparing a high-titer aflatoxin antibody, and a mouse hydroperitoneum antibody is measured to reach 5.12*106 by using an ELISA (Enzyme-Linked Immunosorbent Assay). The anti-aflatoxin general monoclonal antibody has high sensitivity, respectively reaches the IC50 (50% inhibiting concentration) of aflatoxin B1, B2, G1 and G2 to be 1.2, 1.3, 2.2 and 18.0 pg / mL, is the antibody with highest sensitivity among currently reported four aflatoxin antibodies, is used for measuring the total aflatoxin amounts, i.e. the total amounts of the aflatoxin B1, B2, G1 and G2 and has great practical application values.

Disclosed is a novel anti-inflammatory pharmaceutical composition that exhibits potent and selective inhibition of the cycloooxygenase-2 (COX-2) enzyme. The formulation consists of a hops extract that exhibits COX-2 selectivity as defined by dividing the IC50 COX-2 / IC50COX-1 concentrations that are determined by testing with the William Harvey Whole Blood Assay (WHMA), and falls in the range of 0.011 to 0.2. Such compositions may also optionally contain high levels of alpha acids and low levels of beta acids, some flavonoid compounds, and virtually no essential oils. Such compositions are useful for treating conditions that manifest as inflammatory pain, or are impacted by the COX-2 enzyme. The recited compositions are particularly beneficial for treating osteoarthritis and rheumatoid arthritis, and can be used for chronic pain with reduced gastric side-effects.

The invention relates to novel application of berberine and derivatives thereof in preparation of medicines, in particular to application of berberine and derivatives thereof in preparation of indole amine 2, 3-dioxygenase IDO inhibitor, belonging to the medicine field. According to IDO inhibition activity detection, reversible inhibition judgment, inhibitor type judgment, Ki value determination and median effective inhibition concentration IC50 determination, the results show that berberine is reversible inhibitor and inhibition constant Ki is 8muM; and jatrorrhizine hydrochloride and palmatine hydrochloride are irreversible inhibitors, and the median effective inhibition concentrations IC50 thereof are respectively 123muM and 126muM. When the berberine and the derivatives thereof disclosed in the invention are used as IDO inhibitors, the application prospect is wide and the IDO inhibitors can be used for treating serious diseases such as cancers, AIDS, Alzheimer diseases, tristimania, cataract and the like with pathological feature of IDO-mediated tryptophan metabolism.

The invention belongs to the field of medicine, and relates to D-configuration polypeptide with high stability and capable of realizing mediated targeting of acetylcholine receptor high-expression cells and crossing corresponding barrier membranes and a nano drug delivery system thereof as well as an application in in-vivo and in-vitro brain targeting and in treatment of brain diseases and the like. Test results indicate that DCDX and the acetylcholine receptor are combined with IC50 to obtain 84.5nM which is stable in serum and tolerates hydrolysis of protease; the model drug carried by DCDX is specifically taken in by the positive cells expressing the acetylcholine receptor and has an ability of crossing the barrier formed by the kind of cells; and the nano drug delivery system made of a DCDX-modified polymer carrier material can deliver the entrapped model drug to the target tissue while the drug effect is remarkably improved. The D-configuration polypeptide DCDX provided by the invention can mediate active targeting of the drug or nano drug delivery system and has a good application prospect in the diagnosis and treatment of multiple diseases.

The invention provides a compound with the structure shown in a formula I, a preparation method of the compound, and an application of the compound as a tyrosine kinase and / or serine-threonine kinase inhibitor. The compound has a good inhibitory action on the activity of multiple kinases, and has an obvious inhibitory effect on tyrosine kinase and / or serine-threonine kinase in biochemical level and cell level in vitro (P is less than 0.05), and the half inhibitory concentration (IC50) for c-Met kinase is commonly below 10<-6>mol / L; and the compound also has an obvious inhibitory effect on proliferation of multiple tumor cells (P is less than 0.05), and the IC50 is respectively below 10<-5>mol / L. The compound with the structure shown in the formula I can be applied to preparing medicines for treating diseases related to protein kinase in organisms. The formula I is shown in the specification.

The invention discloses a method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides. The method comprises the following particular steps that the activated rhodobacter sphaeroides is inoculated to an MMS (methylmercuric sulfate) culture medium; aerobic culture is performed until OD600 (optical density 600) is 0.6-0.8; a culture condition is adjusted to be microaerobic; the carotenoid is induced to be quickly and greatly synthesized; and then extraction and purification are performed. The time for producing the carotenoid by the method is short and only 36h; the yield of thallus is up to 6g / L; the yield and the productivity of the carotenoid are 8.287mg / L and 1911mug / g respectively; the carotenoid has good capacity for removing DPPH (1,1-diphenyl-2-picryl-hydrazyl) radicals; and IC50 (half maximal inhibitory concentration) is about 8mug / mL.

The invention provides a hybridoma cell strain 2D3, a monoclonal antibody to zearalenone secreted by the hybridoma cell strain 2D3 and application of the monoclonal antibody. The hybridoma cell strain 2D3 is preserved in China Center for Type Culture Collection with an accession number of CCTCC No. C201328 and can be used for preparation of a high-titer monoclonal antibody to zearalenone. According to detection results of enzyme linked immunosorbent assay (ELISA), the titer of the monoclonal antibody to zearalenone prepared through purification of mouse ascites can reach 1.5 * 10<5>. The monoclonal antibody to zearalenone has high sensitivity, half maximal inhibitory concentration IC50 of 20 pg / mL to zearalenone and cross reactivity of 4.9%, 3.3% and 3.2% with beta-zearalanel, alpha-zearalanel and beta-zeranol, respectively. The monoclonal antibody to zearalenone can be used for determination of the content of zearalenone.

A synthetic method of a general artificial antigen of phthalate plasticizers for immunodetection belongs to the field of bio-chemical engineering technology. The synthetic method of the present invention comprises the following steps of carrying out an esterification reaction between phthalic acid and 6-(fluorenyl methoxy carbonyl acyl-amino)-1-hexanol, removing fluorene methoxy carbonyl acyl protecting groups to form a hapten, and coupling with amino from carrier protein to obtain the general artificial antigen of phthalate plasticizers. Experimental results disclose that titer of antiserum obtained by immunizing animals by the antigen of the invention is up to 160000;the 50% inhibiting concentration IC50 for DBP, DEHP and DINP is less than 500 ng / ml; and the generated antibodies have good generality and high sensitivity. The antigen or antibodies of the invention can be used for the establishment of enzyme-linked immunosorbent assay and colloidal gold test paper rapid detection methods, thus can be used for rapid detection of residues of phthalic acid ester plasticizers in food, and have broad application prospects.

The invention discloses a compound with an MEK (Mitogen-activated and Extracellular signal-regulated Kinase) inhibiting function as well as a preparation method and application of the compound. The structure of the compound disclosed by the invention is as shown in a formula (I). The preparation method of the compound disclosed by the invention comprises the steps of forming a coumarin ring by adopting a Pechmann reaction mainly and carrying out structural modification of different sites. In the binding experiment of the compound disclosed by the invention and MEK, the binding activity is up to 54.57nM; the anti-proliferation effect IC50 (half maximal inhibitory concentration) value on the melanoma cell A375 is up to 1.23 micrometers; the anti-proliferation effect IC50 value on the colon cancer cell HT-29 is up to 2.13 micrometers; and the activity is higher than that of a positive control U0126. The novel structure type coumarins compound with the MEK inhibiting function shows good MEK binding activity, MEK inhibiting activity, ERK (Extracellular signal Regulated Kinase) pathway inhibiting activity, anti-tumor effect and antiviral effect and has broad application value. The formula (I) is as shown in the specification.

The invention belongs to the medicinal field, and concretely relates to a use of a 8-nitrotryptanthrin compound as an IDO inhibitor. The 8-nitrotryptanthrin compound is a reversible IDO inhibitor, has an inhibition constant Ki of 0.054muM, has in-vitro and cell-based median effective inhibition concentrations IC50 of 0.103muM and 1.80*10<-5>muM respectively, and has an inhibition effectiveness obviously better than a present inhibitor 1-methyltryptophan (Ki of 34muM and IC50 of 340muM). 8-nitrotryptanthrin disclosed in the invention can effectively lower the abnormally-increasing IDO activity in a tumor animal model as the IDO inhibitor, and also has tumor treatment effects comprising tumor growth delaying, tumor volume reduction and in-vitro tumor cell killing. 8-nitrotryptanthrin disclosed in the invention has a wide application prospect, and can be used for treating serious diseases having the IDO mediated tryptophan metabolism approach pathology characteristics, such as cancers, the Alzheimer disease, tristimania, cataract and the like as the IDO inhibitor.

The invention discloses an enzyme linked immunosorbent assay method (ELISA) for detecting the total amount of diamond green and blank diamond green in measurement water sample and aquatic products, characterized in that: the modified compound of amido blank diamond green is synthesized and linked with the protein to produce immunogen and envelope antigen, and the polyclonal antibody of rabbit-anti colorless diamond green is obtained by immuning animal and the concentration range of ELISA standard curve is 0.1-100ng / mL, IC50 is 0.9-2.6ng / mL and the recovery of standard addition is 76.2-95%, the correlation coefficient of ELISA and HPLC is 0.975, n=7; because the cross-reaction rates of the prepared antibody and diamond green are respectively 95.25%, the ELISA can be used for measuring the total amount of the diamond green and blank diamond green without any oxidation step.

The present invention is used for evaluating the advantages of haliotis discus hannai. A multi-phase HPLC purification system is used to purify anti-inflammatory peptide from abalone (AAIP, abalone anti-inflammatory peptide). In tandem MS analysis, a fragmentation result shows that the amino acid sequence of the AAIP with nitrogen monoxide (NO) inhibitory activity (IC50=55.8[um]M) is Pro-Phe-Asn-Glu-Gly-Thr-Phe-Ala-Ser (1175.2Da). While the anti-inflammatory effect of RAW264.7 macrophages generated by the stimulus of the AAIP to lipopolysaccharides (LPS) is further studied, and the molecular mechanism is elaborated. The result shows that the AAIP peptide inhibits the nitrogen monoxide (NO) generation induced by the LPS through the expression of inducible nitric oxide synthase (iNOS) by a dose-dependent manner, and the gene transcription of pro-inflammatory cytokines is also obviously reduced, wherein the pro-inflammatory cytokines comprise such as interleukin (IL-1 beta), tumor necrosis factors (TNF-beta) and IL-6. In addition, the AAIP obviously inhibits phosphorylation of mitogen-activated protein kinases (MAPK), such as p-p38 and p-JNK. These results indicate that the AAIP inhibits inflammatory response induced by LPS by intercepting the MAPK pathway of the macrophages. Therefore, the AAIP can be applied to therapeutic drugs for inflammations treatment or healthcare food products.

Certain peptides which exhibit high selectivity for the kappa opioid receptor (KOR) versus the mu opioid receptor and little or no CYP3A4 inhibitory activity including tetrapeptides of four D-isomer amino acid residues having a C-terminus which is an N-oxide-substituted amide such, as H-D-Phe-D-Phe-D-Nle-D-Arg-NH-4-picolyl-N-oxide. A preferred compound, which has an affinity for the KOR at least 1,000 times its affinity for the mu opioid receptor and an IC50 for CYP3 A4 of greater than about 10 micromolar, is H-D-Phe-D-Phe-D-Nle-D-Arg-NH-4-picolyl-N-oxide

Ulcer is a serious oxidative stress induced disease with complex pathologic events including upregulation of H+K+ ATPase of parietal cell (PC) membrane, damage of mucin layer around PC, PC-DNA damage etc. The polysaccharide (GRPP) fraction and antioxidant extract (GRAOX) of ginger was examined for their ability to inhibit H+ K+ ATPase. Results indicated that the inhibition of H+ K+ ATPase activity which causes acidity in the lumen of the stomach, also exhibited better H+K+ ATPase inhibition (PPI) at the IC50 of 27.2 μg (GRPP) and 16.5 μg GAE (AOX) respectively than the known antiulcer proton pump blocker Lansoprazole (19.3 μg). Further the antioxidant activity in antioxidant extract (GRAOX) by various assay systems was examined such as Reducing power, Free radical scavenging (FRS), DNA and cytoprotection systems etc. GRAOX exhibited concentration dependent reducing power ability at 5-25 g equivalent of phenol. FRS activity with IC50 of 6.8 μg equivalent of phenol etc. At 0.3 μg level GRAOX offered >80% protection to DNA and against FeSO4-Asc'orbate induced oxidation. The major active phenolic components were identified as Gallic acid / Tannic acid (46%), and Cinnamic acid (51%).

The invention discloses an apple enzyme and a preparation method. The preparation method comprises the following steps: juicing apples, uniformly mixing apple juice with pure water, adding glucose which accounts for 1-5% of the total weight of the obtained raw material mixture into the raw material mixture that the volume percentage of the apple juice is 5-20%, adjusting the pH value to 5.0-7.0, sterilizing, preparing mixed bacteria from the obtained raw material liquid and a pichia pastoris bacterial liquid and a lactobacillus plantarum bacterial liquid in a volume ratio of 1:(1-5), controlling the temperature to 25-35 DEG C, performing primary fermentation for 12-48 hours, controlling the temperature of the obtained primary fermentation solution to 25-35 DEG C by using an acetic bacterial liquid, performing secondary fermentation for 12-36 hours, and sterilizing, thereby obtaining a finished product of the apple enzyme. Compared with a finished product of natural fermentation, the finished product of the apple enzyme disclosed by the invention has a remarkable antibacterial effect on propionibacterium acnes, and the removal rate IC50 of the apple enzyme on DPPH is 0.43%.

The invention discloses a method for comprehensive development and utilization of fruits and vegetables. The method uses fruits and vegetables as raw materials and comprises separating fruit and vegetable juices from fruit and vegetable residues by juicing, carrying out fermentation on the fruit and vegetable juices through a compound liquid of three edible and medical fungi such as needle mushroom, black fungus and grifola frondosa to obtain an antioxidant activity fermented fruit and vegetable juice, wherein in the DPPH free radical scavenging system, IC50 is 40 to 100 microliters per milliliter, and carrying out fermentation on the fruit and vegetable residues through lactic acid bacterium compound solids such as lactobacillus plantarum, lactobacillus acidophilus and lactobacillus rhamnosus to obtain fruit and vegetable ferment having a blood sugar reduction function. An in-vitro blood sugar reduction experiment result shows that the fruit and vegetable ferment has an alpha-glucosidase inhibition rate of 40-55%. The method provides a novel approach for comprehensive development and utilization of fruits and vegetables, can produce the fruit and vegetable juice having antioxidant activity and can produce the fruit and vegetable ferment having a blood sugar reduction function.

The invention relates to a pseudomonas aeruginosa bacterial strain (serial number being CB-4) and a crude toxin preparation method thereof, which belong to the field of applying microorganisms to agricultural plant protection. The strain crude toxin has a strong inhibition effect for weeds including crab grass, amaranthus retroflexus, barnyard grass, green bristlegrass, rhodes grass and the like, and in particular respectively has IC50 reaching 299.25mg.L<-1> and 210.11mg.L<-> to seed germination root stalk length, and has the characteristics of being safe and environment-friendly, harmless to crops and the like.

The invention discloses a synthesis method of a specific salbutamol artificial antigen, belonging to the technical field of biological chemical engineering. The synthesis method disclosed by the invention comprises the following steps of: activating salbutamol through a formaldehyde solution, and connecting 6-aminocaproic acid in the ortho-position of a phenolic hydroxyl group to obtain a salbutamol hapten; and coupling a carboxyl group on the salbutamol hapten with an amino group on a carrier protein to obtain the salbutamol artificial antigen. The synthesis method disclosed by the invention can make up for the insufficiencies and defects of the existing salbutamol antigen synthesis technologies, the salbutamol artificial antigen with high specificity is obtained, the specificity of a produced antibody is high, and the sensitivity is high; and experimental results show that the antiserum titre of an animal immunized by using the salbutamol artificial antigen disclosed by the invention can achieve 80000, the detection limit is 0.5ng / mL, and the half-inhibitory concentration IC50 is 5ng / mL. The antigen or the antibody disclosed by the invention can be used for establishing an enzyme-linked immunosorbent analytical method and a colloidal gold test strip rapid assay method so as to rapidly detect the residues of the salbutamol in a food and further realize broad application prospects.

The invention relates to a method for continuously preparing silkworm chrysalis protein antioxidant peptides by using an enzyme membrane reactor. The method comprises the following steps of: firstly, preparing silkworm chrysalis protein into a suspension by using distilled water; secondly, homogenizing the suspension by using a homogenizer, and then adding the suspension together with incision enzyme into the enzyme membrane reactor for continuously preparing antioxidant peptides; thirdly, penetrating the enzymolysis product solution through an ultrafiltration membrane to ensure that the silkworm chrysalis protein which is not fully reacted and the incision enzyme are captured and then reflow into a reaction tank for continuous reaction; and finally, collecting the enzymolysis product penetrating fluid penetrating through the ultrafiltration membrane, and concentrating and then carrying out spray drying to obtain a powdered silkworm chrysalis protein antioxidant peptide product. The silkworm chrysalis protein antioxidant peptide prepared by the method is good in activity, the IC50 for clearing DPPH (1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl) free radical is 0.9-1.6mg / mL, the IC50 of the capacity of clearing hydroxy radical is 0.2-0.6mg / mL, and the IC50 of the capacity of chelating Fe<2+> is 1.4-2.7mg / mL.

The invention discloses a method for measuring nano-gold mimetic peroxidase based urease and an inhibitor thereof. The method is characterized by comprising the following steps: coupling a urease catalysis urea reaction system with a nano-gold mimetic peroxidase-hydrogen peroxide-3,3',5,5'-tetramethylbenzidine dihydrochloride catalytic chromogenic reaction system, and combining with the changes of solution colors and ultraviolet absorption spectrum characteristics to directly measure the content of urease and the inhibitor thereof, wherein in a range of 1.8-90U / L, A450 and the urease concentration have a linear relationship, and the limit of detection is 1.8U / L; and by virtue of software fitting, IC50 of obtained acetohydroxamic acid of the urease inhibitor is 0.05mmoL / L. The method disclosed by the invention is simple and convenient to operate and high in sensitivity, and can be applied to the measurement of the content of urease and the inhibitor thereof in environment and life science systems as an analysis method.

The present invention discloses a preparation method for increasing antioxidation activity of flammulina velutipes polysaccharide, wherein an enzymolysis control method is adopted to regulate and control molecular weight distribution of flammulina velutipes polysaccharide, flammulina velutipes is sequentially subjected to crushing, impurity removing, hot water extraction, protein removing, concentration and ethanol fractional precipitation separation to obtain the flammulina velutipes polysaccharide, and an enzyme method is adopted to carry out degradation control so as to reduce a molecular weight of the flammulina velutipes polysaccharide to 19000-30000 Da. The flammulina velutipes polysaccharide prepared by using the method is suitable for industrial production, wherein a polysaccharide extraction rate is increased by 2.5-3.5 times compared with a polysaccharide extraction rate of the traditional water extraction method, significant antioxidation activity is provided, IC50 of the flammulina velutipes polysaccharide in a DPPH.radical system is 1.69-2.12 mg / mL, and an ORAC value can be up to 923 mumol Trolox / g.

The invention relates to a rhodamine B ELISA detection method, which belongs to the technical field of ELISA. The invention discloses a rhodamine B ELISA detection method, which comprises the following steps: using synthesized rhodamine B immunogen for immunizing healthy New Zealand rabbits to obtain polyclonal antibodies; and using rhodamine B as standard products; using rhodamine B hapten and OVA coupling materials as coating antigen to establish a rhodamine B indirect ELISA method. The invention provides the fast and efficient detection method for the residual detection on the rhodamine B. Because the polyclonal antibodies are adopted, the cost is low, and in addition, the stability and the repetitiveness are good. The detectability (IC90) is 0.028 ng / mL, the half inhibitory amount (IC50) is 1.3 ng / mL, and the detection range (IC20 to IC80) is between 0.07 and 17.5 ng / mL. Because of high specificity and high compatibility of the immune reaction, high selectivity and high sensitivity can be obtained when the ELISA is used for detecting the rhodamine B.

The invention discloses antitumor platinum drug mineralization protein nanoparticles and a preparation method and application thereof. The antitumor platinum drug mineralization protein nanoparticlesand the preparation method thereof have the advantages that the platinum drug mineralization protein nanoparticles are successfully prepared through the two-step preparation method for the first time,the prepared nanoparticles can be uniformly dispersed in an aqueous solution, the nanoparticles has good cytotoxicity, and the IC50 of the nanoparticles to human non-small-cell lung cancer cells A549is 6.9 microgram / ml; the platinum drug mineralization protein nanoparticles comprising a platinum drug and protein are simple in preparation process, uniform in size, controllable in particle size, good in biocompatibility, good in water solubility, long in blood circulation time, high in tumor targeting performance and capable of laying a foundation for efficient tumor treatment or drug-resistant tumor treatment.

The invention discloses a method for preparing an antioxidative peptide from skipjack processing by-products. The method comprises the following steps of mixing head sections and tail sections as by-products rich in proteins and a stewing liquor, crushing the mixture, carrying out synchronous three-phase separation to obtain a protein source which can be subjected to direct enzymolysis, adding a flavourzyme into the protein source, carrying out enzymolysis at a pH value of 6.5 at a temperature of 50 DEG C for 4h, purifying the enzymatic hydrolysate by ultrafiltration, activated carbon adsorption and desalination, and carrying out spray drying to obtain peptide powder. Various antioxidant activities of the peptide powder are detected. The peptide powder has good DPPH free radical, superoxide anion free radical and hydroxyl radical removal rates, has IC50 of 3.55mg / ml, 1.39mg / ml and 0.61mg / ml and is an excellent anti-oxidant. The peptide powder is flavescent, has no bitter taste, has a light fishiness taste, has molecular weight (greater than 90.1%) less than or equal to 10000Da, has polypeptide total protein content of 88.6% by dry weight and has ash content of 7.2%.