83 results about "Caco-2" patented technology

The invention discloses a caco-2 cell model for CRISPR / CAS9-mediated drug transporter targeted knockout and a method thereof. The method is used for non-diagnostic or therapeutic purposes and comprises the following steps: designing sgRNA with target specificity for P-gp, BCRP and MRP2 transporters and constructing a sgRNA expression vector, wherein a sequence of the designed sgRNA is shown as SEQID NO. 1-6 in a sequence table; respectively designing P-gp, BCRP and MRP gene single knockout and pairwise combination double knockout by utilizing CRISPR / CAS9, co-transfecting a caco-2 cell with anhCas9 plasmid and performing monoclonal expansion culture to obtain the caco-2 cell model for transporter gene targeting. The caco-2 cell model obtained by the method disclosed by the invention has the beneficial effects that the mutual interference among different transporters is effectively eliminated, and a more specific and more sensitive cell model is provided for drug transport research.

The invention discloses the nanoparticles composed of chitosan / poly-γ-glutamic acid characterized with a positive surface charge and their enhanced permeability through Caco-2 cells for paracellular drug delivery.

The invention relates to effective fractions of cicer arietinum linn bean sprouts and a preparation method and application thereof. The effective fractions of the cicer arietinum linn bean sprouts, namely total saponin and total isoflavone are obtained by a plant chemical extraction separation method and a macroporous resin enrichment process. A bioactivity screening result indicates that the effective fractions of the cicer arietinum linn bean sprouts, namely the total saponin and the total isoflavone can suppress the growth of human adenocarcinoma colon strain Caco-2 cells by inducing cell apoptosis and are used for effectively inducing human adenocarcinoma colon cell apoptosis. A new thought for the research and development of the application of the effective fractions to the preparation of a medicament or a health-care product for treating colon cancer is provided.

Targeting metabolic enzymes in human cancer Abstract Lung cancer is a devastating disease and a major therapeutic burden with poor prognosis. The functional heterogeneity of lung cancer (different tumor formation ability in bulk of tumor) is highly related with clinical chemoresistance and relapse. Here we find that, glycine dehydrogenase (GLDC), one of the metabolic enzyme involved in glycine metabolism, is overexpressed in various subtypes of human lung cancer and possibly several other types of cancers. GLDC was found to be highly expressed in tumor-initiating subpopulation of human lung cancer cells compared with non-tumorigenic subpopulation. By array studies we showed that normal lung cells express low levels of GLDC compared to xenograft and primary tumor. Functional studies showed that RNAi inhibition of GLDC inhibits significantly the clonal growth of tumor-initiating cells in vitro and tumor formation in immunodeficient mice. Overexpression of GLDC in non-tumorigenic subpopulation convert the cells to become tumorigenic. Furthermore, over-expression of GLDC in NIH / 3T3 cells and human primary lung fibroblasts can transform these cells, displaying anchorage-independent growth in soft agar and tumor-forming in mice. Not only is GLDC is expressed human lung cancer, it is also up-regulated in other types of cancer, such as colon cancer. RNAi knockdown of GLDC in colon cancer cell line, CACO-2 cells, can also inhibit the tumor formation in mice. Thus GLDC maybe a new metabolic target for treatment of lung cancer, and other cancers.

The invention belongs to the field of medical biotechnology, and relates to two series major vault protein of glucagon-like peptide-2 (GLP-2) of people and a preparation method of the two series major vault protein. A second place of an amino acid fragment of the glucagon-like peptide-2 of people is mutated, two segments are connected in series through connecting peptide, a dimmer structure of the GLP-2 is simulated, encoding gene of the two series major vault protein of the GLP-2 is obtained by a synthetic method, the encoding gene is cloned to a pET22b(+) pronucleus so as to express a vector, Escherichia coli BL21 is transformed, the encoding gene is expressed in the Escherichia coli efficiently, the purified reconstructed purpose protein is obtained through salt fractionation of ammonium sulfate in fractional and chromatography of anion exchange columns. The reconstructed purpose protein can stimulate a Caco-2 cell, multiplication of the Caco-2 cell can be promoted notably, the two series major vault protein of GLP-2 of people has natural biological activities, and therefore a foundation is established for further study and extensive use of the GLP-2 of people.

The invention discloses a yeast hollow beta-glucan shell wrapped GMP-BSA protein targeting macrophage delivery system. The system allows BSA to be coated in a yeast shell through using a compact layer formed through the electrostatic adsorption effect of chitosan (CS), tripolyphosphate (TPP), sodium alginate and BSA. GMP-BSA particles prepared in the invention have very good protein release behavior in in-vitro experiments, and can avoid protein loss. The particles highly specifically target the macrophages, such as Raw 264.7 cells, primary BMDM cells (primary bone marrow macrophages) and peritoneal macrophages (PEMs), and are not devoured by NIH3T3, AD293, HeLa, Caco-2 and other non-macrophage tumor cells, and neutral granulocytes in blood. The macrophages play a great role in the pathogenesis of various diseases, and are very important potential target spots. The macrophages are key antigen transfer cells, and are very important in vaccine design. The system can highly selectively deliver various proteins to the macrophages in a targeting manner, and also has very wide application prospect in the field of targeting drug administration and the field of macrophage vaccine design.

The invention discloses a method for quantitatively evaluating the antioxidant activity of an antioxidant based on a Caco-2 cell model, relating to an antioxidant. The method for cytologically and quantitatively evaluating the antioxidant activity of an antioxidant based on a Caco-2 single-layer cell model is established by using the Caco-2 cell model and optimizing the inoculation concentration and culture time of cells as well as the incubation conditions of a fluorescent probe (2',7'-dichlorodihydrofluorescein diacetate, DCFH-DA for short), a free radical initiator (2,2'-azobis(2-methylpropionamidine)dihydrochloride, AAPH for short), a pure antioxidant and Caco-2 cells. The method has the advantages of high biological relevance, easiness and high speed, and an effective analysis and evaluation tool is provided for the research of biological antioxidants.

The invention discloses a construction method for a human intestinal tract epithelial cell model, and relates to the technical field of cell model construction. The human intestinal tract epithelial cell model is characterized in that a Caco-2 cell is taken as a model cell, and a DMEM (dulbecco's modified eagle medium) containing 20%FBS (fetal calf serum) is used for carrying out 3D culture on theCaco-2 cell in a micro-fluidic chip so as to construct the in vitro model of the human intestinal tract epithelial cell. On one hand, a human cell is adopted for culture, the problem that an experiment result is in accurate since an animal cell is adopted for culture is avoided, and on the other hand, the 3D cell culture enables intestinal tract cells to own a mechanical microenvironment which ismore similar to the human physiology, so that intestinal tract cells can be accelerated to differentiate. Therefore, the human intestinal tract epithelial cell model constructed by the construction method disclosed by the invention can more clearly and accurately reflect the characteristics of small intestine epithelial barriers.

The invention discloses a food allergen dynamic digestion model and an in-vitro simulation evaluation method. The method mainly comprises the steps of establishing an allergen dynamic digestion modeland an in-vitro simulation evaluation method, simulating a biochemical reaction condition of the stomach and duodenum of a human body through a digestive system simulation device, providing a method for constructing an allergen in-vitro simulated dynamic digestion experiment in the simulated evaluation method, constructing a Caco-2 (human cloned colonic adenocarcinoma cells) small intestine epithelial cell model to simulate small intestine absorption and transport effects, and constructing a KU812 (human peripheral blood basophilic leukemia cells) cell model to evaluate the ability of mast cell degranulation induced by food allergen digestion and transport products. The method of the invention is suitable for evaluating the digestion and sensitization of a purified food allergen or an allergen extracting solution containing a complex food matrix.

The invention relates to a calcium chelating peptide as well as a preparation method and an application thereof. An amino acid sequence of the calcium chelating peptide is FDHIVY. The calcium chelating peptide provided by the invention is safe and non-toxic, and has better physicochemical activity compared with a traditional calcium supplement, and the calcium chelating capability of the calcium chelating peptide reaches 35.73 mg / g; the collagen peptide can be supplemented while absorption and bioavailability of calcium in a human body are improved, the absorption and transport of calcium in Caco-2 small intestinal epithelial cells can be promoted, and the calcium chelating peptide can be used as a raw material for drug development and biological calcium supplementation agents. According to the preparation method provided by the invention, the calcium chelating peptide is successfully obtained, fish product resources can be fully utilized, the operation is simple, and a new way is provided for high-valued utilization of tilapia bones.

The invention provides a bio-safety evaluation method of nano zinc oxide based on Caco-2 cells. The method comprises the following steps: conventionally culturing cells, passaging the cells, freezing the cells, thawing the cells, contaminating the cells, performing morphological observation and detecting the cell activity. The method has certain theoretical meaning on discussion on intestinal toxicity of nano zinc oxide; a reference is provided for establishment of food nano zinc oxide limit standards, a reliable scientific basis is provided for further study on the safety problem of food nano zinc oxide, a reference is provided for safety evaluation of other nanomaterials, and theoretical support is provided for safety use of nano zinc oxide.

The invention provides application of a preclinical pharmacokinetic key technology and research system in cefoperazone sodium and sulbactam sodium, comprising the steps of (1) after the cefoperazone sodium and sulbactam sodium with a certain concentration is administrated to an experimental animal for a certain time, collecting one or more biological samples in blood, urine and faeces; (2) treating the biological samples obtained in step (1) by adopting liquid-liquid extraction, albumen precipitation, and solid phase extraction technologies to obtain corresponding solutions; (3) analyzing the prepared solutions in step (2) by adopting an LC-MS(liquid chromatography-mass spectrography) and an LC-MS / MS((liquid chromatography-tandem mass spectrometry). According to the application provided by the invention, the Caco-2 cell can also be adopted to detect the membrane permeability of the medicine and cell absorptive capacity; or the medicine with a certain concentration treats primary culture cerebral microvascular endothelial cells to detect the blood-brain barrier permeability of the medicine; or by adopting a whole animal, S9, human intestinal microsome and monoclonal purified enzyme, in vitro metabolism stability of the medicine is detected and a metabolite is identified.

The invention provides application of vacuolar protein sorting 4A (VPS4A) to preparing medicines for preventing or treating enterovirus type 71 infection, relates to the technical field of biomedicine, and provides novel enterovirus type 71 infection resisting targets and application. The application includes that expression of host proteins of target cells which are human colon cancer cells (Caco-2) is down-regulated by the aid of an RNA (ribonucleic acid) interference technology, so that host factors capable of effectively inhibiting EV71 (enterovirus type 71)-infected human colon cancer cells (Caco-2) can be searched, and the purpose of effectively eliminating EV71 infection from sources (intestinal tracts) can be achieved. The application has the advantages that as discovered via experiments, important effects can be realized by the vacuolar protein sorting 4homolog A (VPS4A) for the EV71-infected Caco-2, expression of the VPS4A is down-regulated, and accordingly EV71 infection can be obviously inhibited.