59 results about "Small intestinal epithelium" patented technology

The invention relates to the technical field of microorganisms, and particularly provides Lactobacillus reuteri and application thereof. The Lactobacillus reuteri provided by the invention is a canine-derived Lactobacillus reuteri strain which is separated from hindgut content of a healthy adult blue fox for the first time by an inventor, and the preservation number of the Lactobacillus reuteri isCCTCC NO: M 2018943. The strain has good growth performance, is high in tolerance to gastric juice and intestinal juice and good in adhesion to small intestine epithelial cells, can promote proliferation of dog intestinal tract beneficial bacteria, can inhibit growth of harmful bacteria, can enhance the immune function of pet dog organisms, has better effect than that of similar products in the market, can serve as an alternative strain of probiotic preparations, and is quite wide in application prospect.

The invention provides an establishment of humanized three-dimensional intestinal mucosa model and an application thereof. The model improves a traditional Caco-2 monolayer cell model mainly based on normal physiological structure characteristics of small intestinal epithelium, wherein on basis of adding extracellular matrix, the Caco-2 cells are co-cultured with mucus-secreting cells, interstitial cells and immune cells for a period of time to form a three-dimensional intestinal mucosa model being similar with structure and function of the small intestinal epithelium in vivo. By using the model, a membrane permeating capability at small intestine epithelium as well as a metabolism degree of oral drug molecules can be predicted. The model is mainly used for researching transmembrane transportation and absorption, and metabolism process of medicament molecules at the small intestine in vitro.

The invention provides a rabbit intestinal epithelial cell line. The preparation method comprises the following steps: the ileum part of a newborn baby rabbit is taken to be mechanically cut to pieces, collagenase IV is used to digest the cut ileum part to obtain primary cultured rabbit intestinal epithelial cells, the phase contrast digestion method is combined with the phase contrast adherent method to perform primary purification on the rabbit intestinal epithelial cells; then the monoclonal method is adopted to further purify the cells, and the obtained cell line grows well and stably; the shape of the cells is regular, the cells show a flagstone appearance, and the keratin 8 identification result of the cell line is positive; and a cell line is screened out through subculture and thesubculture is kept stable. The rabbit intestinal epithelial cell line comes from the ileum part of the newborn Japanese big-ear rabbit, the preservation number is CGMCC ((China General Microbiological Culture Collection Center) No.4784; and the cell line can be used in the absorption pathway of nutrient substances in the intestinal tract, have the function of resisting nutrient substances and be used in the researches of the microecological nutrition, the signal transduction between cells, the proliferation and differentiation mechanisms of various cells in a crypt-villus system, etc.

The invention belongs to the technical field of cell isolation and cultivation. In order to solve the problems that the chicken small intestinal epithelial cells are difficult to isolate and culture in vitro at present, the existing isolation process is troublesome and time-consuming, the obtained cells are low in purity and the like, the invention provides isolation and primary culture methods of the chicken small intestinal epithelial cells. The methods comprise the steps of taking chick embryo small intestines, cleaning the small intestines to remove mesenteries, digesting small intestine tissues by protease to obtain the chicken small intestinal epithelial cells, performing adherent difference culture on the cells, removing adherent parenchyma cells, collecting intestinal epithelial cells, inoculating by taking the inoculum density being about 6.5X105 / mL for culturing, the chicken small intestinal epithelial cells obtained by culturing is maximum in quantity, low in death rate, and best in growing status. The primary culture is performed by adopting a culture solution in which chicken serum is added, and the growing status of the obtained cells is better. The chicken small intestinal epithelial cells which are good in growing status, normal in shape, high in activity are successfully cultured, the primary and isolation culture methods of the chicken small intestinal epithelial cells are successfully established, and the technical support is provided for follow-up tests.

The invention discloses corn prolamin anti-inflammatory polypeptide and a preparation method thereof, and belongs to the technical fields of deep processing of agricultural products and preparation offunctional food. The preparation method comprises the following steps: performing ultrasonic pretreatment on corn prolamin, performing enzymolysis by using protease to prepare corn anti-inflammatorypolypeptide, tracking the activity of the corn anti-inflammatory polypeptide by simulating gastrointestinal digestion, simulating intestinal epithelial cell absorption through Caco-2 cells and representing the corn pure peptide with high anti-inflammatory activity capable of resisting Caco-2 cell decomposition through gastrointestinal digestion and being absorbed by the inner wall of the small intestine simulated by the Caco-2 cells. The invention identifies the corn prolamin functional polypeptide with high anti-inflammatory activity for the first time, wherein the corn prolamin functional polypeptide can resist Caco-2 cell decomposition through gastrointestinal digestion and can be absorbed by the inner wall of the small intestine simulated by the Caco-2 cells. Through combination of thesimulated digestion system and the Caco-2 simulated intestine inner wall absorbing system, the change of the activity of the corn anti-inflammatory polypeptide is tracked; and the absorption rate ofthe corn polypeptide digestion peptide in the Caco-2 simulated intestine inner wall absorbing system is researched.

The invention discloses a piglet small intestine epithelial cell classification and separation method, which includes the following steps: (1) a small intestine section which is 60cm to 90cm long is adopted, 100mL to 150mL of 37 DEG C preheated and oxygenated phosphate buffer is injected into the enteric cavity, both ends are clipped by hemostatic forceps, the small intestine section is then incubated in a 37 DEG C water bath oscillator for 30 minutes, and the speed of oscillation is 70 rotations per minute; (2) the phosphate buffer is removed, 100mL to 150mL of cell separation solution is injected, the small intestine section continues to be incubated in the 37 DEG C water bath oscillator for 2.5 hours, the speed of oscillation is 70 rotations per minute, and the cell separation solution is collected respectively at different time points and new cell separation solution is injected; (3) the collected cell separation solution is centrifuged at 400g under 4 DEG C for ten minutes, the obtained precipitate is intestinal epithelial cells, and the cells which are collected according to the sequence of incubation times are cells of different parts from the villus tip to the crypt bottom in sequence along a recess-villus axis. The cells separated by the invention provide a novel technical method for an in-vivo study on the small intestine epithelial cell differentiation mechanism, the absorption and metabolism of nutrition and the development of drugs for related diseases.

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 provides a PGC promoter of porcine small intestinal epithelial cell expression and application thereof. The PGC promoter has the nucleotide sequence shown in SEQ ID No.1. A PGC reliablepromoter area is predicted through an online website; a primer is synthesized through a chemical synthesizing method; a PGC promoter sequence is inserted in a pGL3-Basic carrier through restriction enzyme cutting sites MluI and NheI to obtain a PGC-dual-luciferase recombinant plasmid; the plasmid is transfected into escherichia coli to be amplified, the plasmid is extracted, and sequencing and verifying are conducted; the plasmid is transfected into a porcine small intestinal epithelial cell, it is found that the porcine PGC promoter can promote the dual-luciferase gene expression in the porcine small intestinal epithelial cell, it is shown that the promoter can regulate and control the expression of a downstream gene in the porcine small intestinal epithelial cell, a stable and efficientsmall intestinal transfection regulation and control element can be easily established, and the PGC promoter has application value in the research on the transgenic animal preparation and the small intestinal disease gene therapy.

The invention relates to a sulfide quinone oxidoreductase intestinal tract directed expression vector and a cell line thereof, belonging to the field of biotechnology. The SQR (sulfur quinine oxidoreductase) gene is inserted into a eukaryotic expression vector pcDNA (deoxyribonucleic acid) 3.1 (1), so that a slow virus recombinant vector Lanti4-blockit-IFABP (intestinal fatty acid binding protein)-SQR-GFP (green fluorescent protein) can be built, and the primarily cultured pig intestinal epithelial cell is infected by the slow virus recombinant vector, so that the eukaryotic cell line of specific expression SQR-GFP protein can be built. In order to further prepare the somatic cell nuclear transfer clone trans-SQR gene, necessary transgenic biological materials and key technical system support can be provided for cultivating new transgenic pig species reducing the pollutant discharge.

The invention discloses an in-vitro culture method for small intestinal epithelial cells of a calf. A small intestine of the calf is taken, an intestinal canal is guided into an intestinal canal dilator so as to be dilated, manure in the small intestine is washed with a PBS fluid until a liquid flowing out is clear, then one end of the intestinal canal is sealed with a pair of haemostatic forceps, the other end of the intestinal canal is sealed after the intestinal canal is filled with a protease digestive fluid, the intestinal canal is put onto a shaker to be digested at the temperature of 37 DEG C for 1-1.5 h, a liquid is collected and washed twice in a centrifugal manner, and finally, a culture solution is added for inoculated culturing; cells grow against the wall of the intestinal canal after 12-24 h, 90% of cells are attached to the wall of the intestinal canal after 3-4 days, and the cells are the small intestinal epithelial cells. The protease digestive fluid is directly injected into the enteric cavity and is not contacted with the outer wall of the small intestine, accordingly, fibroblasts and muscle cells on the outer wall of the intestine cannot be digested, and the purity of the epithelial cells is guaranteed to the largest extent.

The invention relates to a sulfide quinone oxidoreductase intestinal tract directed expression vector and a cell line thereof, belonging to the field of biotechnology. The SQR (sulfur quinine oxidoreductase) gene is inserted into a eukaryotic expression vector pcDNA (deoxyribonucleic acid) 3.1 (1), so that a slow virus recombinant vector Lanti4-blockit-IFABP (intestinal fatty acid binding protein)-SQR-GFP (green fluorescent protein) can be built, and the primarily cultured pig intestinal epithelial cell is infected by the slow virus recombinant vector, so that the eukaryotic cell line of specific expression SQR-GFP protein can be built. In order to further prepare the somatic cell nuclear transfer clone trans-SQR gene, necessary transgenic biological materials and key technical system support can be provided for cultivating new transgenic pig species reducing the pollutant discharge.

The invention provides a recombinant staphylococcus aureus enterotoxin G oral preparation which has an amino acid sequence of SEQ ID NO.1 and also comprises excipients in medicine or carriers allowed by the preparation. By the experiment on the Caco-2 monolayer cell trans-membrane transport, the preparation of the invention proves that the proteins can enter the general blood circulation from intestinal epithelial cells in the form of complete molecules and keep promoting the splenic lymphocyte proliferation and preventing the superantigen activity for the growth of tumor cells, and can be applied to the preparation of drugs for curing the malignant tumor and other serious complications.

The invention discloses a porcine epithelial cell oxidative stress model, an establishment method therefor and application of the porcine epithelial cell oxidative stress model. The establishment method for the porcine epithelial cell oxidative stress model comprises the following steps: inducing porcine epithelial cells by AAPH to continuously generate stable endogenous free radicals, detecting the oxidative damage degree of the cells, and determining a stress model of oxidative damage. According to the porcine epithelial cell oxidative stress model, the optimal concentration for constructingthe porcine small intestinal epithelial cell oxidative stress model induced by the AAPH is determined by a series of tests, and the oxidative damage is evaluated by the established oxidative stress model, so that the convenient and stable intestinal epithelial cell model is provided for further research of the influence of the oxidative stress on animal intestinal health, and the model has important significance for maintaining human health and reducing direct losses and indirect losses caused by oxidative stress in animal husbandry production.

The invention provides an establishment method of a heavy metal oral bioaccessibility detection system. The establishment method comprises the following steps: 1) respectively establishing a human body external gastrointestinal digestion model and a human small intestine epithelial cell 3D model; 2) respectively applying substances containing different concentrations of heavy metals to the human body external gastrointestinal digestion model to simulate the gastrointestinal digestion process; 3) respectively applying the digestive juice containing the heavy metals to the human small intestineepithelial cell 3D model, and performing culturing for a preset time to simulate a small intestine absorption process; and 4) tracking changes of heavy metal concentrations and valence states in the human body external gastrointestinal digestion process and the small intestine epithelial cell absorption process, calculating oral biological accessibility of heavy metals, and establishing a method for predicting exposure risks based on the biological accessibility. The invention belongs to the technical field of food safety detection and evaluation, is suitable for detecting oral biological accessibility of heavy metals, and is also suitable for potential toxicity effect detection and exposure risk evaluation.

The invention discloses a method for constructing a conditional inducible expression AsCpf1 lentiviral vector. The invention also discloses a Tet-on-3G-pLVX-AsCpf1 lentivirus obtained by packaging thelentiviral vector and an application of the lentivirus to pig small intestine epithelial cell line construction. Pig small intestine epithelial cells are a cell material that is widely used in the study of intestinal nutrients and immunity. The present invention utilizes an AsCpf1-expressed lentivirus system regulated by a Tet-on-3G system to screen out an inducible-expressed stable-transfectionpig small intestine epithelial cell line. When Dox is added for induction, cells start a large amount of AsCpf1 expression within 24 hours, and when no Dox is added for induction, AsCpf1 expression disappears; and AsCpf1 decreases significantly after the Dox adding is stopped for 12h, AsCpf1 expression level is extremely low after the Dox adding is stopped for 24h and 48h, and no expression can beobserved after 72h.

The invention provides a new application of tauroursodeoxycholic acid (TUDCA) or pharmaceutically acceptable salts thereof in preparation of drugs for prevention or treatment of neonatal necrotic enterocolitis. TUDCA plays a good protective role on endoplasmic reticulum stress caused by neonatal Necrotic Enterocolitis (NEC), CHOP and BiP are significantly reduced under the action of TUDCA, TUDCA intervention can maintain the weight of NEC model newborn mice, reduce intestinal epithelial cell apoptosis, and improve the survival rate of NEC model newborn mice. Through cell and apoptosis level experiments, TUDCA is confirmed to play a protective role on NEC by inhibiting endoplasmic reticulum stress and reducing the cell apoptosis proportion in NEC, and lays a theoretical foundation for prevention and treatment of neonatal necrotic enterocolitis by TUDCA.

The invention discloses a liquid calcium soft capsule and a preparation method thereof. The liquid calcium soft capsule comprises a capsule shell and contents in a capsule body, the capsule skin comprises 30%-50% of gelatin, 10%-30% of glycerol, 30%-50% of purified water and 1%-4% of a coloring agent; the content in the capsule body comprises 30%-50% of calcium citrate, 1%-5% of milk mineral salt, 0.1%-3% of casein phosphopeptides, 0.01%-2% of vitamin D3 and 40%-60% of soybean oil. Calcium citrate is used as a main calcium source, and milk mineral salt is used as a secondary calcium source. Calcium citrate is organic acid calcium, the absorption rate of the calcium citrate is higher than that of inorganic calcium, the content of citric acid in urine can be increased, the urine is acidified, and formation of calculi is inhibited. Milk mineral salt is separated from milk, irritation to intestines and stomach is small, and the calcium supplement effect cannot be reversed. Casein phosphopeptides are added and combined with Ca < 2 + > to form a soluble complex, so that Ca < 2 + > is prevented from being precipitated in neutral or weakly alkaline small intestines, and absorption and utilization of small intestine epithelial cells to Ca < 2 + > are promoted. Vitamin D3 is a hormone precursor acting on calcium and phosphorus metabolism, and can improve the absorption of the body to calcium and phosphorus.

The invention discloses a pig intestinal epithelial cell promoter SIECP and application thereof. the disclosed pig intestinal epithelial cell promoter SIECP is a DNA molecule shown in a sequence 1 ina sequence table. The SIECP can activate the target gene expression in intestinal epithelial cells, and can be used for cultivating transgenic animals, theoretical basis is provided for constructing stable and efficient intestine transcription regulatory elements, reference value is achieved for the production of environment-friendly transgenic animals and the gene therapy research of intestine diseases, and wide application prospects in the field of life science and intestine disease treatment are achieved.

The invention discloses a zein ACE inhibitory peptide and its application as a health food, and belongs to the technical field of intensive processing of agricultural products and preparation of functional foods. In the present invention, ultrasonic pretreatment of zein is firstly performed, followed by protease enzymatic hydrolysis to prepare zein ACE inhibitory active peptide, and then the ACE inhibitory activity of zein active peptide is tracked by simulating gastrointestinal tract digestion, and then Caco‑2 Following absorption by cells mimicking the intestinal epithelium, pure zein peptides with high ACE inhibitory activity were characterized after gastrointestinal digestion and absorbed by Caco‑2 cells mimicking the lining of the small intestine. The present invention first studies the absorption rate of zein polypeptide digestion products in the Caco-2 simulated small intestinal wall absorption system; for the first time, it identifies two kinds of zein peptides that have been digested by the gastrointestinal tract and absorbed by Caco-2 cells to simulate the small intestinal wall. ACE-inhibiting active zein functional polypeptides.

The invention discloses a fucoidin-containing composite preparation and application thereof, and belongs to the field of health care products. The fucoidin composite preparation consists of the following components in parts by weight: 9-80 parts of fucoidin, 1-30 parts of magnolol, 1-30 parts of atractylodes macrocephala koidz oil and 1-20 parts of chlorogenic acid. A zebrafish macrophages PM 2.5phagocytosis is used as a research object, through the research, it discovers that the fucoidin-containing composite preparation can remarkably increase the number of macrophages, the phagocytic ability of the macrophages is promoted, movement of the macrophages to intestinal tracts is promoted, intestinal epithelial cells discharge fine particulate matters, thus, the fine particulate matters aredischarged out of the body through the intestinal tracts, and thus, the purpose of reducing ultra-micro particulate matters in the body is achieved.

The invention relates to an immortalized tree shrew small intestinal epithelial cell line and its construction method and application, belonging to the field of cell technology. The present invention activates the telomerase activity of tree shrew intestinal epithelial cells by using lentivirus-mediated hTERT gene transfection to obtain the cell line, and can rapidly and effectively establish immortalized tree shrew intestinal epithelial cells, and the established immortalized Tree shrew intestinal epithelial cells have successfully overcome replicative senescence and acquired immortality, and the immortalized tree shrew intestinal epithelial cells have maintained a phenotype very similar to that of primary tree shrew intestinal epithelial cells, maintaining cell contact inhibition and cell proliferation. Density is limited, and CPE can appear after infection with reovirus, so it can be used for research on virus infection and the like.

The invention discloses an additive for improving the jejunum anti-oxidation capacity of piglets with intrauterine growth retardation, and a preparation method and application of the additive, and belongs to the technical field of feeds. The additive is prepared from the raw materials in parts by weight: 10-15 parts of edible oil, 10-20 parts of an antioxidant, 5-10 parts of a bactericide, 11-25 parts of an energy substrate, and 40-50 parts of a carrier. According to the feed additive, fatty acid is provided through edible oil, the energy substrate provides energy for intestinal epithelial cells, combined with the anti-oxidation effect of the antioxidant and the bactericidal effect of the bactericide, the raw materials are together used for improving the anti-oxidation ability of small intestinal epithelial cells of piglets with intrauterine growth retardation, and then the intestinal immunity and barrier function of piglets with intrauterine growth retardation are further improved.

The invention discloses a method for preventing intestinal radiation damage, wherein the intestinal stem cell genes Dclk1, Msi1, Lgr5, Bmi1 and Notch1 are up-regulated through TPA, the key signal proteins PKC-betaII and Erk1 / 2 as well as the protein p90RSK are further up-regulated, and the intestinal epithelial cell proliferation is promoted to prevent intestinal radiation damage.

The invention provides a preparation method and application of collagen peptide chelated calcium microspheres, belonging to the field of marine biological products, adding CaCl to fish skin collagen solution 2 solution to obtain a collagen peptide chelated calcium solution; the prepared collagen peptide chelated calcium solution and sodium alginate solution were formulated into a mixed solution at a volume ratio of 1:1, and CaCO 3 The suspension was mixed with the mixed solution, and after stirring evenly, it was dropped into the newly prepared D-gluconolactone solution drop by drop with a stainless steel needle, and stirred until a uniform sol was formed; the microspheres were washed and filtered with distilled water, and finally used The filter paper absorbs the moisture on the surface of the microspheres and dries to obtain the microspheres. The microspheres prepared by the present invention hardly swell when passing through the mouth and stomach, but begin to swell when passing through the slightly alkaline distal ileum, and specifically release a collagen peptide to chelate calcium in a short period of time. Chelated calcium is easily absorbed by small intestinal epithelial cells, significantly improving the absorption rate of calcium.

The invention discloses a hesperetin compound with high oral bioavailability and antioxidant activity, and a preparation method and application of the hesperetin compound. In the hesperetin compound, hesperetin is used as a raw material medicine, and TPGS is used as a carrier auxiliary; the mass ratio of hesperetin to TPGS is 1 to (0.1 to 20). TPGS is an effective P-glycoprotein inhibitor which inhibits the drug efflux function of P-glycoprotein; hesperetin is used as a substrate of P-glycoprotein. The hesperetin compound disclosed by the invention can increase the oral bioavailability through the inhibition effect of TPGS on P-glycoprotein of intestinal epithelial cells.

The invention provides a PGC promoter expressed by pig small intestinal epithelial cells and its application. The nucleotide sequence of the PGC promoter is shown in SEQ ID No.1. The present invention predicts the PGC reliable promoter region through an online website; chemical synthesis method Synthesize primers; insert the PGC promoter sequence into the pGL3-Basic vector through the restriction enzyme cutting sites MluⅠ and NheⅠ to obtain the PGC-dual luciferase recombinant plasmid; transfect E. coli for plasmid amplification, extract the plasmid, and verify it by sequencing; Transfected into porcine small intestinal epithelial cells, it was found that the porcine PGC promoter can promote the expression of dual luciferase genes in porcine small intestinal epithelial cells, indicating that the promoter can regulate the expression of downstream genes in porcine small intestinal epithelial cells, which is conducive to the construction of stable The high-efficiency small intestine transcriptional regulatory element has application value in the preparation of transgenic animals and gene therapy research of small intestine diseases.

The invention discloses a preparing method of hydroxyapatite nanoparticles oral system for grafting insulin and gallic acid, comprising the following steps: firstly to synthetize hydroxyapatite (HAP) nanoparticles with microporous morphology, and then tightly wrap HAP surface with PEG, after that, connect gallic acid (GA) to the hydroxyl group of PEG, and finally connect insulin to the PEG surface, to get the nanometer oral transportation system carrying insulin and gallic acid, namely, HAP-PEG- GA-INS nanoparticle. The nano oral transportation system has good stability, natural ingredients, good biocompatibility, high drug loading ratio and release rate, good absorbing effect, the endurance capacity is strong in the gastrointestinal fluid, and absorption is significant in the small intestine epithelial cell line, low cytotoxicity, with significant hypoglycemic effect through oral administration, and has good application potential on diabetes treatment and drug research.