10 results about "Colorectal tissue" patented technology

The present application relates to a kind of anaphalis contorta exosome extraction method and its application.The exosome is obtained by ultra-high speed centrifugation method from fresh anaphalis contorta, particle size is 160-200 nm, Zeta potential is-21.63±1.34 mV, with double membrane structure, mainly containing protein (such as resveratrol O-methyltransferase etc.) and phospholipid, polysaccharide content is only 12.5% of anaphalis contorta fresh medicine, and does not contain traditional active ingredient anaphalis contorta glycoside.Experiments show that anaphalis contorta exosome can be targeted to enrich in colorectal tissue, significantly improve AOM / DSS induced colorectal cancer model mouse tumor microenvironment, reduce tumor number, regulate T cell and macrophage ratio, and relieve pathological damage, better than anaphalis contorta fresh medicine.The mechanism is related to protein component, not polysaccharide or small molecule component.The anaphalis contorta exosome provided in the present application has simple preparation process, high safety, and has significant anti-colorectal cancer application potential.

The invention belongs to the field of biological medicines, and particularly discloses an identification method of honeysuckle flower polysaccharide and application of the honeysuckle flower polysaccharide in preparation of medicines for preventing or treating inflammatory bowel diseases. A DSS solution is used for inducing a mouse inflammatory bowel disease model, honeysuckle polysaccharide FLP is used for gavage treatment, and mouse weight, disease activity indexes, colorectal tissue pathological structures, spleen pathological structures, HE staining, intestinal microbial structures, intestinal metabolite spectrums and the like are detected and analyzed. Results show that the honeysuckle flower polysaccharide can delay inflammatory bowel diseases of model animals in an oral administration manner, and has the characteristics of high targeting property, convenience in administration and the like. Good application prospects are realized in the aspect of treating the inflammatory bowel disease.

This invention discloses a method for culturing organoids from normal colorectal tissue and colorectal cancer, comprising: taking tissue samples from normal colorectal tissue and colorectal cancer, and washing them; cleaning the tissue samples, cutting them into small pieces, and placing them in a buffer solution; removing the supernatant after the tissue samples have settled; adding a dissociation reagent, using a homogenizer to dissociate the tissue, and centrifuging to remove the supernatant; adding a washing buffer, refluxing to release crypts, and obtaining a tissue fragment suspension; filtering to obtain a crypt suspension, centrifuging to remove the supernatant; resuspending the precipitate with a washing buffer to obtain a culture suspension; mixing the matrix gel with the culture suspension to obtain a mixture, and counting crypts; seeding the mixture onto a multi-well plate, incubating, adding organoid culture medium, and culturing to obtain the organoids. This method improves crypt integrity and yield by determining the size of tissue fragments and using a homogenizer in the tissue dissociation process, resulting in a high success rate of organoid culture and good passage stability.

The invention discloses a tissue resident bacterium marker for identifying normal colorectal tissue, polyp and intestinal cancer and application of the tissue resident bacterium marker. According to the present invention, the tissue resident bacterium markers comprise Pseudomonas, Pigmentiphaga, Fusobacteria, Parabacteroides, Bacteroides, Collinsella and Parvimonas, and can be used to reflect the biological specificity of patients, and the biological specificity of the patients can be represented by the tissue resident bacterium markers, such that the biological specificity of the patients can be represented by the tissue resident bacterium markers, and the biological specificity of the patients can be represented by the tissue resident bacterium markers, such that the biological specificity of the patients can be represented by the tissue resident bacterium markers, and the biological specificity of the patients can be represented by the tissue resident bacterium markers; according to the present invention, with the combination model of the six bacteria such as Pseudomonas, Pigmentiphaga, Fusobacteria, Parabacteroides, Bacteroides and Collinsella, the normal tissue and polyp of the colorectum can be accurately identified, and the accuracy is improved by 12.5% compared with the accuracy of the existing non-invasive screening method; the polyp and the intestinal cancer can be more accurately identified by a combined model of four bacteria, namely, Pseudomonas, Pigmentiphaga, Fusobacteria and Parvimonas, and the accuracy of the method is improved by 15.3% compared with that of an existing non-invasive screening method. Moreover, the invention solves the problem of sample heterogeneity in biopsy-based diagnosis, can be expected to be used for guiding clinical diagnosis and treatment, and provides a new way for early discovery and early treatment of polyp or intestinal cancer.

The invention discloses a nano-drug system for targeted therapy of colorectal cancer with fusobacterium nucleatum infiltration and a preparation method of the nano-drug system. The invention belongs to the technical field of biology, and particularly relates to a nano-drug system for targeted therapy of colorectal cancer with fusobacterium nucleatum infiltration and a preparation method of the nano-drug system. An active ingredient of the medicine with the tumor cell activity inhibiting effect or the anti-tumor activity contains NM-coated PLGA-MTI-OXA, the NM-coated PLGA-MTI-OXA is composed of neutrophile granulocyte membrane vesicles and PLAG-MTI-OXA, the PLAG-MTI-OXA is nanoparticles obtained by loading chemotherapeutic drugs MTI and OXA on polylactic acid-glycolic acid copolymer, and the neutrophile granulocyte membrane vesicles are derived from bone marrow neutrophile granulocytes. The NM-coated PLGA-MTI-OXA developed by the invention can effectively target colorectal tissues infiltrated by fusobacterium nucleatum, successfully realizes antibacterial and anti-tumor effects, and has application value.

The present invention discloses a dissociation kit for mouse colorectal single cell suspension, which is composed of the following components: a first enzymatic digestion solution, a second enzymatic digestion solution, a third enzymatic digestion solution, a PBS buffer solution, an RPMI-1640 culture medium, and an erythrocyte lysate; the first enzymatic digestion solution is a 20 mM EDTA solution; the second enzymatic digestion solution is a 0.25% trypsin-EDTA solution; the concentration of neutral protease in the third enzymatic digestion solution is 100 mg / mL, the concentration of collagenase I is 100 mg / mL, and the concentration of collagenase II is 100 mg / mL; a method for extracting mouse colorectal single cell suspension using the dissociation kit is also disclosed. The reagent materials used in the dissociation solution in the kit are easy to obtain, contain no harmful components, are safe and environmentally friendly, the method is simple to operate, and the final result after the cooperation of each step has high repeatability and success rate, and can stably obtain a mouse colorectal single cell suspension with high viability and low impurities.

The invention relates to an extraction method and application of an anoectochilus formosanus exosome. The exosome is separated from fresh anoectochilus formosanus through an ultra-high-speed centrifugation method, the particle size is 160-200 nm, the Zeta potential is-21.63 + / -1.34 mV, the exosome is of a double-layer membrane structure and mainly contains protein (such as resveratrol O-methyltransferase) and phospholipid, the polysaccharide content is only 12.5% of that of fresh anoectochilus formosanus medicine, and the exosome does not contain traditional active ingredient kinsenoside. Experiments show that the anoectochilus formosanus exosome can be enriched in colorectal tissues in a targeted manner, the tumor microenvironment of AOM / DSS induced colorectal cancer model mice is remarkably improved, the number of tumors is reduced, the ratio of T cells to macrophages is adjusted, pathological injuries are relieved, and the effect is better than that of an anoectochilus formosanus fresh medicine. The mechanism is related to protein components instead of polysaccharide or small molecule components. The anoectochilus formosanus exosome provided by the invention is simple in preparation process and high in safety, and has remarkable anti-colorectal cancer application potential.

The invention discloses a colon targeted delivery system as well as a preparation method and application thereof. The colon targeted delivery system comprises amifostine, pectin and chitosan, the preparation method comprises the following steps: carrying out chemical amidation reaction on pectin and amifostine, connecting carboxyl in the pectin with amino in the amifostine through an amido bond to form a nanoparticle core PEC-AMF, and carrying out electrostatic interaction on chitosan and the pectin to form a nanoparticle wrapping layer to synthesize CS / PEC-AMF nanoparticles; according to the colon targeted delivery system, after oral administration, accurate and controlled drug release is realized by virtue of a pH value which changes step by step in a gastrointestinal tract environment and a specific pectinase catalysis effect in colon; experiments prove that the delivery system has excellent stability in the stomach and the small intestine, and can quickly release amifostine at the colon part, so that the intestinal epithelium and immune cells are effectively protected from damage caused by radiotherapy, and meanwhile, pectin regulates the intestinal microecology and protects normal tissues of colorectum without influencing the effect of radiotherapy on tumors.

The invention relates to the technical field of biomedical engineering, and discloses EO-CRC-sourced NCIO and a culture method and application thereof.The method comprises the steps that normal colorectal tissue of a premature colorectal cancer patient with the onset age smaller than 50 years old is obtained; digesting the tissue blocks for 20-60 minutes at 2-6 DEG C by using EDTA (Ethylene Diamine Tetraacetic Acid) digestive juice; stopping the digestion reaction, filtering, and centrifugally collecting a digestion product; mixing the collected digestion product with a basic culture medium and matrigel in proportion, and performing three-dimensional embedding; and after gelling, adding a normal intestinal organ-like culture medium for culturing. According to the method, culture conditions are optimized according to biological characteristics of normal tissues of a patient suffering from premature colorectal cancer, and cell damage is avoided by adopting EDTA low-temperature digestion. The cultured organoid can be stably passed for more than 15 generations, and has important application value in the fields of premature colorectal cancer disease model construction, pathogenesis research, drug screening and the like.

The utility model discloses a colorectum fixing and pulling-out auxiliary device which comprises two sets of limiting and fixing plates, the limiting and fixing plates are in an arc shape, a semicircular notch is formed in one side of each limiting and fixing plate, the two sets of limiting and fixing plates are spliced to limit the colorectum through the notches, containing grooves are formed in the two ends of each limiting and fixing plate, and the limiting and fixing plates are arranged in the containing grooves. Dirty liquid generated by an operation can slide into the containing grooves in the two ends through the surface of the arc-shaped limiting fixing plate to be collected, the dirty liquid is discharged through the drain outlets in the bottoms of the containing grooves, and limiting mechanisms are arranged at the two ends of the limiting fixing plate. According to the colorectum fixing and pulling-out auxiliary device, the two sets of limiting and fixing plates are matched with the notches to limit and fix the colorectum, medical staff can carry out resection operation on the colorectum of the limiting part, dirty liquid generated in the resection process can slide towards the two ends along the surfaces of the limiting and fixing plates and is collected in the containing grooves, and therefore the medical staff can carry out resection operation on the colorectum. And infection caused by backflow of dirty liquid into the abdominal cavity can be effectively avoided.