12 results about "Renal epithelial cell" patented technology

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The application discloses immortalized chicken embryo epithelioid cells and a preparation method and application thereof, and the preparation method of the immortalized chicken embryo epithelioid cells comprises the following steps: extracting primary chicken embryo epithelial cells from a chicken embryo; and culturing the primary chicken embryo epithelioid cells in a conditional medium containing chicken Noggin protein for subculture, so as to obtain the immortalized chicken embryo epithelioid cells. The immortalized chicken embryo epithelioid cells prepared by the preparation method can be stably subcultured for a long time, can maintain stable proliferation ability after multiple subcultures, reduces individual differences, and improves the reliability and repeatability of experimental results. In addition, the preparation method does not introduce exogenous genes or change the genotype of the cells, ensures the stability at the gene level, and avoids the interference of the exogenous genes on the stability of the cell phenotype.

The invention relates to the technical field of medicine synthesis, in particular to ergosterol amino alcohol derivatives with antibacterial activity and a preparation method and application thereof. The structure is as shown in general formula (I). The ergosterol amino alcohol derivative disclosed by the invention is high in gram-positive bacterium and gram-negative bacterium resisting activity, and the antibacterial activity of the derivative with the optimal activity on staphylococcus aureus and escherichia coli is improved by 64 times and 32 times respectively, so that the ergosterol amino alcohol derivative is obviously superior to that of lead compound ergosterol and compounds reported in literatures. The IC50 values of the human body normal cells HK-2 (near-end renal tubular epithelial cells) and HEK-293 (human embryo renal epithelial cells) are obviously higher, and when MIC < lt > is less than MIC < lt >, MIC < lt > is less than MIC < lt >; the ergosterol amine alcohol derivative has the selectivity index SI of more than or equal to 6.0 when the molecular weight of the ergosterol amine alcohol derivative is 32 [mu] g mL <-1 >, has the characteristics of high efficiency and low toxicity, and has potential application value in the aspect of anti-infection drug development.

The invention discloses a stably transfected cell strain for detecting activity of Wnt and / or R-spondin family cytokines as well as a construction method and application of the stably transfected cell strain, and belongs to the technical field of biological medicines. Comprising the following steps: preparing target plasmids: respectively connecting a SuperTOP sequence and a SuperFOP sequence to two different expression plasmids, and respectively connecting luciferase reporter genes to obtain a target plasmid A and a target plasmid B; respectively transfecting the target plasmid A and the target plasmid B into human kidney epithelial cells, culturing in a puromycin-containing culture medium, and sorting positive cells according to labeled fluorescence GFP (Green Fluorescent Protein) to obtain stably transfected cells A and stably transfected cells B. The method has the advantages that activity detection of Wnt family and R-spondin family cell factors is carried out on stably transfected cells, operation is simple, the detection period is shortened by 25%, sensitivity is improved by at least 50%, and the method has high application prospects and economic value.

The present application relates to the technical field of genetic engineering, and specifically discloses a 293T cell line stably expressing ruminant SLAM protein, a construction method and application thereof, which comprises introducing an HA tag at the 3' end of a ruminant SLAM gene to obtain a SLAM-HA target gene, connecting the SLAM-HA target gene to a lentivirus vector CD513B-Cherry, transforming DH5a competent cells, extracting a positive plasmid to obtain a lentivirus vector plasmid; mixing the lentivirus vector plasmid with an auxiliary plasmid, and respectively transfecting 293T cells to obtain packaged SLAM lentivirus; and transducing the SLAM lentivirus into human renal epithelial cells to obtain a human renal epithelial cell line stably expressing ruminant SLAM protein. The present application successfully constructs a cell line stably expressing ruminant SLAM protein, which can obviously mediate the infection and proliferation of PPRV, and has potential for preparing a vaccine.

The application discloses application of seleniumized Chinese herbal medicine polysaccharide in preparation of preparations for preventing and treating calculus. The seleniumized corn silk polysaccharide and the seleniumized desmodium polysaccharide are obtained by extracting corn silk polysaccharide and desmodium polysaccharide and seleniumizing the same. It is found through detection that the seleniumization of the corn silk polysaccharide and the desmodium polysaccharide improves the protection of the corn silk polysaccharide and the desmodium polysaccharide on renal epithelial cells, so that the corn silk polysaccharide and the desmodium polysaccharide can not only effectively inhibit the oxidative damage of renal epithelial cells induced by oxalic acid, but also effectively inhibit the adhesion of calcium oxalate crystals on the renal epithelial cells, and the generation of calculus can be reduced from two aspects of protecting cells and inhibiting the adhesion of calcium oxalate crystals. In addition, it is also found that the combination of the seleniumized corn silk polysaccharide and the seleniumized desmodium polysaccharide can further improve the effect of preventing and treating calculus, has obvious synergistic effect, and is helpful to the prevention and treatment of calculus.

According to the laminaria proteoglycan purified extract and the preparation method and application thereof, in the preparation process of the purified extract, an ultrasonic-assisted water extraction method and anion exchange chromatography are adopted for purification, the extraction efficiency is high, the purification effect is good, laminaria proteoglycan effective ingredients in laminaria can be effectively enriched, and the content of laminaria proteoglycan in laminaria is increased. The obtained extract is high in purity and high in activity, high-value utilization of kelp resources is achieved, kelp proteoglycan is used as a whole for extraction, purification and activity research, and the synergistic effect of kelp proteoglycan is fully exerted. In the invention, a caenorhabditis elegans model, a zebra fish model and a human renal epithelial cell (HK-2) model verify that the laminarin purified extract has a remarkable protective effect on the body injury caused by the microplastics, and a new candidate medicine and a scientific basis are provided for developing the microplastic injury protection medicine.

This invention provides a high-throughput detection method for membrane protein-binding protein affinity, comprising: S1. Designing and constructing a binding protein expression vector; S2. Binding protein preparation and pretreatment: transfecting the binding protein expression vector with a human embryonic kidney epithelial cell line, and collecting the cell supernatant containing the binding protein after transfection; S3. Establishing a co-incubation system between the binding protein and target cells: co-incubating the cell supernatant containing the binding protein with test target cells to allow the membrane protein on the target cells to bind to the binding protein, followed by washing to obtain target cells bound with the binding protein; S4. Luciferase activity detection: adding the luciferase nano-substrate Furimazine to the target cells bound with the binding protein for quantitative detection of luciferase activity; S5. Evaluating the affinity between the membrane protein on the target cells and its binding protein. This invention uses the natural membrane environment of living cells as the reaction matrix, enabling rapid and high-throughput evaluation of the strength of cell membrane protein-binding protein affinity, with a simplified process and lower cost.

The invention belongs to the technical field of biological medicine, and discloses a carbazole compound embedded with a boron-nitrogen unit, and a synthesis method and application thereof, the carbazole compound has a structure of a general formula (I): (A)-(L)-(B), including isomers or pharmaceutically acceptable salts thereof. The synthesis method of the compound is simple and convenient to operate, mild in condition and wide in substrate applicability. The PARP14 inhibitor GeA-69 is subjected to BN modification through the method, and the obtained compounds G-1 and G-2 have low toxicity to normal cells HEK-293 (human kidney epithelial cell line) while keeping the inhibitory activity to tumor cells such as HeLa cells (cervical cancer cell line) and the like. The invention provides a new strategy for development of new drugs based on a carbazole structure.

This invention provides the application of an S100A6 inhibitor in the preparation of a drug for treating calcium oxalate stones. This invention is the first to discover that S100A6 plays a crucial role in the formation of calcium oxalate stones. It promotes STAT3 phosphorylation, upregulates the expression of NADPH oxidase subunits p67phox and p40phox, promotes reactive oxygen species (ROS) generation, and induces autophagy and apoptosis in renal epithelial cells, thereby promoting calcium oxalate stone formation. Furthermore, the S100A6 small molecule inhibitor AP-048 provided by this invention specifically binds to S100A6. In vitro and in vivo experiments have confirmed that AP-048 can significantly inhibit calcium oxalate crystal cell adhesion, reduce ROS levels, decrease renal crystal deposition, alleviate renal tubular damage and inflammatory factor expression, and has no significant hepatotoxicity or nephrotoxicity. This invention provides a new target and therapeutic strategy for calcium oxalate stones, with promising clinical application prospects.

The invention discloses an immortalized chick embryo kidney epithelioid cell as well as a preparation method and application thereof. The preparation method of the immortalized chick embryo kidney epithelioid cell comprises the following steps: extracting a primary generation chick embryo kidney epithelioid cell from a chick embryo; and placing the primary chicken embryo kidney epithelioid cells in a conditioned medium containing chicken Noggin protein for subculture to obtain the immortalized chicken embryo kidney epithelioid cells, the immortalized chicken embryo kidney epithelioid cell prepared by the preparation method can be stably passaged for a long time and can still keep stable multiplication capacity after multiple passations, individual differences are reduced, the reliability and repeatability of experimental results are improved, in addition, the preparation method does not introduce exogenous genes or change genotypes of the cells, and the preparation method is simple and convenient to operate. The stability on the gene level is ensured, and the interference of exogenous genes on the cell phenotype stability is avoided.

The application provides a laminarin proteoglycan purified extract and a preparation method and application thereof. In the preparation process of the purified extract, ultrasonic-assisted water extraction and anion exchange chromatography are used for purification, the extraction efficiency is high, the purification effect is good, the effective components of laminarin proteoglycan in laminaria can be effectively enriched, the obtained extract has high purity and strong activity, the high-value utilization of laminaria resources is realized, the laminarin proteoglycan is extracted and purified as a whole and activity research is conducted, and the synergistic effect is fully exerted. In the application, through the Caenorhabditis elegans model, the zebra fish model and the human kidney epithelial cell (HK-2) model, it is verified that the laminarin proteoglycan purified extract has a significant protection effect on the damage of the organism caused by microplastics, and a new candidate drug and scientific basis are provided for the development of a microplastic damage protection drug.