37 results about "Merokeratin" patented technology

The invention provides a reagent for diagnosing cervical cancer, which can not only detect the presence or absence of cervical cancer but can also distinguish squamous cell carcinoma and adenocarcinoma from each other, a method for screening cervical cancer by using the reagent, particularly a screening method capable high speed processing by utilizing flow cytometry. The reagent comprises a first labeled antibody reacting with gland cells, a second labeled antibody reacting with adenocarcinoma cells and a third labeled antibody reacting with atypical cervical squamous cells, the antibodies being labeled with mutually distinguishable labels respectively. Preferably, at least one selected from the group consisting of MUC1 antibody, cytokeratin 7 antibody, and cytokeratin 18 antibody is used as the first labeled antibody, at least one selected from the group consisting of cytokeratin 8 antibody and HIK1083 antibody is used as the second labeled antibody, and at least one member selected from the group consisting of NMP179 antibody, p16[INK4A] antibody, Ki-67 antibody, p53 antibody, p21 antibody, EMA antibody, CEA antibody and MIB-1 antibody is used as the third labeled antibody.

The invention relates to a quantum dot fluorescent probe and application thereof. The invention is characterized in that the quantum dot fluorescent probe utilizes a difunctional cross-linking agent long-chain succinimidyl-4-[N-maieimidomethyl]cyclohexane-1-carboxy-[6-amidocaproate] (SMCC) to undergo a cross-linking reaction with an amino group on the surface of an amino quantum dot; a maleimide group is produced on the surface of the quantum dot; a to-be-labeled antibody is reduced by a reducing agent dithiothreitol (DTT) so as to reduce a disulfide bond into a mercapto group; and the mercapto group and the maleimide group form a covalent bond so as to realize labeling of the quantum dot with the antibody. The quantum dot fluorescent probe is applied to (1) detection of one protein oncofetal antigen or (2) detection of a plurality of protein CEA, euron-specific enolase (NSA) and a cytokerain fragment 19CYFRA21-1, wherein the detection limit of detection (1) is 38 pg / ml, and the detection limit of detection (2) is 0.9 ng / ml.

The present invention relates to enrichment and / or identification of fetal cells of a maternal blood sample using fetal cell specific ligands and / or fetal cell specific hybridization probes wherein the ligand or probes are directed to an endothelial / mesenchymal marker, e.g. CD105, CD146 or CD141, in a first round of enrichment and the ligand or probes, in a second round of enrichment, are directed to an epithelial marker, e.g. a cytokeratin, such as CK7, CK8, CK18 or CK19. Enriched or identified fetal cells may be subjected to steps of detection or diagnosis, wherefore the present invention enables non- invasive 5 prenatal diagnostics.

The invention relates to a plate-type chemiluminescence detection kit of a cytokeratin 19-fragment. The kit comprises the following reagents: biotin-marked CYFRA21-1 antibody solution, an elisa plate coated by anti-biotin, horse-radish-peroxidase-marked CYFRA21-1 antibody solution, a CYFRA21-1 calibration product A-F, concentrated washing liquor, substrate solution A, and substrate solution B. The main design idea of the detection kit is that the detection specificity of the kit is stabilized by introducing a biotin system with the anti-biotin on the basis of chemiluminescence immune assay, and the detection sensitivity and detection time are greatly increased. A high-temperature oscillatory-type anti-biotin coating technology is improved, so the usage amount of antibodies / antigens is reduced, the production flow of the reagents is simplified, and the production cycle is shortened.

Carcinomas typically invade as a cohesive multicellular unit, a process termed collective invasion. It remains unclear how different subpopulations of cancer cells contribute to this process. We developed three-dimensional (3D) organoid assays to identify the most invasive cancer cells in primary breast tumors. Collective invasion was led by specialized cancer cells that were defined by their expression of basal epithelial genes, such as cytokeratin-14 (K14) and p63. Furthermore, K14+ cells led collective invasion in the major human breast cancer subtypes. Importantly, lumenal cancer cells were observed to convert phenotypically to invasive leaders following induction of basal epithelial genes. Although only a minority of cells within lumenal tumors expressed basal epithelial genes, knockdown of either K14 or p63 was sufficient to block collective invasion. Our data reveal that heterotypic interactions between epithelial subpopulations are critical to collective invasion. We suggest that targeting the basal invasive program could limit metastatic progression.

The invention relates to a detection kit for peripheral blood circulating tumor cells of small cell lung cancer patient and PD-L1, and belongs to the technical field of peripheral blood circulating tumor cell detection reagents. A reagent for circulating tumor cell detection comprises a CTC detection primary antibody mixture and a CTC detection secondary antibody mixture, wherein the CTC detectionprimary antibody mixture comprises Anti-Cytokinase, anti-CD45, anti-pan keratin, anti-EpCAM and a Pears immunopotentiator, and the CTC detection secondary antibody mixture comprises goat anti-mouse AlexaFluor 488, goat anti-rabbit AlexaFluor 647, a DAPI working solution and a Pears immunopotentiator. The reagent provided by the invention can greatly improve the detection efficiency, reduce the detection cost and improve the detection specificity and accuracy.

Methods and compositions for detecting tumor origin are disclosed. In certain embodiments, the origin is determined by detecting expression levels of pan-epithelial membrane mucin (MUC1), intestinal-type secretory mucin (MUC2), cytokeratin 17 (CK17), or a combination thereof. Also disclosed are methods for aiding in the determination of an appropriate course of treatment for a subject with a tumor, or for predicting a therapeutic outcome of a subject, based on the expression levels of MUC1, MUC2, CK17, or a combination thereof. Kits for use in each of these methods are also provided.

The invention provides a breast cancer cell dyeing method. The method takes anti-pan cytokeratin antibody (AE1 / AE3) as the primary antibody, and adopts a horseradish peroxidase labeled rabbit antimouse antibody as the second antibody to label breast cancer cells, by means of diaminobenzidine (DAB) and hematoxylin dyeing, breast cancer cells can be dyed specifically. The invention also puts forward application of the dyeing method and a kit for carrying out the method. The method provided by the invention has the advantages of simple operation and good accuracy, can be used for detection of breast cancer micrometastatic cells in bone marrow blood and peripheral blood samples of breast cancer patients, and has important reference significance to prognosis of breast cancer.

The invention discloses a feed additive for improving the yield and quality of sheep wool, feed and a preparation method. The additive comprises processed radix aconiti lateralis, Chinese dates, loofah sponge, kudzu vine root, common bombax flowers, asarum sieboldii, gynostemma pentaphylla, holy basil, fructus aurantii and dried ginger. The feed additive takes the radix aconiti lateralis, the Chinese dates, the loofah sponge, the kudzu vine root, the common bombax flowers, the asarum sieboldii, the gynostemma pentaphylla, the holy basil, the fructus aurantii and the dried ginger as raw materials, by using the synthetic effects of the materials, rich nutrients are provided for sheep, blood circulation of the sheep and metabolism of epithelial cells are promoted, synthesis of wool keratin ispromoted, the growth speed and yield of the wool are improved, and the wool is slender and white. Meanwhile, the traditional Chinese medicine components have the effects of clearing away heat and toxic materials, tonifying middle-jiao and qi, nourishing blood and soothing the nerves and the like, can improve the immunity of sheep bodies, absorption and utilization of the effective components suchas nutritional ingredients and microelements in the feed for the sheep are improved, the growth speed and yield of the wool are further increased, and the economic benefit of sheep breeding is greatly improved.

The invention discloses a method for preparing double-resistant sandwich method test paper for ancient wool fabric. The method comprises the following steps that gold nano particles are prepared through a method for reducing chloroauric acid through sodium citrate; a Jinbiao rabbit anti-keratin antibody is prepared; test strips are assembled, a sample pad, a colloidal gold alloy pad, a nitrocellulose membrane and a water absorbing pad are sequentially assembled on a PVC bottom plate and cut into the test paper strips through a cutter, and the test strips are placed in an aluminum foil bag to be sealed and stored; 1 g of fabric trace sample is taken to be dissolved in 50-100 ml of Tris-HCl buffer liquid with the PH of 8.2, even stirring is carried out, a drop of supernate is taken to drop onto the sample pad of the assembled test strips after 2 h; after 5-10 min, a detecting line and a quality control line both display red, and it is indicated that keratin is included in the sample, and the fabric trace is the trace of the wool fabric. Compared with the prior art, the method is simple and fast, high in sensitivity, high in specificity, visual in result and more suitable for archaeology field analysis.

Disclosed is a method of amplifying and detecting cytokeratin 19 mRNA in RNA amplification process, comprising: a step for forming a double-stranded DNA containing a promoter sequence with a reverse transcriptase by use of a combination of oligonucleotides consisting of a first primer having a sequence homologous to a portion of cytokeratin 19 mRNA and a second primer having a complementary sequence, wherein the promoter sequence is added to the 5′-end of either the first primer or the second primer, forming an RNA transcription product by use of an RNA polymerase with using the double-stranded DNA as template, and forming the double-stranded DNA by use of a reverse transcriptase by continuing to use the RNA transcription product as a template of DNA synthesis, by measuring an amount of amplified RNA produced over time with an oligonucleotide probe designed so that signal properties change with the formation of a complementary double strand with the amplified RNA.

The invention relates to a CK20 antigen, a hybridoma cell strain, a monoclonal antibody, and application thereof. High-variation head and tail sequences of CK20 are selected according to the characteristics of a cytokeratin sequence and connected by adopting connecting components, gene synthesis is carried out according to the sequence of head-connecting-components-tail, and a prokaryotic expression vector is inserted, expressed and purified to obtain the CK20 antigen; and the hybridoma cell strain C7C11 capable of stably secreting the mouse anti-human CK20 monoclonal antibody is finally obtained by using the CK20 antigen as an immunogen to immunize a BALB / c mouse, fusing spleen cells of the mouse with SP2 / 0 cells, obtaining monoclonal cells by using a limited dilution method and screeningpositive hybridoma cells by using an indirect ELISA method, and the preservation number is CCTCC NO: C2020201. The monoclonal antibody is prepared through induction in an animal body, and the specificity and the sensitivity of the monoclonal antibody are verified through Western Blot and immunohistochemistry (IHC). The CK20 monoclonal antibody provided has better specificity and sensitivity, andcan be used for preparing a CK20 in-vitro immunoassay reagent or a kit.

The invention provides a breast cancer sentinel lymph node metastasis in-vivo fluorescence targeting tracer agent and a preparation method thereof. The targeting tracer agent comprises a cytokeratin 19 monoclonal antibody and Cy754, epsilon-amino of lysine in the cytokeratin 19 monoclonal antibody and carboxyl of the Cy754 form amido bonds to be combined, and one CK19 monoclonal antibody moleculeis connected to 8-12 Cy754 molecules. The preparation method comprises the following steps: adding 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride into a mixed solution of the CK19 monoclonal antibody and Cy754, carrying out a reaction, and carrying out dialysis purification on the material obtained after the reaction to obtain the tracer agent. According to the tracer agent providedby the invention, when the region is detected through the photoacoustic conversion detector, the lymph node subjected to fluorescence targeting development can be found, so that the purpose of determining sentinel lymph node metastasis in vivo is achieved.

The invention discloses a method for improving the enzymatic modification efficiency of wool keratin, which belongs to the technical field of biochemical modification of textile materials. By utilizing the characteristics of disulfide bonds rich in a structure of the wool keratin, an amino group is introduced into the structure of the keratin by performing a click chemical reaction on a sulfhydrylgroup formed by reduction and propargylamine, so that the number of acting sites of a TGase enzymatic reaction is increased, and the enzymatic modification efficiency of the wool keratin is improved.The method is high in operating reaction efficiency, has no toxicity and is easily controlled, and the problems of residual of by-products, environmental pollution, fabric strength damage and the like caused by a side reaction of the chemical reaction, are avoided, therefore, the method has strong practical value.

The invention discloses a cervical cancer screening method, which comprises the following steps of: taking menstrual blood, and detecting CA15-3(Carbohydrate antigen 15-3, carbohydrate antigen 15-3), CEA (Carcino-embryonic antigen), CA125 (Carbohydrate antigen 125), CYFRA21-1 (cytokeratin 19 fragment, cytokeratin fragment), squamous cell carcinoma associated antigen (SCCA), Her-2 (human epidermal growth factor receptor-2 of protooncogene), TP53 (Tumor protein 53) gene, APC (Adenomatous polyposis coli) gene, HPV DNA gene, and TCT(liquid based cytology) detection in the menstrual blood, and the illness risk of cervical cancer can be prompted. The method has no radiation damage to human bodies, and is short in detection time and low in cost. As the menstrual blood is very convenient and noninvasive to obtain, women can detect once a month, so that the women can know the risk of cervical cancer as soon as possible and take prevention and treatment measures.

The invention belongs to the technical field of biomedical detection, and particularly relates to a marker and a detection kit for detecting bladder cancer. The marker provided by the invention comprises a nuclear matrix protein 22 (NMP22), a carcino-embryonic antigen (CEA), a bladder tumor antigen (BTA) and a cytokeratin 19 fragment (CYFRA21-1). According to the detection kit, acridinium ester is adopted for direct chemiluminescence, an antibody (antigen) is directly labeled by the acridinium ester, after immune reaction with a corresponding antigen (antibody) of a sample to be detected, a coated antibody-antigen to be detected-acridinium ester labeled antibody compound is formed, excitation liquid is added, and decomposition and luminescence can be realized without a catalyst. A combination of multiple tumor markers is established, the detection specificity and sensitivity of bladder cancer are improved through combined detection of multiple indexes, when urine of a bladder cancer patient is detected, the sensitivity and specificity are high, and early screening, treatment effect and postoperative recurrence of the bladder cancer patient can be dynamically monitored.

Provided are a pretreatment method for detecting the number of cells containing Ki-67 protein-positive nuclei using Ki-67 antibody; a method for the detection; a kit to be used in the detection method; and determination of a therapy regimen using the aforesaid method. Attempts were made to activate Ki-67 antigen with the use of an enzyme having been considered as inappropriate for the activation thereof. By pretreating a sample with an enzyme not recognizing the epitope of MIB-1 (a rare cutter enzyme), the antigen activation of Ki-67 antigen was enhanced, while enhancing the antigenicity of other antigens including a cytokeratin too. As a result, a method for more objectively and more universally quantifying Ki-67-positive cells at higher reproducibility, said method comprising isolating cell nuclei from an FFPE section while enhancing the antigenicity and performing a reaction between Ki-67 protein, i.e., the target, existing in the cell nuclei with a fluorescently labeled antibody, has been completed.

The invention relates to a cytokeratin 18 gene expression detection kit and a detection method thereof. The cytokeratin 18 gene expression detection kit comprises a capture probe aiming at CK18 gene mRNA and a signal amplification system, the capture probe is a stem-ring-shaped gamma PNA-PNA probe, which sequentially comprises a stem structure sequence, a P1 sequence, a Linker sequence and a P2 sequence from the 5' end to the 3' end; and the signal amplification system comprises a marking probe SP1 and a marking probe SP2, wherein each labeled probe SP1 sequentially comprises a P3 sequence, a spacer sequence and a P4 sequence from the 5' end to the 3' end; the 5' end of the P3 sequence is modified with a fluorophore, each labeled probe SP2 sequentially comprises a P5 sequence, a spacer sequence and a P6 sequence from the 5' end to the 3' end, and the 3' end of the P6 sequence is modified with a fluorophore. The kit has the advantages of high sensitivity, strong specificity, high accuracy, short detection time and the like.

The invention discloses a lung cancer risk prediction model based on CT images and biomarker spectrum for Chinese urban population with pulmonary nodules. Phosphate buffer solution with a value of 7.4, serum protein dilution, stop solution, tetramethylbenzidine substrate solution, normal human serum and positive control serum; the biomarkers include gastrin-releasing propeptide, Four biomarkers including carcinoembryonic antigen, fragment of cytokeratin 19 and squamous cell carcinoma antigen were combined with CT imaging parameters and other clinical indicators. The present invention can assist in screening for lung cancer.

The invention belongs to the technical field of biological detection, and discloses a fluorescent quantum dot nanoprobe, a preparation method thereof and application of the fluorescent quantum dot nanoprobe in detection of circulating tumor cells. The detection probe comprises a water-soluble quantum dot modified by an antibody; the antibody can specifically bind to a circulating tumor cell surface marker, such as an anti-EpCAM antibody, an anti-EGFR antibody, an anti-cytokeratin CK-8, CK-18 and CK-19 antibody, an anti-tumor cell glycoprotein (TAG) antibody or an anti-lactoglobulin antibody; more preferably, an anti-EpCAM antibody; the water-soluble quantum dots are selected from near-infrared two-region fluorescent water-soluble non-toxic quantum dots. An antibody-modified water-soluble near-infrared two-region fluorescent non-toxic quantum dot material is used as a fluorescent probe, and the number of circulating tumor cells is detected by virtue of the change of the fluorescence intensity of the probe through an antigen-antibody specific recognition effect.

The invention relates to a preparation method of an electrochemical immunosensor based on a polypyrrole modified silver phosphate loaded silver nanosphere composite material. According to the invention, a primary antibody marker is formed by internal packaging of a Prussian blue analogue (NiFe@FeFe PBA) with a core-shell heterostructure and combination of a primary antibody externally loaded with luminol and a soluble fragment of cytokeratin 19; and a polypyrrole modified silver phosphate loaded silver nanosphere composite material (Ag3PO4@PPy-Ag) is combined with a second antibody of a soluble fragment of cytokeratin 19 to form a second antibody marker, so that the construction process of the sensor is simplified. Luminol is packaged into a porous core-shell heterostructure nanocube NiFe@FeFe PBA, so that the nano composite material with an excellent electrochemical luminescence behavior is obtained. Ag3PO4@PPy-Ag can effectively quench electrochemical luminescence of luminol, a sandwich quenching type electrochemical immunosensor is constructed based on the resonance energy transfer principle, ultra-sensitive detection of soluble fragments of cytokeratin 19 is achieved, and the detection limit is 28.43 fg mL <-1>.

The invention provides a radionuclide targeted breast cancer sentinel lymph node metastasis tracer and a preparation method thereof. The tracer comprises a cytokeratin 19 (CK19) monoclonal antibody and pertechnetate, and the pertechnetate is subjected to a reaction through 2-mercaptoethanol and is coupled to the CK19 monoclonal antibody. The preparation method comprises the following steps of performing a mixed reaction on the CK19 monoclonal antibody with the 2-mercaptoethanol, adding micromolecular dextran to be uniformly mixed, adding sodium glucoheptonate, stannous chloride and sodium pertechnetate for a reaction, and performing separation by adopting a PD-10 column. The tracer prepared by the method can effectively reduce local trauma of a patient by detecting the metastasis conditionof sentinel lymph nodes in vivo, provides a favorable decision basis for surgeons, and shortens the operation time of the patient.

An immortalized sweat gland myoepithelial cell which expresses α-SMA and pan-cytokeratin and has a sphere forming ability after subculture at least 5 times. A method for producing immortalized sweat gland myoepithelial said method comprising: while culturing a cell structure, wherein sweat gland myoepithelial cells are exposed on a surface, in a state of being suspended in a medium, transferring an immortalizing gene into the cells; and then culturing the transgenic structure thus obtained in a state of being suspended in the medium to thereby obtain immortalized sweat gland myoepithelial cells.

The present disclosure relates to a method of identifying an individual having non-small cell lung carcinoma as to be treated by chemotherapy based on marker molecules cytokeratin-19 fragments (CYFRA 21-1) and carcinoembryonic antigen (CEA) as well as the use of the marker molecules for the identification of an individual to be treated by chemotherapy.

The invention provides a method for preparing a porous material on the basis of wool keratin cortex cells, and belongs to the field of preparation of a high molecular material. The method comprises the steps of: firstly, preparing cortex cell aqueous suspension by taking the wool keratin cortex cells as a raw material; then adding glutaraldehyde into the cortex cell aqueous suspension, and carrying out magnetic stirring to obtain uniform milk white suspension; then rapidly placing the milk white suspension at a temperature of minus 40 DEG C to freeze, and then carrying out vacuum freezing drying; and finally, placing a sample after vacuum freezing drying into a baking oven to bake, and enabling the glutaraldehyde and keratin to be cross-linked so as to obtain the wool cortex cell porous material. According to the invention, the porous material with a large gap is prepared with the simple method, can be used in the fields of heat preservation, heat insulation, damping and the like, andbelongs to the technical field of the high molecular material.

The invention relates to a method for obtaining artificial skin through stem cell in-vitro differentiation and application of the method, and provides a method for differentiating stem cells to skin cells in vitro to obtain artificial skin and application. The culture method comprises the following steps: S1, subculturing mesenchymal stem cells until 60-90% of cells are merged, and collecting P3 generation cells to prepare a mesenchymal stem cell suspension; S2, attaching, specifically, placing acellular dermis and the mesenchymal stem cell suspension in an ultralow adsorption reactor, and carrying out culturing by using a complete culture medium; and S3, inducing the mesenchymal stem cells loaded by the acellular dermis to differentiate to skin cells in vitro, specifically, changing liquid by using an induction culture solution, wherein the induction culture solution takes a KFSM culture medium as a basic culture medium, astragalus injection with the concentration of 30-50 g / L and fetal calf serum with the volume fraction of 1-3% are added, and the induction culture time is 14 days. TM4SF1 and broad-spectrum cytokeratin expression are positive, the obtained artificial skin is moresimilar to cells forming skin tissues, and a better treatment effect can be achieved.

The invention provides a marker and kit for early hepatic fibrosis diagnosis and application.The marker comprises at least one of cytokeratin 19 and a fragment antigen CYFRA21-1 of the cytokeratin 19,and at least one of the liver fibrosis early diagnosis markers is differentially expressed in a test sample and a control sample. The marker for early diagnosis of hepatic fibrosis can realize diagnosis of early hepatic fibrosis.

The invention provides a wool keratin cortex cell preparation method, and belongs to the field of wool keratin cortex cell extraction. The method includes the steps: firstly, pretreating wool by calcium hypochlorite / hydrogen peroxide and stripping wool surface scale layers; secondly, taking L-cysteine as a reducing agent, performing trypsin catalysis and degrading wool fibers by CMC (carboxymethylcellulose) to separate wool cortex cells; thirdly, further separating the cortex cells by ultrasonic treatment; finally, filtering and sucking filtrate to obtain the cortex cells and freezing and drying the cortex cells to obtain wool keratin cortex cells. The method is efficient and environmentally friendly.

The invention relates to a cocktail immunohistochemical kit for diagnosing breast cancer, which comprises a peroxidase sealing agent, a combined primary antibody, a combined secondary antibody, a DAB chromogenic substrate, a DAB chromogenic buffer solution, a fast red chromogenic agent, a fast red chromogenic activator, a fast red buffer solution and a hematoxylin restaining solution, wherein the combined primary antibody comprises a mouse anti-human cytokeratin 7 monoclonal antibody, a mouse anti-human P63 monoclonal antibody and a rabbit anti-human MHC monoclonal antibody; the combined secondary antibody comprises an anti-mouse secondary antibody polymer marked by HRP and an anti-rabbit secondary antibody polymer marked by AP. According to the kit, the cocktail combined primary antibody is adopted, the expression conditions of three antigen targets CK7, P63 and MHC can be detected on the same tissue slice at the same time, the use amount of samples is reduced, the difference caused by different slices is avoided, the problem of poor dyeing repeatability of tiny tissue samples is solved, normal tissue and cancer tissue can be visually and clearly distinguished, a more comprehensive and reliable diagnosis basis is provided for pathologists, and the diagnosis accuracy is improved.