3676 results about "In vitro" patented technology

The invention discloses a method for in vitro slow release performance evaluation of a slow and controlled release preparation based on the overflow principle. A container with an upper outlet and a lower outlet is adopted to serve as a release tank, a to-be-detected medicine and a dissolution medium are placed in the release tank, a stirring device is adopted in the release tank to perform stirring, and the dissolution medium is added from the lower outlet of the release tank through a peristaltic pump; the dissolution medium dissolved with the medicine is taken away through the overflow from the upper outlet so as to offset medicine absorption or metabolism, and the slow and controlled release speed of the medicine, the total medicine overflow quantity and the total release amount are calculated by measuring the concentration of the medicine contained in the dissolution medium flowing out, the volume of the release tank and the overflow quantity. According to the method, the releasing degree of the slow and controlled release medicine can be evaluated accurately, the effective medicine release time of the slow and controlled release preparation is evaluated, and slow release performances of the slow and controlled release preparation are evaluated comprehensively.

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

The invention belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody for specific recognition of type II grass carp reovirus VP4 protein, an eukaryotic expression method and application. According to the invention, a target gene S6 is transfected to an HEK293 cell for expression and purification, a New Zealand white rabbit is immunized in cooperation with a Freund's adjuvant, and the rabbit-derived recombinant monoclonal antibody capable of specifically recognizing the GCRV-II VP4 protein is obtained by using ELISA, single B cell screening and eukaryotic expression technologies. The antibody has strong specificity, provides support for further establishment of a specific II-type grass carp reovirus diagnosis technology, and has great application value in development of GCRV-II related scientific research.

This invention provides an in vitro cell-free protein synthesis system, its reagent kit, and its applications, belonging to the field of protein synthesis technology. This invention improves an in vitro cell-free protein synthesis system based on Kluyveromyces lactis by adding exogenous L-arabinose, significantly enhancing the system's protein synthesis capacity. Furthermore, it provides a more efficient and higher-throughput in vitro protein synthesis reagent kit and a method for synthesizing exogenous proteins. The improvements provided by this invention can be achieved without molecular modification, are simple and convenient to operate, and save costs.

The invention provides a PET tracer precursor compound as well as a preparation method and application thereof, and also discloses a < 68 > Ga-labeled PET tracer, the PET tracer precursor compound is simple in preparation method, short in preparation time and high in radiochemical yield, and has the advantages of good in-vivo and in-vitro stability, high specificity, high intake and the like; the tumor or inflammation part can be accurately and stably detected and positioned; the Ga-68 labeled targeted PARP-1 radioactive probe synthesized on the basis of the compound Olaparib can realize qualitative diagnosis of PARP-1 in tumors, further realizes non-invasive accurate quantification of PARP-1 in tumors, provides a reliable basis in stages of clinical staging, treatment decision, curative effect monitoring and the like, and finally realizes accurate diagnosis and treatment.

The invention discloses a biological in-vitro sample aging quality control detection method, and relates to the technical field of biological samples, and the method specifically comprises the following steps: screening metabolic markers based on biological physiological characteristics, and constructing an in-vitro quality attenuation model; collecting and preprocessing sample data; detecting the concentration of the metabolic marker through detection equipment; inputting marker concentration data into the model for dynamic comparison; and outputting aging classification, and guiding experiment scheduling operation. According to the method, effective use time efficiency is adopted for rapid judgment, the problem that experimental data is influenced by metabolic degradation of a sample after in-vitro is solved, time efficiency grading can be completed in a short time through multi-marker dynamic comparison and a machine learning model, the method is remarkably superior to a traditional method, and by combining a micro-fluidic detection device and an LSTM model, the detection accuracy is greatly improved. According to the method, the samples can be scheduled to proper experiment operation according to the effective time limit of the samples, waste of the samples is avoided, and after failure samples are detected, a user can be reminded to collect the samples again, so that the experiment process is prevented from being delayed.

The invention relates to an anti-CD3 nano antibody, an anti-CD38 antibody and a bispecific antibody containing the anti-CD3 nano antibody and the anti-CD38 antibody. According to the invention, an anti-CD3 nano antibody and an anti-CD38 monoclonal antibody are excavated and prepared, humanized design is carried out to obtain antibodies with high affinity and specificity, and different configurations of bispecific T cell conjugation antibodies targeting CD3 and CD38 are further designed, so that the CD3 and CD38 dual-specificity T cell conjugation antibodies are obtained. The antigen binding activity, the T cell-mediated tumor cell killing effect and the T cell proliferation and activation function of the antibody are systematically evaluated in vitro, and experimental results show that the T cell conjugation antibody can effectively recruit and activate T cells and has a remarkable killing effect on CD38 positive tumor cells, and it is further verified that the antibody has remarkable in-vivo anti-tumor activity and has a good application prospect. Therefore, the polypeptide has high affinity, good pharmacological characteristics and development potential.

The invention discloses bifidobacterium breve and application thereof in relieving Alzheimer's disease, and belongs to the technical field of probiotics. The preparation method comprises the following steps: separating 18 bifidobacterium strains from feces of children aged 0-3 years old, analyzing and identifying the 18 bifidobacterium strains including bifidobacterium longum, bifidobacterium breve and bifidobacterium animalis, and screening out the bifidobacterium breve LE4 with potential Alzheimer's disease relieving capacity through in-vitro oxidation resistance and LPS (Lipopolysaccharide) removal capacity. The method comprises the steps of behavioral experiments (nesting experiments, open field experiments and water maze experiments), histopathologic analysis, molecular biological detection and the like. The multi-target AD intervention effects of improving the cognitive function, reducing A beta deposition and tau protein phosphorylation, relieving neuroinflammation, enhancing synaptic plasticity, repairing intestinal barriers, adjusting intestinal flora and metabolites and the like of the LE4 strain are comprehensively evaluated, and a new strategy is provided for promoting AD micro-ecological treatment.

The invention discloses a poultry multi-stage feed net energy in-vitro prediction method and device based on factorial screening, and relates to the technical field of animal husbandry. The method comprises the following steps: collecting and screening poultry feed raw material samples; analyzing the factors of the raw material components influencing the net energy value by adopting a factorization method to obtain a key component set; based on poultry feed raw material samples, determining the content of key components of each feed raw material and net energy values of the poultry feed raw material samples in different growth stages of poultry; based on the measured content of the key components of each feed raw material and the net energy values of the poultry feed raw material samples in different growth stages of poultry, performing principal component analysis to obtain predictive factors; constructing target net energy prediction equations of different growth stages of the poultry by using a linear regression analysis method based on the prediction factors; and performing net energy prediction on the poultry feed raw materials to be detected based on the target net energy prediction models of different growth stages of the poultry to obtain predicted net energy values of different growth stages of the poultry.

The present invention, among other things, provides technologies for detecting and / or quantifying nucleic acids in cells, tissues, organs, or organisms. Pre-designed barcodes are associated with specific molecular targets through sequential hybridization experiments. A pseudo-color based barcoding scheme is described that overcomes the limitations in the previous generation of the technology. The current method can be applied to both in vitro and in situ analysis.

The invention relates to a low-molecular-weight chondroitin sulfate composition as well as a preparation method and application thereof, and aims to solve the problems that a chondroitin sulfate product obtained by a single degradation mode is limited in function and difficult to synergistically regulate intestinal barriers. The preparation method comprises the following steps: firstly, preparing LMCSO through oxidative degradation; secondly, preparing LMCSH through hydrothermal degradation; and finally, the LMCSD is prepared through deamination degradation. Finally, the obtained LMCSO, LMCSH and LMCSD are compounded according to the weight ratio of 5: 3: 2, and the low-molecular-weight chondroitin sulfate composition LMCSs-C is obtained. The composition LMCSs-C prepared by the invention can synergistically improve the intestinal barrier function through multiple action mechanisms, not only shows an obvious intestinal barrier repair effect in vitro and animal models, but also has good biocompatibility and safety, and shows a wide application prospect in intestinal health maintenance and related disease intervention.

The invention discloses a method for transdifferentiating tail fin cells of erythroculter ilishaeformis into fat and muscle cells and application, the method is based on 5-Azacytidine (azacytidine) to promote mesenchymal stem cells to be differentiated into muscle cells, and other small molecule compounds are added on the basis of adding the 5-Azacytidine to transdifferentiate the tail fin cells of erythroculter ilishaeformis into muscles. The method is realized through an in-vitro cell reprogramming technology, fish individuals do not need to be sacrificed, and the destructive influence of traditional muscle or fat sampling on fish bodies is avoided; a single-step induction system does not need multi-factor sequential treatment, a high-purity target cell population can be obtained only in 5-7 days at least, the standardization degree of the operation process is high, the stability among batches is excellent, specific cells can be continuously and stably produced, and a safe and reliable technical basis is provided for sustainable cell culture fish meat production.

The invention belongs to the technical field of in-vitro cultivation of calculus bovis, and particularly relates to a method for in-vitro cultivation of calculus bovis. According to the invention, firstly, the escherichia coli is subjected to bile domestication and ray induced mutation, a high-activity oxgall-resistant bezoar production engineering bacterium E.coli-NH01 is obtained through screening, a method for in-vitro cultivation of bezoar by using the engineering bacterium E.coli-NH01 is provided, the prepared bezoar meets the pharmacopoeia standard, the production efficiency of in-vitro cultivation of bezoar is improved, and the production cost is reduced. And the production cost is reduced, the overall forming period is shortened, and the key that precious traditional Chinese medicinal materials are popular can be achieved.

The invention relates to a polypeptide NbG7-4 capable of identifying and neutralizing SFTSV, which comprises three complementary determining regions CDR1-3, and the sequences of the three complementary determining regions CDR1-3 are respectively shown as SEQ ID NO: 1-3. Nanometer antibody drug development and diagnostic kit research and development are carried out on SFTSV which is high in fatality rate but lacks effective vaccines and specific antiviral drugs, the nanometer antibody VHH which is specifically combined with GN is screened through the platform technology of preparing GN protein, inactivating viruses, immunizing alpaca, displaying nanometer monoclonal antibodies through a phage library and the like, the CDR sequence of the nanometer antibody VHH is identified, and the SFTSV with the high fatality rate is obtained. A humanized antibody NbG7-4 is constructed; meanwhile, the curative effect of the NbG7-4 in treating the SFTSV infection is evaluated at the in-vitro cell level. The invention provides a potential nano antibody new drug for clinical treatment of SFTSV, and also provides a corresponding detection kit for diagnosis of SFTSV.

The present invention belongs to the field of virus detection technology, and specifically relates to monoclonal antibodies with neutralizing activity against adenovirus type 5 and their uses. The monoclonal antibody 8B9 of the present invention can specifically recognize and neutralize adenovirus type 5, and the amino acid sequences of the heavy and light chain variable regions and their complementarity determining regions of the monoclonal antibody 8B9 are provided. The present invention also provides an in vitro neutralization activity assessment model based on HEK-293A cells. At a concentration as low as 3.3 μg / mL, the titer of the neutralizing activity against the virus stock solution after 6000-fold dilution is 7×10 4 TCID 50 / mL of virus has close to 100% neutralizing activity. After further humanization, it is expected to be developed into a neutralizing antibody drug for the treatment of diseases such as severe pneumonia caused by HADV-5 infection, filling the current lack of effective treatment options in this field and possessing significant scientific research value and clinical application prospects.

The present application relates to a virus of reverse genetic operation construction of NADC34-like PRRSV2. Specifically, the full gene sequence of NADC34-like PRRSV2 BJ1805-2 isolated strain is segmented and connected to the vector by using PCR amplification, enzyme digestion and other methods with pACY177 plasmid as the carrier, and the BJ1805-2 full-length infectious clone recombinant plasmid (rBJ1805-2) is obtained. The infectious clone virus rBJ1805-2 is rescued in vitro, and the first NADC34-like PRRSV2 strain infectious clone platform in China is successfully built. The present application also relates to a modification method for constructing NADC34-like PRRSV2 strain which can adapt to Marc-145 cell passage culture. The infectious clone virus constructed by the present application has good in vitro and in vivo proliferation efficiency, and can cause specific viremia after inoculation in pigs; it has good safety, does not cause fever after inoculation in pigs, and will not cause piglet death. The modified strain can be used as a candidate strain for developing the first NADC34-like PRRSV2 specific vaccine in China, and is conducive to the prevention and control of PRRSV epidemic in China.

The invention relates to an in vitro method for classifying a subject afflicted with a cancer as suffering from a cancer with (at risk of) a metabolic reprograming towards fatty acids oxidation including a step of assaying the activation of RelB in a tumor sample form said cancer. The inventor indeed identified the pivotal role of RelB in energy metabolism and more particularly mitochondrial respiration and fatty acid oxidation. Accordingly, the invention also relates to inhibitors of RelB activity or expression, as well as of lipid metabolism for use in the treatment of cancers showing an activated RelB.

Provided are variant Cas12f4 polypeptides, fusion proteins containing variant Cas12f4 polypeptides, nucleic acids encoding variant Cas12f4 polypeptides and fusion proteins, compositions and methods related to those variant Cas12f4 polypeptides and fusion proteins, and modified host cells comprising the variant Cas12f4 polypeptides and / or encoding nucleic acids. The variant Cas12f4 polypeptides disclosed herein exhibit improved functionality, stability, and / or other desirable properties and may be used in combination with guide Cas12f4 RNAs that can be used in a variety of applications ex vivo, in vitro, and in vivo.

The invention discloses clostridium butyricum as well as a culture method and application thereof, belongs to the field of feed additives, and aims to solve the problems that a controllable fattening period is shortened and the efficacy of the clostridium butyricum cannot be completely exerted due to the fact that a wide range of clostridium butyricum needs a relatively long adaptation period for an endogenous intestinal environment of an animal. The clostridium butyricum from the North Naa Biotechnology Co., Ltd. Is cultured by simulating an animal endogenous environment in vitro, and after the clostridium butyricum which completely adapts to animal endogenous is obtained, the screened animal endogenous clostridium butyricum is coated by adopting a coating technology, so that the clostridium butyricum is not damaged by a strong acid environment in an animal stomach bag; the clostridium butyricum strain has the advantages that the clostridium butyricum strain can be used for preparing animal endogenous clostridium butyricum, a large quantity of animal endogenous clostridium butyricum can reach intestinal tracts and be released, and the animal endogenous clostridium butyricum can quickly adapt to animal intestinal environment due to higher adaptability of the strains, and can quickly form efficient colonization and functional expression under the condition of almost no adaptation period. The feed additive can be used as a biological feed additive.

The invention discloses an SaRNA vaccine for echinococcosis as well as a preparation method and application of the SaRNA vaccine. The preparation method of the SaRNA vaccine comprises the following steps: carrying out codon optimization on a modified target antigen protein through a genetic engineering technology, then assembling the modified target antigen protein with a self-replicating protein sequence, 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a plasmid, and carrying out purification to obtain the SaRNA vaccine. The preparation method comprises the following steps: constructing recombinant plasmids, sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmids to prepare SaRNA molecules, and finally wrapping the SaRNA molecules in lipid nanoparticles to form the SaRNA vaccine for the echinococcosis. Experiments prove that the SaRNA vaccine can activate humoral immunity and cellular immunity of mice at the same time, high-level EG95 specific antibodies and cytokines can be generated through low-dose immunity, and the SaRNA vaccine has wide application prospects in the aspect of preventing and / or treating the echinococcosis.

The invention relates to a method for inhibiting in-vitro infection of a porcine epidemic diarrhea virus by using JPH203 and application. The JPH203 can be used for preparing an antiviral drug for inhibiting infection of the porcine epidemic diarrhea virus. JPH203 with the concentration of 5 [mu] M is added in the process that Vero cells are infected with PEDV, compared with a control group without JPH203, the virus infection condition, the protein expression quantity, the RNA content and the virus titer of the PEDV in a treatment group with JPH203 are all remarkably reduced, and it is indicated that JPH203 can be used for inhibiting in-vitro infection of the porcine epidemic diarrhea virus.

Provided herein is a method for efficiently capping RNA in vitro. In some embodiments the capping reaction may be done at high temperature using Vaccinia capping enzyme or a variant thereof. In other embodiments, the capping reactions may comprise a capping enzyme from a large virus of amoeba, e.g., Faustovirus, mimivirus or moumouvirus, or a variant thereof. Compositions and kits for practicing the method are also provided.

The invention relates to the field of preparation and application of biological peptides, in particular to a swelling hippocampal peptide and application thereof in preparation of uric acid reducing and gout resisting products. The amino acid sequence is shown as SEQ ID NO.1-5. The uric acid reducing effect is verified by adopting an in-vitro XOD inhibition experiment and a cell experiment; furthermore, the uric acid reducing and gout resisting activity of the compound is verified through a hyperuricemia animal model, and a theoretical basis is provided for high-valued utilization of the belly-swelling hippocampus and development of uric acid reducing and gout resisting products.

The invention relates to euphausia superba peptide with uric acid reducing activity and application thereof. The euphausia superba peptide is a short peptide obtained by extraction, enzymolysis, separation and purification from degreased euphausia superba powder, and the amino acid sequence of the euphausia superba peptide is Ser-Ala-Gly-Lys-Thr. Researches show that the peptide can reduce the generation of uric acid by inhibiting the activity of xanthine oxidase and promote the excretion of uric acid. In-vitro cell experiments prove that under the concentration of 10 mu M, the inhibition rate of the peptide on xanthine oxidase reaches 23.1%; animal experiments show that the blood uric acid level of mice in a high-dose group (50mg / kg) is reduced by about 40% compared with that in a model group. The invention also develops a health-care beverage and a medicinal preparation which take the peptide as an active component, and provides a new choice for preventing and treating hyperuricemia and gout.

The invention discloses a lipid compound capable of being used for gene delivery, a preparation method of the lipid compound and application of the lipid compound in gene delivery. Also disclosed herein is a lipid nanoparticle comprising the lipid compound, a gene delivery composition comprising the lipid compound or the lipid nanoparticle. The lipid compounds, lipid nanoparticles and delivery systems herein enable efficient complexation, protection, intracellular and targeted delivery and release of biomolecules, such as oligonucleotides and nucleic acids, in vitro and in vivo tissues and organs.

The application discloses a method for identifying a protospacer adjacent motif, and relates to the technical field of biology. The method uses the nucleic acid cutting activity of CRISP-Cas protein, detects the specificity of CRISP-Cas protein to different PAM sequences in vitro by using a DNA array with 256 PAM sequences, and can clearly show the recognition preference of CRISP-Cas protein to each PAM sequence. Meanwhile, the method is simple in steps, easy to operate, does not need sequencing, and short in detection time, and can be used as a general detection method for CRISP-Cas protein.

The invention discloses application of bifidobacterium adolescentis CCFM1302 in promoting synthesis of host collagen, and belongs to the technical field of microorganisms and medicines. The metagen of the bifidobacterium adolescentis CCFM1302 disclosed by the invention has a good effect of regulating and controlling synthesis and degradation of collagen, the moisture and elasticity of the skin are improved, the antioxidant capacity of the skin is improved, the inflammation level of an organism is reduced, and the stable state of collagen is maintained. The method specifically comprises the following steps: in vitro promoting the enzyme activity of HSF cell LH1 by a thallus lysate, inhibiting the enzyme activity of MMP-3, increasing the content of COL I and COL III and the enzyme activity of LH1, P4H and LOX from the outside of a fermented supernatant, and inhibiting the enzyme activity of MMP-3; the inactivated thalli are orally taken to increase the collagen content and the enzyme activity of LH1 and inhibit the enzyme activity of MMP-1 and MMP-3; the fermentation supernatant is orally taken to inhibit enzyme activity of MMP-3 and MMP-1 and degradation of collagen, and the content of TGF-beta is increased.

The invention relates to the technical field of medicines, in particular to a construction method and application of a photothermal conversion platform for treating osteoarthritis by regulating synovial macrophage polarization. According to the present invention, the M2 membrane-structured black phosphorus selenium carbon body (M2-BPSeC) photo-thermal nano-platform is innovatively provided, the triple functions of bionic targeting delivery, precise thermal control intervention and immune microenvironment remodeling are integrated, and the excellent synergistic treatment effect is represented in the in-vitro cell model and the OA animal model. The technology breaks through the technical bottleneck that traditional treatment is insufficient in targeting and single in intervention dimension, precise functional coupling of the physical thermal effect and immunoregulation is achieved for the first time, and a brand new strategy of targeting-temperature control-immunity integration is provided for osteoarthritis treatment. The efficient and controllable preparation process and the remarkable multi-dimensional treatment effect open up a new path for cartilage regeneration and immune balance regulation and control.

The invention belongs to the technical field of assisted reproduction, and particularly relates to a device and a method for gradually adjusting the glucose concentration of an in-vitro embryo culture solution. The device comprises at least two miniature liquid storage bins, wherein each miniature liquid storage bin is filled with a high-concentration glucose solution in advance; the slow release channel is respectively connected with each micro liquid storage bin and the culture solution main container; the culture solution main container is used for accommodating an embryo and a basal culture medium; the control unit is used for sequentially opening the release ports of the miniature liquid storage bins, so that the high-concentration glucose solution sequentially flows into the culture solution main container through the slow release channels. The technical means of staged closed-loop micro sugar control is introduced into the structure and the using method of the device, so that the accurate adjustment of the sugar concentration of the in-vitro embryo culture solution is realized, the requirements of embryos in different development stages can be effectively met, and the embryo culture quality and the subsequent development potential are remarkably improved.

The invention belongs to the technical field of preparation of polypeptides, and particularly relates to a self-assembled polypeptide capable of responding to alkaline phosphatase and degrading Neuropilin-1 as well as a preparation method and application of the self-assembled polypeptide. The self-assembled polypeptide (formula I) provided by the invention has a self-assembly starting unit 'GNNQQNY (SEQ ID NO: 1)', can be self-assembled in vitro, and can form nanofibers after responding to alkaline phosphatase, so that not only can the critical self-assembly concentration of the polypeptide be reduced, but also the binding capacity with target protein can be improved. The material can degrade NRP1 on the surfaces of tumor cells and Treg cells, and has huge potential in promoting immunotherapy of tumors.