256 results about "Functional studies" patented technology

The present invention pertains to the identification of moieties and methods of using the same for preventing tolerance to bronchodilators. More specifically, the present invention pertains to the identification of compositions and methods which are capable of preventing tolerance to beta2-adrenergic agonists. The methods and compositions according to the invention are also useful as analytical tools for functional studies and as combination therapeutic tools.

The invention discloses a simple, convenient and efficient fixed-point gene editing method relating to the fields of transgenic animal preparation, gene functional study and biomedicines. A testis injection method is a sperm-mediated gene transferring method developed by utilizing the sperm capacity of actively combining, transferring and integrating an exogenous DNA. By using the method disclosed by the invention, an animal testis injection method is optimized, a carrier constructed by utilizing a clustered regularly interspaced short palindromic repeat system (CRISPR / Cas9) is integrated with a sperm chromosome through multi-point testis injection or seminiferous tubule microinjection, so that the aims of deleting, replacing and inserting genes are achieved. Then, a transgenic animal descendant is obtained through various approaches such as natural mating and artificial insemination. According to the method, the advantages of a CRISPR / Cas9 gene editing system are sufficiently utilized, and the existing transgenic animal system is combined, so that the complexity of in-vitro cell operation and requirement on expensive precise instruments / equipment are avoided while the gene modification animal obtaining efficiency is greatly increased.

The present invention discloses a method for establishing KI-T2A-luciferase cell line based on CRISPR / Cas9 targeted genome modification technology. A T2A-luciferase reporter gene is integrated in the3<rd> end of the mmp 12 gene in a genome by using CRISPR / Cas9 technology. A knock-in cell line of MMP12-T2A-luciferase is established, the site-specific integration of the source gene in the cell lineon the genome is verified. Meanwhile, a reported transcription factor, STAT3, with activation effect on mpp12-T2A is used to transcribe and active the MMP12-T2A-luciferase cell line. The results showthat the expression level of luciferase in MMP12-T2A-luciferase cell line can accurately and sensitively reflect the expression level of MMP12 protein in the cell line. The establishment of the cellline will contribute to the study of the gene function of mmp12 and the screening of small molecule chemical drugs affecting the expression of mmp12, which provides a new experimental thinking and solution for the migration of cancer cells and related researches thereof.

The invention relates to a method for establishing luciferase knock-in cell line based on CRISPR-targeted genome modification technology. The knock-in cell line of SREBP1- T2A-Luciferase was established by in situ integration of the 3'end of SREBP1 gene into the T2A-Luciferase reporter gene using CRISPR / Cas9 technique, the fixed point of the Chinese and foreign source genes of the cell line on thegenome is verified. The transcriptional activation of the SREBP1-T2A-Lucifasse cell line was performed using the reported transcription factor LXR Alpha, which has an active effect on the SREBP1. Theresults show that the expression level of the Luciferase in the SREBP1-T2A-Lucifasse cell line can accurately and sensitively reflect the expression level of the SREBP1 in the cell line. The establishment of the cell line will help to study the function of the SREBP1 gene and to screen the small molecular chemical drugs affecting the expression of SREBP1, and provide a new experimental thought and solution for lipid metabolism and related research.

The invention discloses a brain function network modeling method for resting state synchronization EEG-fMRI, and relates to the research field of neural signal processing. The brain function network modeling method includes the steps that firstly, EEG signal preprocessing is conducted, a tape limit energy signal is extracted and a regression item is established; secondly, fMRI signal preprocessing is conducted, and a BOLD signal of each brain region is extracted; thirdly, the BOLD signal, extracted through fMRI, of each brain region and the regression item obtained from EEG are subjected to principal component analysis; fourthly, canonical correlation analysis is conducted on principal components of the two signals obtained from the last step; fifthly, modeling is conducted on a brain function network of resting state synchronization EEG-fMRI. By means of the brain function network modeling method, a plurality of the brain function networks can be obtained; the brain function networks and existing research results have high coherence so that the effectiveness of the modeling method can be proved, and thus a new thought and a scheme are provided for the resting state brain function research.

The invention discloses a method for determining influence of exogenous dsRNA on toxicity of ladybugs. The method comprises the following specific steps: uniformly mixing the exogenous dsRNA into a sucrose solution, directly feeding the ladybugs for 2-3 days and then feeding with pea aphids; then detecting and analyzing the expression changes of target genes in the ladybugs, and observing the biological changes of the ladybugs so as to evaluate the toxicity of the exogenous dsRNA to the ladybugs, wherein the ladybugs include harmonia axyridis, coccinella septempunctata or henosepilachna pusillanima. By the method, direct influence of the exogenous dsRNA on the toxicity of the harmonia axyridis, the coccinella septempunctata or the henosepilachna pusillanima or other ladybugs can be effectively determined; in addition, the method is simple, feasible and high in effectiveness and sensitivity, and has a great significance and an application prospect in study on functions of related genesas well as environmental risk assessment on related RNAi transgenic crops.

The invention discloses a method for building a virtual mannequin based on bodily form of actually measured crowd, which comprises: a) due to direct method of measurement, 100-400 human body samples are chosen according to different districts and age periods, and 40-70 unary data are obtained by measuring various parts of human body; b) the data are analyzed, the data of medium bodily form of different districts and age periods are obtained; c) the samples of not less than 200 people are chosen for carrying out 3D untouched type human body measurement; d) 3D model of human body is pretreated, standardization is carried out by using cloud data of human body model, at least 100 mannequins are obtained, the better is 120-180 sections, each section uses 40-70 mannequins, and the better is representation of 66 data points; e) 3D average form of the sample of human body model is calculated, and the 3D virtual mannequin is obtained by revision disposal. According to the method of the invention, mannequin entity satisfying actually measured bodily form can be formed in a short time; and according to different districts and age periods, bases and data are provided for suitability of clothes production, virtual fitting and cutting-out, clothes function study, specification standard and antitype of bodily form of the people in our country. The period for designing mannequin is shortened.

The invention discloses an individual brain function mapping method based on electrocorticogram high-frequency Gamma nerve oscillation and belongs to the field of nerve engineering. The individual brain function mapping method based on electrocorticogram high-frequency Gamma nerve oscillation comprises the steps that collected ECoG data are processed, the correlation synchronization intensity of time-frequency events is calculated, statistical significance testing is conducted and a brain function index is extracted, and individual brain function mapping is conducted. According to the individual brain function mapping method based on electrocorticogram high-frequency Gamma nerve oscillation, through the deep mining of the ECoG data, relevant electroencephalogram data of brain function areas can be analyzed rapidly, accurately, comprehensively and systematically, the brain function index is obtained, individually targeted brain function mapping is achieved, people can better understand brain mechanisms for processing complicated cognition tasks, and great assistance is provided for study on cognitive neuroscience brain functions and fundamental study on clinic neurosciences.

The invention discloses an assistant hypoglycemia healthcare food of mulberry-leaf extract and a preparation method thereof. The method comprises the following steps: 1) placing the following components into a boiling granulator in parts by weight: 2.0-7.0 parts of a mulberry-leaf extract, 1.0-4.0 parts of an astragalus root extract, 0.5-2.5 parts of a solomonseal extract, 0.5-2.0 parts of a kudzuvine root extract, 0.00005-0.0002 part of chromium picolinate, starch and dextrin, starting the boiling granulator to fully and evenly mix raw materials and auxiliary materials, taking water as a wetting agent for spray granulation, stopping spraying, drying for 0.5-1 hour, and then detecting 5-7% of moisture; 2) granulating the prepared particles by a particle granulation machine with a 20-40 mesh sieve; and 3) adding a disintegrating agent-carboxymethyl starch sodium and a lubricating agent-magnesium stearate to the granulated particles in a multidirectional movement mixer, fully and evenly mixing, and finally filling capsules or pressing into tablets. The assistant hypoglycemia healthcare food of mulberry-leaf extract is safe and effective and has small administration dosage; and toxicology and functional studies show the assistant hypoglycemia capsules or the tablets is safe and nontoxic, has obvious effects and assistant hypoglycemia function.

The invention provides a new production process for large-scale synthesis of long-chain RNA drugs. The process is characterized by specifically including the steps of: DNA transcription template design; template preparation and purification; in-vitro transcription; and product purification, purity detection and the like. The production process provided by the invention is especially suitable for synthesis of long-chain RNA drugs with a stable secondary structure. With the characteristics of simple operation and low cost, the process provided by the invention meets the requirements of large-scale production, and the produced long-chain RNA drugs can be used for cell experiments, animal experiments and other preclinical RNA drugs' functional study.

The invention relates to the technical field of biotechnology, in particular to a method for building a special microRNA knock-down mouse model of a heart. The invention provides a transgenic mouse built by using a miRNA sponge technology. Exogenous gene components comprise a myocardium special promoter, a miR-328 sponge and a transcription termination signal. The myocardium miR-328 of the mouse is knocked down by almost 50%, and the mouse model is suitable for selection of antiarrhythmic drugs because the miR-328 is involved in the occurrence of arrhythmia. The method is based on the transgenic mouse, so the model can stably express the sponge to stably knock miRNA down and can inherit simultaneously. The mouse phenotype is stable, thus the mouse model is applicable for study of subsequent functions.

The present invention provides cloning sequencing, analyzing, function research of a biosynthesis gene cluster of an antibiotic-Azinomycin B having antitumor activity produced by streptomyces, and its application. The whole gene cluster includes 34 genes: one repeatedly using I type polyketide synthase gene; two naphthalene ring modification enzyme genes; 8 non-ribosomal polypeptide skeleton synthesis and modification enzyme genes; 11 non-natural amino acid structure unit synthase genes; 1 resistance gene; 3 post modification enzyme genes and 8 genes which functions are not determined. The genetic operation of the biosynthesis gene breaks the synthesis of Azinomycin B; the precursor compound is produced by the heterologous expression of synthesis gene and modification gene of naphthalene ring. The gene of the invention and the protein can be used for searching and finding compound or gene, protein applied in medical, industry or agriculture.

The invention discloses an application method of virtual mannequin. The virtual mannequin consists of at least 100 sections and 40-70 data points which represent all the sections respectively. The file format of the mannequin is physical data defined by CAD software and file of VRML format. The application method comprises that: the native places and birthplaces of father and mother of the applied object and the birthplace of the applied object are confirmed; and the district bodily form of the applied virtual mannequin is confirmed; the age period of the applied object is confirmed; comparing with the physical data of mannequin defined by CAD software and file of VRML format, the data of the bodily form of the applied object is input, and the applied virtual mannequin is confirmed again. Therefore, research precision can be improved greatly in a short time, and according to different districts and age periods, bases are provided for suitability of clothes production, making of specification standard of clothes, research and analysis of clothes function and accurate data measurement of human body.

The invention discloses a monopterus albus gene editing method. The monopterus albus gene editing method comprises the following steps: 1, acquiring an artificially inseminated monopterus albus 1-cell stage embryo by establishing an indoor eel whole artificial propagation technology; 2, establishing a transcription activator-like effector nuclease gene editing vector of a target gene, performing in-vitro transcription to synthesize mRNA, and transferring the mRNA into the monopterus albus 1-cell stage fertilization embryo by adopting a microinjection method; 3, screening to obtain a monopterus albus with a mutant target gene through a three-primer detection method, and determining the mutation type of the target gene through sequencing. According to the method, a micromanipulation technology for a monopterus albus embryo is established for the first time, a precise monopterus albus endogenous gene editing technology is established for the first time, and powerful technological means is provided for performing function research on a monopterus albus gene and genetic improvement of monopterus albus breeding variety.

Methods and oligonucleotide reagents for analyzing individual T cells are disclosed. In particular, the present disclosure provides methods for analyzing individual T cells using high-throughput multiplex amplification and deep sequencing of nucleic acids encoding T cell receptors (TCRs) and various other T cell phenotypic markers. The present disclosure further provides methods of reconstituting TCRs from individual T cells for functional studies, ligand discovery, or screening therapeutics.

The invention discloses applications of staurosporine compounds in preparing medicines for treating cancer, inflammation or AIDS. The compounds are obtained through separation and purification from actinomycetes, and are identified to be staurosporine compounds. The functional research shows that the compounds have relatively high activity for prostate cancer cells, and also have a relatively highinhibiting effect for Brd4 protein, and therefore, the compounds have good application prospects in preparing the medicines for treating tumor, HIV, leukemia and other diseases.

Targeting metabolic enzymes in human cancer Abstract Lung cancer is a devastating disease and a major therapeutic burden with poor prognosis. The functional heterogeneity of lung cancer (different tumor formation ability in bulk of tumor) is highly related with clinical chemoresistance and relapse. Here we find that, glycine dehydrogenase (GLDC), one of the metabolic enzyme involved in glycine metabolism, is overexpressed in various subtypes of human lung cancer and possibly several other types of cancers. GLDC was found to be highly expressed in tumor-initiating subpopulation of human lung cancer cells compared with non-tumorigenic subpopulation. By array studies we showed that normal lung cells express low levels of GLDC compared to xenograft and primary tumor. Functional studies showed that RNAi inhibition of GLDC inhibits significantly the clonal growth of tumor-initiating cells in vitro and tumor formation in immunodeficient mice. Overexpression of GLDC in non-tumorigenic subpopulation convert the cells to become tumorigenic. Furthermore, over-expression of GLDC in NIH / 3T3 cells and human primary lung fibroblasts can transform these cells, displaying anchorage-independent growth in soft agar and tumor-forming in mice. Not only is GLDC is expressed human lung cancer, it is also up-regulated in other types of cancer, such as colon cancer. RNAi knockdown of GLDC in colon cancer cell line, CACO-2 cells, can also inhibit the tumor formation in mice. Thus GLDC maybe a new metabolic target for treatment of lung cancer, and other cancers.

The invention relates to a cotton GbSTK gene and encoding protein thereof. Sea island cotton Pima90-53 is taken as a material, a cDNA-AFLP technology is used for screening to obtain differential fragments related to verticillium wilt resistance, and then, a reverse transcription-polymerase chain reaction (RT-PCR) and a RACE (rapid-amplification of cDNA ends) technology are used for amplifying to obtain a resistance gene of cotton verticillium wilt, namely a serine / threonine protein kinase (GbSTK) gene. The functional studies of the gene and the encoding protein thereof show that the GbSTK gene has certain effect on resisting verticillium wilt.

The invention discloses cloning, sequencing, analysis, function study and application of a biosynthesis gene cluster of a novel anti-microbial and anti-tumor compound WSS2277 in the field of biotechnologies. The gene cluster comprises nucleotide sequences of twenty-one genes or complementary sequences which are formed by the sequences 1, wherein the fourteen genes including siaA, siaB1, siaB2, siaC, siaD, siaE, siaF, siaG, siaH, siaI, siaJ, siaK, siaL and siaM are responsible for the synthesis of a carbon skeleton, and the seven genes incluidng siaN, siaO, siaP, siaQ, siaR, siaS and siaT are responsible for regulating biosynthesis. The information of the genes which are related to the biosynthesis of the WSS2277 is provided by the invention, and a foundation is provided for the study and the genetic modification of the biosynthesis of the WSS2277. A novel structural analogue can be generated through the modification of carbon skeleton synthesis genes in the WSS2277, and the yield of the WSS2277 or a derivative thereof can be provided through the interruption or the high replication of regulator genes in the WSS2277. The genes and proteins, which are provided by the invention, can also be used for searching and discovering compounds or genes and proteins, which can be applied to the fields of medicine, industry or agriculture.

The invention discloses a method for expression of fatty acid desaturase by an acellular protein synthesis system, and belongs to the enzyme engineering field. The method uses the wheat germ acellular protein synthesis system for cloning and expression of omega 3 desaturase gene (FADS15) from mortierella alpine ATCC32222 (American type culture collection32222) without the need for preparation of mRNA, the expression amount is high, subsequent purification steps are simple, and the method lays the foundation for the next step of research of membrane protein crystal structures and functions.

The present inventors developed hepatitis C virus 1a / 2a and 1b / 2a intergenotypic recombinants in which the JFH1 structural genes (Core, E1 and E2), p7 and NS2 were replaced by the corresponding genes of the genotype Ia reference strain H77C or TN or the corresponding genes of the genotype Ib reference strain J4. Sequence analysis of recovered 1a / 2a and 1b / 2a recombinants from 2 serial passages and subsequent reverse genetic studies revealed adaptive mutations in e.g. p7, NS2 and / or NS3. In addition, the inventors demonstrate the possibility of using adaptive mutations identified for one HCV isolate in generating efficient cell culture systems for other isolates by transfer of mutations across isolates, subtypes or major genotypes. Furthermore neutralization studies showed that viruses of e.g. genotype 1 were efficiently neutralized by genotype Ia, 4a and 5a serum, an effect that could be utilized e.g. in vaccine development and immunological prophylaxis. The inventors in addition demonstrate the use of the developed systems for screening of antiviral substances in vitro and functional studies of the virus, e.g. identification of receptors required for HCV entry

The invention relates to the cloning, sequencing, analyzing, functional study of biological synthetic gene cluster of erythromycin produced by streptomycete in tetrahydroisoquinoline alkaloid family which has the antineoplastic activity and application thereof. The whole gene cluster contains 30 genes: 3 nonribosome polypeptide synthetase genes, 4 antecedent molecule synthetic genes of 3-hydroxy-5-methyl-O-methyl-tyrosine, 2 peptid ring skeleton synthetic genes, 7 erythromycin modifying genes, 5 genes related to S-adenosine methionine synthesis, 3 regulation genes, 2 resistance genes and 4 genes with uncertain function. Genetic manipulation of the genes can block the synthesis of erythromycin, change the output and get analogue of erythromycin. The gene cluster can be used for gene engineering, protein expression, enzymic catalytic reaction, etc. of tetrahydroisoquinoline alkaloid family and finding out compounds or genes being suitable for medicine, industry or agriculture.

The invention discloses a diaphragma juglandis acidic polysaccharose and a preparation and an application thereof. The diaphragma juglandis acidic polysaccharose is prepared from polysaccharide with the weight percent being 99% or above, wherein the polysaccharide comprises galacturonic acid, glucose, galactose and arabinose at the mass ratio of (66.30-72.50) to (13.50-18.60) to (6.20-11.40) to (2.80-4.40); the preparation method of the diaphragma juglandis acidic polysaccharose comprises the following steps: extracting diaphragma juglandis powder crude polysaccharide by water extraction and alcohol precipitation, removing protein by employing an enzyme-Sevage combined technique, dialyzing and purifying by anion exchange chromatography and gel filtration chromatography; and carrying out vacuum freeze-drying, separating and purifying to obtain the diaphragma juglandis acidic polysaccharose. The purified polysaccharide is subjected to component analysis, structural identification and immune function research, so that the diaphragma juglandis acidic polysaccharose has relatively high anti-oxidation activity, can be used as an antioxidant or applied to preparation of the antioxidant, and can be widely applied to cosmetics, food supplements, animal feed additives, medicines and the like.

DNA variants may be classified according to a rules-based scoring system into five categories that include pathogenic, likely pathogenic, variant of unknown significance, likely benign, and benign. Scores may be associated with variants in a framework that weighs evidence from prediction tools, population frequency, co-occurrence, segregation, and functional studies. A standardized scoring system for assessing pathogenicity may provide reliable, consistent pathogenicity scores for DNA variants encountered in a clinical laboratory setting.

The invention relates to a clone, sequencing, analysis and function study of biosynthesis gene cluster of antibiotics-Tectrocarcin A which has anti-tumor activity and is generated by Micromonospora chalcea NRRL11289 and the application thereof. The whole gene cluster contains 36 genes: 5 non-reused I type polyketone synthetase genes; 4 special three-carbon biosynthesis genes; 11 desoxy sugar biosynthesis genes; 9 post modifying genes; 2 regulator genes; 1 resistance gene; and 4 genes with uncertain function. Due to the genetic manipulation of biosynthesis genes, the biosynthesis of Tectrocarcin A can be prevented. The genes and the proteins thereof which are provided by the invention can also be used for searching and discovering the compounds, genes or proteins that can be applies in medicine, industry or agriculture.

The invention discloses a self-guidance diffused light tomography method for near-infrared brain function research. The self-guidance diffused light tomography method comprises the following steps: firstly, obtaining the light intensity of a detection point position corresponding to each source point; obtaining a two-dimensional topological image of which a detection area absorption coefficient is changed for the light intensity of the detection point position according to MLBL-OT, and performing image segmentation to achieve the effective positioning of an absorption coefficient change area, and generating an area positioning template matrix K; solving according to a DOT reconstruction method by adopting a Newton-Raphson iteration method, calculating a Jacobi matrix J of a node to obtain an OT-DOT reconstruction equation; calculating to obtain absorption coefficient change in predetermined detection, and drawing an OT-DOT reconstruction image. According to a near-infrared brain function self-guidance imaging method, the DOT inverse problem under-determinedness can be improved in a near-infrared optical modal measuring mode according to an MLBL-OT positioning brain function change area, so that the reconstruction speed is increased.

The invention discloses a culturing method of osteoclasts utilizing mesenchymal stem cells and cytikines, mainly utilizing the mesenchymal stem cells from mouse bone substance and the mouse splenic mononuclear cells and then adding nuclear factor kB receptor activator ligand and macrophage-colony to stimulate the factors. The invention has the advantages: the invention has the advantages of simpleoperation, convenience and practicality, capability of getting osteoclasts at the earliest time, and saving the culture time; the invention gets a large quantity of osteoclasts, solves the problem that osteoclasts are difficult to prepare in large scale, and can meet the need of a general cell biology experiment; the osteoclasts gotten through the invention has high maturity and strong bone substance absorptive capability, which is favorable for the function study; the cell factors needed by the invention is few, which saves expenditure.

The invention relates to the cloning, sequencing, analysis and function research of a biosynthetic gene cluster with Thiostrepton, an antibiotic that resists Gram-positive bacteria and is generated by streptomyces and an application thereof. The whole gene cluster contains 22 genes in total; wherein, 10 genes are biosynthesized into related genes with Thiostrepton macrocycle, 8 genes are biosynthesized into related genes with a side chain, and the functions of the other 4 genes are unknown. The synthesis of the Thiostrepton can be blocked through the genetic operation on the biosynthesized genes. The genes and proteins thereof of the invention can also be used for seeking and discovering compounds or genes or proteins which can be used in medicine, industry or agriculture.

The invention relates to a construction of cucumis interspecific introgression line populations, and a method for applying the same in cucumber breeding, and belongs to the breeding field of biotechnology. A first interspecific introgression line populations of the cucumis is constructed by interspecific cross-backcross-self-cross of wild sour cucumber and cucumber and SSR molecular marker-assisted selection, wherein the interspecific introgression line populations are composed of 86 strains. The introgression line populations comprise introgressed segments of the wild sour cucumber covering 54% of whole genome of the cucumber; and three resources "IL2", "IL24" and "IL49" with relatively strong disease resistance are screened out through inoculation in field; the three resources are crossed with excellent cultivated cucumber respectively to obtain a series of cucumber novel spices with relatively strong disease resistance. The cucumber / sour cucumber interspecific introgression line populations not only broadens hereditary basis of the cucumber but also provides novel genetic resources for development and utilization of excellent genes of the wild sour cucumber, and lays a solid foundation for positioning important genes or genetic loci and functional study of the cucumber.

The invention relates to cloning, sequencing, analysis and functional study of a biosynthetic gene cluster of a natural product FR901464 which has anti-tumor activity and is generated from pseudomonas as well as the application thereof. The whole gene cluster contains 20 genes: five polyketide synthase genes, one hybrid polyketide / non-ribosomal polypeptide synthase gene, one non-ribosomal polypeptide synthase gene, three independent acyltransferase genes, four genes related to polyketide backbone alkylation, four post-modification genes and two control-related genes. The genetic operation of the biosynthetic gene can block the biosynthesis of FR901464. The provided gene and protein thereof can be used for genetic engineering, protein expression, enzyme catalysis reaction, and the like of the compound and can also be used for searching and discovering compounds that can be used in medicines, industry or agriculture or genes and proteins thereof.