740 results about "Molecular mechanism" patented technology

A system and method for determining individualized medical intervention for a particular disease state, and especially for cancers, that includes the molecular profiling of a biological sample from the patient, determining whether any molecular findings including one or more genes, one or more gene expressed proteins, one or more molecular mechanisms, and / or combinations of such exhibit a change in expression compared to a reference, and identifying a non-specific disease therapy or agent capable of interacting with the genes, gene expressed proteins, molecular mechanisms, or combinations of such molecular findings that exhibited a change in expression.

The invention relates to the technical field of gene knockout, and particularly discloses a method for breeding a tcf25 gene deletion type zebra fish through gene knockout. The method comprises the steps of through a CRISPR / Cas9 gene editing technology, designing an appropriate targeting gene locus on a tcf25 gene of the zebra fish, synthesizing in vitro to obtain specific sgRNA and Cas9-mRNA, microinjecting into a cell of the zebra fish, and after culturing an embryo for 60h, selecting the embryo for carrying out genotyping, so that the effectiveness of the selected locus is verified. The method provided by the invention is lower in off-target rate, removes the tcf25 gene through interference, is beneficial to further revealing the whole process of cardiac morphogenesis and a molecular mechanism regulating the process through researching functions through a genetics means, and is of great importance on understanding a cardiac disease pathology and researching and developing a new therapeutic schedule medically.

The invention searches the novel micromolecule inhibitor pyrazolo (1, 5-a) miazines compounds of cyclin-dependent kinase CDK9 (cyclin-dependent kinase) through the virtual screening of a computer, biometrically measures activity thereof, and validates interaction mechanism. The invention specifically comprises the following steps: the three-dimensional crystal conformation of the cyclin-dependent kinase family member CDK9 is obtained in a way of homology modeling; and micromolecule three-dimensional database is screened with DOCK (molecular docking). The invention uses a MTT tumor cell growth inhibition test to biometrically measures the activity of the selected compounds, researches the selected compounds pyrazolo (1, 5-a) miazines with high activety in a way of molecular mechanism, validates the inhibiting effect of the compounds to the activity of CDK9 kinase, and clarifies the interaction mechanism of the compounds for inhibiting the external activity and the molecule of various malignancies such as lung cancer, osteosarcoma, oophoroma, cervical carcinoma, breast cancer, etc.

The invention provides a humanized PD-L1 tumor cell line MC-38-hPD-L1, a builtanimal tumor model with the same and a method for constructing the humanized PD-L1 tumor cell line. The method particularly includes knocking out animal-origin PD-L1 by the aid of CRISPR-CAS9; carrying out amplification and cultivation to obtain knocked-out cell banks; extracting DNA (deoxyribonucleic acid) and carrying out PCR (polymerase chain reaction) amplification; recycling and cloning amplification products; carrying out over-expression on human-origin PD-L1 in MC-38 cell lines of mPD-L1 KO by the aid of lentivirus systems; packaging lentivirus and screening Puromycin to obtain the humanized MC-38 cell line of PD-L1. The humanized PD-L1 tumor cell line, the animal tumor model and the method have the advantages that as shown by results, high killing efficiency and multiplication capacity are obviously presented by tumor infiltration CD8 T lymphocytes after antibody treatment is carried out, tumor infiltration Treg cells can be obviously inhibited after antibody treatment is carried out, and accordingly the method is proved to be effective and feasible from the aspect of molecular mechanisms.

The invention reveals that endogenous cholesterol metabolic product cholestane-3 beta, 5 alpha, 6 beta-triol (YC-5) has protection function for neuronal damage caused by cerebral ischemia, ischemia ofspinal cord, epileptic seizure and convulsion and relates to an application of YC-5 in preparation of neuronal protection medicine. The YC-5 can be prepared from cholesterol through two steps. The primitive cell culture in vitro and spinal cord ischemia animal model proves that YC-5 has protection function for neuronal damage caused by cerebral ischemia, ischemia of spinal cord, epileptic seizureand convulsion. YC-5 in effective dose has no toxic and side reactions. YC-5 is an effective neural protectant which aims to multiple molecular mechanism and cures cerebral ischemia damage, spinal cord ischemia damage, epileptic seizure and convulsion.

The invention discloses a goat TLR4 gene knock-out vector and a construction method thereof. The construction method comprises the following steps: firstly, designing an sgRNA fragment of the TLR4 gene by adopting a CRISPR / cas9 system, synthesizing an sgRNA nucleotide sequence, constructing and simultaneously expressing the sgRNA and plasmid PYSY-sgRNA of Cas9 D10A, connecting and transforming to an Escherichia coli DH5 alpha competent cell, and verifying the transformant; and judging and proving by enzyme digestion and sequencing that the TLR4 gene knock-out vector is constructed correctly. The invention adopts the CRISPR / cas9 for constructing the vector, and provides a theoretical basis for subsequently acquiring a goat TLR4 gene deletion type alveolar epithelial cell system, and studying the immune response molecular mechanism of mycoplasma pneumonia infection.

The present invention provides compounds of formula I: formula (I) wherein X1-X4 and R1-R12 have any of the values defined in the specification, as well as salts and prodrugs thereof, which inhibit major molecular mechanisms associated with bacterial cell division and proliferation so as to be useful for the treatment and / or prevention of bacterial infections. The invention also provides compositions comprising these compounds as well as methods for using these compounds to inhibit bacterial cell division and proliferation and to treat bacterial infections.

The invention discloses a goat CDK2 (Cyclin-dependent kinases 2) gene knockout vector and a construction method thereof. A CRISPR / Cas9 system is adopted; an SgRNA segment of a CDK2 gene is designed at first and an SgRNA nucleotide sequence is synthesized; a plasmid PYSY-sgRNA for expressing SgRNA and Cas9D10A at the same time is constructed; the plasmid PYSY-sgRNA is connected and transformed to an escherichia coli DH5alpha competent cell; finally, a transformant is verified; restriction enzyme digestion and sequencing identification prove that the construction of the CDK2 gene knockout vector is accurate. The vector constructed by CRISPR / Cas9 is adopted, and a theoretical basis is provided for subsequently obtaining a goat CDK2 gene deletion type cell line and researching a molecular mechanism of cell apoptosis molecules triggered by mycoplasma pneumonia infection.

The invention discloses a beta-carboline ruthenium compound. The molecular formula of the beta-carboline ruthenium compound is [Ru (N^N) 2 (Nh) XClY] (A^A)z, wherein N^N is selected from bpy or phen; A^A is selected from PF6 or SO3CF3; X is equal to 1 or 2, Y is equal to 2-X, and Z is equal to 1 or 2; or the molecular formula of the beta-carboline ruthenium compound is [Ru (N^N) 2 (1-Py-betaC)] (PF^)2, wherein N^N is selected from bpy, phen or DIP. On a molecular mechanism, the invention proves that the beta-carboline ruthenium compound can be used for treating cancers and has a better effectthan the effects of that of the traditional metal compound cisplatin with wide application and ruthenium compounds NAMI-A in clinical tests; . Thethe beta-carboline ruthenium compound for treating the cancers as an autophagic cell inducer fills the blank of the prior art and develops a new line for developing a new generation of high-efficiency medicaments for treating malignant tumors.

The invention relates to the molecular biology field and particularly relates to a gene knockout carrier, a construction method and application thereof and a gene knockout method of an NLRP1 gene of an MH7A cell. A CRISPR-Cas9 gene knockout system is utilized for carrying out CRISPR targeting sequence design by taking NLRP1 as a target gene so as to prepare a knockout carrier aiming at the NLRP1 gene, and the knockout carrier is utilized for transfecting the MH7A cell, so that the NLRP1 gene in the MH7A cell can be efficiently knocked out; and therefore, a research platform for rheumatoid arthritis is effectively built, so that the research of the pathogenesis of the rheumatoid arthritis is greatly promoted, and the researches of the interaction of NLRP1 inflammasomes and various inflammatory factors and the disease related molecular mechanisms of the rheumatoid arthritis and meningitis can be promoted.

This invention is in the field of homeostasis analysis. More particularly, it relates to systems and methods for analyzing persistent homeostatic perturbations, i.e. chronic stress, by measuring levels of biomarkers that are related to chronic stress. This invention is also directed to systems and methods for analyzing the molecular mechanisms of chronic stress.

The invention discloses a tumor cloning mutation detection method and device based on next-generation sequencing and a memory medium. The method provided by the invention comprises the steps of carrying out mutation detection on a comparison file of paired tumor and normal samples through utilization of mutation detection software, computing a mutation frequency, and selecting segments with high sequencing quality as a statistics result; carrying out copy number and purity detection on the paired tumor and normal samples through utilization of purity detection software; combining small segments into big segments, and annotating the copy number in a mutation area; and computing a proportion of the mutation in a tested tumor tissue through utilization of a beta distribution model according to tumor sample purity and copy number detection results, thereby judging a tumor cloning mutation type. According to the method provide by the invention, influences of the sample purity and multiploidon the detection are avoided, the mutation type detection is relatively accurate, the subcloning mutation with clinical significance can be effectively identified, and the foundation for accurately and deeply researching a tumor cloning evolution process and searching a tumor therapy molecular mechanism is laid.

Parkinson's disease is caused by the preferential loss of substantia nigra dopamine neurons. A Parkin Interacting Substrate, PARIS (ZNF746) is identified. The levels of PARIS are regulated by the ubiquitin proteasome system via binding to and ubiquitination by the E3 ubiquitin ligase, parkin. PARIS is a KRAB and zinc finger protein that accumulates in models of parkin inactivation and in human brain Parkinson's disease patients. PARIS represses the expression of the transcriptional co-activator, PGC-1α and the PGC-1α target gene, NRF-1 by binding to insulin response sequences in the PGC-1α promoter. Conditional knockout of parkin in adult animals leads to progressive loss of dopamine (DA) neurons that is PARIS dependent. Overexpression of PARIS causes selective loss of DA neurons in the substantia nigra, which is reversed by either parkin or PGC-1α co-expression. The identification of PARIS provides a molecular mechanism for neurodegeneration due to parkin inactivation.

The invention provide the method to enable the molecular detections of cellular fluid of at least a single cell easily and rapidly and clarify the dynamics of life phenomena and molecular mechanisms very rapidly by conducting simultaneous observations of cellular morphological dynamism. At the same time, it provides the method of exploring active molecules. Even in case that the target is a single cell, the cellular fluid is directly sucked by the capillary tip and the fluid is subjected to the analysis directly. By performing this method under the observation of the cell with image-magnifying scopes, such as a microscope, the cellular fluid is sucked and captured by the nanospray ionization capillary tip even if the target is a single cell and the trapped sample is subjected to the mass spectrometer by nano-spraying the sample solution directly after addition of the ionization supporting solvent into the tip, and then the molecular components in the cellular fluid is detected. The detected molecular peaks specific to a certain state are identified by performing differential analysis or t-test between the data groups of cells in different states.

The invention discloses a tomato S1lPIF4 gene, protein and application thereof to improving the low temperature resistance of plant. The nucleotide sequence of the S1lPIF4 gene is SEQ ID NO.1, and thecorresponding amino acid sequence is as shown in SEQ ID NO. 2. A tomato SlIPIF4 overexpression plant or a gene knockout plant through the gene means to regulate the expression level of the S1lPIF4 gene in order to do research on the regulatory mechanism of the S1lPIF4 gene on the low temperature resistance of tomato. According to the result, overexpression of S1lPIF4 at the lower temperature canpromote accumulation of plant hormone abscisic acid (ABA) and jasmonic acid (JA), inhibits accumulation of gibberellin (GA), further induces expression of the low temperature resistant gene of the tomato, and finally improves the low temperature resistance of the tomato. Therefore, the S1lPIF4 gene of the tomato enhances the low temperature resistance by inducing formation of hormones ABA and JA in the body. The tomato S1lPIF4 gene provides the gene resource for breeding new varieties of low temperature resistant tomato, has the good potential application value, and lays theoretical basis fordoing research on tomato plant distress response signal mechanism and molecular mechanism of being tolerant to adverse environment.

The invention relates to a method for inducing nematode-trapping fungi to synchronously produce trapping organs, which belongs to the field of applied microbiology. The method comprises the steps of culture of nematodes, preparation of a nematode extract, culture of nematode-trapping fungi and induction of trapping organs. The method can utilize the nematode extract to induce the nematode-trapping fungi to synchronously produce a large number of trapping organs (160 trapping organs / mg mycelium) by culturing the nematodes (Caenorhabditis elegans) and preparing the nematode extract by utilizing ultrasonic waves for crushing and centrifugalizing. The method has the advantages of simple and easy operation and good repeatability, and can obtain a large number of synchronous three-dimensional fungal nets, thereby providing good experimental materials for further research of the molecular mechanism of forming the trapping organs of the nematode-trapping fungi.

The invention relates to a wheat CBL-CIPK (CBL-interacting protein kinase) stress tolerance regulatory factor as well as an encoding gene and an application thereof. According to the wheat CBL-CIPK stress tolerance regulatory factor, protein kinase genes which are consciously induced by salt and drought when being expressed in roots and leaves are selected according to expression profile chip information of salt and drought stress of wheat, and a cDNA (complementary deoxyribonucleic acid) sequence with a whole-length encoding area is cloned, wherein the nucleotide sequence is as shown in SEQ ID NO.1. Analysis shows that the gene can encode CBL-CIPK serine / threonine protein kinase, the amino acid sequence is as shown in SEQ ID NO.2, and is named as TaCIPK33. Transgenic function analysis shows that the gene has the functions of improving plant salt tolerance and drought resistance. Basis is provided for further understanding molecular mechanisms of wheat in responding to drought and saltstress, and meanwhile the gene can be applied to genetic improvement of novel stress tolerance germplasms of crops such as wheat.

The invention belongs to the technical field of plant genetic engineering and in particular relates to pyrophosphatase of ammopiptanthus mongolicus, and a coding gene and application thereof. The pyrophosphatase provided by the invention comes from the ammopiptanthus mongolicus, named AmVP1, wherein the amino acid sequence of the pyrophosphatase is shown as SEQ ID NO.2. The coding gene of the pyrophosphatase of the ammopiptanthus mongolicus is one of the following nucleotide sequences: SEQ ID NO.1 and polynucleotide with the amino acid sequence shown as SEQ ID NO.2. The gene of the invention can be applied to the research of the salt-resisting and drought-enduring molecular mechanism of the ammopiptanthus mongolicus, and is used for improving the salt-resisting and drought-enduring property of plants.

The present invention is used for evaluating the advantages of haliotis discus hannai. A multi-phase HPLC purification system is used to purify anti-inflammatory peptide from abalone (AAIP, abalone anti-inflammatory peptide). In tandem MS analysis, a fragmentation result shows that the amino acid sequence of the AAIP with nitrogen monoxide (NO) inhibitory activity (IC50=55.8[um]M) is Pro-Phe-Asn-Glu-Gly-Thr-Phe-Ala-Ser (1175.2Da). While the anti-inflammatory effect of RAW264.7 macrophages generated by the stimulus of the AAIP to lipopolysaccharides (LPS) is further studied, and the molecular mechanism is elaborated. The result shows that the AAIP peptide inhibits the nitrogen monoxide (NO) generation induced by the LPS through the expression of inducible nitric oxide synthase (iNOS) by a dose-dependent manner, and the gene transcription of pro-inflammatory cytokines is also obviously reduced, wherein the pro-inflammatory cytokines comprise such as interleukin (IL-1 beta), tumor necrosis factors (TNF-beta) and IL-6. In addition, the AAIP obviously inhibits phosphorylation of mitogen-activated protein kinases (MAPK), such as p-p38 and p-JNK. These results indicate that the AAIP inhibits inflammatory response induced by LPS by intercepting the MAPK pathway of the macrophages. Therefore, the AAIP can be applied to therapeutic drugs for inflammations treatment or healthcare food products.

This invention provides a genetically tractable in situ non-human animal model for hepatocellular carcinoma. The model is useful, inter alia, in understanding the molecular mechanisms of liver cancer, in understanding the genetic alterations (e.g., in oncogenes and tumor suppressor genes) that lead to chemoresistance or poor prognosis, and in identifying and evaluating new therapies against hepatocellular carcinomas. The liver cancer model of this invention is made by altering hepatocytes to increase oncogene expression, to reduce tumor suppressor gene expression or both, preferably by inducible, reversible, and / or tissue specific expression of double-stranded RNA molecules that interfere with the expression of a target gene, and by transplanting the resulting hepatocytes into a recipient non-human animal. The invention further provides a method to treat cancer involving cooperative interactions between a tumor cell senescence program and the innate immune system.

The invention discloses effector protein Avh87 derived from phytophthora cinnamomi, and a coding gene and application thereof. The protein has the amino acid sequence of SEQ ID No. 1 in the sequence table. The nucleotide sequence expressing the protein is represented by SEQ ID No. 2 in the sequence table. Experiments show that the gene can inhibit plant cell hypersensitive reactions induced by apoptotic precursor protein Bax, or inhibit plant cell death induced by pathogenic phytophthora elicitors INF1. The effector protein Avh87 is of great significance in enriching molecular mechanism information of the interaction between the phytopathogenic oomycetes and hosts, and in establishing a comprehensive prevention and control technology strategy for phytopathogenic oomycete diseases.

Adult neural stem cells differentiate into neurons, astrocytes, and oligodendrocytes in the mammalian CNS, but the molecular mechanisms that control their differentiation are not yet well understood. Insulin-like growth factor-I (IGF-I) can promote the differentiation of cells already committed to an oligodendroglial lineage during development. However, it is unclear whether IGF-I affects multipotent neural stem cells. Here we show that IGF-I stimulates the differentiation of multipotent adult rat hippocampus-derived neural progenitor cells into oligodendrocytes. Modeling analysis indicates that the actions of IGF-I are instructive. Oligodendrocyte differentiation by IGF-I appears to be mediated through an inhibition of BMP signaling. Furthermore, overexpression of IGF-I in the hippocampus leads to an increase in oligodendrocyte markers. These data demonstrate the existence of a single molecule, IGF-I, that can influence the fate choice of multipotent adult neural progenitor cells to an oligodendroglial lineage.

The invention discloses a miRNA expression profile model related to human multiple myeloma, a construction method and an application thereof. The invention discloses a miRNAs expression profile of tumor cells and circulating body fluid of a patient suffering from multiple myeloma, determines a molecular map of differentially expressed miRNAs between the patient suffering from multiple myeloma andhealthy people, and screens a biological marker for early diagnosis, prognosis evaluation and curative effect prediction of myeloma diseases, analyzes the myeloma cell characteristic miRNAs molecularexpression profile, correspondingly regulates mRNA in a targeted manner, and explores the correlation between differential miRNA and mRNA molecules and myeloma disease progression and the molecular mechanism of the correlation. The miRNAs expression profile disclosed by the invention provides a theoretical basis and an experimental basis for the treatment of multiple myeloma.

The invention discloses a method and a system for accurately analyzing a DMD gene structural variation breakpoint. The method includes steps of receiving of fragment sequences, initial comparison, repeat comparison, position analysis, analysis of suspected breakpoint, and in-depth correction; through the method, the type and the breakpoint position of DMD gene copy number variation can be accurately analyzed; the mean detection bias rate is less than 4 bp; the accuracy is high, and the stability is good; besides, the method and the system can cover the variation detection of an exon and an introne at the same time, solve the shortcoming that the breakpoint of the gene copy number variation cannot be accurately analyzed in the prior art, and provide technical support for realizing the rapid and parallel DMD breakpoint detection service and the study of the DMD structure gene variation molecular mechanism.

The invention discloses a VIGS silence system for identifying a peony UDPglucose Flavonoid Glucosyl-Transferase gene. The VIGS silence system for identifying a peony UDPglucose Flavonoid Glucosyl-Transferase gene comprises a recombinant vector which is obtained by adding desired genes to a VIGS silence vector. The desired genes are nucleotide sequences shown as a first sequence in a sequence table and / and nucleotide sequences shown as a third sequence in the sequence table. The UDPglucose Flavonoid Glucosyl-Transferase gene PsUF3GT and a PsUF5GT conserved segment are cloned by homology-based cloning technologies as target genes to construct TRV2 vector, so that the VIGS silence system is built successfully, and amount of gene expression of the UDPglucose Flavonoid Glucosyl-Transferase gene PsUF3GT and the PsUF5GT conserved segment and total anthocyanin content are reduced. The system has great significance for peony genes function study and lays foundation for analyzing an molecular mechanism formed by peony splash.

The present invention is the method of establishing mouse model of viral hepatitis. The method includes selection of mouse hepatitis virus-III (MHV-3) of 100-1000 pFu, injecting infection of variousvariety mice of BALB / CJ, C3H / HEJ and A / J mice, and observing the survival rate and liver tissue change. Via the said process, three kinds of viral hepatitis mouse model are established, of fulminanthepatitis, chronic hepatitis and symptom-free mouse separately. The establishment of the model provides powerful facilities for systematic research of viral hepatitis in genetic regulation, molecular mechanism and prevention and treatment.

A method for differentiating or promoting or inducing differentiation of stem cells into pluripotent cardiovascular progenitors (MCPs) by transiently inducing the expression of a single gene, namely Mesp1, is disclosed. Cells obtained by the method and their uses in research and clinical settings are also disclosed. Using genome wide transcriptional analysis, the inventors found that Mesp1 rapidly activates and represses a discrete set of genes, which form potential new targets for both therapy and for the identification of MCPs. Insights into the molecular mechanisms underlying the earliest step of cardiovascular specification and potential methods for dramatically increasing the number of cardiovascular cells for cellular therapy in humans are provided.

The invention discloses a section of gene and its protein with controlling rice tillering angulations, and its application. The purpose is that providing a section of gene and its protein with controlling rice tillering angulations, and applying the gene and protein in controlling rice tillering angulations. The gene's cDNA has one of nucleotide sequences as follow: 1) the SEQ ID NO: 2 or SEQ ID NO: 4 DNA sequence in sequence table; 2) coding the DNA sequence of SEQ ID NO: 1 in the sequence table; 3) the nucleotide sequence that can hybrid with restrict DNA sequence of SEQ ID NO: 2 in the sequence table at high strict condition. The gene and its protein have important theory and operation significance in studying the molecular mechanism of rice tillering angulations and controlling rice plant type. The invention has extensive application and market outlook in agriculture field.

The invention relates to the technical field of gene knockout and particularly discloses a gene knockout method for breeding selection of a fhl1b gene-deletion type zebra fish. According to the method, by means of a CRISPR / Cas9 gene editing technique, an appropriate target locus is designed on a fhl1b gene of the zebra fish, specific sgRNA and Cas9-mRNA which are synthesized outside the fish bodyare microscopically injected into one cell of the zebra fish, and 60 hours after embryo cultivation is carried out, by selecting embryos to conduct genotype analysis, the effectiveness of the selectedlocus is proved. The off-target rate is low, not only is the fhl1b gene removed by interference, but also more convenience is provided for further disclosing the whole process of generation of a heart shape and regulating and controlling a molecular mechanism of the process by adopting a genetic approach to studying the functions of the fhl1b gene, and the method has a significant meaning in understanding of medical pathology of heart diseases and research and development on new therapeutic schemes.