130 results about "Fibre cell" patented technology

The invention provides a production method for Fbxo40 gene knockout pigs. The production method comprises the steps that a CRISPR-Cas9 targeting vector and a PGK-Neo resistant gene are jointly transfected into a porcine embryonic fibroblast to obtain a G418-resistant positive monoclonal cell, an Fbxo40 gene in the positive monoclonal cell generates insertion or deletion mutation, a reading frame generates frame shift and stops in advance, then the obtained cell is cloned to serve as a donor cell for nuclear transfer, and an oocyte serves as a receptor oocyte for nuclear transfer; cloned embryos are obtained through a somatic cell nuclear transfer technique, the high-quality cloned embryos are transferred into the fallopian tube of an oestrous sow, and the Fbxo40 gene knockout pigs are obtained through whole-period development. According to the production method, the Fbxo40 knockout pigs are efficiently obtained through a CRISPR-Cas9 gene editing technique at the low cost, and animal models are supplied to research of muscle development and diseases related to the muscles.

The present invention relates to the identification of new proteins comprising fibroblast growth factor 21 (FGF21) and other metabolic regulators, including variants thereof, known to improve metabolic profiles in subjects to whom they are administered. Also disclosed are methods for treating FGF21-associated disorders, GLP-1-associated disorders, and Exendin-4-associated disorders, including metabolic conditions.

The present invention relates to mutants of Fibroblast Growth Factor (FGF), particularly FGF-20 and FGF-21, which contain newly introduced N-linked or O-linked glycosylation site(s). The polynucleotide coding sequences for the mutants, expression cassettes comprising the coding sequences, cells expressing the mutants, and methods for producing the mutants are also disclosed. Further disclosed are pharmaceutical compositions comprising the mutants and method for using the mutants.

The invention provides compositions and methods that employ compounds that can stimulate proliferation of fibroblasts or keratinocytes and / or stimulate production of collagen by fibroblasts. These compositions and methods are useful for treating gum- and skin-related conditions.

The invention discloses a method for improving the ash content of paper-making process reconstituted tobaccos. The method includes the steps that after tobacco raw materials are extracted, solid-liquid separation is carried out, tobacco liquid is partially refined, concentrated and charged and then is manufactured into coating liquid, pulping treatment is partially conducted on the tobacco solid part, so that tobacco slurry fibers are obtained, CaCO3 is added into the tobacco slurry fibers, and the CaCO3 is filled into cell cavities of the tobacco slurry fibers through pits of the tobacco slurry fibers; afterwards, the tobacco slurry fibers and additional fiber slurry are mixed according to the weight ratio of 4 to 1, and then paper manufacturing with pulp is carried out to manufacture reconstituted tobacco sheets; the coating liquid is made to coat the reconstituted tobacco sheets, so that the paper-making process reconstituted tobaccos are manufactured. According to the method, the technology that the CaCO3 is filled into the cell cavities of the tobacco slurry fibers is applied to the preparation technology of the paper-making process reconstituted tobaccos, any chemical agent such as a high-molecular polymer is not added, and the purposes that the ash content is improved and meanwhile the strength performance of the paper-making process reconstituted tobacco sheets is improved can be achieved.

The invention provides compositions and methods that employ compounds that can promote skin cell renewal, wound healing, proliferation of fibroblasts and / or keratinocytes, and production of collagen or fibronectin by fibroblasts. These compositions and methods are useful for rejuvenating the skin and for treating wounds as well as gum-related and skin-related conditions.

The present invention relates to the identification of new proteins comprising fibroblast growth factor 21 (FGF21) and other metabolic regulators, including variants thereof, known to improve metabolic profiles in subjects to whom they are administered. Also disclosed are methods for treating FGF21-associated disorders, GLP-1-associated disorders, and Exendin-4-associated disorders, including metabolic conditions.

The c-Myc oncoprotein, a helix-loop-helix-leucine zipper (HLH-ZIP) transcription factor, is frequently deregulated in human cancers. All known functions of c-Myc, including those pertaining to transformation, require that it heterodimerize with another HLH-ZIP protein, Max. Using a high throughput yeast-based assay, we identified seven low molecular weight substances that inhibit c-Myc-Max association. Each compound also prevented this interaction in vitro and inhibited the growth of c-Myc-expressing fibroblasts, although not of fibroblasts lacking c-Myc. Finally, short-term exposure of c-Myc over expressing fibroblasts to several of the compounds markedly reduced their in vivo tumorigenicity. These studies suggest that yeast-based assays can be used to identify inhibitors of protein-protein interactions and that these frequently function in mammalian cells. The signature specificities of each of the c-Myc-Max compounds identified here further suggest synergistic in vivo function.

Described herein are devices and methods for extracting cellular material from living cells and then depositing them into to a receptacle in a nanoliter scale. Using a nanopipette integrated into a scanning ion conductance microscope (SICM), extraction of mitochondrial DNA from human BJ fibroblasts and Green Fluorescent Protein (GFP) transcripts from HeLa / GFP cells was achieved with minimal disruption to the cellular milieu and without chemical treatment prior to obtaining the isolated sample. Success of the extraction was confirmed by fluorescence microscopy and PCR analysis of the extracted material. The method and apparatus may be applied to many different cell types and intracellular targets, allowing not only single cell analysis, but single subcellular compartment analysis of materials extracted in their native state.

The invention provides novel pharmaceutical purposes of jaceosidin and specifically relates to application of jaceosidin in prevention or treatment of pulmonary fibrosis, wherein through inhibition of pulmonary epithelial-mesenchymal transition, pulmonary vascular endothelial-mesenchymal transition and propagation of fibroblasts, growth and development of pulmonary fibrosis are inhibited, so prevention or treatment of pulmonary fibrosis is realized. In the above-mentioned application, the dosage range of applied jaceosidin is 20 to 1000 mg, preferably, 20 to 500 mg.

The invention relates to a cotton GhFP1 gene which belongs to a member of a bHLH transcription factor gene family, the cDNA (complementary deoxyribonucleic acid) full length is 1262bp, and the cotton GhFP1 gene can code a bHLH protein containing 355 amino acids. GhFP1 is respectively expressed in various tissues of cotton, and is highly expressed in the fiber development premetaphase. The GhFP1 promoter segment with the length of 1061bp is separated and cloned. The analysis on the activity of the promoter by using the GUS report gene indicates that the promoter has higher specific expression activity in cotton fibers. The experiment indicates that the overexpression of GhFP1 in Arabidopsis thaliana results in increase of leaf epidermal hair; and the overexpression of GhFP1 in cotton results in enhancement of expression of related genes of cotton fibrocyte brassinolide and increase of ripe cotton fiber length. The result proves that the GhFP1 protein performs positive regulating actions and promotes elongation of fibrocytes in the cotton fiber elongation process, and thus, possibly has important application value in improving the cotton fiber quality and increasing the cotton yield.

The invention discloses a preparation method for a myostatin knock-out pig, belonging to the technical field of biology. The preparation method is as follows: generating virus particles capable of expressing shRNA of myostatin gene of a targeted pig in a 293Ft cell; then infecting the fibroblasts of a piglet by using the produced slow virus; wherein after 48 hours, the expression of Myostatin genes in the fibroblasts is inhibited by fluorescence quantitative polymerase chain reaction (PCR) detection. In the invention, the slow virus particles can be used for carrying out microinjection on perivitelline space of a fertilized egg of the pig, can lead the virus nucleic acid to be integrated to an embryo gene group so as to express the shRNA, and can produce a transgenic pig with the myostatin of being knocked out after embryo transplantation. The slow virus particles also can be used for producing the transgenic pig by using a sperm vector method. By utilizing the preparation method, the cumbersome operation of knocking out the homologous targeting gene in the traditional method can be avoided, the virus consumption is less and the stable passage can be achieved.

The invention relates to preparation and application of an efficient cell inactivation vaccine against new duck reovirus disease. The preparation method comprises the following steps: an antigen preparation step: inoculating a virus NDRV-TH11 strain with immortalized duck embryo fibroblast to obtain cytotoxicity with 80% of cytopathic effect, and performing cryopreservation at -20 DEG C; and a vaccine preparation step: inactivating the obtained cytotoxicity and mixing with a homemade nano adjuvant and stirring uniformly to obtain the efficient cell inactivation vaccine against new duck reovirus disease. After the duck is immunized by the efficient cell inactivation vaccine against new duck reovirus disease prepared in the invention, the duck can effectively resist the attack of a virulent strain of the new duck reovirus.

The invention describes methods and agents for inducing in the skin one or more effects selected from stimulating fibroblast migration, stimulating elastin production, reducing expression of inflammatory factors and upregulating specific genes, thereby preventing skin cell aging processes and repairing skin damage. In preferred embodiments, the methods and agents comprise active extracts produced from fish eggs.

Specific binding members, particularly antibodies and fragments thereof, which bind to Fibroblast Activation Protein (FAP) are provided, particularly recognizing both human and mouse FAP. These antibodies are useful in the diagnosis and treatment of conditions associated with activated stroma, including wound healing, epithelial cancers, osteoarthritis, rheumatoid arthritis, cirrhosis and pulmonary fibrosis. The anti-FAP antibodies, variable regions or CDR domain sequences thereof, and fragments thereof may also be used in therapy in combination with chemotherapeutics, immune modulators, or anti-cancer agents and / or with other antibodies or fragments thereof. Antibodies of this type are exemplified by the novel antibodies ESC1 1 and ESC 14 whose sequences are provided herein.

The invention discloses RNA and application of RNA in diseases of a cardiovascular system. According to RNA, the nucleotide sequence of RNA is a sequence 3 in a sequence table. The invention further provides a gene,encoding RNA. The nucleotide sequence of the gene of RNA is a sequence 4 in the sequence table. The experiment of the invention shows that RNA and the application of RNA in the diseases of the cardiovascular system find that by an induction of isoproterenol (ISO), RNAi method Knock down IHP-55 is used in neonatal rat cardiac fibroblasts to obviously promote proliferation of cardiac fibroblasts; ISO induced activation and kinase activity of P38MAPK can be enhanced in the Knock-down HIP-55 cardiac fibroblasts; and HIP-55 is a new protein which has a protective effect on cardiac fibrosis, and provides a new target for clinical diagnosis, treatment and new medicament development.

The invention provides a method for knocking out a CRISPR / Cas9 mediated sheep FGF5 gene and integrating an MTNR1A gene at a fixed point.The method includes the steps of transferring the CRISPR / Cas9 targeting vector of the specific targeting third exon of the sheep FGF5 gene and the gene homologous recombination vector into a sheep fetus fibroblast to obtain a sheep FGF5 gene knockout cell with a fixed-point integration of the MTNR1A gene. The donor vector and the targeting vector are co-transfected into sheep fetus fibroblasts in a nuclear transfer mode, and specific expression of MTNR1A in sheep ovary tissue can be achieved through a CYP17 promoter in the unmarked donor vector constructed through a Gibson Assembly method. The separated and identified positive monoclonal cell strain can beused as a nuclear donor for nuclear transplantation of somatic cells to obtain cloned embryos, so that a solid foundation is laid for culturing transgenic clone sheep which locally strengthens melatonin signals and produces more hairs in a reproductive system.

The invention belongs to the technical field of duck plague virus culturing, particularly relates to a new host cell of the duck plague virus, and discloses application of a chicken liver cancer cell system as a duck plague virus host. A traditional method for culturing the duck plague virus through the duck embryo fibroblast primary cell is limited by duck embryo supply and the hatching day age, and wastes time and labor during preparation. The established method for culturing the duck plague virus chicken liver cancer cell system is good in virus growth. The method for culturing the virus through a continuous cell line is not influenced by duck embryo supply, and has the advantages of being instantly available and capable of saving time and labor. The duck plague virus can grow and reproduce on a chicken liver cancer cell system continuous cell culture medium, and by means of a newly-found virus copying host cell, a new technical approach, method and material are provided for research of related fields.

The invention provides lung adenocarcinoma exosome specific miRNAs, and a target gene and applications thereof. High flux sequencing technology is combined with systematic bioinformatics analysis methods, the specific high-content exosome miRNAs related with lung adenocarcinoma is obtained through screening; the RNA sequences of the lung adenocarcinoma exosome specific miRNAs are represented by SEQ ID No.1-3. It is found that the common target gene of the miRNAs is FOXN3. The target gene is capable of inhibiting fibroblast migration capability, reduction of the expression amount of the targetgene is capable of promoting entering of fibroblast into tumor tissues, and promoting tumor growth. The lung adenocarcinoma exosome specific miRNAs and the target gene thereof can be taken as lung adenocarcinoma diagnosis markers and can be used for preparation of diagnostic kits. An inhibitor of the miRNAs or an expression promoter of the target gene can be used for preparing medicines used for treating lung adenocarcinoma, or medication guidance in treatment of lung adenocarcinoma; and the clinical application value is relatively high, and the application prospect is promising.

The invention discloses eye essence as well as a preparation method and an application thereof, relates to the technical field of skin care products and aims to improve a nursing effect on eye skin and effectively solve the problems of saggy eye skin and proneness to wrinkles. The eye essence contains palmitoyl tripeptide-5, algae extract* / pullulan / water, sodium hyaluronate, panthenol, epidermal growth factor and other substances and can provide nutrients for skin, stimulate fibroblast proliferation and collagen synthesis, promote skin regeneration and tighten the skin, wherein palmitoyl tripeptide-5 can be beneficial to rapid relaxing of mimetic muscle, and a wrinkle relieving effect is realized; algae extract* / pullulan / water can ensure that a physical film is formed on the skin surface instantly by the eye essence, the periocular skin is tightened instantly, and the instantaneous wrinkle relieving effect is enhanced. The eye essence can effectively improve the nursing effect on the eye skin and delay eye skin aging, and beautiful appearance is maintained for a longer time.

The invention belongs to the technical field of pig transgenosis and particularly relates to a method for producing transgenic pigs through overexpression HOXA10 genes. The method is characterized in that constructed HOXA10 gene expression carriers are subjected to lipofection and sulfate (G418) screening to obtain HOXA10 transgenic cell lines with stable expression; the HOXA10 transgenic cell lines are transplanted into denucleated pig oocytes with HOXA10 transgenic pig fibroblasts as nuclear donors through an artificial nuclear transplantation method to form reconstructed embryos; the reconstructed embryos are transplanted into a receptor sow oviduct, and after a sow gives birth to the HOXA10 transgenic pigs and molecular identification is conducted, masculine HOXA10 transgenic pigs are screened out. The transgenic pigs obtained through the method provide materials for research about the mechanism of improving the reproductive capacity through the HOXA10 genes and provide an animal model for revealing other functions of the HOXA10 genes.

The invention belongs to the field of daily chemicals, and particularly relates to a skin-care composition with a sunscreen effect as well as a preparation method and application thereof. The skin-care composition, provided by the invention, comprises the following components: a natural plant extract, palmitoyl tetrapeptide-7, vitamin E, oryzanol and oligomeric proanthocyanidin, wherein the natural plant extract contains a variety of natural plant sunscreen extracts, has a very good overall effect of absorbing ultraviolet light, scavenging free radicals, inhibiting generation of melanin, inhibiting release of allergy mediators, protecting fibroblasts, inhibiting the activity of matrix metalloproteinase and the like, and has a very good sunscreen effect; the oligomeric proanthocyanidin can be used as a protective agent for the oryzanol and the vitamin E, is free of skin irritation, has an antioxidant protective effect on skin, and can improve the skin-care and sunscreen effect of the composition.

The invention discloses a method for preparing foot-and-mouth disease virus-resistant (FMDV) transgenic livestock by expressing FMDV-resistant ribonucleic acid interference (RNAi), which comprises the following steps of: linearizing an FMDV-resistant RNAi recombinant Lenti-virus vector which carries a short hairpin RNA (shRNA) base sequence and a green fluorescent protein (GFP) reporter gene and transfecting fetus fibroblasts of normal livestock; sieving transgenic nuclear donor cells by using the GFP reporter gene and a flow cell sorter and introducing nuclear donor nuclei into denucleated oocytes of the livestock to obtain a reconstructed embryo; and immigrating the reconstructed embryo into the uterus of surrogate livestock to obtain an offspring, namely the FMDV-resistant RNAi heterozygote transgenic livestock. Heterozygote transgenic male livestock and heterozygote transgenic female livestock are normally mated or reproduced by artificial insemination so as to obtain homozygote transgenic livestock. The transgenic livestock prepared by the method can express the FMDV-resistant RNAi, obviously reduces the FMDV infection rate and has the capability of resisting foot-and-mouth disease (FMD).

A method for reducing or preventing the formation of scar tissue is disclosed. The method comprises applying a pharmacologically effective amount of a composition comprising about 5.0% by weight of ovalbumin, about 1.0% phenoxyethanol, about 0.5% carbomer, and about 0.3% triethanolamine. A method for stimulating fibroblast production is also disclosed. The method comprises applying a pharmacologically effective amount of a composition comprising about 5.0% by weight of ovalbumin, about 1.0% phenoxyethanol, about 0.5% carbomer, and about 0.3% triethanolamine.

Fibroblast growth factor-18 (FGF-18) binds with FGF receptors-2 and -3. Compositions comprising an FGF-18 component can be used to target cells that express these receptors. Suitable targets include tumor cells that express constitutively activated forms of FGF receptors-2 and -3. For example, conjugates of FGF-18 and saporin can be used to target tumor cells that express FGF receptors-2 or -3, and to inhibit the proliferation of these cells.

The invention discloses a method for breeding a transgenic pig for the over expression of a pig PBD-2 gene. The method comprises the following steps: (1) building and straightening a pc DNA 3.1 (+) / pCA-PBD2 carrier in such a manner that a primer is designed, the liver tissue RNA of a pig is extracted, RT-PCR is enlarged to obtain a PBD-2 gene, alcohol is used for precipitation, and sterilization water is used for dissolution; (2) transfecting lipidosome and screening cells in such a manner that a pig fibroblast for over-expression of the PBD-2 is built; (3), obtaining a recombined blastula according to a manual clone method; (4), transplanting an embryo in such a manner that the bred blastula is transplanted into the fallopian tube of a surrogate sow; and (5), identifying the transgenic pig. The method has the advantage that the operation is feasible, simple and convenient. After being cloned, the pig PBD-2 gene is inserted into the expression vector, and a pig fibroblast over expressing the pig PBD-2 is screened out through cell transfection; the pig fibroblast is transplanted to the megagametocyte of the sow through manual cloning to obtain a reconstructed blastula; and a masculine blastula is transplanted to the fallopian tube of the recipient sow to obtain the transgenic pig for over-expression of the PBD-2 gene. The method has a wide anti-bacterial capability, and provides a favorable material for disease-resistant breeding research.

The invention relates to a production method for recombinant human fibroblast growth factor-18 protein. The production method comprises the following steps: 1, expressing the fibroblast growth factor-18 protein in insect cells by using an insect baculovirus vector expression system, specifically comprising 1) obtaining recombinant baculovirus Bacmid-FGF18 (fibroblast growth factor-18), wherein the FGF18 gene is a fibroblast growth factor-18 gene and has a base sequence as shown in SEQ ID NO: 1, and 2) infecting the insect cells with the Bacmid-FGF18 recombinant baculovirus with MOI (Multiplicity of Infection) of 1 to 32, and then culturing the infected insect cells for 12 to 96 hours at the temperature of 25 to 29 DEG C to harvest cell fluid; and 2, purifying the cell fluid harvested in the step 1 to obtain the human fibroblast growth factor-18 protein. The invention also discloses application of the human fibroblast growth factor-18 protein in treating bone diseases and alopecia.

The invention relates to a method for preparing an infectious bursal disease virus live vaccine by utilizing a passage chicken embryo fibroblast. By utilizing the method, the IBDV (infectious bursal disease virus) live vaccine is prepared by steps of preparing a production virus seed, preparing a vaccine preparing material, preparing an IBDV cell virus solution, inspecting a finished product, preparing a vaccine, performing split charging, freeze-drying, and the like. According to the method, the existing production method is improved in aspects of inoculation material, culture solution improvement, culture environment maintenance, freeze-drying of protective agent components, and the like, and a new passage cell culture method is originally created. In addition, the invention also comprises an application of the IBDV live vaccine prepared by the method in prevention of an infectious bursal disease. The IBDV live vaccine prepared by the method has the characteristics of good heat resistance, high virus titer and good stability and is suitable for large-scale vaccine production.

Compositions of matter, of production, and treatment modalities are disclosed for the prevention and / or therapeutic reduction of tumors through induction of immunity against tumor associated fibroblasts and components of tumor microenvironment. In one embodiment of the invention, placentally derived fibroblast cells are cultured under conditions replicating tumor microenvironment. Expression of CD248 on said cultured fibroblasts is used as a marker of effective manipulation. Cells expressing CD248 are utilized a immunogens for stimulation of immunity towards cancer associated fibroblastic cells.