49results about How to "No lethal effect" patented technology

The invention provides a nucleic acid molecule for encoding a chimeric antigen receptor targeting CD22. The chimeric antigen receptor comprises an extracellular region, a transmembrane region and a intracellular signal transduction region, the extracellular region encoded by the nucleic acid molecule comprises a CD22 binding domain, and the CD22 binding domain is amino acid sequence as shown in SEQ ID No. 3. By using flow cytometry, a degranulation analysis experiment and ELISA to detect cell factors secreted by a T cell, a fact that the CAR-T cell has a high lethal effect on B cell lymphoma cells and acute B cell lymphoma cell leukemia cells expressing CD22 and hardly has a lethal effect on cells which do not express CD22, and an off-target effect is prevented effectively is proved. The chimeric antigen receptor CD22scFv-CD8alpha-4-1BB-CD3zeta can be used for treating CD22<+>B cell hematologic neoplasms and is applicable to combined treatment with a CD19 CAR-T cell.

The invention discloses a preparation method and application of a drug-loaded magnetic composite nanometer material. The fat-soluble solution (or emulsion) of the surface lipophilic magnetic nanoparticles and the drug is added to the dichloromethane solution of polylactic acid / glycolic acid copolymer (PLGA), and the three are mixed with each other to form an oil phase system. The oil phase system is added dropwise to the aqueous solution in which the surfactant Pluronic F127 is dissolved to form an oil-in-water system, and then the drug-loaded magnetic composite nanomaterial is obtained under the action of ultrasound. The drug-loaded magnetic composite nanomaterial can concentrate the drug in the tumor area under the action of an in vitro magnetic field to achieve targeting; the coating of PLGA solves the problem of mutual self-aggregation of magnetic particles, and the slow release control of the drug solves the problem It has great toxicity and side effects in the body and can be quickly taken up by tumor cells, leading to apoptosis of tumor cells and reversing the multidrug resistance of tumor cells.

The invention relates to an integrated ecological tomato cultivation method for reducing pest and disease damage and improving fruit quality. The method comprises the steps that nutrient solution diluent special for tomatoes is irrigated in a dripped mode 14-18 days after tomato seedlings are planted in a fixed mode, and diluted microorganism bacterium agents are sprayed during the fixed planting period of the tomato seedlings and / or the flowering-fruit expanding period. By means of the organic matrix, tomato special nutrient solution and microorganism bacterium agent integrated cultivation method, pest and disease damage to the tomatoes is reduced, the yield of the tomatoes is increased, and the quality of the tomatoes is improved. The tomato special nutrient solution has no lethal effect on the microorganism bacterium agents and provides nutrition for life activity of functional bacteria, by means of mutual collaboration, the yield of tomato fruit is increased, and the quality is improved.

The invention relates to applications of nicotinamide adenine dinucleotide in the field of pharmacy, and more specifically relates to applications of nicotinamide adenine dinucleotide in preparation of a medicine used for curing liver damages caused by chemotherapy drug doxorubicin hydrochloride. NAD<+> is capable of preventing and treating liver damages caused by chemotherapy drug doxorubicin hydrochloride effectively; and it is also found that NAD<+> is capable of killing glioma cells, neuronal tumor cells, gastric cancer cells, renal tumor cells or breast tumor cells effectively, but possesses no killing effect on normal primary cells.

The invention discloses target specific double mutant fusion protein, which is established by human gonadotropin releasing hormone mutant (mGnRH) and recombinant pseudomonas aeruginosa exotoxin A mutant (PE38m4a) fusion protein. The mGnRH-PE38m4a protein provided by the invention has obvious specific binding activity and cytotoxicity to the tumor cell lines which comprise colon cancer HT-29 cells, oophoroma OVCAR3 cells, cervical adenocarcinoma HeLa cells or liver cancer HepG-2 cells, and the like; the normal cells basically have no lethal function; the activity of the fusion protein is enhanced by approximate nine to ten times compared with the activity of natural PEA; the cost is low, the method is simple, therefore the target specific double mutant fusion protein is more suitable for industrialized production.

The present invention provides a nucleic acid molecule encoding a chimeric antigen receptor targeting CD5, the chimeric antigen receptor comprises an extracellular domain, a transmembrane domain and an intracellular signal transduction domain, and the extracellular domain encoded by the chimeric antigen receptor comprises a CD5 binding domain which is an amino acid sequence as shown in SEQ ID NO:3. The chimeric antigen receptor comprises an extracellular domain, a transmembrane domain and an intracellular signal transduction domain, and the extracellular domain encoded by the chimeric antigenreceptor comprises a CD5 binding domain which is an amino acid sequence shown in SEQ ID NO:3. The cytokines secreted by NK cells were detected by flow cytometry, degranulation assay and ELISA. The results showed that the CD5 CAR NK cells had strong cytotoxic effect on hematological tumor cells expressing CD5, but had weak cytotoxic effect on cells not expressing CD5. The cytokines secreted by CD5CAR NK cells could effectively prevent the missed target effect. A chimeric antigen receptor CD5 scFv-CD8[alpha]-4-1BB-CD 3 Xi can be used in the treatment of lymphocytic hematologic tumors with positive CD 5.

The invention relates to an integrated ecological zucchini cultivation method for reducing pest and disease damage and improving fruit quality. The method comprises the steps that nutrient solution diluent special for zucchinis is irrigated in a dripped mode 14-18 days after zucchini seedlings are planted in a fixed mode, and diluted microorganism bacterium agents are sprayed during the fixed planting period of the zucchini seedlings and / or the flowering-fruit expanding period. By means of the organic matrix, zucchini special nutrient solution and microorganism bacterium agent integrated cultivation method, pest and disease damage to the zucchinis is reduced. The zucchini special nutrient solution has no lethal effect on the microorganism bacterium agents and provides a nutrition condition for life activity of functional bacteria, by means of mutual collaboration, the quality of zucchini fruit cultivated through facilities is improved, and the yield is increased.

A process for removing nitrite and ammoniacal nitrogen in aquatic products cultivation includes loading H2O2 into an open container, enclosing the container by an osmotic film, attaching the container to an oxygen-enriching machine in water of pond, turning on the machine for slowly diffusing the H2O2 to decompose it into H2O and O2, oxidizing the nitrite and ammoniacal nitrogen to become nitrate by said O2, and adding photosynthesizing bacteria to absorb the nitrate.

The invention relates to the field of biological medicines, in particular to a single-chain antibody specifically bound with CLDN18.2 and a chimeric antigen receptor (CAR) containing the single-chain antibody. The invention also relates to reconstructed immune cells expressing the CAR, or co-expressing the CAR and a further bioactive molecule (e.g., a PD-1 antibody or mIL-15), nucleic acid molecules encoding such CAR or co-expressed molecules, and methods of preparing the reconstructed immune cells. The invention also relates to application of the CAR and the immune cells in preventing and / or treating cancers such as gastric cancer, gastric adenocarcinoma and pancreatic cancer and a method for preventing and / or treating the cancers such as the gastric cancer, the gastric adenocarcinoma and the pancreatic cancer.

The invention discloses a chorthippus microspore strain and applications thereof. The chorthippus microspore strain disclosed by the invention is paranosema acridophagus PA-201301 with the preservation number of CGMCC No.7757 in the China General Microbiological Culture Collection Center. The paranosema acridophagus PA-201301 CGMCC No.7757 disclosed by the invention has the advantages of obvious lethal effect to chorthippus dubius, chorthippus fallax, myrmeleotettix palpalis, calliptamus abbreviatus, bryodema luctuosum, oriental migratory locust and gastrimargus marmoratus than that of an existing locust microspore, and stress resistance, in particular strong ultraviolet resistance, wherein the germination rate of spores is still more than 80% after a spore solution is irradiated by the ultraviolet light for one hour; in addition, the production technology of microspores of the chorthippus microspore strain is simple, and propagation hosts are easily obtained; after homogenating, separation is performed at the centrifugal force of about 6000g; the chorthippus microspore strain disclosed by the invention is conductive to mass production and applications.

The invention relates to a preparation method of a composite magnetic nano photosensitizer. The preparation method includes preparing Fe3O4 magnetic nanoparticles, using tetraethoxysilane as a modifier to coat the surface of the Fe3O4 magnetic nanoparticles with a thin layer of silica, and fitting quantum dots CdT and a photosensitizer on the surface under the adsorbing action of silica mesoporousmicrospheres to obtain magnetic nano photosensitizer particles Fe3O4@SiO2@Qdots / PS. Specific bioluminescence of tumor cells can be used as a light source; the photosensitizer is excited to generate active oxygen clusters and convert electricity energy of an alternating electric field into thermal energy; the composite magnetic nano photosensitizer is a novel multifunctional photosensitizer.

The invetnio discloses a mastitis-proof cow breeding method, which comprises the following steps: constructing antibiotic peptide gene into astopic expressive carrier of galactophore tissue; extracting external gene to transmit and infect supplying cell; making fiber cell in the core-transplanting supplying cell; sieving to obtain positive cell and stoning oocyte to construct embryo; activating the restructed embryo to culture; transplanting normal breeding embryo into cow uterus synchronuously; obtaining gene-transmitting cloned cow.

The invention relates to the technical field of biology, in particular to a migratory locust Rab5 gene and application of dsRNA of the migratory locust Rab5 gene in migratory locust prevention and treatment. Specifically, through a bioinformatics method, a fragment of the migratory locust small GTP enzyme gene 5 is obtained from a migratory locust transcriptome, a primer corresponding to the fragment is further designed and synthesized, PCR amplification is carried out, and after sequencing of an amplification product, the migratory locust Rab5 gene is obtained, and the sequence is shown as SEQ ID NO: 1. Furthermore, according to SEQ ID NO: 1, a corresponding primer is designed and synthesized, a gene segment with the nucleotide sequence shown in SEQ ID NO: 2 is obtained through PCR amplification, the gene segment is subjected to in-vitro transcription according to a kit specification to synthesize dsRNA, the dsRNA is injected into a migratory locust body cavity to specifically silence a target gene, migratory locust dies during ecdysis, and multiple experiments show that the fatality rate reaches 100%. Due to the specificity and the efficient fatality rate of the dsRNA, the dsRNA has important practical significance for migratory locust prevention and control, and a new way can be provided for migratory locust prevention and control.

The invention belongs to the field of cell biology and immunology, and relates to a CD28 bidirectional activation co-stimulatory molecular receptor and applications thereof. In particular, the presentinvention relates to a CD28 bidirectionally activated co-stimulatory molecular receptor which successively comprises, from the N-terminal to the C-terminal, an optional signal peptide, a CD28 activated single chain antibody, an extracellular hinge region, a transmembrane region and an intracellular co-stimulatory signal molecule. When the CD28 bidirectionally activated co-stimulatory molecular receptor modies T cells together with the first-generation of CAR-T containing the first signal, a strong clustering effect can be produced, and the tumor cells can be killed. Meanwhile, the bidirectional activation effect is only limited between the T cells in contact with each other, which is not like the CD28-injected activated antibody which triggers strong T cell immunity and causes potential serious toxic and side effects.

The invention provides an antibody. The antibody comprises a protein dimer, wherein the protein dimer comprises a first functional domain including an immunoglobulin chain or an antibody combining site of a specifically-recognized specific antigen and a second functional domain including an immunoglobulin chain or an antibody combining site of a specifically-recognized specific antigen. The antibody is characterized in that the first functional domain consists of a first VH structural domain and a first VL structural domain, and the second functional domain consists of a second VH structural domain. The VH is a heavy chain variable zone, and the VL is a light chain variable zone. The bifunctional antibody provided by the invention has the advantages that a half-life period is greatly prolonged; and a homodimer is extremely less, so that identification and removal can be carried out only through primary purification even if aggregates occur.

The invention relates to colon targeting recombinant toxin as well as a preparation method and application thereof, and belongs to the colon targeting recombinant toxin prepared by using genetic engineering. The colon targeting recombinant toxin has an amino acid sequence as shown in SEQID No.2, pET28a is used as an expression vector, and BL21(DE3) is used as an expression host, so that a BL21(DE3)(pET28a-rCCK96-104PE38KDEL) recombinant engineering bacterial strain is constructed; and 6-poly-L-histidine labels are carried, so that purification is facilitated. A Ni-NTA metal-chelating affinity column is used, and a specificity cutting method for rTEV enzymes is combined, so that only through two-step purification, rCCK96-104PE38KDEL recombination protein being high in activity, high in purity and free from redundant sequences can be obtained, and the purity is 95% or above; in vitro cell experiment, purified rCCK96-104PE38KDEL recombinant immunotoxin can specifically kill colon cancer cells with high expression, can restrain proliferation and diffusion of cancer cells, and does not have any damaging effect on normal cells and other non-CCK2R high-expression tumor cells; and therefore, the rCCK96-104PE38KDEL conforms to conditions as tumor targeting drugs, and a new direction is provided for treatment of the colon cancers.

The invention belongs to the fields of cell biology and immunology, and relates to a CD137 bidirectional activation costimulatory molecule receptor and usage thereof. Specifically, the invention relates to a CD137 bidirectional activation costimulatory molecule receptor which sequentially comprises, from N terminal to C terminal, the following elements: optional signal peptides, CD137 activated single-chain antibodies, an extracellular hinge region, a transmembrane region, and intracellular costimulatory signaling molecules. When the CD137 bidirectional activation costimulatory molecule receptor modifies T cells together with a first-generation CAR-T containing a first signal, a strong cluster effect can be produced so as to kill tumor cells. Furthermore, the bidirectional activation effect is limited only between T cells which are in contact with each other, so that, strong T cell immunity is not triggered as it does when activated antibodies to CD137 are injected; and thus, potentialserious side effects are avoided.

The invention discloses a preparation method of a protozoa cyst transmission electron microscope sample, which is characterized in that a traditional fixing method is improved, and glutaraldehyde and Tritan X-100 are used for treating a cyst under the condition of not damaging the cyst, the glutaraldehyde plays a fixing role, and Tritan X-100 has the characteristic of increasing the permeability of a eukaryotic cell membrane, has no killing effect on microorganisms, and can promote a stationary liquid to penetrate through a cyst wall, so that a sample is quickly fixed, and the difficulty that the sample is not fully fixed in the past is solved. An Epon812 epoxy resin embedding agent in a traditional method is replaced by the Spurr embedding agent, so that the embedding agent can better and completely permeate into the cyst. According to the method, the treatment conditions in each step are optimized according to the characteristics of the cyst, so that the effectiveness of a sample preparation result is improved, and a basis is provided for clearly presenting the sample. The method can be applied to preparation of most protozoa cyst TEM samples, and has certain stability and repeatability.

The invention relates to a preparation method of a magnetic nano photosensitizer composite system. The method includes preparing Fe3O4 magnetic nano particles; coating a thin layer of silicon dioxideon the surfaces of the Fe3O4 magnetic nano particles by using ethyl orthosilicate as a modifier; and assembling quantum dots CdTe and a photosensitizer on the surfaces by utilizing the adsorption effect of silicon dioxide boundary pore microspheres to obtain magnetic nano photosensitizer particles Fe3O4@SiO2@Qdots / PS. The method can utilize tumor cell specific bioluminescence as a light source, the photosensitizer is excited to generate active oxygen clusters, and electric energy can be converted into heat energy in an alternating electric field, so that the magnetic nano photosensitizer is anovel multifunctional photosensitizer.

The invention belongs to the fields of cell biology and immunology, and relates to a bidirectional activated costimulatory molecule acceptor and use thereof, in particular to a bidirectional activatedcostimulatory molecule acceptor. The bidirectional activated costimulatory molecule acceptor sequentially comprises the following elements from the N terminal to the C terminal: selected signal peptide, a costimulatory signal molecule activated single-chain antibody, an extracellular hinge region, a transmembrane region and an intracellular costimulatory signal molecule. When the bidirectional activated costimulatory molecule acceptor and first generation CAR-T comprising the first signal jointly modify T cells, strong colony effect can be generated, and tumor cells can be killed. Besides, the bidirectional activating effect is only limited between the T cells in mutual contact, and cannot cause strong T cellular immunity so as to cause serious toxic and side effects like an activated antibody of injection costimulatory signal molecules.

The invention relates to a recombinant adeno-associated virus AAV-shCdc6 formulation, which contains the recombinant adeno-associated virus AAV-shCdc6 which is obtained by inserting the small interfering RNA (siRNA) of a specific human Cdc6RNAi targeting sequence to the special site of the AAV (Adeno-Associated Virus). The recombinant adeno-associated virus AAV-shCdc6 formulation provided by the invention is capable of efficiently infecting the breast cancer cells and continuously expressing the siRNA of the specific targeting Cdc6, and finally capable of specifically killing the breast cancer cells; on the contrary, the recombinant adeno-associated virus AAV-shCdc6 formulation has no killing effect on normal mammary epithelial cells. Therefore, the recombinant adeno-associated virus AAV-shCdc6 formulation can be used as a novel biological formulation for treating the breast cancer.

The invention discloses a fully human anti-MAGEA1 all-molecule IgG antibody. The fully human anti-MAGEA1 all-molecule IgG antibody comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the light chain variable region is as shown in SEQ ID NO. 3, or the sequence is a conservative variant obtained by conservative mutation added, deleted, substituted and modified by one or more amino acids; and the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO. 4, or the sequence is a conservative variant obtained by conservative mutation added, deleted, substituted and modified by one or more amino acids. The invention further discloses DNA molecules, an expression vector, host cells and application of the all-molecule IgG antibody.

The invention discloses target specific double mutant fusion protein, which is established by human gonadotropin releasing hormone mutant (mGnRH) and recombinant pseudomonas aeruginosa exotoxin A mutant (PE38m4a) fusion protein. The mGnRH-PE38m4a protein provided by the invention has obvious specific binding activity and cytotoxicity to the tumor cell lines which comprise colon cancer HT-29 cells, oophoroma OVCAR3 cells, cervical adenocarcinoma HeLa cells or liver cancer HepG-2 cells, and the like; the normal cells basically have no lethal function; the activity of the fusion protein is enhanced by approximate nine to ten times compared with the activity of natural PEA; the cost is low, the method is simple, therefore the target specific double mutant fusion protein is more suitable for industrialized production.

The invention relates to a multiple antigen peptide (MAP) vaccine and application thereof. The peptide sequence of the MAP vaccine is shown as SEQ ID NO:1-3. The MAP vaccine is a human telomerase reverse transcriptase (hTERT)-based T cell epitope MAP vaccine. The peptides shown as the SEQ ID NO:1-3 are selected from HLA-A2 restrictive dominant epitopes P540 and P865 and recessive epitope distinct peptide P572Y. The invention also provides an anti-tumor preparation. The anti-tumor preparation comprises the MAP vaccine. The MAP vaccine provided by the invention has obvious anti-tumor activity and does not have killing effect on tumor cells only expressing HLA-A2+ or hTERT+. A cytotoxic T lymphocyte (CTL) reaction induced by the MAP vaccine does not have any killing side effect on autologous lymphocytes and dendritic cells (DC), so the MAP vaccine is safe to use. The number of specificity T lymphocytes activated by the MAP vaccine is increased obviously.

The invention relates to mulberry endophytic fungi podosporia sp. 7GJ-4. The strain is collected in the China Center for Type Culture Collection on March 24, 2018 and has a collection number of CCTCCNO:M 2018118. The strain has the following advantages that endophytic fungi with the strong antagonistic effect on pathogenic bacteria of mulberry constrictive sclerotinios, namely mulberry scleromitrula shiraiana, are obtained by separation from a mulberry tree, and morphological and molecular biological identification proves that the strain is a new species of podosporia; the strain can grow within the temperature range of 15-30 DEG C, sterile fermentation filtrate has relatively high thermal stability, and the strain can adapt to the field temperature and has no lethal effect on mulberry tissue culture seedlings; and therefore, the strain has potential commercial development and application values, and has important application prospects in biological prevention and control practice ofmulberry sclerotinia.

The invention discloses Sansevieria trifasciata polysaccharides as well as a preparation method and application of the Sansevieria trifasciata polysaccharides. The Sansevieria trifasciata polysaccharides are prepared by the steps of: drying Sansevieria trifasciata leaf, crushing, screening, adding a certain volume of petroleum ether, refluxing with ethanol to remove impurities, carrying out suction filtering on the extract to obtain filter residue, and airing the filter residue for standby; extracting polysaccharides of Sansevieria trifasciata by a hot water leaching method, deproteinizing crude polysaccharides by combining an enzyme method with a Sevag method, and separating and purifying the deproteinized Sansevieria trifasciata polysaccharides TPS is by DEAE-52 column chromatography and SephdexG-100 column chromatography to respectively obtain four uniform components: TPS-A, TPS-B, TPS-C and TPS-H. TPS, TPS-H and TPS-A have obvious inhibitory action on the growth of human lung cancer cell A549, human nasopharyngeal carcinoma cell line CNE and human skin T cell lymphomata Hut102, and TPS-A has the best effect; and TPS, TPS-H and TPS-A have no inhibitory action on human colon cancer cell LoVo cell, and have no lethal effect on human intact liver cell LO2, thereby providing scientific reference for research and development of targeted antitumor medicaments.

The invention belongs to the field of cytobiology and immunology and relates to a CD40 dual costimulatory activated receptor (DCR) and an application thereof. Particularly, the CD40 DCR sequentially comprises the following elements from N end to C end: optional signal peptide, a CD40 activated single-chain antibody, an extracellular hinge region, a transmembrane region and an intracellular costimulatory signal molecule. The CD40 DCR and first-generation CAR-T containing a first signal can produce a strong cluster effect and kill tumor cells when co-modifying T cells. meanwhile, the dual activation action is only limited among contacted T cells and cannot cause strong T cell immunity and cause potential severe toxic and side effects like injection of a CD40 activated antibody.

The invention discloses antigen epitope peptide. An amino acid sequence contains an amino acid sequence shown in SEQ ID No.2 or SEQ ID No.3, or an amino acid sequence which is formed by adding, deleting or replacing one or multiple amino acids of the amino acid sequence shown in SEQ ID No.2 or SEQ ID No.3. The antigen epitope peptide can be used for producing antigen presenting cells, main tissue compatibility antigen compounds or cell toxic T lymphocyte inducing agents, and can be applied to prepare medicines for treating cancers, such as glioma.

A recombinant adenovirus configurator deleted 19kDa E1B (1713-2243nt) and E1B 55KDa (2018-3328 nt) sequence in human adenovirus type 5 genom and its application to diagnosing and treating tumor are disclosed. The PCR amplification, enzyme severing, linking, subcloning, transfection, and single-cloning purification of recombinant adenovirus technologies, and the deleted 19kDa E1B coded area (1713-2243 nt) and E1B 55kDa (2018-3328 nt) sequence are used to screen the said comfigurator. It can selectively kill tumor cells, but not normal cells.