30 results about "Cytopathic change" patented technology

The application discloses an isolation method of bovine epidemic fever virus and the bovine epidemic fever virus obtained by the method and application of the bovine epidemic fever virus, and belongs to the technical field of bovine epidemic fever virus isolation. The application aims to provide an efficient and rapid method for isolating bovine epidemic fever virus. The application provides the isolation method of the bovine epidemic fever virus, wherein the anticoagulated blood sample of a bovine is inoculated on KC cells for culture, the virus is harvested, the BHK-21 cells are inoculated to grow into a monolayer, and the first generation of passage cells with cytopathic effect is obtained by passage. The virus strain can also be used for screening, preparation and detection of bovine epidemic fever virus antibodies, and for preparation of an animal model infected by the bovine epidemic fever virus.

The invention belongs to the technical field of antiviral drugs, and relates to application of a pyrimidine derivative as a coxsackie virus CVB3 inhibitor. The anti-CVB3 activity research experiments of several pyrimidine derivatives show that the pyrimidine derivatives inhibit the cytopathic effect (CPE) generated by CVB3 on host cells Hep-2, enhance the cell survival rate and reduce the progeny virus yield; the inhibitory activity of the pyrimidine derivative on CVB3 shows that the pyrimidine derivative has application potential in preparation of anti-CVB3 virus drugs.

The invention discloses a bovine virus diarrhea live vaccine based on suspension culture and a preparation method thereof, and relates to the field of animal vaccines. The bovine viral diarrhea virus vaccine is a freeze-dried vaccine, and each vaccine contains a bovine viral diarrhea virus BVDV attenuated strain antigen and a freeze-drying protective agent; wherein the virus content of the antigen of the BVDV attenuated strain is 105-107 CCID50 / piece, and the BVDV attenuated strain is a cytopathic effect type attenuated strain suitable for suspension culture; the freeze-drying protective agent comprises trehalose, cane sugar, gelatin hydrolysate, sodium glutamate and polyvinylpyrrolidone. The suspension culture technology is adopted, large-scale, efficient and stable production of BVDV antigens is achieved, an optimized freeze-drying formula is combined, the obtained vaccine is high in virus titer, good in immunogenicity, high in stability, good in safety, advanced in production process, controllable in cost and suitable for industrial large-scale production.

The invention discloses an application of an ST6GALNAC5 gene in prevention and treatment of swine Graisseria disease (a pathogenic bacterium causing the swine Graisseria disease by a haemophilus parasuis system), and the expression of the ST6GALNAC5 gene is inhibited in a targeted manner, so that adhesion and invasion of the haemophilus parasuis to host cells can be effectively inhibited, and cytopathy is reduced. The method is helpful for resisting cell death induced by haemophilus parasuis infection, so as to prevent the occurrence of the porcine Graisseria disease. Therefore, the ST6GALNAC5 gene can be used as an important target spot for treating the swine Graisseria disease, and has important clinical application prospects and molecular disease-resistant breeding values.

The application provides a porcine epidemic diarrhea virus mutant strain and application thereof. The application obtains a porcine epidemic diarrhea virus mutant strain GX2506 from porcine intestinal tissue, and the strain is purified through passage, and the microorganism preservation number of the strain is CGMCC No. 47089. The isolated strain can stably proliferate on a passage cell, and typical cytopathic effect is generated. The porcine epidemic diarrhea virus mutant isolated strain has excellent immunogenicity; a vaccine prepared by using the isolated strain can induce piglets to generate high-level neutralizing antibodies; meanwhile, an ELISA kit prepared by using the isolated strain can better detect porcine epidemic diarrhea virus antibodies in serum.

Disclosed are an rBJ-VVL plasmid, a mutant strain of NADC34-like PRRSV-2 and a preparation method therefor and application thereof. Further disclosed is an NADC34-like PRRSV-2-specific vaccine. In the present disclosure, a modified strain rBJ-VVL with tropism for Marc-145 cells is obtained by precisely mutating an amino acid at positions 91 / 97 / 98 of GP2a; the modified virus constructed in the present disclosure can be propagated in Marc-145 cells, cause cytopathic effects and form plaques when inoculated into Marc-145 cells for serial passage; the resulting Marc-145 cell-passaged viruses have an extremely viral load, and a large number of new progeny viruses can be obtained in a short time; and the Marc-145-adapative modified strain cultured in the present disclosure is used to create a first NADC34-like PRRSV-2-specific vaccine.

The invention discloses an application of a compound Darifenacin hydrobromide in preparation of a medicine for resisting influenza A (H1N1) virus, and belongs to the technical field of medicines. The invention discloses for the first time that Darifenacin hydrobromide can significantly inhibit replication of H1N1-PR8 virus, reduce virus titer and reduce cytopathic effect (CPE) in an in-vitro cell model, the EC50 value of Darifenacin hydrobromide is 2.5 [mu] M, and the selection index (SI) is gt; and 10, the compound has good antiviral activity and safety. The compound Darifenacin hydrobromide is a high-selectivity M3 muscarinic acetylcholine receptor antagonist and is approved to be used for treating overactive bladder at present, the invention discloses that the compound can inhibit replication and release of H1N1-PR8 virus for the first time, and the compound has a wide prospect of being developed into a medicine for treating human influenza virus infection.

The application discloses a porcine reproductive and respiratory syndrome mutant virus and a construction method and application thereof. The application accurately analyzes key amino acid sites of PRRSV-1 determining Marc-145 cell tropism, specifically, 88(F), 94(I) and 95(L) amino acids of GP2a, 70(G), 72(N), 80(D), 154(I), 158(H) and 163(L) amino acids of GP3, and 49(D), 53(L), 54(R) and 57(G) amino acids of GP4. Three Marc-145 cell non-adapted strains are precisely point mutated, three PRRSV-1 modified strains which can adapt to Marc 145 cell in vitro passage culture are obtained, the strains can be stably passed in vitro and have good replication efficiency, and Marc-145 cells infected by the strains can produce obvious cytopathic effect, and the strains are expected to be used as a new type of PRRSV-1 vaccine.

The invention discloses a scorpion margaritae polypeptide with anti-EV71 virus activity and application thereof, and belongs to the technical field of traditional Chinese medicinal material polypeptides. The Buthus martensii Karsch polypeptide with the anti-EV71 virus activity is Pep 15 or Pep 16; the amino acid sequence of the Pep 15 is as shown in SEQ ID NO. 1; the amino acid sequence of the Pep16 is as shown in SEQ ID NO. 2. The obtained buthus martensii polypeptide has remarkable anti-EV71 virus activity, inhibits the cytopathic effect generated by EV71 virus on host cells Vero, increases the survival rate of infected cells and inhibits replication and proliferation of EV71 virus in cells, has no toxic effect on cells when the concentration of the buthus martensii polypeptide reaches 200 mu M, has the potential of being used as an anti-EV71 virus drug, and can be used as a drug for treating EV71 virus. The compound has a good application prospect in the aspect of developing a novel medicine for preventing and treating the hand-foot-and-mouth disease of infants induced by the EV71 virus.

The invention discloses application of a serine protease inhibitor in preparation of antiviral drugs, and belongs to the technical field of biological medicines. The two serine protease inhibitors, namely the camostat mesylate and the nafamostat mesylate, can almost completely inhibit cytopathy under the action of the concentration of 2 mu M, have a remarkable effect of inhibiting virus replication and proliferation in various PEDV susceptible cells, reduce the mRNA (messenger ribonucleic acid) level of PEDV N (porcine epidemic diarrhea virus N) and inhibit the expression of PEDV N protein. The compound is used for preparing the medicine for resisting the porcine epidemic diarrhea virus, and has the advantages of reliable curative effect, no toxic or side effect, abundant resources, low price and the like.

The invention provides a diaminopyrimidine compound with a structure as shown in a formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof, or a prodrug molecule thereof, or a deuteride thereof, or a tritium thereof, and an application of the diaminopyrimidine compound, or the stereoisomer thereof, or the pharmaceutically acceptable salt thereof, or the solvate thereof, or the prodrug molecule thereof, or the deuteride thereof, or the tritium thereof. The diaminopyrimidine compound provided by the invention has obvious inhibitory activity on RSV virus, has a chemical structure type completely different from that of the existing anti-RSV drug ribavirin and the like, is expected to be developed into a novel anti-RSV drug, is used for inhibiting replication or reproduction of the RSV virus, and has broad application prospects. The pharmaceutical composition can be used for preventing, relieving and / or treating respiratory syncytial virus infection and cytopathic effects caused by the respiratory syncytial virus infection, and can be used for treating bronchitis or pneumonia and other diseases caused by the respiratory syncytial virus infection.

The invention discloses an application of MG-132 in preparation of a medicine for resisting nervous necrosis virus. Researches show that the MG-132 can significantly reduce cytopathic effects caused by nervous necrosis virus infection, reduce the number of positive cells expressing RGNNV capsid proteins, and also can significantly reduce the mRNA transcriptional level and protein expression level of RGNNV key genes and the copy number of viral genome RNA; and along with the prolonging of RGNNV infection time, the MG-132 can also significantly inhibit virus gene transcription, protein synthesis and genome replication, and reduce virus titer. Therefore, the virulence of RGNNV on cells can be remarkably reduced through MG-132 treatment, the effect of resisting nervous necrosis virus is achieved, the cytotoxicity is low, and the safety is good. The invention provides a new drug intervention strategy for prevention and treatment of nervous necrosis virus infection, and has important application value for prevention and treatment of NNV related diseases in aquaculture.

The application discloses a method for determining baculovirus titer, which adopts a mouse IgG2a type monoclonal antibody specifically recognizing envelope protein gp64, and comprises the following steps: step 1, cell plating: cells are plated in a cell culture plate; step 2, baculovirus inoculation: a baculovirus sample to be detected is inoculated into the cell culture plate in step 1; step 3, virus infection; and step 4, flow cytometry detection: infected cells are collected, excessive baculovirus is removed, the cells are washed, fluorescently labeled antibodies are added, and machine detection is carried out. The method of the application is more simple in operation process by using the to-be-detected baculovirus to directly infect insect cells and flow cytometry analysis based on the gp64 antibody, and does not depend on subjective judgment of an experimenter on cytopathic effect, so that the result is more representative.

The invention provides application of levistilide A in preparation of anti-influenza virus drugs, and belongs to the technical field of biological medicines. The influenza virus is an H1N1 virus. In-vitro anti-influenza experiments find that levistilide A can significantly inhibit infection of H1N1 virus and significantly improve cytopathy caused by influenza virus infection. The H1N1 virus half inhibitory concentration (IC50) of the levistilide A in MDCK cells is 1.462 M, and the H1N1 virus half inhibitory concentration (IC50) of the levistilide A in A549 cells is 0.455 M. The invention proves that the levistilactone A has a remarkable anti-H1NA virus effect, can be used for preparing the anti-influenza virus medicine, and has an excellent application prospect.

The invention relates to a cat omega interferon biological activity detection method and a kit. According to the method, a stable reporter cell line is established by constructing a recombinant plasmid containing a cat omega-interferon specific interferon stimulus response element (ISRE) and a reporter gene (such as luciferase) and transfecting CRFK cells. During detection, report cells and a sample to be detected are co-incubated, cat omega-interferon induces report gene expression by activating a JAK-STAT-ISRE signal channel, the expression level is in direct proportion to interferon activity, and the activity can be quantified by detecting a luminescence value (RLU). The method overcomes the defects of long period (3-5 days), complicated operation and subjective result judgment of the traditional cytopathic effect inhibition method, has the advantages of rapidness (about 24 hours), safety (no live virus), objectiveness, accuracy, good repeatability and easiness in high throughput, and is suitable for quality control analysis of cat omega-interferon products.

The invention discloses an application of a phthalocyanine compound and a delivery system thereof in preparation of an anti-clostridium difficile toxin drug, and discovers that phthalocyanine can specifically target TFPI, CSPG4 and FZD1, 2, 7 receptor binding domains of clostridium difficile toxin B (TcdB), and effectively inhibits binding of toxin and host cells. In-vitro experiments prove that the phthalocyanine (Phthalocyanine, Pc) can be used for remarkably relieving the cytopathy induced by TcdB and has no cytotoxicity. In order to improve the curative effect, an extracellular vesicle (S-EV) from saccharomyces boulardii is creatively adopted to construct a drug loading system S-EV-Pc, and the pH responsive intestinal targeted delivery of the drug is realized. Animal experiments show that the preparation can significantly improve the survival rate of infected gerbil to 100%, significantly improve diarrhea symptoms, reduce intestinal TcdB load and relieve tissue pathological damage. The invention discloses the potential of phthalocyanine as a non-antibiotic small molecule drug in treatment of clostridium difficile infection (CDI) for the first time, and provides a brand new strategy and an experimental basis for developing a novel anti-CDI preparation.

This invention discloses the application of nano-zinc oxide in the preparation of drugs against fish iridovirus (SGIV). The study shows that nano-zinc oxide (ZnONPs) can significantly attenuate the cytopathic effect induced by SGIV infection, reducing the mRNA transcription level, protein expression level, and viral titer of the major SGIV capsid protein. Treatment with ZnONPs can also enhance the host's interferon immune-related genes ISG56 and ISG20, and anti-inflammatory factors IL-10 and TGF-β. β The ZnONPs significantly reduced the mRNA transcription level, thereby decreasing the infectivity of SGIV to cells and demonstrating antiviral activity against fish iridoviruses. Furthermore, ZnONPs exhibited low cytotoxicity and good safety profile, making them suitable for the preparation of drugs to prevent and treat grouper iridovirus disease. This invention provides a novel drug intervention strategy for the prevention and treatment of SGIV infection and has significant application value in the control of iridovirus-related diseases in aquaculture.

The application discloses application of MG-132 in preparation of a drug for resisting nerve necrosis virus. Researches of the application show that MG-132 can significantly reduce the cytopathic effect caused by nerve necrosis virus infection, reduce the number of positive cells expressing RGNNV capsid protein, and also can significantly reduce the mRNA transcription level of a key gene of RGNNV, the protein expression level and the copy number of virus genomic RNA; and with the extension of RGNNV infection time, MG-132 can also significantly inhibit virus gene transcription, protein synthesis and genome replication, and reduce virus titer. Therefore, MG-132 treatment can significantly reduce the virulence of RGNNV to cells, has the efficacy of resisting nerve necrosis virus, and has low cytotoxicity and good safety. The application provides a new drug intervention strategy for prevention and treatment of nerve necrosis virus infection, and has important application value for prevention and control of NNV related diseases in aquaculture.

The application discloses a construction method of Aedes albopictus Aa01 cell line and susceptible cell model of Decapod iridovirus 1, and belongs to the technical field of virology and aquaculture disease prevention and control, wherein the Aedes albopictus Aa01 cell line is preserved in the China Center for Type Culture Collection of Wuhan University, and the preservation number is CCTCC NO: C202648. The method for constructing the DIV1 susceptible cell model based on the above cell line comprises the following steps: culturing the Aedes albopictus Aa01 cell line, and subculturing or using for virus inoculation when the density reaches 80%-90%; inoculating the DIV1 into the Aedes albopictus Aa01 cell line, removing the virus liquid after virus adsorption, and adding cell maintenance liquid for continuous culture; and observing the cytopathic effect CPE, collecting the cell culture when the CPE reaches 70%-80%, and obtaining the amplified DIV1 virus liquid. The DIV1 susceptible cell model constructed by the method can solve the problem that there is currently a lack of stable and continuously subcultured DIV1 in vitro culture.

The application discloses a weak strain of Cyprinid herpesvirus 2 (CyHV-2) G-RP7 for preparing a common carp hematopoietic organ necrosis disease vaccine, and the preservation number is CGMCC No. 24099. The weak strain is used to infect a goldfish fin cell line (RyuF-2), and after 4-5 days, typical cytopathic effect (CPE) is observed on the cells, and the continuous passage can stably cause the cytopathic effect. The weak strain is sensitive to the RyuF-2 cell line, realizes in-vitro stable culture and amplification of the weak strain, and the TCID50 of the virus suspension is determined by a Reed-Muench method 50 up to 10 5.9 / ml. The virulence of the weak strain does not return to be strong after continuous in-vitro and in-vivo passages. The weak strain can be used for preparing the common carp hematopoietic organ necrosis disease vaccine, and both immersion immunization and intraperitoneal injection immunization can stimulate the fish to produce specific immune response, and have good anti-CyHV-2 infection effect.

The invention relates to the technical field of virus construction, and discloses an Acaban virus rescue construction method and application thereof, and the method comprises the following steps: designing primers aiming at three segments S, M and L of an Acaban virus genome, adding a T7 promoter sequence at the 5'terminal of an upstream primer, and adding a T7 terminator sequence at the 5 'terminal of a downstream primer; carrying out amplification on each gene segment of the Acaban virus by utilizing the primers; carrying out enzyme digestion on the plasmid containing the T7 promoter and terminator sequence to obtain a linearized plasmid; carrying out homologous recombination on the PCR product and the linearized plasmid to construct three recombinant plasmids; uniformly mixing the three recombinant plasmids, and co-transfecting cells for expressing T7 polymerase; culturing and collecting supernatant, inoculating BHK-21 cells, and carrying out continuous passage until cytopathic effect occurs, so as to obtain the rescued acarban virus. According to the invention, the accurate transformation of the viral genome is realized, and the virus rescue efficiency and the vaccine safety are improved.

The application discloses a porcine epidemic diarrhea virus G2b subtype strain and application thereof. The application obtains a porcine epidemic diarrhea virus G2b subtype GZ2504 strain from a porcine intestinal tissue, and the strain is purified through passage, and the microorganism preservation number of the strain is CGMCC No. 47083. The isolated strain can stably proliferate on a passage cell and produce a typical cytopathic effect. The porcine epidemic diarrhea virus G2b subtype isolated strain has excellent immunogenicity; a vaccine prepared by using the isolated strain can induce piglets to produce high-level neutralizing antibodies; meanwhile, an ELISA kit prepared by using the isolated strain can better detect porcine epidemic diarrhea virus antibodies in serum.

The invention discloses a method for preparing an rHP-PRRSV-NADC30 Nsp230 + 131 recombinant strain and an application of the rHP-PRRSV-NADC30 Nsp230 + 131 recombinant strain. According to the present invention, by using the reverse genetic manipulation technology, the virus strain is successfully constructed and saved, and the inherent 30 deletion of HP-PRRSV (line 8.7) and the 131 deletion of NADC30-like (line 1.8) are simultaneously retained on the PRRSV Nsp2 protein of the virus strain; the modified virus is inoculated to Marc-145 cells for subculture, and experimental results show that the modified virus can induce cytopathy. The obtained recombinant strain shows a stable passage capability in a Marc-145 cell. In addition, clinical animal experiments further verify the safety and effectiveness of the recombinant strain. No obvious pathological reaction and abnormal clinical manifestation are observed in an experimental animal inoculated with the recombinant strain, which indicates that the recombinant strain has good biological safety. Meanwhile, experimental animals show that the recombinant strain can stimulate effective immune response, an effective immune protection effect is provided for multilineage PRRSV infection, and potential vaccine development prospects are shown.

The invention discloses application of ammonium ferric citrate in preparation of a medicine for resisting grouper iridovirus. The effective concentration of the ammonium ferric citrate at the cellular level is 0.04-0.08 mM, and the effective concentration of the ammonium ferric citrate in vivo is 20 mg / kg. The in-vivo and in-vitro treatment of the ferric ammonium citrate can inhibit the replication of the SGIV. The ferric ammonium citrate with a safe concentration in in-vitro cells can significantly reduce cytopathy induced by SGIV infection and reduce viral protein expression and yield of progeny viruses, intraperitoneal injection of ferric ammonium citrate can inhibit transcription expression of viral genes in vivo, reduce the death rate of groupers caused by SGIV infection, and improve the safety of the groupers. The ferric ammonium citrate has a remarkable antiviral effect and has the potential of researching and developing the anti-iridovirus medicine. In addition, the ferric ammonium citrate can improve the oxidation resistance of cells or tissues, so that tissue pathological damage caused by virus infection is improved. As an antiviral drug, the ammonium ferric citrate has a certain application value in prevention and control of grouper iridovirus diseases.

The invention discloses a porcine delta coronavirus isolate, a polyclonal antibody and application of the porcine delta coronavirus isolate, and belongs to the technical field of animal virology and biology. The porcine delta coronavirus isolate is a porcine Delta coronavirus JS strain, the preservation number is CCTCC (China Center for Type Culture Collection) NO: V202157, the preservation date is August 10, 2021, and the preservation unit is China Center for Type Culture Collection. The PDCoV JS isolate provided by the invention can be stably proliferated on LLC-PK1 cells to generate typical cytopathy, has high virus titer and excellent immunogenicity, can generate a high-titer polyclonal antibody after immunizing a mouse, can induce the mouse to generate a high-level neutralizing antibody, can be used for preventing and treating porcine delta coronavirus infection, and can be used for preventing and treating porcine delta coronavirus infection. The method has good application value, and provides an important biological material for effective prevention and control of the porcine delta coronavirus disease.

The application discloses application of Rottlerin in preparation of an anti-enterovirus drug and belongs to the technical field of biological medicines. The application first discovers that Rottlerin has strong anti-EV71 virus activity, reduces the infection rate of EV71 virus, reduces the cytopathic effect caused by EV71 virus, enhances the cell survival rate of EV71 virus infection, inhibits the replication and proliferation of EV71 virus in cells, reduces the synthesis of EV71 virus protein in cells, and thus plays the high-efficiency anti-EV71 virus activity, and the Rottlerin provides a new thought and scheme for development and clinical treatment of the anti-enterovirus drug, and therefore, the Rottlerin has a good application prospect.

Impedance-based determination of metrics derived from Cytopathic Effects (CPE), such as Tissue Culture Infections Dose (TCID) values, is provided via determining a first plurality of cellular impedances (CI) corresponding to a plurality of cell samples while performing the impedance assay; inoculating the plurality of cell samples with a plurality of different dilutions of the virus such that a first and second subsets of the plurality of cell samples are respectively inoculated with first and second dilutions of a virus; determining, after inoculating the plurality of cell samples, a second plurality of CIs; determining, from the first plurality of CIs and the second plurality of CIs, cytopathic effects (CPE) experienced by the plurality of cell samples; calculating, a CPE-derived metric.

The invention belongs to the technical field of antiviral drugs, and relates to preparation of a 2, 4, 5-polysubstituted pyrimidine derivative and application of the 2, 4, 5-polysubstituted pyrimidine derivative in resisting enterovirus 71. The anti-EV71 activity research experiment of the pyrimidine derivative shows that the pyrimidine derivative has certain inhibitory activity on EV71: the pyrimidine derivative inhibits the cytopathic effect (CPE) generated by EV71 on a host cell Vero, enhances the cell survival rate and reduces the progeny virus yield; host cell lesions caused by EV71 infection can be inhibited, and the method mainly acts on a replication and proliferation stage of EV71 in host cells; therefore, the novel pyrimidine derivatives have the potential of preparing the anti-EV71 virus medicine.

The application discloses application of icariside II in the technical field of traditional Chinese medicine pharmacy in preparation of a medicine for resisting coxsackieviruses group B3 (CVB3). The present application takes VERO (African green monkey kidney) and Hela (HeLa) cells infected by CVB3 as research models, detects the influence of icariside II on cell cytopathic effect (CPE) after infection by CVB3, on virus titer in supernatant, and on transcription level of CVB3 mRNA, and the results show that icariside II has obvious anti-CVB3 effect and can be used for preparation of a medicine for resisting CVB3.

The present application relates to a kind of compound shown in formula I or its pharmaceutically acceptable salt and its use, belong to the field of medicine.The compound or its pharmaceutically acceptable salt is high to the inhibition activity of SARS-Cov-2's Mpro main protease, low toxicity, good metabolic stability.The compound or its pharmaceutically acceptable salt can be used for the purpose of preparing the drug for inhibiting the Mpro main protease of SARS-Cov-2, and for the purpose of being used in the product for preventing, treating or reducing the replication or reproduction of coronavirus infection or its homologous variant virus and the cytopathic effect generated by it.