53 results about "Cytopathic effect" 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 provides a porcine epidemic diarrhea virus G2c subtype strain and application thereof. According to the invention, the porcine epidemic diarrhea virus G2c subtype HN2501 strain is separated from porcine intestinal tissues and is subjected to passage purification, and the microbial preservation number of the porcine epidemic diarrhea virus G2c subtype HN2501 strain is CGMCC (China General Microbiological Culture Collection Center) No.46417. The isolated strain can stably proliferate on passage cells to generate typical cytopathy. The G2c subtype isolated strain of the porcine epidemic diarrhea virus has excellent immunogenicity; the vaccine prepared from the isolated strain can induce piglets to generate high-level neutralizing antibodies; meanwhile, the ELISA kit prepared from the isolated strain can be used for well detecting the porcine epidemic diarrhea virus antibody in serum.

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 provides an oligopeptide with a fish virus resisting effect, and the oligopeptide is used as a feed additive or a liquid protein preparation for injection to kill viruses or inhibit the proliferation of the viruses, so that the treatment of the fish virus diseases is realized. The amino acid sequence of the oligopeptide is SEQ ID NO: 1. The Pt5-1c oligopeptide provided by the invention reduces the cytopathic effect degree of GCRV virus infection on EPC, and shows that the Pt5-1c oligopeptide has an important antiviral effect. By adding the Pt5-1c oligopeptide into the feed or injecting the Pt5-1c oligopeptide into the fish, the purpose of preventing and treating viral diseases of the fish can be achieved.

The present invention relates to the field of biotechnology. The present invention provides a grass carp brain cell line sensitive to grass carp reovirus type II, which was deposited in the China Center for Type Culture Collection on October 19, 2024, with a deposit number of CCTCC NO: C2024101. The grass carp brain cell line constructed by the present invention shows remarkable stability and successfully achieved 60 stable passages. When inoculated with GCRV‑II, an atypical cytopathic effect can be exhibited. In addition, the grass carp brain cell line has the ability to efficiently replicate GCRV‑II, and even after three generations of blind transmission, the generated cytotoxin can still cause grass carp to become ill and die. Therefore, the present invention has laid a solid foundation for in-depth exploration of the pathogenic mechanism of GCRV‑II, vaccine development, and antiviral drug screening, which will help promote further progress in related research and provide a key cell model and research basis for the formulation of grass carp reovirus-related research and prevention and control strategies.

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 present invention relates to an aryl sulfonamide compound and its anti-influenza use, belonging to the technical field of biomedicine. The present invention provides a series of aryl sulfonamide compounds represented by the general formula I. Experimental verification shows that the aryl sulfonamide compounds have excellent antiviral activity. The toxicity of the compounds is evaluated by cytopathic effect assay. The compounds provided by the present invention have ideal safety and good development prospects.

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 invention discloses construction and application of a recombinant PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) strain for expressing porcine circovirus Cap protein. The recombinant PRRSV capable of expressing the Cap protein of the porcine circovirus type 2 is successfully constructed and saved by adopting a reverse genetic manipulation technology. The modified virus is inoculated to Marc-145 cells for subculture, and the result shows that the modified virus can trigger cytopathy and stably express Cap protein. The obtained recombinant strain shows a stable passage capability in a Marc-145 cell, and has an extremely high virus load. The strain can be used as a vaccine candidate strain, provides multiple protection for PRRSV and porcine circovirus of different pedigree, and provides a brand new solution for healthy breeding of pigs.

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.

A viral vector delivery system for both respiratory and digestive tracts of pigs and an application thereof are provided. The viral vector delivery system includes a backbone plasmid and a helper plasmid. The backbone plasmid is produced by inserting a full-length cDNA of porcine enterovirus B (PEVB) into a pUC57 plasmid. The helper plasmid is produced by inserting a green fluorescent protein-coding gene into a plasmid pCAG-T7-polymerase. The viral vector delivery system can be constructed with high efficiency, can quickly cause a cytopathic effect (CPE) after infecting cells, has a viral titer up to 107.75 TCID50 / mL, and can maintain high stability. A pathogenic epitope for the respiratory and digestive tracts of pigs is inserted into the backbone plasmid to produce a vaccine antigen, which is non-pathogenic, has high stability and a high viral titer, and allows a prominent immunization effect after being inoculated in pigs.

This invention discloses the use of an anthraquinone natural product (Aloesaponarin II) in the treatment of enterovirus 71. Experimental studies of Aloesaponarin II's anti-EV71 activity have shown that Aloesaponarin II inhibits the cytopathic effect (CPE) produced by EV71 on host cell RD, enhances cell survival, and reduces progeny virus production, suggesting its potential application in the preparation of anti-EV71 drugs.

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.

The invention provides an application of 5-O-methylvisammioside in preparation of a medicine for inhibiting HCMV (human cytomegalovirus) infection. According to the present invention, the 5-O-methylvisammioside can significantly reduce the cytopathic effect caused by HCMV infection, and the DNA copy numbers of the immediate early protein IE1 / 2, the early protein p52, the immediate early gene UL123, the early gene UL44 and the late gene UL32 of the HCMV are selected as the detection indexes to discover that the 5-O-methylvisammioside can provide the significant inhibition effect, and finally, a medicine addition experiment finds that the 5-O-methylvisammioside can play an antiviral role in multiple stages of a virus life cycle. The 5-O-methylvisammioside has small toxic and side effects, plays an antiviral role in multiple stages, has good application prospects and development advantages, and is expected to be developed into an ideal medicine for treating and / or preventing HCMV infection.

The present invention discloses an anti-dengue virus compound and its application, belonging to the technical field of antiviral agents. The present invention can significantly inhibit the cytopathic effect induced by DENV-2 on cells, reduce the production of viral ions, inhibit the replication of DENV-2 virus, and reduce the expression of DENV-2E protein, showing significant anti-dengue virus effects.

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.

This patent discloses the use of voxtalisib in the preparation of anti-ADV7 drugs. Experimental studies of voxtalisib's anti-ADV7 activity have shown that the compound voxtalisib inhibits the cytopathic effect (CPE) produced by ADV7 in Hela cells, enhancing cell survival and reducing progeny virus production, suggesting its potential application in the preparation of anti-ADV7 drugs.

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.