232 results about "Cytopathic change" patented technology

The present disclosure relates to novel compounds of formulas (1) through (19) and to a method for treating humans infected with a virus including various respiratory viruses such as members of the Paramyxoviridae family (respiratory syncytial virus (RSV), human metapneumovirus (HMPV), human parainfluenza virus (HPIV), measles virus, and mumps virus) with a compound of formulas (1) through (19). The present disclosure also relates to a cytopathic effect (CPE)-based assay that will assess virus-induced CPE for screening of compounds for treating viral diseases or inhibiting a virus.

A mutant herpesvirus that can be used a recombinant virus vector includes (a) a mutation such that the mutant virus has a reduced ability in comparison with a parent type to cause lysis of an infected cell, and (b) an inactivating mutation in a gene essential for the production of infectious virus. An example is a HSV1 mutant lacking the essential glycoprotein gH gene and having a mutation impairing the function of the gene product VP16. A heterologous gene can be carried at the site of the inactivated essential gene, e.g. a gene suitable for administering gene therapy. The vector has an increased margin of safety over known herpesvirus vectors in respect of incidence of cytopathic effects and/or risk of infection.

The invention provides a method for preparing triple vaccine for preventing porcine transmissible gastroenteritis, porcine epidemic diarrhea and porcine rotavirus. The method comprises the following steps: inoculating a host-cell line with a 90 percent grown monostratum against a porcine transmissible gastroenteritis virus, a porcine epidemic diarrhea virus and a porcine rotavirus respectively, and adding a cell maintenance media into the host-cell lines respectively to be cultured at 37 DEG C; after cytopathic effect reaches over 75 percent, collecting viruses to be stored at 20 DEG C below zero for standby; mixing the viruses according to 10 TCID50 in 1:1:1, and simultaneously adding Freund's complete adjuvant and immunopotentiator into the mixture to inactivate the mixture by formaldehyde at 37 DEG C for 24 hours; and adding an oil adjuvant into the mixture to prepare a vaccine of water-oil-water preparation. The method can be used for preparing the triple vaccine for preventing the porcine transmissible gastroenteritis, the porcine epidemic diarrhea and the porcine rotavirus so as to solve the problem that the diseases do not have an effective medicine to treat currently.

Self-propagating, fusogenic blebs are produced from cells infected with a population of Venezuelan Equine Encephalitis virus replicon particles (VRP). The self-propagating, fusogenic nature of the blebs is derived from expression of heterologous genes encoding viral fusion proteins that are incorporated into the replication defective replicon particles. The resulting blebs can be harvested from supernatants of cells displaying severe cytopathic effects. The blebs are used to make immunogenic compositions and devise methods of immunizing mammals against paramyxoviruses such as parainfluenza virus type 3.

The invention relates to a method for industrially producing a pseudorabies vaccine by using a bioreactor. The method comprises the following steps of: (1) sterilizing a microcarrier and the bioreactor, inoculating cells for vaccine production, culturing the cells, and after the cells on the microcarrier form a compact monolayer, inoculating pseudorabies virus, and continuously culturing to allow the virus to reproduce; (2) after more than 80 percent of cells are subjected to a cytopathic effect, stopping culturing, and obtaining virus liquid to prepare the pseudorabies vaccine. In the method, the pseudorabies vaccine is produced by culturing the cells in high density by the bioreactor microcarrier culture technology; and compared with the conventional roller bottle production process, the invention has the advantages that: the method has high automation control degree, saves manpower, reduces cost and is small in production land, the production can be monitored in real time, the scale is easy to expand, the produced virus has high titer and small difference among batches, and the product has stable quality and small side effect.

Methods and compositions containing a phorbol ester or a derivative of a phorbol ester are provided for the treatment of cytopathic diseases. Cytopathic diseases may be caused by a variety means suchas viral infections like HIV and AIDS, or the development of neoplasms in a mammalian subject. The methods and compositions of the invention are effective for inhibiting de novo HIV infection, upregulating viral expression from latent provirus, inhibiting HIV-induced cytopathic effects, down regulating the HIV receptor, increasing ThI cytokine expression, decreasing Th2 cytokine expression, increasing ERK phosphorylation, inducing apoptosis in malignant cells, inducing remission, maintaining remission, as chemotherapeutic agents, as well as for decreasing symptoms of cytopathic diseases and opportunistic infections that may accompany such diseasesl. Additional compositions and methods are provided which employ a phorbol ester or derivative compound in combination with at least one additional agent such as those used in HAART protocols, therapeutic agents used to treat opportunistic infections due to HIV, or chemotherapeutic agents to yield more effective treatment tools against cytopathic diseases in mammalian subjects.

The invention discloses two types of ScFv (Single Chain Variable Fragment) antibodies, encoding genes of the antibodies and applications of the antibodies for preparing the preparations for treating or preventing infectious bursal disease of chicken. Two single-chain antibodies (namely, the ScFv antibodies) are provided; and the ScFv antibodies can be specially bound with the protein 2 (VP2) (Virus Protein 2) in an IBDV (Infectious Bursal Disease Virus) structure and a plurality of IBDV strains to block the cytopathic effect (CPE) of the chicken embryo fibroblast by the IBDV so as to protect the young chicken infected with IBDV. The immune serum and egg yolk antibody are tedious in preparation, high in production cost, unstable in effect, difficult to control industrialized production quality, capable of causing horizontal disease spread and the like in a use process; the ScFv antibodies disclosed by the invention can overcome the above disadvantages and have the advantages of strong specificity, good treatment effect, controllable industrialized production quality, capability of preventing the horizontal disease spread caused by the egg yolk antibody and the like; and therefore, by virtue of the ScFv antibodies, a new situation is opened up in the IBDV prevention and treatment history, and even in the entire prevention and treatment history for animal virus diseases.

The invention belongs to the field of antiviral drugs. The invention provides applications of Gramine and derivatives thereof to preparation of medicaments for resisting adenovirus Type 7. Gramine derivatives are compounds shown in G1,G2,G3,G4,G5,G10,G17,G18,G19,G20, and G21. Experiments for researching activity of Gramine derivatives for resisting ADV7, pharmacology research of Gramine derivatives for resisting ADV7, and tests of inhibition effects of G19 for ADV7 duplication show that Gramine derivatives can inhibit cytopathic effects (CPE) generated by ADV7 on host cell Hela, enhance cell survival rate, inhibit yield of progeny virus, and mainly inhibit the duplication phase of ADV7 virus in host cells. The Gramine and derivatives thereof have potential for preparing anti-ADV1 virus medicaments, and the compounds have the advantages of simple synthesis process, and economy and fastness; the compounds are easy for large-scale production, and have clinic application prospects.

The invention provides an application of a compound 69-0 and/or tautomers and/or pharmaceutically acceptable salts thereof to prevention, alleviation and/or treatment of COVID19/SARS-CoV-2. Products containing the compounds can effectively inhibit replication or reproduction of SARS-CoV-2 or homologous variant viruses of SARS-CoV-2 and the cytopathic effect produced by SARS-CoV-2. The compound 69-0 has good curative effect, good safety and low toxic and side effects. The invention also provides combined therapeutic pharmaceutical composition. The combined therapeutic pharmaceutical compositioncan effectively inhibit proliferation of SARS-CoV-2 in cells, and has good curative effect, low safety and low toxic and side effects, can reduce drug resistance of viruses and has obvious synergistic effect.

The invention provides a preparation method of a binary inactivated vaccine against a Japanese encephalitis virus and a porcine parvovirus, which comprises the following steps of: (1) inoculating the animal cells to a microcarrier for performing the absorption culture and multiplication culture processes of the cells in a tidal biological reactor; (2) inoculating the Japanese encephalitis virus and the porcine parvovirus on the cells being subjected to multiplication culture; (3) when the inoculated animal cell CPE (Cytopathic Effect) reaches above 75%, harvesting a viral solution; and (4) freezing and thawing the harvested viral solution twice at minus 20 DEG C, inactivating after the concentration, and adding an emulsifier for emulsification to obtain the binary inactivated vaccine against Japanese encephalitis virus and porcine parvovirus. The binary inactivated vaccine provided by the invention has the advantages of good immunogenicity, stable growth and high titer of the preserved seed virus; and the tidal biological reactor is adopted to respectively culture and harvest two viruses with high titers, and the two viruses are directly emulsified and prepared after the concentration and the inactivation, so that the binary inactivated vaccine can be continuously produced in a large scale.

The invention discloses a preparation method and application of enterovirus (EV) 71 type full-length infectious clone with tags. The preparation method comprises the following steps of: (1) extracting total RNA; (2) performing reverse transcription-polymerase chain reaction (RT-PCR) amplification on the EV71; (3) constructing EV71 subclone; (4) constructing recombinant clone containing EV71 genes; and (5) constructing the enterovirus 71 type full-length infectious clone with eGFP and DsRed. Tests, such as cytopathic effect, fluorescence detection, gene detection, virus plaque, medicine inhibition and the like, prove that the EV71 full-length infectious clone with the tags is obtained successfully. The invention has high application value in the aspects of research and development of animal models, virus replication and pathogenesis, medicine screening and medicine action mechanism, vaccine and diagnostic reagents and the like.

The invention provides a cell culture method for duck flavivirus. The method comprises the following steps: inoculating to attach an anchorage-dependent kidney cell BHK21 of a baby hamster to the surface of a cell culture container to form a monolayer; inoculating the monolayer with virus; culturing the monolayer inoculated with the virus in an incubator with 5% CO2 at 37 DEG C; and continuously passing by taking a cell culture product as an inoculum for the next passage. According to the cell culture method provided by the invention, after the virus is continuously passed for 5-7 generations on the monolayer, stable multiplication can be obtained; and after the cell is inoculated for 3-4 days, an obvious cytopathic effect can be formed. The method provides a convenient and visual virus operating platform for studying the biological characteristics of the virus and the molecular basis of virus pathopoiesia and for the vaccine research developed for effectively preventing and controlling the disease.

The invention discloses a TK, ge and gT gene deletion strain of a porcine pseudorabies virus (PRV) variant ZJ01 and application of the TK/ ge/gT gene deletion strain. The strain is the TK/ ge/gT gene deletion strain ZJ01delta TK/gE/gI (ZJ01R in short) of the PRV ZJ01 strain. According to the in-vitro passage test results, gE/gI gene deletion strains ZJ01delta gE/gI of a recombinant virus ZJ01R and the PRV ZJ01 strain are consistent with cytopathic effect generated by a parental virus ZJ01, and virus titer is similar with the ZJ01 strain; a target gene deletion region of the recombinant virus ZJ01R continuously and stably exists without resilience mutation; according to the pig immunity and attacking protection test, after the recombinant virus ZJ01R is immune, a pig is free of clinical symptoms and pathological changes; after being immunized by 21 days, the pig can generate a PRV neutralizing antibody with relatively high level, and can resist challenging infection of a fatal PRV variant strain ZJ01.

The invention provides a construction method of a SOD (superoxide dismutase) gene modified mesenchymal stem cell. The construction method comprises the following steps: (A) connecting an SOD3 gene to an adenovirus vector to obtain a recombinant adenovirus vector; B) co-transfecting the recombinant adenovirus vector and a packaging plasmid to a 293 cell line by utilizing a transfecting agent, culturing until the 293 cell line completely generates a cytopathic effect, collecting an adenovirus strain, storing the strain and confirming that the SOD3 gene is connected to the adenovirus vector; C) amplifying the adenovirus strain, infecting the 293 cell line, collecting the adenovirus supernatant, and determining the titer after ultrafiltration, purification and concentration; and D) infecting the mesenchymal stem cell in different conditions respectively by the adenovirus, and determining the best infection condition to obtain the SOD gene modified mesenchymal stem cell. The EC-SOD (extracellular superoxide dismutase) gene over-expressed SOD gene modified mesenchymal stem cell can be prepared by the method, and can be applied to treatment of mouse with radiation so as to prolong the survival period of the radiated mouse.

The invention discloses a method for building a 7-day-old mouse model infected with enterovirus (EV) 71, comprising the following steps: taking 7-day-old suckling mice and injecting liquid virus into the brains of the mice, thus obtaining the mouse model infected with EV 71. The injection volume of the liquid virus is 20mu L and the liquid virus is prepared by the following steps: inoculating the EV71 seed into MA104 cells in terms of inoculation amount of 200mu L/100mL cell culture bottle, culturing the MA104 cells in 5% of CO2 at 37 DEG C and observing the cytopathic effect every day; on the second day after inoculating EV71, observing that about 70% of the MA104 cells are agglutinated and crimpled, freeze-thawing the cells in the whole bottle and then centrifuging the cells and taking the supernatant, thus obtaining the liquid EV71. The method successfully builds the stable 7-day-old mouse model infected with EV71, provides foundations for the scientific research personnel to research the EV71 etiology and pathogenic mechanism and for screening efficient antivirus drugs and developing vaccines and has broad application prospect.

The invention discloses the application of an andrographolide C15 substitution derivative in manufacturing an anti-hepatitis drug, which belongs to the technical field of pharmaceutical chemistry. According to the invention, MDBK (mardin-darby bovine kidney cells) and BVDV (bovine viral diarrhea virus) are utilized to study the inhibitory action of the compound on cytopathic effect caused by the virus through an external screening model. Through the screening of a large number of andrographolide derivatives, the compound with a structure of general formula 1 is found to significantly inhibit MDBK cytopathic effect caused by the BVDV; the andrographolide C15 substitution derivative has high efficiency and low toxicity, thereby being used for manufacturing medicaments for treating and preventing hepatitis and having good prospect in development and application. The general formula 1 is shown in the description.

The invention belongs to the field of antiviral drugs and provides application of two nitrogen-containing heterocyclic ester compounds to preparation of drugs with resistance to enterovirus 71 (EV71). The nitrogen-containing heterocyclic ester compounds are compounds expressed as formulas WY209 and WY210. Through research experiment of inhibition of novel nitrogen-containing heterocyclic ester compounds on activity of EV71, WY209 and WY210 can inhibit cytopathic effect (CPE) generated by EV71 in host cells RD; the cell survival rate is increased; the progeny virus yield is reduced; the apoptosis of the host cells caused by infection of EV71 can be inhibited; the application shows that the nitrogen-containing heterocyclic ester compounds have potentials for preparing the drugs with resistance to the EV71 virus; the synthesis process of the compounds is simple, economical and rapid; the large-scale production of the compounds is easy to implement; the compounds have clinical application prospects.

The present invention is drawn to generating attenuated and less cytopathic forms of New World alphaviruses that can be used in immunogenic compositions as vaccines against both Old and New World alphaviruses. In this regard, the present invention discloses that the N-terminal, ˜35-aa-long peptide of VEEV, EEEV and, most likely, of WEEV capsid proteins plays the most critical role in the downregulation of cellular transcription and development of cytopathic effect. The identified, VEEV-specific peptide, C_VEE30-68, includes two domains with distinguished functions. The integrity of both domains determines not only the intracellular distribution of C_VEE, but is also essential for direct capsid function in the inhibition of transcription. The replacement of the N-terminal fragment of C_VEE by its SINV-specific counterpart in VEEV TC-83 genome does not affect virus replication in vitro, but makes it less cytopathic and more attenuated in vivo.

The invention relates to a gramine analogue with a heterocyclic structure and application of the gramine analogue with the heterocyclic structure to preparation of anti-CVB3 virus medicines.A chemical formula of the gramine analogue with the heterocyclic structure is as shown in the specification.According to anti-CVB3 activity research experiments of the gramine analogue with the heterocyclic structure, anti-CVB3 pharmacological researches and experiments of CVB3 replication inhibition effect of an analogue G19, the gramine analogue with the heterocyclic structure inhibits CPE (cytopathic effect) of CVB3 on a host cell Hep-2, increases cell survival rate, inhibits progeny virus production and viral nucleic acid generation, mainly inhibits an early replication stage of CVB3 viruses in the host cell and can be applied to preparation of the anti-CVB3 virus medicines.

An object of the present invention is to find a new oncolytic virus and to utilize it for treatment of cancers. The present invention relates to a pharmaceutical composition for treatment of cancers containing Sindbis virus as an active ingredient, a method for treatment of cancers, which comprises administering a therapeutically effective amount of Sindbis virus to a mammal having cancers, a method for identifying cancers, which comprises introducing Sindbis virus into an animal, thereby detecting a Sindbis virus protein or Sindbis virus RNA in vital tissues of the above animal, a method of identifying cancers, which comprises introducing into an animal Sindbis virus having a reporter gene incorporated therein, thereby, detecting the product of said reporter gene in vital tissues of said animal, and a method of identifying cancers, which comprises detecting a increase in antibody titer to the Sindbis virus protein or the product of the reporter gene in the aforementioned animals. It is confirmed that Sindbis virus has cytopathic effects upon various cancer cell lines in vitro, and that reduction in tumor size or disappearance of tumor takes place in vivo. Therefore, Sindbis virus is effective for treatment of cancers. In addition, Sindbis virus shows selective proliferation in a tumor, and is effective for identification or diagnosis for cancers within the living body.

The invention belongs to the technical field of antiviral drugs, and relates to use of novel pyridoheterocyclic ester compounds in manufacture of anti coxsackievirus B3 (CVB3) drugs. The pyridoheterocyclic ester compounds are as shown in formulas WY89, WY94, W95, WY96 and WY97. Through anti CVB3 activity study experiments, the WY89, the WY94, the W95, the WY96 and the WY97 can inhibit cytopathic effect (CPE) produced by CVB3 in host cell Hep-2, can enhance cell viability, reduces progeny virus production, and inhibits of host cell apoptosis caused by CVB3 infection. The anti CVB3 activity study experiments show that the novel pyridoheterocyclic ester compounds have potential in preparation of anti CVB3 infection specific therapeutic drugs, and the novel pyridoheterocyclic ester compounds are simple, rapid and economic in synthesis process and easy in mass production, and have prospects of clinical application.

The invention discloses a preparation method and application of EV71 full length infectious clone with a luciferase tag, belonging to the field of biotechnology. The preparation method comprises the steps of: (1) extracting total RNA; (2) amplifying EV71 through RT-PCR; (3) constructing an EV71 subclone; (4) constructing a recombinant clone containing an EV71 gene; and (5) constructing an EV 71 full length infectious clone with a Rluc tag. Experiments of cytopathic effect, Rluc activity detection, virus plaque, drug inhibition and the like show that the EV71 full length infectious clone with a tag is successfully obtained. The invention has wide application values in the aspects of animal models, virus replication and pathogenesis, drug screening and mechanism of drug action, development of vaccines and diagnostic reagents and the like.

The invention discloses a recombinant virus of a chimeric IBV H120 S1 gene ectodomain suitable for cell culture and a construction method and application thereof. The recombinant virus is constructed by the following steps of: replacing the S1 gene ectodomain segment of an IBV Beaudette strain with an S1 gene ectodomain segment of an IBV H120 strain to construct the recombinant virus; after successful virus rescue, passing from the Vero cell; collecting toxicity when the cell cytopathic effect area is over 90%; and repeatedly freezing and thawing for three times, and continuing passage to obtain the recombinant virus of a chimeric IBV H120 S1 gene ectodomain suitable for cell culture. The recombinant virus disclosed by the invention is used as a vaccine strain to immunize SPF chicken; after counteracting toxicity, the chicken flocks of an immunity group and a control group are continuously observed to discover that the recombinant virus disclosed by the invention can serve as a vaccine strain to provide good immunity protection to the inoculated chicken; and the recombinant virus is safe to various kinds of chicken and avoids side reaction.