7 results about "Infection experiment" patented technology

The application provides a kind of nasal mucosa in vitro culture method and culture fluid, the composition of the culture fluid includes Wnt / β-catenin signal pathway activator 0.1-20 μg / ml, fibroblast growth factor 10-150 ng / ml, TGF-β inhibitor 0.02-3 μg / ml, ROCK inhibitor 0.1-100 μmol / L, P38 MAPK inhibitor 0.1-1 μmol / L, serum-free additive 1X, N-acetyl cysteine 1-10 mmol / L, nicotinamide 1-20 mmol / L, GlutaMax additive 1X, antibiotic, DMEM / F12 culture medium 1X.The application can carry out in vitro culture to nasal mucosa, can keep at least 2 weeks of activity, and have complete epithelial barrier and immune system, can realize ex vivo nasal mucosa virus infection experiment.

The application provides a method for constructing an immortalized bovine nasopharyngeal tonsil epithelial cell line and application, and belongs to the technical field of cell engineering. The healthy yellow bovine nasopharyngeal tonsil is used as a material, and the primary epithelial cells are separated by enzyme digestion. The 3rd to 4th generation cells are taken, infected with SV40 T lentivirus (MOI=80), and screened by 1.5 μg / mL puromycin to obtain a stable immortalized cell line. The cell line is continuously passaged to the 50th generation and still stably expresses SV40LT protein and epithelial specific markers CD326, CD324 and KRT8, maintains a typical pavement stone-like morphology, normal proliferation and no malignant transformation. Infection experiments show that the cell is highly sensitive to FMDV, supports efficient replication, and the virus titer can reach 10 7.5 TCID 50 / mL. The application overcomes the defects of the existing immortalized bovine nasopharyngeal tonsil epithelial cell model, such as limited source, large batch difference, low sensitivity and poor persistence, and provides a stable and reliable in vitro model for the research of foot-and-mouth disease prevention and control related mechanisms, drug screening and vaccine development.

The application provides an animal immune evaluation method based on a Lutzomyia longipalpis infection model, and comprises the following steps: S1, experimental animal preparation and screening; S2, Lutzomyia longipalpis preparation and quality verification; S3, Lutzomyia longipalpis infection of experimental animals; S4, immune related index detection; S5, animal immune function evaluation; and S6, repeatability verification and data summarization. The application realizes systematic and objective evaluation of the effect of immune candidate drugs.

The application discloses a kind of siRNA and antiviral application of targeting swine epidemic diarrhea virus Nsp9, belong to the field of biotechnology.The application designs a kind of siRNA targeting PEDV Nsp9, the nucleotide sequence of the siRNA sense strand is as shown in SEQ ID NO.3, the nucleotide sequence of the siRNA antisense strand is as shown in SEQ ID NO.4.It is proved by cell transfection experiment and virus infection experiment that the siRNA prepared in the application can significantly reduce the replication infection of PEDV in cells, effectively inhibit the expression of virus, and play an antiviral role;And the siRNA has no cytotoxicity, and has extensive application prospect.

The utility model discloses a centrifugal device for DF1 cell infection experiment, which comprises a support frame and a support disc connected to the top of the support frame, the center of the support disc is rotatably connected with a centrifugal shaft, a support plate is connected with a spring I for supporting the support plate, the top of the centrifugal shaft is connected with a pressing mechanism for pressing a plurality of test tubes, and the spring I is connected with a spring II for supporting the support plate. The pressing mechanism comprises a connecting rod, a rotating cylinder, a movable disc, a sliding block, a second spring, a plurality of L-shaped plates and a plurality of glands, the two ends of each L-shaped plate are connected with the rotating cylinder and the corresponding glands, the sliding block is used for guiding the movable disc, and the second spring is used for driving the movable disc to move in the direction close to the centrifugal shaft. The top of the centrifugal shaft is connected with a locking mechanism used for locking the L-shaped plate. The stability of a plurality of glands can be improved, a plurality of test tubes can be synchronously pressed and fixed, the installation time of the test tubes is shortened, and the working efficiency is improved.

The invention provides an animal immunity evaluation method based on a chlamys farreri infection model. The animal immunity evaluation method comprises the following steps: S1, preparing and screening experimental animals; s2, preparation and quality verification of the chlamys farreri; s3, infecting experimental animals with the chlamys farreri; s4, detecting immune related indexes; s5, animal immune function evaluation; and S6, carrying out repeatability verification and data summarization. According to the invention, systematic and objective evaluation of the effect of the immune candidate drug is realized.

This invention provides the application of norzelamin in the preparation of NDM-1 enzyme inhibitors. Through cefotaxime hydrolysis assays, checkerboard minimum inhibitory concentration (MIC) tests, growth curves, bactericidal curves, hemolysis tests, and *Gnaphalium affine* infection experiments, it was demonstrated that norzelamin can inhibit NDM-1 enzyme activity and exhibits good synergistic antibacterial activity with the carbapenem antibiotic meropenem, enhancing the protective effect of meropenem against *Gnaphalium affine* larval infection models. Therefore, norzelamin, as a potential adjuvant for β-lactam antibiotics, can enhance the antibacterial effect of β-lactam antibiotics by inhibiting NDM-1 enzyme activity, which is of great significance for the clinical development of safe and effective β-lactam antibiotic adjuvants.