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Objective@#To explore the therapeutic effects of dendritic cells (DC) modified by the dust-mite-allergen(Der p1) gene on mouse model of allergic rhinitis (AR).@*Methods@#DC modified by the Der p1 gene (Der p1-DC) were prepared.Using random number table, 24 Balb\c mice were divided into four groups: immature DC (imDC)/AR group, dexamethasone/AR group, Der p1-DC/AR group and control group, with 6 mice in each group.AR mouse model was built with Der p1 and the mouse model of AR was established.The AR mice were respectively given by abdominal injection of Der p1-DC, imDC and dexamethasone.Normal control mice were treated with physiologic saline.ELISA method was used for determining the content of IgE, IgG1and histamine in blood.The relative expression of mRNA of IL-4 and IL-13 on nasal mucosa with protein was analyzed by RT-PCR and Westen blot methods.All the data were statistically analyzed by SPSS 19.0 statistical software, and the variance analysis was used in multiple groups of average samples.@*Results@#The contents of IgE, IgG1 and histamine in the mice of Der p1-DC/AR group were lower than those in imDC/AR group ((0.560±0.110) OD 450 nm vs (1.150±0.280) OD 450 nm, (0.690±0.054) OD 450 nm vs (0.920±0.125) OD 450 nm, (4 145±670) pg/ml vs (7 685±669) pg/ml, t value was 4.80, 4.14, 9.16, respectively, all P<0.05), and the expression of IL-4 and IL-13 on nasal mucosa in Der p1-DC/AR group was remarkedly lower than those in imDC/AR group (0.41±0.25 vs 1.59±1.02, 0.26±0.01 vs 1.10±0.09, t value was 2.75, 22.72, respectively, all P<0.05). There was no statistically significant difference between the mice treated with Der p1-DC and dexamethasone group.@*Conclusions@#The results showed that Der p1-DC could reduce inflammation in AR mice and decrease the expression of IL-4 and IL-13. It suggested that Der p1-DC can be used in the immunotherapy of AR mouse.
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Aim To observe the expression of epider-mal growth factor receptor ( EGFR) in cerebral tissues around hematomas after intracerebral hemorrhage, and explore the effects of EGFR on activation of astrocytes derived from rats and the involved mechanisms. Meth-ods The specimens of cerebral tissues around hemo-tomas after intracerebral hemorrhage undergoing hemo-tomas removal operation were collected and then divid-ed into 4 groups according to the time of intracerebral hemorrhage: 10 d groups. Each group included 20 cases. At the same time, 20 dropped brain tissues distant to hemorrhage in the operative process were collected as control group. Immunohistostaining and Western blot were used to measure the expression of EGFR. After isolation and culturing, the astrocytes of rat cortex were treated with culture solution ( control group) , CNTF that was used to activate astrocytes, scramble siRNA + CNTF and EGFR siRNA +CNTF for 24h, respectively. The ex-pression of glial fibrillary acidic protein ( GFAP) mR-NA was detected through fluorescence real-time quanti-tative PCR. In addition, the protein levels of GFAP, signal transducers and activators of transcription 3 ( STAT3 ) and phosphorylated STAT3 ( p-STAT3 ) were examined using Western blot. Results With the ex-tension of intracerebral hemorrhage time, positive sig-nal index and protein expression levels of EGFR gradu-ally elevated, reached the peak on 6 ~10d, and then decreased after 10 d. There was statistical difference ( P0. 05 ) . Conclusions EGFR expression is upregulated in the cerebral tissues around hemotomas after intracerebral hemorrhage. Gene silence of EGFR contributes to suppressing the activation of astrocytes derived from rats, which may be involved in the block-ade of STAT3 phosphorylation.
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Dendritic cells (DCs) is known as the most potential and professional antigen presenting cells (APC), it mainly involves in the cellular immunity and T cell dependent humoral immunity, which plays a key role in the immune response and is one of the most hot areas in immunology in recent years. DCs plays a key role in allergic rhinitis (AR) and is one of the most important mechanism of AR treating by sublingual immunotherapy (SLIT). This article reviewed the mechanism of the role of DCs in AR and AR treating by SLIT.
Subject(s)
Animals , Humans , Dendritic Cells , Allergy and Immunology , Desensitization, Immunologic , Rhinitis, Allergic , Therapeutics , Sublingual ImmunotherapyABSTRACT
A case of vagus nerve invasion with disseminated herpes zoster was reported. Clinical manifestation of disseminated herpes zoster and vagus nerve injury. relevant imaging examination and laboratory examination can help to establish a preliminary diagnosis. Anti-virus, anti-infection and symptomatic treatment had been performed and showed good clinical efficacy.
Subject(s)
Aged , Humans , Male , Herpes Zoster , Pathology , Vagus Nerve , PathologyABSTRACT
Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.
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ObjectiveTo develop a transgenic mouse model for N.gonorrhoeae researches.Methods Human carcinoembryonic antigen-related cellular adhesion molecules 1 (hCEACAM1) eukaryotic expression vector,pCDPGICAM1,was used to generate transgenic mice by microinjection.The funder mice were screened by PCR,sequence analysis,Western blot and fluorescence-activated cell sorting analysis,respectively.The transgenic mice expressing hCEACAM1 were inoculated with N.gonorrhoeae intravaginally.Adhesion and infection of gonococci to mice were analyzed by bacteria culture and microscopy.Results Four (lines 50,53,54,and 59) of the 22 F0 generation transgenic mice were found to carry the transgene.The hCEACAM1 protein was expressed on the cell membrane of various tissues in the line 53 transgenic mouse.Compared with normal mice,N.gonorrhoeae can successfully infect and cause inflammation in the transgenic mice.Conclusion The hCEACAM1 transgenic mouse can be used as an animal model for gonococcal infections.
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By combining the teaching environment and taking the multimedia teaching system as the platform,we adopted the nimble utilization teaching method in physiology teaching process which fully aroused the students'enthusiasm in studies,and played the certain impetus role to students' quality enhancement.
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AIM: To prepare gfp-bcl-X L-contained recombinant adenovirus(rAd-gfp-bcl-X L).METHODS: Bcl-X L gene was amplified from pEGFP-C 3-bcl-X L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X L. Then pAdTrack-CMV-bcl-X L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X L gene. The titer of the purified rAd-gfp-bcl-X L was 6 5?10 12 PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X L. This affords a good gene transfer vector for the gene therapy in human's diseases.