Objective:To prepare recombinant pCDNA3.1+/Ag85A DNA vaccine encoding tuberculosis Ag85A gene mediated with LipofectamineTM 2000 and to study the effect on T cell subpopulation via oral vaccination.Methods:Recombinant plasmid containing Ag85A using liposome as a vector was constructed and administered to C57BL/6 mice via oral route.Determination of the contents of IFN-r,IL-4 in serum of C57BL/6 mice was performed by double antibody sandwich ELISA.Furthermore,the ability of splenocytes to secret IFN-? and IL-4 was tested by ELISPOT method.When the mice were vaccinated with recombinant eukaryotic expressing vector 5 weeks later,titers of serum antibody against Ag85A were detected by ELISA.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was determined by flow cytometry.Results:Lymphocytes obtained from the spleen of liposomal pcDNA3.1+/Ag85A vaccine-immunized mice exhibited lower IFN-? production and higher IL-4 production than those of pcDNA3.1+ vector immunized mice.The number of spleen MNC secreting IFN-? stimulated by Ag85A protein in vitro was significantly lower than that of plasmid vector group.Liposomal pcDNA3.1+/Ag85A vaccine immunized mice elicited higher Ag85A-specific antibodcy titres.The percentage of the CD4+ and CD8+ T cell subsets in the splenocytes was decreased.The subset both were shown the profile of Th2 responses.Conclusion:The oral recombinant plasmid pCDNA3.1+/Ag85A mediated with LipofectamineTM 2000 It shows down-regulation effect on the subsets of CD4+ T cells and CD8+ T cells.The regimen has good immunogenecity and could induce Th2 type humoral response in C57BL/6 mice.The immunization suppresses secretion of IFN-?.It can greatly enhance the titres of Ag85A-specific antibodies.LipofectamineTM 2000 can act as an adjuvant through comparation with negative control group.

OBJECTIVE: To investigate the expression and role of Interleukin-33 (IL-33) and ST2 in the nasal polyps of human Eosinophilic and non-Eosinophilic chronic rhinosinusitis with nasal polyps (ECRS and non-ECRS). METHOD: IL-33 and ST2 protein expression in nasal polyps of ECRS and non-ECRS as well as in seemingly normal mucosa of the inferior turbinate tissue was investigated by immunohistochemical staining and messenger RNA (mRNA) expression of IL-33 and ST2 was assessed by realtime polymerase chain reaction (PCR) in 27 subjects with ECRS, 33 subjects with non-ECRS, and 11 control subjects. RESULT: (1) The ST2 was found both in nasal polyps of ECRS and non-ECRS,especially in ECRS, yet hardly found in the normal mucosa of the inferior turbinate tissue; (2) The expression of ST2 mRNA in nasal polyps of ECRS was higher than that in non-ECRS and normal inferior turbinate tissue, and the difference was both prominent in statistics (P<0.01); (3) The expression patterns of IL-33 at both mRNA and protein levels were not significantly different among the three groups (P>0.05). CONCLUSION The IL-33 and its receptor ST2 were both expressed in human nasal polyps including ECRS and non-ECRS, meanwhile the expression patterns of ST2 at both mRNA and protein levels were significantly higher in nasal polyps of ECRS. The current study suggests that IL-33 and its receptor ST2 may play important roles in the pathogenesis of chronic rhinosinusitis with nasal polyps, especially in ECRS through the increased expression of ST2 in Eosinophils as a hypothesis.
Chronic Disease ; Eosinophils ; immunology ; Humans ; Interleukin-1 Receptor-Like 1 Protein ; Interleukin-33 ; metabolism ; Nasal Mucosa ; metabolism ; Nasal Polyps ; immunology ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Receptors, Cell Surface ; metabolism ; Rhinitis ; immunology ; Sinusitis ; immunology ; Turbinates ; metabolism

Objective:To investigate the effect of perforin-mediated cytotoxicity in primary influenza virus infection.Methods:Perforin-deficient and wild-type C57BL/6 mice were infected intranasally with influenza virus A/PR/8/34. Pulmonary viral growth was determined at various days after infection by pfu experiment. Perforin-mediated apoptotic degeneration was observed by Immunohistochemical staining. LDH-release method was used for detection of specific CTL and NK cell activity from spleen cells.Results:Mice deficient in the perforin gene showed an increased virus growth and prolonged virus shedding. The appearance of apoptotic degeneration in virally infected lung cells was delayed in perforin-deficient mice. The cytolytic activities of natural killer cells and virus-specific cytotoxic T lymphocytes were significantly lower than that of wild-type mice.Conclusion:Perforin plays a critical role in the host defense system against primary influenza virus infection.

Objective:To prepare a colloid gold kit for the qualitative detection of the dopes met-amphetamine and morphine by one-step chromatography immunoassay in human samples based on the principle of the highly specific immunochemical reactions between the antigens and their antibodies which were used for the analysis of specific compounds in human urine samples at ideal cut-off concentrations.Methods:The monoclonal antibodies(McAb) of either mouse anti-morphine or mouse met-amphetamine were prepared by hybridomas and the limiting dilution respectively. The titers of McAb were tested by ELISA.The kit was assembled and the specificity,sensitivity,and stability were investigated.In addition,these characters were compared with those of issued Pan Probe single test strip.Results:①The titer of the first McAb against morphine was 1∶3.2?103, as for another, 1∶1.6?103. ②The colloid gold kit was found with following cut-off for the dope concentrations,morphine at 300 ng/ml and met-amphetamine at 1 000 ng/ml. ③The colloid gold kit showed fine specificity without cross-reaction with routine medicines for cold treatment or with the solf-drinks of cokocola or with tobacco for the individuals who did not take the dopes the 7 days.④The colloid gold kit was even more sensitive than the issued strip,because urine sample of the junkies were still positive 4-5 days post-intake.Conclusion:The colloid gold kit for indentifying the drug-users is a rapid,specific, sensitive immunoassay suitable for the simultaneous, qualitative detection of drug abuse.

Education reform and innovative education are the strong requirements of the times and social development.We developed immunological paper free examination system based on the optimized question bank,characteristics of immunological examination and information technology.This system can randomly develop electronic examination paper and can integrate the processes of exam registration,examination paper development,online examination,paper inspection,scores generation and printing into one system,which can save human resources,enhance the accuracy and fairness of the examination,making it conform to international standards.

Objective To evaluate the therapeutic effects and experience of the tracheal stent implantation for the management of severe tracheal stenosis.Materials Thirteen patients with severe tracheal stenosis of various causes underwent high kilovoltage radiography and computed tomography for evaluating the site,form and extent of the stenosis including 10 at the trachea,1 at the right main bronchus and 2 at left main bronchus.The C2 catheter assisted with ultra-slipping guide wire was inserted into the trachea under fluoroscopy and then a replaced high shoring guide wire was pushed through the stenotic segment and retained the stent. Results All stents were implanted successfully with successful rate 100% together with dyspneic improvements.The mean survival time was 6.2 months for patients with malignant neoplasm.One patient with benign tracheal stenosis has been followed-up for 5 years without restenosis. Conclusions The tracheal stent implantation is an effective means for severe tracheal stenosis.

Objective:To study the activity of APC in early phase infection by detection of the differentiation of Th0 cells from TG mice Methods:Detection of IFN ? and IL 4 by cytokine ELISA Identification of the phenotype of T cells from culture wells by FACS Results:Infected APC induced the T cells of TG mice to differentiate into Th1 cells, uninfected APC induced it into Th2 cells IL 4 and IL 12 influenced the effect of APC on the differentiation of T cells Conclusion:Infection and added cytokines influenced the presenting function of APC

Objective To retrospectively analyze the effect of interventional embolization for hepatocelluar carcinoma (HCC) associated with arteriovenous shunting (AVS), and to discuss the factors influencing the therapeutic results. Methods The clinical data of 62 cases with HCC associated with AVS , who were treated with interventional chemoembolization , were retrospectively analyzed. Based on the type and extent of AVS identified by angiographic manifestations, appropriate obstruction of the shunt and Lipiodol chemoembolization of HCC were conducted. The curative effect of the shunt embolization was assessed by DSA at one or two months after the treatment. The relevant factors influencing the prognosis of embolization were analyzed by using univariate and multivariate Cox regression analysis methods. Results Of the 62 patients, arterioportal shunting (APS) was detected in 44, hepatic arterio-venous shunting (HAVS) in 11, APS together with HAVS in 4, and hepatic artery-pulmonary artery shunting (HAPAS) in 3. Re-examination with DSA was carried out in 53 patients at 1 - 2 months after the treatment , which showed that the shunting disappeared in 18 cases, obvious reduction of the shunt flow was seen in 19 cases, the lesion remained stable in 9 cases and the disease became worse in 7 cases. Univariate analysis indicated that the kind of embolic material and the presence of tumor thrombus could affect the obstructive result of the shunt , while multivariate Cox regression analysis showed that portal tumoral thrombus was an independent risk factor. The embolization effect of polyvinyl alcohol (PVA) particles and Lipiodol-ethanol mixture, used as the embolic agents, was better than that of gelatin sponge particles. Conclusion To ensure a successful interventional chemoembolization for HCC combined with AVS the procedure should be individualized according to the type and extent of the arteriovenous shunting. The type of embolic materials used for embolization can affect the results to a certain degree.

ObjectiveTo evaluate the technical feasibility of animal model of avascular necrosis of femoral head (ANFH)with transcatheter arterial embolization (TAE).MethodsTwenty experimental pigs were randomly divided into experimental group and control group (each n= 10).Experimental group:A 5F Cobra catheter was inserted into left femoral artery,and the feeding arteries of femoral head were superselectively inserted.The feeding arteries were embolismed through transcatheter arterial injecting the segments of silk measuring about 500μm.Control group:The arterial embolization was not performed,and the other treatment was identical to experimental group.The articulation of hip in all pigs underwent plain X-ray examination,CT and MR scanning 2 weeks and 4 weeks after treatment,respectively.Histological examination was made in 4 weeks to evaluate volume of bone trabecula (TBV) and percentage of bone lacuna (PBL) at unit area under microscope.The data were compared between the two groups.Results In experimental group,CT and MRI showed swolling in hip soft tissue and high T1 in hip joint cavity,while no obvious abnormalities were found in plain X-ray film 2 weeks after feeding arteries were embolized.Four weeks after feeding arteries embolization,plain X-ray film,CT and MR showed typical necrosis of femoral head in the experimental group,while no obvious abnormalities were found in control group.The histology examination revealed there were obvious karyopyknosis and anachromasis in the bone cells.The quantity of bone cells decreased obviously or disappeared.PBL increased and TBV decreased significantly compared with those of control group (P＜0.05).ConclusionThe animal model of ANFH in pigs can be induced by TAE.It can preferably mimic the pathological situation of ANFH.

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.