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Objective To observe the effects of intranasal inoculation with recombinant Mycobacteri-um smegmatis ( rMS) on the airway inflammation of non-eosinophilic asthmatic mouse and further investigate the relative mechanism. Methods DO11. 10 T-cell receptor transgenic mice were divided randomly into three groups. Non-eosinophilic asthmatic model was established via OVA challenge, rMS were administrated into mice before challenge. Anti-IL-17A autoantibody in sera, IL-6 and IL-17A in BALF were measured by ELISA, the proportion of neutrophil in BALF was measured by FCM, MPO activity in lung tissue was detected by colorimetry, and the mRNA expression of CXCL2 and CXCL5 was measured by real-time PCR. Results Compared to control group, the number of neutrophils, IL-6 and IL-17A levels in BALF from asthmatic mice was significantly increased, meanwhile MPO activity and the expression of CXCL2, CXCL5 in lung tissue were both significantly upregulated. The results showed that high titer of autoantibody of IL-17A in sera of mice vaccined with recombinant Mycobacterium smegmatis was detected. Compared to asthma group, neutrophils and IL-6, IL-17A in BALF were significantly decreased, meanwhile MPO activity and CXCL2, CXL5 mRNA expression were significantly down-regulated. Conclusion Recombinant Mycobacterium smegmatis exhibits anti-inflammatory activity in murine neutrophilic asthma model and it may have protective effects on asthma.
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Objectives To analyze the clinical characteristics and signiflcance of Mycoplasma pneumoniae pneumonia (MPP) combined with secondary thrombocytosis in infants and young children. Methods Clinical features, laboratory and imaging data of the infants and young children with MPP were collected, and compared between the two groups of children with and without secondary thrombocytosis. Results Sixty-seven (67) infants and young children with secondary thrombocytosis (28 males and 39 females, with onset at 2 years and 2 months after birth in average) and 269 infants and young children with normal platelet counts (138 males and 131 females, at 2 years and 3 months after birth in average) were included. Signiflcant longer durations of hospitalization(P =0.018) and fever(P =0.000), higher temperature peak(P =0.000), as well as higher morbidity of refractory MPP(P =0.001) and more complications(P =0.000)were observed in the group of MPP with secondary thrombocytosis. Moreover, the laboratory data of white blood cell count(P =0.000), C-reactive protein(P =0.000),procalcitonin (P =0.000), erythrocyte sedimentation rate(P =0.000)and higher morbidity of multiple pathogen infection(P =0.033) were observed between the two groups.Conclusions More severe clinical manifestations and higher complication morbidity could be observed in the group of MPP combined with secondary thrombocytosis, implicating the value of platelet count in clinical assessment of MPP in infants and young children at early stage.
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Objective To identify the dynamic influence of antibiotics use on T helper cell(Th) 1/Th2 cytokine balance in neonatal rats in their early life.Methods Twenty-four newborn SD rats were randomly divided into 3 groups including 2dCef group,7dCef group and a control group (n =8).SD rats at 2 or 7 days of age were injected Cefotaxime sodium intraperitoneally for 3 consecutive days.The control group was injected the same amount of phosphate buffer saline.Serum and bronchoaveolar lavage fluid (BALF) samples were collected on 35 days old and diffe-rential cell count in BALF was done.Enzyme linked immunosorbent was used to detect the concentration of interleukin(IL)-12,interferon-γ(IFN-γ),IL-4 and IL-5 in serum and BALF,and real-time quantitative PCR was used to detect mRNA expression of those cytokines in lung tissues.Results The number of total cells,neutrophils,and lymphocytes in Cefotaxime treated on 2nd day after birth were significantly higher than those in the controls and 7th day group (F =93.78,15.84,7.36,all P ＜0.05).For Thl cytokines detection:the IL-12 secretion in BALF and serum and INF -γsecretion in BALF of the 2 antibiotic groups were lower than those in control group (F =26.53,12.95,47.18,all P ＜ 0.05),the decrease in 2dCef group were more obviously than 7dCef group [(127.72 ± 12.35) ng/L vs (198.15 ± 18.93) ng/L,(105.91±13.61) ng/Lvs (133.63 ±13.82) ng/L,(147.23 ±24.10) ng/Lvs (178.52±12.46) ng/L,all P ＜0.05].The mRNA expression of IL-12 in antibiotic groups were lower than those in control group (F =33.60,P ＜0.05),and the decrease in 2dCef group was more obvious than that in 7 dCef group (0.073 ± 0.023 vs 0.093 ± 0.028,P ＜0.05).For the Th2 cytokines:the IL-4 secretion concentration and mRNA expression in the 2 antibiotic groups were higher than those in the control group (BALF,serum and lung) (F =64.26,13.79,11.54,all P ＜ 0.05),the increase in 2dCef group was more obvious than that in 7dCef group[(103.65 ± 7.68) ng/L vs (82.04 ± 8.77) ng/L,(87.81 ± 4.72) ng/L vs (69.69 ± 7.79) ng/L,(0.067 ± 0.036) ng/L vs (0.051 ± 0.055),all P ＜ 0.05];however,no significant differences of IL-5 either mRNA or cytokine secretion being detected among these 3 groups (F =0.50,0.75,0.08,all P ＞ 0.05).Conclusions Antibiotic use during early time of neonate rats alters postnatal immune cells distribution in the lung and promotes the shift in cytokine balance towards a Th2 profile,and the earlier the antibiotic use,the more serious the impact.
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Objective To evaluate the effects of recombinant fusion protein interleukin-17F/His (IL-17F/His) on expression of immune-related inflammatory factors in murine models with Streptococcus pneumoniae (SP) infection.Methods BALB/c mice were randomly divided into SP infection group,fusion protein intervention group and the normal control group.Before intranasal infection with SP,the mice were treated with PBS or IL-17F/His respectively.The number of bacteria and leukocytes in bronchoalveolar lavage fluid (BALF) was counted,and the mRNA levels of β-defensin-2 (Defb2),macrophage inflammatory protein (MIP)-lα and MIP-2β in lung tissue were detected by real-time fluorescent quantitative PCR,the concentrations of MIP-1 α,MIP-2β,IFN-γ and IL-4 in BALF,supematants of spleen cell and mediastinal lymph node cell were assayed by enzyme linked immunosorbent assay.The expressions of IL-17F and Defb2 in lung tissues were observed with immunocytochemistry.Results 1.Compared with the SP infection group,in the fusion protein intervention group,the total number of WBC (209.00 ± 18.23) was higher,the number of neutrophil,macrophage and lymphocyte[(8.67 ±2.16) × 106/L,(193.50 ± 16.50) × 106/L,(6.83 ± 1.17) × 106/L] were higher in BALF(U/F =38.097,28.854,32.942,27.810,all P ＜ 0.05),while the number of bacteria [2.75 (1.15-3.09) × 103/L] was significantly lower(P =0.02).2.Compared with SP infection group,the expression levels of Defb2 mRNA and MIP-1α mRNA(31.64 ±4.97,5.81 ± 1.09) in lung tissues were higher in the fusion protein intervention group,while the expression level of MIP-2β mRNA (6.69 ± 2.05) was lower (t =4.889,2.306,3.536,all P ＜ 0.05).3.Compared with the SP infection group,the concentration of MIP-2β [(64.15 ± 13.41) ng/L] was significantly decreased and concentration of IL-4 [(92.28 ± 4.52) ng/L] was significantly increased in BALF in the fusion protein intervention group(H/F =159,289,40.767,all P ＜0.01).4.Compared with the SP infection group,the concentrations of MIP-2β and IL-4 [(255.02 ± 13.95) ng/L,(107.02 ± 7.53) ng/L] were both significantly increased in spleen cell culture supematants in the fusion protein intervention group,the concentrations of MIP-1 α and IFN-γ [(413.61 ± 24.23) ng/L,(98.88 ± 5.84) ng/L] were both significantly decreased (all P ＜ 0.05).5.Immunocytochemistry analysis revealed that the expressions of IL-17F and Defb2 were both higher in the fusion protein intervention group than those in the SP infection group; there was no expression in the normal control group.Conclusions Intranasal inoculation of recombinant fusion protein IL-17F/His can promote pulmonary neutrophil and macrophage recruitment and increase bacterium clearance,and enhance the host defense against SP infection through increa-sing the expression of defensins,regulating the levels of MIP,IL-4 and IFN-γ.
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ObjectiveTo evaluate the effects of intranasal administration of recombinant interleukin-17A(rIL-17A) on the expressions of β-Defensin-2(Defb2) and macrophage inflammatory protein(MIP) in pneumococcal pneumonia murine models.MethodsTwenty-four BALB/c mice were divided randomly into normal control,pneumococcal pneumonia,and rIL-17A intervention groups ( n =8 ).Before intranasal (i.n) infection with Streptococcus pneumoniae,the mouse was treated with PBS or rIL-17Ai.n respectively.The mRNA levels of Defb2,MIP-1α and MIP-2β expression in lung tissue were detected by real-time quantity PCR.The numbers of bacteria and leukocytes in bronchoalveolar lavage fluid (BALF) were counted as well.And the concentrations of MIP-1α,MIP-2β,IFN-γ and IL-4 in BALF and in supematants of spleen cells and mediastinal lymph node cells were assayed by ELISA.Changes in lung tissue histopathology were observed with HE staining through light microscope.ResultsNeutrophil and macrophage numbers are higher in BALF of rIL-17A group,while the numbers of bacteria were lower,when compared with those in pneumonia group( P＜0.01 ).The expression of Defb2 and MIP-1α mRNA were up-regulated in lung after rIL17A treatment(P＜0.01 ).When compared with rIL-17A non-treated mice,rIL-17A treated mice secretedhigher levels of MIP-1α in lymph node cell culture supernatants( P＜0.01 ),higher levels of MIP-2β were observed in spleen cell and lymph node cell culture supernatants( P＜0.01 ),higher levels of IFN-T were detected in BALF( P ＜ 0.01 ) and culture supernatants of spleen cell ( P ＜ 0.01 ) and lymph node cell ( P ＜0.05),and higher levels of IL-4 were detected in BALF and spleen cell culture supernatant(P＜0.01 ).Comparative analysis have not detect a significant irflammatory cell increases in rIL-17A treated mice lung tissue; however the histopathological lesions were decreased.ConclusionIntranasal inoculation of rIL-17A can promote pulmonary neutrophil and macrophage recruitment and bacterium clearance,Intranasal inoculation of riL-17enhances the host defense against Streptococcus pneumoniae infection partly through increasing the expression levels of defensins,MIP,IFN-T and IL-4 etc.
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The application of standardized patien(tSP)has not been widely used in China. This article summarized some experiences of using SP in pediatric clinical teaching such as choice and training SP,the writing of scenario,and the score criterion in OSCE. We concluded that the use of SP had irreplaceable advantages.