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Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine
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Objective@#To investigate the pathogenic characteristics of viral encephalitis in children living in Hebei province.@*Methods@#We randomly collected cerebrospinal fluid specimens from a total of 399 children diagnosed with viral encephalitis in Hebei Children′s Hospital from May to December 2017. Real-time fluorescence quantitative PCR and Sanger sequencing were used to detect viral nucleic acids in cerebrospinal fluid by an automatic laboratory station. Statistical analysis was performed on the experimental data using SPSS 21.0 software and the clinical data were analyzed. Comparison of infection rates of EV encephalitis in different months, using line × column chi-square test. The MRI and EEG positive rates of different viral encephalitis and viral encephalitis patients not infected with the virus were analyzed by Fisher′s exact probability test. The positive rate of infection with different viruses and non-virus agents was analyzed by Fisher′s exact probability test.@*Results@#The result showed that 80 of 399 samples were positive, and the positive rate was 20.05%. It included 22 cases of enterovirus, 4 cases of influenza A virus, 3 cases of mumps virus, 2 cases of herpes simplex virus type 1, 1 case of herpes simplex virus type 2, 4 cases of EB virus, 7 cases of cytomegalovirus, 7 cases of herpes zoster virus, 8 cases of adenovirus, 14 cases of human herpesvirus type 6. Eight cases had combined viral infection. Eight cases had concurrent infections: 3 cases had enterovirus and herpesvirus type 6 concurrent infection, 1 case had enterovirus and Japanese encephalitis virus concurrent infection and 1 case had herpes simplex virus type 2 and adenovirus, 1 case had influenza A virus herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had herpes simplex virus type 1 and herpes zoster virus concurrent infections. Children with EV viral encephalitis in Hebei Province were highly prevalent in May and June (P=0.016). HHV6 virus encephalitis was more susceptible to infection than non-HHV6 virus (P=0.016); The rate of MRI positive findings in patients with different viral encephalitis was not statistically significant (P>0.05). The result of EEG of different viral encephalitis were P>0.05, which was not statistically significant.@*Conclusions@#EV was the most common pathogen of children with viral encephalitis in Hebei province. Encephalitis caused by influenza A virus cannot be ignored in clinical practice.
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Objective To investigate the significance of CD44v6 expression in acute leukemia( AL) and it's relation with the prognosis of AL. Methods Sixty AL patients were treated by enzyme linked immunosorbent assay( ELISA)as initial treatment group. Fourty-seven cases were remission as remission group, and 20 cases with no-remission group. Meanwhile,45 healthy people were served as the control group. The level of CD44v6 was measured by ELISA. Results The serum CD44v6 in initial treatment patients,remission group, no-remission group and control group were( 179. 34 ± 39. 41 )μg/L,( 190. 61 ± 28. 05 )μg/L,( 106. 72 ± 26. 38)μg/L and(98. 31 ± 21. 78)μg/L respectively,and the difference was significant( F =56. 303,P﹤0. 01),and the CD44v6 of initial treatment group and remission group were higher than that of no-remission group and control group(P﹤0. 05). The leukocyte levels was positive related to CD44v6 levels in 60 patients(r=0. 826,P﹤0. 01),and it was also related to disease stage,extramedullary infiltration(( r=0. 485,0. 512;P﹤0. 01). Conclusion The level of CD44v6 is closely related with the occurrence and development of acute leukemia. The assay of CD44v6 in serum of AL patients is helpful in diagnosing and predicting the risk of metastasis and prognosis in AL.
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ObjectiveTo investigate the serum soluble urokinase-type plasminogen activator (suPAR)in patients with multiple myeloma,its relationship with the 2-year survival rate and its clinical significance in predicting the disease prognosis.MethodsEnzyme-linked immunosorbent assay was applied to determine the levels of suPAR of 40 initial diagnosed patients with MM and 30 healthy persons as healthy control group.The patients were divided into three groups:effective (13 cases),remission (19 cases) and invalid (8 cases).ResultsUsually there was suPAR expression [(233.47±83.22) pg/ml] in serum of the healthy adults.There was no statistical difference of the suPAR expression in effective group or remission group before treated [(257.60±32.47) pg/ml,(331.00±99.80) pg/ml] compated with healthy group (t =2.04,t =1.83,P ＞ 0.05),and that in the invalid group,serum levels of suPAR [(562.20±291.0) pg/ml] was statistically significant with that of the effective group and that of the healthy control group (t =3.92,t =1.93,P ＜ 0.05).Through monitoring 17 patients,the suPAR of patients before treatment [(437.65±131.43) pg/ml] was statistically significant with that of healthy control group (P ＜0.01) and effective group after treatment [(298.76±108.59) pg/ml] (P ＜0.01).Serum suPAR of more than 2-year-survival patients [(333.02±85.37) pg/nl] during initial diagnosis was not statistically difference with that of the healthy control group (t =1.81,P ＞ 0.05).Serum suPAR of less than 2-year-survival patients [(646.01±103.97) pg/ml] was statistically different with that of healthy control group (t =3.84,P ＜0.01) and that of more than 2-year-survival patient group (t =3.50,P ＜0.01).ConclusionThe suPAR value was correlated to the stability of multiple myeloma,and can be used for disease prognosis and estimation of survival time.And the high expression of suPAR was related to the adverse prognosis.