Objective To dynamically observe changes of IgG, its subclasses and IgE in sera of mice by immunization with mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis (Era). Methods Forty Balb/c mice of 12-14 week old and 20-25 g weight were intranasally vaccinated by the vaccine, 4 mice were killed randomly by the weight on 0,2,4,6,8,10,12,14,16 and 18 weeks of immunization respectively, sera were gathered from the eyeball to measure IgG, its subclasses and IgE by routine ELISA. Results Levels of IgG, IgG2a and IgG2b in the sera of mice increased obviously on 2-18 weeks, reached the highest level on 10, 4 and 4 weeks respectively, the value was 0.095±0.033,0.022±0.001,0.023±0.003 respectively, as compared with the value on 0 week(0.030±0.013,0.012±0.004,0.013±0.004), the difference being statistically significant(q=2.95,4.87,2.81 respectively, P < 0.01 or P < 0.05); levels of IgG1, IgG3 and IgE in the sera of mice decreased remarkably on 2-18 weeks,came to the lowest level on 4,2,6 weeks respectively, the value was 0.031±0.004,0.136±0.002,0.114±0.002 respectively, as compared with the value on 0 week(0.192±0.007, 0.175±0.013,0.024±0.003), the difference being statistically significant (q =5.16,4.93,5.32 respectively, P < 0.01 or P < 0.05). Conclusion Helper T cell(TH) Ⅰ response is induced in mice by mixed recombinant of BCG-Em Ⅱ/3 and BCG-Em14-3-3 vaccine on early immunization.

Objective To dynamically observe the changes of subsets of splenocytes in mice by immunization with recombinant BCG-Eg95 vaccine of Echinococcus granulosus(Eg).Methods Balb/c mice were divided randomly into 3 groups according to their weishts:intranasal group.per os group and PBS control.The mice were vaccinated intranasally or orally by the vaccine respectively in experimental groups.and the control mice were given phosphate buffer saline intranasally.These mice were killed to get spleen on 0,2,4,6,8,10,12, 14,16 and 18 week of immunization,respectively.Splenocytes were separated to measure subsets of CD4+ and CD8+T ceUs by FACsort.Results There were marked differences in ratio of CD4+ and CD8+ subsets among the different groups(F value were 21.56 and 22.08 respectively,P＜0.05).There were very marked differences in ratio of CD4+ and CD8+ subsets in different weeks(F value were 5.75 and 6.29 respectively.P＜0.01).In the intranasal group,CD4+ and CD8+ T subsets increased obviously in 6～18 weeks and 12 weeks,and reached the highest level on 10 and 12 week,espectively.Their values were 0.348±0.013 and 0.090±0.003.respectively.There were marked or very marked differences(q value were 7.32 and 5.32 respectively,P＜0.01 or＜0.05)in comparison with 0 week(0.230±0.022 and 0.069±0.015).In the oral group,CD4+and CD8+ subsets rose reinarkablv on 6-16 weeks and 8-18 weeks,achieved the hishest level on 10 and 16 weeks,respectively.Their vahes were 0.405± 0.006 and 0.096±0.004,respectively.There were marked or very marked difference(q value were 7.53 and 5.35 respectively,P＜0.01 or＜0.05)in comparison with week 0(0.230±0.022 and 0.069±0.015).Conclusion CD4+and CD8+T subsets may play an important role in immune response induced in mice by rBCG-Eg95 vaccine.

Objective To investigate the changes of cytokines of splenocytes in mice immunized with recombinant BCG-Em Ⅱ/3 vaccine of Echinococcus multilocularis(Em)and consequently challenged with Em protoscoleces.Methods Balb/c mice were subcutaneously or intranasally vaccinated and challenged with Em were separated and cultured with EmAg,ConA or PHA,respectively.The supematants were gathered to measure the levels of IL-2,IFN-γ,TNF-α.and IL-4 by ELISA Kits.Results The levels of IL-2,IFN-γ,TNF-α and IL-4 in the subcutaneous group were(34.6±2.7),(34.5±2.8),(265.0 ±0.0)and(9.8±2.6)ng/L respeetively:those in the intranasal group were(32.5±2.2),(33.6±2.7),(130.0±0.0)and(10.4±27)ng/L respectively;those in the control were(25.0±1.9),(30.0±0.0),(10.0±0.0)and(12.5±2.7)ng/L,respectively:there were statistical differences between the immunized groups and control group(P＜0.01 or＜0.05);The level of TNF-α in the subcutaneous group was higher than that in the intranasal group.Conclusion Th1 response has been induced in mice vaccinated with rBCG-Em Ⅱ/3 vaccine and challenged with Em protoscoleces.

This paper mainly discusses the purpose , content , organization of the curriculum design of fermentation engineering and equipment and the teching importance to bioengineering major.

Objective To measure the temperature sensation threshold of trunk skin in healthy adults. Methods The threshold of cold sensation, warm sensation, cold pain sensation and heat pain sensation of trunk skin key points (T3, T7 and T11) were measured with Thermal Sensory Analyzer in 123 healthy adults. Results The thresholds of cold, warm, cold pain and heat pain sensations were obtained. The standard deviation of cold and warm threshold was less than that of heat pain. The range of cold sensation threshold was the largest. The heat pain sensation threshold increased with segmental declining and the sensation threshold increased with age. Conclusion Normal reference value should be established variously with the segment and age. The threshold of cold, warm varies less, while the threshold of cold pain and heat pain varies too much.

Objective To detect the expression levels of interleukin (IL)-32 and tumor necrosis factor (TNF)-α in peripheral blood of rheumatoid arthritis (RA) patients,and analyze the relations between IL-32 and level of erythrocytes (ESR),creactive protein (CRP),rheumatoid factor (RF),the disease activity index 28 (DAS28) scores in RA patients.The aim of this study is to identify the clinical significance of IL-32 in RA patient.Methods We collected 97 specimens of peripheral blood from RA patients,including 36 RA patients with high active disease,33 RA patients with moderate active disease,28 RA patients with stable disease,36 non-RA connective tissue diseases (CTD) patients and 30 healthy volunteers.The mRNA levels of IL-32,TNF-α in the peripheral blood mononuclear cells (PBMC) were determined by quantitative real-time PCR (QRT-PCR).The concentration of IL-32,TNF-αt in plasma were detected by enzyme linked immunosorbent assay (ELISA),one-way AN OVA and Spearman's corrclation and multiple linear regression were adopted for statistical analysis.Results ①The mRNA levels of IL-32 and TNF-α in active RA patients were higher than those in healthy control group (0.29±0.07,0.24±0.06 vs 0.11±0.06 for IL-32 in the high active RA group and moderate active RA group versus the healthy group; and 0.042±0.022,0.012±0.006 vs 0.004±0.002 for TNF-α in the high active RA group and moderate active RA group versus the healthy group,P＜0.01),and than those in stable RA patients,and non-RA CTD patients.There was no significant difference among the stable disease groups,healthy control group and non-RA CTD group in the level of mRNA.② The concentration of IL-32,TNF-α in the plasma of RA patients with active disease was higher than that of healthy control group [(107±42) pg/ml,(66±27) pg/ml vs (20±10) pg/ml for IL-32 in the high active RA group and moderate active RA group versus the healthy group; and (261±77) pg/ml,(186±84) pg/ml vs (72±20) pg/ml for TNF-αt in the high active RA group and moderate active RA group versus the healthy group,P＜0.01].③ The expression levels of IL-32 was positively correlated with the level of TNF-α,ESR,CRP,RF,DAS28 scores in the plasma of RA patients (r=0.568,0.321,0.185,0.325,0.303 respectively,P＜0.01).④ The expression levels of IL-32 was positively correlated with the level of TNF-α (the regression coefficients and P values were b=1.721,P＜0.01 and b=0.176,P＜0.01 respectively).Conclusion The levels of IL-32 in peripheral blood may be used as an indicator for disease activity of RA.

Background The occurrence of experimental myopia may be related to glucagon,and within the range of certain concentration,glucagon may inhibit the development of myopia,but its exact action mechanism is not completely clear.Objective Purpose of this study was to evaluate the effects of glucagon on the proliferation of guinea pig scleral fibroblast cells(GSFCs) and the possible role of glucagon in myopic scleral remodeling.Methods The scleral tissue was obtaincd from the clean blooded guinea pig aged 15 days.GSFCs were cultured and identified with vimentin antibody,cytokeratin antibody and S-100 antibody.0,5,10,50,100,200 μg/L glucagon was added into the different cultured hole for 24 hours respectively,and the growth and proliferation (A490 value) of GSFCs was detected by MTT colorimetric assay.Then the A490 value of GSFCs was assayed in 1 day,2,3,5,7 days under the 50 μg/L glucagon action.Matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2)levels(A450 value)in GSFCs were detected by enzyme-linked immuno sorbent assay (ELISA)72 hours after the cells cultured.Results Passaged GSFCs showed the dendric array at lower density and gyrate array at the higher density with the positive response for vimentin.A490 values of GSFCs were gradually increased with the rise of glucagon concentration(F=32.340,P=0.013).When the glucagon concentration was 10-200 μg/L,the A490Value of GSFCs was higher than that without glucagon group,showing signifitant differences between them(t =5.575,6.627,16.074,12.003,P＜0.05).Under the 50 μg/L glucagon action,A490 values were significantly accented with the time lapse (Ftime =10.610,P =0.024),and the A490 values also were significantly higher than the parallel control groups without glucagon(Fgroup =9.068,P=0.039).MMP-2 level was gradually declined with the enhance of glucagon within range of 5-200 μg/L(F=153.639,P=0.036),but no significant difference was found in TIMP-2 expression(F=24.770,P=3.250).Conclusions Glucagon can promote the proliferation of GSFCs in vitro,and the synthesis of MMP-2shows a concentration-and time-dependent manner.

Objective To compare the therapeutic effect of the combination of ~(99m)Tc-methnetium methylene diphosphunate and colloidal chromic,phosphate ~(32)p with the monotherapies in rats with adjuvant arthritis(AA).Methods In this study,50 Sprague-Dawley rats were randomly divided into five groups with 10 rats in each:normal control group(arthritis induced and treated with the same volume of normal saline via the same administration route),AA control group(arthritis induced with adjuvant and treated with normal sa- line),~(32)p colloid group(arthritis induced with adjuvant and treated with single intra-articular injection of col- luidal chromic phosphate ~(32)p 0.74 MBq and intra-peritoneal injection of normal saline every other clay),~(99m)Tc- MDP group(arthritis induced with adjuvant and treated with intra-peritoneal injection of ~(99m)Tc-MDP 2.5?10~(-3)?g/kg every other day and single intra-articular injection of normal saline),combination group(arthritis in- duced with adjuvant and treated with the combination of the two drugs).The diameter of the left hind ankle, serum levels of turnout necrosis factor(TNF)and interleukin-1?(IL-1?),and the pathological changes of an- kle joints were tested at different time points.Results The diameter of the left hind ankle at week 4 was smaller in the combination group than that of the ~(32)p colloid treatment alone group [(7.11?0.34)mm vs(7.57?0.29)mm,P

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