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Objective:To establish a method to determine vitamin B 6 and fluocinonide in compound vitamin B 6 gel.Methods:The contents of vitamin B6 and fluocinonide in the gel were determined by HPLC with a Lichrospher CN C 18column (250 mm ×4.6 mm, 5μm).The mobile phase was heptane sulfonate solution (adding 6.8 g potassium phosphate monobasic and 1.0 g heptane sulfonate into 1000 ml water) -methanol –triethylamine (35:65:0.2,adjusting pH to 3.2 with phosphoric acid).The detection wavelength was 238 nm and the column temperature was 25℃.The injection volume was 20 μl.Results: Vitamin B6 had a good linear relationship within the range of 15.0-300.0 μg· ml-1(r=1.000 0), and the average recovery was 99.65% (RSD=0.3, n=9).Fluocinonide had a good linear relationship within the range of 0.4-8.0 μg· ml-1 (r=0.9990), and the average recovery was 99.35% (RSD=0.85, n=9).Conclusion:The method is simple and reproducible, and the result is accurate.The method can be used for the deter-mination of the gel.
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Objective To study the protective effects of garlic polysaecharide on L02 from oxidative injury.Methods Cultured L02 were injured by ethanol.Various concentrations of GP(10、20、40、80 mg/L) were added into culture medium.Then the cellular MDA,SOD,and GSH-Px were determined in order to observe the protection of curcumin and the time-dose-response effects.Meanwhile,HO-1 mRNA was detected by RT-PCR method after ethanol exposure.The expressions of HO- 1 proteins were detected by Western blotling.Results GP (10、20、40、80 mg/L) could reduce oxidative injury induced by ethanol in L02 cells.Liver cells were 100 mmol/L alcohol after 8h exposure,SOD[(3.65±0.42) NU/mg,(4.11±0.16) NU/mg,(4.61 ±0.23)NU/mg],GSH-Px [ (75.96 ± 8.96) mg/mg,(81.83±5.70) mg/mg,(89.32±6.35) mg/mg respectively],GSH-Px[(75.96±8.96),(81.83±5.70),(89.32±6.35) respectively]activity,MDA[(1.05±0.16) nmol/mg,(0.99± 0.12) nmol/mg; (0.78± 0.11) nmol/mg respectively]levels compared with the control group [ (2.35 ±0.28) NU/mg,(54.41 ±8.17) mg/mg,(1.58±0.23) nmol/mg respectively],there was a significant difference (P＜ 0.05 ); HO-1 mRNA expression was in a concentration- dependent effect.Conclusion GP had protective effects on L02 from oxidative injury probably by reducing GSH consumption,improving antioxidant enzyme activity and inhibiting lipid peroxidation reaction at dose-dependent manner.GP could promote expression of HO-1 mRNA co-coordinating role in protecting liver cells from oxidative injury.
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Objective To compare a potential role of dendritic cells (DCs) and macrophages in inducing protective immunity against infection with Schistosoma japonicum. Methods DCs and macrophages were pulsed in vitro with soluble egg antigen (SEA) of S. japonicum. BALB/c mice were injected three times with DCs or macrophages, either antigen-pulsed or not,and challenged with 40 ± 2 cercariae of S. japonicum per mouse. Worms were collected 42 days later by portal perfusion of the mice and egg number of liver was calculated. To evaluate whether protective immunity had been induced by preparations of DCs or macrophages, the worm burden and fertility ( eggs per female per mouse liver) were compared between the groups of mice. The antibody level against SEA was detected by ELISA. Results With respect to mice injected with untreated cells, numbers of worms and eggs per female worms were significantly reduced in the groups of mice having received pulsed DCs (26. 3% and 37.9%, respectively), or pulsed macrophages (22. 0% and 30.7%). Untreated DCs and macrophages induced no significant effects. The antibody level against SEA rose in sera of all groups of mice up to 42 days after the challenge, but most pronounced in those immunized with pulsed DCs, although this was not significantly different from other groups. Conclusion The results suggest that the protective immunity against S. japonicum might be induced by DCs to a higher extent than by macrophages after in vitro pulsing with egg antigen.
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Schistosoma japonicum recombinant antigens were used to immunize mice in order to ob-serve their protective immunity against challenge. It was assumed that the recombinant anti-gens could reduce worm burden (29. 2%- 51. 0%) and inibit the fecundity of female schisto-some (52. 1% - 74. 3%). The antibody titers increased significant 3 weeks after the first im-munization and remained high lever. The results indicated that the recombinant antigens ex-pressed in the E. coli could induce protective immunity against challenge. It might be pro-duced in large scale as an important candidate component of a multivalent vaccine against schistosomiasis japonicum.
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Objective To explore the mechanism of protective immunity against Schistosoma japonicum infection induced by Sj26 gene transfected dendritic cell(DC).Methods 48 BALB/c mice were divided randomly into 4 groups with 12 each.The mice were injected through auricle for three times with Sj26 gene transfected DC(Group A),pcDNA3 transfected DC(Group B),untreated DC(Group C) and RPMI-1640(Group D) respectively,and challenged with 40?2 cercariae of S.japonicum per mouse 2 weeks after the last immunization.Sera from mice were examined for IgG antibody,IFN-? and IL-4 by ELISA.Western blot was used for detecting specific anti-Sj26 IgG antibody.The production of IFN-? and IL-4 in the supernatant of spleen cells stimulated with soluble egg antigen(SEA) and ConA was quantified by sandwich ABC-ELISA.The proliferation of spleen cells were measured with MTT method.Results IgG antibody increased significantly in the mice of group A at 2 weeks after the last immunization(absorbency A491=0.117),higher than that of group B(A491=0.061) and group C(A491=0.058)(P