A pathogenic bacterium (S636), identified as Streptococcus iniae, was isolated from turbot (Scophthalrnus maximus) in 2005. We immunized turbot with formalin-killed S. iniae four times (on days 1, 14, 21, and 28...A pathogenic bacterium (S636), identified as Streptococcus iniae, was isolated from turbot (Scophthalrnus maximus) in 2005. We immunized turbot with formalin-killed S. iniae four times (on days 1, 14, 21, and 28) by intraperitoneal inoculation. After each vaccination, we obtained serum samples and isolated the lymphocytes from the peripheral blood, spleen, pronephros, and mesonephros. We measured surface Ig-positive (sIg+) lymphocytes and serum antibody levels from these organs using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively, using monoclonal antibodies against turbot immunoglohulin. We confirmed that the antibody reacted with both the surface and plasma Ig by confocal laser scanning microscopy and electron microscopy. The percentage of sIg+ in the lymphocytes increased following each successive vaccination. The mean percentage increased from 31.96% (control) to 37.49%, 38.36%, 42.9%, and 51.63% in the peripheral blood; from 27.09% to 36.63%, 36.81%, 39.28%, and 46.0% in the spleen; from 22.2% to 28.99%, 29.21%, 32.83%, and 41.58% in pronephros; and from 18.12% to 22.17%, 22.45%, 25.69%, and 31.68% in the mesonephros. The ELISA results were consistent with these results. Both the total and specific antibody levels increased with each vaccination. The mean OD value of the specific antibody assay increased from 0.094, to 0.269, 0.283, 0.333, and 0.421; for total antibody the mean OD value increased from 0.133, to 0.292, 0.323, 0.413, and 0.527.展开更多
We obtain an equation among invariants obtained from the Alexander module of an amphicheiral link. For special cases, it deduces necessary conditions on the Alexander polynomial. By using the present results and some ...We obtain an equation among invariants obtained from the Alexander module of an amphicheiral link. For special cases, it deduces necessary conditions on the Alexander polynomial. By using the present results and some known results, we show that the Alexander polynomial of an algebraically split component- preservingly (±)-amphicheiral link with even components is zero, and we determine prime amphieheiral links with at least 2 components and up to 9 crossings.展开更多
Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)...Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)-myosin-light-chain kinase(MLCK)pathway.Methods:Forty-eight male Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model control(MC)group,an HPM group and a mesalazine(MESA)group,with 12 rats in each group.Trinitrobenzene sulfonic acid(TNBS)was administered to establish CD models.When the model was confirmed a success,the HPM group rats were treated with HPM at Tianshu(ST 25)and Qihai(CV 6),while the MESA group rats were given MESA solution by lavage.When the intervention finished,the colonic epithelial tissues were separated,purified and cultured in each group to establish the intestinal epithelial barrier model in vitro,and TNF-αwas added(100 ng/mL)in the culture medium and maintained for 24 h to establish an increased epithelial permeability model.Transepithelial electrical resistance(TEER)was used to examine the permeability of the barrier;Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway;immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium.Results:After TNF-αinduction,compared with the MC+TNF-αgroup,the TEER value increased significantly in the HPM+TNF-αand MESA+TNF-αgroups(both P<0.001);the expressions of nuclear factor kappa B(NF-κB)p65,MLCK,myosin light chain(MLC),tumor necrosis factor receptor-associated factor 6(TRAF6)and receptor interaction protein-1(RIP1)decreased significantly(P<0.01 or P<0.05),and the expression of zinc finger protein A20(A20)increased significantly(P<0.01);the expressions of occludin,claudin-1,zonula occludens protein 1(ZO-1)and F-actin also increased significantly(all P<0.01).Compared with the MESA+TNF-αgroup,the expressions of MLC,occludin,claudin-1,ZO-1 and F-actin increased significantly in the HPM+TNF-αgroup(P<0.01 or P<0.05).Conclusion:HPM can protect or repair the damage of intestinal epithelial barrier in CD rats,which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.展开更多
基金Supported by the National Basic Research Program of China (973 Program, No 2006CB101806)the High Technology Research and Development Program of China (863 Program,No 2006AA100306)the National Natural Science Fundation of China (No 30771648)
文摘A pathogenic bacterium (S636), identified as Streptococcus iniae, was isolated from turbot (Scophthalrnus maximus) in 2005. We immunized turbot with formalin-killed S. iniae four times (on days 1, 14, 21, and 28) by intraperitoneal inoculation. After each vaccination, we obtained serum samples and isolated the lymphocytes from the peripheral blood, spleen, pronephros, and mesonephros. We measured surface Ig-positive (sIg+) lymphocytes and serum antibody levels from these organs using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively, using monoclonal antibodies against turbot immunoglohulin. We confirmed that the antibody reacted with both the surface and plasma Ig by confocal laser scanning microscopy and electron microscopy. The percentage of sIg+ in the lymphocytes increased following each successive vaccination. The mean percentage increased from 31.96% (control) to 37.49%, 38.36%, 42.9%, and 51.63% in the peripheral blood; from 27.09% to 36.63%, 36.81%, 39.28%, and 46.0% in the spleen; from 22.2% to 28.99%, 29.21%, 32.83%, and 41.58% in pronephros; and from 18.12% to 22.17%, 22.45%, 25.69%, and 31.68% in the mesonephros. The ELISA results were consistent with these results. Both the total and specific antibody levels increased with each vaccination. The mean OD value of the specific antibody assay increased from 0.094, to 0.269, 0.283, 0.333, and 0.421; for total antibody the mean OD value increased from 0.133, to 0.292, 0.323, 0.413, and 0.527.
基金supported by National Natural Science Foundation of China (Grant No. 10801021/a010402)
文摘We obtain an equation among invariants obtained from the Alexander module of an amphicheiral link. For special cases, it deduces necessary conditions on the Alexander polynomial. By using the present results and some known results, we show that the Alexander polynomial of an algebraically split component- preservingly (±)-amphicheiral link with even components is zero, and we determine prime amphieheiral links with at least 2 components and up to 9 crossings.
基金国家自然科学基金项目,No.81674069 and No.81473757973 Program,国家重点基础研究发展计划项目,No.2015CB554500。
文摘Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)-myosin-light-chain kinase(MLCK)pathway.Methods:Forty-eight male Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model control(MC)group,an HPM group and a mesalazine(MESA)group,with 12 rats in each group.Trinitrobenzene sulfonic acid(TNBS)was administered to establish CD models.When the model was confirmed a success,the HPM group rats were treated with HPM at Tianshu(ST 25)and Qihai(CV 6),while the MESA group rats were given MESA solution by lavage.When the intervention finished,the colonic epithelial tissues were separated,purified and cultured in each group to establish the intestinal epithelial barrier model in vitro,and TNF-αwas added(100 ng/mL)in the culture medium and maintained for 24 h to establish an increased epithelial permeability model.Transepithelial electrical resistance(TEER)was used to examine the permeability of the barrier;Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway;immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium.Results:After TNF-αinduction,compared with the MC+TNF-αgroup,the TEER value increased significantly in the HPM+TNF-αand MESA+TNF-αgroups(both P<0.001);the expressions of nuclear factor kappa B(NF-κB)p65,MLCK,myosin light chain(MLC),tumor necrosis factor receptor-associated factor 6(TRAF6)and receptor interaction protein-1(RIP1)decreased significantly(P<0.01 or P<0.05),and the expression of zinc finger protein A20(A20)increased significantly(P<0.01);the expressions of occludin,claudin-1,zonula occludens protein 1(ZO-1)and F-actin also increased significantly(all P<0.01).Compared with the MESA+TNF-αgroup,the expressions of MLC,occludin,claudin-1,ZO-1 and F-actin increased significantly in the HPM+TNF-αgroup(P<0.01 or P<0.05).Conclusion:HPM can protect or repair the damage of intestinal epithelial barrier in CD rats,which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.