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AIM:To establish the monoclonal antibody against human B lymphocyte stimulator (hBLyS) by DNA immunization and analyse its characterization. METHODS:The 858 bp DNA fragment of hBLyS was cloned into pcDNA3 plasmids. The cloned insert was identified by both sequence analysis and double digestion of the recombinant plasmid with restriction enzymesXho Iand EcoR I. After the splenocytes from BALB/c mice immunized with the recombinant plasmid of pcDNA3/hBLyS were fused with myeloma cells SP2/0,the hybridoma which can produce monoclonal antibodies against hBLyS were obtained. The specificity of anti-BLyS monoclonal antibody from hybridoma was verified by ELISA, Western blot and flow cytometry. RESULTS:The recombinant mammalian cell expression vector of pcDNA3/hBLyS was constructed,the sequence of the insert gene was identified to be the sequence encoding hBLy S antigen. The culture supernatants of hybridoma 9c10 were tested to be the monoclonal antibody with specificity against hBLyS on human peripheral blood CD3+T cell activated by hIFN-γ by ELISA,Western blot and flow cytometry.CONCLUSION:The monoclonal antibodies against hBLyS with high activity and specificity have been established successfully, and will be an useful tool in the studies of relationship between hBLyS and human autoimmunity diseases. 相似文献
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AIM: To observe the effect of Jiawei Sini Decoction (JWSND) on glucocorticoid receptor (GCR) in thymocytes of chronic psychological stress rats. METHODS: The rats were randomly divided into 4 groups, control group (C), model group (M), group treated by JWSND C1, group treated by ginsenosides C2. The number of thymocyte GCR sites and the GCR nuclear thanslocation rate were detected by radioimmunoassay. RESULTS: Compared with the control group, in the model group, the thymocyte weight index lowered significantly ( P<0.05 ), and the GCR nuclear thanslocation rate was increased significantly ( P<0.01 ), but the number of thymocyte GCR sites was unchanged. Compared with the model group, thymus gland weight indexes of C1 and C2 were increased significantly ( P<0.05 ), while the GCR nuclear thanslocation rate lowered significantly ( P<0.01 ). Moreover, no significant difference was found in all indexes between C1 and control group. CONCLUSION: The inhibitory effect of glucocorticoid on the thymus could be significantly reversed by JWSND via suppressing the thanslocation of GCR from cytoplasm to nucleus in chronic psychological stress rats. 相似文献
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中生菌素对水稻白叶枯病的防治机制 总被引:6,自引:1,他引:6
对不同白叶枯病抗性水稻品种用200μg/ml中生菌素55℃温汤药液浸种,自然降温,秧田3—4叶期和移栽前5d各用30μg/ml中生菌素处理后,于成株期剪叶接种白叶枯病菌。结果表明中生菌素前期处理,于成株期接种白叶枯病菌时,高抗、中等抗性和感病品种中过氧化物酶、苯丙氨酸解氨酶和多酚氧化酶3种酶活性都有不同程度的提高,且以中等抗性品种酶活提高最多,接种24和48h,3种酶活性较清水对照分别增加26.92%、26.74%、24.06%和7.09%、1.31%、1.60%。盆栽试验表明,中生菌素对中抗品种的防治效果最好,达58.4%。说明中生菌素对水稻防御酶活性的诱激作用是其防治白叶枯病的机制之一。 相似文献
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XU Ruo-bing WEN Jian-ming ZHANG Meng LV Chang-hai XIAO Gang ZHANG Wen-min LIANG Hui-zhen 《园艺学报》2004,20(11):1982-1988
AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT. 相似文献
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CHEN Chun-yan JIA Ji-hui PAN Xiang-lin ZHANG Qi WANG Juan-dong ZHOU Ya-bin 《园艺学报》2004,20(7):1183-1186
AIM: To study the signal transduction pathway of apoptosis initiation induced by homoharringtonine in HL-60 cells. METHODS: After establishing the model of apoptosis initiation induced by homoharringtonine in HL-60 cells, at the point of apoptosis initiation, molecular caspase-3, Bcl-2, Bax and Fas/FasL were measured with flow cytometry and transmission electron microscope. ERK2 and P38 expression in HL-60 cells were detected by using immunohistochemistry. RESULTS: The model of apoptosis initiation induced by homoharringtonine was established in HL-60 cells. At the point of apoptosis initiation, upregulation of caspase-3 and decrease in Bcl-2/Bax were observed. However, the expression of Fas/FasL did not significantly change. ERK2 expression decreased and P38 expression increased. CONCLUSIONS: Caspase-3, Bcl-2, Bax and mitogen activated protein kinase pathways were involved in signal transduction of apoptosis initiation induced by homoharringtonine in HL-60 cells. 相似文献