Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat ant...Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat anti-mouse CD35 monoclonal antibody and FITC-conjugated goat anti-mouse antibody.Using flow cytometry,we determined the number of ECR1.Using microscope,we studied the adherence between erythrocyte immunity and C3b receptor or tumor-cell by RBC-C3bRR and DTER.Results Comparing the mean value of the number of CR1 on each RBC of high and middle groups with control groups,the mean value of the number of CR1,RBC-C3bRR and DTER of Asparagus officinalis polysaccharide groups are increased significantly.Conclusions Asparagus officinalis polysaccharide can improve the erythrocyte function of S180 mice,which may be one of its most important antitumor mechanisms.展开更多
Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated pro...Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated protein kinases (MAPKs, including ERK, JNK, and p38MAPK), and to highlight the molecular mechanism of CD 4 + cell deletion in AIDS Methods HOS cells (HOS CR2) and HOS CD4 cells (HOS CD4CR2) stably expressing CR2 were established and then identified by FACS and Western blotting Activation and blocking tests of MAPKs were assessed by Western blot Cell proliferation was determined using Cell Titer 96 Aqueous One Solution Reagent Results FACS results showed that the positive rates of HOS CR2 and HOS CD4CR2 cells were greater than 96%, and Western blot showed that the CR2 expression levels on HOS CR2 and HOS CD4CR2 cells were high Activation and blocking tests of MAPKs (ERK, JNK, and p38MAPK) were carried out in HOS CR2, HOS CD4, and HOS CD4CR2 cells The activation of MAPKs in HOS CR2 cells stimulated with PMA (100 ng/ml) and NHS (10%) was identical The activation of MAPKs increased at 5 minutes, reached a peak at 10 minutes, and decreased to baseline within 30 minutes, all in a time dependent manner; the activation of MAPKs was blocked by anti CR2 McAb, PD98059 (inhibitor of ERK), and Wortmanin (inhibitor of PI 3K), respectively In HOS CD4 cells, MAPKs were activated by HIV gp160 In HOS CD4CR2 cells, MAPK activation was induced by HIV gp160, 10% NHS, and HIV gp160+10%NHS; phosphorylation of p38MAPK was dramatically induced by HIV gp160+NHS, and lasted for 1 hour The cell proliferation results showed that HIV gp160 inhibited the proliferation of HOS CD4 and HOS CD4CR2 cells ( P <0 01) and that NHS enhanced the effect of HIV gp160 ( P <0 01) Conclusions The activation of MAPKs is independently mediated by CR2 and that anti CR2 McAb, PD98059, and Wortmanin block the activation of MAPKs, respectively The results of the signal transduction and cell proliferation assays of HOS CD4CR2 cells show that CR2 plays a role in the pathogenesis of HIV infection, especially in the inhibition of CD 4 + cell proliferation展开更多
目的观察心肾综合征(CRS)患者服用小剂量辛伐他汀后血清补体C3、C4水平及外周血单核细胞Ⅰ型(CR1)、Ⅱ型(CR2)补体受体m RNA表达的变化。方法随机入选2012年1~12月在南京市胸科医院门诊和病房住院的CRS患者56例,根据NYHA心功能分级进行...目的观察心肾综合征(CRS)患者服用小剂量辛伐他汀后血清补体C3、C4水平及外周血单核细胞Ⅰ型(CR1)、Ⅱ型(CR2)补体受体m RNA表达的变化。方法随机入选2012年1~12月在南京市胸科医院门诊和病房住院的CRS患者56例,根据NYHA心功能分级进行心功能分级,根据简化的MDRD公式计算估计的肾小球滤过率。入选对象每晚睡前服用辛伐他汀10 mg,连续服用8周。入选后次日清晨及治疗第8周清晨抽取空腹静脉血,全自动生化分析仪测定血清C3、C4水平;采集外周静脉血提取外周血单个核细胞,提取总RNA并行RT-PCR检测CR1及CR2 m RNA的表达。结果 CRS患者血清C3水平治疗后较治疗前降低[(1.14±0.35)和(1.22±0.31)g/L,P<0.05],外周血单核细胞CR1 m RNA治疗后较治疗前表达增强(0.76±0.10和0.70±0.11,P<0.05)。结论 CRS患者服用小剂量辛伐他汀后,血清C3水平降低及外周血单核细胞CR1核酸表达上调可能共同参与患者心肾功能的改善。展开更多
目的实现人补体受体1型(complement receptor type 1,CR1)活性片段SCR15-18在毕赤酵母中的分泌表达。方法用PCR从原核表达质粒pET32a-sCR1-SCR15-18上扩增CR1-SCR15-18的编码基因,并克隆到毕赤酵母分泌表达质粒pPIC9,构建重组质粒pPIC9-...目的实现人补体受体1型(complement receptor type 1,CR1)活性片段SCR15-18在毕赤酵母中的分泌表达。方法用PCR从原核表达质粒pET32a-sCR1-SCR15-18上扩增CR1-SCR15-18的编码基因,并克隆到毕赤酵母分泌表达质粒pPIC9,构建重组质粒pPIC9-CR1-SCR15-18,鉴定后测序;重组质粒电转化整合到毕赤酵母GS115基因组中,菌落PCR技术筛选阳性转化株;经摇瓶发酵和甲醇诱导,SDS-PAGE和Western blot鉴定目的蛋白的表达;Ni2+-NTA agarose亲和层析纯化目的蛋白,并用体外抑制补体溶血反应实验测定目的蛋白的生物活性。结果成功构建重组表达质粒pPIC9-CR1-SCR15-18;SDS-PAGE和Western blot证实目的基因在毕赤酵母中成功分泌表达;表达产物经镍柱快速纯化后,能够明显抑制补体的体外溶血。结论在毕赤酵母中成功实现了CR1-SCR15-18蛋白的分泌表达,该蛋白具有较高的抑制补体溶血的生物活性。展开更多
文摘Objective To study the effect of Asparagus officinalis polysaccharide on the number and activity of erythrocyte complement receptor 1 in S180 mice.Methods Red blood cells from mice venous blood were labeled by rat anti-mouse CD35 monoclonal antibody and FITC-conjugated goat anti-mouse antibody.Using flow cytometry,we determined the number of ECR1.Using microscope,we studied the adherence between erythrocyte immunity and C3b receptor or tumor-cell by RBC-C3bRR and DTER.Results Comparing the mean value of the number of CR1 on each RBC of high and middle groups with control groups,the mean value of the number of CR1,RBC-C3bRR and DTER of Asparagus officinalis polysaccharide groups are increased significantly.Conclusions Asparagus officinalis polysaccharide can improve the erythrocyte function of S180 mice,which may be one of its most important antitumor mechanisms.
文摘Background Complement receptor type 2 (CR2) is the receptor for C3d and C3dg and for Epstein Barr virus The aim of our study was to explore whether CR2 can independently mediate the activation of mitogen activated protein kinases (MAPKs, including ERK, JNK, and p38MAPK), and to highlight the molecular mechanism of CD 4 + cell deletion in AIDS Methods HOS cells (HOS CR2) and HOS CD4 cells (HOS CD4CR2) stably expressing CR2 were established and then identified by FACS and Western blotting Activation and blocking tests of MAPKs were assessed by Western blot Cell proliferation was determined using Cell Titer 96 Aqueous One Solution Reagent Results FACS results showed that the positive rates of HOS CR2 and HOS CD4CR2 cells were greater than 96%, and Western blot showed that the CR2 expression levels on HOS CR2 and HOS CD4CR2 cells were high Activation and blocking tests of MAPKs (ERK, JNK, and p38MAPK) were carried out in HOS CR2, HOS CD4, and HOS CD4CR2 cells The activation of MAPKs in HOS CR2 cells stimulated with PMA (100 ng/ml) and NHS (10%) was identical The activation of MAPKs increased at 5 minutes, reached a peak at 10 minutes, and decreased to baseline within 30 minutes, all in a time dependent manner; the activation of MAPKs was blocked by anti CR2 McAb, PD98059 (inhibitor of ERK), and Wortmanin (inhibitor of PI 3K), respectively In HOS CD4 cells, MAPKs were activated by HIV gp160 In HOS CD4CR2 cells, MAPK activation was induced by HIV gp160, 10% NHS, and HIV gp160+10%NHS; phosphorylation of p38MAPK was dramatically induced by HIV gp160+NHS, and lasted for 1 hour The cell proliferation results showed that HIV gp160 inhibited the proliferation of HOS CD4 and HOS CD4CR2 cells ( P <0 01) and that NHS enhanced the effect of HIV gp160 ( P <0 01) Conclusions The activation of MAPKs is independently mediated by CR2 and that anti CR2 McAb, PD98059, and Wortmanin block the activation of MAPKs, respectively The results of the signal transduction and cell proliferation assays of HOS CD4CR2 cells show that CR2 plays a role in the pathogenesis of HIV infection, especially in the inhibition of CD 4 + cell proliferation
文摘目的观察心肾综合征(CRS)患者服用小剂量辛伐他汀后血清补体C3、C4水平及外周血单核细胞Ⅰ型(CR1)、Ⅱ型(CR2)补体受体m RNA表达的变化。方法随机入选2012年1~12月在南京市胸科医院门诊和病房住院的CRS患者56例,根据NYHA心功能分级进行心功能分级,根据简化的MDRD公式计算估计的肾小球滤过率。入选对象每晚睡前服用辛伐他汀10 mg,连续服用8周。入选后次日清晨及治疗第8周清晨抽取空腹静脉血,全自动生化分析仪测定血清C3、C4水平;采集外周静脉血提取外周血单个核细胞,提取总RNA并行RT-PCR检测CR1及CR2 m RNA的表达。结果 CRS患者血清C3水平治疗后较治疗前降低[(1.14±0.35)和(1.22±0.31)g/L,P<0.05],外周血单核细胞CR1 m RNA治疗后较治疗前表达增强(0.76±0.10和0.70±0.11,P<0.05)。结论 CRS患者服用小剂量辛伐他汀后,血清C3水平降低及外周血单核细胞CR1核酸表达上调可能共同参与患者心肾功能的改善。
