目的:探讨胰岛素样生长因子-1(IGF-1)对脓毒症大鼠血浆应激激素、肝和骨骼肌胰岛素受体(InsRα、β)蛋白及mRNA表达的影响。方法:将SD大鼠采用盲肠结扎穿孔法(CLP)建立腹腔感染及颈静脉置管肠外营养(PN)模型后,随机分为脓毒症组(n=10)...目的:探讨胰岛素样生长因子-1(IGF-1)对脓毒症大鼠血浆应激激素、肝和骨骼肌胰岛素受体(InsRα、β)蛋白及mRNA表达的影响。方法:将SD大鼠采用盲肠结扎穿孔法(CLP)建立腹腔感染及颈静脉置管肠外营养(PN)模型后,随机分为脓毒症组(n=10)、PN组(n=10)及IGF-1组(n=10),另设正常组(n=10)。正常组和脓毒症组给予等渗盐水,PN组给予营养支持,IGF-1组为PN+IGF-1静脉注射,连续5 d。第6 d行颈动脉置管,剖腹取门静脉血测定血糖、胰岛素、胰高血糖素和IGF-1;免疫组化法和Real-Tim e PCR法测定肝、骨骼肌组织InsRα、β蛋白及mRNA表达。结果:脓毒症大鼠血浆葡萄糖、胰岛素、胰高血糖素明显升高,IGF-1明显下降(P<0.01)。IGF-1组前三者较脓毒症组和PN组明显降低,IGF-1则明显升高(P<0.05)。脓毒症组大鼠肝及骨骼肌InsRα、β蛋白及mRNA均下降,IGF-1组较脓毒症组和PN组明显升高(P<0.01)。结论:IGF-1可改善应激激素之间的平衡,明显增加脓毒症大鼠InsRα、β蛋白水平和mRNA的表达,降低血糖水平。展开更多
Methylene blue injection has a stronger direct inactivation on VZV in vitro. When the injection was diluted from 116 to 1128, which was obvious (P<001 and P<005). The MIC was 1222, the IC50 was 1135 and IC90 was...Methylene blue injection has a stronger direct inactivation on VZV in vitro. When the injection was diluted from 116 to 1128, which was obvious (P<001 and P<005). The MIC was 1222, the IC50 was 1135 and IC90 was 177. The results of microcellculture method showed when the injection was diluted from 116 to 164, it also effectively inhibited the proliferation of VZV in WISH continuous celllines (P<001 and P<005). The MIC was 195, the IC50 was 145 and the IC90 was 121.展开更多
The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two ci...The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on NIH3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas + Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8.5×10 5 colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53.81±6.9 %), and significantly induced the apoptosis of Fas + Yac-1 cells (56.78±4.5 %), as both were cocultured for 5 h at 1∶1 ratio, whereas it is 7.08±3.4 % in control group (P<0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.展开更多
文摘目的:探讨胰岛素样生长因子-1(IGF-1)对脓毒症大鼠血浆应激激素、肝和骨骼肌胰岛素受体(InsRα、β)蛋白及mRNA表达的影响。方法:将SD大鼠采用盲肠结扎穿孔法(CLP)建立腹腔感染及颈静脉置管肠外营养(PN)模型后,随机分为脓毒症组(n=10)、PN组(n=10)及IGF-1组(n=10),另设正常组(n=10)。正常组和脓毒症组给予等渗盐水,PN组给予营养支持,IGF-1组为PN+IGF-1静脉注射,连续5 d。第6 d行颈动脉置管,剖腹取门静脉血测定血糖、胰岛素、胰高血糖素和IGF-1;免疫组化法和Real-Tim e PCR法测定肝、骨骼肌组织InsRα、β蛋白及mRNA表达。结果:脓毒症大鼠血浆葡萄糖、胰岛素、胰高血糖素明显升高,IGF-1明显下降(P<0.01)。IGF-1组前三者较脓毒症组和PN组明显降低,IGF-1则明显升高(P<0.05)。脓毒症组大鼠肝及骨骼肌InsRα、β蛋白及mRNA均下降,IGF-1组较脓毒症组和PN组明显升高(P<0.01)。结论:IGF-1可改善应激激素之间的平衡,明显增加脓毒症大鼠InsRα、β蛋白水平和mRNA的表达,降低血糖水平。
文摘Methylene blue injection has a stronger direct inactivation on VZV in vitro. When the injection was diluted from 116 to 1128, which was obvious (P<001 and P<005). The MIC was 1222, the IC50 was 1135 and IC90 was 177. The results of microcellculture method showed when the injection was diluted from 116 to 164, it also effectively inhibited the proliferation of VZV in WISH continuous celllines (P<001 and P<005). The MIC was 195, the IC50 was 145 and the IC90 was 121.
基金This project was supported by a grant from National Natural Sciences Foundation of China(No.39770 76 7)
文摘The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on NIH3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas + Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8.5×10 5 colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53.81±6.9 %), and significantly induced the apoptosis of Fas + Yac-1 cells (56.78±4.5 %), as both were cocultured for 5 h at 1∶1 ratio, whereas it is 7.08±3.4 % in control group (P<0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.
