Based on butterfly microfluidic gene chip technology,a method for rapid,accurate and efficient detection of 19 common pathogenic microorganisms of nosocomial infection was established,and a butterfly microfluidic gene...Based on butterfly microfluidic gene chip technology,a method for rapid,accurate and efficient detection of 19 common pathogenic microorganisms of nosocomial infection was established,and a butterfly microfluidic gene chip with high-throughput detection was designed and fabricated.Using constant temperature amplification technology,using the polymerase with chain replacement function to react at constant temperature(65℃)and combined with microfluidic chip technology,primers were designed according to the target genes of 19 pathogenic microorganisms,and a butterfly microfluidic gene chip which can detect 19 common pathogenic microorganisms of nosocomial infection was made to simplify the inspection operation process and verify the sensitivity of the chip.The butterfly microfluidic gene chip can be used for the rapid and efficient detection of 19 common pathogenic microorganisms of nosocomial infection,and provides a new idea for the detection and auxiliary diagnosis of pathogenic microorganisms of nosocomial infection.展开更多
目的探讨高三尖杉酯碱(HHT)对HT29人结直肠肿瘤细胞增殖和凋亡调控的分子机制。方法用(0、0.0078125、0.015625、0.03125、0.0625、0.125、0.25、0.5、1、2、4、8)μg/m L HHT分别处理HT29细胞24、48、72 h。CCK-8法检测HT29细胞的增殖...目的探讨高三尖杉酯碱(HHT)对HT29人结直肠肿瘤细胞增殖和凋亡调控的分子机制。方法用(0、0.0078125、0.015625、0.03125、0.0625、0.125、0.25、0.5、1、2、4、8)μg/m L HHT分别处理HT29细胞24、48、72 h。CCK-8法检测HT29细胞的增殖,集落形成实验检测细胞的集落形成能力;Hoechst33258染色观察染色质凝集情况,流式细胞术检测细胞凋亡;实时定量PCR检测BAX、Bcl2、胱天蛋白酶3(caspase-3)、caspase-9、哺乳动物雷帕霉素靶蛋白(mTOR)、磷脂酰肌醇3激酶(PI3K)、丙酮酸脱氢酶激酶(PDK1)、蛋白激酶B(AKT)、雷帕霉素靶蛋白调节相关蛋白(raptor)、雷帕霉素不敏感的伴侣蛋白(rictor)的mRNA水平;Western blot法检测BAX、Bcl2、胱天蛋白酶3前体(pro-caspase-3)、裂解型caspase-3(c-caspase-3)、pro-caspase-9、裂解型caspase-9(c-caspase-9)、多腺苷二磷酸核糖聚合酶(PARP)、裂解型PARP(c-PARP)、mTOR、raptor、rictor、PI3K、PDK1、AKT、p-AKT的蛋白水平。结果与对照组相比,HHT能抑制HT29细胞增殖,诱导细胞核破裂,染色质凝聚,凋亡小体形成;同时,HHT处理能够提高HT29细胞中BAX/Bcl2比值、caspase-3、caspase-9、raptor的mRNA水平,降低PI3K、AKT、rictor的mRNA水平;并上调HT29细胞中c-caspase-3、c-caspase-9、c-PARP、BAX、raptor蛋白水平,下调pro-caspase-3、pro-caspase-9、PARP、PI3K、PDK1、AKT、mTOR、rictor蛋白水平。结论 HHT能够通过阻断mTOR信号通路,进而抑制HT29人结直肠肿瘤细胞增殖并诱导其凋亡。展开更多
目的探讨hsa_circ_0018574在结直肠癌组织和人结肠癌HT29细胞系中的表达,以及其对结直肠肿瘤细胞增殖和凋亡的影响。方法使用circPrimer1.2软件绘制circRNA序列结构示意图,用人circRNA芯片筛选在结直肠癌组织和癌旁组织中差异表达的circ...目的探讨hsa_circ_0018574在结直肠癌组织和人结肠癌HT29细胞系中的表达,以及其对结直肠肿瘤细胞增殖和凋亡的影响。方法使用circPrimer1.2软件绘制circRNA序列结构示意图,用人circRNA芯片筛选在结直肠癌组织和癌旁组织中差异表达的circRNA,提取组织样本RNA,RT-qPCR检测hsa_circ_0018574在人结直肠肿瘤组织中的表达。将si-circ_0018574转染至人结肠癌HT29细胞,分别采用克隆集落形成实验、流式细胞仪、Western blot法检测CDK2、CDK4、CDK6以及cyclinD1和cyclinE周期蛋白表达。结果hsa_circ_0018574在人结直肠肿瘤组织中的表达明显高于癌旁组织(P<0.01)。si-circ_0018574能够明显抑制人结肠癌HT29细胞增殖,减少克隆形成以及降低细胞集落的形成能力(P<0.01),诱导肿瘤细胞凋亡(P<0.01)。CDK2、CDK4、CDK6以及cyclinD1和cyc l inE周期蛋白表达降低。结论hsa_circ_0018574在结直肠肿瘤中高表达,沉默circ_0018574能够显著抑制人结肠癌HT29细胞增殖,减少细胞分裂,诱导细胞凋亡。展开更多
目的观察泛素连接酶肌肉环指状蛋白2(muscle ring finger protein2,MuRF2)在HL-1心肌细胞自噬中的作用。方法采用慢病毒转染技术上调和下调HL-1心肌细胞中MuRF2的表达。实验分为阴性对照组、阳性对照组(空载病毒对照)、雷帕霉素处理组、...目的观察泛素连接酶肌肉环指状蛋白2(muscle ring finger protein2,MuRF2)在HL-1心肌细胞自噬中的作用。方法采用慢病毒转染技术上调和下调HL-1心肌细胞中MuRF2的表达。实验分为阴性对照组、阳性对照组(空载病毒对照)、雷帕霉素处理组、MuRF2过表达组和MuRF2敲减组。用雷帕霉素处理细胞,蛋白免疫印迹法(Western blot)检测自噬蛋白LC3和BNIP3的表达。透射电镜观察HL-1心肌细胞超微结构。结果(1)Western blot验证,MuRF2过表达组中MuRF2蛋白表达水平均高于阴性对照组、阳性对照组、雷帕霉素处理组(P均<0.05),MuRF2敲减组中MuRF2蛋白表达水平均低于阴性对照组、阳性对照组、雷帕霉素处理组(P均<0.05)。(2)Western blot结果表明,雷帕霉素处理组LC3蛋白Ⅱ/Ⅰ比值和BNIP3蛋白表达均高于阴性对照组(P均<0.05),MuRF2过表达组LC3蛋白Ⅱ/Ⅰ比值和BNIP3蛋白表达均低于雷帕霉素处理组(P均<0.05)。与阴性对照组相比,MuRF2敲减组的LC3蛋白Ⅱ/Ⅰ比值和BNIP3蛋白表达均升高(P均<0.05),雷帕霉素处理组和MuRF2敲减组在LC蛋白Ⅱ/Ⅰ比值和BNIP3蛋白表达上差异均无统计学意义(P均>0.05)。透射电镜显示,雷帕霉素处理组线粒体分裂、碎片化,自噬体和自噬溶酶体散在胞质内。MuRF2过表达组线粒体处于融合状态,自噬体明显减少。结论MuRF2可以抑制心肌细胞线粒体自噬。展开更多
目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生...目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生长因子致血管生成的人类脐静脉内皮细胞模型衡量细胞的增殖、黏附及迁移,同时进行细管形成实验及纤维母细胞生长因子致鸡胚绒毛尿囊膜血管生成实验检测WEPPG的抗血管生成作用。结果:WEPPG能够显著抑制纤维母细胞生长因子所致血管生成的人类脐静脉内皮细胞的增殖、黏附及迁移。信号蛋白分析显示多种蛋白表达变化,如细胞周期素A、p63、KIP2的上调及nibrin蛋白和黏着斑激酶的下调。与对照组相比,WEPPG显著减少了鸡胚绒毛尿囊膜血管生成。结论:本研究的结果证实了WEPPG的抗血管生成作用,这可能是这种药物具有抗癌功效的原因之一。展开更多
Olfactory ensheathing cells(OECs) from the olfactory bulb(OB) and the olfactory mucosa(OM) have the capacity to repair nerve injury. However, the difference in the therapeutic effect between OB-derived OECs and OM-der...Olfactory ensheathing cells(OECs) from the olfactory bulb(OB) and the olfactory mucosa(OM) have the capacity to repair nerve injury. However, the difference in the therapeutic effect between OB-derived OECs and OM-derived OECs remains unclear. In this study, we extracted OECs from OB and OM and compared the gene and protein expression profiles of the cells using transcriptomics and non-quantitative proteomics techniques. The results revealed that both OB-derived OECs and OM-derived OECs highly expressed genes and proteins that regulate cell growth, proliferation, apoptosis and vascular endothelial cell regeneration. The differentially expressed genes and proteins of OB-derived OECs play a key role in regulation of nerve regeneration and axon regeneration and extension, transmission of nerve impulses and response to axon injury. The differentially expressed genes and proteins of OM-derived OECs mainly participate in the positive regulation of inflammatory response, defense response, cytokine binding, cell migration and wound healing. These findings suggest that differentially expressed genes and proteins may explain why OB-derived OECs and OM-derived OECs exhibit different therapeutic roles. This study was approved by the Animal Ethics Committee of the General Hospital of Ningxia Medical University(approval No. 2017-073) on February 13, 2017.展开更多
We investigated the anti-angiogenic effects of the water extract of HangAmDan (WEHAD),which is a crude extract of nine Korean medicinal substances of animal and plant origin.In human umbilical vein endothelial cells,W...We investigated the anti-angiogenic effects of the water extract of HangAmDan (WEHAD),which is a crude extract of nine Korean medicinal substances of animal and plant origin.In human umbilical vein endothelial cells,WEHAD significantly inhibited bFGF-induced proliferation,adhesion,migration,and capillary tube formation.We used an antibody array to perform an analysis of signaling proteins,which showed up-regulated expression of various proteins including RAD51,RAD52,and p73,and down-regulated expression of pFAK.Blood vessel formation in a chick chorioallantoic membrane (CAM) treated with WEHAD was markedly reduced in length compared with a PBS-treated control group.These results suggest that inhibition of angiogenesis by WEHAD may be the mechanism of action for the anti-cancer effects of HAD.展开更多
基金Key Research and Development Plan Project of Ningxia Hui Autonomous Region(Project No.:2019BEG03026)Ningxia Overseas Returnees Innovation and Entrepreneurship Project(Project No.:2017-659)+2 种基金Ningxia Key Research and Development Plan Project(Project No.2018BFG02017)Ningxia Medical University Young Backbone Talents Training Program School-level Academic Technology Leader Reserve Cultivation ObjectFirst-Class Discipline Construction Founded Project of Ningxia Medical University and the School of Clinical Medicine(No.NXYLXK2017A05).
文摘Based on butterfly microfluidic gene chip technology,a method for rapid,accurate and efficient detection of 19 common pathogenic microorganisms of nosocomial infection was established,and a butterfly microfluidic gene chip with high-throughput detection was designed and fabricated.Using constant temperature amplification technology,using the polymerase with chain replacement function to react at constant temperature(65℃)and combined with microfluidic chip technology,primers were designed according to the target genes of 19 pathogenic microorganisms,and a butterfly microfluidic gene chip which can detect 19 common pathogenic microorganisms of nosocomial infection was made to simplify the inspection operation process and verify the sensitivity of the chip.The butterfly microfluidic gene chip can be used for the rapid and efficient detection of 19 common pathogenic microorganisms of nosocomial infection,and provides a new idea for the detection and auxiliary diagnosis of pathogenic microorganisms of nosocomial infection.
文摘目的探讨hsa_circ_0018574在结直肠癌组织和人结肠癌HT29细胞系中的表达,以及其对结直肠肿瘤细胞增殖和凋亡的影响。方法使用circPrimer1.2软件绘制circRNA序列结构示意图,用人circRNA芯片筛选在结直肠癌组织和癌旁组织中差异表达的circRNA,提取组织样本RNA,RT-qPCR检测hsa_circ_0018574在人结直肠肿瘤组织中的表达。将si-circ_0018574转染至人结肠癌HT29细胞,分别采用克隆集落形成实验、流式细胞仪、Western blot法检测CDK2、CDK4、CDK6以及cyclinD1和cyclinE周期蛋白表达。结果hsa_circ_0018574在人结直肠肿瘤组织中的表达明显高于癌旁组织(P<0.01)。si-circ_0018574能够明显抑制人结肠癌HT29细胞增殖,减少克隆形成以及降低细胞集落的形成能力(P<0.01),诱导肿瘤细胞凋亡(P<0.01)。CDK2、CDK4、CDK6以及cyclinD1和cyc l inE周期蛋白表达降低。结论hsa_circ_0018574在结直肠肿瘤中高表达,沉默circ_0018574能够显著抑制人结肠癌HT29细胞增殖,减少细胞分裂,诱导细胞凋亡。
文摘目的:本研究旨在证实朝鲜白头翁、人参和甘草的复合水提物(water extract ofPulsatillakoreana(Yabe ex Nakai)Nakai ex T.Mori.,PanaxginsengC.A.Meyer andGlycyrrhizauralensisFisch,WEPPG)的抗血管生成作用。方法:使用纤维母细胞生长因子致血管生成的人类脐静脉内皮细胞模型衡量细胞的增殖、黏附及迁移,同时进行细管形成实验及纤维母细胞生长因子致鸡胚绒毛尿囊膜血管生成实验检测WEPPG的抗血管生成作用。结果:WEPPG能够显著抑制纤维母细胞生长因子所致血管生成的人类脐静脉内皮细胞的增殖、黏附及迁移。信号蛋白分析显示多种蛋白表达变化,如细胞周期素A、p63、KIP2的上调及nibrin蛋白和黏着斑激酶的下调。与对照组相比,WEPPG显著减少了鸡胚绒毛尿囊膜血管生成。结论:本研究的结果证实了WEPPG的抗血管生成作用,这可能是这种药物具有抗癌功效的原因之一。
基金supported by Key Research Projects of Ningxia Hui Autonomous Region of China,No.2018BCG01002(to HCX)the Natural Science Foundation of Ningxia Hui Autonomous Region of China,No.NZ17150(to PY)。
文摘Olfactory ensheathing cells(OECs) from the olfactory bulb(OB) and the olfactory mucosa(OM) have the capacity to repair nerve injury. However, the difference in the therapeutic effect between OB-derived OECs and OM-derived OECs remains unclear. In this study, we extracted OECs from OB and OM and compared the gene and protein expression profiles of the cells using transcriptomics and non-quantitative proteomics techniques. The results revealed that both OB-derived OECs and OM-derived OECs highly expressed genes and proteins that regulate cell growth, proliferation, apoptosis and vascular endothelial cell regeneration. The differentially expressed genes and proteins of OB-derived OECs play a key role in regulation of nerve regeneration and axon regeneration and extension, transmission of nerve impulses and response to axon injury. The differentially expressed genes and proteins of OM-derived OECs mainly participate in the positive regulation of inflammatory response, defense response, cytokine binding, cell migration and wound healing. These findings suggest that differentially expressed genes and proteins may explain why OB-derived OECs and OM-derived OECs exhibit different therapeutic roles. This study was approved by the Animal Ethics Committee of the General Hospital of Ningxia Medical University(approval No. 2017-073) on February 13, 2017.
文摘We investigated the anti-angiogenic effects of the water extract of HangAmDan (WEHAD),which is a crude extract of nine Korean medicinal substances of animal and plant origin.In human umbilical vein endothelial cells,WEHAD significantly inhibited bFGF-induced proliferation,adhesion,migration,and capillary tube formation.We used an antibody array to perform an analysis of signaling proteins,which showed up-regulated expression of various proteins including RAD51,RAD52,and p73,and down-regulated expression of pFAK.Blood vessel formation in a chick chorioallantoic membrane (CAM) treated with WEHAD was markedly reduced in length compared with a PBS-treated control group.These results suggest that inhibition of angiogenesis by WEHAD may be the mechanism of action for the anti-cancer effects of HAD.
