The aim of this study was to investigate the knockdown efficiency of 2＇-O-methylated （2＇-OMe）-modified small interfering RNAs （siRNAs） on human rhinovirus 1B （HRV1B） replication and the interferon response. Th...The aim of this study was to investigate the knockdown efficiency of 2＇-O-methylated （2＇-OMe）-modified small interfering RNAs （siRNAs） on human rhinovirus 1B （HRV1B） replication and the interferon response. Thus, 24 2＇-OMe-modified siRNAs were designed to target HRV1B. The RNA levels of HRV1B, Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and interferons were determined in HRV1B-infected HeLa and BEAS-2B epithelial cells transfected with 2＇-OMe-modified siRNAs. The results revealed that all 2＇-OMe-modified siRNAs interfered with the replication of HRVIB in a cell-specific and transfection efficiency-dependent manner. Viral activation of Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-1, and the interferon response was detected. In conclusion, the 2＇-OMe-modified siRNAs used in this study could interfere with HRV1B replication, possibly leading to the reactivation of the interferon response.展开更多
目的研究中药复方桃红四物汤(Tao Hong Si Wu decoction,THSWD)治疗大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠长链非编码RNA(long non-coding RNA,lncRNA)的表达,并确定THSWD治疗MCAO大鼠可能的分子机制。方法从对照...目的研究中药复方桃红四物汤(Tao Hong Si Wu decoction,THSWD)治疗大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠长链非编码RNA(long non-coding RNA,lncRNA)的表达,并确定THSWD治疗MCAO大鼠可能的分子机制。方法从对照组、MCAO组和MCAO+THSWD组各获得3个大脑半球组织。采用RNA测序技术鉴定三组中的lncRNA基因表达。鉴定了THSWD调节的lncRNA基因,然后构建了THSWD调节的lncRNA-mRNA网络。通过MCODE插件鉴定lncRNA-mRNA网络的模块。基因本体(gene ontology,GO)和京都基因与基因组百科全书数据库(kyoto encyclopedia of genes and genomes,KEGG)用于分析富集的生物功能和信号通路。鉴定了THSWD调节的lncRNA的顺式和反式调控基因。采用逆转录实时定量聚合酶链式反应(RT-qPCR)验证lncRNA。分子对接用于验证lncRNA-mRNA网络靶点和通路相关蛋白结合能力。结果在MCAO大鼠中,THSWD共调节了302个lncRNA。生物信息学分析表明,一些核心lncRNA可能在THSWD治疗MCAO大鼠中发挥重要作用,此外,我们进一步发现THSWD可能也通过lncRNA-mRNA网络以及网络富集的补体和凝血级联反应等多通路治疗MCAO大鼠。分子对接结果表明,THSWD活性化合物没食子酸和苦杏仁苷与蛋白质靶点具有一定的结合能力。结论THSWD可以通过调节lncRNA保护MCAO大鼠脑损伤,为THSWD治疗缺血性中风提供了新见解。展开更多
The full-length c DNA of functionally-unknown salivary protein C002 in Schizaphis graminum was cloned using rapid amplification of c DNA ends（RACE） and designated as Sg C002（Gen Bank accession no. KC977563）. It is...The full-length c DNA of functionally-unknown salivary protein C002 in Schizaphis graminum was cloned using rapid amplification of c DNA ends（RACE） and designated as Sg C002（Gen Bank accession no. KC977563）. It is 767 bp long and encodes a protein of 190 amino acid residues with a predicted mass of 21.5 k Da and a predicted cleavage site of N-terminal signal peptide between the 24 th and the 25 th residues. Sg C002 is specifically expressed in salivary gland with the highest level at the 2nd instar. Introducing Sg C002-specific 476-si RNA, but not 546-si RNA to aphids through artificial diet significantly suppressed Sg C002 expression. Silencing Sg C002 gene led to lethality of the aphid on wheat plants, but not on pure artificial diet. Our study demonstrated that artificial diet-mediated RNAi can be a useful tool for research on the roles of genes in aphid salivary gland, and also provided new insights into the characteristics of C002 in wheat aphids.展开更多
目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)...目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)组(加K-ras si RNA和Sirtinol处理);Western blot检测SIRT1蛋白的表达情况,Q-PCR检测K-ras m RNA水平和周期蛋白Cyclin D1 m RNA水平,MTT检测细胞的增殖活力,流式细胞仪检测细胞的凋亡情况.结果:Western blot结果显示相对于C组和K组,S组和(K+S)组中的SIRT1蛋白的表达明显下降;Q-PCR结果显示K组和(K+S)组中的K-ras m RNA水平分别是C组的0.454±0.037、0.413±0.032倍,差异具有统计意义;MTT结果显示C组、K组、S组和(K+S)组的A值分别是0.814±0.025、0.634±0.038、0.613±0.036、0.401±0.019,相对于C组,K组、S组和(K+S)组的A值明显下降,其中(K+S)组最明显;Q-PCR结果显示K组、S组和(K+S)组中的Cyclin D1 m RNA水平分别是C组的0.693倍±0.046倍、0.634倍±0.032倍、0.400倍±0.034倍,差异具有统计学意义,其中(K+S)组下降最明显;流式细胞仪显示C组、K组、S组和(K+S)组的凋亡率分别是4.29%±0.246%、7.4 6 9%±0.4 5 7%、8.2 0 6%±0.4 9 0%、12.272%±0.675%,相对于C组,K组、S组和(K+S)组的凋亡率明显上升,其中(K+S)组最明显.结论:Sirtinol联合K-ras si RNA可以更明显的抑制胰腺癌PANC-1细胞的增殖和促进其凋亡.展开更多
The lentivirus-mediated u PA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white r...The lentivirus-mediated u PA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white rabbits. They were cultured with interleukin（IL）-1β（10 ng/m L） for 24 h and then divided into three groups： u PA-si RNA group（cells transfected with u PA-si RNA lentiviruses）, blank control group（untreated cells）, and negative control group（cells transfected with empty vectors）. Western blotting and real-time quantitative reverse transcription-PCR（RT-QPCR） were performed to detect the protein and m RNA expression levels of u PA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13 and MMP-14 in osteoarthritic chondrocytes. Cell Counting Kit-8, flow cytometry, and colony formation assay were used to examine the proliferation and apoptosis of chondrocytes. The results showed that after u PA-si RNA transfection, the protein and mR NA expression levels of uP A, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13, and MMP-14 were significantly decreased（P〈0.05 for MMP-1, MMP-9, MMP-10 and MMP-14, P〈0.01 for u PA, MMP-3 and MMP-13）. Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in u PA-si RNA group than in control groups（P〈0.01）. The proportion of cells in G0/G1 phase was markedly increased and that in the S phase decreased, and the cell cycle was arrested at the G1/S phase in the control group. In the u PAsi RNA group, the proportion of cells in the S phase was significantly increased, resulting in a different proportion of cells in cell cycle phase（P〈0.01）. It was suggested that the down-regulation of uP A gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.展开更多
文摘The aim of this study was to investigate the knockdown efficiency of 2＇-O-methylated （2＇-OMe）-modified small interfering RNAs （siRNAs） on human rhinovirus 1B （HRV1B） replication and the interferon response. Thus, 24 2＇-OMe-modified siRNAs were designed to target HRV1B. The RNA levels of HRV1B, Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-I, and interferons were determined in HRV1B-infected HeLa and BEAS-2B epithelial cells transfected with 2＇-OMe-modified siRNAs. The results revealed that all 2＇-OMe-modified siRNAs interfered with the replication of HRVIB in a cell-specific and transfection efficiency-dependent manner. Viral activation of Toll-like receptor 3, melanoma differentiation-associated gene 5, retinoic acid inducible gene-1, and the interferon response was detected. In conclusion, the 2＇-OMe-modified siRNAs used in this study could interfere with HRV1B replication, possibly leading to the reactivation of the interferon response.
文摘目的研究中药复方桃红四物汤(Tao Hong Si Wu decoction,THSWD)治疗大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)大鼠长链非编码RNA(long non-coding RNA,lncRNA)的表达,并确定THSWD治疗MCAO大鼠可能的分子机制。方法从对照组、MCAO组和MCAO+THSWD组各获得3个大脑半球组织。采用RNA测序技术鉴定三组中的lncRNA基因表达。鉴定了THSWD调节的lncRNA基因,然后构建了THSWD调节的lncRNA-mRNA网络。通过MCODE插件鉴定lncRNA-mRNA网络的模块。基因本体(gene ontology,GO)和京都基因与基因组百科全书数据库(kyoto encyclopedia of genes and genomes,KEGG)用于分析富集的生物功能和信号通路。鉴定了THSWD调节的lncRNA的顺式和反式调控基因。采用逆转录实时定量聚合酶链式反应(RT-qPCR)验证lncRNA。分子对接用于验证lncRNA-mRNA网络靶点和通路相关蛋白结合能力。结果在MCAO大鼠中,THSWD共调节了302个lncRNA。生物信息学分析表明,一些核心lncRNA可能在THSWD治疗MCAO大鼠中发挥重要作用,此外,我们进一步发现THSWD可能也通过lncRNA-mRNA网络以及网络富集的补体和凝血级联反应等多通路治疗MCAO大鼠。分子对接结果表明,THSWD活性化合物没食子酸和苦杏仁苷与蛋白质靶点具有一定的结合能力。结论THSWD可以通过调节lncRNA保护MCAO大鼠脑损伤,为THSWD治疗缺血性中风提供了新见解。
基金supported by the National Natural Science Foundation of China (30971920,31371946)the International Cooperation Project between China and Belgium (2010DFA32810)the Earmarked Fund for Modern Agro-Industry Technology Research System,China (CARS3)
文摘The full-length c DNA of functionally-unknown salivary protein C002 in Schizaphis graminum was cloned using rapid amplification of c DNA ends（RACE） and designated as Sg C002（Gen Bank accession no. KC977563）. It is 767 bp long and encodes a protein of 190 amino acid residues with a predicted mass of 21.5 k Da and a predicted cleavage site of N-terminal signal peptide between the 24 th and the 25 th residues. Sg C002 is specifically expressed in salivary gland with the highest level at the 2nd instar. Introducing Sg C002-specific 476-si RNA, but not 546-si RNA to aphids through artificial diet significantly suppressed Sg C002 expression. Silencing Sg C002 gene led to lethality of the aphid on wheat plants, but not on pure artificial diet. Our study demonstrated that artificial diet-mediated RNAi can be a useful tool for research on the roles of genes in aphid salivary gland, and also provided new insights into the characteristics of C002 in wheat aphids.
文摘目的:探讨Sirtinol联合K-ras基因RNA干扰对胰腺癌PANC-1细胞增殖和凋亡的影响.方法:用50μmol/m L的Sirtinol和50 nmol K-ras si RNA处理胰腺癌PANC-1细胞48 h,分为C组(无加药处理),K组(加K-ras si RNA处理),S组(加Sirtinol处理),(K+S)组(加K-ras si RNA和Sirtinol处理);Western blot检测SIRT1蛋白的表达情况,Q-PCR检测K-ras m RNA水平和周期蛋白Cyclin D1 m RNA水平,MTT检测细胞的增殖活力,流式细胞仪检测细胞的凋亡情况.结果:Western blot结果显示相对于C组和K组,S组和(K+S)组中的SIRT1蛋白的表达明显下降;Q-PCR结果显示K组和(K+S)组中的K-ras m RNA水平分别是C组的0.454±0.037、0.413±0.032倍,差异具有统计意义;MTT结果显示C组、K组、S组和(K+S)组的A值分别是0.814±0.025、0.634±0.038、0.613±0.036、0.401±0.019,相对于C组,K组、S组和(K+S)组的A值明显下降,其中(K+S)组最明显;Q-PCR结果显示K组、S组和(K+S)组中的Cyclin D1 m RNA水平分别是C组的0.693倍±0.046倍、0.634倍±0.032倍、0.400倍±0.034倍,差异具有统计学意义,其中(K+S)组下降最明显;流式细胞仪显示C组、K组、S组和(K+S)组的凋亡率分别是4.29%±0.246%、7.4 6 9%±0.4 5 7%、8.2 0 6%±0.4 9 0%、12.272%±0.675%,相对于C组,K组、S组和(K+S)组的凋亡率明显上升,其中(K+S)组最明显.结论:Sirtinol联合K-ras si RNA可以更明显的抑制胰腺癌PANC-1细胞的增殖和促进其凋亡.
基金supported by grants from the National Natural Science Foundation of China(Nos.81160225,812604531,and 81360451)the Xinjiang Bingtuan Special Program of Medical Science(Nos.2011BC004,2013BA020,and 2014BC003)
文摘The lentivirus-mediated u PA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white rabbits. They were cultured with interleukin（IL）-1β（10 ng/m L） for 24 h and then divided into three groups： u PA-si RNA group（cells transfected with u PA-si RNA lentiviruses）, blank control group（untreated cells）, and negative control group（cells transfected with empty vectors）. Western blotting and real-time quantitative reverse transcription-PCR（RT-QPCR） were performed to detect the protein and m RNA expression levels of u PA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13 and MMP-14 in osteoarthritic chondrocytes. Cell Counting Kit-8, flow cytometry, and colony formation assay were used to examine the proliferation and apoptosis of chondrocytes. The results showed that after u PA-si RNA transfection, the protein and mR NA expression levels of uP A, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13, and MMP-14 were significantly decreased（P〈0.05 for MMP-1, MMP-9, MMP-10 and MMP-14, P〈0.01 for u PA, MMP-3 and MMP-13）. Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in u PA-si RNA group than in control groups（P〈0.01）. The proportion of cells in G0/G1 phase was markedly increased and that in the S phase decreased, and the cell cycle was arrested at the G1/S phase in the control group. In the u PAsi RNA group, the proportion of cells in the S phase was significantly increased, resulting in a different proportion of cells in cell cycle phase（P〈0.01）. It was suggested that the down-regulation of uP A gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.
