CORD-19:0d5be5a4268da7c08b478ca36721d46bd67bbe69 JSONTXT 8 Projects

Annnotations TAB TSV DIC JSON TextAE-old TextAE

Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-114 Sentence denotes Effect of avian influenza A H5N1 infection on the expression of microRNA-141 in human respiratory epithelial cells
TextSentencer_T1 0-114 Sentence denotes Effect of avian influenza A H5N1 infection on the expression of microRNA-141 in human respiratory epithelial cells
TextSentencer_T2 116-124 Sentence denotes Abstract
TextSentencer_T2 116-124 Sentence denotes Abstract
TextSentencer_T3 125-136 Sentence denotes Background:
TextSentencer_T3 125-136 Sentence denotes Background:
TextSentencer_T4 137-194 Sentence denotes Avian influenza remains a serious threat to human health.
TextSentencer_T4 137-194 Sentence denotes Avian influenza remains a serious threat to human health.
TextSentencer_T5 195-298 Sentence denotes The consequence of human infection varies markedly among different subtypes of avian influenza viruses.
TextSentencer_T5 195-298 Sentence denotes The consequence of human infection varies markedly among different subtypes of avian influenza viruses.
TextSentencer_T6 299-404 Sentence denotes In addition to viral factors, the difference in host cellular response is likely to play a critical role.
TextSentencer_T6 299-404 Sentence denotes In addition to viral factors, the difference in host cellular response is likely to play a critical role.
TextSentencer_T7 405-618 Sentence denotes This study aims at elucidating how avian influenza infection perturbs the host's miRNA regulatory pathways that may lead to adverse pathological events, such as cytokine storm, using the miRNA microarray approach.
TextSentencer_T7 405-618 Sentence denotes This study aims at elucidating how avian influenza infection perturbs the host's miRNA regulatory pathways that may lead to adverse pathological events, such as cytokine storm, using the miRNA microarray approach.
TextSentencer_T8 619-627 Sentence denotes Results:
TextSentencer_T8 619-627 Sentence denotes Results:
TextSentencer_T9 628-760 Sentence denotes The results showed that dysregulation of miRNA expression was mainly observed in highly pathogenic avian influenza A H5N1 infection.
TextSentencer_T9 628-760 Sentence denotes The results showed that dysregulation of miRNA expression was mainly observed in highly pathogenic avian influenza A H5N1 infection.
TextSentencer_T10 761-911 Sentence denotes We found that miR-21*, miR-100*, miR-141, miR-574-3p, miR-1274a and miR1274b were differentially expressed in response to influenza A virus infection.
TextSentencer_T10 761-911 Sentence denotes We found that miR-21*, miR-100*, miR-141, miR-574-3p, miR-1274a and miR1274b were differentially expressed in response to influenza A virus infection.
TextSentencer_T11 912-1113 Sentence denotes Interestingly, we demonstrated that miR-141, which was more highly induced by H5N1 than by H1N1 (p < 0.05), had an ability to suppress the expression of a cytokine -transforming growth factor (TGF)-β2.
TextSentencer_T11 912-1113 Sentence denotes Interestingly, we demonstrated that miR-141, which was more highly induced by H5N1 than by H1N1 (p < 0.05), had an ability to suppress the expression of a cytokine -transforming growth factor (TGF)-β2.
TextSentencer_T12 1114-1214 Sentence denotes This was supported by the observation that the inhibitory effect could be reversed by antagomiR-141.
TextSentencer_T12 1114-1214 Sentence denotes This was supported by the observation that the inhibitory effect could be reversed by antagomiR-141.
TextSentencer_T13 1215-1227 Sentence denotes Conclusions:
TextSentencer_T13 1215-1227 Sentence denotes Conclusions:
TextSentencer_T14 1228-1780 Sentence denotes Since TGF-β2 is an important cytokine that can act as both an immunosuppressive agent and a potent proinflammatory molecule through its ability to attract and regulate inflammatory molecules, and previous report showed that only seasonal influenza H1N1 (but not the other avian influenza subtypes) could induce a persistent expression of TGF-β2, we speculate that the modulation of TGF-β2 expression by different influenza subtypes via miR-141 might be a critical step for determining the outcome of either normal or excessive inflammation progression.
TextSentencer_T14 1228-1780 Sentence denotes Since TGF-β2 is an important cytokine that can act as both an immunosuppressive agent and a potent proinflammatory molecule through its ability to attract and regulate inflammatory molecules, and previous report showed that only seasonal influenza H1N1 (but not the other avian influenza subtypes) could induce a persistent expression of TGF-β2, we speculate that the modulation of TGF-β2 expression by different influenza subtypes via miR-141 might be a critical step for determining the outcome of either normal or excessive inflammation progression.
TextSentencer_T15 1782-1851 Sentence denotes Avian influenza remains a serious threat to poultry and human health.
TextSentencer_T15 1782-1851 Sentence denotes Avian influenza remains a serious threat to poultry and human health.
TextSentencer_T16 1852-1989 Sentence denotes From December 2003 to April 2013, more than 600 human infections and 374 deaths have been reported to the World Health Organization [1] .
TextSentencer_T16 1852-1989 Sentence denotes From December 2003 to April 2013, more than 600 human infections and 374 deaths have been reported to the World Health Organization [1] .
TextSentencer_T17 1990-2072 Sentence denotes Outbreaks of H5N1 in poultry swept from Southeast Asia to many parts of the world.
TextSentencer_T17 1990-2072 Sentence denotes Outbreaks of H5N1 in poultry swept from Southeast Asia to many parts of the world.
TextSentencer_T18 2073-2140 Sentence denotes To date, there is still no sign that the epidemic is under control.
TextSentencer_T18 2073-2140 Sentence denotes To date, there is still no sign that the epidemic is under control.
TextSentencer_T19 2141-2318 Sentence denotes While it has been well documented that infection with H5N1 results in high mortality in humans [2] [3] [4] [5] , the cellular pathway leading to such adverse outcome is unknown.
TextSentencer_T19 2141-2318 Sentence denotes While it has been well documented that infection with H5N1 results in high mortality in humans [2] [3] [4] [5] , the cellular pathway leading to such adverse outcome is unknown.
TextSentencer_T20 2319-2474 Sentence denotes The naive host immune system cannot be the sole explanation as infection of other avian influenza viruses, e.g. H9N2, only results in mild infections [6] .
TextSentencer_T20 2319-2474 Sentence denotes The naive host immune system cannot be the sole explanation as infection of other avian influenza viruses, e.g. H9N2, only results in mild infections [6] .
TextSentencer_T21 2475-2852 Sentence denotes While the predilection of H5N1 towards cells in the lower respiratory tract contributes to the development of severe pneumonia [7] , the available clinico-pathological evidence indicates that the infected patients progress to multi-organ failure early in the course of illness, and the degree of organ failure is out of proportion to the involvement of infection [8] [9] [10] .
TextSentencer_T21 2475-2852 Sentence denotes While the predilection of H5N1 towards cells in the lower respiratory tract contributes to the development of severe pneumonia [7] , the available clinico-pathological evidence indicates that the infected patients progress to multi-organ failure early in the course of illness, and the degree of organ failure is out of proportion to the involvement of infection [8] [9] [10] .
TextSentencer_T22 2853-2990 Sentence denotes Cytokine storm and reactive haemophagocytic syndrome are the key features that distinguish H5N1 infection from severe seasonal influenza.
TextSentencer_T22 2853-2990 Sentence denotes Cytokine storm and reactive haemophagocytic syndrome are the key features that distinguish H5N1 infection from severe seasonal influenza.
TextSentencer_T23 2991-3112 Sentence denotes These indirect mechanisms seem to play an even more important role than direct cell killing due to lytic viral infection.
TextSentencer_T23 2991-3112 Sentence denotes These indirect mechanisms seem to play an even more important role than direct cell killing due to lytic viral infection.
TextSentencer_T24 3113-3258 Sentence denotes MiRNAs, a new class of endogenous, 18-23 nucleotide long noncoding and single-stranded RNAs, were recently discovered in both animals and plants.
TextSentencer_T24 3113-3258 Sentence denotes MiRNAs, a new class of endogenous, 18-23 nucleotide long noncoding and single-stranded RNAs, were recently discovered in both animals and plants.
TextSentencer_T25 3259-3387 Sentence denotes They trigger translational repression and/or mRNA degradation mostly through complementary binding to the 3′UTR of target mRNAs.
TextSentencer_T25 3259-3387 Sentence denotes They trigger translational repression and/or mRNA degradation mostly through complementary binding to the 3′UTR of target mRNAs.
TextSentencer_T26 3388-3549 Sentence denotes Studies have shown that miRNAs can regulate a wide array of biological processes such as cell proliferation, differentiation, and apoptosis [11] [12] [13] [14] .
TextSentencer_T26 3388-3549 Sentence denotes Studies have shown that miRNAs can regulate a wide array of biological processes such as cell proliferation, differentiation, and apoptosis [11] [12] [13] [14] .
TextSentencer_T27 3550-3812 Sentence denotes Given the nature of viruses, being intracellular parasites and using the cellular machinery for their survival and replication, the success of the virus essentially depends on its ability to effectively and efficiently use the host machinery to propagate itself.
TextSentencer_T27 3550-3812 Sentence denotes Given the nature of viruses, being intracellular parasites and using the cellular machinery for their survival and replication, the success of the virus essentially depends on its ability to effectively and efficiently use the host machinery to propagate itself.
TextSentencer_T28 3813-4003 Sentence denotes This dependence on the host also makes it susceptible to the host gene-regulatory mechanisms, i.e. the host miRNAs may also have direct or indirect regulatory role on viral mRNAs expression.
TextSentencer_T28 3813-4003 Sentence denotes This dependence on the host also makes it susceptible to the host gene-regulatory mechanisms, i.e. the host miRNAs may also have direct or indirect regulatory role on viral mRNAs expression.
TextSentencer_T29 4004-4122 Sentence denotes Recently, several reports indicated that miRNAs can target influenza viruses and regulate influenza virus replication.
TextSentencer_T29 4004-4122 Sentence denotes Recently, several reports indicated that miRNAs can target influenza viruses and regulate influenza virus replication.
TextSentencer_T30 4123-4320 Sentence denotes In one report, 36 pig-encoded miRNAs and 22 human-encoded miRNAs were found to have putative targets in swine influenza virus and Swine-Origin 2009 A/H1N1 influenza virus genes, respectively [15] .
TextSentencer_T30 4123-4320 Sentence denotes In one report, 36 pig-encoded miRNAs and 22 human-encoded miRNAs were found to have putative targets in swine influenza virus and Swine-Origin 2009 A/H1N1 influenza virus genes, respectively [15] .
TextSentencer_T31 4321-4503 Sentence denotes In another report, results showed that miR-323, miR-491 and miR-654 could inhibit replication of H1N1 influenza A virus through binding to the conserved region of the PB1 gene [16] .
TextSentencer_T31 4321-4503 Sentence denotes In another report, results showed that miR-323, miR-491 and miR-654 could inhibit replication of H1N1 influenza A virus through binding to the conserved region of the PB1 gene [16] .
TextSentencer_T32 4504-4616 Sentence denotes These miRNAs could downregulate PB1 expression through mRNA degradation instead of translation repression [16] .
TextSentencer_T32 4504-4616 Sentence denotes These miRNAs could downregulate PB1 expression through mRNA degradation instead of translation repression [16] .
TextSentencer_T33 4617-4788 Sentence denotes Besides targeting influenza virus, cellular miRNAs were also implicated in the lethal infections of mice with a highly pathogenic 1918 pandemic H1N1 influenza virus [17] .
TextSentencer_T33 4617-4788 Sentence denotes Besides targeting influenza virus, cellular miRNAs were also implicated in the lethal infections of mice with a highly pathogenic 1918 pandemic H1N1 influenza virus [17] .
TextSentencer_T34 4789-5056 Sentence denotes A previous study on miRNA gene expression in avian influenza virus infected chicken showed that miR-146, which was previously reported to be associated with immune-related signal pathways in mammals, was found to be differentially expressed in infected tissues [18] .
TextSentencer_T34 4789-5056 Sentence denotes A previous study on miRNA gene expression in avian influenza virus infected chicken showed that miR-146, which was previously reported to be associated with immune-related signal pathways in mammals, was found to be differentially expressed in infected tissues [18] .
TextSentencer_T35 5057-5352 Sentence denotes Moreover, a study of profiling cellular miRNAs of lung tissue from cynomolgus macaques infected with a highly pathogenic H5N1 avian and a less pathogenic 1918 H1N1 reassortant virus identified that 23 miRNAs were associated with the extreme virulence of highly pathogenic H5N1 avian virus [19] .
TextSentencer_T35 5057-5352 Sentence denotes Moreover, a study of profiling cellular miRNAs of lung tissue from cynomolgus macaques infected with a highly pathogenic H5N1 avian and a less pathogenic 1918 H1N1 reassortant virus identified that 23 miRNAs were associated with the extreme virulence of highly pathogenic H5N1 avian virus [19] .
TextSentencer_T36 5353-5524 Sentence denotes Also, the predicted gene targets of the identified miRNAs were found to be associated with aberrant and uncontrolled inflammatory responses and increased cell death [19] .
TextSentencer_T36 5353-5524 Sentence denotes Also, the predicted gene targets of the identified miRNAs were found to be associated with aberrant and uncontrolled inflammatory responses and increased cell death [19] .
TextSentencer_T37 5525-5691 Sentence denotes This study aimed at elucidating how avian influenza infection perturbs the human gene regulatory pathways leading to adverse pathological events, e.g. cytokine storm.
TextSentencer_T37 5525-5691 Sentence denotes This study aimed at elucidating how avian influenza infection perturbs the human gene regulatory pathways leading to adverse pathological events, e.g. cytokine storm.
TextSentencer_T38 5692-5848 Sentence denotes We hypothesized that miRNAs could be involved in influenza virus infection response and began addressing this hypothesis using a microarray-based screening.
TextSentencer_T38 5692-5848 Sentence denotes We hypothesized that miRNAs could be involved in influenza virus infection response and began addressing this hypothesis using a microarray-based screening.
TextSentencer_T39 5849-6024 Sentence denotes The ultimate goal of this study is to generate essential information for further studies to identify novel intervention targets to ameliorate the adverse outcome of infection.
TextSentencer_T39 5849-6024 Sentence denotes The ultimate goal of this study is to generate essential information for further studies to identify novel intervention targets to ameliorate the adverse outcome of infection.
TextSentencer_T40 6025-6162 Sentence denotes The cell line -NCI-H292, infected with various preparations of influenza viruses was analysed for miRNA expression profiles subsequently.
TextSentencer_T40 6025-6162 Sentence denotes The cell line -NCI-H292, infected with various preparations of influenza viruses was analysed for miRNA expression profiles subsequently.
TextSentencer_T41 6163-6326 Sentence denotes A list of differentially expressed miRNA was identified for subtypes H1N1 and H5N1, respectively (Table 1) , and the temporal pattern of expression was delineated.
TextSentencer_T41 6163-6326 Sentence denotes A list of differentially expressed miRNA was identified for subtypes H1N1 and H5N1, respectively (Table 1) , and the temporal pattern of expression was delineated.
TextSentencer_T42 6327-6592 Sentence denotes Among the listed profiles of differentially up-regulated miRNA, it was found that miR-141, miR-181c*, miR-210, miR29b, miR-324-5p, and miR-663 were up-regulated (>1.5-fold, p<0.05) at 3-hour post-infection with subtype H5 as compared with noninfected control cells.
TextSentencer_T42 6327-6592 Sentence denotes Among the listed profiles of differentially up-regulated miRNA, it was found that miR-141, miR-181c*, miR-210, miR29b, miR-324-5p, and miR-663 were up-regulated (>1.5-fold, p<0.05) at 3-hour post-infection with subtype H5 as compared with noninfected control cells.
TextSentencer_T43 6593-6688 Sentence denotes At this time point, only miR-141 was found to be slightly induced in subtype H1 infected cells.
TextSentencer_T43 6593-6688 Sentence denotes At this time point, only miR-141 was found to be slightly induced in subtype H1 infected cells.
TextSentencer_T44 6689-6888 Sentence denotes At 6-hour post-infection, it was found that miR-483-3p was up-regulated (>3-fold, p<0.05) in H5N1 infected cells while miR-663 was found to be up-regulated (>1.5-fold, p<0.05) in H1N1 infected cells.
TextSentencer_T44 6689-6888 Sentence denotes At 6-hour post-infection, it was found that miR-483-3p was up-regulated (>3-fold, p<0.05) in H5N1 infected cells while miR-663 was found to be up-regulated (>1.5-fold, p<0.05) in H1N1 infected cells.
TextSentencer_T45 6889-7024 Sentence denotes At 18 and 24-hour postinfection, miR-923, miR-1246, miR-574-3p, and miR-663 were up-regulated (>3-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T45 6889-7024 Sentence denotes At 18 and 24-hour postinfection, miR-923, miR-1246, miR-574-3p, and miR-663 were up-regulated (>3-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T46 7025-7273 Sentence denotes For H1N1 infected cells, at 18 and 24-hour post-infection, miR-188-5p, miR-1260, miR-1274a, miR-1274b, miR141, miR183*, miR-18b, miR-19a, miR21*, miR-301a, miR-572, miR-720, and miR-939 were found to be up-regulated (>1.5-fold, p<0.05) ( Table 1) .
TextSentencer_T46 7025-7273 Sentence denotes For H1N1 infected cells, at 18 and 24-hour post-infection, miR-188-5p, miR-1260, miR-1274a, miR-1274b, miR141, miR183*, miR-18b, miR-19a, miR21*, miR-301a, miR-572, miR-720, and miR-939 were found to be up-regulated (>1.5-fold, p<0.05) ( Table 1) .
TextSentencer_T47 7274-7496 Sentence denotes Among the listed profiles of differentially downregulated miRNA as compared with non-infected control cells, it was found that miR-574-5p was down regulated (>2-fold, p<0.05) in H5N1 infected cells at 3-hour postinfection.
TextSentencer_T47 7274-7496 Sentence denotes Among the listed profiles of differentially downregulated miRNA as compared with non-infected control cells, it was found that miR-574-5p was down regulated (>2-fold, p<0.05) in H5N1 infected cells at 3-hour postinfection.
TextSentencer_T48 7497-7615 Sentence denotes For H1N1 infected cells, miR-23a, miR-574-3p and miR-574-5p were down-regulated (>=2-fold, p<0.05) at this time point.
TextSentencer_T48 7497-7615 Sentence denotes For H1N1 infected cells, miR-23a, miR-574-3p and miR-574-5p were down-regulated (>=2-fold, p<0.05) at this time point.
TextSentencer_T49 7616-7778 Sentence denotes At 6-hour post-infection, miR-126, miR-20a*, miR-362-5p, miR-378, miR-454, and miR574-5p were found to be down-regulated (>2-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T49 7616-7778 Sentence denotes At 6-hour post-infection, miR-126, miR-20a*, miR-362-5p, miR-378, miR-454, and miR574-5p were found to be down-regulated (>2-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T50 7779-7950 Sentence denotes At the same time point (6-hour), miR-15a*, miR-1825, miR-183*, miR-34b, miR-494, and miR-574-5p were found to be down-regulated (>1.5-fold, p<0.05) in H1N1 infected cells.
TextSentencer_T50 7779-7950 Sentence denotes At the same time point (6-hour), miR-15a*, miR-1825, miR-183*, miR-34b, miR-494, and miR-574-5p were found to be down-regulated (>1.5-fold, p<0.05) in H1N1 infected cells.
TextSentencer_T51 7951-8179 Sentence denotes Furthermore, at 18, and 24-hour post-infection, miR-1260, miR-1274a, miR-1274b, miR-141, miR-18b, miR-21*, miR-720, miR-100*, miR-1260, miR1280, and miR21* were found to be downregulated (>3-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T51 7951-8179 Sentence denotes Furthermore, at 18, and 24-hour post-infection, miR-1260, miR-1274a, miR-1274b, miR-141, miR-18b, miR-21*, miR-720, miR-100*, miR-1260, miR1280, and miR21* were found to be downregulated (>3-fold, p<0.05) in H5N1 infected cells.
TextSentencer_T52 8180-8315 Sentence denotes At these time points, only miR-1274, and miR-17* were found to be down-regulated (>1.8-fold, p<0.05) in H1N1 infected cells (Table 1) .
TextSentencer_T52 8180-8315 Sentence denotes At these time points, only miR-1274, and miR-17* were found to be down-regulated (>1.8-fold, p<0.05) in H1N1 infected cells (Table 1) .
TextSentencer_T53 8316-8428 Sentence denotes From the results, we found that similar changes in miRNA profiles were observed in both H1N1 and H5N1 infection.
TextSentencer_T53 8316-8428 Sentence denotes From the results, we found that similar changes in miRNA profiles were observed in both H1N1 and H5N1 infection.
TextSentencer_T54 8429-8545 Sentence denotes However, the magnitude of fold-changes which occurred in H1N1 infection were much lower than that in H5N1 infection.
TextSentencer_T54 8429-8545 Sentence denotes However, the magnitude of fold-changes which occurred in H1N1 infection were much lower than that in H5N1 infection.
TextSentencer_T55 8546-8639 Sentence denotes The microarray data were further confirmed using TaqMan quantitative RT-PCR (qRT-PCR) assays.
TextSentencer_T55 8546-8639 Sentence denotes The microarray data were further confirmed using TaqMan quantitative RT-PCR (qRT-PCR) assays.
TextSentencer_T56 8640-8724 Sentence denotes There were general consistency between TaqMan qRT-PCR assays and microarray results.
TextSentencer_T56 8640-8724 Sentence denotes There were general consistency between TaqMan qRT-PCR assays and microarray results.
TextSentencer_T57 8725-8874 Sentence denotes It was found that six miRNAs (miR-21*, miR-100*, miR-141, miR-1274a, miR-1274b and miR-574-3p) were initially up-regulated at 3 hours post-infection.
TextSentencer_T57 8725-8874 Sentence denotes It was found that six miRNAs (miR-21*, miR-100*, miR-141, miR-1274a, miR-1274b and miR-574-3p) were initially up-regulated at 3 hours post-infection.
TextSentencer_T58 8875-9016 Sentence denotes The degree of up-regulation was more prominent in H5N1 infection (5 to 14 folds) (p*<0.05) than in H1N1 infection (1.5 to 3 folds) (p*<0.05).
TextSentencer_T58 8875-9016 Sentence denotes The degree of up-regulation was more prominent in H5N1 infection (5 to 14 folds) (p*<0.05) than in H1N1 infection (1.5 to 3 folds) (p*<0.05).
TextSentencer_T59 9017-9111 Sentence denotes It was also found that these miRNAs became down-regulated during 6-to-24 hours post-infection.
TextSentencer_T59 9017-9111 Sentence denotes It was also found that these miRNAs became down-regulated during 6-to-24 hours post-infection.
TextSentencer_T60 9112-9212 Sentence denotes The degree of down-regulation was also higher in H5N1 infection than in H1N1 infection ( Figure 1 ).
TextSentencer_T60 9112-9212 Sentence denotes The degree of down-regulation was also higher in H5N1 infection than in H1N1 infection ( Figure 1 ).
TextSentencer_T61 9213-9466 Sentence denotes We then examined the list of targets predicted by TargetScan computer software (http://www.targetscan. org/) for the miRNA species that had the most consistent and significant changes in expression following influenza A virus infection (Table 2 ) [20] .
TextSentencer_T61 9213-9466 Sentence denotes We then examined the list of targets predicted by TargetScan computer software (http://www.targetscan. org/) for the miRNA species that had the most consistent and significant changes in expression following influenza A virus infection (Table 2 ) [20] .
TextSentencer_T62 9467-9594 Sentence denotes The TargetScan results showed that many of the target genes were involved in the inflammatory response and cell death pathways.
TextSentencer_T62 9467-9594 Sentence denotes The TargetScan results showed that many of the target genes were involved in the inflammatory response and cell death pathways.
TextSentencer_T63 9595-9735 Sentence denotes Interestingly, one of the target prediction results showed that there was a 3′ untranslated region (UTR) binding site on TGF-β2 for miR-141.
TextSentencer_T63 9595-9735 Sentence denotes Interestingly, one of the target prediction results showed that there was a 3′ untranslated region (UTR) binding site on TGF-β2 for miR-141.
TextSentencer_T64 9736-9760 Sentence denotes The miR-141 sequence is:
TextSentencer_T64 9736-9760 Sentence denotes The miR-141 sequence is:
TextSentencer_T65 9761-9822 Sentence denotes 3′-GGUAGAAAUGGUCUGUCACAAU -5′, while that of TGF-β2 3′UTR is:
TextSentencer_T65 9761-9822 Sentence denotes 3′-GGUAGAAAUGGUCUGUCACAAU -5′, while that of TGF-β2 3′UTR is:
TextSentencer_T66 9823-9854 Sentence denotes 5′-AGAGCCUUGGUUC AUCAGUGUUA-3′.
TextSentencer_T66 9823-9854 Sentence denotes 5′-AGAGCCUUGGUUC AUCAGUGUUA-3′.
TextSentencer_T67 9855-10203 Sentence denotes We had previously reported that TGF-β2 was an important cytokine involved in the inflammatory response of avian influenza A virus infection [21] and, together with the results showing that the expression of miR-141 was altered during the time course of influenza A virus infection, we selected miR-141 for further functional analysis in this study.
TextSentencer_T67 9855-10203 Sentence denotes We had previously reported that TGF-β2 was an important cytokine involved in the inflammatory response of avian influenza A virus infection [21] and, together with the results showing that the expression of miR-141 was altered during the time course of influenza A virus infection, we selected miR-141 for further functional analysis in this study.
TextSentencer_T68 10204-10461 Sentence denotes In addition to the miRNA target prediction results, by using ecoptic expression of miR-141, the level of TGF-β2 mRNA was found to be significantly decreased in miR-141 transfected cells but not in negative-control miRNA mimic transfected cells ( Figure 2 ).
TextSentencer_T68 10204-10461 Sentence denotes In addition to the miRNA target prediction results, by using ecoptic expression of miR-141, the level of TGF-β2 mRNA was found to be significantly decreased in miR-141 transfected cells but not in negative-control miRNA mimic transfected cells ( Figure 2 ).
TextSentencer_T69 10462-10695 Sentence denotes In this overexpression system we could determine that the 3′UTR was the miR-141 target and the decreased TGF-β2 mRNA level might be due to the binding of miR-141 to the 3′UTR of TGF-β2 mRNA which reduced the halflives of TGF-β2 mRNA.
TextSentencer_T69 10462-10695 Sentence denotes In this overexpression system we could determine that the 3′UTR was the miR-141 target and the decreased TGF-β2 mRNA level might be due to the binding of miR-141 to the 3′UTR of TGF-β2 mRNA which reduced the halflives of TGF-β2 mRNA.
TextSentencer_T70 10696-10825 Sentence denotes The functional relevance of changes in miR-141 expression during influenza A virus infection was assessed using miRNA inhibitors.
TextSentencer_T70 10696-10825 Sentence denotes The functional relevance of changes in miR-141 expression during influenza A virus infection was assessed using miRNA inhibitors.
TextSentencer_T71 10826-10971 Sentence denotes Chemically modified, single stranded nucleic acids anti-miR miR-141 inhibitor and negative control were transfected into H292 cells for 24 hours.
TextSentencer_T71 10826-10971 Sentence denotes Chemically modified, single stranded nucleic acids anti-miR miR-141 inhibitor and negative control were transfected into H292 cells for 24 hours.
TextSentencer_T72 10972-11131 Sentence denotes We had previously shown that this was sufficient time to obtain oligonucleotide delivery in H292 cells when examining the inhibition of TGF-β2 mRNA expression.
TextSentencer_T72 10972-11131 Sentence denotes We had previously shown that this was sufficient time to obtain oligonucleotide delivery in H292 cells when examining the inhibition of TGF-β2 mRNA expression.
TextSentencer_T73 11132-11257 Sentence denotes After the cells were pre-treated with anti-miR miR-141 for 24 hours, they were then infected with H1N1 or H5N1, respectively.
TextSentencer_T73 11132-11257 Sentence denotes After the cells were pre-treated with anti-miR miR-141 for 24 hours, they were then infected with H1N1 or H5N1, respectively.
TextSentencer_T74 11258-11393 Sentence denotes After the infection processes, anti-miR miR-141 was transfected again into the virus infected cells and incubated for another 24 hours.
TextSentencer_T74 11258-11393 Sentence denotes After the infection processes, anti-miR miR-141 was transfected again into the virus infected cells and incubated for another 24 hours.
TextSentencer_T75 11394-11668 Sentence denotes The results of this experiment showed that the anti-miR miR-141 inhibitor could cause an increase in TGF-β2 protein expression in H1N1 or H5N1 infected cells, as compared to cells only infected with H1N1 or H5N1 but without anti-miR miR-141 inhibitor treatment ( Figure 3 ).
TextSentencer_T75 11394-11668 Sentence denotes The results of this experiment showed that the anti-miR miR-141 inhibitor could cause an increase in TGF-β2 protein expression in H1N1 or H5N1 infected cells, as compared to cells only infected with H1N1 or H5N1 but without anti-miR miR-141 inhibitor treatment ( Figure 3 ).
TextSentencer_T76 11669-11740 Sentence denotes The effect was also more prominent in H5N1 infection than that of H1N1.
TextSentencer_T76 11669-11740 Sentence denotes The effect was also more prominent in H5N1 infection than that of H1N1.
TextSentencer_T77 11741-11791 Sentence denotes Figure 2 The TGF-β2 3′UTR is regulated by miR-141.
TextSentencer_T77 11741-11791 Sentence denotes Figure 2 The TGF-β2 3′UTR is regulated by miR-141.
TextSentencer_T78 11792-11876 Sentence denotes NCI-H292 cells were transfected with pre-miR-141 and negative control, respectively.
TextSentencer_T78 11792-11876 Sentence denotes NCI-H292 cells were transfected with pre-miR-141 and negative control, respectively.
TextSentencer_T79 11877-11972 Sentence denotes The fold-changes of mRNA level of TGF-β2 as measured by qRT-PCR at 24 hours after transfection.
TextSentencer_T79 11877-11972 Sentence denotes The fold-changes of mRNA level of TGF-β2 as measured by qRT-PCR at 24 hours after transfection.
TextSentencer_T80 11973-12112 Sentence denotes Fold-changes were calculated by ΔΔCT method as compared with negatively transfected cell control and using β-actin level for normalization.
TextSentencer_T80 11973-12112 Sentence denotes Fold-changes were calculated by ΔΔCT method as compared with negatively transfected cell control and using β-actin level for normalization.
TextSentencer_T81 12113-12170 Sentence denotes Each point on the graph represents the mean fold-changes.
TextSentencer_T81 12113-12170 Sentence denotes Each point on the graph represents the mean fold-changes.
TextSentencer_T82 12171-12272 Sentence denotes The mean fold-changes of TGF-β2 mRNA level was compared to that of negative control ± SD (p* < 0.05).
TextSentencer_T82 12171-12272 Sentence denotes The mean fold-changes of TGF-β2 mRNA level was compared to that of negative control ± SD (p* < 0.05).
TextSentencer_T83 12273-12403 Sentence denotes In this study we examined the connection between influenza A virus infection and the global patterns of cellular miRNA expression.
TextSentencer_T83 12273-12403 Sentence denotes In this study we examined the connection between influenza A virus infection and the global patterns of cellular miRNA expression.
TextSentencer_T84 12404-12534 Sentence denotes The major observations from this work were that influenza A virus infection resulted in the altered regulation of cellular miRNAs.
TextSentencer_T84 12404-12534 Sentence denotes The major observations from this work were that influenza A virus infection resulted in the altered regulation of cellular miRNAs.
TextSentencer_T85 12535-12651 Sentence denotes Avian influenza A virus can alter cellular miRNAs to a greater extent than that of seasonal human influenza A virus.
TextSentencer_T85 12535-12651 Sentence denotes Avian influenza A virus can alter cellular miRNAs to a greater extent than that of seasonal human influenza A virus.
TextSentencer_T86 12652-12720 Sentence denotes Influenza A virus affects the regulation of many cellular processes.
TextSentencer_T86 12652-12720 Sentence denotes Influenza A virus affects the regulation of many cellular processes.
TextSentencer_T87 12721-12849 Sentence denotes In some cases, these changes are directed by the virus for its advantage and others are cellular defense responses to infection.
TextSentencer_T87 12721-12849 Sentence denotes In some cases, these changes are directed by the virus for its advantage and others are cellular defense responses to infection.
TextSentencer_T88 12850-12943 Sentence denotes Here, we found that influenza A virus infection led to altered regulation of cellular miRNAs.
TextSentencer_T88 12850-12943 Sentence denotes Here, we found that influenza A virus infection led to altered regulation of cellular miRNAs.
TextSentencer_T89 12944-13136 Sentence denotes Given the number of genes that can be regulated by individual miRNAs and the number of miRNAs expressed in cells, this greatly expands the range of possible virus-host regulatory interactions.
TextSentencer_T89 12944-13136 Sentence denotes Given the number of genes that can be regulated by individual miRNAs and the number of miRNAs expressed in cells, this greatly expands the range of possible virus-host regulatory interactions.
TextSentencer_T90 13137-13345 Sentence denotes The complexity is underscored by there being no uniform global pattern of regulation; rather, it appears that individual (or groups of ) miRNA are independently regulated, some positively and some negatively.
TextSentencer_T90 13137-13345 Sentence denotes The complexity is underscored by there being no uniform global pattern of regulation; rather, it appears that individual (or groups of ) miRNA are independently regulated, some positively and some negatively.
TextSentencer_T91 13346-13475 Sentence denotes Persistent and transient effects were seen, and changes in miRNA expression profiles were linked to the time course of infection.
TextSentencer_T91 13346-13475 Sentence denotes Persistent and transient effects were seen, and changes in miRNA expression profiles were linked to the time course of infection.
TextSentencer_T92 13476-13644 Sentence denotes As a summary, miR-1246, miR-663 and miR-574-3p were up-regulated (>3-fold, p<0.05) at 24-hour postinfection with subtype H5 as compared with non-infected control cells.
TextSentencer_T92 13476-13644 Sentence denotes As a summary, miR-1246, miR-663 and miR-574-3p were up-regulated (>3-fold, p<0.05) at 24-hour postinfection with subtype H5 as compared with non-infected control cells.
TextSentencer_T93 13645-13875 Sentence denotes Moreover, miR-100*, miR-21*, miR-141, miR-1274a and miR1274b were found to be downregulated (>3-fold, p<0.05) in infection with subtype H5, particularly at 18 or 24 hours post-infection as compared with non-infected control cells.
TextSentencer_T93 13645-13875 Sentence denotes Moreover, miR-100*, miR-21*, miR-141, miR-1274a and miR1274b were found to be downregulated (>3-fold, p<0.05) in infection with subtype H5, particularly at 18 or 24 hours post-infection as compared with non-infected control cells.
TextSentencer_T94 13876-14132 Sentence denotes Interestingly, many of the virally regulated miRNAs were predicted by TargetScan to target important biological pathways, immune-related signal pathways and have altered regulation in some cellular defense and some states of cellular differentiation [22] .
TextSentencer_T94 13876-14132 Sentence denotes Interestingly, many of the virally regulated miRNAs were predicted by TargetScan to target important biological pathways, immune-related signal pathways and have altered regulation in some cellular defense and some states of cellular differentiation [22] .
TextSentencer_T95 14133-14231 Sentence denotes In our study, we found that the expression of miR-141 was affected by influenza A virus infection.
TextSentencer_T95 14133-14231 Sentence denotes In our study, we found that the expression of miR-141 was affected by influenza A virus infection.
TextSentencer_T96 14232-14420 Sentence denotes To validate the in silico findings empirically on the target of miR-141, we checked whether transient-transfection of anti-and premir-141 into NCI-H292 cells resulted in TGF-β2 regulation.
TextSentencer_T96 14232-14420 Sentence denotes To validate the in silico findings empirically on the target of miR-141, we checked whether transient-transfection of anti-and premir-141 into NCI-H292 cells resulted in TGF-β2 regulation.
TextSentencer_T97 14421-14549 Sentence denotes In our experiment, the transfection efficiency was an important factor affecting the degree of regulation on the target gene(s).
TextSentencer_T97 14421-14549 Sentence denotes In our experiment, the transfection efficiency was an important factor affecting the degree of regulation on the target gene(s).
TextSentencer_T98 14550-14719 Sentence denotes In the case of higher transfection efficiency, as more miRNA would be transfected into the cells, the effect of gene(s) regulation by miRNA transfected would be greater.
TextSentencer_T98 14550-14719 Sentence denotes In the case of higher transfection efficiency, as more miRNA would be transfected into the cells, the effect of gene(s) regulation by miRNA transfected would be greater.
TextSentencer_T99 14720-14865 Sentence denotes In our study, the transfection efficiency was about 78.2 ± 6.3% (mean ± SD), which was considered to be adequate for further functional analyses.
TextSentencer_T99 14720-14865 Sentence denotes In our study, the transfection efficiency was about 78.2 ± 6.3% (mean ± SD), which was considered to be adequate for further functional analyses.
TextSentencer_T100 14866-15054 Sentence denotes During transfection, some oligonucleotide molecules were sequestered in internal vesicles and physically separated from their targets in the cytoplasm; and then released during cell lysis.
TextSentencer_T100 14866-15054 Sentence denotes During transfection, some oligonucleotide molecules were sequestered in internal vesicles and physically separated from their targets in the cytoplasm; and then released during cell lysis.
TextSentencer_T101 15055-15321 Sentence denotes Therefore monitoring miRNAs by qPCR after transfection qRT-PCR were used to quantitify the TGF-β2 mRNA levels and fold-changes were calculated by ΔΔCT method as compared with non-infection cell control (mock) and using endogeneous actin mRNA level for normalization.
TextSentencer_T101 15055-15321 Sentence denotes Therefore monitoring miRNAs by qPCR after transfection qRT-PCR were used to quantitify the TGF-β2 mRNA levels and fold-changes were calculated by ΔΔCT method as compared with non-infection cell control (mock) and using endogeneous actin mRNA level for normalization.
TextSentencer_T102 15322-15415 Sentence denotes TGF-β2 protein level was measured by enzyme-linked immunosorbent assay as compared with mock.
TextSentencer_T102 15322-15415 Sentence denotes TGF-β2 protein level was measured by enzyme-linked immunosorbent assay as compared with mock.
TextSentencer_T103 15416-15474 Sentence denotes Each point on the graph respresents the mean fold-changes.
TextSentencer_T103 15416-15474 Sentence denotes Each point on the graph respresents the mean fold-changes.
TextSentencer_T104 15475-15621 Sentence denotes The experimental mean fold-changes of mRNA and protein levels were compared to that of mock controls ± SD (p* < 0.05), (p # < 0.05), respectively.
TextSentencer_T104 15475-15621 Sentence denotes The experimental mean fold-changes of mRNA and protein levels were compared to that of mock controls ± SD (p* < 0.05), (p # < 0.05), respectively.
TextSentencer_T105 15622-15644 Sentence denotes would not be valuable.
TextSentencer_T105 15622-15644 Sentence denotes would not be valuable.
TextSentencer_T106 15645-15783 Sentence denotes Previous researchers of this procedure had highly recommended investigating the target mRNAs and proteins instead of miRNA quantification.
TextSentencer_T106 15645-15783 Sentence denotes Previous researchers of this procedure had highly recommended investigating the target mRNAs and proteins instead of miRNA quantification.
TextSentencer_T107 15784-16140 Sentence denotes The time point of 24-hour post-transfection or postinfection was chosen for evaluation because miR-141 induction was observed at the early stage of virus infection, and sufficient time might be required for the miR-141 to have effect on its target(s), so we had chosen 24-hour post-transfection or post-infection for evaluation of the effect of this miRNA.
TextSentencer_T107 15784-16140 Sentence denotes The time point of 24-hour post-transfection or postinfection was chosen for evaluation because miR-141 induction was observed at the early stage of virus infection, and sufficient time might be required for the miR-141 to have effect on its target(s), so we had chosen 24-hour post-transfection or post-infection for evaluation of the effect of this miRNA.
TextSentencer_T108 16141-16310 Sentence denotes Indeed, upon detecting the TGF-β2 expression at mRNA and protein levels, we found that the altered miR-141 expression would affect the expression of the cytokine-TGF-β2.
TextSentencer_T108 16141-16310 Sentence denotes Indeed, upon detecting the TGF-β2 expression at mRNA and protein levels, we found that the altered miR-141 expression would affect the expression of the cytokine-TGF-β2.
TextSentencer_T109 16311-16467 Sentence denotes Literature search on the background of miR-141 confirmed that miR-141 is a member of the miR-200 family (miR-200a, miR-200b, miR-200c, miR-141 and miR-429).
TextSentencer_T109 16311-16467 Sentence denotes Literature search on the background of miR-141 confirmed that miR-141 is a member of the miR-200 family (miR-200a, miR-200b, miR-200c, miR-141 and miR-429).
TextSentencer_T110 16468-16530 Sentence denotes Previous studies of miR-141 were mainly on its role in cancer.
TextSentencer_T110 16468-16530 Sentence denotes Previous studies of miR-141 were mainly on its role in cancer.
TextSentencer_T111 16531-16668 Sentence denotes It has been reported that miR-141 were markedly downregulated in cells that had undergone epithelial to mesenchymal in response to TGF-β.
TextSentencer_T111 16531-16668 Sentence denotes It has been reported that miR-141 were markedly downregulated in cells that had undergone epithelial to mesenchymal in response to TGF-β.
TextSentencer_T112 16669-16951 Sentence denotes MiR-141 was also found to be overexpressed in ovarian and colorectal cancers [23, 24] and downregulated in prostate, hepatocellular, renal cell carcinoma and in gastric cancer tissues [25] [26] [27] [28] raising a controversial issue about the role of miR-141 in cancer progression.
TextSentencer_T112 16669-16951 Sentence denotes MiR-141 was also found to be overexpressed in ovarian and colorectal cancers [23, 24] and downregulated in prostate, hepatocellular, renal cell carcinoma and in gastric cancer tissues [25] [26] [27] [28] raising a controversial issue about the role of miR-141 in cancer progression.
TextSentencer_T113 16952-17065 Sentence denotes Furthermore, the miR-200 family members play roles in maintaining the epithelial phenotype of cancer cells [29] .
TextSentencer_T113 16952-17065 Sentence denotes Furthermore, the miR-200 family members play roles in maintaining the epithelial phenotype of cancer cells [29] .
TextSentencer_T114 17066-17210 Sentence denotes A member of this family -miR-200a was also found to be differentially expressed in response to influenza virus infection in another study [17] .
TextSentencer_T114 17066-17210 Sentence denotes A member of this family -miR-200a was also found to be differentially expressed in response to influenza virus infection in another study [17] .
TextSentencer_T115 17211-17398 Sentence denotes The targets of miR-200a are associated with viral gene replication and the JAK-STAT signaling pathway, which is closely related to type I interferon-mediated innate immune response [17] .
TextSentencer_T115 17211-17398 Sentence denotes The targets of miR-200a are associated with viral gene replication and the JAK-STAT signaling pathway, which is closely related to type I interferon-mediated innate immune response [17] .
TextSentencer_T116 17399-17650 Sentence denotes However, the effect of miR-141 on virus infection was not known, except one recent report showing that enterovirus can induce miR-141 and contribute to the shutoff of host protein translation by targeting the translation initiation factor eIF4E [30] .
TextSentencer_T116 17399-17650 Sentence denotes However, the effect of miR-141 on virus infection was not known, except one recent report showing that enterovirus can induce miR-141 and contribute to the shutoff of host protein translation by targeting the translation initiation factor eIF4E [30] .
TextSentencer_T117 17651-17797 Sentence denotes In addition, evidence suggests that influenza A virus infection reduces or promotes the expression of the host miR-141 in a time dependent manner.
TextSentencer_T117 17651-17797 Sentence denotes In addition, evidence suggests that influenza A virus infection reduces or promotes the expression of the host miR-141 in a time dependent manner.
TextSentencer_T118 17798-17870 Sentence denotes We found that TGF-β2 mRNA was suppressed in miR-141 overexpressed cells.
TextSentencer_T118 17798-17870 Sentence denotes We found that TGF-β2 mRNA was suppressed in miR-141 overexpressed cells.
TextSentencer_T119 17871-18093 Sentence denotes Our observation is in line with another study showing that the 3′UTR of TGF-β2 mRNA contained a target site for miR-141/200a and the expression of TGF-β2 was significantly decreased in miR-141/200a transfected cells [22] .
TextSentencer_T119 17871-18093 Sentence denotes Our observation is in line with another study showing that the 3′UTR of TGF-β2 mRNA contained a target site for miR-141/200a and the expression of TGF-β2 was significantly decreased in miR-141/200a transfected cells [22] .
TextSentencer_T120 18094-18257 Sentence denotes Furthermore, miR-141 may not only work as translational repressors of target mRNAs, because it was observed that they also caused a decrease in TGF-β2 mRNA levels.
TextSentencer_T120 18094-18257 Sentence denotes Furthermore, miR-141 may not only work as translational repressors of target mRNAs, because it was observed that they also caused a decrease in TGF-β2 mRNA levels.
TextSentencer_T121 18258-18379 Sentence denotes These findings are similar to recent data demonstrating that some miRNAs can alter the mRNA levels of target genes [31] .
TextSentencer_T121 18258-18379 Sentence denotes These findings are similar to recent data demonstrating that some miRNAs can alter the mRNA levels of target genes [31] .
TextSentencer_T122 18380-18498 Sentence denotes This ability is probably independent of the ability of these miRNAs to regulate the translation of target mRNAs [14] .
TextSentencer_T122 18380-18498 Sentence denotes This ability is probably independent of the ability of these miRNAs to regulate the translation of target mRNAs [14] .
TextSentencer_T123 18499-18621 Sentence denotes We also noted that antagomiR-141 moderately increased the accumulation of TGF-β2 protein during influenza virus infection.
TextSentencer_T123 18499-18621 Sentence denotes We also noted that antagomiR-141 moderately increased the accumulation of TGF-β2 protein during influenza virus infection.
TextSentencer_T124 18622-18881 Sentence denotes This might be because, by the use of anti-miR miR-141 inhibitor, which decreases the cellular pool of miR-141, the translation control of the TGF-β2 mRNA was subsequently released and caused the TGF-β2 protein to express and accumulate during virus infection.
TextSentencer_T124 18622-18881 Sentence denotes This might be because, by the use of anti-miR miR-141 inhibitor, which decreases the cellular pool of miR-141, the translation control of the TGF-β2 mRNA was subsequently released and caused the TGF-β2 protein to express and accumulate during virus infection.
TextSentencer_T125 18882-19108 Sentence denotes However, it was also observed that when there was an increase in TGF-β2 mRNA level, the corresponding TGF-β2 protein expression level would be increased, except in the case of non-miR-141-inhibitor treated H5N1 infected cells.
TextSentencer_T125 18882-19108 Sentence denotes However, it was also observed that when there was an increase in TGF-β2 mRNA level, the corresponding TGF-β2 protein expression level would be increased, except in the case of non-miR-141-inhibitor treated H5N1 infected cells.
TextSentencer_T126 19109-19205 Sentence denotes In this case, there was a decrease in TGF-β2 mRNA level, while the TGF-β2 protein was increased.
TextSentencer_T126 19109-19205 Sentence denotes In this case, there was a decrease in TGF-β2 mRNA level, while the TGF-β2 protein was increased.
TextSentencer_T127 19206-19362 Sentence denotes This might be explained by the fact that TGF-β2 mRNA degradation induced by miR-141 might be much faster than that of the corresponding protein degradation.
TextSentencer_T127 19206-19362 Sentence denotes This might be explained by the fact that TGF-β2 mRNA degradation induced by miR-141 might be much faster than that of the corresponding protein degradation.
TextSentencer_T128 19363-19497 Sentence denotes Recently, we had also reported that H1N1 was the only subtype that could induce a sustained increase in TGF-β2 at protein level [21] .
TextSentencer_T128 19363-19497 Sentence denotes Recently, we had also reported that H1N1 was the only subtype that could induce a sustained increase in TGF-β2 at protein level [21] .
TextSentencer_T129 19498-19731 Sentence denotes That observation coincides with our results in this study, showing that H1N1 infection induced a little amount of miR-141 expression, while H5N1 infection induced a higher amount of miR-141 expression at the early phase of infection.
TextSentencer_T129 19498-19731 Sentence denotes That observation coincides with our results in this study, showing that H1N1 infection induced a little amount of miR-141 expression, while H5N1 infection induced a higher amount of miR-141 expression at the early phase of infection.
TextSentencer_T130 19732-19876 Sentence denotes As a consequence of the higher amount of miR-141 in H5N1 infection, TGF-β2 expression might be more greatly reduced than that in H1N1 infection.
TextSentencer_T130 19732-19876 Sentence denotes As a consequence of the higher amount of miR-141 in H5N1 infection, TGF-β2 expression might be more greatly reduced than that in H1N1 infection.
TextSentencer_T131 19877-20082 Sentence denotes Since TGF-β2 can act as both an immunosuppressive agent and a potent proinflammatory molecule through its ability to attract and regulate inflammatory molecules, it plays a vital role in T-cell inhibition.
TextSentencer_T131 19877-20082 Sentence denotes Since TGF-β2 can act as both an immunosuppressive agent and a potent proinflammatory molecule through its ability to attract and regulate inflammatory molecules, it plays a vital role in T-cell inhibition.
TextSentencer_T132 20083-20281 Sentence denotes Furthermore, it has been reported that TGF-β2 inhibits Th1 cytokine-mediated induction of CCL-2/MCP-1, CCL-3/MIP-1α, CCL-4/MIP-1β, CCL-5/RANTES, CCL-9 /MIP-1γ, CXCL-2/MIP-2, and CXCL-10/IP-10 [32] .
TextSentencer_T132 20083-20281 Sentence denotes Furthermore, it has been reported that TGF-β2 inhibits Th1 cytokine-mediated induction of CCL-2/MCP-1, CCL-3/MIP-1α, CCL-4/MIP-1β, CCL-5/RANTES, CCL-9 /MIP-1γ, CXCL-2/MIP-2, and CXCL-10/IP-10 [32] .
TextSentencer_T133 20282-20547 Sentence denotes Moreover, the pro-inflammatory responses during influenza A virus infection are tightly controlled by anti-inflammatory mediators, such as TGF-β2, to protect the easily damageable lung tissue from destructive side effects associated with virus induced inflammation.
TextSentencer_T133 20282-20547 Sentence denotes Moreover, the pro-inflammatory responses during influenza A virus infection are tightly controlled by anti-inflammatory mediators, such as TGF-β2, to protect the easily damageable lung tissue from destructive side effects associated with virus induced inflammation.
TextSentencer_T134 20548-20741 Sentence denotes Therefore, the downregulation of TGF-β2 protein by miR-141 may be an important step in the excessive inflammation progression during influenza A virus infection, particularly in H5N1 infection.
TextSentencer_T134 20548-20741 Sentence denotes Therefore, the downregulation of TGF-β2 protein by miR-141 may be an important step in the excessive inflammation progression during influenza A virus infection, particularly in H5N1 infection.
TextSentencer_T135 20742-20910 Sentence denotes However, whether the recovery of TGF-β2 expression by anti-miR miR-141 inhibitor could resolve the hypercytokinemia stage of H5N1 infection needs to be further studied.
TextSentencer_T135 20742-20910 Sentence denotes However, whether the recovery of TGF-β2 expression by anti-miR miR-141 inhibitor could resolve the hypercytokinemia stage of H5N1 infection needs to be further studied.
TextSentencer_T136 20911-21078 Sentence denotes Although our findings were obtained from an in vitro model, we could apply these to the real situation of an in vivo model or tissue comprised of different cell types.
TextSentencer_T136 20911-21078 Sentence denotes Although our findings were obtained from an in vitro model, we could apply these to the real situation of an in vivo model or tissue comprised of different cell types.
TextSentencer_T137 21079-21183 Sentence denotes In real bronchial environments, lung epithelial cells are the key target of influenza viruses [33, 34] .
TextSentencer_T137 21079-21183 Sentence denotes In real bronchial environments, lung epithelial cells are the key target of influenza viruses [33, 34] .
TextSentencer_T138 21184-21366 Sentence denotes After these cells are infected, they will activate an inflammatory cascade which launches a quick antimicrobial reaction and directs adaptive immunity to mount a protective response.
TextSentencer_T138 21184-21366 Sentence denotes After these cells are infected, they will activate an inflammatory cascade which launches a quick antimicrobial reaction and directs adaptive immunity to mount a protective response.
TextSentencer_T139 21367-21528 Sentence denotes Bronchial epithelial cells therefore modulate the activation of monocytes, macrophages, dendritic cells (DC), and T lymphocytes through cytokines and chemokines.
TextSentencer_T139 21367-21528 Sentence denotes Bronchial epithelial cells therefore modulate the activation of monocytes, macrophages, dendritic cells (DC), and T lymphocytes through cytokines and chemokines.
TextSentencer_T140 21529-21658 Sentence denotes Cytokines and chemokines generally function in an autocrine (on the producing cell itself) or paracrine (on nearby cells) manner.
TextSentencer_T140 21529-21658 Sentence denotes Cytokines and chemokines generally function in an autocrine (on the producing cell itself) or paracrine (on nearby cells) manner.
TextSentencer_T141 21659-21824 Sentence denotes These mediators will contribute to the generation of a specific bronchial homeostatic microenvironment that affects the way in which the body copes with the viruses.
TextSentencer_T141 21659-21824 Sentence denotes These mediators will contribute to the generation of a specific bronchial homeostatic microenvironment that affects the way in which the body copes with the viruses.
TextSentencer_T142 21825-21918 Sentence denotes This homeostatic "circuit" can inhibit excessive inflammatory response in lung tissues [35] .
TextSentencer_T142 21825-21918 Sentence denotes This homeostatic "circuit" can inhibit excessive inflammatory response in lung tissues [35] .
TextSentencer_T143 21919-22036 Sentence denotes For example, TGF-β had been reported to mediate a cross-talk between alveolar macrophages and epithelial cells [36] .
TextSentencer_T143 21919-22036 Sentence denotes For example, TGF-β had been reported to mediate a cross-talk between alveolar macrophages and epithelial cells [36] .
TextSentencer_T144 22037-22172 Sentence denotes However, our findings show that, during highly pathogenic H5N1 avian virus infection, miR-141 would be induced shortly after infection.
TextSentencer_T144 22037-22172 Sentence denotes However, our findings show that, during highly pathogenic H5N1 avian virus infection, miR-141 would be induced shortly after infection.
TextSentencer_T145 22173-22276 Sentence denotes With high level of miR-141, the expression of TGF-β would be suppressed from the lung epithelial cells.
TextSentencer_T145 22173-22276 Sentence denotes With high level of miR-141, the expression of TGF-β would be suppressed from the lung epithelial cells.
TextSentencer_T146 22277-22422 Sentence denotes Without sufficient TGF-β, the pro-inflammatory response might not be tightly controlled in cases of highly pathogenic H5N1 avian virus infection.
TextSentencer_T146 22277-22422 Sentence denotes Without sufficient TGF-β, the pro-inflammatory response might not be tightly controlled in cases of highly pathogenic H5N1 avian virus infection.
TextSentencer_T147 22423-22643 Sentence denotes This might explain the mechanism concerning bronchial infiltration of inflammatory cells, particularly lymphocytes and eosinophils, and the subsequent hyperresponsiveness of the bronchial wall induced by viral infection.
TextSentencer_T147 22423-22643 Sentence denotes This might explain the mechanism concerning bronchial infiltration of inflammatory cells, particularly lymphocytes and eosinophils, and the subsequent hyperresponsiveness of the bronchial wall induced by viral infection.
TextSentencer_T148 22644-22724 Sentence denotes Our study has some limitations that will need to be addressed in future studies.
TextSentencer_T148 22644-22724 Sentence denotes Our study has some limitations that will need to be addressed in future studies.
TextSentencer_T149 22725-22829 Sentence denotes Firstly, we did not assess the roles of other miRNAs whose expression were also altered after infection.
TextSentencer_T149 22725-22829 Sentence denotes Firstly, we did not assess the roles of other miRNAs whose expression were also altered after infection.
TextSentencer_T150 22830-23061 Sentence denotes The miRNA microarrays that we used did not contain probes for every known miRNA; thus it is possible that influenza A virus infection affects the expression of some other miRNAs not yet covered by the kit used in the current study.
TextSentencer_T150 22830-23061 Sentence denotes The miRNA microarrays that we used did not contain probes for every known miRNA; thus it is possible that influenza A virus infection affects the expression of some other miRNAs not yet covered by the kit used in the current study.
TextSentencer_T151 23062-23202 Sentence denotes Secondly, the virus may interact with miRNA regulatory pathways differently in other cell or tissue types, or in other physiological status.
TextSentencer_T151 23062-23202 Sentence denotes Secondly, the virus may interact with miRNA regulatory pathways differently in other cell or tissue types, or in other physiological status.
TextSentencer_T152 23203-23387 Sentence denotes In conclusion, based on the broad-catching miRNA microarray approach, we found that dysregulation of miRNA expression is mainly observed in highly pathogenic avian influenza infection.
TextSentencer_T152 23203-23387 Sentence denotes In conclusion, based on the broad-catching miRNA microarray approach, we found that dysregulation of miRNA expression is mainly observed in highly pathogenic avian influenza infection.
TextSentencer_T153 23388-23481 Sentence denotes We identified that miR-141 was induced at early time points upon influenza A virus infection.
TextSentencer_T153 23388-23481 Sentence denotes We identified that miR-141 was induced at early time points upon influenza A virus infection.
TextSentencer_T154 23482-23562 Sentence denotes The induction was higher in H5N1 infection than that of seasonal H1N1 infection.
TextSentencer_T154 23482-23562 Sentence denotes The induction was higher in H5N1 infection than that of seasonal H1N1 infection.
TextSentencer_T155 23563-23695 Sentence denotes Moreover, TGF-β2, which plays an important role in regulating inflammatory processes, was identified as a target of miR-141 binding.
TextSentencer_T155 23563-23695 Sentence denotes Moreover, TGF-β2, which plays an important role in regulating inflammatory processes, was identified as a target of miR-141 binding.
TextSentencer_T156 23696-23838 Sentence denotes As a result, influenza A virus infection, in particular highly pathogenic H5N1, could affect the inflammatory processes via miR-141 induction.
TextSentencer_T156 23696-23838 Sentence denotes As a result, influenza A virus infection, in particular highly pathogenic H5N1, could affect the inflammatory processes via miR-141 induction.
TextSentencer_T157 23839-23842 Sentence denotes The
TextSentencer_T157 23839-23842 Sentence denotes The
TextSentencer_T158 23844-24232 Sentence denotes The bronchial epithelial cells -NCI-H292, derived from human lung mucoepidermoid carcinoma cells (ATCC, CRL-1848, Rockville, MD, USA), were grown as monolayers in RPMI-1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/mL streptomycin (all from Gibco, Life Technology, Rockville, Md., USA) at 37°C in a 5% CO 2 incubator.
TextSentencer_T158 23844-24232 Sentence denotes The bronchial epithelial cells -NCI-H292, derived from human lung mucoepidermoid carcinoma cells (ATCC, CRL-1848, Rockville, MD, USA), were grown as monolayers in RPMI-1640 medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/mL streptomycin (all from Gibco, Life Technology, Rockville, Md., USA) at 37°C in a 5% CO 2 incubator.
TextSentencer_T159 24233-24330 Sentence denotes NCI-H292 cells were used as an invitro model to study host cellular responses to viral infection.
TextSentencer_T159 24233-24330 Sentence denotes NCI-H292 cells were used as an invitro model to study host cellular responses to viral infection.
TextSentencer_T160 24331-24428 Sentence denotes Mandin-Darby canine kidney (MDCK) cells were used for growing stocks of influenza virus isolates.
TextSentencer_T160 24331-24428 Sentence denotes Mandin-Darby canine kidney (MDCK) cells were used for growing stocks of influenza virus isolates.
TextSentencer_T161 24429-24606 Sentence denotes MDCK cells were grown and maintained in Eagles Minimal Essential Media (MEM) containing 2% FBS, 100 U/ml penicillin and 100 μg/mL streptomycin (all from Gibco, Life Technology).
TextSentencer_T161 24429-24606 Sentence denotes MDCK cells were grown and maintained in Eagles Minimal Essential Media (MEM) containing 2% FBS, 100 U/ml penicillin and 100 μg/mL streptomycin (all from Gibco, Life Technology).
TextSentencer_T162 24607-24768 Sentence denotes NCI-H292 cells were grown to confluence in sterile T75 tissue culture flasks for the inoculation of virus isolate at a multiplicity of infection (m.o.i.) of one.
TextSentencer_T162 24607-24768 Sentence denotes NCI-H292 cells were grown to confluence in sterile T75 tissue culture flasks for the inoculation of virus isolate at a multiplicity of infection (m.o.i.) of one.
TextSentencer_T163 24769-25086 Sentence denotes After 1 hour of absorption, the virus was removed and 2 ml of fresh RPMI-1640 media with 2% FBS, 100 U/ml penicillin, 100 μg/mL streptomycin and 1μg/ml L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (all from Gibco, Life Technology) was added, and incubated at 37°C in 5% CO 2 humidified air.
TextSentencer_T163 24769-25086 Sentence denotes After 1 hour of absorption, the virus was removed and 2 ml of fresh RPMI-1640 media with 2% FBS, 100 U/ml penicillin, 100 μg/mL streptomycin and 1μg/ml L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (all from Gibco, Life Technology) was added, and incubated at 37°C in 5% CO 2 humidified air.
TextSentencer_T164 25087-25223 Sentence denotes Total RNA was extracted from normal and infected NCI-H292 cells using Trizol reagent (Invitrogen) following the manufacturer's protocol.
TextSentencer_T164 25087-25223 Sentence denotes Total RNA was extracted from normal and infected NCI-H292 cells using Trizol reagent (Invitrogen) following the manufacturer's protocol.
TextSentencer_T165 25224-25273 Sentence denotes RNA pellets were resuspended in RNase-free water.
TextSentencer_T165 25224-25273 Sentence denotes RNA pellets were resuspended in RNase-free water.
TextSentencer_T166 25274-25379 Sentence denotes The RNA integrity was assessed using Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).
TextSentencer_T166 25274-25379 Sentence denotes The RNA integrity was assessed using Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).
TextSentencer_T167 25380-25527 Sentence denotes MiRNAs were labeled using the Agilent miRNA labeling reagent and hybridized to Agilent human miRNA arrays according to the manufacturer's protocol.
TextSentencer_T167 25380-25527 Sentence denotes MiRNAs were labeled using the Agilent miRNA labeling reagent and hybridized to Agilent human miRNA arrays according to the manufacturer's protocol.
TextSentencer_T168 25528-25633 Sentence denotes Briefly, total RNA (100 ng) was dephosphorylated and ligated with 3′, 5′-cytidine bisphosphate (pCp-Cy3).
TextSentencer_T168 25528-25633 Sentence denotes Briefly, total RNA (100 ng) was dephosphorylated and ligated with 3′, 5′-cytidine bisphosphate (pCp-Cy3).
TextSentencer_T169 25634-25802 Sentence denotes Labeled RNA was purified and hybridized to Agilent miRNA arrays with eight identical arrays per slide, with each array containing probes interrogating 866 human miRNAs.
TextSentencer_T169 25634-25802 Sentence denotes Labeled RNA was purified and hybridized to Agilent miRNA arrays with eight identical arrays per slide, with each array containing probes interrogating 866 human miRNAs.
TextSentencer_T170 25803-25970 Sentence denotes Images were scanned with the Agilent microarray scanner (Agilent Technologies), gridded, and analyzed using Agilent feature extraction software (Agilent Technologies).
TextSentencer_T170 25803-25970 Sentence denotes Images were scanned with the Agilent microarray scanner (Agilent Technologies), gridded, and analyzed using Agilent feature extraction software (Agilent Technologies).
TextSentencer_T171 25971-26113 Sentence denotes The cells were infected with either (A) the H1N1/2002 strain or (B) the H5N1/2004 strain, or (C) mockinfected with PBS (no infection control).
TextSentencer_T171 25971-26113 Sentence denotes The cells were infected with either (A) the H1N1/2002 strain or (B) the H5N1/2004 strain, or (C) mockinfected with PBS (no infection control).
TextSentencer_T172 26114-26182 Sentence denotes Cell samples were collected at 3, 6, 18 and 24 hours post-infection.
TextSentencer_T172 26114-26182 Sentence denotes Cell samples were collected at 3, 6, 18 and 24 hours post-infection.
TextSentencer_T173 26183-26243 Sentence denotes Each miRNA array allowed us to interrogate 866 human miRNAs.
TextSentencer_T173 26183-26243 Sentence denotes Each miRNA array allowed us to interrogate 866 human miRNAs.
TextSentencer_T174 26244-26329 Sentence denotes The results were analyzed using Genespring GX 10.0.2 software (Agilent Technologies).
TextSentencer_T174 26244-26329 Sentence denotes The results were analyzed using Genespring GX 10.0.2 software (Agilent Technologies).
TextSentencer_T175 26330-26387 Sentence denotes Firstly, the 16 arrays were quantile normalized together.
TextSentencer_T175 26330-26387 Sentence denotes Firstly, the 16 arrays were quantile normalized together.
TextSentencer_T176 26388-26665 Sentence denotes Then, student's paired t-test was applied to test if there was a significant difference between (A) the H1N1/2002-infected and (C) mock-infected, no infection control (matched for the time post-infection), (B) the H5N1/2004-infected and (C) mock-infected control, respectively.
TextSentencer_T176 26388-26665 Sentence denotes Then, student's paired t-test was applied to test if there was a significant difference between (A) the H1N1/2002-infected and (C) mock-infected, no infection control (matched for the time post-infection), (B) the H5N1/2004-infected and (C) mock-infected control, respectively.
TextSentencer_T177 26666-26801 Sentence denotes The resultant P-values were adjusted for multiple testing by using the Benjamini-Hochberg correction of the false-discovery rate [37] .
TextSentencer_T177 26666-26801 Sentence denotes The resultant P-values were adjusted for multiple testing by using the Benjamini-Hochberg correction of the false-discovery rate [37] .
TextSentencer_T178 26802-26888 Sentence denotes MiRNAs with this adjusted P-value <= 0.05 were considered as differentially expressed.
TextSentencer_T178 26802-26888 Sentence denotes MiRNAs with this adjusted P-value <= 0.05 were considered as differentially expressed.
TextSentencer_T179 26889-27023 Sentence denotes Those miRNAs, that are more than or equal to 3.5-fold up or down regulated were subjected to a second analysis using real-time RT-PCR.
TextSentencer_T179 26889-27023 Sentence denotes Those miRNAs, that are more than or equal to 3.5-fold up or down regulated were subjected to a second analysis using real-time RT-PCR.
TextSentencer_T180 27024-27186 Sentence denotes The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus [38] and are accessible through GEO Series accession number GSE44455.
TextSentencer_T180 27024-27186 Sentence denotes The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus [38] and are accessible through GEO Series accession number GSE44455.
TextSentencer_T181 27187-27337 Sentence denotes Total RNA was reverse transcribed with looped miRNAspecific RT primers contained in the TaqMan MicroRNA assays ((Applied Biosystems, Foster City, CA).
TextSentencer_T181 27187-27337 Sentence denotes Total RNA was reverse transcribed with looped miRNAspecific RT primers contained in the TaqMan MicroRNA assays ((Applied Biosystems, Foster City, CA).
TextSentencer_T182 27338-27540 Sentence denotes Briefly, single-stranded cDNA was synthesized from 10 ng total RNA in 15-μL reaction volume with TaqMan MicroRNA reverse transcription kit (Applied Biosystems), according to the manufacturer's protocol.
TextSentencer_T182 27338-27540 Sentence denotes Briefly, single-stranded cDNA was synthesized from 10 ng total RNA in 15-μL reaction volume with TaqMan MicroRNA reverse transcription kit (Applied Biosystems), according to the manufacturer's protocol.
TextSentencer_T183 27541-27649 Sentence denotes The reaction was incubated at 16°C for 30 min followed by 30 min at 42°C and inactivation at 85°C for 5 min.
TextSentencer_T183 27541-27649 Sentence denotes The reaction was incubated at 16°C for 30 min followed by 30 min at 42°C and inactivation at 85°C for 5 min.
TextSentencer_T184 27650-27741 Sentence denotes Each cDNA was amplified with sequence-specific TaqMan microRNA assays (Applied Biosystems).
TextSentencer_T184 27650-27741 Sentence denotes Each cDNA was amplified with sequence-specific TaqMan microRNA assays (Applied Biosystems).
TextSentencer_T185 27742-27928 Sentence denotes PCR reactions were performed on an Applied Biosystems Step One sequence detection system in 10 μl volumes at 95°C for 10 min, followed by 40 cycles of 95°C for 15 sec and 60°C for 1 min.
TextSentencer_T185 27742-27928 Sentence denotes PCR reactions were performed on an Applied Biosystems Step One sequence detection system in 10 μl volumes at 95°C for 10 min, followed by 40 cycles of 95°C for 15 sec and 60°C for 1 min.
TextSentencer_T186 27929-27967 Sentence denotes All samples were tested in triplicate.
TextSentencer_T186 27929-27967 Sentence denotes All samples were tested in triplicate.
TextSentencer_T187 27968-28169 Sentence denotes The threshold cycle (Ct) values obtained with the SDS software (Applied Biosystems) were compared with the Ct obtained from 18S rRNA assay (Applied Biosystems) for the normalization of total RNA input.
TextSentencer_T187 27968-28169 Sentence denotes The threshold cycle (Ct) values obtained with the SDS software (Applied Biosystems) were compared with the Ct obtained from 18S rRNA assay (Applied Biosystems) for the normalization of total RNA input.
TextSentencer_T188 28170-28270 Sentence denotes The fold-change was calculated based on Ct changes of mean medium Ct minus individual Ct of a miRNA.
TextSentencer_T188 28170-28270 Sentence denotes The fold-change was calculated based on Ct changes of mean medium Ct minus individual Ct of a miRNA.
TextSentencer_T189 28271-28315 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T189 28271-28315 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T190 28316-28505 Sentence denotes Total RNA extracted from cell cultures was reversely transcripted to cDNA using the poly(dT) primers and Superscript III reverse transcriptase (Invitrogen), and quantified by real-time PCR.
TextSentencer_T190 28316-28505 Sentence denotes Total RNA extracted from cell cultures was reversely transcripted to cDNA using the poly(dT) primers and Superscript III reverse transcriptase (Invitrogen), and quantified by real-time PCR.
TextSentencer_T191 28506-28596 Sentence denotes The sense and antisense primers used in real-time PCR for measuring TGF-β2 were: (Forward:
TextSentencer_T191 28506-28596 Sentence denotes The sense and antisense primers used in real-time PCR for measuring TGF-β2 were: (Forward:
TextSentencer_T192 28597-28633 Sentence denotes 5′-CCAAAGGGTACAATGCCAAC-3′; Reverse:
TextSentencer_T192 28597-28633 Sentence denotes 5′-CCAAAGGGTACAATGCCAAC-3′; Reverse:
TextSentencer_T193 28634-28662 Sentence denotes 5′-TAAGCTCAGGACCCTGCTGT-3′).
TextSentencer_T193 28634-28662 Sentence denotes 5′-TAAGCTCAGGACCCTGCTGT-3′).
TextSentencer_T194 28663-28778 Sentence denotes The realtime PCR reactions were performed in triplicates using the SYBER Green PCR Master Mix (Applied Biosystems).
TextSentencer_T194 28663-28778 Sentence denotes The realtime PCR reactions were performed in triplicates using the SYBER Green PCR Master Mix (Applied Biosystems).
TextSentencer_T195 28779-28898 Sentence denotes The PCR conditions were 95°C for 5 min, followed by 50 cycles of 95°C for 30 sec, 55°C for 30 sec, and 72°C for 30 sec.
TextSentencer_T195 28779-28898 Sentence denotes The PCR conditions were 95°C for 5 min, followed by 50 cycles of 95°C for 30 sec, 55°C for 30 sec, and 72°C for 30 sec.
TextSentencer_T196 28899-28985 Sentence denotes The expression of β-actin gene was also quantified in a similar way for normalization.
TextSentencer_T196 28899-28985 Sentence denotes The expression of β-actin gene was also quantified in a similar way for normalization.
TextSentencer_T197 28986-29185 Sentence denotes Comparative deltadelta C T method was used to analyze the results where expression level of the respective gene at the corresponding time point in non-transfected cells was regarded as one [39, 40] .
TextSentencer_T197 28986-29185 Sentence denotes Comparative deltadelta C T method was used to analyze the results where expression level of the respective gene at the corresponding time point in non-transfected cells was regarded as one [39, 40] .
TextSentencer_T198 29186-29230 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T198 29186-29230 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T199 29231-29334 Sentence denotes Cell culture supernatant was collected at 24 hours postinfection for the analysis of TGF-β2 expression.
TextSentencer_T199 29231-29334 Sentence denotes Cell culture supernatant was collected at 24 hours postinfection for the analysis of TGF-β2 expression.
TextSentencer_T200 29335-29513 Sentence denotes The total TGF-β2 protein level was measured by enzyme-linked immunosorbent assay (Emax® ImmunoAssay System, Promega, Madison, WI, USA) according to the manufacturer's procedures.
TextSentencer_T200 29335-29513 Sentence denotes The total TGF-β2 protein level was measured by enzyme-linked immunosorbent assay (Emax® ImmunoAssay System, Promega, Madison, WI, USA) according to the manufacturer's procedures.
TextSentencer_T201 29514-29558 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T201 29514-29558 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T202 29559-29691 Sentence denotes The cells were transfected in suspension after trypsinisation with 60 nM anti-miR, pre-miR or negative control (Applied Biosystems).
TextSentencer_T202 29559-29691 Sentence denotes The cells were transfected in suspension after trypsinisation with 60 nM anti-miR, pre-miR or negative control (Applied Biosystems).
TextSentencer_T203 29692-29772 Sentence denotes For the assay, 1x10 5 cells per mL were transfected per well of a 24-well plate.
TextSentencer_T203 29692-29772 Sentence denotes For the assay, 1x10 5 cells per mL were transfected per well of a 24-well plate.
TextSentencer_T204 29773-29915 Sentence denotes Transfection complexes were prepared in OptiMEM (Invitrogen) with 1.5 μL/24-well of siPORT NeoFx transfection agent (Ambion, Austin, TX, USA).
TextSentencer_T204 29773-29915 Sentence denotes Transfection complexes were prepared in OptiMEM (Invitrogen) with 1.5 μL/24-well of siPORT NeoFx transfection agent (Ambion, Austin, TX, USA).
TextSentencer_T205 29916-30033 Sentence denotes At 24 hours posttransfection, the cells were lysed for qRT-PCR analysis or subjected to H1N1 or H5N1 virus infection.
TextSentencer_T205 29916-30033 Sentence denotes At 24 hours posttransfection, the cells were lysed for qRT-PCR analysis or subjected to H1N1 or H5N1 virus infection.
TextSentencer_T206 30034-30280 Sentence denotes The transfection efficiency was calculated from the percentage of fluorescent cells that were observed using florescence microscopy after the transfection of fluorescein isothiocyanate (FITC)-labeled short nucleotide primers in separate controls.
TextSentencer_T206 30034-30280 Sentence denotes The transfection efficiency was calculated from the percentage of fluorescent cells that were observed using florescence microscopy after the transfection of fluorescein isothiocyanate (FITC)-labeled short nucleotide primers in separate controls.
TextSentencer_T207 30281-30408 Sentence denotes The transfection efficiency was about 78.2 ± 6.3% (mean ± SD), which was considered to be adequate for the functional analyses.
TextSentencer_T207 30281-30408 Sentence denotes The transfection efficiency was about 78.2 ± 6.3% (mean ± SD), which was considered to be adequate for the functional analyses.
TextSentencer_T208 30409-30467 Sentence denotes The human miR-1 miRNA was also used as a positive control.
TextSentencer_T208 30409-30467 Sentence denotes The human miR-1 miRNA was also used as a positive control.
TextSentencer_T209 30468-30712 Sentence denotes In this control, the human miR-1 miRNA mimic effectively down-regulated the expression of twinfilin-1 (also known as PTK9) by 80% at the mRNA level as detected by real-time PCR using TaqMan® Gene Expression Assays (Applied Biosystems) for PTK9.
TextSentencer_T209 30468-30712 Sentence denotes In this control, the human miR-1 miRNA mimic effectively down-regulated the expression of twinfilin-1 (also known as PTK9) by 80% at the mRNA level as detected by real-time PCR using TaqMan® Gene Expression Assays (Applied Biosystems) for PTK9.
TextSentencer_T210 30713-30805 Sentence denotes This positive control proved the effective delivery and activity of Pre-miR miRNA Precursor.
TextSentencer_T210 30713-30805 Sentence denotes This positive control proved the effective delivery and activity of Pre-miR miRNA Precursor.
TextSentencer_T211 30806-30869 Sentence denotes We therefore used this model in further functional experiments.
TextSentencer_T211 30806-30869 Sentence denotes We therefore used this model in further functional experiments.
TextSentencer_T212 30870-30914 Sentence denotes Each experiment was performed in triplicate.
TextSentencer_T212 30870-30914 Sentence denotes Each experiment was performed in triplicate.