AIM: To test whether the replication of human rhinovirus(HRV) is regulated by micro RNAs in human bronchial epithelial cells.METHODS: For the present study, the human cell line BEAS-2B(derived from normal human bronch...AIM: To test whether the replication of human rhinovirus(HRV) is regulated by micro RNAs in human bronchial epithelial cells.METHODS: For the present study, the human cell line BEAS-2B(derived from normal human bronchial epithelial cells) was adopted. DICER knock-down, by si RNA transfection in BEAS-2B cells, was performed in order to inhibit micro RNA maturation globally. Alternatively, antisense oligonucleotides(anti-mi Rs) were transfectedto inhibit the activity of specific micro RNAs. Cells were infected with HRV-1B. Viral replication was assessed by measuring the genomic viral RNA by reverse transcription quantitative polymerase chain reaction(RT-q PCR). Association between micro RNA-induced-silencing-complex and viral RNA was detected by Ago2 co-immunoprecipitation followed by RT-q PCR. Targetscan v.6 was used to predict micro RNA target sites on several HRV strains.RESULTS: Here, we show that micro RNAs affect replication of HRV-1B. DICER knock-down significantly reduced the expression of mature micro RNAs in a bronchial epithelial cell line(BEAS-2B) and in turn, increased the synthesis of HRV-1B RNA. Additionally, HRV-1B RNA co-immunoprecipitated with argonaute 2 protein, an important effector for micro RNA activity suggesting that micro RNAs bind to viral RNA during infection. In order to identify specific micro RNAs involved in this interaction, we employed bioinformatics analysis, and selected a group of micro RNAs that have been reported to be under-expressed in asthmatic bronchial epithelial cells and were predicted to target different strains of rhinoviruses(HRV-1B,-16,-14,-27). Our results suggest that, out of this group of micro RNAs, mi R-128 and mi R-155 contribute to the innate defense against HRV-1B: transfection of specific anti-mi Rs increased viral replication, as anticipated in-silico.CONCLUSION: Taken together, our results suggest that pathological changes in micro RNA expression, as already reported for asthma or chronic obstructive pulmonary disease have the potential to affect Rhinovirus replication and therefore may play a role in virusinduced exacerbations.展开更多
Recent studies demonstrate that many avialan features evolved incrementally prior to the origin of the group,but the presence of some of these features,such as bird-like brooding behaviours,remains contentious in non-...Recent studies demonstrate that many avialan features evolved incrementally prior to the origin of the group,but the presence of some of these features,such as bird-like brooding behaviours,remains contentious in non-avialan dinosaurs.Here we report the first non-avialan dinosaur fossil known to preserve an adult skeleton atop an egg clutch that contains embryonic remains.The preserved positional relationship of the adult to the clutch,coupled with the advanced growth stages of the embryos and their high estimated incubation temperatures,provides strong support for the brooding hypothesis.Furthermore,embryos in the clutch are at different developmental stages,suggesting the presence of asynchronous hatching—a derived feature even among crown-group birds—in non-avialan theropods.These findings demonstrate that the evolution of reproductive biology along bird-line archosaurs was a complex rather than a linear and incremental process,and suggest that some aspects of non-avialan theropod reproduction were unique to these dinosaurs.展开更多
基金Supported by MRC,AAIR and the Roger Brooke charitable trust
文摘AIM: To test whether the replication of human rhinovirus(HRV) is regulated by micro RNAs in human bronchial epithelial cells.METHODS: For the present study, the human cell line BEAS-2B(derived from normal human bronchial epithelial cells) was adopted. DICER knock-down, by si RNA transfection in BEAS-2B cells, was performed in order to inhibit micro RNA maturation globally. Alternatively, antisense oligonucleotides(anti-mi Rs) were transfectedto inhibit the activity of specific micro RNAs. Cells were infected with HRV-1B. Viral replication was assessed by measuring the genomic viral RNA by reverse transcription quantitative polymerase chain reaction(RT-q PCR). Association between micro RNA-induced-silencing-complex and viral RNA was detected by Ago2 co-immunoprecipitation followed by RT-q PCR. Targetscan v.6 was used to predict micro RNA target sites on several HRV strains.RESULTS: Here, we show that micro RNAs affect replication of HRV-1B. DICER knock-down significantly reduced the expression of mature micro RNAs in a bronchial epithelial cell line(BEAS-2B) and in turn, increased the synthesis of HRV-1B RNA. Additionally, HRV-1B RNA co-immunoprecipitated with argonaute 2 protein, an important effector for micro RNA activity suggesting that micro RNAs bind to viral RNA during infection. In order to identify specific micro RNAs involved in this interaction, we employed bioinformatics analysis, and selected a group of micro RNAs that have been reported to be under-expressed in asthmatic bronchial epithelial cells and were predicted to target different strains of rhinoviruses(HRV-1B,-16,-14,-27). Our results suggest that, out of this group of micro RNAs, mi R-128 and mi R-155 contribute to the innate defense against HRV-1B: transfection of specific anti-mi Rs increased viral replication, as anticipated in-silico.CONCLUSION: Taken together, our results suggest that pathological changes in micro RNA expression, as already reported for asthma or chronic obstructive pulmonary disease have the potential to affect Rhinovirus replication and therefore may play a role in virusinduced exacerbations.
基金supported by the Double First-Class Joint Program of Yunnan Science and Technology Department and Yunnan University (2018FY001-005)the China-Myanmar Joint Laboratory for Ecological and Environmental Conservation+2 种基金the University of Hong Kong Faculty of Science RAE Improvement Fundsupported by the CNRS Program INSU INTERRVIEthe National Natural Science Foundation of China(41688103)。
文摘Recent studies demonstrate that many avialan features evolved incrementally prior to the origin of the group,but the presence of some of these features,such as bird-like brooding behaviours,remains contentious in non-avialan dinosaurs.Here we report the first non-avialan dinosaur fossil known to preserve an adult skeleton atop an egg clutch that contains embryonic remains.The preserved positional relationship of the adult to the clutch,coupled with the advanced growth stages of the embryos and their high estimated incubation temperatures,provides strong support for the brooding hypothesis.Furthermore,embryos in the clutch are at different developmental stages,suggesting the presence of asynchronous hatching—a derived feature even among crown-group birds—in non-avialan theropods.These findings demonstrate that the evolution of reproductive biology along bird-line archosaurs was a complex rather than a linear and incremental process,and suggest that some aspects of non-avialan theropod reproduction were unique to these dinosaurs.
