Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference g...Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference gene and target gene would be degraded.As a result,it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation.This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment.Methods The new RNA was labeled with 5-ethyluridine(5-EU)instead of uracil,and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of“Click Chemistry”and magnetic bead screening.Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon(M.B.)and quantitative reverse transcription PCR(qRT-PCR).Results The bacterial nascent RNA captured by“Click Chemistry”screening can be used as a reverse transcription template to form cDNA.Combined with the fluorescent molecular beacon M.B.1,the synthesis rate of rRNA at 37℃is 1.2 times higher than that at 15℃.The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃and 16℃when analyzed with nascent RNA rather than total RNA,enabling accurate detection of RNA transcription rates.Conclusion Compared to other article reported experimental methods that utilize screening magnetic columns,the technical scheme employed in this study is more suitable for bacteria,and the operation steps are simple and easy to implement,making it an effective RNA capture method for researchers.展开更多
For the low-grade copper sulfide ores with 0.99% of copper, of which 41.5% was primary copper sulfide, and 54.5% was secondary copper sulfide, well-controlled column bioleaching on a novel equipment was carried out to...For the low-grade copper sulfide ores with 0.99% of copper, of which 41.5% was primary copper sulfide, and 54.5% was secondary copper sulfide, well-controlled column bioleaching on a novel equipment was carried out to investigate the optimal conditions of pre-leaching, particle sizes of ores, temperature, spray intensity and strain consortium. Results show that copper extraction of 91.11% can be obtained after 90 d with the optimal p H value of pre-leaching of 0.8; the p H values of pre-leaching significantly affect the final copper extractions. Copper extractions of 93.11%, 91.04% and 80.45% can be obtained for the bioleaching of ores with particles size of 5-8 mm, 5-15 mm and 5-20 mm, respectively. Copper extractions are 83.77% and 91.02% for bioleaching under the conditions of room temperature and 35 oC. Copper extractions are 77.25%, 85.45% and 91.12% for the bioleaching when flow rate of spray was 5 L/（h·m2）, 10 L/（h·m2） and 15 L/（h·m2）, respectively. Additionally, the strain consortium C3 is the best among the four strain consortia in bioleaching. By considering the energy consumption, the optimal conditions of bioleaching in this work are determined as p H of pre-leaching of 0.8, particles size of 5-15 mm, temperature of 35 ℃, spray intensity of 15 L/（h·m2）, and strain consortium C3.展开更多
To enhance the nitrogen removal,a systemic monitoring of the biological and hydrological parameters of Carrousel oxidation ditch in Chongqing Jingkou Wastewater Treatment Plant was carried out to study the feasibility...To enhance the nitrogen removal,a systemic monitoring of the biological and hydrological parameters of Carrousel oxidation ditch in Chongqing Jingkou Wastewater Treatment Plant was carried out to study the feasibility of simultaneous nitrification and denitrification(SND).The variation and distribution of parameters such as flow velocity,concentration of dissolved oxygen(DO) and mixed liquor suspended solids(MLSS) in oxidation ditch were monitored and analyzed,which were major control factors for SND.The results showed that,the dimensional distribution of flow velocity,DO and MLSS were affected significantly by the operation condition of the aeration wheels.With all the four aeration wheels being in operation,DO and flow velocity were higher and the mixing of MLSS was sufficient.With three aeration wheels being in operation,the flow velocity in most of the bottom areas was enough to meet the basic requirements of no deposition,and the anaerobic region and aerobic region could exist simultaneously in one oxidation ditch,which was helpful to the process of SND.According to spatial distribution characteristics of the flow velocity,DO and soluble components under optimized condition,different functional zones of biochemical reaction in the Carrousel oxidation ditch system were defined,which might contribute to the optimization control and SND of Carrousel oxidation ditch.展开更多
文摘Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference gene and target gene would be degraded.As a result,it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation.This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment.Methods The new RNA was labeled with 5-ethyluridine(5-EU)instead of uracil,and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of“Click Chemistry”and magnetic bead screening.Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon(M.B.)and quantitative reverse transcription PCR(qRT-PCR).Results The bacterial nascent RNA captured by“Click Chemistry”screening can be used as a reverse transcription template to form cDNA.Combined with the fluorescent molecular beacon M.B.1,the synthesis rate of rRNA at 37℃is 1.2 times higher than that at 15℃.The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃and 16℃when analyzed with nascent RNA rather than total RNA,enabling accurate detection of RNA transcription rates.Conclusion Compared to other article reported experimental methods that utilize screening magnetic columns,the technical scheme employed in this study is more suitable for bacteria,and the operation steps are simple and easy to implement,making it an effective RNA capture method for researchers.
基金Projects(51374248,51320105006) supported by National Natural Science Foundation of ChinaProject(NCET-13-0595) supported by the Program for New Century Excellent Talents in University,ChinaProject(2014T70692) supported by the China Postdoctoral Science Foundation
文摘For the low-grade copper sulfide ores with 0.99% of copper, of which 41.5% was primary copper sulfide, and 54.5% was secondary copper sulfide, well-controlled column bioleaching on a novel equipment was carried out to investigate the optimal conditions of pre-leaching, particle sizes of ores, temperature, spray intensity and strain consortium. Results show that copper extraction of 91.11% can be obtained after 90 d with the optimal p H value of pre-leaching of 0.8; the p H values of pre-leaching significantly affect the final copper extractions. Copper extractions of 93.11%, 91.04% and 80.45% can be obtained for the bioleaching of ores with particles size of 5-8 mm, 5-15 mm and 5-20 mm, respectively. Copper extractions are 83.77% and 91.02% for bioleaching under the conditions of room temperature and 35 oC. Copper extractions are 77.25%, 85.45% and 91.12% for the bioleaching when flow rate of spray was 5 L/（h·m2）, 10 L/（h·m2） and 15 L/（h·m2）, respectively. Additionally, the strain consortium C3 is the best among the four strain consortia in bioleaching. By considering the energy consumption, the optimal conditions of bioleaching in this work are determined as p H of pre-leaching of 0.8, particles size of 5-15 mm, temperature of 35 ℃, spray intensity of 15 L/（h·m2）, and strain consortium C3.
基金Project(2009ZX07315-002-01) supported by the Water Pollution Control and Management of Major Special Science and Technology, China Project(CDJXS11210001) supported by the Scientific and Technical Innovation Project of Chongqing University Graduation Foundation, China
文摘To enhance the nitrogen removal,a systemic monitoring of the biological and hydrological parameters of Carrousel oxidation ditch in Chongqing Jingkou Wastewater Treatment Plant was carried out to study the feasibility of simultaneous nitrification and denitrification(SND).The variation and distribution of parameters such as flow velocity,concentration of dissolved oxygen(DO) and mixed liquor suspended solids(MLSS) in oxidation ditch were monitored and analyzed,which were major control factors for SND.The results showed that,the dimensional distribution of flow velocity,DO and MLSS were affected significantly by the operation condition of the aeration wheels.With all the four aeration wheels being in operation,DO and flow velocity were higher and the mixing of MLSS was sufficient.With three aeration wheels being in operation,the flow velocity in most of the bottom areas was enough to meet the basic requirements of no deposition,and the anaerobic region and aerobic region could exist simultaneously in one oxidation ditch,which was helpful to the process of SND.According to spatial distribution characteristics of the flow velocity,DO and soluble components under optimized condition,different functional zones of biochemical reaction in the Carrousel oxidation ditch system were defined,which might contribute to the optimization control and SND of Carrousel oxidation ditch.
