A型牛轮状病毒(BRV-A)是引起牛腹泻的重要病原之一,对养牛业危害最大。为建立一种针对BRV-A的快速、高通量、低成本检测方法,根据BRV-A的VP6基因保守区设计特异性引物,建立了检测BRV-A的SYBR Green I RT-qPCR方法。该方法对浓度梯度为4....A型牛轮状病毒(BRV-A)是引起牛腹泻的重要病原之一,对养牛业危害最大。为建立一种针对BRV-A的快速、高通量、低成本检测方法,根据BRV-A的VP6基因保守区设计特异性引物,建立了检测BRV-A的SYBR Green I RT-qPCR方法。该方法对浓度梯度为4.41×10^(8)~4.41×10^(2) copies/μL质粒标准品的扩增Ct值与拷贝数浓度呈良好的线性关系,熔解曲线为单峰;与其他引起牛腹泻的常见病原无交叉反应,最低检测限为4.41×10^(1) copies/μL,批内、批间重复性试验Ct值变异系数均低于1%;对临床粪便样品的阳性检出率高于地方标准中的PCR方法,且检测结果符合性较好。综上,本研究建立的BRV-A SYBR Green I RT-qPCR检测方法灵敏、特异、稳定,且操作简单、成本低,为临床样品的大规模BRV-A检测及其感染的早期诊断提供了技术支撑。展开更多
为建立牛副流感病毒5型(BPIV5)的SYBR Green Ⅰ荧光定量PCR(qPCR)检测方法,本研究经PCR扩增BPIV5 L基因的保守片段,构建重组质粒p MD18-T-BPIV5,并经PCR、双酶切与测序鉴定正确后作为标准品,经各反应条件优化后初步建立检测BPIV5的qPCR...为建立牛副流感病毒5型(BPIV5)的SYBR Green Ⅰ荧光定量PCR(qPCR)检测方法,本研究经PCR扩增BPIV5 L基因的保守片段,构建重组质粒p MD18-T-BPIV5,并经PCR、双酶切与测序鉴定正确后作为标准品,经各反应条件优化后初步建立检测BPIV5的qPCR方法。利用10倍倍比稀释的质粒标准品作为模板,利用优化的qPCR扩增,建立该方法的标准曲线,结果显示,重组质粒标准品的拷贝数与其Ct值呈良好的线性关系,相关系数为0.998,扩增效率为1.09。以BPIV5、牛肠道病毒(BEV)、牛病毒性腹泻病毒(BVDV)、牛诺如病毒(BNOV)、牛副流感病毒3型(BPIV3)和牛冠状病毒(BCOV)的cDNA作为模板,采用该qPCR扩增,评估该方法的特异性;以10倍倍比稀释的重组质粒标准品pMD18-T-BPIV5作为模板,利用本研究建立的qPCR方法扩增,评估该方法的敏感性;以3种不同浓度的质粒标准品作为模板,利用本研究建立的qPCR方法在同一时间和不同时间分别进行批内和批间重复性试验。结果显示,该方法仅能特异性扩增BPIV5,其他相关病毒均为阴性结果;对质粒标准品的检测限为6.43拷贝/μL;批内和批间重复性试验的变异系数均小于2%。利用该方法和已发表的qPCR方法分别检测采自内蒙古东部部分牛场的80份鼻拭子样品,结果显示,两种方法对样品的阳性检测率均为10%(8/80),阴性样品的检测率均为90%(72/80),二者的阳性符合率和总符合率均达100%。本研究建立的检测BPIV5的qPCR方法特异性较强、敏感性较高、重复性较好,可以用于临床样品的检测,为BPIV5的检测和流行病学调查提供了一种新的检测手段。展开更多
In the era of green logistics,digital transformation has become an effective means for the logistics industry’s high-quality development.Using listed companies in China’s logistics industry from 2010 to 2021 as the ...In the era of green logistics,digital transformation has become an effective means for the logistics industry’s high-quality development.Using listed companies in China’s logistics industry from 2010 to 2021 as the research samples,this paper conducts an empirical test on the impact of the digital transformation of logistics enterprises on their green in-novation.Specifically,enterprise digital transformation indicators are constructed through the text analysis method,and the fixed-effects model is applied for analysis.The results indicate that the digital transformation of logistics enterprises has a significant promoting effect on their green innovation;the promoting effect of the digital transformation of logistics enterprises on green innovation is primarily achieved by easing corporate financing constraints and reducing corporate en-vironmental uncertainty;and the impact of digital transformation on green innovation is geographically heterogeneous.展开更多
A strategy for the green synthesis of heterocyclicβ-ketosulfides via nucleophilic substitution ofα-halogenated ketone with het-eroaryl thiols in water media is presented.Compared with the available literature report...A strategy for the green synthesis of heterocyclicβ-ketosulfides via nucleophilic substitution ofα-halogenated ketone with het-eroaryl thiols in water media is presented.Compared with the available literature reports,this new method had the advantages of base-free,additives-free,simple operation,mild condition,greenness,high efficiency,tolerance of a broad scope of substrates.Furth-more,the reaction could easily be scaled up in gram scale and the products also could easily transformed to other useful organic compounds.Mechanism investigation indicated that the tautomerism of pyrimidine-2-thiol to pyrimidine-2(1H)-thione and the hy-drogen bonds played important roles in the reaction.展开更多
Novel green ceramic pigments Y_(3)Ga_(3)MgSiO_(12)∶xCr^(3+)(x=0−0.2)were successfully synthesized via the conventional solid-state approach.The properties of the pigments were studied by XRD,FE-SEM,UV-Vis spectroscop...Novel green ceramic pigments Y_(3)Ga_(3)MgSiO_(12)∶xCr^(3+)(x=0−0.2)were successfully synthesized via the conventional solid-state approach.The properties of the pigments were studied by XRD,FE-SEM,UV-Vis spectroscopy,XPS,and chromaticity analysis.The findings reveal that the trivalent chromium ions occupy the[Ga1O6]octahedral sites within the garnet lattice,and the relatively weak crystal field environment provided by the matrix endows the pigments with green characteristics.The samples prepared by calcination at 1400℃exhibit the most excellent performance in terms of phase purity,morphology,and color properties.The chromaticity values for the representative sample Y_(3)Ga_(3)MgSiO_(12)∶0.05Cr^(3+)are L^(*)=81.16,a^(*)=−12.53,and b^(*)=12.71,and the color remains stable after the stability test.Moreover,when glazed with Y_(3)Ga_(3)MgSiO_(12)∶xCr^(3+)(x=0−0.2)pigments,the smooth glaze surfaces exhibit vivid and saturated green tones,demonstrating their remarkable coloring capabilities and promising potential as a practical pigment for medium-temperature applications.This research underscores the vast application prospects of Y_(3)Ga_(3)MgSiO_(12)∶Cr^(3+)as an innovative green ceramic pigment.展开更多
Herein,a one-pot chemical reduction method was reported to prepare folic acid(FA)-stabilized silver nanoclusters(FA@Ag NCs),in which FA,hydrazine hydrate,and silver nitrate were used as capping agent,reducing agent,an...Herein,a one-pot chemical reduction method was reported to prepare folic acid(FA)-stabilized silver nanoclusters(FA@Ag NCs),in which FA,hydrazine hydrate,and silver nitrate were used as capping agent,reducing agent,and precursor,respectively.Several technologies were employed to investigate the structures and optical properties of FA@Ag NCs,including transmission electron microscopy(TEM),X-ray photoelectron spectrometer(XPS),Fourier transform infrared spectrometer(FTIR),X-ray diffractometer(XRD),fluorescence spectrometer,and ultraviolet visible absorption spectrometer.FA@Ag NCs were suggested to be highly dispersed and spherical with a size of around 2.8 nm.Moreover,the maximum excitation and emission wavelengths of FA@Ag NCs were 370 and 447 nm,respectively.Under the optimal detection conditions,FA@Ag NCs could be used to effectively detect malachite green with the linear detection range of 0.5-200μmol·L^(-1).The detection limit was 0.084μmol·L^(-1).The fluorescence-quenching mechanism was ascribed to the static quenching.The detection system based on FA@AgNCs was successfully used for the detection of malachite green in actual samples with good accuracy and reproducibility.展开更多
The National Independent Innovation Demonstration Zone has been assigned the unique mission of demonstrating and leading national innovation and playing a key supportive role in enhancing green innovation.Based on the...The National Independent Innovation Demonstration Zone has been assigned the unique mission of demonstrating and leading national innovation and playing a key supportive role in enhancing green innovation.Based on the sample data of A-share listed companies in China from 2007 to 2021,we apply a multi-period difference-in-differences model to analyze whether the implementation of the National Independent Innovation Demonstration Zone policies plays a leverage effect or a crowd out effect on the green innovation efficiency of enterprises and systematically test the regulatory mechanism of government grants and media attention in the process of this influence.The empirical results show that the imple-mentation of the National Independent Innovation Demonstration Zone policies has a positive impact on the green innovation efficiency of enterprises and that the green innovation induced by this reform is not the leverage effect of additional R&D investment on the basis of the existing innovation activities of enterprises but rather the result of the reallocation of resources to crowd out existing non-green innovation.It is further found that government grants and media attention positively moderate the positive driving effect of National Independent Innovation Demonstration Zone policies on the green innovation efficiency of enterprises.展开更多
为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38....为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38.77;除WPV出现S形扩增曲线外,新城疫病毒(NDV)、H9亚型禽流感病毒(H9 AIV)、鸭坦布苏病毒(DTMUV)、鸭肝炎病毒(DHAV)、鸭肠炎病毒(DEV)、鸭呼肠孤病毒(DRV)样品均未出现S形阳性扩增曲线;批内变异系数(CV)为0.15%~0.23%,批间变异系数为0.09%~0.28%。结果表明,SYBR Green Ⅰ荧光定量PCR检测方法重复性好、灵敏度高和特异性强。临床样品检测结果表明,SYBR Green Ⅰ荧光定量PCR与普通PCR的符合率达98.4%,灵敏度是普通PCR的1 000倍。SYBR Green Ⅰ荧光定量PCR检测方法不仅能定性检测WPV,还可以进行定量检测,可用于种鸭场、种鹅场的WPV净化检测,也可用于WPV临床大量样品的快速检测。展开更多
为了建立高效、灵敏的猪流行性腹泻病毒(PEDV)检测方法,本研究从GenBank数据库中获取PEDV N基因序列,扩增出PEDV N基因标准质粒,并在N基因的保守区域内设计了一对特异性荧光定量引物,成功建立了SYBR Green I实时荧光定量PCR检测方法。...为了建立高效、灵敏的猪流行性腹泻病毒(PEDV)检测方法,本研究从GenBank数据库中获取PEDV N基因序列,扩增出PEDV N基因标准质粒,并在N基因的保守区域内设计了一对特异性荧光定量引物,成功建立了SYBR Green I实时荧光定量PCR检测方法。经过一系列试验表明,该检测方法线性关系良好,R^(2)值为0.99;特异性强,敏感性高,最低可检测至2.23 copies/μL,比普通PCR灵敏约100倍;重复性好,组内变异系数为0.25%~0.43%,组间变异系数为0.67%~0.97%;对于各地区96份临床样品检测出PEDV阳性率为25%。本研究建立的实时荧光定量PCR检测方法为PEDV的临床诊断、流行病学调查以及定量研究提供了有效的检测工具。展开更多
文摘A型牛轮状病毒(BRV-A)是引起牛腹泻的重要病原之一,对养牛业危害最大。为建立一种针对BRV-A的快速、高通量、低成本检测方法,根据BRV-A的VP6基因保守区设计特异性引物,建立了检测BRV-A的SYBR Green I RT-qPCR方法。该方法对浓度梯度为4.41×10^(8)~4.41×10^(2) copies/μL质粒标准品的扩增Ct值与拷贝数浓度呈良好的线性关系,熔解曲线为单峰;与其他引起牛腹泻的常见病原无交叉反应,最低检测限为4.41×10^(1) copies/μL,批内、批间重复性试验Ct值变异系数均低于1%;对临床粪便样品的阳性检出率高于地方标准中的PCR方法,且检测结果符合性较好。综上,本研究建立的BRV-A SYBR Green I RT-qPCR检测方法灵敏、特异、稳定,且操作简单、成本低,为临床样品的大规模BRV-A检测及其感染的早期诊断提供了技术支撑。
文摘为建立牛副流感病毒5型(BPIV5)的SYBR Green Ⅰ荧光定量PCR(qPCR)检测方法,本研究经PCR扩增BPIV5 L基因的保守片段,构建重组质粒p MD18-T-BPIV5,并经PCR、双酶切与测序鉴定正确后作为标准品,经各反应条件优化后初步建立检测BPIV5的qPCR方法。利用10倍倍比稀释的质粒标准品作为模板,利用优化的qPCR扩增,建立该方法的标准曲线,结果显示,重组质粒标准品的拷贝数与其Ct值呈良好的线性关系,相关系数为0.998,扩增效率为1.09。以BPIV5、牛肠道病毒(BEV)、牛病毒性腹泻病毒(BVDV)、牛诺如病毒(BNOV)、牛副流感病毒3型(BPIV3)和牛冠状病毒(BCOV)的cDNA作为模板,采用该qPCR扩增,评估该方法的特异性;以10倍倍比稀释的重组质粒标准品pMD18-T-BPIV5作为模板,利用本研究建立的qPCR方法扩增,评估该方法的敏感性;以3种不同浓度的质粒标准品作为模板,利用本研究建立的qPCR方法在同一时间和不同时间分别进行批内和批间重复性试验。结果显示,该方法仅能特异性扩增BPIV5,其他相关病毒均为阴性结果;对质粒标准品的检测限为6.43拷贝/μL;批内和批间重复性试验的变异系数均小于2%。利用该方法和已发表的qPCR方法分别检测采自内蒙古东部部分牛场的80份鼻拭子样品,结果显示,两种方法对样品的阳性检测率均为10%(8/80),阴性样品的检测率均为90%(72/80),二者的阳性符合率和总符合率均达100%。本研究建立的检测BPIV5的qPCR方法特异性较强、敏感性较高、重复性较好,可以用于临床样品的检测,为BPIV5的检测和流行病学调查提供了一种新的检测手段。
基金supported by the National Natural Science Foundation of China(72374061,72204243)the Ministry of Education’s Humanities and Social Science Research Youth Fund Project(20YJC630138,22YJC630056)+1 种基金Anhui Provincial Natural Science Foundation(2208085UD02)New Liberal Arts Fund Expansion Project of University of Science and Technology of China(FSSF-A-230317).
文摘In the era of green logistics,digital transformation has become an effective means for the logistics industry’s high-quality development.Using listed companies in China’s logistics industry from 2010 to 2021 as the research samples,this paper conducts an empirical test on the impact of the digital transformation of logistics enterprises on their green in-novation.Specifically,enterprise digital transformation indicators are constructed through the text analysis method,and the fixed-effects model is applied for analysis.The results indicate that the digital transformation of logistics enterprises has a significant promoting effect on their green innovation;the promoting effect of the digital transformation of logistics enterprises on green innovation is primarily achieved by easing corporate financing constraints and reducing corporate en-vironmental uncertainty;and the impact of digital transformation on green innovation is geographically heterogeneous.
文摘A strategy for the green synthesis of heterocyclicβ-ketosulfides via nucleophilic substitution ofα-halogenated ketone with het-eroaryl thiols in water media is presented.Compared with the available literature reports,this new method had the advantages of base-free,additives-free,simple operation,mild condition,greenness,high efficiency,tolerance of a broad scope of substrates.Furth-more,the reaction could easily be scaled up in gram scale and the products also could easily transformed to other useful organic compounds.Mechanism investigation indicated that the tautomerism of pyrimidine-2-thiol to pyrimidine-2(1H)-thione and the hy-drogen bonds played important roles in the reaction.
文摘Novel green ceramic pigments Y_(3)Ga_(3)MgSiO_(12)∶xCr^(3+)(x=0−0.2)were successfully synthesized via the conventional solid-state approach.The properties of the pigments were studied by XRD,FE-SEM,UV-Vis spectroscopy,XPS,and chromaticity analysis.The findings reveal that the trivalent chromium ions occupy the[Ga1O6]octahedral sites within the garnet lattice,and the relatively weak crystal field environment provided by the matrix endows the pigments with green characteristics.The samples prepared by calcination at 1400℃exhibit the most excellent performance in terms of phase purity,morphology,and color properties.The chromaticity values for the representative sample Y_(3)Ga_(3)MgSiO_(12)∶0.05Cr^(3+)are L^(*)=81.16,a^(*)=−12.53,and b^(*)=12.71,and the color remains stable after the stability test.Moreover,when glazed with Y_(3)Ga_(3)MgSiO_(12)∶xCr^(3+)(x=0−0.2)pigments,the smooth glaze surfaces exhibit vivid and saturated green tones,demonstrating their remarkable coloring capabilities and promising potential as a practical pigment for medium-temperature applications.This research underscores the vast application prospects of Y_(3)Ga_(3)MgSiO_(12)∶Cr^(3+)as an innovative green ceramic pigment.
文摘Herein,a one-pot chemical reduction method was reported to prepare folic acid(FA)-stabilized silver nanoclusters(FA@Ag NCs),in which FA,hydrazine hydrate,and silver nitrate were used as capping agent,reducing agent,and precursor,respectively.Several technologies were employed to investigate the structures and optical properties of FA@Ag NCs,including transmission electron microscopy(TEM),X-ray photoelectron spectrometer(XPS),Fourier transform infrared spectrometer(FTIR),X-ray diffractometer(XRD),fluorescence spectrometer,and ultraviolet visible absorption spectrometer.FA@Ag NCs were suggested to be highly dispersed and spherical with a size of around 2.8 nm.Moreover,the maximum excitation and emission wavelengths of FA@Ag NCs were 370 and 447 nm,respectively.Under the optimal detection conditions,FA@Ag NCs could be used to effectively detect malachite green with the linear detection range of 0.5-200μmol·L^(-1).The detection limit was 0.084μmol·L^(-1).The fluorescence-quenching mechanism was ascribed to the static quenching.The detection system based on FA@AgNCs was successfully used for the detection of malachite green in actual samples with good accuracy and reproducibility.
基金supported by the National Natural Science Foundation of China(72474034)the Young Elite Scientists Sponsorship Program by SAST(20240123)+1 种基金Humanities and Social Science Fund of Ministry of Education of China(21YJC630037,21YJC630057)Social Science Foundation of Xi’an(25JX218).
文摘The National Independent Innovation Demonstration Zone has been assigned the unique mission of demonstrating and leading national innovation and playing a key supportive role in enhancing green innovation.Based on the sample data of A-share listed companies in China from 2007 to 2021,we apply a multi-period difference-in-differences model to analyze whether the implementation of the National Independent Innovation Demonstration Zone policies plays a leverage effect or a crowd out effect on the green innovation efficiency of enterprises and systematically test the regulatory mechanism of government grants and media attention in the process of this influence.The empirical results show that the imple-mentation of the National Independent Innovation Demonstration Zone policies has a positive impact on the green innovation efficiency of enterprises and that the green innovation induced by this reform is not the leverage effect of additional R&D investment on the basis of the existing innovation activities of enterprises but rather the result of the reallocation of resources to crowd out existing non-green innovation.It is further found that government grants and media attention positively moderate the positive driving effect of National Independent Innovation Demonstration Zone policies on the green innovation efficiency of enterprises.
文摘为了建立水禽细小病毒(WPV)快速检测方法,根据序列比对结果在水禽细小病毒NS基因SF3保守区域内设计特异性引物,建立SYBR Green Ⅰ荧光定量PCR通用检测方法。该方法的扩增效率(E)为90.0%,相关系数(R~2)=0.99,标准曲线方程为y=-3.607x+38.77;除WPV出现S形扩增曲线外,新城疫病毒(NDV)、H9亚型禽流感病毒(H9 AIV)、鸭坦布苏病毒(DTMUV)、鸭肝炎病毒(DHAV)、鸭肠炎病毒(DEV)、鸭呼肠孤病毒(DRV)样品均未出现S形阳性扩增曲线;批内变异系数(CV)为0.15%~0.23%,批间变异系数为0.09%~0.28%。结果表明,SYBR Green Ⅰ荧光定量PCR检测方法重复性好、灵敏度高和特异性强。临床样品检测结果表明,SYBR Green Ⅰ荧光定量PCR与普通PCR的符合率达98.4%,灵敏度是普通PCR的1 000倍。SYBR Green Ⅰ荧光定量PCR检测方法不仅能定性检测WPV,还可以进行定量检测,可用于种鸭场、种鹅场的WPV净化检测,也可用于WPV临床大量样品的快速检测。
