水禽细小病毒SYBR Green Ⅰ荧光定量PCR检测方法的建立与应用

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。

盆底超声检查和Green分型诊断产妇产后早期盆底结构及压力性尿失禁

目的:探究盆底超声检查和Green分型在产妇产后早期盆底结构及压力性尿失禁(SUI)诊断中的价值。方法:选取2021年3月-2023年3月本院就诊经产妇204例,根据产后早期(6~8周)SUI发生情况分为SUI组(82例)和非SUI组(122例)。检查产妇静息和Valsalva动作状态下膀胱颈距耻骨联合下缘的距离(BSD)、膀胱尿道后角(PUA)、膀胱颈移动度(BND)和尿道旋转角(UR),对比Green分型;采用受试者工作特征(ROC)曲线分析相关参数诊断SUI价值。结果:静息状态下,两组盆底超声参数无差异(P>0.05);SUI组Valsalva动作状态下的BSD、PUA、BND和UR值均大于非SUI组,Green分型Ι型占比(25.6%)低于非SUI组(59.8%)(均P<0.05)。ROC曲线分析,Green分型曲线下面积(AUC)0.579,对SUI诊断价值不佳(P=0.057),Valsalva动作状态下BSD(AUC=0.758)、PUA(AUC=0.743)、BND(AUC=0.769)和UR(AUC=0.705)对SUI诊断价值良好(均P<0.05)。结论:盆底超声检查和Green分型能有效评估经产妇早期盆底结构状况,前者可诊断SUI。

约束Green函数与非线性地基梁模态分析

在生物和医学领域,微机电系统(MEMS)中的微梁结构在植入人体的使用时,由于体内的细胞环境类似于水凝胶,在这种环境下工作,设备和仪器的精度和稳定性很大程度上受到细胞弹性的影响.为了分析此类地基梁的动力学问题,本文建立了非线性基础上的梁振动模型,研究了任意位置弹簧和非线性弹簧基础上的梁模态.通过Laplace变换和线性叠加原理,得到了一种约束Green函数,利用数值计算验证方案的有效性,并研究了各种重要物理参数的影响,发现弹簧位置向跨中移动时,模态对称性被打破,弹簧刚度增加,模态阶数改变.

猪流行性腹泻病毒SYBR Green I实时荧光定量PCR检测方法的建立

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。

美洲鳗鲡腺瘤病毒(AEAdoV)普通PCR和SYBR Green I实时荧光定量PCR检测方法的建立及应用

为建立美洲鳗鲡腺瘤病毒(AEAdoV)的检测方法,根据AEAdoV福建株(AEAdoVFJ)的superfamily 3 helicases(S3H)序列,设计引物,建立了AEAdoV的普通PCR和qPCR检测方法;进一步评价检测方法的灵敏性、特异性及重复性,利用2种方法对美洲鳗鲡“出血性烂鳃”病料进行了检测,并对美洲鳗鲡体内不同组织的病毒含量进行分析。结果显示,普通PCR扩增的目的片段长度约300 bp,利用其构建的qPCR质粒标准品,其拷贝数与qPCR阈值循环数(C_(t))线性关系良好,线性范围广,标准曲线相关系数(R2)达到0.999,扩增效率为105.067%。建立的普通PCR法和qPCR的最低检测AEAdoV拷贝数分别为100个和10个。2种方法均可特异性检测AEAdoV,而对蛙虹彩病毒(RGV)、鳗鲡疱疹病毒(AngHV)、鲤疱疹病毒(KHV)、对虾白斑综合征病毒(WSSV)、日本鳗鲡内皮细胞坏死病毒(JEAdoV)和花鳗鲡腺瘤病毒(MEAdoV)均无扩增反应。qPCR法的组内和组间变异系数均小于2%,表明其重复性良好。临床应用结果显示,35份美洲鳗鲡“出血性烂鳃”病料,采用普通PCR法的AEAdoV检出率为82.8%,而qPCR法的AEAdoV检出率为97%。对美洲鳗鲡不同组织的病毒含量分析结果显示,心脏、肝脏、鳃、鳍的AEAdoV相对含量较高,而黏液、皮肤和脾脏的病毒含量相对较低。研究表明,建立的AEAdoV的灵敏度高、特异性强的普通PCR和qPCR检测方法,证实AEAdoV与美洲鳗鲡“出血性烂鳃”病密切相关,且在感染鳗鲡主要组织中都存在。实验结果对于研究AEAdoV的致病性,开展其流行情况和病原学情况分析具有重要意义。

香辛料SYBR GREENⅠ染料实时聚合酶链式反应检测方法的建立及应用

目的:本研究致力于建立香辛料真实性成分的分子鉴定方法,为香辛料的掺假提供有效的鉴定和检测方法,在不同的使用条件下分别实现定性或半定量分析,为建立相关标准提供一定的基础技术支撑。方法:研究采用SYBR GREENⅠ染料实时聚合酶链式反应(polymerase chain reaction,PCR)方法,通过设计特异性引物成功建立了特异性检测八角、茴香、胡椒和花椒的分析技术。结果:该方法能够特异性检测出八角、茴香、胡椒、花椒,Ct值在标准品体积分数0.001%~10%范围内线性关系良好。针对八角标准品,检测限为1%;针对茴香、花椒、胡椒标准品,最检测限均为0.001%。利用该方法对5份实际样本进行检测,分别在样本1中未检测出八角、茴香、胡椒、花椒;样本2、3、5中检测出八角、茴香、胡椒;样本4中检测出八角、茴香、胡椒、花椒。结论:本实验建立的SYBR GREENⅠ染料实时PCR方法灵敏度高、重复性好,可应用于实际样品的检测。

SYBR GreenⅠ染料-实时荧光定量聚合酶链式反应法检测发酵乳中的霉菌和酵母含量

目的建立一种应用SYBR GreenⅠ染料的实时荧光定量聚合酶链式反应法快速检测发酵乳中的霉菌和酵母菌。方法本研究针对霉菌与酵母菌的保守序列设计引物,确定最优反应体系与反应条件。通过熔解曲线以及非目标菌株的检测验证该方法的特异性;通过菌悬液的梯度稀释检测确定该方法的灵敏度;通过菌悬液与发酵乳样品混合后进行检测,确定该方法的检出限,并得到标准曲线。结果该方法能够特异性的检测发酵乳中的霉菌、酵母菌,无交叉反应。该方法检测霉菌、酵母菌的灵敏度均为10^(2) CFU/mL,并且当菌悬液与发酵乳样品混合后,并没有降低该方法的检出限。在10^(2)~10^(6) CFU/mL浓度范围内,菌液浓度的对数值与循环阈值(cyclethreshold,Ct)具有良好的线性关系,可以在市售样品的检测中通过Ct值计算样品中目标菌的含量,更快的判断发酵乳是否被霉菌、酵母菌污染。结论该方法可用于发酵乳中霉菌和酵母菌的快速检测,为发酵乳的食品安全提供了技术保障。

和美乡村视角下的苏南农田“GREEN”水环境治理模式应用研究

随着中国宜居宜业和美乡村建设的不断深入,乡村水环境的治理与保护作为乡村生态文明建设的重要组成部分日益受到社会的重视和关注。在以往大规模灰色基础设施建设的背景下,乡村忽略了对于水生态绿色基础设施的构建,生态格局趋于破碎。农田空间作为农业面源污染的重要源头,结合多学科视角探索农田水环境污染治理新模式具有高度的必要性。立足于苏南地区独特的农田水网体系,紧密结合农业空间生产属性,研究现有农田水环境治理理论方法与实际建设,提炼以“源头绿色—过程净化—末端吸收—生态保育—循环回用”的苏南农田“GREEN”水环境治理模式,探索风景园林学科水治理方法在农田沟渠、河道及湿地等乡村空间的运用路径,并以常州市中天黄金大农场总体规划为例进行实践应用。推动风景园林学科于乡村生产空间的应用模式探索,为未来农业面源污染防治提供参考借鉴。

Does Green Food Certification promote agri-food export quality?Evidence from China

The construction of a food certification system plays a vital role in upgrading export quality, which previous studies have largely overlooked. We match China's industry-level data of Green Food Certification with its HS6-digit export data of agri-food products to quantify the impact of Green Food Certification on export quality. We identify the significant and positive effect of Green Food Certification on export quality. The 2SLS estimation based on instrumental variables and a range of robustness checks confirm the validity and robustness of the benchmark conclusions. Further analysis discloses that Green Food Certification improves export quality by raising agricultural production efficiency and brand premiums. Heterogeneity analysis shows that the effect of Green Food Certification varies across regions, notably improving the quality of agri-food products exported to developed regions and regions with high levels of import supervision. Furthermore, among various product types, Green Food Certification significantly improves the export quality of primary products and products vulnerable to non-tariff measures. The above findings could guide the future development of agri-food quality certification systems, potentially leading to a transformation and promotion of the agri-food trade.

Effect of a Commercial Extract of Green Tea and a Pure Catechin on Two Veillonella strains

The catechin Epigallocatechin-3-O-Gallate (EGCG) which is found in of Green Tea extracts (GTE), displays a variety of microbicidal properties. It is largely believed that EGCG inhibits the growth of cariogenic and periodontopathic bacteria. Objective: In this paper we compared the inhibitory activity of EGCG and a commercial GTE on the growth of Veillonella parvula. Chlorhexidine was used as positive control. Methodology: V. parvula ATCC 10790 and a clinical isolate obtained from a periodontal disease patient were cultured in the presence of EGCG or a commercial GTE, and the measurements of bacterial growth inhibition were compared to the values obtained with 0.12 and 0.2% chlorhexidine. Results: Chlorhexidine inhibited bacterial growth, however in contrast to a previous report, neither EGCG nor the GTE showed any effect on bacterial growth. Conclusions: The data show and confirm that chlorhexidine is a growth inhibitor of V. parvula while EGCG and GTE do not display such effect.

Green Synthesis of Iron(Ⅱ,Ⅲ)-polyphenol Nanoparticles and Their Adsorption of Malachite Green

Three kinds of iron nanoparticles(FeNPs)were prepared via green route based on pomegranate(PG),green tea(GT),and mulberry(ML)extracts under ambient conditions.The obtained materials were characterized by scanning electron microscopy(SEM),transmission electronic microscopy(TEM),X-ray energy-dispersive spectrometer(EDS),X-ray diffraction(XRD),fourier transform infrared spectroscopy(FTIR),and X-ray photoelectron spectroscopy(XPS)techniques.The experimental results show that FeNPs were in the form of amorphous iron(Ⅱ,Ⅲ)-polyphenol complex with different dispersity and morphologies.GT-Fe has the smallest size range of 25-35 nm,PG-Fe has a moderate size-distribution of 30-40 nm,while ML-Fe formed a tuberous net-type with a sheeting structure.PG-Fe displays the highest removal efficiency of 90.2%in 20 min towards cationic dye of malachite green(16.6%by ML-Fe and 69.3%by GT-Fe),which is attributed to its highest polyphenol content,lowest zeta potential,as well as the most Fe^(2+)on the surface of FeNPs.The removal mechanism was mainly induced by electrostatic adsorption based on pH and zeta potential tests.

Establishment of Regional Green Innovation Value Chains:A Case Study on Tackling the Impeded Diffusion of Green Products

The green innovation value chain is a key step in transforming green,innovative scientific,and technological achievements into productive forces.The establishment of green innovation value chains based on value distribution rather than technical conditions can effectively overcome the common bottleneck faced by different nations during their green innovation endeavors,namely,the substitution of conventional products with green alternatives.This study investigates the impeded diffusion of green products and their underlying causes,analyzes the internal structure and mechanism of the green innovation value chain,and explores the establishment of regional green innovation value chains and the models available for value chain upgrading.

Near-infrared cholangiography with intragallbladder indocyanine green injection in minimally invasive cholecystectomy

Laparoscopic cholecystectomy(LC)remains one of the most commonly performed procedures in adult and paediatric populations.Despite the advances made in intraoperative biliary anatomy recognition,iatrogenic bile duct injuries during LC represent a fatal complication and consist an economic burden for healthcare systems.A series of methods have been proposed to prevent bile duct injury,among them the use of indocyanine green(ICG)fluorescence.The most commonly reported method of ICG injection is the intravenous administration,while literature is lacking studies investigating the direct intragallbladder ICG injection.This narrative mini-review aims to assess the potential applications,usefulness,and limitations of intragallbladder ICG fluorescence in LC.Authors screened the available international literature to identify the reports of intragallbladder ICG fluorescence imaging in minimally invasive cholecystectomy,as well as special issues regarding its use.Literature search retrieved four prospective cohort studies,three case-control studies,and one case report.In the three case-control studies selected,intragallbladder near-infrared cholangiography(NIRC)was compared with standard LC under white light,with intravenous administration of ICG for NIRC and with standard intraoperative cholangiography(IOC).In total,133 patients reported in the literature have been administered intragallbladder ICG administration for biliary mapping during LC.Literature includes several reports of intragallbladder ICG administration,but a standardized technique has not been established yet.Published data suggest that NIRC with intragallbladder ICG injection is a promising method to achieve biliary mapping,overwhelming limitations of IOC including intervention and radiation exposure,as well as the high hepatic parenchyma signal and time interval needed in intravenous ICG fluorescence.Evidence-based guidelines on the role of intragallbladder ICG fluorescence in LC require the assessment of further studies and multicenter data collection into large registries.

Rapid Green Synthesis of Silver Nanoparticles by Reishi and Their Antibacterial Activity and Mechanisms

Nanotechnology is a rapidly growing field in biomedical engineering with references to efficiency, safety, and cost-effective approaches. Herein, the objective of this study was to examine an innovative approach to rapidly synthesis silver nanoparticles from an aqueous extract of medicinal mushroom Ganoderma lucidum (also known as reishi). The structural and dimensional dispersion of the biosynthesized silver nanoparticles from reishi was confirmed by UV-Vis spectrophotometer (UV-Vis) and Scanning Electron Microscopy (SEM) analysis. Additionally, the biosynthesized silver nanoparticles from resihi were used to explore their potential antimicrobial activity against Staphylococcus aureus and Micrococcus luteus and Escherichia coli and Klebsiella pneumoniae. The results from this study revealed that the silver nanoparticles mediated by reishi adopted a spherical shape morphology with sizes, less than 100 nm and revealed strong absorption plasmon band at 440 nm. Furthermore, the biosynthesized silver nanoparticles from reishi exhibited antibacterial activity against the tested S. aureus and M. luteus and E. coli and K. pneumoniae by altering their morphology which signifies their biomedical potential.

Structural and Luminescent Properties of Mg_(0.25-x)Al_(2.57)O_(3.79)N_(0.21):xMn^(2+)Green-Emitting Transparent Ceramic Phosphor

A series of spinel-type Mg_(0.25-x)Al_(2.57)O_(3.79)N_(0.21):xMn^(2+)(MgAlON:xMn^(2+))phosphors were synthesized by the solid-state reaction route.The transparent ceramic phosphors were fabricated by pressureless sintering followed by hot-isostatic pressing(HIP).The crystal structure,luminescence and mechanical properties of the samples were systematically investigated.The transparent ceramic phosphors with tetrahedrally coordinated Mn^(2+)show strong green emission centered around 515 nm under blue light excitation.As the Mn^(2+)concentration increases,the crystal lattice expands slightly,resulting in a variation of crystal field and a slight red-shift of green emission peak.Six weak absorption peaks in the transmittance spectra originate from the spin-forbidden ^(4)T_(1)(^(4)G)→^(6)A_(1) transition of Mn^(2+).The decay time was found to decrease from 5.66 to 5.16 ms with the Mn^(2+)concentration.The present study contributes to the systematic understanding of crystal structure and properties of MgAlON:xMn^(2+)green-emitting transparent ceramic phosphor which has a potential application in high-power light-emitting diodes.

Influence mechanism of government subsidy on the green transformation of coal company in China

The optimization of government subsidies to enhance the efficiency of coal companies’green transformation constitutes a critical component in the pursuit of global sustainability.We investigate the influence mechanism of government subsidies on the green transformation using data from the listed coal companies in China from 2007 to 2022.According to our findings and hypothesis testing,previous government subsidies did not have a significant direct impact on coal companies’green transformation.Nevertheless,government subsidies can help coal companies transition to greener practices by promoting innovative green initiatives.Furthermore,we confirmed an indirect route:that government subsidies enable the adoption of low-carbon initiatives,which in turn could facilitate the transition of coal companies towards green practices.In addition,we discovered that the coal company’s digitization will improve this indirect route.Thus,we propose increasing the effectiveness of government subsidies in facilitating coal companies’transition to green practices by focusing on technological advancements and enhancing company digitalization.

A green cross-linking method for the preparation of renewable threedimensional graphene sponges for efficient adsorption of Congo red dye

Graphene-based materials possess significant potential for the treatment of dye wastewater due to their exceptional adsorption properties toward stubborn pollutants.However,their utilization is hindered by high preparation costs,low yields,environmental pollution during synthesis,and challenges in regenerating the adsorbent.This study proposes a novel approach to address these limitations by developing nitrogen-doped three-dimensional(3D)polyvinyl alcohol(PVA)crosslinked graphene sponges(N-PGA)using a cross-linking method with ammonium carbonate.This method offers a relatively mild,environmentally friendly approach.Ammonium carbonate serves as both a reducing and modifying agent,facilitating the formation of the intrinsic structure of N-PGA and acting as a nitrogen source.Meanwhile,PVA is utilized as the cross-linking agent.The results demonstrate that N-PGA exhibits a favorable internal 3D hierarchical porous structure and possesses robust mechanical properties.The measured specific surface area(BET)of N-PGA was as high as406.538 m^(2)·g^(-1),which was favorable for its efficient adsorption of Congo red(CR)dye molecules.At an initial concentration of 50 mg·L^(-1),N-PGA achieved an impressive removal rate of 89.6%and an adsorption capacity of 112 mg·g^(-1)for CR dye.Furthermore,it retained 79%of its initial adsorption capacity after 10 cycles,demonstrating excellent regeneration performance.In summary,the synthesized N-PGA displays remarkable efficacy in the adsorption of CR dye in wastewater,opening up new possibilities for utilizing 3D porous graphene nanomaterials as efficient adsorbents in wastewater treatment.

Green Synthesis of Silver Nanoparticles Using Aqueous Orange and Lemon Peel Extract and Evaluation of Their Antimicrobial Properties

Green synthesis of silver nanoparticles (AgNPs) using aqueous extracts of orange and lemon peels, as a reducing agent, and silver nitrate salts as a source of silver ions is a promising field of research due to the versatility of biomedical applications of metal nanoparticles. In this paper, AgNPs were synthetized at different reaction parameters such as the type and concentration of the extracts, metal salt concentration, temperature, speed stirring, and pH. The antibacterial properties of the obtained silver nanoparticles against E. coli, as well as the physical and chemical characteristics of the synthesized silver nanoparticles, were investigated. UV-Vis spectroscopy was used to confirm the formation of AgNPs. In addition to green biogenic synthesis, chemical synthesis of silver nanoparticles was also carried out. The optimal temperature for extraction was 65˚C, while for the synthesis of AgNPs was 35˚C. The synthesis is carried out in an acidic environment (pH = 4.7 orange and pH = 3.8 lemon), neutral (pH = 7) and alkaline (pH = 10), then for different concentrations of silver nitrate solution (0.5 mM - 1 mM), optimal time duration of the reaction was 60 min and optimal stirring speed rotation was 250 rpm on the magnetic stirrer. The physical properties of the synthesized silver nanoparticles (conductivity, density and refractive index) were also studied, and the passage of laser light through the obtained solution and distilled water was compared. Positive inhibitory effect on the growth of new Escherichia coli colonies have shown AgNPs synthesized at a basic pH value and at a 0.1 mM AgNO3 using orange or lemon peel extract, while for a 0.5 mM AgNO3 using lemon peel extract.

Eutrophication of Jiangsu Coastal Water and Its Role in the Formation of Green Tide

Since 2007,the large-scale green tide caused by Ulva prolifera(U.prolifera)have occurred as a recurrent phenomenon in the southern Yellow Sea of China.Field surveys and satellite remote sensing showed that the small scattered patches of green tide algae were first observed along the Porphyra agriculture area of the Subei Shoal in late April.In this study,we attempted to identify the role of eutrophication in the origin of the green tide in the Subei Shoal and its adjacent area.Subei Shoal and its adjacent area are characterized by rich nutrients,especially NO_(3)^(-)-N,NH_(4)^(+)-N,PO_(4)^(3-)-P,and other bioavailable components(such as urea-N and amino acids).In the spring of 2017,the average concentrations of NO_(3)^(-)-N were 19.01±11.01μmolL^(-1),accounting for 86.68%of the dis-solved inorganic nitrogen(DIN).In addition,the average concentration of NH4^(+)-N was 2.51±1.60μmolL^(-1).PO_(4)^(3-)-P had an average concentration of 0.14±0.13μmolL-1.The average concentrations of urea-N and total hydrolyzed amino acids(THAA)were 1.73±1.36μmolL^(-1)and 1.33±0.80μmolL^(-1),respectively.Rich nutritive substances play a key role in the rapid production of U.prolifera and make the Jiangsu coastal water an incubator for green tide.

Cost Control Strategies for Hubei General Highway Greening Project

In order to respond to the national policy of focusing on the service of building a new development pattern and promoting high-quality development,and to reduce the cost of greening projects while ensuring the effect of highway landscape,a process and cost control points of highway greening design,construction,and maintenance are summarized through a review of the literature.Additionally,this paper examines the attributes of highway greening and proposes cost control strategies that are aligned with these attributes.It is proposed that the implementation of cost control strategies for highway greening should commence at the project establishment phase,with the objective of establishing a comprehensive and effective cost management control system.While guaranteeing the greening landscape effect and the duration of the project,it is essential to regulate the crucial nodes in each phase of the design,construction,and maintenance process.Furthermore,it is vital to facilitate close collaboration between all parties involved,thereby reducing costs,conserving resources,and lowering energy consumption.This approach can also lead to enhanced economic and social benefits for highway greening projects.