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动物源弯曲杆菌免疫显性抗原鉴定及间接ELISA检测方法的建立
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作者 杨欢 王策 +6 位作者 王玉飞 赵晶 张会强 曹易博 姬钰浩 邓兴梅 张辉 《中国畜牧兽医》 北大核心 2026年第3期1418-1433,共16页
【目的】筛选胎儿弯曲杆菌(Campylobacter fetus)、空肠弯曲杆菌(Campylobacter jejuni)血清免疫显性抗原,初步建立动物弯曲杆菌病血清学ELISA诊断方法。【方法】利用生物信息学分析软件对候选抗原进行分析筛选,用pET-28a(+)原核表达载... 【目的】筛选胎儿弯曲杆菌(Campylobacter fetus)、空肠弯曲杆菌(Campylobacter jejuni)血清免疫显性抗原,初步建立动物弯曲杆菌病血清学ELISA诊断方法。【方法】利用生物信息学分析软件对候选抗原进行分析筛选,用pET-28a(+)原核表达载体构建重组质粒,诱导表达并纯化重组蛋白。通过SDS-PAGE和His标签抗体Western blotting检测蛋白表达情况。制备兔源弯曲杆菌多克隆抗体并将其作为Western blotting一抗验证抗原与多克隆抗体反应原性。建立间接ELISA检测方法,优化抗原包被浓度、封闭液及封闭时间、一抗和二抗稀释度及反应时间等反应条件,确定临界值,评价其特异性、灵敏性、重复性。【结果】利用生物信息学分析软件筛选出胎儿弯曲杆菌抗原SapA、空肠弯曲杆菌抗原FlaA。原核表达纯化后,Western blotting检测SapA蛋白大小为101 ku,FlaA蛋白大小为59 ku。制备的胎儿弯曲杆菌兔多克隆抗体血清效价为1∶64000,空肠弯曲杆菌兔多克隆抗体效价为1∶128000。Western blotting结果表明,抗原SapA、FlaA特异性识别弯曲杆菌多克隆抗体,具有良好的反应原性。以纯化的SapA蛋白作为包被抗原,确定的最优包被浓度为2.5μg/mL,封闭液为5%BSA,封闭时间30 min,一抗、二抗稀释度分别为1∶500和1∶8000,孵育时间分别为30和90 min。以纯化的FlaA蛋白作为包被抗原,确定最优包被浓度为0.4μg/mL,封闭时间1 h,其余条件与前者一致。二者与结核分枝杆菌(MTB)、副结核分枝杆菌(MAP)及布鲁氏菌(Brucella)均无交叉反应,特异性强。SapA间接ELISA方法的灵敏性最低达到1∶6400,FlaA间接ELISA方法的灵敏性达到1∶12800。批内和批间重复性试验变异系数均<10%,证明其重复性好。【结论】本研究成功鉴定了胎儿弯曲杆菌免疫显性抗原SapA、空肠弯曲杆菌免疫显性抗原FlaA,建立了动物弯曲杆菌病血清学间接ELISA诊断方法,为弯曲杆菌病防控提供了有效的诊断抗原及快速诊断方法。 展开更多
关键词 弯曲杆菌 免疫显性抗原 原核表达 Western blotting 间接ELISA
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The Cultivation of Caring Thinking in Education for the Re-Creation of the Democratic Human Being
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作者 Alexandros Theodoridis 《Philosophy Study》 2026年第1期1-17,共17页
In a world growing ever more cruel and violent as nihilism becomes generalized,a world in which the disintegration of the human assumes dimensions that threaten its very cohesion,the demand for the humanization of the... In a world growing ever more cruel and violent as nihilism becomes generalized,a world in which the disintegration of the human assumes dimensions that threaten its very cohesion,the demand for the humanization of the Western individual asserts itself as an urgent historical task.Among the institutions implicated in this process,education assumes a central role,as it is within it that wonder and critical thinking are gradually eliminated,together with the demand for shared deliberation,coherent meaning,and the tragic dimension of human existence,while an education of cruelty comes to permeate it from end to end.In order to prevent the full entrenchment of evil in the social world,from which only destruction can be expected,a decisive role could be played by caring thinking that would be coordinated with corresponding action and would come to deeply characterize the educational institution.Such a form of thinking was proposed in the second half of the twentieth century and took flesh and bones within the framework of the Philosophy for Children movement.Its contemporary application within educational practice could reactivate phronesis(practical wisdom)and contribute to enabling the Western human being to re-create themselves as a democratic human being-one who desires the creation of a world of freedom,equality,and justice,and who acts accordingly to bring it about:the creation of a compassionate and prudent world,the preservation of which would be the responsibility of all. 展开更多
关键词 Western world EDUCATION CRUELTY VIOLENCE caring thinking democratic human being
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Analysis and Modelling of Salt-related Deformation in the Western Sichuan Foreland Basin,China
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作者 QIU Dengfeng HU Zongquan +2 位作者 John J.WALSH GUO Jinrui ZHANG Zhongming 《Acta Geologica Sinica(English Edition)》 2026年第1期265-285,共21页
The Western Sichuan Foreland Basin(WSFB)in South China,a prolific hydrocarbon province,exhibits complex structural deformation influenced by Triassic salt tectonics.This paper integrates seismic data and well data to ... The Western Sichuan Foreland Basin(WSFB)in South China,a prolific hydrocarbon province,exhibits complex structural deformation influenced by Triassic salt tectonics.This paper integrates seismic data and well data to elucidate the role of Middle-Lower Triassic evaporite layers in shaping basin structures,focusing on Xinchang Tectonic Zone(XTZ).Salt layers facilitated decoupled deformation between supra-and sub-salt sequences,forming salt pillows and fault-related folds.Three distinct structural trends were identified in XTZ.Key findings reveal that salt thickness variations correlate with deformation styles:thicker salt promoted detachment folding,while thinner salt led to hard-linked fault systems.Sub-salt E-NE trending reverse faults formed horsetail terminations associated with the Pengzhou faults(PzF),deviating from the primary Longmenshan thrust belt(LmsTB)orientation.Structural evolution occurred in three stages:(1)Indosinian salt deposition and foreland basin initiation;(2)Yanshanian eastward propagation of thrust systems with salt-driven detachment folding;(3)Himalayan reactivation overprinting earlier structures with sub-NS trending folds.This work establishes a direct link between salt layers and structural traps,demonstrating how salt acted as a critical detachment layer during multi-stage compression.Results provide insights into the gas exploration of the Late Triassic Xujiahe Formation,emphasizing the importance of salt-influenced deformation in foreland basin systems. 展开更多
关键词 salt structure FAULT horsetail terminations foreland basin western Sichuan
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Bridging Dreams,Building Futures
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《ChinAfrica》 2026年第3期14-15,共2页
China-Zimbabwe collaboration is creating new opportunities for innovation and development of youth Zimbabwe’s development has long faced obstacles linked to Western economic sanctions,measures that many view as unjus... China-Zimbabwe collaboration is creating new opportunities for innovation and development of youth Zimbabwe’s development has long faced obstacles linked to Western economic sanctions,measures that many view as unjust or unlawful.Yet,despite these constraints,Zimbabwe remains a nation rich in natural resources and endowed with a highly educated population,with its youth standing out as a particularly dynamic force. 展开更多
关键词 innovation DEVELOPMENT natural resources economic sanctions development youth obstacles western economic sanctionsmeasures DREAMS
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Climate and human drivers of surface water changes in the Zavkhan River-Khyargas Lake Basin,western Mongolia
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作者 BATSUREN Dorjsuren VALERY A.Zemtsov +4 位作者 ERDENEBAYAR Bavuu SANDELGER Dorligjav YAN Denghua GAO Hongkai ALTANBOLD Enkhbold 《Journal of Geographical Sciences》 2026年第1期255-280,共26页
This study investigates climate-and human-induced hydrological changes in the Zavkhan River-Khyargas Lake Basin,a highly sensitive arid and semi-arid region of Central Asia.Using Mann-Kendall,innovative trend analysis... This study investigates climate-and human-induced hydrological changes in the Zavkhan River-Khyargas Lake Basin,a highly sensitive arid and semi-arid region of Central Asia.Using Mann-Kendall,innovative trend analysis,and Sen's slope estimation methods,historical climate trends(1980-2100)were analyzed,while land cover changes represented human impacts.Future projections were simulated using the MIROC model with Shared Socioeconomic Pathways(SSPs)and the Tank model.Results show that during the past 40 years,air temperature significantly increased(Z=3.93^(***)),while precipitation(Z=-1.54^(*))and river flow(Z=-1.73^(*))both declined.The Khyargas Lake water level dropped markedly(Z=-5.57***).Land cover analysis reveals expanded cropland and impervious areas due to human activity.Under the SSP1.26 scenario,which assumes minimal climate change,air temperature is projected to rise by 2.0℃,precipitation by 21.8 mm,and river discharge by 1.61 m^(3)/s between 2000 and 2100.These findings indicate that both global warming and intensified land use have substantially altered hydrological and climatic processes in the basin,highlighting the vulnerability of western Mongolia's water resources to combined climatic and anthropogenic influence. 展开更多
关键词 arid zone semi-arid zone climate change land cover trend analysis western Mongolia
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Aims&Scope
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《Contemporary Social Sciences》 2026年第1期F0002-F0002,共1页
Contemporary Social Sciences publishes outstanding research in the field of social sciences in China and also includes high-quality research work by foreign scholars on the development of China’s western regions and ... Contemporary Social Sciences publishes outstanding research in the field of social sciences in China and also includes high-quality research work by foreign scholars on the development of China’s western regions and its reform and opening up.The aim is to help promote China’s academic achievements to the world and give China a stronger voice in the global community of social sciences.The scopes of research fields for consideration include,but not limited to political science,economics,literature,linguistics,journalism and communication,education,sociology,philosophy,history,law,and interdisciplinary research. 展开更多
关键词 global community academic achievements ECONOMICS social sciences China political science reform opening up western regions
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Retraction:Long Noncoding RNA PVT1 Promotes Melanoma Progression via Endogenous Sponging miR-26b
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第3期766-766,共1页
The published article titled“Long Noncoding RNA PVT1 PromotesMelanoma Progression via Endogenous Sponging miR-26b”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.675–681.DOI:10.3727/096504017X14920318... The published article titled“Long Noncoding RNA PVT1 PromotesMelanoma Progression via Endogenous Sponging miR-26b”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.675–681.DOI:10.3727/096504017X14920318811730 URL:https://www.techscience.com/or/v26n5/56680 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 cellular datawhere protein bands cellular data PVT long noncoding RNA MELANOMA miR b western blot
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Unravelling tree diversity patterns and responses to environmental gradients in a tropical forest landscape of the Western Ghats
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作者 Naveen Babu Kanda Ashaq Ahmad Dar +2 位作者 Kurian Ayushi Ayyappan Narayanan Narayanaswamy Parthasarathy 《Plant Diversity》 2026年第1期92-106,共15页
Understanding spatial patterns of plant species diversity and the factors(e.g.,climate and human)that drive these patterns is essential for biodiversity conservation.We used data from 1700.1-ha forest plots in the She... Understanding spatial patterns of plant species diversity and the factors(e.g.,climate and human)that drive these patterns is essential for biodiversity conservation.We used data from 1700.1-ha forest plots in the Shettihalli tropical forest landscape of the Western Ghats biodiversity hotspot,India,to analyse tree community composition and the drivers ofα-diversity(Shannon)andβ-diversity(LCBD).Compositional patterns were visualized using Non-Metric Multidimensional Scaling(NMDS),and hybrid feature selection with structural equation modeling(SEM)was employed to evaluate the direct and indirect effects of environmental variables on diversity.NMDS identified four distinct forest types in the Shettihalli landscape:semi-evergreen,dry deciduous,moist deciduous,and plantation forests,each with distinct plant composition.Shannon diversity and ecological uniqueness was significantly higher in semi-evergreen forest than in deciduous forest plots.The SEMs explained about 79%and 39–45%of the variation inα-diversity andβ-diversity.Our analysis indicated that current diversity patterns result from multiple processes,with structure,disturbance,and edaphic parameters exerting the strongest direct and indirect effects onα-diversity.β-diversity,in contrast,was largely influenced by climate,topography,stand structure,and edaphic factors.Overall,our findings indicate that various factors(e.g.,climate,topography,and human disturbance)interact to shape tree diversity patterns in tropical forests.These findings will help develop unique conservation and management strategies for distinct forest types in tropical forest ecosystems. 展开更多
关键词 Alpha diversity Beta diversity Machine learning Structural equation modeling Vegetation patterns Western Ghats
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Mechanisms of ENSO's cross-seasonal modulation of winter–spring atmospheric river activity over East Asia
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作者 Yuliang Zhou Wentao Jia +1 位作者 Weimin Zhang Huizan Wang 《Atmospheric and Oceanic Science Letters》 2026年第1期66-71,共6页
In this study,based on MERRA-2 reanalysis data and a multi-algorithm integrated atmospheric river(AR)iden-tification method,the authors reveal the cross-seasonal regulation mechanism of El Niño-Southern Oscillati... In this study,based on MERRA-2 reanalysis data and a multi-algorithm integrated atmospheric river(AR)iden-tification method,the authors reveal the cross-seasonal regulation mechanism of El Niño-Southern Oscillation(ENSO)on winter-spring AR activities in East Asia.The results show that ENSO asymmetrically modulates AR ac-tivity through teleconnection and hysteresis effects,and has significant enhancement/inhibition effects on ARs in different regions.At the onset of El Niño,enhanced southwesterly flow at the western edge of the western Pacific subtropical high(WPSH)leads to enhanced AR activity in the western Pacific,and anomalous southerly winds in the Indian Ocean promote northward transport of water vapor in the Arabian Sea and Bay of Bengal.With a three-month lag,the weakening and eastward retreat of the WPSH weakens the low-latitude AR activity,but persistent southerly winds in the Bay of Bengal maintain the AR activity over Southwest China.The mid-to high-latitude AR response exhibits delayed dynamics,initially dominated by the synergistic effect of the southward deviation of the upper-air rapids and the low-level convergence(double-rapid-flow effect)and later modulated by the Pacific-North American teleconnection(PNA)-triggered East Asian ridge,which enhances the precipitation efficiency through prolonged frontal activity and enhanced cold-warm airmass convergence.Overall,El Niño promotes the development of low-and midlatitude AR activity in East Asia,while La Niña promotes(maritime continental)AR activity in the tropics.This study establishes the“ENSO teleconnection→circulation adjust-ment→East Asian AR response”chain,revealing a cross-seasonal lagged response mechanisms of East Asian AR activity,and provides a theoretical basis for winter and spring climate prediction and extreme precipitation forecasting. 展开更多
关键词 ENSO Atmospheric river Western Pacific subtropical high East Asia Lagged response
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Retraction:Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis and Metastasis of Non-Small Cell Lung Cancer by Upregulating miR-613
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第3期767-767,共1页
The published article titled“Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis andMetastasis of Non-Small Cell Lung Cancer by Upregulating miR-613”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.... The published article titled“Long Noncoding RNA(lncRNA)HOTAIR Affects Tumorigenesis andMetastasis of Non-Small Cell Lung Cancer by Upregulating miR-613”has been retracted from Oncology Research,Vol.26,No.5,2018,pp.725–734.DOI:10.3727/096504017X15119467381615 URL:https://www.techscience.com/or/v26n5/56685 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 non small cell lung cancer cellular datawhere Hotair METASTASIS long noncoding RNA TUMORIGENESIS western blot MIR
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Journey to the East
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作者 《China Today》 2026年第3期16-19,共4页
Chinese President Xi Jinping receives a steady stream of Western leaders over the past two months,demonstrating China’s role as a reliable partner on the international stage.SINCE the start of 2026,Chinese President ... Chinese President Xi Jinping receives a steady stream of Western leaders over the past two months,demonstrating China’s role as a reliable partner on the international stage.SINCE the start of 2026,Chinese President Xi Jinping has welcomed a succession of leaders from Western countries.From British Prime Minister Keir Starmer and Canadian Prime Minister Mark Carney to the leaders of Ireland and Finland. 展开更多
关键词 Chinese President Xi Jinping western leaders reliable partner international stage
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Ningxia Museum
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《China Today》 2026年第2期76-79,共4页
At the Ningxia Museum,visitors can dive into the history of the mysterious Western Xia Dynasty,as well as cultural and ethnic exchanges that occured along the ancient Silk Road centuries ago.THE area of present-day Ni... At the Ningxia Museum,visitors can dive into the history of the mysterious Western Xia Dynasty,as well as cultural and ethnic exchanges that occured along the ancient Silk Road centuries ago.THE area of present-day Ningxia Hui Autonomous Region in northwest China was a vital corridor of the ancient Silk Road.Around a thousand years ago,Yuan Hao,leader of the Tangut people,established the Western Xia(also known as Xixia)Dynasty(1038-1227)on this land,which coexisted with the Song,Liao,and Jin dynasties across China for nearly 200 years. 展开更多
关键词 cultural ethnic exchanges Silk Road Ningxia Museum Yuan Hao Western Xia Dynasty Cultural Exchange
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Retraction: MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase
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作者 Oncology Research Editorial Office 《Oncology Research》 2026年第1期622-622,共1页
The published article titled“MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase”has been retracted from Oncology Research,Vol.25,No.8,2017,pp.1231–1243.DOI:10.3... The published article titled“MicroRNA-148a Acts as a Tumor Suppressor in Osteosarcoma via Targeting Rho-Associated Coiled-Coil Kinase”has been retracted from Oncology Research,Vol.25,No.8,2017,pp.1231–1243.DOI:10.3727/096504017X14850134190255 URL:https://www.techscience.com/or/v25n8/56908 Following the publication,concerns have been raised about a number of figures in this article.An unexpected area of similarity was identified in terms of the cellular data,where the results from differently performed experiments were intended to have been shown,although the areas immediately surrounding this area featured comparatively different distributions of cells.In addition,the western blots in this article were presented with atypical,unusually shaped and possibly anomalous protein bands in many cases. 展开更多
关键词 cellular datawhere tumor suppressor western blots rho associated coiled coil kinase microrna cellular data OSTEOSARCOMA figures
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Corrigendum
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《Neural Regeneration Research》 2026年第3期922-922,共1页
Corrigendum:Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway https://doi.org/10.4103/NRR.NRR-D-25-00562 In the article titled“Epalrestat protect... Corrigendum:Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway https://doi.org/10.4103/NRR.NRR-D-25-00562 In the article titled“Epalrestat protects against diabetic peripheral neuropathy by alleviating oxidative stress and inhibiting polyol pathway,”published on pages 345-351 in Issue 2,Volume 11 of Neural Regeneration Research(Li et al.,2016),the Western blot bands in Figure 2A are incorrect. 展开更多
关键词 western blot bands diabetic peripheral neuropathy alleviating oxidative stress oxidative stress polyol pathway EPALRESTAT
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Journal of Acupuncture and Tuina Science Instructions for Authors
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作者 CHEN Hanping WU Huangan 《Journal of Acupuncture and Tuina Science》 2026年第1期I0003-I0004,共2页
The Journal of Acupuncture and Tuina Science,a peerreview journal with an international scope(ISSN 1672-3597,CN 31-1908/R,Bimonthly),is indexed in Emerging Sources Citation Index(ESCI),Western Pacific Region Index Med... The Journal of Acupuncture and Tuina Science,a peerreview journal with an international scope(ISSN 1672-3597,CN 31-1908/R,Bimonthly),is indexed in Emerging Sources Citation Index(ESCI),Western Pacific Region Index Medicus(WPRIM),Springer-Verlag GmbH Database,Index of Copernicus(IC),Chemical Abstracts(CA),Chinese Science Citation Database(CSCD),and Chinese Scientific and Technical Papers and Citations Database(CSTPCD).You can search the text on https://link.springer.com/journal/11726. 展开更多
关键词 International Scope ESCI WPRIM TUINA ACUPUNCTURE acupuncture tuina emerging sources citation index esci western Springer Verlag GMBH
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Neoproterozoic Overprinting on the Archean Western Dharwar Craton,Southern India:Records from LA-ICP-MS U-Pb Geochronology of Rutile and Monazite
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作者 HU Yuhua ZHAO Yan +5 位作者 LU Junsheng GOU Longlong PMGEORGE AO Wenhao ZHANG Chengli ZHAI Mingguo 《Acta Geologica Sinica(English Edition)》 2026年第1期82-98,共17页
This study provides new evidence for a significant Neoproterozoic tectono-thermal event in the southern Dharwar Craton,based on U-Pb dating of rutile and monazite from metamorphic rocks of the Sargur Group.Trace eleme... This study provides new evidence for a significant Neoproterozoic tectono-thermal event in the southern Dharwar Craton,based on U-Pb dating of rutile and monazite from metamorphic rocks of the Sargur Group.Trace element analysis of rutile in garnet-bearing amphibolite,amphibolite,garnet-bearing staurolite-kyanite schist,and grunerite-garnet schist suggests that their protoliths were primarily basaltic rocks,sediments with mafic component,pelitic sediments,and mixed sediments with both mafic and felsic components,respectively.Rutile U-Pb ages of 625±22 Ma(garnet-bearing amphibolite),752±5.1 Ma(amphibolite),836±4 Ma to 663±26 Ma(garnet-bearing staurolite kyanite schist)and 860±13 Ma(grunerite-garnet schist),along with a monazite U-Pb age of 789±4.4 Ma(garnet-bearing staurolite kyanite schist),record a significant Neoproterozoic(860-625 Ma)tectono-thermal event along the southern margin of the western Dharwar Craton.This event is most likely associated with aborted rift-related alkaline magmatism along the northern margin of the Southern Granulite Terrane(SGT)and the southern margin of the Dharwar Craton.Given the lack of direct evidence for the assembly of the Rodinia supercontinent in the Dharwar Craton and SGT,this Neoproterozoic event is unlikely to be related to the breakup of Rodinia. 展开更多
关键词 rutile and monazite U-Pb ages NEOPROTEROZOIC Sargur area western Dharwar Craton southern India
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高精度生物检测技术的系统化整合:温控扩展、荧光灵敏度提升与定量标准化研究进展 被引量:1
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作者 杨坤鹏 赵晖 +4 位作者 王雷 王哲 蔡承伦 吕佩昀 王宝 《吉林医学》 2025年第6期1466-1469,共4页
生物检测技术是现代生物医学研究和临床诊断的基础,其精度和可靠性直接影响研究结果的科学性和诊断结果的准确性。随着精准医疗和个体化治疗的快速发展,对生物检测技术的精度要求日益提高。高精度生物检测技术的系统化整合不仅需要优化... 生物检测技术是现代生物医学研究和临床诊断的基础,其精度和可靠性直接影响研究结果的科学性和诊断结果的准确性。随着精准医疗和个体化治疗的快速发展,对生物检测技术的精度要求日益提高。高精度生物检测技术的系统化整合不仅需要优化各项技术的关键参数,还需要建立统一的标准化体系,以确保不同实验室、不同设备之间结果的可比性和一致性。 展开更多
关键词 PCR 流式细胞仪 WESTERN印迹 生物检测技术 系统化整合
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马疱疹病毒1型对BHK-21细胞自噬影响的研究
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作者 玄玉莹 庞涛 +5 位作者 付婧 丁玉林 王文龙 高娃 祁咏华 王金玲 《中国预防兽医学报》 北大核心 2025年第10期1073-1079,共7页
为研究马疱疹病毒1型(EHV-1)感染仓鼠肾纤维细胞(BHK-21)后对细胞自噬功能的影响,本实验首先通过不同感染量(MOI)的EHV-1感染BHK-21细胞48 h后,利用Western blot检测自噬蛋白LC3 II和LC3 I的表达情况,计算LC3 II/LC3 I值;以不同浓度的... 为研究马疱疹病毒1型(EHV-1)感染仓鼠肾纤维细胞(BHK-21)后对细胞自噬功能的影响,本实验首先通过不同感染量(MOI)的EHV-1感染BHK-21细胞48 h后,利用Western blot检测自噬蛋白LC3 II和LC3 I的表达情况,计算LC3 II/LC3 I值;以不同浓度的雷帕霉素(Rapa)和3-甲基腺嘌呤(3-MA)分别作用于BHK-21细胞,采用CCK-8法检测细胞活性,筛选最适药物浓度(Rapa 0.1μmol/L,3-MA 0.5μmol/L)后,经Western blot检测分析EHV-1感染BHK-21细胞与最适药物浓度作用后BHK-21细胞的LC3 II/LC3 I值。结果显示,不同感染剂量EHV-1感染BHK-21细胞48 h后,感染组LC3 II/LC3 I值极显著低于对照组(P<0.001);EHV-1感染组LC3 II/LC3 I值显著低于对照组(P<0.05),且极显著低于Rapa处理组(P<0.001),而与3-MA组无显著差异。利用EHV-1感染BHK-21细胞后,采用Western blot检测不同时间点自噬蛋白LC3和P62的表达情况,并计算LC3 II/LC3 I与P62/GAPDH值;将EHV-1感染过表达LC3B的BHK-21细胞,利用激光共聚焦显微镜观察感染后的聚集情况;以MOI 1 EHV-1感染BHK-21细胞后,利用透射电镜观察不同时间点的自噬体。Western blot结果显示,EHV-1感染BHK-21细胞12 h后LC3 II/LC3 I值均极显著高于对照组(P<0.05),24 h~60 h时LC3 II/LC3 I值均极显著低于对照组(P<0.01);12 h~36 h时P62/GAPDH值显著高于对照组(P<0.05);48 h、60 h的P62/GAPDH值与对照组无显著差异。激光共聚焦显微镜观察可见,感染EHV-112 h后的BHK-21细胞中GFP-LC3B聚集量多于24 h、36 h时的感染组和12 h时的对照组;24 h和36 h感染组的GFP-LC3B聚集少于同一时间的对照组。透射电镜观察结果显示,EHV-1感染BHK-21细胞12 h后,BHK-21细胞中的自噬体明显多于对照组;而感染36 h时BHK-21细胞中自噬体的数量少于对照组和感染12 h自噬体的数量。激光共聚焦和透射电镜观察结果与Western blot检测结果一致。上述结果首次证实,EHV-1感染BHK-21细胞12 h促进细胞自噬,24 h后抑制细胞自噬。本研究为深入探究细胞自噬对EHV-1复制和感染的调控机制奠定基础。 展开更多
关键词 马疱疹病毒1型 BHK-21细胞 细胞自噬 Western blot 透射电镜
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自体黄韧带干预TGF-β1/Smad3信号通路抑制兔硬膜外纤维瘢痕形成的实验研究
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作者 王鹏 张德宝 +5 位作者 张杨 张海斌 时嘉 张少杰 李树文 吴一民 《中国临床解剖学杂志》 北大核心 2025年第1期69-75,共7页
目的通过对不同组别的兔分别行椎板切除术,对比其各组术后3、6周TGF-β1、Smad3蛋白表达量以及两者相应的mRNA表达水平,从而探讨自体黄韧带抑制腰椎术后硬膜外纤维瘢痕形成与TGF-β1/Smad3信号通路的关系。方法将于内蒙古医科大学实验... 目的通过对不同组别的兔分别行椎板切除术,对比其各组术后3、6周TGF-β1、Smad3蛋白表达量以及两者相应的mRNA表达水平,从而探讨自体黄韧带抑制腰椎术后硬膜外纤维瘢痕形成与TGF-β1/Smad3信号通路的关系。方法将于内蒙古医科大学实验动物中心购买的48只6~8月龄的日本大白兔,分为保留黄韧带组,自体脂肪回置组,不保留黄韧带组,对不同组别进行手术造模。术后对每组间的实验动物分别饲养3、6周后进行处死(3、6周各处死8只)。采用Western blot蛋白定量分析、RTPCR技术测定各组样本中的TGF-β1、Smad3蛋白含量及mRNA表达水平,并将以上数据分别进行组间比较,分析其差异性。结果①Western blot检测结果:3、6周保留组TGF-β1、Smad3蛋白表达量小于不保留组和脂肪组(P<0.05),不保留组和脂肪组之间并未发现明显差异。②RT-PCR结果显示:对于TGF-β1、Smad3两者的mRNA表达量而言,3、6周保留组明显小于不保留组和脂肪组(P<0.05),不保留组和脂肪组之间并未见明显差异。结论腰椎手术术后,会形成不同程度的硬膜外纤维瘢痕。若在术中保留自体黄韧带,可减少硬膜外成纤维细胞的形成,其机制与TGF-β1/Smad3信号通路有关。 展开更多
关键词 黄韧带 纤维瘢痕 TGF-β1/Smad3信号通路 Western blot RT-PCR技术
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条件性胰岛β细胞HLF基因敲除小鼠构建与鉴定
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作者 侯梦龙 齐心语 +4 位作者 吴建凤 廖启超 马杰 周磊 李一星 《安徽医科大学学报》 北大核心 2025年第8期1432-1439,共8页
目的为了探究肝白血病因子(HLF)在糖尿病中的作用机制,构建条件性胰岛β细胞HLF基因敲除小鼠动物模型。方法在细胞水平上,通过CCK-8试验验证HLF抑制或过表达对MIN6细胞增殖的影响。通过RT-qPCR和Western blot,分别在mRNA水平和蛋白水平... 目的为了探究肝白血病因子(HLF)在糖尿病中的作用机制,构建条件性胰岛β细胞HLF基因敲除小鼠动物模型。方法在细胞水平上,通过CCK-8试验验证HLF抑制或过表达对MIN6细胞增殖的影响。通过RT-qPCR和Western blot,分别在mRNA水平和蛋白水平检测HLF抑制或过表达效果。将HLF flox/flox转基因小鼠与Pdx1-Cre+/-小鼠(C57BL/6J)杂交繁育,获得子代小鼠。利用PCR方法鉴定小鼠基因型,通过RT-qPCR技术和Western blot检测HLF基因在胰岛β细胞敲除小鼠(HLF flox/flox Cre+/-)和对照小鼠(HLF flox/flox)中的mRNA水平和蛋白水平表达差异,验证敲除效果。同时取2组小鼠的胰岛组织制作石蜡切片并进行苏木精-伊红(HE)染色分析。结果HLF基因抑制或过表达对MIN6细胞增殖无显著影响。在MIN6细胞中抑制HLF基因,其mRNA表达水平较对照组下降了74%,蛋白表达水平较对照组下降了60%;过表达HLF基因后,其mRNA表达水平为对照组的2.13倍,蛋白表达水平为对照组的1.8倍。敲除小鼠的HLF基因mRNA表达水平较对照组下降了89%,蛋白表达水平较对照组下降了65%。HE染色结果显示:敲除小鼠胰岛组织内细胞形态与对照小鼠无明显差异。抑制HLF使MIN6细胞中糖原含量提高约20%。结论成功构建HLF基因敲除小鼠,为研究HLF在糖尿病发病机制中的作用提供了动物模型。 展开更多
关键词 肝白血病因子 条件性胰岛β细胞 RT-QPCR Western blot 基因敲除 C57BL/6J小鼠
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