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Nanobodies against African swine fever virus p72 and CD2v proteins as reagents for developing two cELISAs to detect viral antibodies

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Virologica Sinica 2024年第3期39卷 478-489页

作者： jiahong zhu Qingyuan Liu Liuya Li Runyu Zhang Yueting Chang Jiakai Zhao Siyu Liu Xinyu Zhao Xu Chen Yani Sun Qin Zhao Department of Preventive Veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYangling Observing and Experimental Station of National Data Center of Animal HealthMinistry of AgricultureYangling712100China

African swine fever virus(ASFV)poses a significant threat to the global swine ***,there are no effective vaccines or treatments available to combat ASFV infection in *** primary means of controlling the spread of the disease is through rapid detection and subsequent elimination of infected ***,a lower virulent ASFV isolate with a deleted EP402R gene(CD2v-deleted)has been reported in China,which further complicates the control of ASFV infection in pig ***,an EP402R-deleted ASFV variant has been developed as a potential live attenuated vaccine candidate ***,it is crucial to develop detection methods that can distinguish wild-type and EP402R-deleted ASFV *** this study,two recombinant ASFV-p72 and-CD2v proteins were expressed using a prokaryotic system and used to immunize Bactrian ***,eight nanobodies against ASFV-p72 and ten nanobodies against ASFV-CD2v were *** the production of these nanobodies with horse radish peroxidase(HRP)fusion proteins,the ASFV-p72-Nb2-HRP and ASFV-CD2v-Nb22-HRP fusions were selected for the development of two competitive ELISAs(cELISAs)to detect anti-ASFV *** two cELISAs exhibited high sensitivity,good specificity,repeatability,and *** coincidence rate between the two cELISAs and commercial ELISA kits was 98.6%and 97.6%,***,the two cELISA for detecting antibodies against ASFV demonstrated ease of operation,a low cost,and a simple production *** two cELISAs could determine whether pigs were infected with wild-type or CD2v-deleted ASFV,and could play an important role in monitoring ASFV infections in pig farms.

关键词： African swine fever virus(ASFV) ASFV-p72 ASFV-CD2v Nanobody-HRP Competitive ELISA

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Two nanobody‑based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus

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Animal Diseases 2024年第2期4卷 98-114页

作者： Xu Chen Yueting Chang Lu Zhang Xinyu Zhao Zhihan Li Zhijie Zhang Pinpin Ji Qingyuan Liu Jiakai Zhao jiahong zhu Baoyuan Liu Xinjie Wang Yani Sun Qin Zhao Department of Preventive Veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYanglingShaanxi 712100China Shenzhen Branch Guangdong Laboratory of Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of Agriculture and Rural AffairsAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518100China

Porcine reproductive and respiratory syndrome virus(PRRSV)infection causes significant economic loss to the global pig *** 1 and 2 PRRSV(PRRSV-1 and-2)infections have been reported in China,Europe and *** accurate prevention,nanobodies were first used as diagnostic reagents for PRRSV *** this study three nanobodies targeting both PRRSV-1 and-2,two targeting PRRSV-1 and three targeting PRRSV-2,were screened and *** develop two competitive ELISAs(cELISAs),the g1-2-PRRSV-Nb3-HRP nanobody was chosen for the g1-2-cELISA,to detect common antibodies against PRRSV-1 and-2,and the g1-PRRSV-Nb136-HRP nanobody was chosen for the g1-cELISA,to detect anti-PRRSV-1 *** two cELISAs were developed using PRRSV-1-N protein as coating antigen,and the amounts for both were 100 ng/*** optimized dilution of testing pig sera was 1:20,the optimized reaction times were 30 min,and the colorimetric reaction times were 15 ***,the cut-off values of the g1-2-cELISA and g1-cELISA were 26.6%and 35.6%,*** of them have high sensitivity,strong specificity,good repeatability,and *** addition,for the 1534 clinical pig sera,an agreement rate of 99.02%(Kappa values=0.97)was determined between the g1-2-cELISA and the commercial IDEXX ELISA *** the g1-cELSIA,it can specifically detect anti-PRRSV-1 antibodies in the clinical pig ***,combining two nanobody-based cELISAs can differentially detect antibodies against PRRSV-1 and-2.

关键词： PRRSV Competitive ELISA Nanobody Antigen epitope

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非肌层浸润性膀胱癌激光整块切除研究进展

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微创泌尿外科杂志 2024年第2期13卷 140-143页

作者：王磊朱家红亳州市人民医院泌尿外科亳州236800

非肌层浸润性膀胱癌的标准治疗方式为经尿道膀胱肿瘤电切术(TURBT)辅以膀胱腔内化疗或免疫治疗。TURBT具有创伤小、恢复快、可反复切割等优点,但在安全性与病理精确评估方面存在一定的缺陷。而激光用于膀胱肿瘤时既不会产生闭孔神经反射... 详细信息

非肌层浸润性膀胱癌的标准治疗方式为经尿道膀胱肿瘤电切术(TURBT)辅以膀胱腔内化疗或免疫治疗。TURBT具有创伤小、恢复快、可反复切割等优点,但在安全性与病理精确评估方面存在一定的缺陷。而激光用于膀胱肿瘤时既不会产生闭孔神经反射,又可减少手术出血,目前开展的激光手术多以膀胱肿瘤整块切除术为主,激光因其汽化切割避免了组织炭化的可能,因此剜除的膀胱肿瘤基底部肌层组织得以完整保留,对病理的分级及临床分期有独到的优势。本文综述了激光膀胱肿瘤整块切除术的技术优势、现状与进展。

关键词：膀胱肿瘤激光整块切除

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SURFACE　REACTION，HYDROGEN　DIFFUSIVITY　AND　ENVIRONMENTAL　EMBRITTLEMENT　OF　INTERMETALLIC　COMPOUNDS　Ni_3Al　AND　Fe_3Al

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Acta Metallurgica Sinica(English Letters) 1995年第Z1期8卷 299-300,301302+303-3页

作者： WAN Xiaojing zhu jiahong HUANG Shengbiao(Shanghai University. Shanghai 200072, China) Shanghai Univ Shanghai China

The effect of boron doping on the sensitivity to environmental embrittlement of Ni3Al-based alloys was investigated in this paper. The results show that the ductilizing effect of boron in Ni3Al is partly to suppress moisture-induced hydrogen *** mechanism of this suppressing effect of boron relates to its severely decreasing the hydrogen diffusivity by boron segregated at the grain boundaries. The surface reaction of Fe3Al with water vapor and oxygen was experimentally confirmed by AES and XPS analysis. The kinetics of these reactions can be used to explain the different ductility behavior of aluminides in various environments.

关键词： environmental embrittlement hydrogen diffusivity surface reaction,intermetallic compound

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A simple nanobody-based competitive ELISA to detect antibodies against African swine fever virus

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Virologica Sinica 2022年第6期37卷 922-933页

作者： Jiakai Zhao jiahong zhu Ying Wang Mengting Yang Qiang Zhang Chong Zhang Yuchen Nan En-Min Zhou Yani Sun Qin Zhao Department of Preventive Veterinary Medicine College of Veterinary MedicineNorthwest A&F UniversityYangling Observing and Experimental Station of National Data Center of Animal HealthMinistry of AgricultureYangling712100China Kunming Customs Technology Center Kunming650228China

African swine fever virus(ASFV) infection is a big threat to the global pig industry. Because there is no effective vaccine, rapid, low-cost, and simple diagnosis methods are necessary to detect the ASFV infection in pig ***, with advantages of small molecular weight and easy genetic engineering, have been universally used as reagents for developing diagnostic kits. In this study, the recombinant ASFV-p30 was expressed and served as an antigen to immunize the Bactrian camel. Then, seven nanobodies against ASFV-p30 were screened using phage display technique. Subsequently, the seven nanobodies fused horseradish peroxidase(nanobody-HRP) were secretory expressed and one fusion protein ASFV-p30-Nb75-HRP was selected with the highest sensitivity in blocking ELISA. Using the ASFV-p30-Nb75-HRP fusion protein as a probe, a competitive ELISA(cELISA) was developed for detecting anti-ASFV antibodies in pig sera. The cut-off value of cELISA was determined to be 22.7%by testing 360 negative pig sera. The detection limit of the cELISA for positive pig sera was 1:320, and there was no cross-reaction with anti-other swine virus antibodies. The comparative assay showed that the agreement of the cELISA with a commercial ELISA kit was 100%. More importantly, the developed cELISA showed low cost and easy production as a commercial kit candidate. Collectively, a simple nanobody-based cELISA for detecting antibodies against ASFV is developed and it provides a new method for monitoring ASFV infection in the pig herds.

关键词： African swine fever virus(ASFV) Nanobody ASFV-p30 Nanobody-HRP fusion Protein Competitive ELISA

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Environmental Embrittlement in A_3B-type Intermetallic Alloys

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Journal of Materials Science & Technology 1994年第1期10卷 39-53页

作者： Xiaojing WAN jiahong zhu Kailiang JING Weijie CHEN and Yue WU(Institute of Metallurgy and Materials Science, Shanghai University of Technology, Shanghai, 200072, China) Shanghai Univ of Technology Shanghai China

Environmental embrittlement in A3B-type intermetallics based on Ni3Al and Fe3Al has been studied in this paper. For the Ni3Al doped with 120 wt ppm B and Ni,(Al,***) doped with 80 wt ppm B,their elongation and ultimate tensile strength decreased in the sequence:of vacuum > air >hydrogen. while for Ni,(Al,Mn) doped with 400 wt ppm B no envifonmental degradation was ob served, although a -Ni3(Al,Mn) alloy without B showed a decrease in ductility when tested in air in stead of oxygen. It is supposed that boron and hydrogen compete for the occupation of interstitial sites near grain boundaries. If boron content is sufficiently low, hydrogen embrittlement occurs ;however, if its content is sufficiently high. boron addition is capable of eliminating envjronmental ef fect in Ni3Al-based alloysi As to the micromechanism of hydrogen embrittlement in Ni3Al+B. S EM in situ observations showed that both grain boundary decohesion and a high stress concentration con tributed to hydrogen-assisted jntergranu lar cracking in this alloy. For the Fe3Al and Fe3 (***) *** mechanical properties depended strongly on grain size / grain shape and testing environment. A strain rate effect on ductiIity and fracture strength was also observed in the Fe3Al and Fe,(Al,Cr)+B aIloys. Preoxidation increased the ductility of the Fe,(Al,Cr)+B alloy. All these results can be rationalized from a hypothesis that surface reaction is the controlling process in embrittling Fe3Al-based alloys.

关键词： Cr type Environmental Embrittlement in A3B-type Intermetallic Alloys Figure

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贵州石漠化山区不同种植模式下的山豆根质量研究

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耕作与栽培 2024年第2期44卷 75-79页

作者：朱家红王小仲任得强龙菀成吴帆何艳利郭延红陈贇贵阳康养职业大学贵阳550081 关岭县中药材产业发展中心贵州关岭561300 贵州中医药大学贵阳550025

比较贵州石漠化山区不同种植模式下的山豆根药材质量,优选适宜的地区种植模式,对于产业发展及石漠化山区环境治理有较强现实意义。本研究设置仿野生、大田种植模式实验,以野生山豆根药材为对照,对山豆根苦参碱与氧化苦参碱含量、浸出物... 详细信息

比较贵州石漠化山区不同种植模式下的山豆根药材质量,优选适宜的地区种植模式,对于产业发展及石漠化山区环境治理有较强现实意义。本研究设置仿野生、大田种植模式实验,以野生山豆根药材为对照,对山豆根苦参碱与氧化苦参碱含量、浸出物含量、含水量、总灰分、重金属含量等性状测定和比较分析。结果表明,三种种植模式下,山豆根药材质量均符合《中华人民共和国药典》(2020年版)规定,其中仿野生种植模式下苦参碱和氧化苦参碱总含量为2.06%,浸出物含量22.77%,比大田种植模式、野生分别高14.75%、31.28%。仿野生栽培环境与大田栽培条件产量差异明显。研究表明,越南槐可采用仿野生种植模式在石漠化山区种植推广。

关键词：山豆根石漠化山区种植模式药材质量

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植物原生质体分离及其瞬时转化的应用

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热带生物学报 2024年第2期15卷 241-250页

作者：马超越彭世清郭冬朱家红王颖河北农业大学林学院河北保定071000 中国热带农业科学院热带生物技术研究所海口571101

原生质体已被广泛应用于亚细胞定位、基因表达分析、蛋白-蛋白互作、基因编辑等方面。笔者总结了酶解法分离植物原生质体的影响因素,电穿孔和PEG介导的方法对原生质体瞬时转化效率的影响和瞬时转化的应用。植物的种类、不同的PEG浓度、... 详细信息

原生质体已被广泛应用于亚细胞定位、基因表达分析、蛋白-蛋白互作、基因编辑等方面。笔者总结了酶解法分离植物原生质体的影响因素,电穿孔和PEG介导的方法对原生质体瞬时转化效率的影响和瞬时转化的应用。植物的种类、不同的PEG浓度、转染时间、DNA浓度和原生质体数量都会影响PEG介导法对原生质体瞬时转化效率。电穿孔法必须在脉冲电压、脉冲长度、脉冲数、细胞数、DNA浓度都适宜的条件下才能达到最佳转化效率。本文为更多植物原生质体分离体系和瞬时转化体系的建立提供参考。

关键词：原生质体酶解法瞬时转化

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用免疫金银染色法和免疫电镜观察重组痘苗病毒对狂犬病毒糖蛋白的表达

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中华实验和临床病毒学杂志 1994年第4期8卷 305-307页

作者：罗明朱家鸿卫生部武汉生物制品研究所

用负染及超薄切片的方法电镜观察了表达狂犬病毒糖蛋白的重组痘苗病毒ＴＴＶ１１ＫＲＧ株（简称重组病毒）的纯度、形态与结构，显示重组病毒呈典型的砖形及卵圆形。用特异的抗狂犬病毒糖蛋白的单克隆抗体为一抗，胶体金标记的羊抗鼠Ｉ... 详细信息

用负染及超薄切片的方法电镜观察了表达狂犬病毒糖蛋白的重组痘苗病毒ＴＴＶ１１ＫＲＧ株（简称重组病毒）的纯度、形态与结构，显示重组病毒呈典型的砖形及卵圆形。用特异的抗狂犬病毒糖蛋白的单克隆抗体为一抗，胶体金标记的羊抗鼠ＩｇＧ为二抗，作免疫金银染色，在普通光学显微镜下显示该重组病毒感染的ＣＶ－１细胞膜上存在大量金银颗粒的吸附。进一步作胶体金免疫电镜，显示该感染细胞经金标后细胞膜上存在大量电子密度致密的金颗粒，从而直观地证实了该重组病毒在感染的宿主细胞膜上有大量狂犬病毒糖蛋白的表达。

关键词：重组痘苗病毒狂犬病毒糖蛋白免疫金银染色

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Non-destructive buffer enabling near-infrared-transparent inverted inorganic perovskite solar cells toward 1400 h light-soaking stable perovskite/Cu（In,Ga）Se2 tandem solar cells

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Journal of Energy Chemistry 2024年第10期97卷 622-629页

作者： Yu Zhang Zhaoheng Tang Zhongyang Zhang jiahong Tang Minghua Li Siyuan zhu Wenyan Tan Xi Jin Tongsheng Chen Jinsong Hu Chao Zhou Chunlei Yang Qijie Liang Xugang Guo Weimin Li Weiqiang Chen Yan Jiang MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science Guangzhou Key Laboratory of Spectral Analysis and Functional Probes College of Biophotonics South China Normal University School of Materials Science and Engineering Beijing Institute of Technology Songshan Lake Materials Laboratory Department of Materials Science and Engineering Guangdong Provincial Key Laboratory of Functional Oxide Materials and Devices Southern University of Science and Technology Beijing Advanced Innovation Center for Soft Matter Science and Engineering College of Chemical Engineering College of Materials Science and Engineering Beijing University of Chemical Technology Department of Physics Liaoning University Beijing National Laboratory for Molecular Sciences (BNLMS) Institute of Chemistry Chinese Academy of Sciences School of Physics and Materials Science Guangzhou University Shenzhen Institute of Advanced Technology Chinese Academy of Sciences

Near-infrared（NIR） transparent inverted all-inorganic perovskite solar cells（PSCs） are excellent top cell candidates in tandem *** essential challenge is the replacement of metal contacts with transparent conductive oxide（TCO） electrodes,which requires the introduction of a buffer layer to prevent sputtering *** this study,we show that the conventional buffers（i.e.,small organic molecules and atomic layer deposited metal oxides） used for organic-inorganic hybrid perovskites are not applicable to all-inorganic perovskites,due to non-uniform coverage of the vulnerable layers underneath,deterioration upon ion bombardment and moisture induced perovskite phase transition,A thin film of metal oxide nanoparticles by the spin-coating method serves as a non-destructive buffer layer for inorganic ***-inorganic inverted near-infrared-transparent PSCs deliver a PCE of 17.46% and an average transmittance of 73.7% between 780 and 1200 *** combination with an 18.56% Cu（In,Ga）Se2bottom cell,we further demonstrate the first all-inorganic perovskite/CIGS 4-T tandem solar cell with a PCE of 24.75%,which exhibits excellent illumination stability by maintaining 86.7% of its initial efficiency after 1400 *** non-destructive buffer lays the foundation for efficient and stable NIR-transparent inverted inorganic perovskite solar cells and perovskite-based tandems.

关键词： Buffer layer CsPbI3 perovskite Inverted perovskite solar cells Stability Tandem solar cells

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