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Combination of iron flocculation and qPCR for quantitative evaluation of virus-shedding intensity of goldfish Carassius auratus infected with cyprinid herpesvirus 2 in the water and the effect of sodium chlorite powder in blocking waterborne horizontal viral transmission

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Aquaculture and Fisheries 2024年第4期9卷 617-625页

作者： Yufei Zhai Fei Shen Hao Wang liqun lu National Pathogen Collection Center for Aquatic Animals Shanghai Ocean UniversityShanghai201306China Key Laboratory for Freshwater Aquatic Genetic Resources at Shanghai Ocean University Ministry of Agriculture and Rural AffairsShanghai201306China College of Fisheries and Life Science Shanghai Ocean UniversityShanghai201306China

Cyprinid herpesvirus 2(CyHV-2),a member of the Alloherpesviridae family belonging to the genus Cyprinivirus,was initially isolated from goldfish(Carassius auratus)and has been recently emerging as a virulent pathogen for cultured prussian carp(Carassius auratus gibelio)*** this study,a novel and effective method for concentration and quantification of live CyHV-2 virions from water was successfully established through coupling the iron flocculation with real time qPCR ***,the shedding intensity of CyHV-2 in fish-tank water from artificially-challenged goldfish(25 fish/20 L)was monitored continuously for 7 days on a daily basis through quantitating viral genomic copy numbers by qPCR,and the maximum shedding level was determined to be 105 copies/*** transmission research system was established by inoculating healthy goldfish in water spiked with serial dilution of CyHV-2 virions ranging from 107 to 103 copies/*** results indicated that water-borne CyHV-2 efficiently caused the infection of tested goldfish even in a concentration of 103 copies/L,and the overall transmission efficacy was not linearly correlated with the level of input virus in the fish *** disinfectant Composite Sodium Chlorite Powder(CSCP)has been widely applied in aquaculture to control microbial infection through direct spill in the water,and its effect in inactivating the CyHV-2 infectivity remains *** further determined that the EC50 of CSCP against 3.89 TCID50/mL CyHV-2 was close to 15.625μg/mL in vitro,and application of CSCP in a level as high as 60μg/mL(the safety concentration of CSCP for goldfish)couldn’t protect goldfish from CyHV-2 challenge through ***,the disinfectant CSCP was regarded as none-effective for blocking CyHV-2 transmission in water during ***,our data provided quantitative data to demonstrate the shedding intensity of CyHV-2 in water,and CSCP was shown to be not effective in blocking water-borne horizontal

关键词： Cyprinid herpesvirus 2 Iron flocculation Virus shedding Water-borne horizontal transmission Composite sodium chlorite powder

MicroRNA-145-3p suppresses the malignant behaviors of T-cell acute lymphoblastic leukemia Jurkat cells via inhibiting the NF-kappaB signaling pathway

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BIOCELL 2020年第1期44卷 101-110页

作者： Xin YANG liqun lu Li HUANG Jing HE Jie LV Department of Pediatrics The First Affiliated Hospital of Chengdu Medical CollegeChengduChina

T-cell acute lymphoblastic leukemia(T-ALL)is a hematological tumor caused by the malignant transformation of immature T-cell progenitor *** studies have stated that microRNAs(miRNAs)may play key roles in T-ALL *** study aimed to investigate the roles of miR-145-3p in T-ALL cell proliferation,invasion,and apoptosis with the involvement of the nuclear factor-kappaB(NF-κB)signaling pathway.T-ALL Jurkat cells were harvested,and the expression of miR-145-3p and NF-κB-p65 was ***-and loss-of-functions of miR-145-3p and NF-κB-p65 were performed to identify their roles in the biological behaviors of Jurkat cells,including proliferation,apoptosis,and ***,the current study demonstrated that miR-145-3p was down-regulated while NF-κB-p65 was up-regulated in Jurkat ***-145-3p directly bound to the 3’untranslated region of NF-κ***-expression of miR-145-3p inhibited Jurkat cell proliferation,invasion,and resistance to apoptosis,while over-expression of NF-κB-p65 presented opposite ***-transfection of miR-145-3p and NF-κB-p65 promoted the malignant behaviors of Jurkat cells compared to miR-145-3p transfection alone,while it reduced these behaviors of Jurkat cells compared to NF-κB-p65 transfection *** together,this study provided evidence that miR-145-3p could suppress proliferation,invasion,and resistance to the death of T-ALL cells via inactivating the NF-κB signaling pathway.

关键词： T-cell acute lymphoblastic leukemia Proliferation Invasion Resistance to apoptosis

New method for cotton fractional vegetation cover extraction based on UAV RGB images

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International Journal of Agricultural and Biological Engineering 2022年第4期15卷 172-180页

作者： Huanbo Yang Yubin Lan liqun lu Daocai Gong Jianchi Miao Jing Zhao School of Agricultural and Food Engineering Shandong University of TechnologyZibo 255000ShandongChina International Precision Agriculture Aviation Application Technology Research Center Shandong University of TechnologyZibo 255000ShandongChina School of Transportation and Vehicle Engineering Shandong University of TechnologyZibo 255000ShandongChina

As the key principle of precision farming,the distribution of fractional vegetation cover is the basis of crop management within the field *** estimate crop FVC rapidly at the farm scale,high temporal-spatial resolution imagery obtained by unmanned aerial vehicle(UAV)was *** verify the application potential of consumer-grade UAV RGB imagery in estimated FVC,blue-green characteristic vegetation index(TBVI)and red-green vegetation index(TRVI)were proposed in this study according to the differences of the gray value among cotton vegetation,soil and shadow in the ***,two new constructed indices and several published indices were used to extract visible light images and generate greyscale images for each of the visible light vegetation ***,the thresholds of cotton vegetation and non-vegetation pixels were established based on the vegetation index threshold method which combines support vector machine classification and vegetation ***,the accuracy difference in vegetation information extraction between the newly constructed and several published indices was *** results show that the accuracy of the information extracted by TRVI is higher than that of subdivision index of other visible light(FVC extraction precision in the first bud stage of cotton:R2=0.832,RMSE=2.307,nRMSE=4.405%;FVC extraction precision in the bud stage of cotton:R2=0.981,RMSE=1.393,nRMSE=1.984%;FVC extraction precision in the flowering stage of cotton:R2=0.893,RMSE=2.101,nRMSE=2.422%;FVC extraction precision in the boll stage of cotton:R2=0.958,RMSE=1.850,nRMSE=2.050%).

关键词： cotton UAV visible light images fractional vegetation cover vegetation index threshold method TRVI TBVI

Development of a Fast ELISA for the Specific Detection of both Leucomalachite Green and Malachite Green

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Journal of Ocean University of China 2015年第2期14卷 340-344页

作者： JIANG Yousheng CHEN Li HU Kun YU Wenjuan YANG Xianle lu liqun College of Fisheries and Life Science Shanghai Ocean University

Malachite green(MG), a dye, is an antifungal agent that has been used to treat and prevent fish diseases. It is metabolized into reduced leucomalachite green forms(LMG) that may reside in fish muscles for a long period, thus being harmful to human health. The aim of this study was to develop a competitive and direct enzyme-linked immunosorbent assay(ELISA) to detect MG and LMG specifically. The monoclonal antibody(m Ab) to LMG was generated using a hybridoma technique. The obtained m Ab showed good cross-reactivity(CR) to malachite green(MG), but not to crystal violet(CV) and Brilliant Green(BG). The m Ab was used to develop a fast detecting ELISA of MG and LMG in fish. By introducing the conjugation LMG-HRP, the detection capability was 0.37 ng m L-1 for MG and LMG. The mean recovery from spiked grass carp tissues ranged from 76.2% to 82.9% and the coefficients of variation varied between 1.8% and 7.5%. The stable and efficient monoclonal cell line obtained is a sustainable source of sensitive and specific antibody to MG and LMG.

关键词： monoclonal antibody Malachite Green ELISA fish

Detection of cotton waterlogging stress based on hyperspectral images and convolutional neural network

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International Journal of Agricultural and Biological Engineering 2021年第2期14卷 167-174页

作者： Jing Zhao Fangjiang Pan Zhiming Li Yubin Lan liqun lu Dongjian Yang Yuting Wen School of Agricultural Engineering and Food Science Shandong University of TechnologyZibo 255000ShandongChina Sub-center of National Center for International Collaboration Research on Precision Agricultural Aviation Pesticides Spraying Technology Shandong University of TechnologyZibo 255000ShandongChina School of Traffic and vehicle engineering Shandong University of TechnologyZibo 255000ShandongChina

Waterlogging in the early stage of cotton will reduce the number of bolls and do harm to *** detection of waterlogging will help farmers to adjust cotton management and save the *** evaluate the application of deep learning for the detection of early waterlogging,this study applied a convolutional neural network(CNN)to classify different durations of waterlogging stress(0,2,4,6,8,10 d)based on hyperspectral images(HSIs)of cotton *** experiment was designed to simulate the situation of cotton under waterlogging stress and collect HSIs of visible and near-infrared(VNIR 450-950 nm)spectra with 126 bands 66 d after cotton sowing(66 DAS).It was found the spectral curve reflectance of waterlogging cotton was higher than that of non-waterlogging *** near 550 nm and 750 nm,and the spectral curve increased with durations of waterlogging stress and there were‘blue shift’phenomena for the position of the red edge of the *** first principal components of HSIs after band randomly discarding and principal component analysis(PCA)were used to build a *** Inception-v3(GLNI-v3)and VGG-16 models were selected to detect cotton waterlogging stress with the *** results showed that the average time for a round training for GLNI-v3 was 13.337 s,with a classification accuracy of 96.95%and a loss value of *** average time for a round training for VGG-16 was 21.470 s,with a classification accuracy of 97.00%and a loss value of *** these two models had similar classification accuracy and loss value,GLNI-v3 achieved a high accuracy with fewer training *** durations of waterlogging stress of cotton in a short-term can be detected by HSIs of cotton leaves and CNN models are suitable for the classification of HSIs,and this method can provide support for cotton yield estimation and loss assessment after waterlogging.

关键词： cotton waterlogging hyperspectral image convolutional neural network

Characterization of grass carp FosB, Fosl2, JunD transcription factors in response to GCRV infection

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Aquaculture and Fisheries 2022年第3期7卷 304-312页

作者： Fei Yu Wanjuan Li Longlong Wang Shunzheng Que liqun lu National Pathogen Collection Center for Aquatic Animals Shanghai Ocean UniversityShanghai201306China Institute of Marine Biology College of OceanographyHohai UniversityNanjing210098China Key Laboratory of Agriculture Ministry for Freshwater Aquatic Genetic Resources Shanghai Ocean UniversityShanghai201306China National Experimental Teaching Demonstration Center for Fishery Sciences Shanghai Ocean UniversityShanghai201306China

Activating protein-1(AP-1)composed of Jun and Fos family proteins,which are frequently activated by pathogens to induce cellular inflammation and immune *** this study,by using RNA sequencing(RNA-seq)technology,we found that FosB,Fosl2,and JunD expression was co-induced 12 and 20 h in grass carp(Ctenopharyngodon idella)kidney cell line(CIK cells)after grass carp reovirus(GCRV)***-length cDNA of the three genes were cloned and phylogenetic analysis indicated that the corresponding proteins shared high homology with those in higher *** tissue distribution analysis of the three genes showed they were ubiquitously expressed in various tissues of healthy grass *** infection assay,FosB,Fosl2,and JunD were up-regulated and co-expressed 8 h after GCRV infection in CIK *** interaction and co-expression network analysis revealed that FosB,Fosl2,and JunD showed similar functions and were co-expressed in mammals and *** together,these results indicated that the co-expression of the AP-1 family proteins in grass carp uniformly responds to GCRV *** findings provided insights into the further study of AP-1 transcription factors function during grass carp reovirus infection.

关键词： Grass carp reovirus FosB Fosl2 JunD Co-expression Inflammation

In vivo effects of neomycin sulfate on non-specific immunity,oxidative damage and replication of cyprinid herpesvirus 2 in crucian carp(Carassius auratus gibelio)

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Aquaculture and Fisheries 2019年第2期4卷 67-73页

作者： Penghui Sun Fei Yu Jianfei lu Minli Zhang Hao Wang Dan Xu liqun lu National Pathogen Collection Center for Aquatic Animals Shanghai Ocean UniversityShanghai 201306China Key Laboratory of Agriculture Ministry for Freshwater Aquatic Genetic Resources Shanghai Ocean UniversityShanghai 201306China

Neomycin belongs to the family of 2-deoxystreptamine-containing aminoglycoside *** is widely used for bacterial infections,targeting most gram-negative and some gram-positive *** has also been reported to show antiviral ***,we evaluated the toxicity of neomycin sulfate,and investigated its effect on non-specific immunity and viral infection in crucian carp(Carassius auratus gibelio).The safe concentration of neomycin sulfate for crucian carp was determined to be 102.9 mg/kg in *** oxidative damage assays,neomycin sulfate increased superoxide dismutase and catalase activity and decreased malondialdehyde in the liver of crucian *** non-specific blood immune assays,the white blood cell count and complement 3 content significantly increased after neomycin sulfate treatment,while no significant changes were observed in antibacterial or lysozyme *** a challenge test,neomycin sulfate protected the crucian carp from cyprinid herpesvirus 2(CyHV-2)infection and inhibited CyHV-2 *** cytotoxicity assays,low concentrations of neomycin sulfate had no cytotoxicity on cells from the fins of crucian *** results of the present study indicate that oral administration of neomycin sulfate reduced oxidative damage,enhanced immunity and provided protection against CyHV-2 in crucian carp.

关键词： Neomycin sulfate Non-specific immunity Oxidative damage Crucian carp Cyprinid herpesvirus 2

Identification of serum immunoglobulin M(IgM)of crucian carp(Carassius auratus gibelio)and immune response to cyprinid herpesvirus 2 infection

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Aquaculture and Fisheries 2024年第2期9卷 172-180页

作者： Shuran Wang Haotian Wu Longlong Wang Pinhua Wang liqun lu Yousheng Jiang Ye Zhang Dan Xu National Pathogen Collection Center for Aquatic Animals Shanghai Ocean UniversityShanghai201306China Key Laboratory of Aquatic Genetic Resources of the Ministry of Agriculture Shanghai Ocean UniversityShanghai201306China National Demonstration Center for Experimental Fisheries Science Education Shanghai Ocean UniversityShanghai201306China

Crucian carp(Carassius auratus gibelio),an extensively cultivated freshwater fish,was one of the model species for the study of fish *** antibodies were advantageous molecular tools for studying teleost immune ***,polyclonal antibodies reacting with immunoglobulins(Ig)were used successfully in studies of the teleost *** the present study,we produced polyclonal antibody against CH2 domains of crucian carp IgM,and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 *** recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic *** specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp *** RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 *** expression of IgM in the kidney increased and reached a maximum at 6 h post-infection(hpi),while dropped to a low level at 5 days post-infection(dpi).In conclusion,the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2,indicating that IgM played a potential role in systemic immunity against viral *** antibody against crucian carp IgM had certain clinical relevance,which might provide insight into the early stage of virus infection and prevention of the disease.

关键词： Immunoglobulin M Crucian carp Polyclonal antibody Cyprinid herpesvirus 2 Immunoprotection

Regulated emission characteristics of in-use LNG and diesel semi-trailer towing vehicles under real driving conditions using PEMS

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Journal of Environmental Sciences 2020年第2期32卷 155-164页

作者： liqun Lv Yunshan Ge Zhe Ji Jianwei Tan Xin Wang Lijun Hao Zhiwei Wang Mengzhu Zhang Chunjie Wang Haidan Liu National Laboratory of Auto Performance&Emission Test School of Mechanical EngineeringBeijing Institute of TechnologyBeijing 100081China Vehicle Emission Control Center Chinese Research Academy of Environmental SciencesBeijing 100081China Tangshan Environmental Monitoring Centre Tangshan 063000China

On-road driving emissions of six liquefied natural gas(LNG) and diesel semi-trailer towing vehicles(STTVs) which met China Emission Standard IV and V were tested using Portable Emission Measurement System(PEMS) in northern *** characteristics of these vehicles under real driving conditions were analyzed and proved that on-road emissions of heavy-duty vehicles(HDVs) were underestimated in the *** were large differences among LNG and diesel vehicles, which also existed between China V vehicles and China IV *** factors showed the highest level under real driving conditions, which probably be caused by frequent acceleration, deceleration, and *** emission factors ranged from 2.855 to 20.939 g/km based on distance-traveled and 6.719–90.557 g/kg based on fuel consumption during whole tests, which were much higher than previous researches on chassis *** was inferred from tests that the fuel consumption rate of the test vehicles had a strong correlation with NOx emission, and the exhaust temperature also affected the efficiency of Selected Catalytic Reduction(SCR) aftertreatment system, thus changing the NOx emission *** emission factors of LNG vehicles were 2.012–10.636 g/km, which were much higher than that of diesel vehicles(0.029–0.185 g/km).Unburned CH4 may be an important reason for this *** on-road emission tests, especially CH4 emission test should be carried out in subsequent *** addition, the Particulate Number(PN) emission factors of diesel vehicles were at a very high level during whole tests, and Diesel Particulate Filter(DPF)should be installed to reduce PN emission.

关键词： Semi-trailer towing vehicles Regulated emission PEMS