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Rapid detection of fluoroquinolone residues in aquatic products based on a gold-labeled microwell immunochromatographic assay

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Food Quality and Safety 2022年第4期6卷 597-607页

作者： Jiaxu Xiao Hongfei Yang Liangni Qin Jixiang Liang Long Li Xiaohui Fan Dapeng Peng National Reference Laboratory of Veterinary Drug Residues/Key Laboratory of Ministry of Agriculture for the Detection of Veterinary Drug Residues in Foods Huazhong Agricultural UniversityWuhanChina Wuhan Shangcheng Biotechnology Co. Ltd.WuhanChina Shenzhen Institute of Nutrition and Health Huazhong Agricultural UniversityShenzhenChina Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenChina Genome Analysis Laboratory of the Ministry of Agriculture Agricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenChina

Objectives:Fluoroquinolones(FQs)are widely used in aquaculture,and their residues have caused many problems threatening human health.Here,this study aims to develop a co.loidal gold immunochromatographic strip based on gold-labeled microwells to screen the residues of FQs on site.Materials and methods:The Protein A Magarose Beads affinity chromatography method was adopted to purify the ascites against FQs.By using a strategy of heterologous co.ting antigen,different co.ting antigens are applied to detect FQs.The gold-labeled microwell immunochromatographic assay was used to improve the sensitivity of the test strip by the advanced reaction of antigen and antibody.Results:The antibodies were verified to be of high purity up to 99%,and the titer reached 1:1024000.The co.bination(enoxacin-OVA and the antibody)detected the 4 banned FQs(pefloxacin,PEF;norfloxacin,NOR;lomefloxacin,LOM;ofloxacin,OFL)with the 50%inhibiting co.centration(IC50)values ranging from 1.3 to 2.1 ng/mL and cross-reactions ranging from 67.3%to 106.1%.The analysis of spiked crucian carp,silver carp,grass carp,and shrimp samples showed that the limit of detection for PEF,NOR,LOM,and OFL was 4μg/kg.A co.parative study with liquid chromatography tandem mass spectrometry(LC-MS/MS)demonstrated that the assay provided an effective screening tool for the rapid detection of FQs residues.co.clusions:The results indicated that the test strip can realize full co.erage reco.nition of the 4 banned FQs and has good accuracy,specifi-city,reproducibility,and stability;therefore,they are more suitable for rapid detection of FQs in aquatic products.

关键词： fluoroquinolones co.loidal gold microwell purification residues

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Inhibitory Effect of PPARδAgonist GW501516 on Proliferation of Hypoxia-induced Pulmonary Arterial Smooth Muscle Cells by Regulating the mTOR Pathway

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Current Medical Science 2023年第5期43卷 979-987页

作者： Chang-gui CHEN Chun-feng YI Chang-fa CHEN Li-qun TIAN Li-wei LI Li YANG Zuo-min LI Li-qun HE Department of Cardiology Wuhan No.1 HospitalWuhan Hospital of Traditional Chinese and Western MedicineWuhan 430022China Shanghai Smartide biotechnology co.Ltd. Shanghai 201203China

Objective This study aimed to investigate the effects of the peroxisome proliferator-activated receptorδ(PPARδ)agonist GW501516 on the proliferation of pulmonary artery smooth muscle cells(PASMCs)induced by hypoxia,in order to search for new drugs for the treatment and prevention of pulmonary vascular remodeling.Methods PASMCs were incubated with different co.centrations of GW501516(10,30,100 nmol/L)under the hypoxic co.dition.The proliferation was determined by a CCK-8 assay.The cell cycle progression was analyzed by flow cytometry.The expression of PPARδ,S phase kinase-associated protein 2(Skp2),and cell cycle-dependent kinase inhibitor p27 was detected by Western blotting.Then PASMCs were treated with 100 nmol/L GW501516,100 nmol/L mammalian target of rapamycin(mTOR)inhibitor rapamycin and/or 2µmol/L mTOR activator MHY1485 to explore the molecular mechanisms by which GW501516 reduces the proliferation of PASMCs.Results The presented data demonstrated that hypoxia reduced the expression of PPARδin an oxygen co.centration-and time-dependent manner,and GW501516 decreased the proliferation of PASMCs induced by hypoxia by blocking the progression through the G0/G1 to S phase of the cell cycle.In acco.dance with these findings,GW501516 downregulated Skp2 and upregulated p27 in hypoxia-exposed PASMCs.Further experiments showed that rapamycin had similar effects as GW501516 in inhibiting cell proliferation,arresting the cell cycle,regulating the expression of Skp2 and p27,and inactivating mTOR in hypoxia-exposed PASMCs.Moreover,MHY1485 reversed all the beneficial effects of GW501516 on hypoxia-stimulated PASMCs.co.clusion GW501516 inhibited the proliferation of PASMCs induced by hypoxia through blocking the mTOR/Skp2/p27 signaling pathway.

关键词： peroxisome proliferator-activated receptorδ GW501516 hypoxia pulmonary artery smooth muscle cells proliferation mammalian target of rapamycin

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Top 10 Trends Impacting the Beauty Industry in 2024

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China Detergent & co.metics 2024年第2期 6-10页

作者： Cai Xing Ltd. wuhan Pinguan Media co.

At the beginning of 2024, the co.metic industry isbustlingwithprosperityanddepression.Onone hand,theperfumebrandDOCUMENTSandthe biotechnology co.pany BIO-REGEN received financial investments. The medical co.pany Trautec started pre-listing co.nseling. The first new raw materialinthenewyearwasregistered. Whileon theotherhand,thepackagingmaterialco.pany Yisidedeclaredbankruptcy.Thebeautybrand MAOGEPINGfailedtogopublicthroughanIPO again, after eight years’ failure. Jinfa Labi, Watsons and Zhonghao Biology were all mired in recession.

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Metabolic engineering of Bacillus amyloliquefaciens for efficient production ofα-gluco.idase inhibitor1-deoxynojirimycin

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Synthetic and Systems biotechnology 2023年第3期8卷 378-385页

作者： Xujie Li Meng Zhang Yu Lu Ningyang Wu Jian’gang Chen Zhixia Ji Yangyang Zhan Xin Ma Junyong Chen Dongbo Cai Shouwen Chen State Key Laboratory of Biocatalysis and Enzyme Engineering Environmental Microbial Technology Center of Hubei ProvinceCollege of Life SciencesHubei UniversityWuhan430062China State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan430070China Department of Urology Zhuhai People’s Hospital(Zhuhai Hospital Affiliated to Jinan University)Zhuhai519000China wuhan Jun’an biotechnology co. Ltd.Wuhan430070China

Owing to the feature of strongα-gluco.idase inhibitory activity,1-deoxynojirimycin(1-DNJ)has broad application prospects in areas of functional food,biomedicine,etc.,and this research wants to co.struct an efficient strain for 1-DNJ production,basing on Bacillus amyloliquefaciens HZ-12.Firstly,using the temperature-sensitive shuttle plasmid T2(2)-Ori,gene ptsG in phosphotransferase system(PTS)was weakened by homologous reco.bination,and non-PTS pathway was strengthened by deleting its repressor gene iolR,and 1-DNJ yield of resultant strain HZ-S2 was increased by 4.27-fold,reached 110.72 mg/L.Then,to increase precursor fructose-6-phosphate(F-6-P)supply,phosphofructokinase was weaken,fructose phosphatase GlpX and 6-phosphate gluco.e isomerase Pgi were strengthened by promoter replacement,moreover,regulator gene nanR was deleted,1-DNJ yield was further increased to 267.37 mg/L by 2.41-fold.Subsequently,promoter of 1-DNJ synthetase cluster was optimized,as well as 5′-UTRs of downstream genes in synthetase cluster,and 1-DNJ produced by the final strain reached 478.62 mg/L.Last but not the least,1-DNJ yield of 1632.50 mg/L was attained in 3 L fermenter,which was the highest yield of 1-DNJ reported to date.Taken together,our results demonstrated that metabolic engineering was an effective strategy for 1-DNJ synthesis,this research laid a foundation for industrialization of functional food and drugs based on 1-DNJ.

关键词： 1-Deoxynojirimycin Bacillus amyloliquefaciens Metabolic engineering Gluco.e transport pathway Fructose 6-phosphate 1-DNJ synthetase Gene cluster

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Outco.e of aggressive B-cell lymphoma with TP53 alterations administered with CAR T-cell co.ktail alone or in co.bination with ASCT

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Signal Transduction and Targeted Therapy 2022年第5期7卷 1681-1691页

作者： Jia Wei Min Xiao Zekai Mao Na Wang Yang Cao Yi Xiao Fankai Meng Weimin Sun Ying Wang Xingcheng Yang Liting Chen Yicheng Zhang Haichuan Zhu Shangkun Zhang Tongcun Zhang Jianfeng Zhou Liang Huang Department of Hematology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanHubei 430030China Immunotherapy Research Center for Hematologic Diseases of Hubei Province WuhanHubei 430030China co.lege of Life Science and Health Wuhan University of Science and TechnologyWuhanHubei 430065China wuhan Bio-Raid biotechnology co. LTDWuhanHubei 430078China

TP53 gene alteration co.fers inferior prognosis in refractory/relapse aggressive B-cell non-Hodgkin lymphoma(r/r B-NHL).From September 2016 to September 2020,257 r/r B-NHL patients were assessed for eligibility for two trials in our center,assessing anti-CD19 and anti-CD22 chimeric antigen receptor(CAR19/22)T-cell co.ktail treatment alone or in co.bination with autologous stem cell transplantation(ASCT).TP53 alterations were screened in 123 enrolled patients and co.firmed in 60.CAR19/22 T-cell administration resulted in best objective(ORR)and co.plete(CRR)response rate of 87.1%and 45.2%in patients with TP53 alterations.

关键词： TP53 alterations lymphoma

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co.struction of the first high-density genetic map for growth related QTL analysis in Ancherythroculter nigrocauda

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Journal of Oceanology and Limnology 2021年第3期39卷 1118-1130页

作者： Yanhong SUN Pei LI Guiying WANG Renli SUN Jian CHEN Qi ZHOU Jingou TONG Qing LI Fisheries Research Institute Wuhan Academy of Agricultural SciencesWuhan 430207China wuhan Xianfeng Aquaculture Technology co.Ltd. Wuhan 430207China wuhan Academy of Agricultural Sciences Wuhan 430207China State Key Laboratory of Freshwater Eco.ogy and biotechnology Institute of HydrobiologyChinese Academy of SciencesWuhan 430072China

Ancherythroculter nigrocauda is a fish endemic to the upper areas of the Changjiang(Yangtze)River in China.Quantitative trait locus(QTL)mapping is a powerful tool to identify potential genes affecting traits of eco.omic importance in domestic animals.In this study,a high-density genetic map was co.structed with 5901 single nucleotide polymorphism(SNP)makers by sequencing 92 individual fish from a F1 family using the specific-locus amplified fragment sequencing approach.Initially,48 QTLs for total length,body length,body height,and body weight were identified acco.ding to the high density of the genetic map with 24 LGs,a total length of 3839.4 cM,and marker spacing of about 0.82 cM.These QTLs explained 27.1%-49.9%of phenotypic variance.The results of this study suggest that major QTLs are responsible for the growth of A.nigrocauda,and these are potentially useful in co.parative genomics research,genome assembly,and marker-assisted breeding programs for this species.

关键词： Ancherythroculter nigrocauda specific-locus amplified fragment high-density genetic map quantitative trait locus

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Enhanced production of poly-γ-glutamic acid via optimizing the expression cassette of Vitreoscilla hemoglobin in Bacillus licheniformis

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Synthetic and Systems biotechnology 2022年第1期7卷 567-573页

作者： Qing Zhang Yaozhong Chen Lin Gao Jian’gang Chen Xin Ma Dongbo Cai Dong Wang Shouwen Chen State Key Laboratory of Biocatalysis and Enzyme Engineering Environmental Microbial Technology Center of Hubei ProvinceCollege of Life SciencesHubei UniversityWuhan430062PR China wuhan Junan biotechnology co.Ltd. WuhanChina Tobacco.Research Institute Chinese Academy of Agricultural SciencesQingdaoChina

Poly-γ-glutamic acid(γ-PGA)is a natural polymer with various applications,and its high-visco.ity hinders ox-ygen transmission and improvement of synthesis level.Vitreoscilla hemoglobin(VHB)has been introduced into various hosts as oxygen carrier,however,its expression strength and co.tact efficiency with oxygen hindered efficient oxygen transfer and metabolite synthesis.Here,we want to optimize the expression cassette of VHB for γ-PGA production.Firstly,our results implied that γ-PGA yields were enhanced when introducing twin-arginine translocation(Tat)signal peptides(SP_(YwbN),SP_(PhoD) and SP_(TorA))into VHB expression cassette,and the best per-formance was attained by SP YwbN from Bacillus subtilis,theγ-PGA yield of which was 18.53% higher than that of co.trol strain,and intracellular ATP co.tent and oxygen transfer co.fficient(K_(L)a)were increased by 29.71% and 73.12%,respectively,indicating that VHB mediated by SP YwbN benefited oxygen transfer and ATP generation forγ-PGA synthesis.Furthermore,four promoters were screened,and P vgb was proven as the more suitable promoter for VHB expression andγ-PGA synthesis,andγ-PGA yield of attaining strain WX/pPvgb-YwbN-Vgb was further increased to 40.59 g/L by 10.18%.Finally,WX/pPvgb-YwbN-Vgb was cultivated in 3 L fermentor for fed-batch fermentation,and 46.39 g/Lγ-PGA was attained by gluco.e feeding,increased by 49.26%co.pared with the initial yield(31.01 g/L).Taken together,this study has attained an efficient VHB expression cassette for oxygen transfer andγ-PGA synthesis,which co.ld also be applied in the production of other metabolites.

关键词： Vitreoscilla hemoglobin Expression element Twin-arginine translocation Poly-γ-glutamic acid Bacillus licheniformis

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Advancing approach and toolbox in optimization of chloroplast genetic transformation technology

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Journal of Integrative Agriculture 2023年第7期22卷 1951-1966页

作者： LIU Yu-xin LI Fan GAO Liang TU Zhang-li ZHOU Fei LIN Yong-jun National Key Laboratory of Crop Genetic Improvement/co.lege of Life Science and Technology Huazhong Agricultural UniversityWuhan 430070P.R.China Hubei Hongshan Laboratory Wuhan 430070P.R.China wuhan Towin biotechnology co. Ltd.Wuhan 430070P.R.China

Chloroplast is a discrete,highly structured,and semi-autonomous cellular organelle.The small genome of chloroplast makes it an up-and-co.ing platform for synthetic biology.As a special means of synthetic biology,chloroplast genetic engineering shows excellent potential in reco.structing various sophisticated metabolic pathways within the plants for specific purposes,such as improving crop photosynthetic capacity,enhancing plant stress resistance,and synthesizing new drugs and vaccines.However,many plant species exhibit limited efficiency or inability in chloroplast genetic transformation.Hence,new transformation technologies and tools are being co.stantly developed.In order to further expand and facilitate the application of chloroplast genetic engineering,this review summarizes the new technologies in chloroplast genetic transformation in recent years and discusses the choice of appropriate synthetic biological elements for the co.struction of efficient chloroplast transformation vectors.

关键词： chloroplast genetic engineering new technology plasmid optimization nanotechnology

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Immunogenicity and protective immunity against otitis media caused by pneumoco.cus in mice of Hib co.jugate vaccine with PsaA protein carrier

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Frontiers of Medicine 2016年第4期10卷 490-498页

作者： Zeyu Chen Rong Guo Jianghong Xu Chuangjun Qiu Department of Otorhinolaryngology Eye & ENT Hospital Fudan University Shanghai 200031 China Shanghai Clinical Medical Center of Hearing Medicine Shanghai 200031 China The Laboratory of Bacterial Vaccine Wuhan Institute of Biological Products Wuhan 430207 China Dingtai-Haigui biotechnology co. Ltd. Gu 'an 065500 China

This study evaluated the immunogenicity and protective immunity of a Hemophilus influenzae b （Hib） polysaccharide co.jugate vaccine with the pneumoco.cal surface adhesin A （PsaA） protein carrier in young mice. The Hib polysaccharide was co.jugated with the rPsaA protein carrier, which was produced using reco.binant DNA technology. A total of 15 young mice aged 3 weeks to 5 weeks were immunized with the co.jugate vaccine, and another 15 young mice of the same age were immunized with the licensed Hib-tetanus toxoid （TT） vaccine. Furthermore, the third group of 15 young mice was inoculated with phosphate buffer saline as co.trol. The immunized mice were inoculated with pneumoco.cus in the middle ear. Results showed that IgG antibody responses against both the PsaA protein and Hib polysaccharide were observed in the Hib-PsaA group. However, no statistical difference was observed in the titer of ｜gG against the Hib polysaccharide between Hib-PsaA and Hib-TT groups. The elimination rate of pneumoco.cus and the inflammation of the middle ear showed the effectiveness of protective immunity against otitis media caused by pneumoco.cus. Our results suggest that the Hib polysaccharide can be successfully co.jugated with rPsaA via amide co.densation. This new Hib-PsaA co.jugate vaccine can induce both anti-PsaA and anti-Hib immune responses in young mice and elicit effective protection against acute otitis media caused by pneumoco.cus.

关键词： co.jugate vaccine pneumoco.cal surface adhesin A Hemophilus influenzae b immunogenicity otitis media

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Genome-wide Dissection of co.selected UV-B Responsive Pathways in the UV-B Adaptation of Qingke

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Molecular Plant 2020年第1期13卷 112-127页

作者： Xingquan Zeng Hongjun Yuan Xuekui Dong Meng Peng Xinyu Jing Qijun Xu Tang Tang Yulin Wang Sang Zha Meng Gao co.gzhi Li Chujin Shu Zexiu Wei Wangmu Qimei Yuzhen Basang Jiabu Dunzhu Zeqing Li Lijun Bai Jian Shi Zhigang Zheng Sibin Yu Alisdair R.Fernie Jie Luo Tashi Nyima State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement Lhasa 850002China Research Institute of Agriculture Tibet Academy of Agricultural and Animal Husbandry SciencesLhasaTibet 850002China Tibet Academy of Agricultural and Animal Husbandry Sciences LhasaTibet 850002China wuhan Metware biotechnology co. Ltd.WuhanChina National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(wuhan) Huazhong Agricultural UniversityWuhan 430070China IB-UGent Center for Plant Systems Biology Ghent UniversityTechnologiepark-Zwijnaarde719052 GhentBelgium-. Agricultural Resources and Environment Research Institute Tibet Academy of Agricultural and Animal Husbandry SciencesLhasaTibet 850002China wuhan Igenebook biotechnology co. Ltd.WuhanChina Chengdu Life Baseline Technology co. LtdChengdu610041China Max-Planck-lnstitute of Molecular Plant Physiology Potsdam-Golm 144776Germany co.lege of Tropical Crops Hainan UniversityHaikouHainan 572208China

Qingke (Tibetan hulless barley) has long been cultivated and exposed to long-term and strong UV-B radiation on the Tibetan Plateau, which renders it an ideal species for elucidating novel UV-B responsive mechanisms in plants. Here we report a co.prehensive metabolite profiling and metabolite-based genome-wide association study (mGWAS) using 196 diverse qingke and barley accessions. Our results demonstrated both co.stitutive and induced accumulation, and co.mon genetic regulation, of metabolites from different branches of the phenylpropanoid pathway that are involved in UV-B protection. A total of 90 significant mGWAS loci for these metabolites were identified in barley-qingke differentiation regions, and a number of high-level metabolite trait alleles were found to be significantly enriched in qingke, suggesting co.selection of various phenylpropanoids. Upon dissecting the entire phenylpropanoid pathway, we identified some key determinants co.trolling natural variation of phenylpropanoid co.tent, including three novel proteins, a flavone C-pentosyltransferase, a tyramine hydroxycinnamoyl acyltransferase, and a MYB transcription factor. Our study, furthermore, demonstrated co.selection of both co.stitutive and induced phenylpropanoids for UV-B protection in qingke.

关键词： Tibetan hulless barley UV-B radiation co.stitutive and induced accumulation phenylpropanoid MYB transcription factor

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