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Pig macrophages with site-specific edited CD163 decrease the susceptibility to infection with porcine reproductive and respiratory syndrome virus

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Journal of Integrative Agriculture 2023年第7期22卷 2188-2199页

作者： xu Kui ZHOU Yan-rong SHANG Hai-tao xu chang-jiang TAO Ran HAO Wan-jun LIU Sha-sha MU Yu-lian XIAO Shao-bo LI Kui State Key Laboratory of Animal Nutrition/Key Laboratory of Animal Genetics Breeding and Reproduction of Ministry of Agriculture and Rural AffairsInstitute of Animal SciencesChinese Academy of Agricultural SciencesBeijing 100193P.R.China Shenzhen Kingsino Technology Co. Ltd.Shenzhen 518106P.R.China State Key Laboratory of Agricultural Microbiology/Key Laboratory of Preventive Veterinary Medicine in Hubei Province College of Veterinary MedicineHuazhong Agricultural UniversityWuhan 430070P.R.China Precision Medicine Institute The First Affiliated HospitalSun Yat-sen UniversityGuangzhou 510080P.R.China

Porcine reproductive and respiratory syndrome(PRRS)is recognized as one of the most infectious viral diseases of swine.Although Cluster of differentiation 163(CD163)is identified as an essential receptor for mediating PRRS virus(PRRSV)infection,the important residues involved in infection on CD163 are still unclear.Therefore,it is very important to identify these key residues to study the mechanism of PRRSV infection and to generate anti-PRRSV pigs.In this study,we first generated immortalized porcine alveolar macrophage(IPAM)cell lines harboring 40-residues(residues 523-562,including R561(arginine(R)at position 561))deletion of CD163.PRRSV infection experiments showed that these IPAM cell lines were completely resistant to PRRSV infection.We then generated cloned pigs carrying CD163-R561A(an arginine(R)to alanine(A)substitution at position 561 of CD163).PRRSV challenge experiments in porcine alveolar macrophages(PAMs)isolated from the CD163-R561A pigs showed significantly lower susceptibility to PRRSV than that of CD163-R561 PAMs.Through this study,we show that CD163523-562 contains essential residues for mediating PRRSV infection,and that CD163 R561 significantly contributes to PRRSV infection but is not essential for infection.These functional sites can therefore serve as new targets for understanding the mechanism of PRRSV infection.Furthermore,CD163-R561A pigs can be used as an important model for improving pig germplasm with resistance against PRRSV.

关键词： pigs porcine alveolar macrophages dual-sgRNA homology-directed repair PRRSV CD163

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The collagen type Ⅰ alpha 1 chain gene is an alternative safe harbor locus in the porcine genome

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Journal of Integrative Agriculture 2023年第1期22卷 202-213页

作者： XIANG Guang-ming ZHANG Xiu-ling xu chang-jiang FAN Zi-yao xu Kui WANG Nan WANG Yue CHE Jing-jing xu Song-song MU Yu-lian LI Kui LIU Zhi-guo State Key Laboratory of Animal Nutrition/Key Laboratory of Animal Genetics Breeding and Reproduction of Ministry of Agriculture and Rural Affairs of China Institute of Animal SciencesChinese Academy of Agricultural SciencesBeijing 100193P.R.China School of Life Science and Engineering Foshan UniversityFoshan 528231P.R.China Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs Agricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518120P.R.China

Efficient and stable expression of foreign genes in cells and transgenic animals is important for gain-of-function studies and the establishment of bioreactors.Safe harbor loci in the animal genome enable consistent overexpression of foreign genes,without side effects.However,relatively few safe harbor loci are available in pigs,a fact which has impeded the development of multi-transgenic pig research.We report a strategy for efficient transgene knock-in in the endogenous collagen type I alpha 1 chain(COL1A1)gene using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9)system.After the knock-in of a 2A peptide-green fluorescence protein(2A-GFP)transgene in the last codon of COL1A1 in multiple porcine cells,including porcine kidney epithelial(PK15),porcine embryonic fibroblast(PEF)and porcine intestinal epithelial(IPI-2I)cells,quantitative PCR(qPCR),Western blotting,RNA-seq and CCK8 assay were performed to assess the safety of COL1A1 locus.The qPCR results showed that the GFP knock-in had no effect(P=0.29,P=0.66 and P=0.20 for PK15,PEF and IPI-2I cells,respectively)on the mRNA expression of COL1A1 gene.Similarly,no significant differences(P=0.64,P=0.48 and P=0.80 for PK15,PEF and IPI-2I cells,respectively)were found between the GFP knock-in and wild type cells by Western blotting.RNA-seq results revealed that the transcriptome of GFP knock-in PEF cells had a significant positive correlation(P<2.2e–16)with that of the wild type cells,indicating that the GFP knock-in did not alter the global expression of endogenous genes.Furthermore,the CCK8 assay showed that the GFP knock-in events had no adverse effects(P_(24)h=0.31,P_(48)h=0.96,P_(72)h=0.24,P_(96)h=0.17,and P_(120)h=0.38)on cell proliferation of PK15 cells.These results indicate that the COL1A1 locus can be used as a safe harbor for foreign genes knock-in into the pig genome and can be broadly applied to farm animal breeding and biomedical model establishment.

关键词： COL1A1 gene safe harbor knock-in CRISPR/Cas9 pig

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Nonlinear Memoryless Channel Modeling Based on Wavelet

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The Journal of China Universities of Posts and Telecommunications 1999年第2期6卷 14-16,21页

作者： xu chang-jiang FENG Guang-zeng(Department of Telecommunication Engineering, Nanjing University of Posts and Telecommunications. Naming. 210003. P. R, China) Department of Telecommunication Engineering Nanjing University of Posts and Telecommunications Nanjing 210003 P. R. China

This paper presents a wavelet network for modeling nonlinear memoryless channels. In particular, thepaper deals with the approximation of the nonlinear characteristics of traveling-wave tube (TWT) amplifiers used insatellite communications. Simulation results demonstrate that wavelet network models can track adaptively a changeIn the TWT characteristics.

关键词： wavelet networks TWT amplifiers satellite communications

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Controllable chemical redox reactions to couple microbial degradation for organic contaminated sites remediation:A review

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Journal of Environmental Sciences 2024年第5期139卷 428-445页

作者： Bin Chen jiang xu Lizhong Zhu College of Environmental and Resource Sciences Zhejiang UniversityHangzhou 310058China Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control Zhejiang UniversityHangzhou 310058China Zhejiang Agriculture&Forest University Lin'an 311300China

Global environmental concern over organic contaminated sites has been progressively conspicuous during the process of urbanization and industrial restructuring.While traditional physical or chemical remediation technologies may significantly destroy the soil structure and function,coupling moderate chemical degradation with microbial remediation becomes a potential way for the green,economic,and efficient remediation of contaminated sites.Hence,this work systematically elucidates why and how to couple chemical technology with microbial remediation,mainly focused on the controllable redox reactions of organic contaminants.The rational design of materials structure,selective generation of reactive oxygen species,and estimation of degradation pathway are described for chemical oxidation.Meanwhile,current progress on efficient and selective reductions of organic contaminants(i.e.,dechlorination,defluorination,-NO_(2) reduction)is introduced.Combined with the microbial remediation of contaminated sites,several consideration factors of how to couple chemical and microbial remediation are proposed based on both fundamental and practical points of view.This review will advance the understanding and development of chemical-microbial coupled remediation for organic contaminated sites.

关键词： Soil remediation Groundwater remediation Rational design Reactive species Environmental nanotechnology

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METTL1 promotes colorectal cancer cell proliferation by attenuating CHEK2-induced G1/S phase arrest

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Genes & Diseases 2024年第2期11卷 579-581页

作者： Houxiang jiang Ying Liu Huibin Song Jinquan Xia Ying Tian Luolin Wang Mingwei Li Zhenyu xu Zhenglei xu chang Zou Department of Gastrointestinal Surgery The First Affiliated Hospital of Wannan Medical College (Yijishan Hospital of Wannan Medical College)WuhuAnhui 241001China Anhui Province Clinical Research Center for Critical Respiratory Medicine WuhuAnhui 241001China Department of Clinical Medical Research Center The Second Clinical Medical College (Shenzhen People's Hospital) of Jinan UniversityThe First Affiliated Hospital of Southern University of Science and TechnologyShenzhenGuangdong 518020China Integrated Chinese and Western Medicine Postdoctoral Research Station Jinan UniversityGuangzhouGuangdong 510632China School of Pharmacy Southwest Medical UniversityLuzhouSichuan 646000China Department of Gastroenterology Shenzhen People's Hospital(The Second Clinical Medical CollegeJinan UniversityThe First Affiliated HospitalSouthern University of Science and Technology)ShenzhenGuangdong 518020China Precision Medicine Center The First Affiliated Hospital of Wannan Medical College(Yijishan Hospital of Wannan Medical College)WuhuAnhui 241001China School of Medicine Life and Health SciencesThe Chinese University of Hong KongShenzhenGuangdong 518172China

Colorectal carcinoma(CRC),the third most commonly diagnosed cancer,accounts for 9.7%of all newly diagnosed cancer cases and 9.4%of cancer-related deaths globally.1 Recent studies have demonstrated that post-transcriptional RNA modifications,such as N^(6)-methyladenosine,N^(5)-methylcytosine,and N^(7)-methylguanosine,play critical roles in the regulation of mRNA stability and translation,primary microRNA processing,and lncRNA-protein complex that contributes to the progression of human cancer.

关键词： cancer colorectal globally

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Association between Serum Ferritin Levels and Metabolic-associated Fatty Liver Disease in Adults:a Cross-sectional Study Based on the NHANES

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Current Medical Science 2024年第3期44卷 494-502页

作者： jiang-hui LI xue-yao MA Yun YI Lu-rao LI Zhi-yong xu Ying chang Department of Gastroenterology Zhongnan Hospital of Wuhan UniversityWuhan430071China Hubei Clinical Center and Key Laboratory of Intestinal and Colorectal Diseases Wuhan430071China Endoscopy Center The People’s Hospital of YingshanHuanggang438799China

Objective Ferritin,initially acting as an iron-storage protein,was found to be associated with metabolic diseases.Our study was designed to investigate the association between serum ferritin and metabolic-associated fatty liver disease(MAFLD)using data from the National Health and Nutrition Examination Survey(NHANES)of the United State of America.Methods A cross-sectional study was conducted,enrolling a total of 2145 participants from the NHANES in the 2017–2018 cycles.Hepatic steatosis and liver fibrosis were assessed by ultrasound images and several non-invasive indexes.Multiple regression analysis was conducted to determine the associations between serum ferritin concentration and MAFLD and liver fibrosis.Results The analysis revealed that participants with higher serum ferritin levels(Q3 and Q4 groups)had a higher prevalence of MAFLD than those with the lowest serum ferritin levels[Q3 vs.Q1:OR=2.17(1.33,3.53),P<0.05 in fatty liver index(FLI);Q4 vs.Q1:OR=3.13(1.91,5.13),P<0.05 in FLI].Additionally,participants with the highest serum ferritin levels(Q4 group)displayed a higher prevalence of liver fibrosis[Q4 vs.Q1:OR=2.59(1.19,5.62),P<0.05 in liver stiffness measurement;OR=5.06(1.12,22.94),P<0.05 in fibrosis-4 index],with significantly increased risk observed in participants with concomitant diabetes[OR=7.45(1.55,35.72),P=0.012].Conclusion Our study revealed that elevated serum ferritin levels are associated with a higher prevalence of MAFLD and advanced liver fibrosis in patients.Elevated serum ferritin levels combined with diabetes are important risk factors for liver fibrosis.

关键词： serum ferritin liver fibrosis metabolic-associated fatty liver disease National Health and Nutrition Examination Survey

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Developmental Patterns and Gender Differences of Vocal Production in Marmoset Monkeys

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Neuroscience Bulletin 2024年第1期40卷 133-138页

作者： Ruixin An Chaocheng Lu Chen Wang Liangtang chang Junfeng Huang Fan jiang Tian-Le xu Neng Gong Center for Brain Science Shanghai Children's Medical CenterShanghai Jiao Tong University School of MedicineShanghai 200127China Department of Anatomy and Physiology Shanghai Jiao Tong University School of MedicineShanghai 200025China Department of Critical Care Medicine Huashan HospitalFudan University12 Middle Wulumuqi RoadShanghai 200040China Institute of Neuroscience Key Laboratory of Primate NeurobiologyCAS Center for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesShanghai 200031China Lingang Laboratory Shanghai 200031China Shanghai Center for Brain Science and Brain-Inspired Technology Shanghai 201210China

Dear Editor,The common marmoset(Callithrix jacchus),a small New World monkey species,is considered an ideal non-human primate model for studying the evolution and neural mechanism of vocal behavior due to their rich call repertoire and vocal communication[1-3].Especially,the early vocal development of marmoset monkeys has attracted much attention,because newborn marmosets have been shown to exhibit a vocal development trajectory.Within two months after birth,marmosets'vocalizations undergo dramatic changes from cries to adult-like mature calls,a process that is influenced not only by inner physical maturation but also by parental care and feedback[4].

关键词： parental birth Pattern

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Ionic Liquid-Enhanced Assembly of Nanomaterials for Highly Stable Flexible Transparent Electrodes

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Nano-Micro Letters 2024年第7期16卷 441-455页

作者： Jianmin Yang Li chang Xiqi Zhang Ziquan Cao Lei jiang Key Laboratory of Bio-Inspired Materials and Interfacial Science Technical Institute of Physics and ChemistryChinese Academy of SciencesBeijing 100190People’s Republic of China University of Chinese Academy of Sciences Beijing 100049People’s Republic of China College of Chemistry and Chemical Engineering Chongqing University of TechnologyChongqing 400054People’s Republic of China Binzhou Institute of Technology Binzhou 256600People’s Republic of China Nanomics Biotechnology Co. Ltd.HangzhouPeople’s Republic of China

The controlled assembly of nanomaterials has demon-strated significant potential in advancing technological devices.However,achieving highly efficient and low-loss assembly technique for nanomate-rials,enabling the creation of hierarchical structures with distinctive func-tionalities,remains a formidable challenge.Here,we present a method for nanomaterial assembly enhanced by ionic liquids,which enables the fabrication of highly stable,flexible,and transparent electrodes featuring an organized layered structure.The utilization of hydrophobic and non-volatile ionic liquids facilitates the production of stable interfaces with water,effectively preventing the sedimentation of 1D/2D nanomaterials assembled at the interface.Furthermore,the interfacially assembled nanomaterial monolayer exhibits an alternate self-climbing behavior,enabling layer-by-layer transfer and the formation of a well-ordered MXene-wrapped silver nanowire network film.The resulting composite film not only demonstrates exceptional photoelectric performance with a sheet resistance of 9.4Ωsq^(-1) and 93%transmittance,but also showcases remarkable environmental stability and mechanical flexibility.Particularly noteworthy is its application in transparent electromagnetic interference shielding materials and triboelectric nanogenerator devices.This research introduces an innovative approach to manufacture and tailor functional devices based on ordered nanomaterials.

关键词： Ionic liquids Assembly Silver nanowires MXene nanosheets Flexible transparent electrodes

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Bromocriptine protects perilesional spinal cord neurons from lipotoxicity after spinal cord injury

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Neural Regeneration Research 2024年第5期19卷 1142-1149页

作者： Ying Peng Zhuoxuan Li Zhiyang Zhang Yinglun Chen Renyuan Wang Nixi xu Yuanwu Cao chang jiang Zixian Chen Haodong Lin Trauma Center Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghaiChina Department of Orthopedics Zhongshan HospitalFudan UniversityShanghaiChina Department of Rehabilitation Medicine Shanghai Geriatric Medical CenterShanghaiChina

Recent studies have revealed that lipid droplets accumulate in neurons after brain injury and evoke lipotoxicity,damaging the neurons.However,how lipids are metabolized by spinal cord neurons after spinal cord injury remains unclear.Herein,we investigated lipid metabolism by spinal cord neurons after spinal cord injury and identified lipid-lowering compounds to treat spinal cord injury.We found that lipid droplets accumulated in perilesional spinal cord neurons after spinal cord injury in mice.Lipid droplet accumulation could be induced by myelin debris in HT22 cells.Myelin debris degradation by phospholipase led to massive free fatty acid production,which increased lipid droplet synthesis,β-oxidation,and oxidative phosphorylation.Excessive oxidative phosphorylation increased reactive oxygen species generation,which led to increased lipid peroxidation and HT22 cell apoptosis.Bromocriptine was identified as a lipid-lowering compound that inhibited phosphorylation of cytosolic phospholipase A2 by reducing the phosphorylation of extracellular signal-regulated kinases 1/2 in the mitogen-activated protein kinase pathway,thereby inhibiting myelin debris degradation by cytosolic phospholipase A2 and alleviating lipid droplet accumulation in myelin debris-treated HT22 cells.Motor function,lipid droplet accumulation in spinal cord neurons and neuronal survival were all improved in bromocriptine-treated mice after spinal cord injury.The results suggest that bromocriptine can protect neurons from lipotoxic damage after spinal cord injury via the extracellular signal-regulated kinases 1/2-cytosolic phospholipase A2 pathway.

关键词： bromocriptine central nervous system cytosolic phospholipase A2 high-content screening lipid droplet lipid metabolism lipotoxicity mitogen-activated protein kinase spinal cord injury spinal cord neurons

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Gut microbial interactions based on network construction and bacterial pairwise cultivation

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Science China Life Sciences 2024年第8期 1751-1762页

作者： Min-Zhi jiang chang Liu chang xu He jiang Yulin Wang Shuang-jiang Liu State Key Laboratory of Microbial Technology Shandong University State Key Laboratory of Microbial Resources and Environmental Microbiology Research Center (EMRC)Institute of MicrobiologyChinese Academy of Sciences University of Chinese Academy of Sciences

Association networks are widely applied for the prediction of bacterial interactions in studies of human gut microbiomes.However,the experimental validation of the predicted interactions is challenging due to the complexity of gut microbiomes and the limited number of cultivated bacteria.In this study,we addressed this challenge by integrating in vitro time series network(TSN) associations and cocultivation of TSN taxon pairs.Fecal samples were collected and used for cultivation and enrichment of gut microbiome on YCFA agar plates for 13 days.Enriched cells were harvested for DNA extraction and metagenomic sequencing.A total of 198 metagenome-assembled genomes(MAGs) were recovered.Temporal dynamics of bacteria growing on the YCFA agar were used to infer microbial association networks.To experimentally validate the interactions of taxon pairs in networks,we selected 24 and 19 bacterial strains from this study and from the previously established human gut microbial biobank,respectively,for pairwise co-cultures.The co-culture experiments revealed that most of the interactions between taxa in networks were identified as neutralism(51.67%),followed by commensalism(21.67%),amensalism(18.33%),competition(5%) and exploitation(3.33%).Genome-centric analysis further revealed that the commensal gut bacteria(helpers and beneficiaries) might interact with each other via the exchanges of amino acids with high biosynthetic costs,short-chain fatty acids,and/or vitamins.We also validated 12 beneficiaries by adding 16 additives into the basic YCFA medium and found that the growth of 66.7% of these strains was significantly promoted.This approach provides new insights into the gut microbiome complexity and microbial interactions in association networks.Our work highlights that the positive relationships in gut microbial communities tend to be overestimated,and that amino acids,short-chain fatty acids,and vitamins are contributed to the positive relationships.

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