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A new anterograde trans-synaptic tracer based on Sindbis virus

Neural Regeneration Research 2022年第12期17卷 2761-2764页

作者： Xiang-Wei Shi Fan Jia Pei Lyu fu-qiang xu State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics Key Laboratory of Magnetic Resonance in Biological SystemsWuhan Center for Magnetic ResonanceInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of SciencesWuhanHubei ProvinceChina Guangdong Provincial Key Laboratory of Brain Connectome and Behavior CAS Key Laboratory of Brain Connectome and Manipulationthe Brain Cognition and Brain Disease Institute(BCBDI)Translational Research Center for the Nervous System(TRCNS)Shenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhenGuangdong ProvinceChina NMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal Products Key Laboratory of Quality Control Technology for Virus-Based TherapeuticsGuangdong Provincial Medical Products AdministrationShenzhen Key Laboratory of Viral Vectors for BiomedicineShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhenGuangdong ProvinceChina University of Chinese Academy of Sciences BeijingChina Wuhan National Laboratory for Optoelectronics Huazhong University of Science and TechnologyWuhanHubei ProvinceChina

Mapping neural circuits is critical for understanding the structure and function of the nervous system.Engineered viruses are a valuable tool for tracing neural circuits.However,current tracers do not fully meet the needs for this approach because of various drawbacks,such as toxicity and characteristics that are difficult to modify.Therefore,there is an urgent need to develop a new tracer with low toxicity and that allows for long-term studies.In this study,we constructed an engineered Sindbis virus(SINV)expressing enhanced green fluorescent protein(EGFP)reporter gene(SINV-EGFP)and found that it had no significant difference in biological characterization compared with the wild-type Sindbis virus in BHK-21 cells and neurons in vitro.We injected the virus into the visual circuit of mouse brain and found that the virus infected neurons in the local injected site and anterogradely spread in the neural circuits.Although the efficiency of transmission was limited,the findings demonstrate that SINV can be used as a new anterograde tracer to map neural circuits in mouse brain and that it spreads exclusively in the anterograde direction.further,use of SINV in mouse brain research will provide longer time windows for circuit tracing than is possible with herpes simplex virus and vesicular stomatitis virus tracers.

关键词： anterograde lateral geniculate nucleus mouse brains neural circuit neurons retina Sindbis virus superior colliculus synapse tracer

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Anatomical Evidence for the Neural Connection from the Emotional Brain to Autonomic Innervation in the Anterior Chamber Structures of the Eye

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Current Medical Science 2022年第2期42卷 417-425页

作者： Lin MA Fang YANG Qing LIU xu-tao ZHU Xin LIU Sen JIN Hua-dong WANG Lei PEI fu-qiang xu Hai-xia LIU Department of Ophthalmology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430033China Shenzhen Key Laboratory of Viral Vectors for Biomedicine the Brain Cognition and Brain Disease Institute(BCBDI)Shenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhen 518055China University of Chinese Academy of Sciences Beijing100049China State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics Key Laboratory of Magnetic Resonance in Biological SystemsWuhan Center for Magnetic ResonanceInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of SciencesWuhan430071China The Institute for Brain Research Collaborative Innovation Center for Brain ScienceHuazhong University of Science and TechnologyWuhan430074China

Objective Previous studies have shown that the autonomic nervous system(ANS),which can be affected by emotions,is important in the occurrence or progression of glaucoma.The autonomic innervation distributed in the anterior chamber(AC)structures might play an efferent role in the neural regulation of intraocular pressure(IOP).This study aimed to investigate the anatomic neural connection from the emotional brain to autonomic innervation in the AC.Methods A retrograde trans-multisynaptic pseudorabies virus encoded with an enhanced green fluorescent protein(PRV531)and non-trans-synaptic tracer FAST Dil were injected into the right eye of mice,respectively.Fluorescent localization in the emotional brain and preganglionic nuclei was studied.Five and a half days after PRV531 injection into the right AC,fluorescent signals were observed in several emotional brain regions,including the amygdala,agranular insular cortex,lateral septal nuclei,periaqueductal gray,and hypothalamus.Autonomic preganglionic nuclei,including Edinger-Westphal nucleus,superior salivatory nucleus,and intermediolateral nucleus,were labeled using PRV531.Results The sensory trigeminal nuclei were not labeled using PRV531.The fluorescence signals in the nuclei mentioned above showed bilateral distribution,primarily on the ipsilateral side.Seven days after injecting FAST Dil into the AC,we observed no FAST Dil-labeled neurons in the central nervous system.Conclusion Our results indicate a neural connection from the emotional brain to autonomic innervation in the AC,which provides anatomical support for the emotional influence of IOP via the ANS.

关键词： intraocular pressure emotional brain amygdala anterior chamber autonomic innervation

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A rabies virus-based toolkit for efficient retrograde labeling and monosynaptic tracing

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Neural Regeneration Research 2023年第8期18卷 1827-1833页

作者： Kun-Zhang Lin Lei Li Wen-Yu Ma Xin Yang Zeng-Peng Han Neng-Song Luo Jie Wang fu-qiang xu The Brain Cognition and Brain Disease Institute(BCBDI) Shenzhen Key Laboratory of Viral Vectors for BiomedicineShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsNMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal ProductsShenzhenKey Laboratory of Quality Control Technology for Virus-Based TherapeuticsGuangdong Provincial Medical Products AdministrationShenzhenGuangdong ProvinceChina State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics National Center for Magnetic Resonance in WuhanWuhan Institute of Physics and MathematicsInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of Sciences-Wuhan National Laboratory for OptoelectronicsWuhanHubei ProvinceChina University of Chinese Academy of Sciences BeijingChina Wuhan National Laboratory for Optoelectronics Huazhong University of Science and TechnologyWuhanHubei ProvinceChina Shenzhen-Hong Kong Institute of Brain Science-Shenzhen fundamental Research Institutions ShenzhenGuangdong ProvinceChina Center for Excellence in Brain Science and Intelligence Technology Chinese Academy of SciencesShanghaiChina

Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.

关键词： functional studies neural activity neural circuits projection neurons rAAV9-Retro rabies virus recombination retrograde labeling synaptic inputs trans-monosynaptic tracing

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Co-localization of two-color rAAV2-retro confirms the dispersion characteristics of efferent projections of mitral cells in mouse accessory olfactory bulb

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Zoological Research 2020年第2期41卷 148-156页

作者： Ning Zheng Zhi-Zhong Wang Song-Wei Wang Fang-Jia Yang xu-Tao Zhu Chen Lu Anne Manyande Xiao-Ping Rao fu-qiang xu Center of Brain Science State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsNational Center for Magnetic Resonance in WuhanKey Laboratory of Magnetic Resonance in Biological SystemsWuhan Institute of Physics and MathematicsInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of SciencesWuhanHubei 430071China University of the Chinese Academy of Sciences Beijing 100049China Department of Automation School of Electrical EngineeringZhengzhou UniversityZhengzhouHenan 450001China School of Life Science Wuhan UniversityWuhanHubei 430072China School of Human and Social Sciences University of West LondonMiddlesex TW89GAUK Divisions of Biomedical Photonics Wuhan National Laboratory for OptoelectronicsWuhanHubei 430074China

The accessory olfactory bulb(AOB), located at the posterior dorsal aspect of the main olfactory bulb(MOB), is the first brain relay of the accessory olfactory system(AOS), which can parallelly detect and process volatile and nonvolatile social chemosignals and mediate different sexual and social behaviors with the main olfactory system(MOS). However, due to its anatomical location and absence of specific markers, there is a lack of research on the internal and external neural circuits of the AOB. This issue was addressed by singlecolor labeling and fluorescent double labeling using retrograde rAAVs injected into the bed nucleus of the stria terminalis(BST), anterior cortical amygdalar area(ACo), medial amygdaloid nucleus(MeA), and posteromedial cortical amygdaloid area(PMCo) in mice. We demonstrated the effectiveness of this AOB projection neuron labeling method and showed that the mitral cells of the AOB exhibited efferent projection dispersion characteristics similar to those of the MOB. Moreover, there were significant differences in the number of neurons projected to different brain regions, which indicated that each mitral cell in the AOB could project to a different number of neurons in different cortices. These results provide a circuitry basis to help understand the mechanism by which pheromone information is encoded and decoded in the AOS.

关键词： Accessory olfactory bulb Efferent projections Retrograde rAAVs Projection neuron labeling Dispersion characteristics Circuitry basis

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Reversal of Social Recognition Deficit in Adult Mice with MECP2 Duplication via Normalization of MeCP2 in the Medial Prefrontal Cortex

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Neuroscience Bulletin 2020年第6期36卷 570-584页

作者： Bin Yu Bo Yuan Jian-Kun Dai Tian-lin Cheng Sheng-Nan Xia Ling-Jie He Yi-Ting Yuan Yue-Fang Zhang Hua-Tai xu fu-qiang xu Zhi-Feng Liang Zi-Long Qiu Institute of Neuroscience State Key Laboratory of NeuroscienceChinese Academy of Sciences Center for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesShanghai 200031China University of Chinese Academy of Sciences Beijing 100049China Department of Molecular and Human Genetics Howard Hughes Medical InstituteBaylor College of MedicineHoustonTX 77030USA Wuhan Institute of Physics and Mathematics Chinese Academy of SciencesWuhan 430071China

Methyl-CpG binding protein 2(MeCP2) is a basic nuclear protein involved in the regulation of gene expression and microRNA processing.Duplication of MECP2-containing genomic segments causes MECP2 duplication syndrome,a severe neurodevelopmental disorder characterized by intellectual disability,motor dysfunction,heightened anxiety,epilepsy,autistic phenotypes,and early death.Reversal of the abnormal phenotypes in adult mice with MECP2 duplication(MECP2-TG) by normalizing the MeCP2 levels across the whole brain has been demonstrated.However,whether different brain areas or neural circuits contribute to different aspects of the behavioral deficits is still unknown.Here,we found that MECP2-TG mice showed a significant social recognition deficit,and were prone to display aversive-like behaviors,including heightened anxiety-like behaviors and a fear generalization phenotype.In addition,reduced locomotor activity was observed in MECP2-TG mice.However,appetitive behaviors and learning and memory were comparable in MECP2-TG and wild-type mice.functional magnetic resonance imaging illustrated that the differences between MECP2-TG and wild-type mice were mainly concentrated in brain areas regulating emotion and social behaviors.We used the CRISPR-Cas9 method to restore normal MeCP2 levels in the medial prefrontal cortex(mPFC) and bed nuclei of the stria terminalis(BST) of adult MECP2-TG mice,and found that normalization of MeCP2 levels in the mPFC but not in the BST reversed the social recognition deficit.These data indicate that the mPFC is responsible for the social recognition deficit in the transgenic mice,and provide new insight into potential therapies for MECP2 duplication syndrome.

关键词： MECP2 duplication Medial prefrontal cortex Social recognition deficit CRISPR-Cas9

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A high-throughput screening method for breeding Aspergillus niger with 12C6+ ion beam-improved cellulase

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Nuclear Science and Techniques 2017年第1期28卷 1-7页

作者： Bo-Ling Jiang Shu-Yang Wang Yu-Chen Wang Ji-Hong Chen Wen-Jian Li Jing Liu Wei Hu Guo-Qing Xiao Miao-Ying Dong fu-qiang xu Institute of Modern Physics Chinese Academy of Sciences509 Nanchang Rd Lanzhou 730000 China University of Chinese Academy of Sciences No. 19A Yuquan Road Beijing 100049 China Lanzhou University 222 South Tianshui RoadLanzhou 730000 Gansu Province China Gansu Agricultural University Lanzhou 730000 Gansu Province China

In this study,a high-throughput screening method was established through the 24-square deep-well microliter plate(MTP) fermentation and micro-plate detection for large-scale screening of the mutants.It was suitable for screening a large number of mutants and improving the breeding efficiency after heavy-ion beam irradiation.Seventeen strains showed higher cellulase activity compared with the initial strain after the screening of plate and MTP fermentation.The filter paper activity and β-glucosidase activity of Aspergillus niger H11201 had increased 38.74 and 63.23%separately compared with A.niger H11 by shaking flask fermentation,and it was genetically stable after being passaged to nine generations.The results indicate that the high-throughput screening method can be used for the quick breeding of A.niger with high cellulase activity.

关键词： High-throughput screening method Aspergillus niger H11 Cellulase 12C6+ ion beam

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Impact of initial fluctuations and nuclear deformations in isobar collisions

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Nuclear Science and Techniques 2024年第6期35卷 155-162页

作者： Jian-Fei Wang Hao-Jie xu fu-qiang Wang Department of Modern Physics University of Science and Technology of ChinaHefei 230026China School of Science Huzhou UniversityHuzhou 313000China Strong-Coupling Physics International Research Laboratory(SPiRL) Huzhou UniversityHuzhou 313000China Department of Physics and Astronomy Purdue UniversityWest LafayetteIN 47907USA

Relativistic isobar^(96)_(44)Ru+^(96)_(44)Ru and^(96)_(40)Zr+^(96)_(40)Zrcollisions have revealed intricate differences in their nuclear size and shape,inspiring unconventional studies of nuclear structure using relativistic heavy ion collisions.In this study,we investigate the relative differences in the mean multiplicityR_()and the secondR_(ε2)and third-order eccentricityR_(ε3)between isobar collisions using initial state Glauber models.It is found that initial fluctuations and nuclear deformations have negligible effects on R_()in most central collisions,while both are important for the R_(ε2)and R_(ε3),the degree of which is sensitive to the underlying nucleonic or sub-nucleonic degree of freedom.These features,compared to real data,may probe the particle production mechanism and the physics underlying nuclear structure.

关键词： Relativistic isobar collisions Initial fluctuations Nuclear deformation

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Ectomycorrhizal fungi and dark septate endophyte inoculation improve growth and tolerance of Pinus tabulaeformis under cadmium stress

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Pedosphere 2024年第2期34卷 473-483页

作者： Yong ZHOU Yanyan ZHENG Pengwei LI Lingjie xu qiang fu College of Landscape Architecture and Tourism Hebei Agricultural UniversityBaoding 071000China

Forest trees can establish symbiotic associations with dark septate endophytes(DSEs)and ectomycorrhizal fungi(ECMF)simultaneously.However,the combined effects of these two fungi on the growth and cadmium(Cd)tolerance of host plants remain largely unexplored.To address this knowledge gap,a pot experiment was conducted to examine the effects of the interaction between an ECMF strain(Suillus granulatus)and a DSE strain(Pseudopyrenochaeta sp.)on Pinus tabulaeformis under Cd stress,by assessing plant growth and physiological parameters,nutrient uptake,and soil properties.Notably,the colonization rates of both fungal strains were found to increase in response to Cd stress,with the extent of this increase being influenced by the specific fungal species and the Cd level in the soil.Compared to the non-inoculation treatment,single inoculation with fungal strain resulted in enhanced biomass,root development,and nutrient contents in P.tabulaeformis seedlings under Cd stress.furthermore,a synergistic effect was observed when these seedlings were co-inoculated with S.granulatus and Pseudopyrenochaeta sp.,as indicated by significantly greater measurements in various indicators compared to both the single and non-inoculation treatments.fungal inoculation effectively regulated the antioxidant defense responses and photosynthesis of P.tabulaeformis seedlings subjected to Cd stress,particularly in the co-inoculation treatment.In addition,fungal inoculation facilitated the Cd accumulation in P.tabulaeformis,suggesting a promising potential for the implementation of bioremediation strategies in the areas contaminated with heavy metals.The findings from this study indicate that the utilization of root symbiotic fungi obtained from stress environments could potentially enhance the growth performance and tolerance of P.tabulaeformis towards heavy metals,and co-inoculation of both fungal groups may result in even more pronounced synergistic effects on the overall fitness of the plant.

关键词： antioxidant defense response colonization rate fungal inoculation heavy metal tolerance physiological parameter root symbiotic fungi soil property

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Mucus hypersecretion in the airway

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Chinese Medical Journal 2008年第7期121卷 649-652页

作者： WANG Ke WEN fu-qiang xu Dan Division of Pulmonary Diseases State Key Laboratory of Biotherapy of China and Department of Respiratory MedicineWest China Hospital of Sichuan University Chengdu Sichuan610041 China

Mucus hypersecretion is a distinguishing feature of chronic inflammation diseases, such as asthma,chronic bronchitis, bronchiectasis and cystic fibrosis Mucus hypersecretion leads to impairment of mucociliary clearance, abnormal bacterial plantation, mucus plug in the airway, and dysfunction of gas exchange.5 To block this vicious cycle, chronic inflammation in the airway must be controlled and mucus hypersecretion must be reduced.

关键词： mucus hypersecretion airway

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Application of tissue engineering in the treatment of stress urinary incontinence

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Chinese Medical Journal 2009年第5期122卷 571-576页

作者： SONG Xiao-fei fu qiang xu Yue-min Department of Urology Shanghai Sixth People's Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200233 China

Stress urinary incontinence is one of the most common diseases in urology. The main treatments for stress urinary incontinence are pharmacotherapy, physicobehavioral therapy and surgery.However, the results of present methods are not satisfactory. Tissue engineering is a newly emerging technology that may provide a novel method for the treatment of stress urinary incontinence.

关键词： stress urinary incontinence tissue engineering muscle-derived stem cells adipose-derived stem cells myoblast fibroblast

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