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Activatable chemiluminescence probe based on four-arm PEGconjugated-pyropheophorbide-a for in vivo autofluorescence-free imaging of peroxynitrite

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Science China Chemistry 2025年

作者： Jianmei Ke Lei Ding Ronggui Lian Cheng Zheng Wenjun Li Aixian Zheng Yupeng Sun Ming Wu Yongyi Zeng Xiaolong Liu Maochun Hong Xiaolong Zhang The United Innovation of Mengchao Hepatobiliary Technology key laboratory of Fujian Province Mengchao Hepatobiliary Hospital of Fujian Medical University Ganjiang Innovation Academy Chinese Academy of Sciences School of Rare Earths University of Science and Technology of China College of Chemistry Fuzhou University State key laboratory of Structure Chemistry Fujian Institute of Research on the Structure of Matter Chinese Academy of Sciences

Peroxynitrite（ONOO-） is a highly reactive nitrogen species that plays pivotal roles in cell signal transduction and physiological or pathological progresses. However, commonly used ONOO-optical imaging probes are still hampered by high background/autofluorescence（fluorescence probe）, short emission wavelength, or poor selectivity in the case of chemiluminescence. Herein,we report a facile method to prepare an activatable chemiluminescence probe（PPA-PEG） with good biocompatibility and functionality for in vivo autofluorescence-free imaging of ONOO-. The PPA-PEG consists of pyropheophorbide-a（PPA）, a typical deep red photosensitizer that acts as both the recognition and signaling element, and 4-arm poly（ethylene glycol）(4-arm PEG), which improves the biosafety and water solubility of probe. These components can self-assemble into nanoparticles（namely PPA-PEG nanoprobe） in aqueous solution. The PPA-PEG nanoprobe showed an ultra-low chemiluminescence signal before interacting with ONOO-, but exhibited good selectivity, high sensitivity and a fast response toward ONOO-. The PPAPEG was successfully applied to image cellular ONOO-changes, as well as the endogenous ONOO-changes in inflammation models and subcutaneous or orthotopic hepatocellular carcinoma（HCC） tumors models in living mice. In vitro and in vivo studies verified the good detection and imaging capabilities of PPA-PEG for peroxynitrite, demonstrating suitable tissue penetration and a high signal-to-background ratio（SBR）. Thus, our nanoprobe can serve as a valuable activatable chemiluminescence imaging tool for studying important peroxynitrite-related chemical and biological applications.

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Contribution of fungal biomass to persistent soil carbon across natural ecosystems

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Science China Earth Sciences 2025年

作者： Xiang WANG Guang-Hui YU Yakov KUZYAKOV Bo-Hao YIN Andreas KAPPLER Cong-Qiang LIU Bohai Coastal Critical Zone National Observation and Research Station Institute of Surface-Earth System Science School of Earth System Science Tianjin Key Laboratory of Earth Critical Zone Science and Sustainable Development in Bohai Rim Tianjin University Department of Soil Science of Temperate Ecosystems Department of Agricultural Soil Science University of G?ttingen Agro-Technological Institute Peoples Friendship University of Russia (RUDN University) Geomicrobiology Center for Applied Geosciences University of Tübingen Cluster of Excellence EXC 2124 Controlling Microbes to Fight Infection University of Tübingen

Fungi are known to be crucial in the formation and stabilization of soil organic matter through their exudates and dead residues (necromass). Yet, it remains unclear how fungi contribute to the persistence of carbon in soils over millennial scales across biomes. Here, this study fills the knowledge gap by linking fungal carbon stocks with minerals-associated carbon stocks across major biomes. A strong correlation between fungal biomass and reactive mineral-associated carbon stocks in soils across six biomes supports the substantial role of fungi in soil carbon persistence at the global level. High spatial resolution nanoscale secondary ion mass spectrometry revealed that fungi may stabilize carbon by forming organo-mineral associations through their close physical connection to reactive minerals and weathered nanoparticles. We propose a conceptual model that emphasizes the dualistic role of hypha-mineral interactions:(i) fungi accelerate organic matter decomposition by producing reactive oxygen species on the hypha-mineral interfaces, and (ii) fungi stabilize their residues on mineral surfaces, leading to millennial scale persistence of soil C. Concluding, fungi play a fundamental role in ecosystems that extends beyond the decomposition of persistent carbon. They are crucial in stabilizing carbon on mineral surfaces, thereby facilitating the long-term removal of carbon from rapid biotic cycling.

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SRP54 Negatively Regulates IFN-Beta Production and Antiviral Response by Targeting RIG-I and MDA5

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Virologica Sinica 2021年第2期36卷 231-240页

作者： Dong-Peng Wang Hong-Yan Zhang Bo-Wei Liao Zhen Tong Zhi-Sheng Xu Yan-Yi Wang Yan Yang key laboratory of Special Pathogens and biosafety Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan 430071China University of Chinese Academy of Sciences Beijing 100049China

During virus infection,RIG-I-like receptors(RLRs)recognize viral RNAs and recruit the adaptor protein VISA to activate downstream signaling,leading to activation of transcription factors NF-κB and IRF3,which collaborate to induce type I interferons(IFNs).IFNs further induce expression of hundreds of IFN-stimulated genes(ISGs)that suppress viral replication and facilitate the adaptive immune *** production of IFNs is implicated in various immune *** we identified Signal Recognition Particle 54(SRP54)as a negative regulator of RLRs-induced antiviral *** of SRP54 inhibited RNA virus-triggered induction of IFN-b and increased viral replication,whereas knockdown of SRP54 had opposite ***,SRP54 interacted with both RIG-I and MDA5 and impaired their association with *** findings demonstrate that SRP54 acts as a negative regulator of RLRs-mediated innate immune response by disrupting the recruitment of VISA to RIG-I/MDA5.

关键词： TypeⅠinterferons(IFNs) SRP54 RLRs-induced signaling RIG-I/MDA5 and VISA interactions

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Development and characterization of a clinical strain of Coxsackievirus A16 and an eGFP infectious clone

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Virologica Sinica 2015年第4期30卷 269-276页

作者： Chenglin Deng Xiaodan Li Siqing Liu Linlin Xu Hanqing Ye Cheng-Feng Qin Bo Zhang key laboratory of Special Pathogens and biosafety Wuhan Institute of Virology Chinese Academy of Sciences University of Chinese Academy of Sciences State key laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology

Coxsackievirus A16(CA16) is one of the major causes of hand, foot, and mouth disease(HFMD) worldwide, which is a common illness that affects children. The frequent occurrence of HFMD outbreaks has become a serious public health problem in Asia. Therefore, it is important to understand the pathogenesis and replication of CA16. In this study, a stable infectious c DNA clone of an epidemic strain of Coxsackievirus A16(CA16) was assembled, and subsequently a reporter virus(e GFP-CA16) was constructed by inserting the e GFP gene between the 5'-UTR and the N-terminus of VP4, with the addition of a 2A protease cleavage site(ITTLG) at its C-terminus. This was transfected into Vero cells to generate infectious recombinant viruses. The growth characteristics and plaque morphology, in vitro, in mammalian cells were found to be indistinguishable between the parental and recombinant viruses. Although the e GFP-CA16 showed smaller plaque size as compared to recombinant CA16, both were found to exhibit similar growth trends and EC50 of NITD008. In summary, this stable infectious c DNA clone should provide a valuable experimental system to study CA16 infection and host response. The e GFP-CA16 is expected to provide a powerful tool to monitor e GFP expression in infected cells and to evaluate the antiviral activity of potential antiviral agents in the treatment of CA16 infections.

关键词： Coxsackievirus A16(CA16) infectious cDNA clone rep

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Characteristics of SARS-CoV-2 transmission in a medium-sized city with traditional communities during the early COVID-19 epidemic in China

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Virologica Sinica 2022年第2期37卷 187-197页

作者： Yang Li Hao-Rui Si Yan Zhu Nan Xie Bei Li Xiang-Ping Zhang Jun-Feng Han Hong-Hong Bao Yong Yang Kai Zhao Zi-Yuan Hou Si-Jia Cheng Shuan-Hu Zhang Zheng-Li Shi Peng Zhou CAS key laboratory of Special Pathogens and biosafety Wuhan Institute of VirologyChinese Academy of SciencesWuhan430071China Anyang municipal Center for Disease Control and Prevention Anyang455000China University of Chinese Academy of Sciences Beijing101409China

The nationwide COVID-19 epidemic ended in 2020, a few months after its outbreak in Wuhan, China at the end of2019. Most COVID-19 cases occurred in Hubei Province, with a few local outbreaks in other provinces of China. A few studies have reported the early SARS-Co V-2 epidemics in several large cities or provinces of China. However,information regarding the early epidemics in small and medium-sized cities, where there are still traditionally large families and community culture is more strongly maintained and thus, transmission profiles may differ, is limited. In this study, we characterized 60 newly sequenced SARS-Co V-2 genomes from Anyang as a representative of small and medium-sized Chinese cities, compared them with more than 400 reference genomes from the early outbreak, and studied the SARS-Co V-2 transmission profiles. Genomic epidemiology revealed multiple SARS-Co V-2 introductions in Anyang and a large-scale expansion of the epidemic because of the large family ***, our study revealed two transmission patterns in a single outbreak, which were attributed to different social activities. We observed the complete dynamic process of single-nucleotide polymorphism development during community transmission and found that intrahost variant analysis was an effective approach to studying cluster infections. In summary, our study provided new SARS-Co V-2 transmission profiles representative of small and medium-sized Chinese cities as well as information on the evolution of SARS-Co V-2 strains during the early COVID-19 epidemic in China.

关键词： SARS-CoV-2 Epidemiology Community transmission Single-nucleotide polymorphism Intrahost variant

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Development of A MERS-CoV Replicon Cell Line for Antiviral Screening

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Virologica Sinica 2021年第4期36卷 730-735页

作者： Jing Chen Bing-Jie Hu Kai Zhao Yun Luo Hao-Feng Lin Zheng-Li Shi CAS key laboratory of Special Pathogens and biosafety Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China University of Chinese Academy of Sciences Beijing100049China

Middle East respiratory syndrome coronavirus(MERS-CoV)is the causative agent of a severe respiratory disease with a high mortality of~35%.The lack of approved treatments for MERS-CoV infection underscores the need for a user-friendly system for rapid drug *** this study,we constructed a MERS-CoV replicon containing the Renilla luciferase(Rluc)reporter gene and a stable luciferase replicon-carrying cell *** this cell line,we showed that MERS-CoV replication was inhibited by combined application of lopinavir and ritonavir,indicating that this cell line can be used to screen inhibitors of MERS-CoV ***,the MERS-replicon cell line can be used for high-throughput screening of antiviral drugs without the need for live virus handling,providing an effective and safe tool for the discovery of antiviral drugs against MERS-CoV.

关键词： Middle East respiratory syndrome coronavirus(MERS-CoV) Replicon cell line Antiviral screening Luciferase reporter gene

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Identification and Characterization of a Novel Single Domain Antibody Against Ebola Virus

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Virologica Sinica 2021年第6期36卷 1600-1610页

作者： Rui Wang Haiwei Zhang Cheng Peng Jian Shi Huajun Zhang Rui Gong CAS key laboratory of Special Pathogens and biosafety Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China University of Chinese Academy of Sciences Beijing100049China

Ebola virus(EBOV) belongs to the Filoviridae family and causes severe illnesses such as hemorrhagic fever with a high mortality rate up to 90%. Now two antibody drugs termed Inmazeb and Ebanga have been approved for treating EBOV infection. However, clinical studies have demonstrated that the mortality rate of the patients who received these two antibody drugs remains above 30%. Therefore, novel therapeutics with better efficacy is still desired. The isolated human IgG1 constant domain 2(CH2 domain) has been proposed as a scaffold for the development of C-based single domain antibodies(C-sd Abs) as therapeutic candidates against viral infections and other diseases. Here, we screened and identified a novel C-sd Ab termed M24 that targets EBOV glycoprotein(GP) from a C-sd Ab phage display library. M24 neutralizes the pseudotype EBOV with IC;of 0.8 nmol/L(12 ng/mL) and has modest neutralizing activity against authentic *** determination, including molecular docking and site mutation analysis, discloses that M24 binds to the internal fusion loop(IFL) within GP2, a transmembrane subunit of GP. Interestingly, we found that the binding of M24 to GP at pH5.5 has dramatically decreased compared to the binding at pH 7.5, which may lead to weak efficacy in the neutralization of authentic EBOV. Since no sd Ab against EBOV infection has been reported to date, our results not only give a proof of concept that sd Abs could be utilized for the development of potential therapeutic candidates against EBOV infection, but also provide useful information for the discovery and improvement of anti-EBOV agents.

关键词： Ebola virus(EBOV) Constant domain 2(CH2 domain) C-based single domain antibody(C-sdAb) Neutralize Fusion loop

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Assessing risk of Bombali virus spillover to humans by mutagenesis analysis of viral glycoprotein

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hLife 2024年第1期2卷 32-42页

作者： Lincan Li Tianyu Gan Ziyue Ma Yi Huang Jin Zhong Unit of Viral Hepatitis CAS Key Laboratory of Molecular Virology and ImmunologyShanghai Institute of Immunity and InfectionCenter for Biosafety Mega-ScienceChinese Academy of SciencesShanghaiChina University of Chinese Academy of Sciences BeijingChina ShanghaiTech University ShanghaiChina Wuhan National biosafety laboratory Center for Biosafety Mega-ScienceChinese Academy of SciencesHubeiChina

Ebolaviruses,such as the highly pathogenic Ebola virus(EBOV),belong to the family Filoviridae and have caused out-breaks of severe and fatal hemorrhagic fever over the past *** virus(BOMV)is a newly discovered ebolavirus from bats with unknown potential to infect *** most ebolaviruses,the interaction between viral glycoprotein(GP)and host receptor Niemann-Pick C1(NPC1)is crucial for viral entry and determines host ***,we analyzed the BOMV GP-mediated virus entry into human cells using our recently developed EBOV transcription-and replication-competent virus-like particle(trVLP)*** demonstrated that while BOMV GP can be efﬁciently incorporated into trVLPs,it is inefﬁcient in mediating trVLP entry into human ***,BOMV GP-mediated virus entry into human cells can be signiﬁcantly enhanced by a few mutations in the NPC1-binding domain of ***,we showed that these mutations increase the binding of BOMV-GP to human *** summary,our results suggested that although wild-type BOMV does not efﬁciently infect human cells,the emergence of mutations in viral GP may boost its ability to spill over to humans,highlighting the importance of monitoring BOMV GP evolution in preventing potential BOMV spillover events.

关键词： Bombali virus glycoprotein entry Ebola virus Niemann-Pick C1

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Recovery of a Far-Eastern Strain of Tick-Borne Encephalitis Virus with a Full-Length Infectious cDNA Clone

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Virologica Sinica 2021年第6期36卷 1375-1386页

作者： Penghui Li Chen Yao Ting Wang Tong Wu Wenfu Yi Yue Zheng Yuanjiu Miao Jianhong Sun Zhongyuan Tan Yan Liu Xiaowei Zhang Hanzhong Wang Zhenhua Zheng CAS key laboratory of Special Pathogens and biosafety Wuhan Institute of VirologyCenter for Biosafety MegaScienceChinese Academy of SciencesWuhan 430071China College of Life Sciences and Food Engineering Hebei University of EngineeringHandan 056038China

Tick-borne encephalitis virus(TBEV) is a pathogenic virus known to cause central nervous system(CNS) diseases in humans, and has become an increasing public health threat nowadays. The rates of TBEV infection in the endemic countries are increasing. However, there is no effective antiviral against the disease. This underscores the urgent need for tools to study the emergence and pathogenesis of TBEV and to accelerate the development of vaccines and antivirals. In this study, we reported an infectious c DNA clone of TBEV that was isolated in China(the WH2012 strain). A beta-globin intron was inserted in the coding region of nonstructural protein 1(NS1) gene to improve the stability of viral genome in bacteria. In mammalian cells, the inserted intron was excised and spliced precisely, which did not lead to the generation of inserted mutants. High titers of infectious progeny viruses were generated after the transfection of the infectious clone. The cDNA-derived TBEV replicated efficiently, and caused typical cytopathic effect(CPE) and plaques in BHK-21 cells. In addition, the CPE and growth curve of cDNA-derived virus were similar to that of its parental isolate in cells. Together, we have constructed the first infectious TBEV cDNA clone in China, and the clone can be used to investigate the genetic determinants of TBEV virulence and disease pathogenesis, and to develop countermeasures against the virus.

关键词： Tick-borne encephalitis virus(TBEV) Infectious cDNA clone Intron Virus replication

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Isolation and Characterization of Monkeypox Virus from the First Case of Monkeypox—Chongqing municipality,China,2022

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China CDC weekly 2022年第46期4卷 1019-1024页

作者： Baoying Huang Hua Zhao Jingdong Song Li Zhao Yao Deng Wen Wang Roujian Lu Wenling Wang Jiao Ren Fei Ye Houwen Tian Guizhen Wu Hua Ling Wenjie Tan key laboratory of biosafety National Health and Family Planning CommissionNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing MunicipalityChina Chongqing Center for Disease Control and Prevention Chongqing Municipal Key Laboratory for High Pathogenic MicrobesChongqing MunicipalityChina

Introduction:The first imported case of monkeypox(MPX)from the mainland of China was reported in September ***,the study reports the isolation and characterization of MPX virus(MPXV)in this ***:Clinical specimens including skin blister fluid,oropharyngeal and nasopharyngeal swabs,and blood were collected and inoculated onto Vero *** isolated virus was identified as MPXV using quantitative polymerase chain reaction(qPCR),cytopathic effects(CPEs),immunofluorescence assay(IFA)and transmission electron microscopy(TEM).Plaque assays were employed to quantify infectious plaque-forming units(PFUs).The plaque reduction neutralization test(PRNT)was developed to determine the neutralizing antibody(nAb)against ***:MPXV replication was confirmed with *** CPEs were observed 48 h *** isolated virus was named *** showed that MPXV reacted with serum of MPX *** morphology was observed using *** virus titer increased to>106 PFU/mL after three *** serum PRNT 50%neutralization titer(NT50)was 35 for the MPX patient 6 days after symptom ***:The study successfully isolated the first MPXV strain in the mainland of China,*** titration and PRNT methods have been *** study provides key resources and technical platforms for further research as well as anti-viral drug and vaccine development against MPX.

关键词： mainland Chongqing fir

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