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Development of a droplet digital polymerase chain reaction assay for the sensitive detection of total and integrated hiv-1 DNA

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Chinese Medical Journal 2024年第6期137卷 729-736页

作者： Lin Yuan Zhiying Liu Xin Zhang Feili Wei Shan Guo Na Guo Lifeng Liu Zhenglai Ma Yunxia Ji Rui Wang Xiaofan Lu Zhen Li Wei Xia Hao Wu Tong Zhang Bin Su Beijing Key Laboratory for hiv/aids Research Clinical and Research Center for Infectious DiseasesBeijing Youan HospitalCapital Medical UniversityBeijing 100069China Beijing institute of Hepatology Beijing Youan HospitalCapital Medical UniversityBeijing 100069China

Background:Total human immunodeficiency virus(hiv)DNA and integrated hiv DNA are widely used markers of hiv persistence.Droplet digital polymerase chain reaction(ddPCR)can be used for absolute quantification without needing a standard curve.Here,we developed duplex ddPCR assays to detect and quantify total hiv DNA and integrated hiv DNA.Methods:The limit of detection,dynamic ranges,sensitivity,and reproducibility were evaluated by plasmid constructs containing both the hiv long terminal repeat(LTR)and human CD3 gene(for total hiv DNA)and ACH-2 cells(for integrated hiv DNA).Forty-two cases on stable suppressive antiretroviral therapy(ART)were assayed in total hiv DNA and integrated hiv DNA.Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive(CD4^(+))T-cell counts,CD8^(+)T-cell counts and CD4/CD8 T-cell ratio,respectively.The assay linear dynamic range and lower limit of detection(LLOD)were also assessed.Results:The assay could detect the presence of hiv-1 copies 100%at concentrations of 6.3 copies/reaction,and the estimated LLOD of the ddPCR assay was 4.4 hiv DNA copies/reaction(95%confidence intervals[CI]:3.6-6.5 copies/reaction)with linearity over a 5-log_(10)-unit range in total hiv DNA assay.For the integrated hiv DNA assay,the LLOD was 8.0 copies/reaction(95%CI:5.8-16.6 copies/reaction)with linearity over a 3-log 10-unit range.Total hiv DNA in CD4^(+)T cells was positively associated with integrated hiv DNA(r=0.76,Phiv DNA and integrated hiv DNA in CD4^(+)T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8^(+)T-cell counts.Conclusions:This ddPCR assay can quantify total hiv DNA and integrated hiv DNA efficiently with robustness and sensitivity.It can be readily adapted for measuring hiv DNA with non-B clades,and it could be beneficial for testing in clinical trials.

关键词： Human immunodeficiency virus hiv Integrated hiv-1 DNA Total hiv DNA Droplet digital polymerase chain reaction hiv reservoir Antiretroviral therapy

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Overexpression of wild-type HRAS drives non-alcoholic steatohepatitis to hepatocellular carcinoma in mice

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Zoological Research 2024年第3期45卷 551-566页

作者： Chen Ling Su-Su Liu Yu-Ya Wang Gui-Tao Huo Yan-Wei Yang Nan Xu Hong Wang Yong Wu Yu-Fa Miao Rui Fu Yu-Wei Zhao Chang-Fa Fan College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceXi’anShaanxi 710069China Division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China National Center for Safety Evaluation of Drugs National Institutes for Food and Drug Control(NIFDC)Beijing 100176China Division of hiv/aids and Sexually Transmitted Virus Vaccines Institute for Biological Products ControlNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China Division of Laboratory Animal Monitoring Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)Beijing 102629China

Hepatocellular carcinoma(HCC),a prevalent solid carcinoma of significant concern,is an aggressive and often fatal disease with increasing global incidence rates and poor therapeutic outcomes.The etiology and pathological progression of non-alcoholic steatohepatitis(NASH)-related HCC is multifactorial and multistage.However,no single animal model can accurately mimic the full NASH-related HCC pathological progression,posing considerable challenges to transition and mechanistic studies.Herein,a novel conditional inducible wild-type human HRAS overexpressed mouse model(HRAS-HCC)was established,demonstrating 100%morbidity and mortality within approximately one month under normal dietary and lifestyle conditions.Advanced symptoms of HCC such as ascites,thrombus,internal hemorrhage,jaundice,and lung metastasis were successfully replicated in mice.In-depth pathological features of NASH-related HCC were demonstrated by pathological staining,biochemical analyses,and typical marker gene detections.Combined murine anti-PD-1 and sorafenib treatment effectively prolonged mouse survival,further confirming the accuracy and reliability of the model.Based on protein-protein interaction(PPI)network and RNA sequencing analyses,we speculated that overexpression of HRAS may initiate the THBS1-COL4A3 axis to induce NASH with severe fibrosis,with subsequent progression to HCC.Collectively,our study successfully duplicated natural sequential progression in a single murine model over a very short period,providing an accurate and reliable preclinical tool for therapeutic evaluations targeting the NASH to HCC continuum.

关键词： HRAS THBS1 HCC driver factor NASH Fibrosis Cirrhosis HCC Treatment evaluation

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LncRNA NKILA inhibits HBV replication by repressing NF-κB signalling activation

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Virologica Sinica 2024年第1期39卷 44-55页

作者： Xi Zhang Yuanyuan Li Chen Huan Yubao Hou Rujia Liu Hongyun Shi Peng Zhang Baisong Zheng Yingchao Wang Hong Wang Wenyan Zhang institute of Virology and aids Research The First Hospital of Jilin UniversityChangchun130012China Department of Ophthalmology The First Hospital of Jilin UniversityChangchun130012China Hepatobiliary Pancreatic Surgery The First Hospital of Jilin UniversityChangchun130012China

Hepatitis B virus(HBV)infection results in liver cirrhosis and hepatocellular carcinoma(HCC).HBx/nuclear factor(NF)-κB pathway plays a role in HBV replication.However,whether NF-κB-interacting long noncoding RNA(NKILA),a suppressor of NF-κB activation,regulates HBV replication remains largely unknown.In this study,gain-and-loss experiments showed that NKILA inhibited HBV replication by inhibiting NF-κB activity.In turn,HBV infection down-regulated NKILA expression.In addition,expression levels of NKILA were lower in the peripheral blood-derived monocytes(PBMCs)of HBV-positive patients than in healthy individuals,which were correlated with HBV viral loads.And a negative correlation between NKILA expression level and HBV viral loads was observed in blood serum from HBV-positive patients.Lower levels of endogenous NKILA were also observed in HepG2 cells expressing a 1.3-fold HBV genome,HBV-infected HepG2-NTCP cells,stable HBV-producing HepG2.2.15 and HepAD38 cells,compared to those HBV-negative cells.Furthermore,HBx was required for NKILA-mediated inhibition on HBV replication.NKILA decreased HBx-induced NF-κB activation by interrupting the interaction between HBx and p65,whereas NKILA mutants lack of essential domains for NF-κB inhibition,lost the ability to inhibit HBV replication.Together,our data demonstrate that NKILA may serve as a suppressor of HBV replication via NF-κB signalling.

关键词： LncRNA NF-κB-interacting longnoncoding RNA(NKILA) NF-ĸB signalling ChronichepatitisBvirus(HBV) Viral replication HBx

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AZD1775 and anti-PD-1 antibody synergistically sensitize hepatoma to radiotherapy

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Chinese Medical Journal 2024年第2期137卷 222-231页

作者： Yichun Yin Jian Wang Junxuan Yi Kaiyue Zhang Zimeng Yin Shunzi Jin Baisong Zheng Public Health of College Jilin UniversityJilinChangchun 130021China institute of Virology and aids Research The First Hospital of Jilin UniversityJilinChangchun 130021China NHC Key Laboratory of Radiobiology(Jilin University) JilinChangchun 130021China

Background:Radiation(IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment(TME).Wee1,a cell cycle regulator,can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1(CDK1).Meanwhile,programed death-1/programed death ligand-1(PD-1/PDL-1)blockade is closely related to TME.This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody(anti-PD-1 Ab)on radiosensitization of hepatoma.Methods:The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide(MTT)assay on human and mouse hepatoma cells HepG2,Hepa1-6,and H22.The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro.A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice,which were divided into control group,IR group,AZD1775 group,IR+AZD1775 group,IR+anti-PD-1 Ab group,and the IR+AZD1775+anti-PD-1 Ab group.Cytotoxic CD8^(+)T cells in TME were analyzed by flow cytometry.Results:Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro.AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage.AZD1775 treatment also reduced the proportion of PD-1^(+)/CD8^(+)T cells in the spleen of hepatoma subcutaneous xenograft mice.Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferonγ(IFNγ)^(+)or Ki67^(+)CD8 T cells and decreasing the levels of CD8^(+)Tregs cells in the tumor and spleen of the hepatoma mice model,indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγexpression,enhancing CD8^(+)T cells proliferation,and weakening CD8^(+)T cells depletion.Conclusions:This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improv

关键词： Hepatoma Radiation AZD1775 Anti-PD-1 antibody DNA damage Tumor microenvironment

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Uncovering the dominant contribution of intermediate volatility compounds in secondary organic aerosol formation from biomass-burning emissions

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National Science Review 2024年第3期11卷 150-158页

作者： Kun Li Jun Zhang David M.Bell Tiantian Wang Houssni Lamkaddam Tianqu Cui Lu Qi Mihnea Surdu Dongyu Wang Lin Du Imad El Haddad Jay G.Slowik Andre S.H.Prevot Environment Research institute Shandong University Laboratory of Atmospheric Chemistry Paul Scherrer Institute

Organic vapors from biomass burning are a major source of secondary organic aerosols(SOAs). Previous smog chamber studies found that the SOA contributors in biomass-burning emissions are mainly volatile organic compounds(VOCs). While intermediate volatility organic compounds(IVOCs) are efficient SOA precursors and contribute a considerable fraction of biomass-burning emissions, their contribution to SOA formation has not been directly observed. Here, by deploying a newly-developed oxidation flow reactor to study SOA formation from wood burning, we find that IVOCs can contribute ～70% of the formed SOA, i.e.>2 times more than VOCs. This previously missing SOA fraction is interpreted to be due to the high wall losses of semi-volatile oxidation products of IVOCs in smog chambers. The finding in this study reveals that SOA production from biomass burning is much higher than previously thought, and highlights the urgent need for more research on the IVOCs from biomass burning and potentially other emission sources.

关键词： atmospheric chemistry biomass burning secondary organic aerosol intermediate volatility organic compounds

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Motion of charged particles in bright squeezed vacuum

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Light(Science & Applications) 2024年第2期13卷 347-353页

作者： Matan Even Tzur Oren Cohen Solid State institute and Physics Department Technion-Israel Institute of TechnologyHaifa3200003Israel

The motion of laser-driven electrons quivers with an average energy termed pondermotive energy.We explore electron dynamics driven by bright squeezed vacuum(BSV),finding that BSV induces width oscillations,akin to electron quivering in laser light,with an equivalent ponderomotive energy.We identify closed and open trajectories of the electronic width that are associated with high harmonic generation and above-threshold ionization,respectively,similarly to trajectories of the electron position when its motion is driven by coherent light.In the case of bound electrons,the width oscillations may lead to ionization with noisy sub-cycle structure.Our results are foundational for strong-field and free-electron quantum optics,as they shed light on ionization,high harmonic generation,and nonlinear Compton scattering in BSV.

关键词： structure vacuum ionization

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On-Chip Micro Temperature Controllers Based on Freestanding Thermoelectric Nano Films for Low-Power Electronics

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Nano-Micro Letters 2024年第7期16卷 98-108页

作者： Qun Jin Tianxiao Guo Nicolas Perez Nianjun Yang Xin Jiang Kornelius Nielsch Heiko Reith institute for Metallic Materials Leibniz Institute for Solid State and Materials Research01069 DresdenGermany institute of Materials Engineering University of Siegen57076 SiegenGermany institute of Applied Physics Technical University of Dresden01069 DresdenGermany institute of Materials Science Technical University of Dresden01069 DresdenGermany

Multidimensional integration and multifunctional com-ponent assembly have been greatly explored in recent years to extend Moore’s Law of modern microelectronics.However,this inevitably exac-erbates the inhomogeneity of temperature distribution in microsystems,making precise temperature control for electronic components extremely challenging.Herein,we report an on-chip micro temperature controller including a pair of thermoelectric legs with a total area of 50×50μm^(2),which are fabricated from dense and flat freestanding Bi2Te3-based ther-moelectric nano films deposited on a newly developed nano graphene oxide membrane substrate.Its tunable equivalent thermal resistance is controlled by electrical currents to achieve energy-efficient temperature control for low-power electronics.A large cooling temperature difference of 44.5 K at 380 K is achieved with a power consumption of only 445μW,resulting in an ultrahigh temperature control capability over 100 K mW^(-1).Moreover,an ultra-fast cooling rate exceeding 2000 K s^(-1) and excellent reliability of up to 1 million cycles are observed.Our proposed on-chip temperature controller is expected to enable further miniaturization and multifunctional integration on a single chip for microelectronics.

关键词： Temperature control Low-power electronics On-chip micro temperature controller Freestanding thermoelectric nano films Temperature-sensitive components

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Pulsed THz radiation under ultrafast optical discharge of vacuum photodiode

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Frontiers of Optoelectronics 2024年第2期17卷 99-112页

作者： Aleksandr Ushakov Kseniia Mamaeva Leonid Seleznev Georgy Rizaev Vladimir Bukin Timophey Dolmatov Pavel Chizhov Vladimir Bagdasarov Sergey Garnov Prokhorov General Physics institute of the Russian Academy of Sciences Moscow 119991Russia Physical institute Russian Academy of SciencesMoscow 119991Russia Moscow institute of Physics and Technology Dolgoprudny 141700Russia Russian institute for Scientific and Technical Information Moscow 125190Russia

In this paper,we first present an experimental demonstration of terahertz radiation pulse generation with energy up to 5 pJ under the electron emission during ultrafast optical discharge of a vacuum photodiode.We use a femtosecond optical excitation of metallic copper photocathode for the generation of ultrashort electron bunch and up to 45 kV/cm external electric field for the photo-emitted electron acceleration.Measurements of terahertz pulses energy as a function of emitted charge density,incidence angle of optical radiation and applied electric field have been provided.Spectral and polarization characteristics of generated terahertz pulses have also been studied.The proposed semi-analytical model and simulations in COMSOL Multiphysics prove the experimental data and allow for the optimization of experimental conditions aimed at flexible control of radiation parameters.

关键词： Electron emission Vacuum photodiode Ultrafast excitation Terahertz

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Vaccinia virus viability under different environmental conditions and different disinfectants treatment

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Biosafety and Health 2024年第1期6卷 21-27页

作者： Shan Mei Liang Wei Yu Xie Fei Zhao Yu Huang Zhangling Fan Yamei Hu Liming Wang Lingwa Wang Ying Wang Fengwen Xu Fei Guo NHC Key Laboratory of Systems Biology of Pathogens National Institute of Pathogen Biology and Center for AIDS ResearchChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100730China Department of Otolaryngology Head and Neck Surgery Beijing Tongren HospitalCapital Medical UniversityBejing 100730China Clinical Laboratory Emergency General HospitalBeijing 100028China

Monkeypox(mpox)outbreak in 2022 has caused more than 91,000 cases,has spread to 115 countries,regions,and territories,and has thus attracted much attention.The stability of poxvirus particles in the environment is recognized as an important factor in determining their transmission.However,few studies have investigated the persistence of poxviruses on material surfaces under various environmental conditions,and their sensitivity to biocides.Here,we systematically measured the stability of vaccinia virus(VACV)under different environmental conditions and sensitivity to inactivation methods via plaque assay,quantitative real‐time polymerase chain reaction(qPCR),and Gaussia luciferase(G‐luciferase)reporter system.The results show that VACV is stable on the surface of stainless steel,glass,clothing,plastic,towel,A4 paper,and tissue and persists much longer at 4℃ and?20℃,but is effectively inactivated by ultraviolet(UV)irradiation,heat treatment,and chemical reagents.Our study raises the awareness of long persistence of poxviruses in the environment and provides a simple solution to inactivate poxviruses using common disinfectants,which is expected to help the control and prevention of mpox virus and future poxvirus outbreaks.

关键词： Poxvirus Mpox virus Vaccinia virus Persistence Disinfectant

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Deep Guided Attention Network for Joint Denoising and Demosaicing in Real Image

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Chinese Journal of Electronics 2024年第1期33卷 303-312页

作者： Tao ZHANG Ying FU Jun ZHANG School of Computer Science and Technology Beijing Institute of Technology Advanced Reasearch institute of Multidisciplinary Science Beijing Institute of Technology

Denoising(DN) and demosaicing(DM) are the first crucial stages in the image signal processing pipeline. Recently, researches pay more attention to solve DN and DM in a joint manner, which is an extremely undetermined inverse problem. Existing deep learning methods learn the desired prior on synthetic dataset, which limits the generalization of learned network to the real world data. Moreover, existing methods mainly focus on the raw data property of high green information sampling rate for DM, but occasionally exploit the high intensity and signalto-noise(SNR) of green channel. In this work, a deep guided attention network(DGAN) is presented for real image joint DN and DM(JDD), which considers both high SNR and high sampling rate of green information for DN and DM, respectively. To ease the training and fully exploit the data property of green channel, we first train DN and DM sub-networks sequentially and then learn them jointly, which can alleviate the error accumulation. Besides, in order to support the real image JDD, we collect paired raw clean RGB and noisy mosaic images to conduct a realistic dataset. The experimental results on real JDD dataset show the presented approach performs better than the state-of-the-art methods, in terms of both quantitative metrics and qualitative visualization.

关键词： Image denoising Image demosaicing Joint processing Guided attention Paired real dataset

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