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CHCHD2 Thr61Ile mutation impairs F1F0-ATPase assembly in in vitro and in vivo models of Parkinson's disease

Neural Regeneration Research 2024年第1期19卷 196-204页

作者： Xiang Chen Yuwan Lin Zhiling zhang Yuting Tang Panghai Ye Wei Dai wenlong zhang Hanqun Liu Guoyou Peng Shuxuan Huang Jiewen Qiu Wenyuan Guo Xiaoqin Zhu Zhuohua Wu Yaoyun Kuang Pingyi Xu Miaomiao Zhou Department of Neurology The First Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdong ProvinceChina Department of Neurology The People’s Hospital of Guangxi Zhuang Autonomous RegionNanningGuangxi Zhuang Autonomous RegionChina School of Basic Medical Sciences Guangzhou Medical UniversityGuangzhouGuangdong ProvinceChina Department of Neurology Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

Mitochondrial dysfunction is a significant pathological alte ration that occurs in Parkinson's disease(PD),and the Thr61lle(T61I)mutation in coiled-coil helix coiled-coil helix domain containing 2(CHCHD2),a crucial mitochondrial protein,has been reported to cause Parkinson's disease.FIFO-ATPase participates in the synthesis of cellular adenosine triphosphate(ATP)and plays a central role in mitochondrial energy metabolism.However,the specific roles of wild-type(WT)CHCHD2 and T611-mutant CHCHD2 in regulating F1FO-ATPase activity in Parkinson's disease,as well as whether CHCHD2 or CHCHD2 T61I affects mitochondrial function through regulating F1FO-ATPase activity,remain unclea r.Therefore,in this study,we expressed WT CHCHD2 and T61l-mutant CHCHD2 in an MPP^(+)-induced SH-SY5Y cell model of PD.We found that CHCHD2 protected mitochondria from developing MPP^(+)-induced dysfunction.Under normal conditions,ove rexpression of WT CHCHD2 promoted F1FO-ATPase assembly,while T61I-mutant CHCHD2 appeared to have lost the ability to regulate F1FO-ATPase assembly.In addition,mass spectrometry and immunoprecipitation showed that there was an interaction between CHCHD2 and F1FO-ATPase.Three weeks after transfection with AAV-CHCHD2 T61I,we intraperitoneally injected 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine into mice to establish an animal model of chronic Parkinson's disease and found that exogenous expression of the mutant protein worsened the behavioral deficits and dopaminergic neurodegeneration seen in this model.These findings suggest that WT CHCHD2 can alleviate mitochondrial dysfunction in PD by maintaining F1F0-ATPase structure and function.

关键词： ATP synthase(F1F0-ATPase) coiled-coil helix coiled-coil helix domain containing 2 dopaminergic neuron mitochondrial dysfunction neurodegeneration oligomycin sensitivity-conferring protein Parkinson's disease T61I mutation tyrosine hydroxylase

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Pelagic-benthic coupling of the microbial food web modifies nutrient cycles along a cascade-dammed river

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Frontiers of Environmental Science & Engineering 2022年第4期16卷 125-137页

作者： Nan Yang Linqiong Wang Li Lin Yi Li wenlong zhang Lihua Niu Huanjun zhang Longfei Wang Key Laboratory of Integrated Regulation and Resource Development on Shallow Lake of Ministry of Education College of EnvironmentHohai UniversityNanjing 210098China College of Oceanography Hohai UniversityNanjing 210098China Department of Basin Water Environment Changjiang River Scientific Research InstituteWuhan 430010China Hubei Provincial Key Laboratory of Basin Water Resources and Ecological Environment Sciences Changjiang River Scientific Research InstituteWuhan 430010China

Cascade dams disrupt the river continuum,altering hydrology,biodiversity and nutrient flux.Describing the diversity of multi-trophic microbiota and assessing microbial contributions to the ecosystem processes are prerequisites for the restoration of these aquatic systems.This study investigated the microbial food web structure along a cascade-dammed river,paying special attention to the multi-trophic relationships and the potential role of pelagic-benthic coupling in nutrient cycles.Our results revealed the discontinuity in bacterial and eukaryotic community composition,functional group proportion,as well as a-diversity due to fragmentation by damming.The high microbial dissimilarity along the river,with the total multi-trophic P-diversity was 0.84,was almost completely caused by species replacement.Synchronization among trophic levels suggests potential interactions of the pelagic and the benthic groups,of which the p-diversities were primarily influenced by geographic and environmental factors,respectively.Dam-induced environmental variations,especially hydrological and nutrient variables,potentially influence the microbial food web via both top-down and bottom-up forces.We proposed that the cycles of carbon,nitrogen and phosphorus are influenced by multi-trophic groups through autotrophic and heterotrophic processes,predator-prey relationships,as well as the release of nutrients mainly by microfauna.Our results advance the notion that pelagic-benthic trophic coupling may intensify the accumulation of organic carbon,ammonium and inorganic phosphorus,thereby changing the biogeochemical patterns along river systems.As a consequence,researchers should pay more attention to the multi-trophic studies when assessing the environmental impacts,and to provide the necessary guidance for the ecological conservation and restoration of the dam-regulated systems.

关键词： Reservoir Multi-trophic Beta diversity Predator-prey Nutrient accumulation

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A Case Study on the Rapid Rain-to-Snow Transition in Late Spring 2018 over Northern China:Effects of Return Flows and Topography

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Journal of Meteorological Research 2022年第1期36卷 107-127页

作者： wenlong zhang Xiaopeng CUI Bolong DUAN Bo YU Runxia GUO Haiwen LIU Institute of Urban Meteorology China Meteorological AdministrationBeijing 100089 Key Laboratory of Cloud-Precipitation Physics and Severe Storms Institute of Atmospheric PhysicsChinese Academy of SciencesBeijing 100029 Lanzhou Central Meteorological Observatory Lanzhou 730020 Beijing Weather Forecast Center Beijing 100089 Department of Aviation Meteorology Civil Aviation University of ChinaTianjin 300300 University of Chinese Academy of Sciences Beijing 100049

Phase changes in the precipitation processes of early winter and late spring in midlatitude regions represent challenges when forecasting the timing and magnitude of snowfall.On 4 April 2018,a heavy snow process occurred in Beijing and northwestern Hebei Province,becoming the most delayed occurrence of heavy spring snow ever recorded over Beijing in the last 30 years.This paper uses observational and numerical simulation data to investigate the causes for the rapid rain-to-snow(RRTS)phase transition during this process.The following results are obtained.(1)Return flows(RFs),an interesting type of easterly wind,including those at 1000,925,and 800 hPa,played an important role in this heavy snow process and presented a characteristic"sandwich"structure.The RFs,complex topography,and snow particles that dominated the clouds,were the three key factors for the RRTS transition.(2)The RRTS transition in the plains was directly related to the RF at 925 hPa,which brought about advective cooling initiated approximately 4-6 h before the onset of precipitation.Then,the RF played a role of diabatic cooling when snow particles began to fall at the onset of precipitation.(3)The RRTS transition in the northern part of the Taihang Mountains was closely related to the relatively high altitude that led to a lower surface temperature owing to the vertical temperature lapse rate.Both immediately before and after the onset of precipitation,the snow particles in clouds entrained the middle-level cold air downward,causing the melting layer(from surface to the 0℃-isotherm level)to become very thin;and thus the snow particles did not have adequate time to melt before falling to the ground.(4)The rapid RRTS over the Yanqing mountainous area in the northwest of Beijing could have involved all the three concurrent mechanisms:the advective cooling of RF,the melting cooling of cloud snow particles,and the high-altitude effect.Compared with that in the plain area with less urbanization the duration of the RRTS in the plain area with significant

关键词： midlatitude heavy snow return flow phase transition Bohai Sea cold pool complex terrain numerical simulation

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Preparation and characterization of M_(1)-N_(x)-C_(y)based single atom catalysts for environmental applications

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中国化学快报:英文版 2023年第8期34卷 130-141页

作者： Xinjiang Hu Daixi Zhou Hui Wang wenlong zhang Haoxiang Zhong Yongsheng Chen College of Environmental Science and Engineering Central South University of Forestry and TechnologyChangsha 410004China School of Civil and Environmental Engineering Georgia Institute of TechnologyAtlantaGA 30332United States Georgia Tech Shenzhen Institute Tianjin UniversityShenzhen 518067China

Single atom catalysts(SACs)have become the frontier research fields in catalysis.The M_(1)-N_(x)-C_(y)based SACs,wherein single metal atoms(M1)are stabilized by N-doped carbonaceous materials,have provided new opportunities for catalysis due to their high reactivity,maximized atomic utilization,and high selectivity.In this review,the fabrication methods of M_(1)-N_(x)-C_(y)based SACs via support anchoring strategy and coordination design strategy are summarized to help the readers understand the interaction mechanism of single atoms and support.Then,characterization technologies for identifying single metal atoms are presented.Besides,the environmental applications including management of harmful gases,water purification are discussed.Finally,future opportunities and challenges for preparation strategies,mechanisms and applications are concluded.We conclude this review by emphasizing the fact that M_(1)-N_(x)-C_(y)based SACs has the potential to become an important candidate for solving current and future environmental pollution problems.

关键词： Single atom catalysts Photocatalysis Electrocatalysis Environmental applications

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Crosstalk between IL-15Rα^(+)tumor-associated macrophages and breast cancer cells reduces CD8+T cell recruitment

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Cancer Communications 2022年第6期42卷 536-557页

作者： wenlong zhang Qing zhang Nanfei Yang Qian Shi Huifang Su Tingsheng Lin Zhonglei He Wenxin Wang Hongqian Guo Pingping Shen State Key Laboratory of Pharmaceutical Biotechnology and Department of Urology Nanjing Drum Tower HospitalThe Affiliated Hospital of Nanjing University Medical SchoolSchool of Life SciencesNanjing UniversityNanjingJiangsu 210023P.R.China Department of Urology Drum Tower HospitalMedical School of Nanjing UniversityInstitute of UrologyNanjing UniversityNanjingJiangsu 210008P.R.China Shenzhen Research Institute of Nanjing University Shenzhen 518000China Department of Cellular and Integrative Physiology The University of Texas Health Science Center at San AntonioSan AntonioTexas 78229-3904USA Charles Institute of Dermatology School of MedicineUniversity College DublinDublin Eircode D04 V1W8Ireland

Background:Interleukin-15(IL-15)is a promising immunotherapeutic agent owing to its powerful immune-activating effects.However,the clinical benefits of these treatments are limited.Crosstalk between tumor cells and immune cells plays an important role in immune escape and immunotherapy drug resistance.Herein,this study aimed to obtain in-depth understanding of crosstalk in the tumor microenvironment for providing potential therapeutic strategies to prevent tumor progression.Methods:T-cell killing assays and co-culture models were developed to determine the role of crosstalk between macrophages and tumor cells in breast cancer resistant to IL-15.Western blotting,histological analysis,CRISPR-Cas9 knockout,multi-parameter flow cytometry,and tumor cell-macrophage co-injection mouse models were developed to examine the mechanism by which IL-15Rα^(+)tumor-associated macrophages(TAMs)regulate breast cancer cell resistance to IL-15.Results:We found thatmacrophages contributed to the resistance of tumor cells to IL-15,and tumor cells induced macrophages to express high levels of theαsubunit of the IL-15 receptor(IL-15Rα).Further investigation showed that IL-15Rα^(+)TAMs reduced the protein levels of chemokine CX3C chemokine ligand 1(CX3CL1)in tumor cells to inhibit the recruitment of CD8^(+)T cells by releasing the IL-15/IL-15Rαcomplex(IL-15Rc).Administration of an IL-15Rc blocking peptide markedly suppressed breast tumor growth and overcame the resistance of cancer cells to anti-programmed cell death protein 1(PD-1)antibody immunotherapy.Interestingly,Granulocyte-macrophage colony-stimulating factor(GMCSF)inducedγchain(γc)expression to promote tumor cell-macrophage crosstalk,which facilitated tumor resistance to IL-15.Additionally,we observed that the non-transcriptional regulatory function of hypoxia inducible factor-1alpha(HIF-1α)was essential for IL-15Rc to regulateCX3CL1 expression in tumor cells.Conclusions:The IL-15Rc-HIF-1α-CX3CL1 signaling pathway serves as a crosstalk between macrophages and tumor cells in the t

关键词： breast cancer CD8^(+)T-cell crosstalk IL-15Rα immunotherapy tumor microenvironment tumor-associated macrophages

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Direct reprogramming of fibroblasts into functional hepatocytes via CRISPRa activation of endogenous Gata4 and Foxa3

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中华医学杂志(英文版) 2024年第11期137卷 2024 May 9页

作者： Li Jiacheng Li Ruopu Bai Xue zhang wenlong Nie Yu Hu Shengshou State Key Laboratory of Cardiovascular Disease Fuwai Hospital and Cardiovascular Institute National Center for Cardiovascular Diseases Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China Department of Obstetrics and Gynecology Beijing Advanced Innovation Center for Genomics Third Hospital School of Life Sciences Peking University Beijing China

Background: The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis.Method: The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors—Gata4,Foxa3, andHnf1a—or alternatively, the expression of two transcription factors,Gata4 andFoxa3.In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibroticCol1a1-CreER;Cas9fl/fl mice, effectively activating the expression of endogenousGata4 andFoxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogrammingin vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination.Results: Activation of only two factors,Gata4 andFoxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expandedin vitro and displayed functional characteristics similar to those of mature hepatocyte

关键词： Liver fibrosis CRISPRa Cell reprogramming Transcriptional activation Hepatocytes

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系统论法学视角下的多元解纷机制

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江汉论坛 2024年第1期 118-123页

作者：张文龙华东政法大学数字法治研究院上海201620

当代中国的多元解纷机制不仅从理论走向实践,而且从司法的“权宜之计”发展成为国家治理战略,也为全球治理提供了“中国方案”。当“去司法化”、“司法社会化”日益成为当代中国多元解纷机制发展趋势时,人们迫切需要为多元解纷机制之... 详细信息

当代中国的多元解纷机制不仅从理论走向实践,而且从司法的“权宜之计”发展成为国家治理战略,也为全球治理提供了“中国方案”。当“去司法化”、“司法社会化”日益成为当代中国多元解纷机制发展趋势时,人们迫切需要为多元解纷机制之正当性提供理论基础。多元解纷理论诉诸多元价值来证立其正当性,却陷入悖谬性困境,面临多重悖论。系统论法学观察到,当代中国多元解纷机制之形成是以中国社会的功能分化以及法律系统的分出为基础的。据此,系统论法学认为,随着数字社会的兴起和发展,以数字司法为中心的一站式多元解纷机制,将成为当代中国多元解纷机制的新形态。

关键词：法治中国多元解纷机制功能分化系统论法学

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A numerical implementation of the length-scale independent phase field method

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Acta Mechanica Sinica 2021年第1期37卷 92-104,I0003页

作者： wenlong zhang Ala Tabiei Donald French Department of Civil Engineering University of CincinnatiCincinnatiOH45220USA Department of Mathematics University of CincinnatiCincinnatiOH45220USA

The phase field method for fracture integrates the Griffith theory and damage mechanics approach to predict crack initiation and propagation within one framework.It replaced the discrete representation of crack by diffusive damage and solved it based on a minimization of the global energy storage functional.As a result,no crack tracking topology is needed,and complex crack shapes can be captures without user intervention.However,it is also reported to have an inconsistency between the predicted fracture toughness and the material strength.Recently,a novel energetic degradation function was proposed in literature to handle this issue.This research does some further modifications to the global energy storage functional so that Newton's method can be directly used to solve the energy minimization.With the new energy form,direct implementation of the length-scale independent phase field method into finite element packages like LS-DYNA becomes possible.This paper presents the framework and details of implementing the length-scale independent phase field method into LS-DYNA through a user-defined element and material subroutine.Several numerical examples are presented to compare with the experiment crack shape.Most importantly,this paper is one of the first ones to quantitatively predict accurate force response compared to experiments.These examples verify the accuracy of the new energy form and implementation.

关键词： Phase field method Length-scale independency Newton’s method LS-DYNA

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Characteristics and mechanism of vertical coupling in the genesis of tropical cyclone Durian(2001)

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Science China Earth Sciences 2021年第3期64卷 440-457页

作者： wenlong zhang Xiaopeng CUI Jianxi DONG Institute of Urban Meteorology China Meteorological AdministrationBeijing100089China Key Laboratory of Cloud-Precipitation Physics and Severe Storms Institute of Atmospheric PhysicsChinese Academy of SciencesBeijing100029China National Marine Environmental Forecasting Center Beijing100081China Collaborative Innovation Center on Forecast and Evaluation of Meteorological Disasters Nanjing University of Information Science and TechnologyNanjing210044China University of Chinese Academy of Sciences Beijing100049China

The vertical coupling(VC)process and mechanism during the genesis of a tropical cyclone(TC)implied by the weak vertical shear of horizontal wind,one of the key factors impacting TC genesis,constitute important but unanswered fundamental scientific problems.This paper carried out a targeted investigation of this problem through numerical simulation and theoretical analyses.The main conclusions are as follows.Even if TC genesis occurs in a barotropic environment,a VC process still occurs between the trough(vortex)at the middle level and that at the lower level in the TC embryo area.VC mainly occurs at the tropical disturbance(TDS)stage.Only after the VC is accomplished can the tropical depression(TD)organize further by itself and develop into the tropical storm(TS)stage or the stronger tropical typhoon(TY)stage through the WISHE(wind-induced surface heat exchange)mechanism.In the VC process,vortical hot towers(VHTs)play vertical connecting roles and are the actual practitioners of the VC.Through the VHTs’vertical connections,the middle-and lower-troposphere trough axes move towards each other and realize the VC.VHTs can produce intensive cyclonic vorticity in the lower troposphere,which is mainly contributed by the stretching term.The tilting term can produce a single dipole or double dipole of vorticity,but the positive and negative vorticity pairs offset each other roughly.While the stretching term ensures that the cyclonic rotations of the wind field in the middle and lower levels tend to be consistent,the tilting term acts to uniformly distribute the horizontal wind in the vertical direction,and both terms facilitate the VC of the wind field.With the latent heat of condensation,VHTs heat the upper and middle troposphere so that the 352 K equivalent potential temperature contour penetrates vertically into the 925–300 hPa layer,realizing the VC of the temperature field.While forming cloud towers,VHTs make the ambient air become moist and nearly saturated so that the 95%relative humidity contour penetrates vertica

关键词： Genesis of tropical cyclones Vertical coupling Vortical hot tower Axisymmetric distribution Barotropic instability

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Effect of Tb(Ⅲ) on the unfolding of ciliate Euplotes octocarinatus centrin induced by guanidine hydrochloride

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Journal of Rare Earths 2018年第2期36卷 203-208页

作者： Enxian Shi wenlong zhang Binsheng Yang Institute of Molecular Science Shanxi University Department of Pharmacy Shanxi Medical University

To understand the unfolding of ciliate Euplotes octocarinatus centrin（EoCen）, the glycine positioned at 115, the sixth residue of the loop of the protein＇s third EF-hand, was mutated into tryptophan（Trp）.Intrinsic fluorescence and Tb（Ⅲ） binding properties of wild type EoCen and G115W mutant were monitored by fluorescence spectra in 10 mmol/L Hepes. The emission maximum of EoCen was 306 nm and mutation had no impact on the Tb（Ⅲ） binding properties. The properties of G115W were investigated by fluorescence, far-UV circular dichroism（CD） spectra and fluorescence decays in the absence or in the presence of 6 mol/L guanidine hydrochloride（GdnHCl）. For the increase in polarity of microenvironment around Trp residue, the emission maximum of apoG115 W at 343 nm is shifted to 359 nm in 6 mol/L GdnHCI. Also the secondary structure is lost nearly and fluorescence lifetime decreases in 6 mol/L GdnHCI. The unfolding of G115W induced by GdnHCI was assessed by using the model of structural element The unfolding of proteins is a sequential reaction, namely two-transition. three-state process. The first transition belongs to the unfolding of the C-terminal domain, and the second transition is assigned to the unfolding of the N-terminal domain. The ⊿〈△G_（total）~0（H_2O）〉 was used to determine the effect of Tb（Ⅲ） on the stability of apoprotein. The 〈△G_（total）~0（H_2 O）〉for Tb_2-G115 W has a less increase of0.68 kJ/mol compared with apoG115W, proving Tb（Ⅲ） situated at C-terminal has negligible impact on the stability of protein. Whereas the 〈△G_（total）~0（H_2 O）〉 for Tb_4-G115W has a rise of 1.29 kJ/mol compared with Tb_2-G115W, manifesting Tb（Ⅲ） lcocated at low affinity sites has considerable influence on protein stability. mainly stabilizing the N-terminal domain.

关键词： Euplotes octocarinatus centrin Tb（Ⅲ） Unfolding Structure element Stability Rare earths

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