Intravital lipid droplet labeling and imaging reveals the phenotypes and functions of individual macrophages in vivo

Macrophages play pivotal roles in the maintenance of tissue homeostasis. However, the reactivation of macrophages toward proinflammatory states correlates with a plethora of inflammatory diseases, including atherosclerosis, obesity, neurodegeneration, and bone marrow (BM) failure syndromes. The lack of methods to reveal macrophage phenotype and function in vivo impedes the translational research of these diseases. Here, we found that proinflammatory macrophages accumulate intracellular lipid droplets (LDs) relative to resting or noninflammatory macrophages both in vitro and in vivo, indicating that LD accumulation serves as a structural biomarker for macrophage phenotyping. To realize the staining and imaging of macrophage LDs in vivo, we developed a fluorescent fatty acid analog-loaded poly(lactic-co-glycolic acid) nanoparticle to label macrophages in mice with high efficiency and specificity. Using these novel nanoparticles, we achieved in situ functional identification of single macrophages in BM, liver, lung, and adipose tissues under conditions of acute or chronic inflammation. Moreover, with this intravital imaging platform, we further realized in vivo phenotyping of individual macrophages in the calvarial BM of mice under systemic inflammation. In conclusion, we established an efficient in vivo LD labeling and imaging system for single macrophage phenotyping, which will aid in the development of diagnostics and therapeutic monitoring. Moreover, this method also provides new avenues for the study of lipid trafficking and dynamics in vivo.Supplementary key words: macrophage, inflammation, lipid droplet, nanoparticle delivery, in vivo imaging, fatty acid analog, bone marrow, systemic inflammation, lipid trafficking, biomarker

Macrophages, a type of immune cells, almost reside in all tissues of body, from the skin to the bone marrow (BM) (1). Macrophages have remarkable plasticity, and they can be activated into specific subtypes by modifying their physiology and functions in response to local environmental cues. Activated macrophages are commonly divided into proinflammatory killing subtype and anti-inflammatory repairing subtype. Proinflammatory macrophages responding to bacteria, IFN-γ, and lipopolysaccharide (LPS) are involved in host defense and inflammation, whereas anti-inflammatory macrophages responding to interleukin-4 (IL-4), IL-10, and IL-13 play a pivotal role in tissue homeostasis and remodeling (2). Increasing evidence indicates that the reactivation of macrophages toward proinflammatory states under diverse kinds of stress is correlated with a plethora of inflammatory diseases, such as atherosclerosis, diabetes, obesity, rheumatoid arthritis, neurodegeneration, and BM failure syndromes (3, 4). Thus, characterization of macrophage activation status and the underlying molecular mechanism in situ will help elucidate their functions in these diseases; however, in vivo analysis of the macrophage activation status in their native multicellular microenvironment is challenging.Although lipid droplets (LDs) have been initially described as intracellular fat storage organelles in adipocytes, increasing studies indicate that myeloid cells also form LDs under inflammation and stress (5, 6). Macrophages, as the effector cells of innate immunity, are found to form LDs to support their host defense when exposed to pathogens, such as parasites, bacteria, and viruses (7, 8, 9, 10, 11). However, abnormal LD accumulation in tissue-resident macrophages correlates with the pathogenesis of various inflammatory diseases. For instance, foam cells in atherosclerotic lesions can maintain the local inflammatory response by secreting proinflammatory cytokines (12, 13, 14). Moreover, LD-accumulating microglia contribute to neurodegeneration by producing high levels of reactive oxygen species (ROS) and secreting proinflammatory cytokines (15). These findings indicate that LD accumulation might be a hallmark of macrophages with proinflammatory functions.In this study, based on the typical activation of in vitro BM-derived macrophages, we find that proinflammatory M_{(LPS + IFN-γ)} macrophages are characterized by LD accumulation, whereas resting macrophages and anti-inflammatory M_(IL-4) and M_(IL-10) macrophages do not contain any LDs. These features also hold for Matrigel plug-recruited macrophages and tissue-resident macrophages in mice. These findings demonstrate that LD accumulation could serve as a morphological index to distinguish proinflammatory macrophages from others.It is feasible to distinguish LD-containing cells using imaging techniques, which has translational potential for identification of proinflammatory macrophages in vivo. However, current techniques for LD visualization are traditional in vitro staining method, and in vivo staining and imaging of LD in individual macrophages remains a challenge. Through nanocarrier screening, we selected the poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) as nanocarrier to deliver the lipophilic carbocyanine dye (DiI_C18(5) solid (1,1''-dioctadecyl-3,3,3'',3''-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt) [DiD]) and lipid staining dye (C1-BODIPY 500/510-C12) into macrophages. Using these dual fluorescence-labeled PLGA NPs, we achieved in situ and in vivo functional identification of single macrophages in various tissues under systemic or local inflammatory stress. Collectively, this study establishes an efficient in vivo labeling and imaging system of intracellular LDs for phenotyping the activation status and functions of individual macrophages in their dynamic niche, which is pivotal for disease diagnosis and preclinical research.     

城市可持续规划中的生态敏感区避让分析——以丽江市为例

为促进城市可持续发展,在科学合理规划城市空间格局的基础上,应突出强调保护原有特色生态环境要素,保育生态系统服务功能和原住民人居环境。作为世界文化遗产,丽江城市面貌在过去数十年发生了深刻变化,城市用地显著扩张,同时严峻的环境问题对城市可持续发展提出挑战。以丽江市为例,选取对其发展影响深远的水系元素、噪声环境因子和森林火灾3个关键生态敏感因子,提出通过分析这类生态敏感区在城市规划过程中的避让方法、程度,区划城市规划基本空间格局,以避免城市未来因生态环境问题突出而制约其发展的规划用地思路。结果表明,丽江城区周边湖泊水系、古城内部及周边原住民人口密集区都是保障城市可持续发展的生态保护重点地区,在城市用地规划中必须予以避让;该区域面积为147.2 km2,占研究区面积的11.1%。同时,明确给出了规划用地的具体空间控制范围和相应的避让原则。     

鱼类生物群落对太子河流域土地利用、河岸带栖息地质量的响应

河流生态系统的退化是多空间尺度环境因子作用的结果。探讨不同尺度环境因子及水生生物之间的作用关系,识别影响水生生物群落完整性的尺度问题,是有效开展水生生物保护的基础。基于2009年对太子河流域15个样点的鱼类、河岸带栖息地质量评价,结合遥感影像解译的太子河流域土地利用情况(包括流域尺度和河段尺度),研究鱼类完整性指数(F-IBI)与两种尺度土地利用、栖息地质量参数之间的关系。结果表明太子河上游地区河岸栖息地质量较好,下游地区由于农业用地、城镇用地比例的增加河岸栖息地质量明显下降。F-IBI与自然用地比例呈正相关,与农业、城镇用地比例呈负相关。农业用地对F-IBI的影响体现在流域尺度,而城镇用地在两种尺度上都存在显著影响。相比于农业用地,城镇用地相同比例的增加会导致F-IBI更快的下降。底质、水质状况、人类活动强度是显著影响F-IBI的栖息地质量评价参数。3项参数均随农业和城镇用地比例增加而降低,农业用地主要在流域尺度上对3项参数产生影响,城镇用地主要影响底质和水质状况2项参数,而在两种尺度上的影响相差不大。     

中国电力行业CO2减排潜力及其贡献因素

电力行业低碳转型是中国低碳经济转型进程中关键行业之一,如何科学分析电力行业的碳减排潜力,确定操作性强的低碳转型路线、提出有效的政策措施是中国政府亟待解决的焦点问题之一。考虑终端电力消费、低碳能源发电占比、火力发电结构、火力发电效率、线损率等因素,构建了自底向上的电力行业CO2排放核算模型,在此基础上,利用情景分析方法探索中国电力行业2015和2020年的CO2减排潜力,进一步利用对数平均权重分解法(LMDI,Logarithmic Mean weight Divisia Index method)对电力行业CO2减排影响因素的贡献度做了归因分析。结果显示,相比基准情景,在当前政策情景和低碳政策情景下,电力行业将分别带来27.0亿t和36.9亿t的CO2减排量。低碳能源发电和火力发电效率是未来对CO2减排最重要的两个贡献因素。终端电力消费量一直是促进电力行业CO2排放增长最重要的贡献因素,因此通过电力需求侧管理等手段控制电力消费量对电力行业的低碳发展至关重要。最后结合减排贡献因素分析的结果为中国电力行业低碳发展提出了相应的政策建议。     

石灰石粉施用剂量对重庆酸雨区受害 马尾松林细根生长的影响

马尾松(Pinus massoniana)为重庆的重要乡土树种和主要造林树种,担负着特殊而重要的生态环境保全功能,但在酸雨的长期影响下,其健康状况持续低下。为探究不同剂量石灰石粉对马尾松细根生长的影响,给受害马尾松纯林的恢复与管理提供科学依据,2004年5月在属于酸雨区的重庆市江北区铁山坪林场,采用随机区组设计,设立在马尾松纯林酸化土壤表面一次性撒施石灰石粉0(不撒施对照)、1、2、3,4t/hm² 5个处理的长期定位观测试验。2006年10月的监测结果表明,与对照相比,撒施石灰石粉可明显促进腐殖质层和0-20、20-40、40-60cm土层马尾松细根的生长(P < 0.05),尤其是改善腐殖质层和20-40、40-60cm的根量,在腐殖质层和0-60cm马尾松细根的干重总密度、长度总密度、表面积总密度、体积总密度和根尖总密度与对照各项密度指标对应值的比值,1t/hm²下分别为1.53、1.34、1.39、1.24,1.48,2t/hm²下分别为2.05、1.83、1.94、1.79,2.04,3t/hm²下分别为2.17、2.02、2.02、1.78,2.32,4t/hm²下分别为2.68、2.52、2.62、3.70和2.71,改善效果总体上随撒施剂量增大而增强。从有限时间(2-3a)内观测的根系生长改善效果与经济投入的比值来看,建议一次性撒施的最佳剂量为2t/hm²,但在考虑时间加长情况下,效果最好和最经济的剂量还有待后续研究。     

50年来山东塔山植被与物种多样性的变化

为分析塔山植被与物种多样性50a自然演替和动态变化,2009-2010年,采用系统勘踏法和典型取样法进行了调查。当前塔山主要植被类型为"黑松林-赤松林-栓皮栎林"。50a间,该区针叶林从黑松(Pinus thunbergii)林演替为黑松林、赤松(P. densiflora)林和油松林(P. tabulaeformis)为主的混合针叶林,但针叶林的整体优势度下降,以栓皮栎(Quercus variabilis)和麻栎(Q. acutissima)为建群种的阔叶林面积明显增大,由针叶林向阔叶林的演替趋势明朗。物种丰富度为草本层>灌木层>乔木层,Shannon-wiener指数和Simpson指数为灌木层>草本层>乔木层,物种多样性较低,处于森林演替初期。     

Depression compromises antiviral innate immunity via the AVP-AHI1-Tyk2 axis

Depression is a serious public-health issue. Recent reports have suggested higher susceptibility to viral infections in depressive patients. However, how depression affects antiviral innate immune signaling remains unknown. Here, we revealed a reduction in expression of Abelson helper integration site 1 (AHI1) in the peripheral blood mononuclear cells (PBMCs) and macrophages from the patients with major depressive disorder (MDD), which leads to attenuated antiviral immune response. We found that depression-related arginine vasopressin (AVP) induces reduction of AHI1 in macrophages. Further studies demonstrated that AHI1 is a critical stabilizer of basal type-I-interferon (IFN-I) signaling. Mechanistically, AHI1 recruits OTUD1 to deubiquitinate and stabilize Tyk2, while AHI1 reduction downregulates Tyk2 and IFN-I signaling activity in macrophages from both MDD patients and depression model mice. Interestingly, we identified a clinical analgesic meptazinol that effectively stimulates AHI1 expression, thus enhancing IFN-I antiviral defense in depression model mice. Our study promotes the understanding of the signaling mechanisms of depression-mediated antiviral immune dysfunction, and reveals meptazinol as an enhancer of antiviral innate immunity in depressive patients.Subject terms: Innate immunity, Ubiquitylation, Cell signalling     

基于LMDI分解的厦门市碳排放强度影响因素分析

研究碳排放强度的变化趋势及其影响因素对于指导低碳城市建设具有重要意义。应用对数平均权重分解法(LMDI),基于厦门市2005—2010年各部门终端消费数据对碳排放强度指标进行因素分解,并将传统分析仅注重产业部门的能源碳排放,拓展到全面考虑产业部门和家庭消费的能源活动和非能源活动影响。研究结果表明:2005—2010年厦门市碳排放强度下降17.29%,其中产业部门能源强度对总碳排放强度变化影响最大(贡献63.07%),家庭消费能源强度是碳排放强度下降的主要抑制因素(-45.46%)。从影响效应角度看,经济效率对碳排放强度下降贡献最大,碳排系数减排贡献最小;从部门减排贡献角度看,第二产业贡献最大,家庭消费贡献最小。总体而言,厦门市未来碳减排重点部门在第二产业,优化产业结构和能源结构有较大减排潜力。     

西江下游浮游植物群落周年变化模式

西江肇庆段是珠江干流汇入珠三角河网水域的咽喉通道,对2009年该江段的浮游植物群落组成及变化进行系统阐析.调查期间共发现浮游植物7门,245种(包括变种、变型),其中硅藻和绿藻是浮游植物群落组成的最主要类群,分别占总种数的42.44％和34.69％,其次是裸藻和蓝藻.PCA分析结果显示,浮游植物各类群的种类丰富度和生物量的周年变化趋势主要受水温和径流量的影响.浮游植物种类丰富度的周年变化呈现明显的高温季节高,低温季节低的特征,除因大多数藻类物种直接喜好高温之外,径流量的增大有助于真浮游绿藻种类的外源注入及半浮游和偶然性浮游硅藻种类的增加.浮游植物生物量的周年变化呈现明显的双峰型,峰值分别出现在8月和11月.然而8月份第1个高峰出现之前,生物量波动不大,尽管水温的升高有助于生物量的增加,但是径流量增大所带来的稀释作用掩盖了水温上升对浮游植物生长的促进作用.真浮游植物生物量的变化趋势和数值与总浮游植物极其接近,主要得益于真浮游硅藻物种颗粒直链藻在总种群中的绝对优势地位.综上,水温升高对浮游植物种类丰富度和生物量的增长均有促进,径流量的增大虽然有助于种类丰富度的增加,但不利于生物量的增加.