Comparative Analysis of Platelet-Derived Extracellular Vesicles Using Flow Cytometry and Nanoparticle Tracking Analysis

Growing interest in extracellular vesicles (EVs) has prompted the advancements of protocols for improved EV characterization. As a high-throughput, multi-parameter, and single particle technique, flow cytometry is widely used for EV characterization. The comparison of data on EV concentration, however, is hindered by the lack of standardization between different protocols and instruments. Here, we quantified EV counts of platelet-derived EVs, using two flow cytometers (Gallios and CytoFLEX LX) and nanoparticle tracking analysis (NTA). Phosphatidylserine-exposing EVs were identified by labelling with lactadherin (LA). Calibration with silica-based fluorescent beads showed detection limits of 300 nm and 150 nm for Gallios and CytoFLEX LX, respectively. Accordingly, CytoFLEX LX yielded 40-fold higher EV counts and 13-fold higher counts of LA⁺CD41⁺ EVs compared to Gallios. NTA in fluorescence mode (F-NTA) demonstrated that only 9.5% of all vesicles detected in scatter mode exposed phosphatidylserine, resulting in good agreement of LA⁺ EVs for CytoFLEX LX and F-NTA. Since certain functional characteristics, such as the exposure of pro-coagulant phosphatidylserine, are not equally displayed across the entire EV size range, our study highlights the necessity of indicating the size range of EVs detected with a given approach along with the EV concentration to support the comparability between different studies.     

Residual attention-based tracking-by-detection network with attention-driven data augmentation

Tracking-by-detection (TBD) is a significant framework for visual object tracking. However, current trackers are usually updated online based on random sampling with a probability distribution. The performance of the learning-based TBD trackers is limited by the lack of discriminative features, especially when the background is full of semantic distractors. We propose an attention-driven data augmentation method, in which a residual attention mechanism is integrated into the TBD tracking network as supplementary references to identify discriminative image features. A mask generating network is used to simulate changes in target appearances to obtain positive samples, where attention information and image features are combined to identify discriminative features. In addition, we propose a method for mining hard negative samples, which searches for semantic distractors with the response of the attention module. The experiments on the OTB2015, UAV123, and LaSOT benchmarks show that this method achieves competitive performance in terms of accuracy and robustness.     

In Vivo Electrophysiology of Peptidergic Neurons in Deep Layers of the Lumbar Spinal Cord after Optogenetic Stimulation of Hypothalamic Paraventricular Oxytocin Neurons in Rats

The spinal ejaculation generator (SEG) is located in the central gray (lamina X) of the rat lumbar spinal cord and plays a pivotal role in the ejaculatory reflex. We recently reported that SEG neurons express the oxytocin receptor and are activated by oxytocin projections from the paraventricular nucleus of hypothalamus (PVH). However, it is unknown whether the SEG responds to oxytocin in vivo. In this study, we analyzed the characteristics of the brain–spinal cord neural circuit that controls male sexual function using a newly developed in vivo electrophysiological technique. Optogenetic stimulation of the PVH of rats expressing channel rhodopsin under the oxytocin receptor promoter increased the spontaneous firing of most lamina X SEG neurons. This is the first demonstration of the in vivo electrical response from the deeper (lamina X) neurons in the spinal cord. Furthermore, we succeeded in the in vivo whole-cell recordings of lamina X neurons. In vivo whole-cell recordings may reveal the features of lamina X SEG neurons, including differences in neurotransmitters and response to stimulation. Taken together, these results suggest that in vivo electrophysiological stimulation can elucidate the neurophysiological response of a variety of spinal neurons during male sexual behavior.     

In vivo Targeting of DNA Vaccines to Dendritic Cells via the Mannose Receptor Induces Long-Lasting Immunity against Melanoma

Herein, we report effective, C-type lectin mannose receptor (MR)-selective, in vivo dendritic cell (DC)-targeting lipid nanoparticles (LNPs) of a novel lipid-containing mannose-mimicking di-shikimoyl- and guanidine head group and two n-hexadecyl hydrophobic tails (DSG). Subcutaneous administration of LNPs of the DSG/p-CMV-GFP complex showed a significant expression of green fluorescence protein in the CD11c⁺ DCs of the neighboring lymph nodes compared to the control LNPs of the BBG/p-CMV-GFP complex. Mannose receptor-facilitated in vivo DC-targeted vaccination (s.c.) with the electrostatic complex of LNPs of DSG/pCMV-MART1 stimulated long-lasting (270 days post B16F10 tumor challenge) antimelanoma immunity under prophylactic conditions. Remarkably, under therapeutic settings, vaccination (s.c.) with LNPs of the DSG/pCMV-MART1 complex significantly delayed melanoma growth and improved the survival of mice with melanoma. These findings demonstrate that this nonviral delivery system offers a resilient and potential approach to deliver DNA vaccines encoding tumor antigens to DCs in vivo with high efficacy.     

气藏平均地层压力跟踪计算新方法

平均地层压力是产能评价和动态分析的基础，准确、快速获取平均地层压力对高效开发气藏意义重大。基于地层压力随时间变化的规律，分析了平均地层压力的变化规律。研究结果表明：平均地层压力等效点仅随时间发生改变，平均地层压力的下降速率等于或者近似等于井底流压的下降速率。从封闭弹性驱动气藏的物质平衡方程出发，考虑偏差系数和井底流压随平均地层压力的变化，推导建立了平均地层压力跟踪计算新方法，根据生产数据可迭代计算平均地层压力。方法验证结果显示，采气速度和采出程度共同影响模型的计算结果。应用实例表明，跟踪计算法与压力恢复试井和物质平衡法之间的相对误差均较小，满足工程计算精度要求，且跟踪计算法不需依托生产测试数据，节约了测试费用，避免了测试占产。     

Direct View on the Origin of High Li+ Transfer Impedance in All-Solid-State Battery

Large interfacial resistance plays a dominant role in the performance of all-solid-state lithium-ion batteries. However, the mechanism of interfacial resistance has been under debate. Here, the Li⁺ transport at the interfacial region is investigated to reveal the origin of the high Li⁺ transfer impedance in a LiCoO₂(LCO)/LiPON/Pt all-solid-state battery. Both an unexpected nanocrystalline layer and a structurally disordered transition layer are discovered to be inherent to the LCO/LiPON interface. Under electrochemical conditions, the nanocrystalline layer with insufficient electrochemical stability leads to the introduction of voids during electrochemical cycles, which is the origin of the high Li⁺ transfer impedance at solid electrolyte-electrode interfaces. In addition, at relatively low temperatures, the oxygen vacancies migration in the transition layer results in the formation of Co₃O₄ nanocrystalline layer with nanovoids, which contributes to the high Li⁺ transfer impedance. This work sheds light on the mechanism for the high interfacial resistance and promotes overcoming the interfacial issues in all-solid-state batteries.     

伊拉克A油田Asmari组混积型碳酸盐岩储层隔夹层研究

伊拉克A油田Asmari组为大型混积型碳酸盐岩油藏,发育灰岩、白云岩、砂岩、泥岩和硬石膏等多种岩性,储层复杂、非均质性强,存在多种类型的隔夹层。基于岩心、薄片、测井和分析测试等资料,对研究区隔夹层类型及特征展开综合分析,识别出了泥岩、硬膏岩和泥-微晶碳酸盐岩3类隔夹层。泥岩和硬石膏致密无孔隙、基本不具有渗流能力,对流体流动可以起到阻挡作用;泥-微晶碳酸盐岩属于细粒碳酸盐岩沉积,孔隙度低,但一般裂缝较发育,具有一定渗流能力,不能对流体起到有效的隔挡作用。硬石膏主要在油田中部及北部A段发育,其对油田北部储层纵向连通起到一定隔挡作用,导致纵向压力衰竭不一致。泥岩主要在C段顶部、B4和B1层发育,平面上主要在油田南部较发育。C段顶部发育一套厚度较大、稳定分布的泥岩隔层,将C段和上部B段及A段分隔为两个独立的油水系统,并有效阻挡了C段水层锥进到B段;B4和B1层泥岩较发育,但由于油田南部区域断层和裂缝较发育,导致泥岩隔夹层未能起到有效隔挡作用。搞清研究区隔夹层类型、分布及是否起隔挡作用,对A油田各区域各小层有效动用开发、见水规律研究具有很好的指导意义。     

Hydrodynamics study of a fast liquid–liquid oxidation process with in situ gas production in microreactors

Hydrodynamics characteristics of a fast and highly exothermic liquid–liquid oxidation process with in situ gas production in microreactors were studied using a newly developed experimental method. In the adipic acid synthesis through the K/A oil (the mixture of cyclohexanol and cyclohexanone) oxidation with nitric acid, bubble generation modes were divided into four categories. The gas production became more intensive, unstable, even explosive with increasing the oil phase feed rate and the temperature. A novel automatic image processing method was developed to monitor the instantaneous velocity online by tracking the gas–liquid interface. The axial velocity at the same location was unstable due to the changing gas production rate. Furthermore, the actual residence time was obtained easily with being only 36% of the space–time minimally, beneficial for establishing accurate kinetics and mass transfer models with time participation. Finally, an empirical correlation was developed to predict the actual residence time under different conditions.     

基于现代审美下的赣傩艺术元素分析与设计应用

目的 赣傩艺术元素内容丰富并保存完整,是研究我国古代赣傩地区文化与艺术的珍贵史料.研究赣傩艺术元素的提取及衍生图案的设计方法,在设计应用中表达赣傩傩面具、选取傩舞形和构筑傩庙魂,能给当代赣傩艺术元素的产品设计以指引.方法 通过直接引用、形意结合和局部强化的设计方法,增厚其丰富内涵、设计个性和主题意蕴.通过现代化审美中"绿色化、感性化、地域化"的要求,遵循"以退为进,化繁为简""多元并存,个性体现"的原则,强化赣傩艺术元素的独特风格.结论 用现代的手法,将特色的赣傩艺术元素植入到理性的产品设计中去,使赣傩艺术元素与现代设计思想有机结合.在继承传统文化与现代流行中找到契合点,深入挖掘赣傩艺术元素蕴含的文化精神、文化胸怀和文化自信,使中华传统文化在现代审美下进行创新再现.     

农耕印迹:扎染菱形纹样作为布依语“Ndaeh Sengh”的解读研究

目的 菱形纹样是我国古老的几何纹样之一，其历史悠久、应用广泛，在不同地域和文化背景下被赋予了不同的文化内涵，厘清纹样的在地性符号所指，是提升传统工艺传承与设计创新的方法路径。方法 以贵州花戛乡布依族扎染技艺中的菱形纹样为研究个案，结合人类学的田野调查法和艺术设计学的比较研究法，对花戛布依族服装配饰中的扎染菱形纹样进行综合考证和分析。结论 认为花戛扎染菱形纹样是布依族日用器物“升”的形态转译，是农耕背景下布依人祈求丰收的文化符号载体。有着文化符号功能的菱形“Ndaeh Sengh”纹样，与布依人生活、习俗、地域环境等方面存在多维的联结，“Ndaeh Sengh”越多代表农作物收成越好，寓意丰收和富足。“Ndaeh Sengh”作为农耕记忆背景下布依族集体无意识原型形式的外化，其制作工艺和纹样所承载的文化记忆对于非遗保护、工艺传承、非遗设计等方面都有一定的理论指导意义。