香蕉枯萎病菌内源报告基因Foc4carS的鉴定及其应用

香蕉枯萎病是由尖孢镰刀菌古巴转化型(Fusarium oxysporum f. sp. cubense, Foc)引起的香蕉毁灭性土传病害,其中4号生理小种(Foc4)能感染几乎所有的香蕉品系,危害最严重。carS基因通过调控下游car结构基因参与调控镰刀菌类胡萝卜素的生物合成,本研究克隆鉴定了Foc4carS基因(FOIG_05085),Foc4carS蛋白具有典型的RING-finger蛋白结构域。利用分割标记法(Split-marker PCR)获得Foc4carS基因的融合片段,同时构建含有Foc4carS基因sgRNA591序列的pUC-fFuCas9-HTBNLS-hph-Foc4carS基因编辑载体,通过PEG介导的原生质体转化获得该基因的敲除突变体、回补突变体以及基因编辑敲除体,并对敲除和回补突变体的生物学特性和致病力进行分析。结果显示:ΔFoc4carS突变体的菌落直径、产孢量和致病力等生物学表型与野生菌株Foc4无显著差异,而ΔFoc4carS突变体菌落颜色呈深橙色,Foc4carS基因的缺失影响了次生代谢产物类胡萝卜素的生物合成;基因编辑的ΔFoc4carS(HDR)突变体不论是再生筛选板还是继代后的PDA平板,其菌落均出现典型的深橙色,表明Foc4carS可作为内源报告基因,在香蕉枯萎菌Foc4中进行基因质粒型CRISPR/Cas9编辑可行。

Freight Vehicle Routing Optimization for Sporadic Orders Using Floating Car Data

The increasing popularity of e-commerce brings large volumes of sporadic orders from different customers,which have to be handled by freight trucks and distribution centers. To improve the level of service and reduce the total shipping cost as well as traffic congestions in urban area, flexible methods and optimal vehicle routing strategies should be adopted to improve the efficiency of distribution effort. An optimization solution for vehicle routing and scheduling problem with time window for sporadic orders (VRPTW- S) was provided based on time-dependent travel time extracted from floating car data (FCD) with ArcGIS platform. A VRPTW-S model derived from the traditional vehicle routing problem was proposed, in which uncertainty of customer orders and travel time were considered. Based on this model, an advanced vehicle routing algorithm was designed to solve the problem. A case study of Shenzhen, Guangdong province, China, was conducted to demonstrate the vehicle operation flow,in which process of FCD and efficiency of delivery systems under different situations were discussed. The final results demonstrated a good performance of application of time-dependent travel time information using FCD in solving vehicle routing problems.

碧根果致敏原Car i 1的分离纯化及表征鉴定

目的分离纯化碧根果致敏原Car i 1,并对其结构进行表征鉴定。方法以新鲜碧根果果仁为原料,通过粉碎、脱脂、浸提、粗分级、凝胶过滤层析,对碧根果致敏原蛋白Car i 1进行分离纯化。结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱法和免疫印迹法3种方法对Cari1进行鉴定,并通过圆二色谱仪与紫外分光光度计表征其二、三级结构。结果本方法纯化获得碧根果致敏原Cari1,单轮制备量可达5 mg以上,且纯度大于95%,蛋白质高级结构未被破坏,能够被全部3名碧根果过敏患者的血清准确识别。结论该纯化方法技术路线简单、设备要求低且单次制备量高,总得率可达65%,操作便捷,为碧根果致敏原Car i 1的相关研究奠定了物质基础。

基于ADAMS/Car的商用车平顺性仿真分析

为分析随机和脉冲路面对商用车行驶平顺性的影响,以某商用车为研究对象,依托多体动力学理论,使用ADAMS/Car软件构建包含前后悬架、轮胎、车身、转向系统及人体-座椅系统的车辆动力学模型,并对车辆进行了脉冲输入和随机输入平顺性分析,仿真结果表明:在脉冲路面的激励下,车速提高后车辆行驶平顺性提高;随机路面输入条件下,相较于B级路面,C级路面对平顺性的影响更显著,在商用车行驶舒适性评估和改进中应用更为有效。

生酮饮食疗法对孤独症患儿症状及ATEC和CARS评分的改善效果

目的 探讨生酮饮食疗法对孤独症患儿症状及孤独症治疗评估量表(ATEC)和及儿童孤独症评定量表(CARS)评分的改善效果。方法 回顾性选取2021年7月至2022年7月收治的78例ASD患儿为研究对象,根据不同治疗方案将其分为对照组和治疗组,每组39例,对照组接受ASD特殊教育及日常训练+普通饮食,治疗组接受ASD特殊教育及日常训练+生酮饮食,均连续治疗6个月,比较2组《自闭症儿童心理教育评核(第三版)》(C-PEP-3)评分、孤独症治疗评估量表(ATEC)评分、儿童孤独症评定量表(CARS)评分及孤独症行为量表(ABC)评分。结果 治疗后,2组C-PEP-3各维度评分及总分均显著上升,且治疗组高于对照组(P<0.05);治疗后,2组ATEC各维度(表达/语言沟通、社交能力、感知/认知能力及健康/生理/行为)评分均显著下降,且治疗组低于对照组(P<0.05);治疗后,2组CARS和ABC评分均显著下降,且治疗组低于对照组(P<0.05);治疗后,研究组就寝习惯不良、睡眠焦虑、睡眠持续时间不规律、白天嗜睡、夜醒、入睡潜伏期延长及CSHQ总分均显著低于对照组(P<0.05)。结论 生酮饮食疗法可有效控制孤独症患儿临床症状,改善其认知及异常行为,降低其疾病程度,提高其睡眠质量,值得临床推广。

Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) via dephosphorylation of the EGFR signaling pathway

The current study aimed to assess the effect of timosaponin AⅢ(T-AⅢ)on drug-metabolizing enzymes during anticancer therapy.The in vivo experiments were conducted on nude and ICR mice.Following a 24-day administration of T-AⅢ,the nude mice exhibited an induction of CYP2B10,MDR1,and CYP3A11 expression in the liver tissues.In the ICR mice,the expression levels of CYP2B10 and MDR1 increased after a three-day T-AⅢ administration.The in vitro assessments with HepG2 cells revealed that T-AⅢ induced the expression of CYP2B6,MDR1,and CYP3A4,along with constitutive androstane receptor(CAR)activation.Treatment with CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4 expression.Furthermore,other CAR target genes also showed a significant increase in the expression.The up-regulation of murine CAR was observed in the liver tissues of both nude and ICR mice.Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation,with this effect being partially reversed by the ERK activator t-BHQ.Inhibition of the ERK1/2 signaling pathway was also observed in vivo.Additionally,T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845,and suppressed EGF-induced phosphorylation of EGFR,ERK,and CAR.In the nude mice,T-AⅢ also inhibited EGFR phosphorylation.These results collectively indicate that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.

CAR-T疗法安全性控制技术专利现状

CAR-T疗法属于先进细胞治疗手段,自问世以来,因其在肿瘤治疗中发挥的卓越的治疗效果而备受关注,但由于受制于临床治疗过程的各种毒副作用,这一疗法的广泛推广仍然还有很长的路要走。本文从解决CAR-T疗法毒副作用的关键技术——安全性控制出发,分析了该领域相关专利的技术发展路线,以期为我国创新主体在该领域的研发及产业化提供专利信息支持。

系统性红斑狼疮CAR T细胞治疗疗效预测及安全性评估的潜在生物标志物

系统性红斑狼疮(systemic lupus erythematosus,SLE)是一种复杂的自身免疫疾病,传统治疗在部分重度和难治性患者中效果有限。近期研究显示,嵌合抗原受体(chimeric antigen receptor,CAR)T细胞疗法在SLE治疗中展现出了具有前景的疗效。生物标志物在精准评估治疗效果和安全性方面至关重要,CAR T细胞治疗与安全性评估标志物包括传统标志物和与CAR T细胞疗法相关的标志物。传统的SLE病情监测标志物,仍可用于CAR T细胞治疗基线随访和病情监测,如血清抗双链DNA、抗单链DNA和抗核小体等自身抗体的滴度下降,血清补体水平恢复正常,以及尿蛋白/肌酐比值的改善,均提示病情得到有效控制。CAR T细胞疗效监测标志物分为B细胞和T细胞标志物。输注后,B细胞数量下降,B细胞表型以初始B细胞为主,记忆B细胞和浆母细胞的比例显著降低,表明治疗取得了疗效。输注前,初始T细胞(CD45RA+CD27+)和中央记忆型T细胞(CD45RA-CD62L+CD27+)的高比例则提示更强的抗肿瘤效应;患者的CAR T细胞表达与早期记忆分化相关的转录因子,如T细胞因子7和淋巴增强子结合因子1,提示这些患者对CAR T细胞疗法更为敏感。输注后,CD25、CD69和CD137等T细胞激活标志物,以及CD57、PD-1和Tim-3等耗竭标志物的高表达,提示T细胞的杀伤能力受到限制。CAR T细胞治疗安全性标志物不仅包括CAR T细胞分泌的效应细胞因子(如白细胞介素-2和IFN-γ),还包括单核细胞和巨噬细胞产生的细胞因子(如IL-1和IL-8),其水平可用于评估CAR T细胞疗法最常见的毒副反应[细胞因子释放综合征(cytokine release syndrome,CRS)和免疫效应细胞相关神经毒性综合征(immune effector cell-associated neurotoxicity syndrome,ICANS)]。高水平的血清巨噬细胞炎性蛋白1α对于预测CAR T细胞治疗后发生严重CRS和ICANS的风险具有较高的价值。此外,基线血小板计数和中性粒细胞绝对值可预测血液毒性,由IL-8、IFN-γ和IL-1β组成的感染相关预测模型,能够有效预测患者输注后出现严重感染的风险。CAR受体结构设计、清除淋巴细胞的化疗方式,患者曾接受的治疗选择及自身免疫状态等都会影响CAR T细胞治疗的疗效及安全性。在当前及未来将开启的相关临床研究中,应纳入全面、规范的检测和评估体系,为CAR T细胞疗法在SLE等自身免疫疾病的应用,提供比较标准。

City Routing Velocity Estimation Model under theEnvironment of Lack of Floating Car Data

After introducing the principle of float car data (FCD), this paper gives the primary flow of pre-handing and map- matching of the FCD. After analyzing the percentage of coverage of FCD on the road network, large quantity of heritage database of routing status is used to estimate the routing velocity when lack of FCD on parts road segments. Multi liner regression model is then put forwarded by considering the spatial correlativity among the road network, and some model parameters are deduced when time series is classified in day and week. Besides, error of velocity probability and error of status probability are achieved based on the result from field testing while the feasibility and reliability of the velocity estimation model is obtained as well. Finally, as a case study in Shanghai center area, the whole routing velocity in the road network is estimated and published in real time.

Phytoremediation Potential of Some Macrophytes from a Car Wash Stream in Buea, South Western Cameroon

Phytoremediation is a cheap and environmentally friendly technique in which green plants in situ are used to clean the soil, sediments and water of heavy metals. This study investigated the phytoremediation potential of six naturally occurring macrophytes from Nange (a stream in Buea municipality where cars have been driven into and washed for over 20 years). Plant samples were collected before and after car wash, then analysed for copper, zinc, lead and cadmium accumulation. There was an increase in concentration of all the four heavy metals in water after carwash point, with Zn having the highest concentration (0.27 mg/L). Mean concentration of the heavy metals in the water showed that Zn and Pb had the highest concentrations (0.24 mg/L each) while the least concentration was obtained in Cu (0.12 mg/kg). Heavy metal concentrations in the sediments were higher after car wash point than before. Cadmium had the highest concentration (5.58 mg/kg) while Cu had the least (0.75 mg/kg). Ludwigia peruviana had the highest BAFs for all the heavy metals (22.95 for Cu, 33.41 for Zn, 21.79 for Pb and 7.85 for Cd). Species with the leasts were: Anubias barteri for Cu (7.16), Polygonum persicaria for Zn (14.28), Rorippa nasturtium-aquaticum for Pb (11.60) and Vallisneria spiralis for Cd (1.98). L. peruviana had the highest BAC values (Cu = 10.11, Zn = 14.73, Pb = 11.39, Cd = 3.85) and BCF values (Cu = 12.84, Zn = 18.67, Pb = 10.40, Cd = 4.00). A. barteri had the highest TF (Cu = 1.49, Zn = 1.27, Cd = 1.99) except for Pb where both A. barteri and L. peruviana each had a TF of 1.10. While all the six plants were found to be good accumulators of the heavy metals, L. peruviana showed remarkable efficiency indicating that the species is a good candidate for cleaning such environments.

射血分数保留心衰后CAR-T细胞治疗的小鼠左心室应力应变分析

目的HFp EF已经成为心衰病例中的重要组成部分,其患病率逐年上升,且预后状况较差,对患者生活质量的影响显著。本文提出假设,CAR-T细胞可以有效治疗射血分数保留心衰心脏的心肌纤维化,并且通过分析治疗前后的左心室各区域应力应变来探索CAR-T细胞治疗的可能性。方法建立了心衰小鼠模型,分为假手术组(C)、心衰组(HF)、治疗组(T)。通过超声影像收集数据,通过几何重建与插值建立左室壁的三维几何模型,根据Fung给出的关于应变能的本构关系,对心室舒张进行模拟,最后通过有限元分析估算左室壁的应力和应变。结果心衰小鼠的左心室应力主要在内壁和基底面明显升高,而对照组和经过CAR-T细胞治疗后的左心室应力明显减少。心衰组的应变经治疗以后有明显增加。考虑到心肌细胞刚度等原因。CAR-T细胞治疗组的整体心肌刚度有明显降低,具有显著差异。结论结论是射血分数保留心衰可导致心脏室壁变厚、室腔扩大以及心室重构。射血分数保留心衰小鼠的左心室纤维化水平增高,左心纤维方向上的柯西应力增大,格林应变减小,心肌细胞刚度增加。经CAR-T细胞治疗后,小鼠左心纤维化明显减少,左心纤维方向上的应力减少,应变增大,心肌细胞刚度减小,恢复至正常对照组水平。

衰老细胞清除策略与CAR细胞疗法在抗衰老治疗中的应用

人类寿命逐渐延长,世界各国老年人口的数量和占比持续上升,但健康寿命却并未同步增长,衰老仍然是肿瘤、心血管疾病、阿尔茨海默病等疾病的主要风险因素。如何防治衰老相关疾病,延长老年人的健康寿命,仍亟待研究人员解决。细胞衰老是衰老的12大标志之一,存在于机体的整体衰老过程,可以说机体衰老的本质是组成机体的细胞的衰老,而通过转基因技术、基因重编程技术、使用新型抗衰老药物如senomorphic和senolytic等方法清除衰老细胞,已经成为目前延缓衰老、治疗衰老相关疾病、延长人类健康寿命的重要策略。免疫衰老是机体衰老的一部分,而免疫细胞是人体中最易、最先发生老化的细胞群体,针对免疫系统的抗衰老治疗例如抗体治疗、CAR细胞疗法、NK细胞疗法等是近年衰老研究的热点,研究认为,免疫疗法具有极高的临床抗衰老潜力。本文从细胞衰老和免疫衰老的特征入手,进而总结了目前衰老的非靶向干预方法和靶向衰老细胞清除的干预方法及其优缺点,并重点综述了靶向衰老细胞的免疫治疗清除策略。本文旨在阐明新兴衰老细胞清除方法的发展及局限,为防治衰老相关疾病、延长健康寿命提供新的临床应用策略。

有望刺激特殊基因来增强抵御人类多种实体瘤的CAR-T细胞疗法的疗效

据Chan JD 2024年4月10日(Nature,2024 Apr 10. doi:10.1038/s41586-024-07242-1.)报道,澳大利亚墨尔本大学等机构研究人员通过研究克服了一个主要障碍,利用一种新型基因工程策略,通过遗传方式赋予嵌合抗原受体(CAR)-T细胞有利的代谢特性,显著提升其在治疗人类实体瘤方面的效果。

冷冻干燥淋巴结载药系统可提升CAR-T细胞疗效

2024年3月6日,浙江大学药学院顾臻教授、李洪军研究员联合浙江大学医学院附属第一医院赵鹏主任医师团队在《自然·材料》(Nature Materials)上发表了最新研究成果“Lyophilized lymph nodes for improved delivery of chimeric antigen receptor T cells”(DOI:10.1038/s41563-024-01825-z)。该研究提出一种肌体组织工程化递药策略,使用冷冻干燥技术保留患者来源淋巴结的基质结构和关键免疫调节因子存储和递送CAR-T细胞,增强CAR-T细胞的杀伤活性和持久性。

Fatigue research of metro car body based on operational loads

Purpose–The principle of infinite life design currently directs fatigue resistance strategies for metro car bodies.However,this principle might not fully account for the dynamic influence of operational loads and the inevitable presence of defects.This study aims to integrate methods of service life estimation and residual life assessment,which are based on operational loads,into the existing infinite life verification framework to further ensure the operational safety of subway trains.Design/methodology/approach–Operational loads and fatigue loading spectra were determined through the field test.The material test was conducted to investigate characteristics of the fracture toughness and the crack growth rate.The fatigue strength of the metro car body was first verified using the finite element method and Moore–Kommers–Japer diagrams.The service life was then estimated by applying the Miner rule and high-cycle fatigue curves in a modified form of the Basquin equation.Finally,the residual life was assessed utilizing a fracture assessment diagram and a fitted curve of crack growth rate adhered to the Paris formula.Findings–Neither the maximum utilization factor nor the cumulative damage exceeds the threshold value of 1.0,the metro car body could meet the design life requirement of 30 years or 6.6 million km.However,three out of five fatigue key points were significantly influenced by the operational loads,which indicates that a single fatigue strength verification cannot achieve the infinite life design objective of the metro car body.For a projected design life of 30 years,the tolerance depth is 12.2 mm,which can underscore a relatively robust damage tolerance capability.Originality/value–The influence of operational loads on fatigue life was presented by the discrepancy analysis between fatigue strength verification results and service life estimation results.The fracture properties of butt-welded joints were tested and used for the damage tolerance assessment.The damage tolerance life can be effectively related by a newly developed equation in this study.It can be a valuable tool to provide the theoretical guidance and technical support for the structural improvements and maintenance decisions of the metro car body.

靶向EphA3的CAR-T细胞疗法有望治疗胶质瘤

据Lertsumitkul L[Journal for Immunotherapy of Cancer,2024,12(8):e009486-e009486.]报道,澳大利亚沃尔特和伊丽莎-霍尔医学研究所(WEHI)的研究人员发现了一种很有前景的二合一新疗法,它不仅能靶向摧毁一种侵袭性脑癌,还能帮助免疫系统形成对这种癌症的持久防御。这种双效疗法使用一种称为CAR-T细胞疗法的特殊免疫疗法来治疗胶质瘤,其中胶质瘤是一种难以治愈的脑癌,治疗方法很少。

将间质干细胞的线粒体转移给T细胞和CAR-T细胞可增强它们的抗癌功能

据Court AC (Journal of Translational Medicine,2024,22:868-868.)报道,智利洛斯安第斯大学IMPACT卓越中心的Maroun Khoury团队和巴西国家癌症研究所的Martín Bonamino团队公布了一种创新方法,通过为T细胞和CAR-T细胞提供线粒体超级充能来增强它们的功能。两项研究都强调了线粒体转移在提高T细胞性能方面的创新潜力。CAR-T细胞是一种经过改造的T细胞,可表达专门靶向和攻击癌细胞的受体。这种细胞疗法包括提取患者的T细胞,增强它们的功能,然后重新输注,以提高抗癌能力。研究人员发现,将间质干细胞(MSC)中的线粒体转移到T细胞中,可以显著提高T细胞的存活率和抗癌能力。

PC-CAR识别肿瘤靶标的结构原理

据Sun Y(Bio Rxiv,2023. doi:10.1101/2023.05.24.542108.)报道,美国费城儿童医院的研究人员解析出一种三维蛋白结构,解释了PC-CAR如何识别这些HLA复合物的“骨架”。这些结构信息将使得人们能够了解嵌合抗原受体(CAR)如何识别不同多态性HLA等位基因中的肿瘤相关抗原,从而为针对更复杂、更难治疗的肿瘤设计精准医疗策略提供更多可能性。

Estimation of speed-related car body acceleration limits with quantile regression

Purpose–This study aimed to facilitate a rapid evaluation of track service status and vehicle ride comfort based on car body acceleration.Consequently,a low-cost,data-driven approach was proposed for analyzing speed-related acceleration limits in metro systems.Design/methodology/approach–A portable sensing terminal was developed to realize easy and efficient detection of car body acceleration.Further,field measurements were performed on a 51.95-km metro line.Data from 272 metro sections were tested as a case study,and a quantile regression method was proposed to fit the control limits of the car body acceleration at different speeds using the measured data.Findings–First,the frequency statistics of the measured data in the speed-acceleration dimension indicated that the car body acceleration was primarily concentrated within the constant speed stage,particularly at speeds of 15.4,18.3,and 20.9 m/s.Second,resampling was performed according to the probability density distribution of car body acceleration for different speed domains to achieve data balance.Finally,combined with the traditional linear relationship between speed and acceleration,the statistical relationships between the speed and car body acceleration under different quantiles were determined.We concluded the lateral/vertical quantiles of 0.8989/0.9895,0.9942/0.997,and 0.9998/0.993 as being excellent,good,and qualified control limits,respectively,for the lateral and vertical acceleration of the car body.In addition,regression lines for the speedrelated acceleration limits at other quantiles(0.5,0.75,2s,and 3s)were obtained.Originality/value–The proposed method is expected to serve as a reference for further studies on speedrelated acceleration limits in rail transit systems.