Using Drosophila melanogaster to Analyse the Human Paralogs of the ESCRT-III Core Component Shrub/CHMP4/Snf7 and Its Interactions with Members of the LGD/CC2D1 Family

The evolutionary conserved ESCRT-III complex is a device for membrane remodelling in various cellular processes, such as the formation of intraluminal vesicles (ILVs), cytokinesis, and membrane repair. The common theme of all these processes is the abscission of membrane away from the cytosol. At its heart in Drosophila is Shrub, CHMP4 in humans, which dynamically polymerises into filaments through electrostatic interactions among the protomers. For the full activity, Shrub/CHMP4 requires physical interaction with members of the Lgd protein family. This interaction is mediated by the odd-numbered DM14 domains of Lgd, which bind to the negative interaction surface of Shrub. While only one Lgd and one Shrub exist in the genome of Drosophila, mammals have two Lgd orthologs, LGD1/CC2D1B and LGD2/CC2D1A, as well as three CHMP4s in their genomes, CHMP4A, CHMP4B, and CHMP4C. The rationale for the diversification of the ESCRT components is not understood. We here use Drosophila as a model system to analyse the activity of the human orthologs of Shrub and Lgd at an organismal level. This enabled us to use the plethora of available techniques available for Drosophila. We present evidence that CHMP4B is the true ortholog of Shrub, while CHMP4A and CHMP4C have diverging activities. Nevertheless, CHMP4A and CHMP4C can enhance the activity of CHMP4B, raising the possibility that they can form heteropolymers in vivo. Our structure-function analysis of the LGD1 and LGD2 indicates that the C2 domain of the LGD proteins has a specific function beyond protein stability and subcellular localisation. Moreover, our data specify that CHMP4B interacts more efficiently with LGD1 than with LGD2.     

Clinically Applicable Assessment of Tisagenlecleucel CAR T Cell Treatment by Digital Droplet PCR for Copy Number Variant Assessment

Chimeric antigen receptor (CAR) T cell therapy is an innovative immunotherapy for treating cancers in both children and adults with proven utility in numerous clinical trials. Significantly, some CAR T cell therapies have now been approved by relevant national regulatory bodies across numerous countries for clinical therapeutic use outside of clinical trials. One such recently licensed product is tisagenlecleucel, a CAR T therapy approved for the treatment of B-cell acute lymphoblastic leukemia (B-ALL) using autologous T cells from the patient. The genetically engineered T cells target a protein called CD19, common to B cells, through a CAR incorporating a 4-1BB costimulatory domain to improve response. Since tisagenlecleucel is now a standard of care treatment for B-ALL, it is clinically essential to be able to accurately monitor these CAR T cells in patients. Assessment of the copy number variant (CNV) of the CAR T cell products allows this within a clinically acceptable timeframe for optimal patient benefit. However, no standardized method with high reproducibility and efficiency has been described within a routine clinical laboratory setting. Here, we demonstrated a novel digital droplet PCR (ddPCR)-based methodology for the study of CNV (ddPCR-CNV) in 4-1BB CD19-specific CAR T cells with universal applicability across clinical diagnostic laboratories.     

水溶液中4-(2-羟基-苯甲酸酯基)-2-环戊烯砜的合成

以3,4-二溴环戊砜(1)为原料,在无水吡啶作用下发生消除反应,得到反应中间体4-溴-2-环戊烯砜(2),再与水杨酸钠(3)在水溶液和DMF中发生酯化反应,合成水杨酸酯环戊烯砜衍生物(4),用IR,1H NMR,MS,元素分析表征了它们的结构,并讨论了酯化反应在在水溶液和DMF中的最佳反应条件。结果表明:水溶液中的酯化反应后处理更加简单,产率最高。     

Impact of High-Molecular-Weight Hyaluronic Acid on Gene Expression in Rabbit Achilles Tenocytes In Vitro

(1) Background: Surgical tendon repair often leads to adhesion formation, leading to joint stiffness and a reduced range of motion. Tubular implants set around sutured tendons might help to reduce peritendinous adhesions. The lubricant hyaluronic acid (HA) is a viable option for optimizing such tubes with the goal of further enhancing the anti-adhesive effect. As the implant degrades over time and diffusion is presumed, the impact of HA on tendon cells is important to know. (2) Methods: A culture medium of rabbit Achilles tenocytes was supplemented with high-molecular-weight (HMW) HA and the growth curves of the cells were assessed. Additionally, after 3, 7 and 14 days, the gene expression of several markers was analyzed for matrix assembly, tendon differentiation, fibrosis, proliferation, matrix remodeling, pro-inflammation and resolution. (3) Results: The addition of HA decreased matrix marker genes, downregulated the fibrosis marker α-SMA for a short time and slightly increased the matrix-remodeling gene MMP-2. Of the pro-inflammatory marker genes, only IL-6 was significantly upregulated. IL-6 has to be kept in check, although IL-6 is also needed for a proper initial inflammation and efficient resolution. (4) Conclusions: The observed effects in vitro support the intended anti-adhesion effect and therefore, the use of HMW HA is promising as a biodegradable implant for tendon repair.     

4D 打印技术在舰炮武器装备维修保障中的应用

为适应未来信息化作战的需要,优化舰炮武器装备的维修保障工作,探讨4D 打印技术在全寿命周期维 修保障中的应用。将舰炮武器装备的维修保障提前至论证决策阶段,并贯穿全寿命周期,分析4D 打印技术原理, 梳理维修保障的关键问题,以期拓展信息化作战条件下舰炮武器装备的维修保障工作思路。结果表明,4D 打印技术 为相关研制部门和一线作战使用单位提供参考依据。     

基于窄带物联网的水务管网参数监测系统设计

近几年随着城市供水管道的日益复杂化,城市供水系统的监测变得难以管理.为了更好地监测城市供水系统的运行状态,借助窄带物联网(Narrow Band Internet of Things,NB-IoT)的低功耗优势,设计了一个基于NB-IoT的水务管网参数监测系统.该系统以低功耗STM32L4芯片为控制核心,实现了供水管网...     

含二苯并-14-冠-4冠醚环主链型液晶共聚酯的设计与合成

以4,4-′(α,ω-亚烷基二酰氧)二联苯甲酰氯(M1)、顺(反)式-4,4′-双(4-羟基苯基偶氮)二苯并-14-冠-4(M2、M3)和1,10-癸二醇(M4)为单体,通过溶液共缩聚反应,合成了两个系列新的含冠醚环的主链型液晶共聚酯。共聚酯的分子量不高。单体的化学结构通过IR、UV-Vis1、H-NMR和元素分析等方法确证。共聚酯的性质采用[η]、DSC、TG、WAXD和POM等方法进行了研究。发现所有的共聚酯加热到各自的熔融温度以上都能形成液晶态,在液晶态可以观察到向列相的丝状织构或纹影织构。共聚酯的熔融温度(Tm)和各向同性温度(Ti)随共聚酯分子中柔性间隔基的改变呈规律性变化。     

低带隙共轭聚合物的合成及光学性质

在钯催化剂的作用下,通过4,7-二溴[2,1,3]苯并噻二唑与2,5-二乙炔基-3-辛基噻吩的偶联反应,合成了一种新的共轭高分子PTE-BT.采用紫外可见吸收光谱及荧光光谱对其光学性质进行了研究.紫外可见吸收谱结果表明,PTE-BT的固体膜光学带隙为1.86 ev;TiO2/PTE-BT共混固体膜的荧光发射谱结果表明,电子供体PTE-BT分子与电子受体TiO2分子间存在有效的电子转移.     

Gd2O2SO4粉体的合成及其晶体结构确定

通过750℃,2h煅烧Gd2O3和H2SO4(摩尔比1:1)的前驱体方法合成了Gd2O2SO4粉体.利用Gd2O2SO4的X射线粉末衍射数据和Material Studio 4.0 Reflex模块中Powder solve技术和Rietveld精修方法对Gd2O2SO4的晶体结构进行了研究.研究表明:Gd2O2SO4具有空间群为PMNB(62)的正交晶系,晶胞参数为a=1.2996nm,b=0.8117nm,c=0.4184nm,V=0.440698nm3,Z=4,计算密度为6.6949g/cm3,并确定了晶胞中原子的位置,Rietveld精修得到的Rwp=9.15%.