Interdependence of Angiogenesis and Arteriogenesis in Development and Disease

The structure of arterial networks is optimized to allow efficient flow delivery to metabolically active tissues. Optimization of flow delivery is a continuous process involving synchronization of the structure and function of the microcirculation with the upstream arterial network. Risk factors for ischemic cardiovascular diseases, such as diabetes mellitus and hyperlipidemia, adversely affect endothelial function, induce capillary regression, and disrupt the micro- to macrocirculation cross-talk. We provide evidence showing that this loss of synchronization reduces arterial collateral network recruitment upon arterial stenosis, and the long-term clinical outcome of current revascularization strategies in these patient cohorts. We describe mechanisms and signals contributing to synchronized growth of micro- and macrocirculation in development and upon ischemic challenges in the adult organism and identify potential therapeutic targets. We conclude that a long-term successful revascularization strategy should aim at both removing obstructions in the proximal part of the arterial tree and restoring “bottom-up” vascular communication.     

Low Temperature Decoherence by Electron–Electron Interactions: Role of Quantum Fluctuations

We derive a general expression for the conductivity of a disordered conductor with electron–electron interactions (treated within the standard model) and evaluate the weak localization correction _wl employing no approximations beyond the accuracy of the definition of _wl . Our analysis applies to all orders in the interaction and extends our previous calculation by explicitly taking into account quantum fluctuations around the classical paths for interacting electrons (pre-exponent). We specifically address the most interesting low temperature limit and demonstrate that such fluctuations can only be important in the perturbative regime of short times while they are practically irrelevant for the Cooperon dynamics at longer times. We fully confirm our conclusion about the existence of interaction-induced decoherence of electrons at zero temperature for the problem in question. We also demonstrate irrelevance of a perturbative calculation by Aleiner et al. (AAV) [J. Low Temp. Phys. 126, 1377 (2002)] and refute AAV's critique of our earlier analysis.     

重组2型腺相关病毒肿瘤坏死因子相关凋亡诱导配体基因治疗制剂的质量分析

目的对重组2型腺相关病毒肿瘤坏死因子相关凋亡诱导配体(recombinant adeno-associated virus 2 encoding tumor necrosis factor-related apoptosis-induced ligand,r AAV2-TRAIL)基因治疗制剂进行质量分析,并针对其关键质量属性建立质控方法。方法采用PCR法对r AAV2-TRAIL基因治疗制剂中插入的启动子CAG和目的基因TRAIL进行鉴别;在腺病毒(Ad5)的辅助下,将r AAV2-TRAIL体外感染293T细胞后,采用ELISA法检测培养上清中目的蛋白TRAIL的表达量,检测r AAV2-TRAIL对神经胶质瘤U251细胞的体外杀伤活性;设计引物及探针,采用Q-PCR法检测r AAV2-TRAIL基因治疗制剂中的r AAV2-TRAIL滴度、可能存在的复制型rc AAV滴度以及残余辅助病毒1型单纯疱疹病毒(herpes simplex virus 1,HSV1)滴度。结果启动子CAG和目的基因TRAIL的PCR鉴定结果与理论及理化对照品相符;r AAV2-TRAIL感染293T细胞48 h后,上清中TRAIL表达量为(0.36±0.18)ng/ml,RSD为50.0%;r AAV2-TRAIL体外作用于神经胶质瘤细胞U251,细胞生长相对抑制率为(26.6±3.75)%,RSD为14.1%;制剂中r AAV2-TRAIL滴度为(4.72×1011±0.52×1011)copies/ml,RSD为11.0%,rc AAV滴度为(2.49×107±0.18×107)copies/ml,RSD为7.2%,HSV1滴度为(6.02×106±0.51×106)copies/ml,RSD为8.5%。r AAV2-TRAIL/rc AAV为1.90×104,r AAV2-TRAIL/HSV1为7.84×104。结论建立的质控方法可用于r AAV2-TRAIL的质量控制,为该产品质量标准的建立奠定了基础,同时对以AAV为载体的基因治疗制剂的质量研究提供参考。     

抗中性粒细胞胞质抗体相关性小血管炎的机制与治疗进展

血管炎是一类可引起血管非特异性炎症和坏死的自身免疫性疾病。按血管受累的大小分类,包括了小血管炎、中等血管炎及大血管炎。抗中性粒细胞胞质抗体（ANCA）相关性小血管炎（AAV）属于小血管炎的范畴,主要包含了肉芽肿性多血管炎（GPA）,显微镜下多血管炎（MPA）和嗜酸肉芽肿性多血管炎（EGPA）。它们可累及全身不同脏器,活动期异常凶险,治疗不及时可致高死亡率。本文简要综述AAV的发病机制及治疗进展。     

整合siRNA技术的结核核酸疫苗的AAV载体构建与浓缩纯化

引进腺病毒相关病毒载体系统(AAV载体系统),并通过PCR、酶切、连接等分子克隆手段将之前对于结核研究的成果转移至该系统中,完成同时具备小干扰RNA( si-Mcl-1)和结核融合抗原(AG85B-ESAT6)表达单位的二重表达载体的构建.转染AAV-293细胞,包装重组AAV病毒颗粒(rAAV)并收集病毒.分别设计另...     

新型塔内件在TDI第一脱ODCB塔扩产改造中的应用

针对蓝星化工有限责任公司TDI精制装置中第一脱邻二氯苯（ODCB）塔在扩产改造中存在的设计问题,在工艺模拟和流体力学计算的基础上,选择新型的塔内件和填料对上述设备进行再次改造。将原规整填料更换为DZ波纹高效规整填料,液体分布器设计为带垂直布液板的二级悬槽式液体分布器,回流温度由40℃提高至60～65℃,在不增加塔高的情况下,第一脱ODCB塔理论塔板数由6块增加到20块,回收的ODCB溶剂中平均胺吸收值（AAV）小于0．02％,ODCB的纯度由95％提高99．98％以上,TDI产能由2万t／a提高到3万t／a,取得了良好的经济和社会效益。     

Therapeutic Applications of Adeno-Associated Virus (AAV) Gene Transfer of HLA-G in the Eye

The purpose of this paper is to review human leukocyte antigen G (HLA-G) in the eye, its role in immune tolerance, and the potential therapeutic use of AAV gene transfer and expression of HLA-G in various ocular tissues. Several studies are reviewed that demonstrate efficacy in animal models of disease, including intracorneal delivery of AAV-HLA-G to treat corneal inflammation and prevent corneal graft rejection, subconjunctival injection of AAV-HLA-G for ocular graft vs. host disease and potentially dry eye disease, and intravitreal injection of AAV-HLA-G to inhibit uveitis. Furthermore, due to the anti-vascular function of HLA-G, AAV-HLA-G may be an effective therapy for posterior ocular diseases, such as neovascular age-related macular degeneration, diabetic retinopathy, and choroidal neovascularization. Therefore, AAV-mediated gene transfer of HLA-G may be an effective treatment for common immune-mediated, inflammatory, and neovascular diseases of the eye.     

空气热源式气化技术在大型LNG接收终端的应用

近年来,空气热源式气化设备和技术（Ambient air based heating vaporization,AHV）因其具有环境友好、节约能源、可持续利用等优势而在其他国家的新建LNG项目中不断得到实践验证和推行。为此,介绍了AHV的技术特点、分类及其在世界各地的最新应用情况,并与常规气化技术进行了对比,进而提出了该类技术的应用条件,分析了该技术方案的优缺点。结果认为：①LNG接收终端所处位置具体环境的气象条件（年最低环境温度、湿度、风向和风速、持续时间等）是选择合适的AHV技术的关键因素;②设计空气热源气化系统需要确定它的最低环境温度以及备用热源系统需要热机备用的工况条件;③相对于浸没燃烧式气化器,AHV系统的优势明显,且燃气（电）价格比率越高,其优势越显著;④在不适合采用海水开架式气化技术的条件下,AHV可作为优选方案;⑤在免费海水使用受限、天然气燃烧制热成本高昂的情况下,以空气热源作为气化系统基荷热源是最为便利和直接的选择。为中国规划和设计新建LNG接收终端提供了更为经济和环保的气化技术选项     

Sustained Elevated Blood Pressure Accelerates Atherosclerosis Development in a Preclinical Model of Disease

The continuous relationship between blood pressure (BP) and cardiovascular events makes the distinction between elevated BP and hypertension based on arbitrary cut-off values for BP. Even mild BP elevations manifesting as high-normal BP have been associated with cardiovascular risk. We hypothesize that persistent elevated BP increases atherosclerotic plaque development. To evaluate this causal link, we developed a new mouse model of elevated BP based on adeno-associated virus (AAV) gene transfer. We constructed AAV vectors to support transfer of the hRenin and hAngiotensinogen genes. A single injection of AAV-Ren/Ang (10¹¹ total viral particles) induced sustained systolic BP increase (130 ± 20 mmHg, vs. 110 ± 15 mmHg in controls; p = 0.05). In ApoE^−/− mice, AAV-induced mild BP elevation caused larger atherosclerotic lesions evaluated by histology (10-fold increase vs. normotensive controls). In this preclinical model, atheroma plaques development was attenuated by BP control with a calcium channel blocker, indicating that a small increase in BP within a physiological range has a substantial impact on plaque development in a preclinical model of atherosclerosis. These data support that non-optimal BP represents a risk for atherosclerosis development. Earlier intervention in elevated BP may prevent or delay morbidity and mortality associated with atherosclerosis.     

Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors

A major limiting factor for systemically delivered gene therapies is the lack of novel tissue specific AAV (Adeno-associated virus) derived vectors. Bispecific antibodies can be used to redirect AAVs to specific target receptors. Here, we demonstrate that the insertion of a short linear epitope “2E3” derived from human proprotein-convertase subtilisin/kexin type 9 (PCSK9) into different surface loops of the VP capsid proteins can be used for AAV de-targeting from its natural receptor(s), combined with a bispecific antibody-mediated retargeting. We chose to target a set of distinct disease relevant membrane proteins—fibroblast activation protein (FAP), which is upregulated on activated fibroblasts within the tumor stroma and in fibrotic tissues, as well as programmed death-ligand 1 (PD-L1), which is strongly upregulated in many cancers. Upon incubation with a bispecific antibody recognizing the 2E3 epitope and FAP or PD-L1, the bispecific antibody/rAAV complex was able to selectively transduce receptor positive cells. In summary, we developed a novel, rationally designed vector retargeting platform that can target AAVs to a new set of cellular receptors in a modular fashion. This versatile platform may serve as a valuable tool to investigate the role of disease relevant cell types and basis for novel gene therapy approaches.