The roles of intrinsic disorder-based liquid-liquid phase transitions in the “Dr. Jekyll–Mr. Hyde” behavior of proteins involved in amyotrophic lateral sclerosis and frontotemporal lobar degeneration

Pathological developments leading to amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are associated with misbehavior of several key proteins, such as SOD1 (superoxide dismutase 1), TARDBP/TDP-43, FUS, C9orf72, and dipeptide repeat proteins generated as a result of the translation of the intronic hexanucleotide expansions in the C9orf72 gene, PFN1 (profilin 1), GLE1 (GLE1, RNA export mediator), PURA (purine rich element binding protein A), FLCN (folliculin), RBM45 (RNA binding motif protein 45), SS18L1/CREST, HNRNPA1 (heterogeneous nuclear ribonucleoprotein A1), HNRNPA2B1 (heterogeneous nuclear ribonucleoprotein A2/B1), ATXN2 (ataxin 2), MAPT (microtubule associated protein tau), and TIA1 (TIA1 cytotoxic granule associated RNA binding protein). Although these proteins are structurally and functionally different and have rather different pathological functions, they all possess some levels of intrinsic disorder and are either directly engaged in or are at least related to the physiological liquid-liquid phase transitions (LLPTs) leading to the formation of various proteinaceous membrane-less organelles (PMLOs), both normal and pathological. This review describes the normal and pathological functions of these ALS- and FTLD-related proteins, describes their major structural properties, glances at their intrinsic disorder status, and analyzes the involvement of these proteins in the formation of normal and pathological PMLOs, with the ultimate goal of better understanding the roles of LLPTs and intrinsic disorder in the “Dr. Jekyll–Mr. Hyde” behavior of those proteins.     

A novel bispecific molecule delivered by recombinant AAV2 suppresses ocular inflammation and choroidal neovascularization

Elevated vascular endothelial growth factor (VEGF) and complement activation are implicated in the pathogenesis of different ocular diseases. The objective of this study was to investigate the hypothesis that dual inhibition of both VEGF and complement activation would confer better protection against ocular inflammation and neovascularization. In this study, we engineered a secreted chimeric VEGF inhibitor domain (VID), a complement inhibitor domain (CID) and a dual inhibitor (ACVP1). Vectors expressing these three inhibitors were constructed and packaged into AAV2 (sextY‐F) particles. The expression and secretion of the proteins were validated by Western blot. The effects of these inhibitors expressed from AAV2 vectors were examined in endotoxin‐induced uveitis (EIU), experimental autoimmune uveoretinitis (EAU) and choroidal neovascularization (CNV) mouse models. The AAV2 vectors expressing the CID‐ and ACVP1‐attenuated inflammation in EIU and EAU model, whereas the vector expressing VID showed improved retinal structure damaged by EAU, but not affect the infiltration of inflammatory cells in EAU or EIU eyes. Both VID and CID vectors improved laser‐induced retinal and choroid/RPE injuries and CNV, whereas ACVP1 vector provided significantly better protection. Our results suggest that gene therapy targeting VEGF and complement components could provide an innovative and long‐term strategy for ocular inflammatory and neovascular diseases.     

腰椎旁神经阻滞联合超短波对腰椎间盘突出症患者疼痛及腰背肌生物力学性能的影响

目的:探讨腰椎旁神经阻滞联合超短波对腰椎间盘突出症疼痛及腰背肌生物力学性能的影响。方法:选择我院2014年2月~2016年8月收治的98例腰椎间盘突出症患者,按抽签法分组对照组与研究组。对照组采用腰椎旁神经阻滞治疗,研究组基于对照组加用超短波治疗。观察两组的临床疗效、治疗前后视觉模拟评分(VAS)、60°/s角速、120°/s角速平均功率(AP)、峰力矩(PT)、腰背屈/伸比值(F/E)、血清P物质(SP)、β-内啡肽(β-EP)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平的变化及不良反应的发生情况。结果:研究组总有效率为95.91%,显著高于对照组,差异有统计学意义(P0.05)。治疗后,两组F/E值、血清SP、IL-6、TNF-α水平均较治疗前显著降低,且研究组以上指标均明显低于对照组,两组AP、PT、血清β-EP水平均较治疗前明显上升,且研究组以上指标显著高于对照组,差异均有统计学意义(P0.05)。两组不良反应的发生率比较差异无统计学意义(P0.05)。结论:腰椎旁神经阻滞联合超短波治疗腰椎间盘突出症的效果明显优于单用腰椎旁神经阻滞治疗,其可有效缓解疼痛及改善腰背肌生物力学性能,并减轻炎症反应。     

多节段腰椎间盘突出症的责任节段治疗与整体治疗的临床疗效比较

目的:研究和比较不同的治疗方式(责任节段治疗与整体治疗)治疗多节段腰椎间盘突出症(Herniation of Mutisegmented Lumber Intervertebral Disc,HMLD)的临床疗效及安全性。方法:对2010.01年至2013.01在我科明确诊断为多节段腰椎间盘突出且行手术治疗的共计78例患者进行回顾性分析。按照治疗方式的不同分为责任节段组(实验组,42例)和整体治疗组(对照组,36例)。结合手术前后的随访资料,评价并比较两组患者的疼痛模拟评分(VAS)、JOA功能评分及围手术期的手术时间、术中出血量、术后下地时间,花费,并发症等指标。结果:实验结果显示,实验组与对照组在术后6月,12月及36月的VAS疼痛评分及JOA评分的比较中并无显著性差异(P0.05)。但术后第二日实验组患者的疼痛程度显著好于对照组(P0.05)。实验组的术中出血量、花费及下地时间显著优于对照组(P0.05)。在并发症方面,术后1年内,实验组的并发症发生率显著优于对照组(P0.05);术后1年后,对照组的发生率较好,但两组间均无统计学差异(P0.05)。结论:对于多节段椎间盘突出症,找到责任节段并针对责任节段进行治疗较整体治疗来讲,能够在取得相似治疗效果和安全性的同时,能够有效的减少花费,手术创伤及术后疼痛指标,并能有效减少短期并发症的发生。在多节段腰椎间盘突出症的治疗中可以作为一种推荐的手术术式。     

Protein engineering to increase the potential of a therapeutic antibody Fab for long-acting delivery to the eye

To date, ocular antibody therapies for the treatment of retinal diseases rely on injection of the drug into the vitreous chamber of the eye. Given the burden for patients undergoing this procedure, less frequent dosing through the use of long-acting delivery (LAD) technologies is highly desirable. These technologies usually require a highly concentrated formulation and the antibody must be stable against extended exposure to physiological conditions. Here we have increased the potential of a therapeutic antibody antigen-binding fragment (Fab) for LAD by using protein engineering to enhance the chemical and physical stability of the molecule. Structure-guided amino acid substitutions in a negatively charged complementarity determining region (CDR-L1) of an anti-factor D (AFD) Fab resulted in increased chemical stability and solubility. A variant of AFD (AFD.v8), which combines light chain substitutions (VL-D28S:D30E:D31S) with a substitution (VH-D61E) to stabilize a heavy chain isomerization site, retained complement factor D binding and inhibition potency and has properties suitable for LAD. This variant was amenable to high protein concentration (>250 mg/mL), low ionic strength formulation suitable for intravitreal injection. AFD.v8 had acceptable pharmacokinetic (PK) properties upon intravitreal injection in rabbits, and improved stability under both formulation and physiological conditions. Simulations of expected human PK behavior indicated greater exposure with a 25-mg dose enabled by the increased solubility of AFD.v8.     

人类Y染色体的演化

人类Y染色体由于其性别决定的特殊功能和独有的进化史一直以来都备受关注。Y染色体起源于常染色体,经历了严重的退化过程。由于其缺乏重组,蛋白编码基因少,重复序列多所以研究进展缓慢。近年来,随着比较基因组及测序技术的快速发展,对人类Y染色体最终命运的争论不断加剧,Y染色体的研究正逐步成为热点。文章综述了人类Y染色体的结构、遗传特点、起源及进化过程,并根据目前的研究进展对Y染色体的最终命运进行了讨论,提出了作者的一些看法,以期为从事遗传及性染色体进化的研究者提供参考。     

果蝇翅原基发育分化的主要过程及分子机理

翅原基发育分化与昆虫的个体发育紧密联系, 对昆虫翅发育的研究有助于阐述昆虫的发育过程。另外, 翅的形成是一些农林害虫泛滥的主要原因之一, 研究翅发育分化有助于我们从翅发育的角度控制农林害虫。目前, 翅发育分化在果蝇Drosophila中研究已较为深入详细。果蝇翅发育分化主要包括4个阶段： 翅原基（wing disc）的确定, 前-后（antero-posterior, A-P）和背-腹（dorso-ventral, D-V）组织中心（organizing center）的建立, 翅区（wing region）的确定, 以及翅区的进一步分化。具有homeobox序列的基因（homeobox 基因）如Engrailed (En)、 Apterous (Ap)和Ultrabithorax (Ubx), 分泌蛋白如Wnt家族成员Wingless (Wg)及TGF-β超家族成员Decapentaplegic (Dpp)和Hedgehog (Hh), 以及翅原基特有的核蛋白编码基因Vestigial (Vg), 共同调控了翅原基的正常发育分化。本文综述了果蝇翅原基发育分化的过程及分子机理方面的研究发现, 为翅原基的研究提供了参考。     

Modeling The Nutrientsbehavior in Intervertebral Discs: A Boundary Integral Simulation

It is a well-known fact that computational biomechanics and mechanobiology have deserved great attention by the numerical-methods community. Many efforts and works can be found in technical literature. This work deals with the modeling of nutrients and their effects on the behavior of intervertebral discs. The numerical modeling was carried out using the Boundary ELement Method (BEM) and an axisymmetric model of the disc. Concentration and production of lactate and oxygen are modeled with the BEM. Results agree well enough with those obtained using finite elements. The numerical efforts in the domain and boundary discretizations are minimized using the BEM. Also, the effect of the calcification of the disc that causes the vascularization loss has been studied. The glucose, oxygen and lactate components behavior has been analyzed applying a mixed loading-unloading process, then allowing the study of the disc-height variations due to the degradation of the disc.     

冬虫夏草菌丝提取物对化学性肝损伤的辅助保护作用

为明确冬虫夏草菌丝提取物对急性肝损伤小鼠谷丙转氨酶（ALT）、谷草转氨酶（AST）、肝细胞变性及坏死程度的影响,采用四氯化碳（CCl4）诱导小鼠急性化学性肝损伤模型,将动物随机分成5组,分别是空白对照组、模型组、冬虫夏草菌丝提取物低剂量组（1.11g/kg BW）、中剂量组（3.33g/kg BW）、高剂量组（10.00g/kg BW）,检测血清ALT、AST值,并取肝脏作病理切片,观察肝脏的病理损伤情况。冬虫夏草菌丝提取物高剂量组能明显降低CCl4急性肝损伤小鼠血清ALT值,减轻肝细胞坏死程度,表明冬虫夏草菌丝提取物对化学性肝损伤有辅助保护功能。