Biochemical and structural characterization of hepatitis A virus 2C reveals an unusual ribonuclease activity on single-stranded RNA

The HAV nonstructural protein 2C is essential for virus replication; however, its precise function remains elusive. Although HAV 2C shares 24–27% sequence identity with other 2Cs, key motifs are conserved. Here, we demonstrate that HAV 2C is an ATPase but lacking helicase activity. We identified an ATPase-independent nuclease activity of HAV 2C with a preference for polyuridylic single-stranded RNAs. We determined the crystal structure of an HAV 2C fragment to 2.2 Å resolution, containing an ATPase domain, a region equivalent to enterovirus 2C zinc-finger (ZFER) and a C-terminal amphipathic helix (PBD). The PBD of HAV 2C occupies a hydrophobic pocket (Pocket) in the adjacent 2C, and we show the PBD–Pocket interaction is vital for 2C functions. We identified acidic residues that are essential for the ribonuclease activity and demonstrated mutations at these sites abrogate virus replication. We built a hexameric-ring model of HAV 2C, revealing the ribonuclease-essential residues clustering around the central pore of the ring, whereas the ATPase active sites line up at the gaps between adjacent 2Cs. Finally, we show the ribonuclease activity is shared by other picornavirus 2Cs. Our findings identified a previously unfound activity of picornavirus 2C, providing novel insights into the mechanisms of virus replication.     

An insight into planarian regeneration

BackgroundPlanarian has attracted increasing attentions in the regeneration field for its usefulness as an important biological model organism attributing to its strong regeneration ability. Both the complexity of multiple regulatory networks and their coordinate functions contribute to the maintenance of normal cellular homeostasis and the process of regeneration in planarian. The polarity, size, location and number of regeneration tissues are regulated by diverse mechanisms. In this review we summarize the recent advances about the importance genetic and molecular mechanisms for regeneration control on various tissues in planarian.MethodsA comprehensive literature search of original articles published in recent years was performed in regards to the molecular mechanism of each cell types during the planarian regeneration, including neoblast, nerve system, eye spot, excretory system and epidermal.ResultsAvailable molecular mechanisms gave us an overview of regeneration process in every tissue. The sense of injuries and initiation of regeneration is regulated by diverse genes like follistatin and ERK signaling. The Neoblasts differentiate into tissue progenitors under the regulation of genes such as egfr‐3. The regeneration polarity is controlled by Wnt pathway, BMP pathway and bioelectric signals. The neoblast within the blastema differentiate into desired cell types and regenerate the missing tissues. Those tissue specific genes regulate the tissue progenitor cells to differentiate into desired cell types to complete the regeneration process.ConclusionAll tissue types in planarian participate in the regeneration process regulated by distinct molecular factors and cellular signaling pathways. The neoblasts play vital roles in tissue regeneration and morphology maintenance. These studies provide new insights into the molecular mechanisms for regulating planarian regeneration.

Genetic and molecular mechanisms for regeneration control on various tissues in planarian.     

温度诱导Targetron系统用于大肠杆菌高效基因失活

构建基于Te I3c/4c嗜热二型内含子的温度诱导Targetron基因失活系统(Thermotargetron),并应用于中温微生物基因编辑。在大肠杆菌HMS174(DE3)基因组中,选择Subunitofflagellum基因(fliC)和C4dicarboxylate orotate:H+symporter基因(dctA)为靶基因。根据Te I3c/4c DNA识别规则,在fliC和dctA基因中选择fliC489a、fliC828s、fliC1038s和dctA2a位点为基因打靶位点。使用重叠延伸PCR方法,基于pHK-TT1A质粒构建打靶载体。打靶载体转化HMS174菌株,对数期转化子培养液48℃热激1h后涂布于氯霉素抗性LB平板上。使用菌落PCR和DNA测序检测突变株并计算基因失活效率。获得突变株后,通过琼脂穿刺和碳源代谢实验,鉴定ΔfliC、ΔdctA突变株表型变化。菌落PCR测序结果表明,Te I3c/4c插入到fliC和dctA基因设计位点,且打靶效率高达100%。突变株表型验证实验表明,ΔfliC突变株运动能力显著下降,ΔdctA突变株苹果酸代谢能力缺失。综上所述,...     

细胞重编程和移植技术用于帕金森病治疗的研究进展

帕金森病(Parkinson disease,PD)是一种复杂的中枢神经系统退行性疾病,主要病理特征为黑质致密部多巴胺神经元的进行性丧失.目前PD主要治疗手段包括药物和手术.但药物存在神经保护活性不足、缺乏对因治疗、晚期无药可用等问题,手术治疗风险较大.近年来,细胞重编程技术取得突破性进展,由重编程产生的诱导多能干细胞(induced pluripotent stem cells,iPSCs)、诱导多巴胺神经元(induced dopamine neurons,iDNs)和诱导神经干细胞(induced neural stem cells,i NSCs)可用于治疗PD.移植iPSCs分化而来的多巴胺能神经元、iDNs和iNSCs至相应脑区,可起到神经替代与修复作用,有效治疗PD.本文重点介绍细胞重编程的机制,总结iPSCs、iDNs和iNSCs治疗PD的优缺点,并阐述尚存在的挑战,探讨可能的解决方案.     

Ginsenoside Rh2 mitigates doxorubicin‐induced cardiotoxicity by inhibiting apoptotic and inflammatory damage and weakening pathological remodelling in breast cancer‐bearing mice

ObjectivesThere are presently a few viable ways to reduce cardiotoxicity of doxorubicin (Dox). The combination of chemotherapy agents with natural compounds delivers greater efficacy and reduces adverse effects in recent researches for cancer treatment. Here, we examined the potential effect of ginsenoside Rh2 on a Dox‐based regimen in chemotherapy treatment.Materials and MethodsHuman breast tumour (MDA‐MB‐231) xenograft nude mice, human cardiac ventricle fibroblasts, and human umbilical vein endothelial cells (HUVEC) were employed in the present study. Histology, immunohistochemistry, immunofluorescence, western blot, antibody array, and RNA‐sequencing analyses were utilized to assess the protective effect of Rh2 on cardiotoxicity induced by Dox and the underlying mechanisms.ResultsRh2‐reduced cardiotoxicity by inhibiting the cardiac histopathological changes, apoptosis and necrosis, and consequent inflammation. Pathological remodelling was attenuated by reducing fibroblast to myofibroblast transition (FMT) and endothelial–mesenchymal transition (EndMT) in hearts. RNA‐sequencing analysis showed that Dox treatment predominantly targets cell cycle and attachment of microtubules and boosted tumour necrosis, chemokine and interferon‐gamma production, response to cytokine and chemokine, and T cell activation, whereas Rh2 regulated these effects. Intriguingly, Rh2 also attenuated fibrosis via promoting senescence in myofibroblasts and reversing established myofibroblast differentiation in EndMT.ConclusionsRh2 regulates multiple pathways in the Dox‐provoked heart, proposing a potential candidate for cancer supplement and therapy‐associated cardiotoxicity.

Doxorubicin is extensively reported to induce severe cardiotoxicity in clinical applications. Our work proposed a natural herbal compound, ginsenoside Rh2, as a potential candidate for attenuating this side effect. Rh2 significantly inhibited cardiac apoptosis and necrosis, inflammation, and pathological remodelling in Dox‐challenged hearts.     

Reconstruction of Surface Porous PEEK Decorated with Strontium-doped Calcium Phosphate Coatings for Enhancing Osteogenic Activity

The aim of this study was to reconstruct surface porous structure with hundreds of micrometers and then bio-mineralize Sr-doped Calcium Phosphate(Sr-doped CaP)o...     

溶藻弧菌群体基因组学研究

溶藻弧菌(Vibrio alginolyticus)是一种能够对人类以及鱼、虾、贝类等水产品致病的弧菌,给人类健康带来威胁,也给水产养殖业造成巨大的经济损失.目前该物种基于全基因组的遗传多样性和重要遗传元件研究报道较少.本研究对采集自全国4个省份的68株溶藻弧菌进行高通量测序,获得全基因组序列,并结合113株公开发表的...     

三峡水库神农溪2014年春季浮游藻类演替成因分析

摘要：【目的】研究三峡水库神农溪库湾春季水华期间浮游藻类演替及其成因分析。【方法】2014年3–5月在神农溪库湾布置了6个断面(SN01–SN06),在神农溪汇入长江干流河口附近水域设置1个断面CJBD,对浮游藻类、相关环境因子及水动力因子进行了同步监测,据此分析了水体层化结构及水动力特性。【结果】神农溪在监测时段内共检测到浮游藻类6门38种(属);库湾浮游藻类生物量时间上差异显著(ANOVA,P<0.05)。春季浮游藻类群落结构具有明显的演替规律,3月份暴发大面积的硅藻水华（藻密度>100×105 cells/L),小环藻(Cyclotella spp.)为优势藻种;4月在SN02–SN06暴发以小球藻(Chlorella spp.)为主要优势种、衣藻(Chlamydomonas spp.)为次优势种的绿藻水华(藻密度>100×105 cells/L),5月份受水位大幅消落影响,浮游藻类生物量降低且无明显优势藻种。【结论】在具备充足的营养盐的水体中,水体层化结构与水动力特性对浮游藻类演替影响重大。三峡水库水位处于快速消落阶段时,流速成为抑制神农溪库湾藻类生长的主要因素。     

Development of High‐Level Omega‐3 Eicosapentaenoic Acid (EPA) Production from Phaeodactylum tricornutum

Phaeodactylum tricornutum is a lipid‐rich marine diatom that contains a high level of omega‐3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA). In an effort to reduce costs for large‐scale cultivation of this microalga, this study first established a New BBM medium (0.3 x strength BBM with only 3% of the initial phosphate level) to replace the traditional F/2 medium. Phaeodactylum tricornutum could grow in extremely low phosphate concentrations (25 µM), without compromising the EPA content. In the presence of sea salts, silicate addition was not necessary for high rate growth, high EPA content, or lipid accumulation in this species. Using urea as the sole nitrogen source tended to increase EPA contents per dry biomass (by 24.7%) while not affecting growth performance. The use of sea salts, rather than just sodium chloride, led to significantly improved biomass yields (20% increase) and EPA contents of total fatty acid (46–52% increase), most likely because it supplied sufficient essential elements such as magnesium. A salinity level of 35 led to significantly higher biomass yields compared with 20, but salinity had no significant influence on EPA content. EPA became the dominant fatty acid with average levels of 51.8% of total fatty acids during the exponential growth phase at 20 ppt in New BBM medium with sea salts.     

小兴安岭大亮子河国家森林公园不同生境下土壤细菌多样性和群落结构

土壤细菌是森林生态系统的一个重要组成成分, 是生态系统中物质循环和能量流动的重要参与者, 细菌群落组成和生物多样性是反映土壤生态功能的重要指标。本文利用高通量测序技术分析了大亮子河国家森林公园内红松(Pinus koraiensis)林、落叶松(Larix gmelinii)林、蒙古栎(Quercus mongolica)林、枫桦(Betula costata)林、针阔混交林、灌木林和草甸等7种不同生境土壤细菌群落组成和多样性的差异性, 探讨该地区土壤细菌群落对不同生境的响应, 为地区森林生态系统的经营管理及生态系统稳定性的维护提供科学理论基础。在门的水平上, 各生境的细菌群落组成基本一致, 其中变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)和疣微菌门(Verrucomicrobia)在7种生境土壤中相对丰度均大于10.0%, 是细菌中的优势菌门。在属的水平上, 共测得245个菌属, 各样地共有属118个, 占总属数的48.2%, 占总相对丰度的97.8%; 优势菌属分别为Spartobacteria_ genera_incertae_sedis、Gaiella、Gp16、Gp4, 占总相对丰度的47.0%, Spartobacteria_genera_incertae_sedis在7种生境土壤中丰度均最高。7种生境下的土壤细菌多样性和土壤理化因子存在着显著的差异, 红松林的土壤细菌群落多样性和丰富度均高于其他生境。土壤pH是大亮子河森林公园影响土壤细菌多样性的关键因子。