补骨脂素对TCP磨损颗粒所致大鼠成骨细胞损伤的干预作用及其机制

目的: 探讨补骨脂素(psoralen)对TCP磨损颗粒诱导成骨细胞损伤的影响及其分子机制。方法: 通过消化法从SD大鼠乳鼠颅骨中获取原代的成骨细胞,应用碱性磷酸酶(ALP)染色鉴定成骨细胞。TCP磨损颗粒(0.1 mg/ml)与成骨细胞共孵育48 h构建成骨细胞损伤的体外实验模型,实验随机分为正常对照组(Control)、模型(TCP)组和psoralen(10^-7 mol/L、10^-6 mol/L和10^-5 mol/L)组。WST法和流式细胞术分别检测各组成骨细胞活性变化和凋亡情况;化学比色法检测成骨细胞中ALP活性;各组成骨细胞培养14 d后应用茜素红S染色观察矿化结节形成。Western blot法检测各组成骨细胞中葡萄糖调节蛋白78/94(GRP78/94)、肌醇依赖酶1α(IRE1α)、剪切型X盒结合蛋白1(XBP1s)和磷酸化c-Jun氨基末端激酶(p-JNK)等蛋白质的表达。结果: 与Control组比较,TCP组成骨细胞活性、ALP活性和矿化结节的生成显著降低(P＜0.05),细胞凋亡率、GRP78/94、IRE1α、XBP1s和p-JNK等蛋白质表达明显增加(P＜0.05);与TCP组比较,补骨脂素各组成骨细胞损伤情况明显减轻,细胞凋亡率显著减少(P＜0.05),GRP78/94、IRE1α、XBP1s和p-JNK等蛋白质表达也明显下降(P＜0.05)。结论: 补骨脂素可抑制TCP磨损颗粒诱导的IRE1α-XBP1s-JNK信号通路的激活,阻止TCP颗粒所致的成骨细胞损伤及凋亡。     

A型塞内卡病毒的病毒样颗粒的制备及其免疫原性分析

为了研制A型塞内卡病毒 (Senecavirus A,SVA) 的病毒样颗粒 (Virus-like particles,VLPs) 疫苗,以SVA田间流行毒株CH-FJ-2017结构蛋白基因序列为研究对象,构建了能够同时表达SVA的3种结构蛋白VP0、VP1和VP3的单个原核重组表达质粒pET28a-SVA-VP031。通过大肠杆菌Escherichia coli表达、亲和层析纯化和体外自组装,获得SVA VLPs。透射电子显微镜鉴定显示,SVA的3种结构蛋白在体外能够自组装成直径约25–30 nm的VLPs,并且动物免疫试验结果表明,该VLPs能够有效刺激豚鼠产生高水平的抗原特异性中和抗体。上述研究结果为SVA VLPs疫苗的研制奠定了基础。     

中国西北地区天然胡杨群体遗传多样性及核心保护单元的构建

胡杨(Populus euphratica)是极端干旱荒漠区的珍稀乔木树种。为了确定天然胡杨群体遗传多样性保护单元并挖掘优异的种质资源, 本研究以中国西北地区新疆、青海、甘肃、宁夏、内蒙古的58个天然胡杨群体为研究对象, 利用120个位点的SNPs标记对这些胡杨群体进行群体遗传结构和遗传多样性分析, 并根据不同群体间Nei’s遗传相似度, 采用逐步聚类优先取样法对初始群体、遗传多样性保护单元和剩余群体进行t检验。群体结构和主成分分析表明, 胡杨群体可分为新疆南疆(SX)、新疆北疆(NX)、青海(QH)和混合群(甘肃、宁夏和内蒙古混合群, GNM) 4个分支, 遗传多样性分析表明新疆南疆(SX)遗传多样性高于其他群体。分子方差分析(AMOVA)表明天然胡杨群体的遗传变异主要分布在各个群体内。构建了天然胡杨群体一级核心保护单元3个群体(CU3), 二级核心保护单元33个群体(CU33)。南疆存在较多优异抗逆的天然胡杨古树资源, 南疆分布区的平均遗传多样性水平最高。综上所述, 南疆地区胡杨古树遗传多样性整体高于北疆及疆外地区, 结合新疆地区干旱严重指数等生境信息, 建议加大对南疆胡杨古树群体的保护力度, 重视北疆胡杨林的更新换代。     

Suppression Of Hepatitis C Virus Core Protein By Short Hairpin Rna Expression Vectors In The Core Protein Expression Huh-7 Cells

Short hairpin RNAs (shRNAs) efficiently inhibit gene expression by RNA interference. Here, we report the efficient inhibition by DNA-based vector-derived shRNAs of core protein expression in Huh-7 cells. The shRNAs were designed to target the core region of the hepatitis C virus (HCV) genome. The core region is the most conserved region in the HCV genome, making it an ideal target for shRNAs. We identified an effective site on the core region for suppression of the HCV core protein. The HCV core protein in core protein-expressing Huh-7 cells was downregulated by core protein-shRNA expression vectors (core-shRNA-452, 479, and 503). Our results support the feasibility of using shRNA-based gene therapy to inhibit HCV core protein production.     

我国小豆应用核心种质的生态适应性及评价利用

为提高我国小豆(Vigna angularis)应用型核心样本的育种利用效率,本研究通过两年两点的田间试验,分析了166份这些核心种质农艺性状的变异情况,明确了其生态适应性和可利用价值。结果表明,166份小豆核心种质在广西南宁均表现为直立生长,并能正常成熟收获,;在河北唐山仅有64.2%XX88份种质的小豆种质在河北唐山两年内均表现为直立生长,且有25份不能正常开花结荚。大部分数量性状年度间差异与生态环境有关,其中唐山的年度间差异不明显,而南宁的年度间均达到极显著差异,反映了小豆对生态环境、气候条件的较强敏感性。所有种质在南宁的生育期显著提前,而产量相关因子中,单株荚数、百粒重等均显著小于唐山。综合分析表明不同种质的生态适应性存在较大差异,说明选育广适性小豆品种是可能的。综合最终根据农艺性状的综合表现,分别筛选出适合南宁和唐山育种利用的优异种质共16份。筛选出适合南宁种植的绿豆种质8多少份,适合唐山种植的绿豆种质10多少份。     

Exploring core competencies for mental health and addictions work within a Family Health Team setting

Approximately 200 Family Health Teams (FHTs) have been implemented in Ontario to improve access to primary healthcare, including mental health and addiction. The objectives of this project were to examine, through a focus group and qualitative methodology with three FHTs, the profile of patients'' mental health and addiction-related needs and to identify the implications for the development of core competencies in these innovative organisations. A spectrum of needs and service trajectories was identified, as well as the importance of a wide range of clinical skills and knowledge. The results indicate that ‘core’ competencies for mental health work in the context of an FHT go well beyond those required for an embedded mental health ‘programme’ or specialised mental health counsellors, but rather they relate to the core and discipline-specific competencies of members of the entire team. In addition to specific knowledge and skills, competencies include common attitudes and values relating to teamwork, good communication and collaboration. Challenges were noted with regard to working with some community service providers, especially addiction services. Implications for core competencies at the individual and organisational level were identified.     

Acceptor specificities and selective inhibition of recombinant human Gal- and GlcNAc-transferases that synthesize core structures 1, 2, 3 and 4 of O-glycans

Background

Modifications of proteins by O-glycosylation determine many of the properties and functions of proteins. We wish to understand the mechanisms of O-glycosylation and develop inhibitors that could affect glycoprotein functions and alter cellular behavior.

Methods

We expressed recombinant soluble human Gal- and GlcNAc-transferases that synthesize the O-glycan cores 1 to 4 and are critical for the overall structures of O-glycans. We determined the properties and substrate specificities of these enzymes using synthetic acceptor substrate analogs. Compounds that were inactive as substrates were tested as inhibitors.

Results

Enzymes significantly differed in their recognition of the sugar moieties and aglycone groups of substrates. Core 1 synthase was active with glycopeptide substrates but GlcNAc-transferases preferred substrates with hydrophobic aglycone groups. Chemical modifications of the acceptors shed light on enzyme–substrate interactions. Core 1 synthase was weakly inhibited by its substrate analog benzyl 2-butanamido-2-deoxy-α-d-galactoside while two of the three GlcNAc-transferases were selectively and potently inhibited by bis-imidazolium salts which are not substrate analogs.

Conclusions

This work delineates the distinct specificities and properties of the enzymes that synthesize the common O-glycan core structures 1 to 4. New inhibitors were found that could selectively inhibit the synthesis of cores 1, 2 and 3 but not core 4.

General significance

These studies help our understanding of the mechanisms of action of enzymes critical for O-glycosylation. The results may be useful for the re-engineering of O-glycosylation to determine the roles of O-glycans and the enzymes critical for O-glycosylation, and for biotechnology with potential therapeutic applications.     

Design and simulation of a micro-channel for separating the particles with nearly constant dielectrophoretic force in channel space

One of the common approaches for separating the micro-particles is the utilisation of dielectrophoresis (DEP) force. In the present study, a channel structure is introduced in which the controllable electric field is imposed by the use of various voltages to the parallel electrodes in order to finally move the different particles in two opposite sides of the channel. The simulation results reveal that the channel is characterised by the imposition of nearly constant DEP forces in the whole channel, which is the innovation of this work. The samples used for separation are Latex particles with 216 and 557 nm in diameter.     

Appendant structure plays an important role in amyloidogenic cystatin dimerization prior to domain swapping

It has been hypothesized that prior to protein domain swapping, unfolding occurs in regions important for the stability of the native monomeric structure, which probably increases the possibility of intermolecular interaction. In order to explore the detailed information of the important unfolding regions in cystatin prior to domain swapping, 20?ns molecular dynamic simulations were performed at atomic level with typical amyloidogenic chicken cystatin (cC) mutant I66Q monomer under conditions that enable forming amyloid fibrils in biological experiments. Our results showed that I66Q mutant exhibited relatively large secondary structure changes and obvious expanding tendency of hydrophobic core compared to wild-type cC. More importantly, the appendant structure (AS) showed a large displacement and distortion towards the hydrophobic core in amyloidogenic cystatin. The structural analysis on cystatin monomer suggested that structural changes of the AS might make the hydrophobic core expand more easily. In addition, analysis on docking dimer has shown that the distorted AS was favor to intermolecular interactions between two cystatin monomers. Data from an independent theoretical derived algorithm as well as biological experiments also support this hypothesis.     

Affinity of Placental Decorin for Collagen

Decorin was isolated from 7 M urea extract of bovine placental cotyledons by ion-exchange and hydrophobic chromatography. Decorin and its core protein showed a broad band at about 115 kDa and a single band at 47 kDa, respectively by SDS-PAGE. Anti-decorin core protein antiserum from pig skin was reacted with placental decorin and its core protein in western blotting. The NH₂-terminal amino acid sequence of core protein from placental cotyledons was not different from that of core protein from skin and bone. Glycosaminoglycan of decorin was identified as dermatan sulfate by electrophoresis on a cellulose-acetate membrane and chondroitinase digestivity. Decorin bound to collagen in the order for type III, I, and V.