Effects of cadmium on structure and enzymatic activity of Cu,Zn-SOD and oxidative status in neural cells

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder disease. Ten percent of the ALS patients are congenital (familial ALS), and the other 90% are sporadic ALS (SALS). It has been shown that mutations found in the Cu,Zn-SOD cause 20% of the familial ALS due to its low enzyme activity. We hypothesized that heavy metals may interfere the structure of Cu,Zn-SOD protein to suppress its activity in some of the SALS. In this study, we expressed and characterized the recombinant human Cu,Zn-SOD under various concentrations of Cu(2+), Zn(2+), and Cd(2+). By atomic absorption spectrophotometry, we demonstrated that adding of cadmium significantly increased the content of cadmium ion, but reduced its Zn(2+) content and enzyme activity of the Cu,Zn-SOD protein. The data of circular dichroism spectra demonstrated that the secondary structure of Cu,Zn-SOD/Cd is different from Cu,Zn-SOD, but close to apo-SOD. In addition to the effect of cadmium on Cu,Zn-SOD, cadmium was also shown to induce neural cell apoptosis. To further investigate the mechanism of neural cell apoptosis induced by cadmium, we used proteomics to analyze the altered protein expressions in neural cells treated with cadmium. The altered proteins include cellular structural proteins, stress-related and chaperone proteins, proteins involved in reactive oxygen species (ROS), enzyme proteins, and proteins that mediated cell death and survival signaling. Taken together, in this paper, we demonstrate that cadmium decreases the content of Zn(2+), changes the conformation of Cu,Zn-SOD protein to decrease its enzyme activity, and causes oxidative stress-induced neural cell apoptosis.     

阿氏浮丝藻mcyT基因序列多样性研究

为研究我国浮丝藻（Planktothrix Anagnostidis et Komrek）的毒素相关基因,选取分离自我国不同省份水体的13株阿氏浮丝藻,通过PCR检测其微囊藻毒素合成酶基因mcyA、mcyE及mcyT研究其毒素基因特性。PCR结果表明除mcyT之外其他引物检测均无扩增产物,说明这13株浮丝藻不具备产微囊藻毒素的能力。通过克隆测序得到mcyT序列,并进行分子系统分析,构建了关于mcyT序列的Neighbor-Joining系统树,结果表明mcyT序列可以将产毒与不产毒浮丝藻分为两大独立的分支,两个分支之间的最低序列相似度分别为98.5%和99.1%。研究结果可为后续研究我国浮丝藻的微囊藻毒素合成相关基因的多样性以及分子监测提供参考。       

大鳄龟感染蛙病毒的PCR检测及组织病理分析

2013年3月成都市某海洋馆送检2只发病大鳄龟(Macrochelys temminckii),临床特征表现为:精神状态萎靡,爬行无力,对外界刺激反应迟钝;颈部和四肢局部红肿,腹甲溃烂,严重部位甚至穿孔,最后死亡。为明确患病大鳄龟的病因,进行了细菌学、组织病理学和PCR检查。细菌学检查阴性;病理组织学观察发现,大鳄龟多组织、器官均发生严重病变,尤其是肾、肝、肺和心的损伤最为严重,表现为明显的变性、坏死和炎症细胞浸润,并在一些病变组织细胞浆内见嗜酸性或嗜碱性包涵体。针对蛙病毒(Ranavirus)病毒的特异性PCR检测扩增出蛙病毒主要衣壳蛋白(MCP)基因500 bp目的片段,测序后的DNA序列与Gen Bank中已知核酸序列进行Blast比对,发现其与Gen Bank中的蛙病毒主要衣壳蛋白基因同源性达95%~99%。根据组织病理特点及PCR检测结果推测大鳄龟的死亡是感染蛙病毒所致。     

Spatio-temporal properties of 5-lipoxygenase expression and activation in the brain after focal cerebral ischemia in rats

The role of 5-lipoxygenase (5-LOX) in brain injury after cerebral ischemia has been reported; however, the spatio-temporal properties of 5-LOX expression and the enzymatic activation are unclear. To determine these properties, we observed post-ischemic 5-LOX changes from 3 h to 14 days after reperfusion in rats with transient focal cerebral ischemia induced by 30 min of middle cerebral artery occlusion. We found that the expression of 5-LOX, both mRNA and protein, was increased in the ischemic core 12-24 h after reperfusion, and in the boundary zone adjacent to the ischemic core 7-14 days after reperfusion. The increased 5-LOX was primarily localized in the neurons in the ischemic core at 24 h, but in the proliferated astrocytes in the boundary zone 14 days after reperfusion. As 5-LOX metabolites, the level of cysteinyl-leukotrienes in the ischemic brain was substantially increased 3 h to 24 h, near control at 3 days, and moderately increased again 7 days after reperfusion; whereas the level of LTB(4) was increased mildly 3 h but substantially 7-14 days after reperfusion. Thus, we conclude that 5-LOX expression and the enzymatic activity are increased after focal cerebral ischemia, and spatio-temporally involved in neuron injury in the acute phase and astrocyte proliferation in the late phase.     

酚酸和氮素交互作用下欧美杨107细根形态特征

树木细根生长与根际过程的关系十分密切。该研究仿生欧美杨107 (Populus × euramericana ‘Neva’)人工林根际土壤酚酸沉降与氮素有效性变化, 通过设置3种酚酸梯度(0X、0.5X、1.0X, X为田间土壤酚酸含量)与3种氮素水平(缺氮0 mmol·L^-1、正常氮10 mmol·L^-1、高氮20 mmol·L^-1), 探究酚酸和氮素对欧美杨107细根形态的影响, 以期为阐明树木根系生长对根-土界面过程的响应奠定基础。结果表明: (1)在无酚酸(0X)环境中, 缺氮和高氮均可抑制欧美杨107细根生长, 尤其对1-3级细根的影响更为显著。比根长随氮素水平升高逐渐减小, 但其他细根特征并未呈现与氮素水平的线性关系。(2) 0.5X和1.0X酚酸梯度相比, 欧美杨107的1-2级细根直径和体积随酚酸浓度增加而显著增大(p < 0.05)。酚酸和氮素对杨树细根的影响存在交互作用, 1-2级细根直径、体积受酚酸的影响显著, 而4-5级细根长度、表面积受氮素影响显著。双因素方差分析结果表明, 酚酸和氮素对细根形态建成具有协同或拮抗效应。(3)主成分分析(PCA)和冗余分析(RDA)结果表明, 在酚酸和氮素交互效应下, 杨树1-3级、 4级、 5级细根之间具有显著的形态差异。第一主成分主要体现细根觅食性状特征, 可解释细根形态变异的60.9%的信息; 第二主成分主要体现细根形态构建特征, 可解释25.3%的信息。杨树细根形态变化与根序高度相关, N素影响杨树细根形态的主效应较酚酸更强。因此, 根际环境中酚酸累积和氮素有效性变化会影响杨树细根的形态构建和细根对水分、养分的吸收, 而氮素有效性是影响杨树细根生长的重要因素, 开展杨树人工林土壤养分管理是林分生产力长期维持的关键。     

The design, synthesis and biological evaluation of 7-alkoxy-4-heteroarylamino-3-cyanoquinolines as dual inhibitors of c-Src and iNOS

Because both c-Src and iNOS are key regulatory enzymes in tumorigenesis, a new series of 4-heterocycle amine-3-quinolinecarbonitriles as potent dual inhibitors of both enzymes were designed, synthesized and evaluated as multiple targets agents in cancer therapy. All compounds were evaluated by two related enzyme inhibition assays and an anti-proliferation assay in vitro. The results showed that most compounds inhibited c-Src and iNOS well. The best compound 8 inhibited both enzymes with the IC(50) values of 34.8 nM and 26.7 microM. Several compounds also showed moderate anti-proliferation at 10 microM against colon and liver cancer cell lines.     

水果湖湾蓝藻水华的形成及其对东湖影响的评价

东湖是武汉市最重要的湖泊,位于湖北省武汉市武昌区,属中型浅水湖泊,它集饮用水源、旅游、疗养、气候调节、水产养殖和水上运动等多功能于一体[1].因此,东湖水质的好坏与武汉人民的生活密切相关.