Con A affinity glycoproteomics of normal human liver tissue

In order to establish the novel high throughput, high efficiency and Iow cost technological platform for the research of N-glycoproteomics, to resolve the significance of characteristic expression profile of glycoprotein and to find the proteins with biological functional importance, the glycoproteins with high-mannose core and the two antennary types were purified and enriched by the Con A affinity chromatography. Con A affinity protein expression profiles of normal human liver tissue were generated by using SDS-PAGE, two-dimensional electrophoresis (2-DE) followed by fast fluorescence staining based on multiplexed proteomics (MP) technology. 301 visible protein spots on the gel were detected and 85 of glycoproteins were further successfully identified via peptide mass fingerprinting (PMF) by a matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS/MS) and annotated to IPI databases. Identified glycoproteins definitely take part in the regulation of cell cycle and metabolic processes. The glycosylation sites were predicted with NetNGlyc 1.0 and NetOGlyc 3.1 software, meanwhile they were classified according to the geneontology methods. The construction of Con A affinity glycoprotein database of normal human liver tissue would contribute to the subsequent research.     

基于GIS的森林景观定量分类

以金沟岭林场为例,利用电子林相图和森林资源二类调查资料基础,借助GIS软件MapInfo和数字化仪跟踪矢量化,并运用景观生态学原理和方法,选取分维数、分离度斑块密度、景观优势度和均匀度等指标,对林场的景观格局进行了分析;同时采用类平均聚类法,选取分维数、分离度、斑块平均面积、最大斑块面积4个景观类型特征指数作为聚类分析的聚类因子,进行定量地划分森林类型,合理地区划森林经营类型,为优化景观水平配置、制定生态采伐规程、编制森林经营方案提供理论基础。     

土地利用变化对海南土壤水源涵养功能的影响

热带地区已有大量原始林转变为其他土地利用类型,影响了陆地生态系统的水源涵养功能.为了明确热带原始林转变为其他土地类型对土壤水源涵养功能的影响,在海南中部山区选择4种典型土地利用类型,包括林龄大于100年的原始林(VF)、10年生次生林(SF)、12年生槟榔林(AF)和35年生橡胶林(RF),评估土地利用变化对土壤持水性能和水源涵养功能指数(SWI)的影响.结果表明: 与原始林相比,表层土壤(0～10 cm)中,其他土地类型土壤持水性能指标均降低,12年槟榔林各土层指标均最低.土壤含水量和最大持水量与植被郁闭度、土壤有机质和土壤容重显著相关,表明郁闭度、土壤有机质和紧实度的改变是土壤持水性能变化的重要原因.与原始林相比,次生林、槟榔林和橡胶林土壤水源涵养功能分别减少27.7%、54.3%和11.5%,不同土层的差异各异,橡胶林仅表层土壤水源涵养功能显著降低.植被郁闭度、土壤有机质和土壤容重可解释土壤水源涵养功能变量的83.3%.土地利用转变显著改变了土壤持水性能和土壤水源涵养功能,相比12年槟榔林,35年橡胶林能更好地保持土壤水分,土地管理中增加土壤有机质和减少土壤紧实度可改善土壤持水性能及水源涵养功能.     

Population control of the yellow‐spined bamboo locust,Ceracris kiangsu,using urine‐borne chemical baits in bamboo forest

The yellow‐spined bamboo locust, Ceracris kiangsu Tsai (Orthoptera: Oedipodidae), is a notorious defoliator of bamboos in China. In commercial bamboo forests, spraying insecticides to control C. kiangsu is neither convenient nor economic, therefore environmentally acceptable and cost‐effective methods are needed. Ceracris kiangsu adults are known to aggregate and gnaw at human urine‐contaminated materials; NaCl is a strong phagostimulant and NH₄HCO₃ is an attractant/arrestant. In the present paper, we found that foam plastic containers containing a blend of 0.03% bisultap, 3% NaCl, 3% NH₄HCO₃, and 0.1% Triton X‐100 (tetrad bait) had a powerful attracticidal effect and could kill a great number of especially female C. kiangsu adults, comparable to those containing a mixture of 0.03% bisultap and 5‐day‐incubated urine. In a field trial, the tetrad bait killed approximately 80% of females and about 9% of males in the treated plots after daily application for five consecutive days. The corrected average reduction rate was 48.2%. Moreover, the sex ratio was decreased from 1.18 before the trial to 0.25 after the experiment in the treated plots. These results indicated that there is the potential to develop a trap using NaCl as a phagostimulant and NH₄HCO₃ as an attractant that can be used for C. kiangsu control.     

Quality,bioactivity study,and preclinical acute toxicity,safety pharmacology evaluation of PEGylated recombinant human endostatin (M2ES)

Endostar, a potent endogenous antiangiogenic factor, is wildly used in clinics. However, it was easily degraded by enzymes and rapidly cleared by the kidneys. To overcome these shortcomings, PEGylated recombinant human endostatin was developed. In this study, the purity of M₂ES was evaluated by silver stain and reversed‐phase high‐performance liquid chromatography. Ultraviolet spectrum was used to examine the structural of M₂ES and endostar. The bioactivity and antitumor efficacy of M₂ES were evaluated using an in vitro endothelial cell migration model and athymic nude mouse xenograft model of a heterogeneous lung adenocarcinoma, respectively. A preclinical study was performed to evaluate the acute toxicity and safety pharmacology in rhesus monkeys. The purity of M₂ES was more than 98%; PEG modification has no effect on endostatin structure. Compared with the control group, M₂ES dramatically retards endothelial cell migration and tumor growth. After intravenous (IV) infusions of M₂ES at a dose level of three and 75 mg/kg in rhesus monkeys, there was no observable serious adverse event in both acute toxicity and safety pharmacology study. On the basis of the quality and bioactivity study data of M₂ES and the absence of serious side effect in rhesus monkeys, M₂ES was authorized to initiate a phase I clinical trial.     

飞机草和兰花菊三七光合作用对生长光强的适应

测定了干季不同光强下生长的飞机草和兰花菊三七叶片最大净光合速率(Pmax)、荧光动力学参数、叶绿素含量和比叶重(LMA),研究了两种植物适应光环境的策略,探讨了其与入侵性的关系.100%光强下,两种植物主要通过降低捕光色素复合体Ⅱ的含量减少光能吸收,提高Pmax增加光能利用维持叶片能量平衡,而它们的热耗散能力并不强,均显著低于其它光强下的值.100%光强下,提高类胡萝卜素含量是飞机草耗散过剩能量的有效策略,而兰花菊三七可能有其它耗散途径.4.5%光强下仅飞机草能够存活,它通过降低LMA、维持很低的日间热耗散和较高的光合系统Ⅱ非环式电子传递效率适应了弱光环境.推测对强光环境较强的适应能力是入侵植物的共性之一,但这种能力强不一定入侵性大.     

DNA损伤生物学反应中ATM对p21~(WAF1/CIP1)蛋白的直接磷酸化

毛细血管扩张性共济失调症突变蛋白 (mutatedinataxiatelangiectasia ,ATM)是直接感受DNA双链断裂损伤并起始诸多DNA损伤信号反应通路的主开关分子 .已有研究发现 ,DNA损伤生物学反应中 ,ATM可通过磷酸化活化p5 3,继而转录活化细胞周期检查点蛋白p2 1WAF1 CIP1的表达 ,而对于ATM是否直接参与p2 1WAF1 CIP1的早期活化迄今尚无实验证明 .通过免疫共沉淀反应 ,检测到细胞电离辐射 (ionizingradiation ,IR)反应早期ATM与p2 1WAF1 CIP1蛋白存在相互作用 .将p2 1WAF1 CIP1蛋白编码基因全长克隆入原核表达载体pGEX4T 2 ,经诱导表达及亲和层析纯化获取GST p2 1融合蛋白作为磷酸化底物 .体外磷酸化实验检测证明 ,IR活化的ATM具磷酸化p2 1WAF1 CIP1蛋白的功能 ,并且此磷酸化功能可被PI3K家族特异性抑制剂Wortmannin所抑制 .结果揭示了IR后ATM可通过直接磷酸化p2 1WAF1 CIP1蛋白 ,在IR致DNA损伤生物学反应早期调控p2 1WAF1 CIP1蛋白的快速活化过程     

阿维链霉菌中aveD基因缺失对阿维菌素合成的影响

利用aveD基因的缺失载体pCZ8(pKC1139∷△aveD)对阿维菌素(Avermectin)产生菌阿维链霉菌(Streptomyces avermitilis)76\|9的aveD基因进行缺失获得aveD缺失突变株。经摇瓶发酵和HPLC检测,发现该突变株只产生阿维菌素B组分。说明将阿维链霉菌的aveD基因缺失,并不影响下游aveF的表达。缺失突变株的阿维菌素的总产量与出发菌株的总产量基本相同,突变株中B1的产量略有提高,阿维菌素B2的含量显著提高。     

5406抗生菌肥特性及使用方法

5406菌肥的使用有多种方法,可与绿肥、厩肥混用或浸种、拌种、催芽和作为基肥使用等,都能起到很好的增产作用,如在缺磷地区使用,作物的增产效果更加明显。5406菌肥的浸出液也可以用于水稻与小麦浸种、浸根和喷施等,都能起到很好的增产效果。     

SARS病毒核蛋白基因的克隆及其表达研究

从一例输入性传染性非典型性肺炎病人血清中提取病毒RNA,通过RT-PCR方法扩增出SARS病毒核蛋白基因片段,克隆入质粒载体pUCm-T后,进行核苷酸序列的测定及分析,与已公布的SARS病毒基因序列进行比较,证实为SARS冠状病毒核蛋白基因.为了解该病毒核蛋白的抗原特性,将核蛋白基因插入表达载体,构建重组质粒pET28a-SN,转导大肠杆菌BL21(DE3)后,加IPTG诱导表达.产物经SDS-PAGE电泳分析,表达出相对分子量约为50kDa的蛋白,占整个菌体的45%左右.Western-blot分析表明,表达产物仅与SARS阳性病人血清起反应,而与正常血清不起反应.间接ELISA免疫检测,抗原滴度达112500.表明表达的核蛋白为SARS特异性抗原,这为SARS病毒的诊断试剂的研制提供了方便而安全的抗原来源.