蚯蚓中抗肿瘤蛋白组分的提取分离及其抗肿瘤活性

以赤子爱胜蚓为原料 ,通过蛋白质的丙酮沉淀和凝胶过滤 ,分离得到一组抗肿瘤活性蛋白成分 (简称蚯蚓抽提物 ) ;这组成分富含Fe、Zn、Cu以及Se等微量元素 ,蛋白含量为 6 0 4 3± 2 36 %(n =5 ) .体外实验中 ,通过MTT法和SRB法测定了蚯蚓抽提物对多种人癌细胞株 (HCT 116、SY5Y、K5 6 2、MGc80 3和HeLa)以及正常细胞株 (HEK2 93和COS 7)的抑制杀伤率 .结果表明 ,蚯蚓抽提物对癌细胞的杀伤有一定的选择性 ,受试人癌细胞达到 5 0 %生长抑制率所需作用浓度约 6 0～110mg L ;但是 ,10 0℃煮沸 5min后 ,该抑制活性完全消失 .通过纤维蛋白平板法 ,测得蚯蚓抽提物同时具有纤溶酶和纤溶酶原激活酶的活性 .体外测得丝氨酸蛋白酶抑制剂aprotinin和PMSF能显著抑制蚯蚓抽提物的细胞杀伤活性 .体内实验中 ,蚯蚓抽提物能有效延长荷瘤 (S180 )小鼠的生存时间 ,并使其身体机能得到明显改善 .腹膜内注射剂量为 2 8mg kg和 36mg kg时 ,生命延长率分别为135 3%和 12 3 5 % ,和标准药物 (环磷酰胺 )治疗后结果 (76 5 % )相比 ,有显著性差异     

Biofilm coupled with UV irradiation for phenol degradation and change of its community structure

The extensive use of phenol compounds and the inability to remove these compounds during wastewater treatment have resulted in the widespread occurrence of phenols in the natural environment. Phenols have been linked to serious risks to human and environmental health. Hence, the need to develop technologies that can effectively remove phenols from wastewater and source waters is a pressing challenge. In this study, light ceramic particles were immersed in activated sludge acclimated to degrade phenol, and microorganisms were allowed to attach to the particles surface to form biofilm. Then the ceramic particles with biofilm were moved into the photolytic circulating-bed biofilm reactor made of quartz glass, which was used for the degradation of phenol by three protocols: photolysis with UV light alone (P), biodegradation alone (B), and the two mechanisms operating simultaneously (photobiodegradation, P&B). The experimental results indicated that phenol removal rate was quickest by B experiment. However, P&B experiment gave more complete mineralization of phenol than that by other protocols. During P&B experiment, the microorganisms grown on porous ceramic carrier still kept the bioactivity degrading phenol, even under UV light irradiation. However, the dominant members of the bacterial community changed dramatically after the intimately coupled photobiodegradation, according to molecular biological analysis to the biofilm. Whereas Beijerinckia sp. was the dominant strain in the inoculum, it was replaced by Thauera sp. MZ1T that played a main role on degrading phenol during P&B experiment.     

APols-Aided Protein Precipitation: A Rapid Method for Concentrating Proteins for Proteomic Analysis

Amphipols (APols) are a newly designed and milder class of detergent. They have been used primarily in protein structure analysis for membrane protein trapping and stabilization. We have recently demonstrated that APols can be used as an alternative detergent for proteome extraction and digestion, to achieve a “One-stop” single-tube workflow for proteomics. In this workflow, APols are removed by precipitation after protein digestion without depleting the digested peptides. Here, we took further advantage of this precipitation characteristic of APols to concentrate proteins from diluted samples. In contrast with tryptic peptides, a decrease in pH leads to the unbiased co-precipitation of APols with proteins, including globular hydrophilic proteins. We demonstrated that this precipitation is a combined effect of acid precipitation and the APols’ protein interactions. Also, we have been able to demonstrate that APols-aided protein precipitation works well on diluted samples, such as secretome sample, and provides a rapid method for protein concentration.     

云南伊蚊属奈蚊亚属一新种及二新记录种记述

报道了云南省伊蚊属奈蚊亚属Aedes subgen. Kenknightia Reinert,1990至今所发现的种类,其中包括1新种类异形伊蚊Ae.（Ken.） dissmilierodes sp. Nov.和2种中国新记录,讨论了新种与近缘种的鉴别特征,简述了2种新记录的形态特征。新种模式标本保存于云南省疟疾防治研究所。     

Binding and Uptake of RGD-Containing Ligands to Cellular α v β 3 Integrins

The cyclic peptide, cRGDf[N(me)]V, binds to the α _v β ₃ integrin and can disrupt binding of the integrin to its natural ligands in the extracellular matrix. In this work, the ability of a water-soluble, fluorescently labeled variant of the RGD-containing peptide (cRGDfK-488) to bind to integrins on human umbilical vascular endothelial cells (HUVEC) and subsequently undergo endocytosis was characterized. This information was compared to the binding and uptake properties of an α _v β ₃ integrin-specific monoclonal antibody, LM609X. The specificity of the RGD-containing peptide is assessed by comparison with control peptide that does not bind to the α _v β ₃ integrin, cRADfK-488. Using a high purity construct, it is shown that the RGD ligand exhibits dissociation constants in the micromolar range whereas LM609X exhibits dissociation constants in the nanomolar range. However, the RGD ligand showed greater uptake following incubation at temperatures which permit endocytosis. A 7.4-fold increase in uptake of the RGD peptide was observed following a 1 h incubation with HUVEC at 37°C (an endocytosis permissive temperature), as compared to that at 4°C (an endocytosis prohibitive temperature). In contrast, only a 1.9-fold increase in cell-associated fluorescence was observed for similar incubations with LM609X. Results from fluorescence microscopy supports the notion that the RGD peptide is rapidly endocytosed at 37°C as compared to LM609X. These results are discussed with regard to previous work indicating that RGD ligands enter cells by integrin-independent pathways. These studies provide well-controlled measures of how RGD ligands stimulate endocytosis. This may be of considerable interest for intracellular delivery of ligand-associated drugs in anti-angiogenic applications.     

杜仲花粉壁发育的超微结构观察

利用透射电子显微镜和场发射扫描显微镜对杜仲花粉壁结构和发育进行了研究.结果显示,杜仲花粉在四分体末期形成原外壁,质膜随之出现波状样式,原基粒棒的形成在波状质膜的凹陷处积累,同时细胞中的核质类核仁向核膜外扩散,并进一步分布在质膜下;单细胞花粉早期其外壁结构(覆盖层、柱状层、基足层和外壁内层)已经分化完全,之后花粉内壁由萌发孔起始发育,然后全面增厚,至花粉成熟时萌发孔处的内壁变得很厚,并形成具有丰富径向微通道的结构,此时外壁内层崩溃解体.重点讨论了花粉外壁模式决定以及类核仁和花粉外壁形成之间的关系.     

银杏外种皮提取液对菜粉蝶幼虫的毒力研究

用银杏外种皮水煮提取液进行室内菜粉蝶毒力实验和田间药效测定试验。结果表明,相当于0.75g/ml的银杏外种皮提取液对菜粉蝶3龄幼虫室内浸虫试验,幼虫死亡率为76.6%,浸叶饲喂健虫的幼虫死亡率为63.3%。0.15g/ml银杏外种皮提取液对菜粉蝶3龄幼虫室内浸虫试验,幼虫死亡率为53.3%,浸叶饲喂健虫死亡率为43.3%。0.75g/ml的银杏外种皮提取液对菜粉蝶5龄幼虫室内浸虫试验,幼虫死亡率仅为33.3%。0.75g/ml银杏外种皮提取液对菜粉蝶3龄幼虫田间药效测定,其幼虫死亡率为64%,校正防治效果为63.6%。     

危害茄科蔬菜的外来入侵害虫——马铃薯甲虫

马铃薯甲虫是世界著名的入侵害虫,是我国的重要检疫性有害生物,主要危害马铃薯、茄子和西红柿等茄科蔬菜。该甲虫目前分布于美洲、欧洲和亚洲等30多个国家和地区,于1993年入侵我国新疆,到2010年扩散到36个县市,最前沿为木垒县。该文介绍了马铃薯甲虫的扩散历史、危害情况、发生规律以及防治措施,指出我国在西北方向遭受新疆马铃薯甲虫入侵内地威胁的同时,也在东北方向遭受俄罗斯滨海边区马铃薯甲虫对黑龙江及东北地区入侵的严重威胁,应加强防范。     

基于F—2群体的鸡重要生长性状遗传分析

采用F-2设计,以丝羽乌骨鸡(C系)为一亲本,分别与农大褐蛋鸡(B系),及法国明星肉鸡(A系)进行正反交,产生了包含A×C,C×A;B×C,C×B4个杂交组合的F2代群体,建立了可用于定位产肉及产蛋等性状QTL的资源群体.基于这一F-2群体,对体重及日增重等性状表型值进行分析,结果显示,亲本间均值差异显著,分离群体变异大,这组性状间有中等或较高的相关,各性状潜在的QTL座位数不超过10,所建资源家系能够满足QTL定位所需的样本数量,达到了预期的效果.     

小型猪急性心肌梗死后心力衰竭模型的构建

目的应用选择性冠状动脉前降支(LAD)球囊闭塞结合微血栓微球混悬液灌注方法造成心肌缺血坏死,探索建立稳定存活的小型猪急性心肌梗死(AMI)后心力衰竭(HF)动物模型。方法选择中国五指山小型猪18头,行冠脉造影后沿血管送球囊至LAD中段,依次扩张球囊阻断前向血流1、2、5 min,每次间隔60 s,然后扩张球囊堵闭血流120 min。再以4F导管超选LAD,行微血栓微球混悬液分次注入,间隔10 min重复注射,TIMI心肌灌注分级(TMPG)2级和左室舒张末压(LVEDP)15 mm Hg时停止注射,同时监测心电图及应用漂浮导管监测有创血流动力学参数。并行pigtail导管测量(LVEDP)的变化,待LVEDP稳定在15~18 mm Hg之间后结扎血管,并加压包扎。监测心肌坏死标志物(cTnI和CK-MB)变化。分别于制模前,制模后第1天、7天、14天行心脏超声检查,制模第14天复查有创血流动力学检查,并行心脏病理检查,认定和评价模型的成功率、稳定性和可重复性。结果制模14 d后共有15头小型猪成活,心电图、心肌坏死标记物、病理检查均符合AMI病理生理过程。其中14头小型猪达到动物模型标准【肺毛细血管楔压(PCWP)18 mmHg和心输出量(CO)下降30%以上】,模型成功率为77.78%。制模后第14天PCWP明显升高(P0.01),CO平均下降50.76%;左室射血分数(LVEF)明显降低(P0.01)。病理检查显示心肌梗死面积占左心室面积的25.4%~34.9%。结论球囊闭塞结合微血栓微球混悬液灌注构建小型猪急性心肌梗死后心力衰竭模型具有闭胸、高成功率、稳定和重复性好等优点,较药物、冠状动脉结扎和起搏诱导的心力衰竭模型更接近临床病理生理学特点。