三氟氯氰菊酯对棉铃虫神经细胞延迟整流钾电流的抑制作用

用膜片钳技术首次研究了三氟氯氰菊酯对离体培养的棉铃虫中枢神经细胞延迟整流钾通道电流的影响。结果表明,药物作用前有81%和39%的细胞的通道分别在-30 mV 和 -40 mV 激活（n=21）。三氟氯氰菊酯（10^-5 mmol/L）作用15 min后,有63%和38%细胞的通道分别在-40 mV 和 -50 mV 激活（n=8）;作用1 min后电流幅值明显降低,抑制率达到了37.7%（n=19）;加药后激活曲线明显左移且Vh 值变化显著,但k值没有明显变化。实验结果说明,三氟氯氰菊酯作用后,通道更容易激活,但显著抑制电流峰值,导致神经敏感性降低,棉铃虫中枢神经细胞钾通道也是拟除虫菊酯类药物的作用靶标之一。     

应用纤维素结合域标签构建谷氨酸棒状杆菌表达系统

为优化谷氨酸棒状杆菌表达系统的纯化工艺,合成里氏木霉的CBD基因,将其与谷氨酸棒状杆菌分泌表达载体pXMJ19-sp连接,构建以CBD为纯化标签的重组载体pXMJ 19-sp-CBD.在该载体中插入GFP基因并转化至谷氨酸棒状杆菌,可获得分泌表达融合蛋白GFP-CBD的重组菌.该菌经IPTG诱导后的发酵液在紫外灯下显示强烈的绿色荧光,重组蛋白的分泌表达量达200 mg/L.利用CBD标签对纤维素柱的可逆性吸附,可直接对谷氨酸棒状杆菌分泌到培养基中的重组蛋白进行纯化,从而简化工艺和降低成本,为工业化大生产奠定基础.     

改性聚二甲基硅氧烷 (iPDMS) 蛋白芯片在肿瘤相关抗原自身抗体筛查中的应用

肿瘤标记的快速筛查是临床早期诊断的难题。利用化学发光蛋白质芯片技术,对低丰度的肿瘤相关抗原的自身抗体进行高灵敏度的筛选,是一种有益的尝试。本研究首先将带烯烃末端的、引发聚合反应的表面引发剂加入到常规聚二甲基硅氧烷材料中,再通过热交联反应固定到聚二甲基硅氧烷的三维结构中,形成改性聚二甲基硅氧烷 (iPDMS)。为了使iPDMS材料具有抗蛋白质非特异性吸附的特性,在活性引发位点处通过表面引发的原子转移自由基聚合反应合成poly(OEGMA) 高分子刷。最后将20种肿瘤相关的抗原利用高通量喷点打印技术打印到芯片的特定区域,并组装成iPDMS芯片的48孔检测微孔板。对临床上常见的8种肿瘤患者血清进行分析,发现VEGFR1和VEGF121自身抗体对常见的8种肿瘤具有检测价值,有望成为肿瘤快速筛查的检测指标。     

StructureActivity Relationships of Antimicrobial Gallic Acid Derivatives from Pomegranate and Acacia Fruit Extracts against Potato Bacterial Wilt Pathogen

Bacterial wilts of potato, tomato, pepper, and or eggplant caused by Ralstonia solanacearum are among the most serious plant diseases worldwide. In this study, the issue of developing bactericidal agents from natural sources against R. solanacearum derived from plant extracts was addressed. Extracts prepared from 25 plant species with antiseptic relevance in Egyptian folk medicine were screened for their antimicrobial properties against the potato pathogen R. solancearum by using the disc‐zone inhibition assay and microtitre plate dilution method. Plants exhibiting notable antimicrobial activities against the tested pathogen include extracts from Acacia arabica and Punica granatum. Bioactivity‐guided fractionation of A. arabica and P. granatum resulted in the isolation of bioactive compounds 3,5‐dihydroxy‐4‐methoxybenzoic acid and gallic acid, in addition to epicatechin. All isolates displayed significant antimicrobial activities against R. solanacearum (MIC values 0.5–9 mg/ml), with 3,5‐dihydroxy‐4‐methoxybenzoic acid being the most effective one with a MIC value of 0.47 mg/ml. We further performed a structure–activity relationship (SAR) study for the inhibition of R. solanacearum growth by ten natural, structurally related benzoic acids.     

Comparative proteomics analysis of silkworm hemolymph during the stages of metamorphosis via liquid chromatography and mass spectrometry

The silkworm is a lepidopteran insect that has an open circulatory system with hemolymph consisting of blood and lymph fluid. Hemolymph is not only considered as a depository of nutrients and energy, but it also plays a key role in substance transportation, immunity response, and proteolysis. In this study, we used LC‐MS/MS to analyze the hemolymph proteins of four developmental stages during metamorphosis. A total of 728 proteins were identified from the hemolymph of the second day of wandering stage, first day of pupation, ninth day of pupation, and first day as an adult moth. GO annotations and categories showed that silkworm hemolymph proteins were enriched in carbohydrate metabolism, proteolysis, protein binding, and antibacterial humoral response. The levels of nutrient, immunity‐related, and structural proteins changed significantly during development and metamorphosis. Some, such as cuticle, odorant‐binding, and chemosensory proteins, showed stage‐specific expression in the hemolymph. In addition, the expression of several antimicrobial peptides exhibited their highest level of abundance in the hemolymph of the early pupal stage. These findings provide a comprehensive proteomic insight of the silkworm hemolymph and suggest additional molecular targets for studying insect metamorphosis.     

高稳定人胰岛素的晶体学研究:Ⅲ,A21-Asp突变体的结晶和初步晶体学分析

研究一系列有明确意义改性胰岛素的结构将有益于新型胰岛素分子的设计。经蛋白质工程制备的AspA21—人胰岛素(ADHI)基本保持了生物活力,而在酸性溶夜(pH3—4)中的稳定性比天然胰岛素在中性溶液中高5—10倍。本文报道ADHI的结晶、初步晶体学研究及中等分辨率衍射数据的收集。ADHI晶体属R3空间群:a_H=b_H=82.72埃,c_H=34.02埃。     

西藏米拉卡黑白仰鼻猴的栖息地变化

黑白仰鼻猴(Rhinopithecus bieti)分布在我国金沙江和澜沧江之间的横断山脉的一个狭小区域内(26°14′N-29°20′N,99°15′E-99°37′E),海拔分布范围2600(南部)-4200m(北部);目前大约有15群,约1700只。本文所研究的西藏米拉卡猴群分布在我国西藏藏族自治区芒康县的南部,数量约50只。基于野外调查和过去的报道,此地的暗针叶林和针阔叶混交林是猴群的适宜栖息地,而由于人口数量增加所导致的夏季牧场和农田面积不断扩增正在逐渐侵蚀着这里的暗针叶林。为了评估该猴群的栖息地现状、变化趋势和变化原因,我们通过野外调查工作,利用GIS和RS技术,分别解译了当地过去5年(1986-2006年)冬季的Landsat TM卫星影像,并对解译结果进行了分析和计算,得到了该猴群栖息地的主要结果有:1)2006年暗针叶林面积是13600hmP2P,夏季牧场面积是4900hmP2P,农田面积是3300hmP2P;2)在过去20年(1986-2006年),暗针叶林面积减少了15.5%(2500hmP2P),牧场面积增加了58.1%(1800hmP2P),农田面积增加了17.8%(500hmP2P);3)暗针叶林的斑块数量增加了75.6%,平均斑块面积下降了51.8%(从1986年的15.3hmP2P到2006年的7.4hmP2P),最大斑块指数下降了54.7%;景观丰富度并没有变化,但Shannon多样性指数和Shannon均匀度指数分别增加了4.0%;4)暗针叶林面积变化与当地的人口数量呈显著负相关(r=?1.000),而夏季牧场和农田面积分别和当地人口呈显著正相关(r=1.000)。表明黑白仰鼻猴米拉卡猴群栖息地的丧失和破碎化程度较为严重;栖息地丧失和破碎化是当地传统生产方式和人口增长共同作用的结果。     

芽孢杆菌B3的鉴定及其对常见污染真菌的拮抗作用

从平菇废弃培养料中分离获得一株细菌B3,通过对其进行形态学、培养特征比较研究和16SrDNA序列分析,鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。在与常见的三种食用菌污染真菌脉孢菌(Neurospora sp.)、木霉(Trichoderma sp.)及鬼伞(Coprinus sp.)分别进行对峙培养时,都有明显的抑菌作用,形成的抑菌带分别为0.38cm、0.57cm及0.56cm,而与平菇(Pleurotus ostreatus)、鸡腿菇(Coprinus comatus)及草菇(Volvariella volvacea)分别进行对峙培养时,除对草菇有明显的抑制作用外,对平菇和鸡腿菇的抑制作用很小。在混接B3菌株麸皮菌剂栽培料培养基上,B3菌株对三种污染真菌的抑制作用都很明显;可抑制木霉分生孢子形成、脉孢菌分生孢子萌发及鬼伞菌丝扩展;但不抑制平菇菌丝的生长。