荧光增白剂Tinopal LPW对蜀柏毒蛾核型多角体病毒增效作用研究

用1%TinopalLPW荧光增白剂作为蜀柏毒蛾核型多角体病毒增效剂对蜀柏毒蛾2龄幼虫进行室内毒力测定,结果表明1%Tinonal LPW对Parocneria orienta NPV有较强的增效作用.使用3.6×1011PIB/hm2+1%TinopalLPW、1.8×1011PIB/hm2+ 1%TinopalLPW、9.0×1010PIB/hm2+1%TinopalLPW和3.6×1011PIB/hm2、1.8×1011PIB/hm2、9.0×1010PIB/hm26种处理对林间越冬代2-3龄幼虫进行超低容量喷雾防治,结果表明除9.0×1010PIB /hm2+1%TinopalLPW 表现出显著的增效作用外,其余剂量有增效作用但不显著.     

荧光增白剂UBL对蜀柏毒蛾核型多角体病毒增效作用研究

用1%UBL荧光增效剂作为蜀柏毒蛾核型多角体病毒的增效剂,对蜀柏毒蛾2龄幼虫进行室内毒力测定,结果表明1%UBL对Parocneria orienta NPV有较强的增效作用。     

蜀柏毒蛾核型多角体病毒的分离鉴定

本文报道了新分离的一株核型多角体病毒:蜀柏毒蛾核型多角体病毒(Paroceneria orient Nuclear Polyhedrosis Virus)。其多角体为四边形、五边形、大小在1.06—2.42μm。病毒粒子杆状,大小为385×55nm。室内感染蜀柏毒蛾幼虫其死亡率达9S％具较强的毒力。     

柏毒蛾核型多角体病毒的分离鉴定

柏毒蛾核型多角体病毒的分离鉴定李崇荣,彭辉银,周显明,陈新文,谢天恩（贵州省铜仁地区林科所,铜仁５５４３００）（中国科学院武汉病毒研究所,武汉４３００７１）（贵州省林业科学研究院,贵阳５５００１１）关键词柏毒蛾,核型多角体病毒柏毒蛾（Ｐａｒｏｃｅｎｅ...     

蜀柏毒蛾核型多角体病毒结构多肽及基因组酶切分析

对蜀伯毒蛾核型多角体病毒（Ｐａｒｏｃｎｅｒｉａ ｏｒｉｅｎｔａ Ｎｕｃｌｅａｒ ｐｏｌｙｈｅｄｒｏｖｉｒｕｓ,简称ＰａｏｒＮＰＶ）形态结构、结构多肽、限制内切酶图谱等特性进行了研究。采用不连续系统垂直板ＳＤＳ－ＰＡＧＥ分析了ＰａｏｒＮＰＶ的多角体蛋白、病毒粒子结构多肽。应用５种限制性内切酶对ＰａｏｒＮＰＶ基因组ＤＮＡ进行了酶切分析。结果表明：经热处理的多角体蛋白仅有一条带,分子量为３１．５ｋＤ,不     

蜀柏毒蛾核型多角体病毒结构多肽及基因组酶切分析

对蜀柏毒蛾核型多角体病毒(Parocneria orienta Nuclear polyhedrovirus,简称PaorNPV)形态结构、结构多肽、限制性内切酶图谱等特性进行了研究.采用不连续系统垂直板SDS-PAGE分析了PaorNPV的多角体蛋白、病毒粒子结构多肽.应用5种限制性内切酶对PaorNPV基因组DNA进行了酶切分析.结果表明:经热处理的多角体蛋白仅有一条带,分子量为31.5 kD,不经热处理的多角体蛋白有三条带,分子量分别为31.5 kD、29.1 kD、28.6 kD;病毒粒子包含有25种结构多肽,分子量范围在17.6-114.6 kD之间.PaorNPV DNA经BamH I.EcoR I、HindⅢ、Pst I和Xho I酶切分别产生9、12、12、12和14条片段.基因组大小平均为124.6 kb.     

蜀柏毒蛾核型多角体病毒基因文库的构建及几丁质酶基因的定位

蜀柏毒蛾 (ＰａｒｏｃｎｅｒｉａｏｒｉｅｎｔａＣｈａｏ)是柏木、桧柏、干头柏等柏科树种的重要食叶害虫[1] ,到目前为止仅存在于我国。蜀柏毒蛾核型多角体病毒(Ｐａｒｏｃｎｅｒｉａｏｒｉｅｎｔａｎｕｃｌｅａｒｐｏｌｙｈｅｄｒｏｖｉｒｕｓ ,ＰａｏｒＮＰＶ) [2 ]于 1991年被分离 ,该病毒对蜀柏毒蛾幼虫具有较强毒力 ,已作为一种新型的生物农药初步应用于柏木林区害虫的防治上[3] 。对这种病毒已进行了一些研究 ,包括生物活性测定 ,形态结构 ,理化特性 ,限制性内切酶分析[4 ,5] 。为了进一步研究其分子生物学特性 ,开发我国这种特有的…     

蜀柏毒蛾核型多角体病毒基因文库的构建及几丁质酶基因的定位

蜀柏毒蛾(Parocneria orienta Chao)是柏木、桧柏、干头柏等柏科树种的重要食叶害虫[1],到目前为止仅存在于我国.蜀柏毒蛾核型多角体病毒(Parocneria orienta nuclear polyhedrovirus,PaorNPV)[2]于1991年被分离,该病毒对蜀柏毒蛾幼虫具有较强毒力,已作为一种新型的生物农药初步应用于柏木林区害虫的防治上[3].对这种病毒已进行了一些研究,包括生物活性测定,形态结构,理化特性,限制性内切酶分析[4,5].为了进一步研究其分子生物学特性,开发我国这种特有的病毒资源,我们构建了部分PaorNPV基因组DNA片段的基因文库,同时以中国棉铃虫单粒包埋核型多角体病毒(HearSNPV)几丁质酶基因作探针,对PaorNPV几丁质酶基因进行了定位,结果报道如下.     

杉叶毒蛾核型多角体病毒的分离鉴定

杉叶毒蛾 (Ｄａｓｙｃｈｉｒａｔｈｗａｉｔｅａｉ) ,又名大茸毒蛾 ,属鳞翅目 ,毒蛾科[1] 。在我国广东 ,广西及西南等省区均有分布 ,贵州省全省杉木产区均有发生 ,主要危害杉木、茶、油茶、枫香和华山松等 ,1 995年于贵州省黄平采集到自然病死幼虫 ,经分离鉴定 ,该病原物为一种核型多角体病毒。1　材料与方法1.1　ＤｔＳＮＰＶ多角体的分离纯化取自然病死虫体充分捣碎 ,以 50 0ｒ/ｍｉｎ离心 5ｍｉｎ ,去除细胞碎片和沉淀 ,取上清于 4 0 0 0ｒ/ｍｉｎ离心 30ｍｉｎ。用蒸馏水悬浮沉淀 ,重复上述方法差速离心 2次 ,所得多角体较为纯净 ,置冰…     

荧光素对舞毒蛾核型多角体病毒不同地理品系的增效与光保护作用

舞毒蛾Lymantria dispar L.是源于欧亚大陆的多食性叶部害虫,取食300多种乔灌木,现已分布于北美、北非,成为世界性危险害虫之一,给林业生产带来了巨大损失。舞毒蛾核型多角体病毒(LdMNPV)是控制舞毒蛾种群动态的重要生物因素,可引起舞毒蛾种群急剧下降。在室内采用青杨枝条饲养的方法,测定了来自中国、北美和日本的3个LdMNPV品系(分别为LdMNPV-H,LdMNPV-D和LdMNPV-J)对危害青杨的亚洲型舞毒蛾幼虫的毒力,并测定了荧光素Tinopal LPW对它们的增效和光保护作用。结果表明Tinopal LPW对LdMNPV 3个地理品系均有增效和光保护作用,而且随着Tinopal LPW浓度的增加,增效作用增强,1%Tinopal LPW的增效作用最好。添加1%Tinopal LPW的LdMNPV-D品系、LdMNPV-H品系和LdMNPV-J品系对取食青杨的舞毒蛾幼虫的致死中浓度(LC50)分别为1.0、1.6、17.6 OBs/μL,不添加1%Tinopal LPW时,它们的LC50分别为32.9、39.0、1076.4 OBs/μL,分别降低了33、24、61倍。不添加1%Tinopal LPW时,D、H和J品系对舞毒蛾二龄幼虫的致LC95分别是2125.5、1275.8、303540.0 OBs/μL,添加1%Tinopal LPW后LC95分别为73.0、285.4、2360.8OBs/μL,分别降低了26、4.5、128.6倍。此外,1%Tinopal LPW的荧光素使3个品系的致死中时间(LT50)分别缩短了2.9d、5.3d、1.2d。LdMNPV-D和LdMNPV-H品系对亚洲型舞毒蛾表现出低致死中浓度、较短的致死中时间和较大的斜率,二者的毒力较LdMNPV-J品系高,在生产实践中应选择LdMNPV-D添加1%Tinopal LPW。Tinopal LPW对LdMNPV-D、LdMNPV-H和LdMNPV-J 3个品系均有光保护作用,添加1%Tinopal LPW后在距离30W紫外灯40cm下照射16h后,它们毒力保持系数比未添加Tinopal LPW分别高1.8、2.6、1.8倍。     

Use of a Stilbene Brightener,Tinopal LPW,as a Radiation Protectant for Steinernema carpocapsae

A stilbene fluorescent brightener, Tinopal LPW, was used as an ultraviolet (UV) protectant for the entomopathogenic nematode Steinernema carpocapsae (All strain). Irradiation of an aqueous suspension of nematodes produced a LC₅₀ in 15.7 minutes under a sunlamp and in 31.7 minutes in direct sunlight. Irradiation by both sunlamp and sunlight of a suspension of nematodes in Tinopal LPW did not reduce their biological activity as measured by their ability to parasitize wax moth larvae after exposure of 8 hours and 4 hours, respectively. Tinopal LPW appeared promising as a radiation protectant.     

Effects of 2,6-dichlorobenzonitrile and Tinopal LPW on the structure of the cellulose synthesizing complexes ofVaucheria hamata

Summary The effects of 2,6-dichlorobenzonitrile (DCB, a known inhibitor of cellulose synthesis) and Tinopal LPW (TPL, an agent which disrupts glucan crystallization) on the structure of cellulose synthesizing complexes (terminal complexes, TCs) in the xanthophycean algaVaucheria hamata were investigated. DCB (10 M) inhibits nascent fibril formation from the TC subunit (based on the absence of impressions) although it does not alter the overall shape of the rectangular TC during the short treatment of 20 min. With a prolonged treatment (60 min), the arrangement of TC subunits becomes disordered, and particles generally exhibited as doublets of subunits are released from each other. DCB also interferes with the formation of the overall shape of the TC although it does not disturb the conversion into TC rows of the subunits (the zymogenic precursor of the TC) packed in the globules. A 15 min treatment with TPL (1 mM) destroys the TC integrity by reducing the subunits into small fragments or particulate aggregates. The particulate rows of the TC are interrupted at many points, and fragments and particulate aggregates are dispersed by prolonged treatment (45 min) with TPL. Unlike DCB, TPL inhibits the conversion of globule subunits into TC rows. New insights on the structural characteristics necessary for cellulose microfibril assembly and possible mechanisms for the biogenesis of theVaucheria TC come from these data.Abbreviations DCB 2,6-dichlorobenzonitrile - TPL Tinopal LPW - TC terminal complex     

Interference of cell wall regeneration ofBoergesenia forbesii protoplasts by Tinopal LPW,a fluorescent brightening agent

Summary Wounding cells ofBoergesenia forbesii (Harvey) Feldmann induces the synchronous formation of numerous protoplasts which synthesize large cellulose microfibrils within 2–3 hours after wounding. The microfibrils appear to be assembled by linear terminal synthesizing complexes (TCs). TC subunits appear on both E- and P-faces of the plasma membrane, thus suggesting the occurrence of a transmembrane complex. The direction of microfibril synthesis is random during primary wall assembly and becomes ordered during secondary wall assembly. The average density of TCs during secondary wall deposition is 1.7/m², and the average length of the TC is 510 nm. TC organization is similar to that ofValonia macrophysa; however, the larger TCs ofBoergesenia (510 nm vs. 350 nm) produce correspondingly larger microfibrils (30 nm vs. 20 nm).The effects of a fluorescent brightening agent (FBA), Tinopal LPW, on cell wall regeneration ofBoergesenia protoplasts was investigated. The threshold level of Tinopal LPW for interfering with microfibril assembly is 1.5 M. At 95 M Tinopal (for short periods up to 15 minutes), microfibril impressions have atypical spherical impressions at their termini. At longer incubations (24 hours), TCs and microfibril impressions are absent. When washed free of Tinopal, the protoplasts eventually resume normal wall assembly; however, TCs do not reappear until at least 30 minutes after the removal of Tinopal. In consideration of the presence of ordered TCs before FBA treatment, their random distribution upon recovery implies an intermediate stage of assembly or possiblyde novo synthesis.     

Laboratory and field studies on the reproductive ecology of Parocneria orienta (Lepidoptera: Lymantriidae) in China

To lay a foundation for the monitoring, prevention and control of Parocneria orienta, a pest moth in China, we investigated the reproductive ecology of P. orienta, including the environmental factors affecting reproduction and the effects of pheromones in field traps. The results showed that the mating duration is shorter when the temperature is higher, with the optimal temperature range for P. orienta reproduction being 22–27°C. The sex ratio had an effect on the male mating capacity, which significantly increased when the sex ratio was ♀:♂ ≥ 3:1. The average number of matings was 2.44 for males, while female P. orienta mated only once in a lifetime. Males were more attracted to virgin females than to mated females, and the temperature and light period had an effect on the release of sex pheromones by females. Field trap tests using virgin females supported the results obtained in the laboratory, showing that the release of sex pheromones by P. orienta follows a distinct circadian rhythm.     

Synergistic effects of nisin and thymol on antimicrobial activities in Listeria monocytogenes and Bacillus subtilis

Nisin Z and thymol were tested, alone and in combination, for antibacterial activity against Listeria monocytogenes ATCC 7644 and Bacillus subtilis ATCC 33712. The antibacterial effect of nisin Z, produced by Lactococcus lactis KE3 isolated from the traditional Moroccan fermented milk, was greatly potentiated by sub-inhibitory concentrations of thymol in both bacterial strains. Our data showed that the concentration of nisin required for effective control of food-borne pathogenic bacteria could be considerably lowered by the use of thymol in combination. The use of low concentrations of nisin could lead to a less favourable condition for the occurrence of nisin-resistant bacterial sub-populations.     

Synergistic anti-glioma effect of Hydroxygenkwanin and Apigenin in vitro

Apigenin (AP) and Hydroxygenkwanin (HGK) are two natural flavonoid compounds. Previous studies have already demonstrated the anti-tumor capability of AP. However, it is not clear whether HGK has such property. In the current study, the anti-glioma activities of HGK and its synergistic anti-glioma effects with AP on C6 glioma cells were investigated. In addition, the possible mechanisms were also studied.     

Synergistic effect of Se-methylselenocysteine and vitamin E in ameliorating the acute ethanol-induced oxidative damage in rat

The present study was conduced to investigate the synergistic effects of combined treatments with Se-methylselenocysteine (SeMSC) and vitamin E (Vit E) in reversing oxidative stress induced by ethanol in serum and different tissues of rats. Sixty female rats were randomly divided into six groups for 30 days’ consecutive pretreatments as followed: control (I), physiological saline (II), 2.8 μg kg⁻¹ Se as SeMSC (III), 2.8 μg kg⁻¹ Se as sodium selenite (Na₂SeO₃, IV), 5 mg kg⁻¹ α-tocopherol as α-tocopherol acetate (Vit E, V), 5 mg kg⁻¹ α-tocopherol as α-tocopherol acetate and 2.8 μg kg⁻¹ Se as SeMSC (VI). All animals in groups II–VI were treated by ethanol treatment to cause oxidative stress. After 6 h of ethanol treatment, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), the contents of total antioxidant capacity (T-AOC), malondialdehyde (MDA), glutathione (GSH) and carbonyl protein (CP) in the serum, liver, heart and kidney were measured. The result showed that the individual SeSMC, Na₂SeO₃ and vitamin E could effectively increase the SOD, T-AOC, GSH-Px and GSH contents as well as significantly decrease the MDA and CP concentrations in the tissues of ethanol-induced rats. At the same dose on different forms of Se, SeMSC showed greater antioxidant activity than Na₂SeO₃. Moreover, group VI (SeMSC and α-tocopherol acetate) showed much better antioxidant activity than individual group III (SeMSC) and V (α-tocopherol acetate) due to the synergistic effect.     

阿帕替尼与白蛋白结合型紫杉醇在MDA-MB-231乳腺癌细胞系中的协同抗癌作用

目的阐明阿帕替尼 (apatinib)和白蛋白结合型紫杉醇 (nab-Paclitaxel)诱导MDA-MB-231乳腺癌细胞凋亡的分子机制。方法本研究以MDA-MB-231乳腺癌细胞为研究对象,并以apatinib和nab-Paclitaxel处理细胞后分组:0.1 %DMSO处理为阴性对照组;10 μmol/L apatinib处理组 (APA组);经5、10、15、20 nmol/L nab-Paclitaxel 处理组 (Nab-p 5组、Nab-p 10组、Nab-p 15组和Nab-p 20组);以及10 μmol/L apatinib分别与5、10、15、20 nmol/L nab-Paclitaxel 联合处理组 (APA+Nab-p 5组、APA+Nab-p 10组、APA+Nab-p 15组和APA+Nab-p 20组)。使用乳酸脱氢酶释放测定法测定apatinib和nab-Paclitaxel对MDA-MB-231细胞诱导的细胞毒活性,结合流式细胞术分析不同处理组细胞凋亡情况,通过JC-1染色法测定不同干预方式对MDA-MB-231细胞线粒体膜电位变化 (ΔΨ_m)的影响,借助FAM-FLICA荧光成像检测caspase-8和caspase-9 活性,通过彗星试验评估apatinib和nab-Paclitaxel对非致瘤上皮细胞株MCF-10A细胞DNA损伤的影响。两组之间独立样本t检验或单向ANOVA进行比较,多组之间使用Tukey事后检验。结果细胞毒性检测结果显示,与阴性对照组相比,Nab-p 20组MDA-MB-231细胞杀伤率在24 h时接近90 %;与单药处理组 (Nab-p 5组和Nab-p 10组)相比,APA+Nab-p 5组和APA+Nab-p 10组联合处理24 h和48 h后,分别检测到约85 %和95 %细胞死亡,差异均具有统计学意义 (P 均 < 0.001)。流式细胞术统计结果显示,与Nab-p5组和Nab-p10组相比,APA+Nab-p 5组和APA+Nab-p 10组24 h时MDA-MB-231细胞凋亡率(31.8 %±1.48 %、33.25 %±1.77 %比76.11 %±1.14 %、89.4 %±1.07%)升高 (P 均 < 0.05)。线粒体膜电位检测结果表明,与对照组相比,在单药处理组中,仅APA组和Nab-p 10组24 h时去极化细胞 (12.35 %±1.05%比78.33%±1.11%、46.74%±1.75%)增多;在联用处理组中,APA+Nab-p 5组和APA+Nab-p 10组24 h时去极化细胞 (68.47%±1.94%比90.03%±1.79%)增多,差异均有统计学意义 (P 均 < 0.05)。FAM-FLICA荧光成像结果显示,相较于单一处理组,apatinib和nab-Paclitaxel联用处理组的caspase-8和caspase-9蛋白高度活化。结合彗星试验分析,apatinib和nab-Paclitaxel 干预对MCF-10A非致瘤上皮细胞株DNA完整性没有显著影响。结论 apatinib/nab-Paclitaxel联用通过内源性的线粒体功能扰动和外源性的caspase激活诱导三阴性乳腺癌细胞MDA-MB-231凋亡,发挥协同抗癌作用。