脱氧鬼臼毒素对美洲大蠊背侧不成对中间神经元电压

目的:探讨脱氧鬼臼毒素(DOP)对美洲大蠊背侧不成对中间神经元(DUM)电压依赖性钾电流IK的影响。方法:采用全细胞膜片钳技术研究脱氧鬼臼毒素对美洲大蠊背侧不成对中间神经元电压依赖性钾电流的电流幅度,电流-电压关系以及激活曲线的影响。结果:DOP能够抑制电压依赖性钾通道电流的幅度,而且此抑制作用具有浓度依赖性(5、10、20、40μmol/L)。DOP抑制IK的半数抑制浓度(IC50)值为18.064μmol/L。20μmol/L DOP能使IK的电流-电压关系曲线下移,并能使IK的激活曲线向去极化方向移动。结论:DOP对美洲大蠊背侧不成对中间神经元(DUM)电压依赖性钾电流具有抑制作用,这可能是其杀虫作用的机制之一。     

脱氧鬼臼毒素对美洲大蠊的毒力及几种酶系的影响

采用药膜法测试了脱氧鬼臼毒素对美洲大蠊Periplaneta americana初孵若虫的触杀活性,并测定了其对成虫中枢神经系统乙酰胆碱酯酶（AChE）和腺苷三磷酸酶(ATPase)离体活性的影响。结果表明：脱氧鬼臼毒素对美洲大蠊初孵若虫具有较强的毒杀活性,在接触时间24、48、72和96 h 的LC₅₀分别为26.26、4.68、1.51和0.62 μg/cm²;其对AChE没有明显影响; 对Na⁺ -K⁺ -ATPase有明显抑制作用,并存在浓度 效应关系,IC₅₀为44.9 μmol/L; 对Ca²⁺-Mg²⁺-ATPase表现出低剂量激活,高剂量抑制的现象。结果提示美洲大蠊的AChE不是脱氧鬼臼毒素靶标,而ATPase可能是脱氧鬼臼毒素的重要靶标之一。     

脱氧鬼臼毒素对美洲大蠊乙酰胆碱受体信号通路分子mRNA表达水平的影响

【目的】探讨脱氧鬼臼毒素(deoxyopodophyllotoxin, DOP)对美洲大蠊Periplaneta americana神经系统乙酰胆碱受体信号通路的影响。【方法】从美洲大蠊头部克隆乙酰胆碱受体信号通路上的关键信号分子nAChR α6亚基、 CaM和 CaMKⅡ的部分mRNA, 并测定其序列。应用荧光定量PCR技术分别观察注射不同浓度（10, 45, 80, 115和150 μg/μL） DOP 24和48 h后上述3种基因表达水平的变化。【结果】测序结果显示, 克隆出的美洲大蠊nAChR基因部分序列（539 bp）与赤拟谷盗Tribolium castaneum nAChR α6亚基基因的核苷酸序列一致性为84%; 美洲大蠊CaM基因（435 bp）与雕叶蝉Graphocephala atropunctata CaM基因的核苷酸序列一致性为85%; 美洲大蠊CaMKⅡ基因（513 bp）与黑腹果蝇Drosophila melanogaster CaMKⅡ基因的核苷酸一致性为77%。实时定量荧光PCR实验表明： DOP处理48 h后对美洲大蠊nAChR α6亚基、 CaM和CaMKII基因表达水平大体表现出低剂量激活, 高剂量抑制的特点。45~80 μg/μL DOP浓度范围内3种基因表达水平达到高峰, 80~150 μg/μL 浓度范围内表现为抑制作用, 基因表达水平呈下降趋势。【结论】DOP需要在美洲大蠊体内蓄积一定时间才有明显的作用, 能与nAChR结合引起CaM-CaMKⅡ级联反应, 使3种基因的表达在低浓度组上调、 高浓度组被抑制, 进而对美洲大蠊产生潜在的毒杀作用。     

五种昆虫多糖的体外抗氧化活性

采用邻苯三酚自氧化法、DPPH法、Fenton反应体系、FRAP分析法及对过氧化氢损伤细胞保护作用方法对白蜡虫Ericerus pela Cavannes雌成虫多糖、蝗虫多糖、喙尾琵甲多糖、美洲大蠊多糖及家蚕蛹多糖进行体外抗氧化研究,评价昆虫多糖总还原能力,对超氧阴离子自由基(O2-·)、DPPH·、羟自由基(·OH)的清除作用及损伤细胞存活率影响。试验结果表明,5种多糖对几种自由基均有不同程度的清除作用,且均呈现一定的量效关系;但对不同的氧化体系呈现不同的清除活性,白蜡虫多糖的总还原能力最强;白蜡虫、喙尾琵甲、美洲大蠊及蝗虫多糖对DPPH的清除率相对较明显;在羟自由基的清除能力上蚕蛹、喙尾琵甲及美洲大蠊多糖相对较强;各昆虫多糖对超氧阴离子清除能力不明显。5种多糖对过氧化氢损伤细胞有一定保护作用,在不同浓度范围内能增加细胞存活率。研究表明昆虫多糖具有一定抗氧化活性,可成为天然抗氧化物的来源。     

四种蜚蠊的呼吸信号特征比较

在20℃下,利用CO₂红外分析仪采集了美洲大蠊Periplaneta americana、褐斑大蠊P.brunnea、澳洲大蠊P.australasiae和德国小蠊Blattella germanica的呼吸信号。结果表明,它们均具有典型的不连续气体交换循环（discontinuous gas exchange cycle, DGC）呼吸模式,且一个DGC可分为爆发间期和爆发期2个阶段。4种蜚蠊雄性成虫DGC的特征各异：美洲大蠊完成一个DGC周期平均约需24.55 min,明显长于褐斑大蠊(11.67 min)和澳洲大蠊(10.75 min),德国小蠊周期最短,仅为4.41 min;对于爆发间期历时在整个DGC历时所占的比例,美洲大蠊最大,平均为57％,德国小蠊次之,为48％,褐斑大蠊和澳洲大蠊比较接近,分别为37％和36％。德国小蠊单位体重的CO₂平均释放速率最大,而另外3种蜚蠊的差异不明显。4种蜚蠊爆发期CO₂释放体积均随DGC历时增加而增加,美洲大蠊和德国小蠊单位体重的CO₂平均释放率随DGC历时增加而减少,在褐斑大蠊和澳洲大蠊中它们关系不明显。美洲大蠊、褐斑大蠊和澳洲大蠊CO₂平均释放率和爆发期CO₂释放体积与体重呈正相关,在德国小蠊中关系不明显;4种蜚蠊DGC各阶段历时和DGC频率与它们体重的关系均不明显。     

灭多威对美洲大蠊腹六神经节突触传递的阻断作用

用电生理学方法研究了灭多威对美洲大蠊Periplanetaamerwana腹六神经节(A6节)突触传递的影响。用灭多威溶液浸泡A6节,电刺激尾须神经粗支,用甘露醇间隙法记录兴奋性突触后电位(EPSP)和突触后动作电位。给予弱刺激只记录到EPSP时,灭多威作用初期EPSP幅度增加、时程延长,能诱发突触后动作电位,随后EPSP逐渐减小至消失,冲洗可恢复,突触前反应保持不变。增加电刺激强度记录到突触后动作电位时,灭多威可阻断A6节的突触传递,阻断时间是浓度依赖性的,阻断是可逆的,但冲洗30 min仍保留一定的后作用。对美洲大蠊雄性成虫腹腔注射灭多威测定致死中量(LD₅₀)为(3.56±0.01) μg/g体重。根据灭多威的作用机理对其阻断A6节突触传递的特点以及对虫体的毒杀机制进行了讨论。     

杀虫环对黑胸大蠊神经突触传递的阻遏作用

用电生理糖间隙法研究杀虫环对黑胸大蠊神经突触传递的作用,并以α-银环蛇毒素作比较。结果证明:1)杀虫环阈浓度1×10^-5M即显著地抑制兴奋性突触后电位(EPSP)。作用开始使之阈值递增,此时只有增加刺激强度方可诱出EPSP。2)(虫非)蠊第Ⅵ腹神经节是胆碱能的。已知突触后阻遏剂如α-银环蛇毒素的作用是N型乙酰胆碱受体(n-AchR)的专一性配基,与杀虫环阻遏神经突触的传递颇为相似,二者均不影响突触后神经元的静息电位和动作电位的传导;而杀虫环对非胆磁能的神经肌肉接头则无影响。3)自发突触后电位随杀虫环处理时间的不同而变化。开始自发释放电位的振幅、频率逐渐增加,继之产生持续期较长的阵发性高频发放,以后又逐渐消失。     

美洲大蠊粪便粗提物的聚集活性

用不同溶剂不同方法提取美洲大蠊Periplaneta americana(L.)粪便,测定美洲大蠊各虫态的诱集活性。结果表明,采用直接浸泡提取方法,丙酮、乙醇、正己烷和二氯甲烷4种粗提物对美洲大蠊各虫态都具有明显的诱集作用,其中乙醇和丙酮粗提物的引诱效果最好,正己烷次之,二氯甲烷最弱。4种溶剂粗提物对美洲大蠊雄成虫和高龄若虫聚集活性最强,对低龄若虫聚集活性最弱。用乙醇溶剂对粪便粗提,3种提取方法均对美洲大蠊有很强的诱集效果,其中索氏抽提诱集效果最弱,直接浸泡和超声波提取效果好,且差异不显著,但直接浸泡提取效果更好。     

鬼臼毒素对小菜蛾的生物活性及对其几种代谢酶系的影响

为进一步揭示鬼臼毒素的杀虫活性及作用机理,本实验采用小叶碟添加法测定了鬼臼毒素对小菜蛾Plutella xylostella的生物活性及对其幼虫体内羧酸酯酶（CarE）、酸性磷酸酯酶（ACP）、碱性磷酸酯酶（AKP）、谷胱甘肽S-转移酶（GSTs）和细胞色素P450酶系活性的影响。结果表明：鬼臼毒素对小菜蛾具有较好的拒食、毒杀及生长发育抑制作用,其AFC₅₀为0.4110 mg/mL（24 h）和0.2617 mg/mL（48 h）,LC₅₀为1.9061 mg/mL（72 h）;0.125 mg/mL剂量下相对生长率比对照下降57.56％（24 h）和41.18％（48 h）,化蛹率为43.33％,不能正常羽化。同时,鬼臼毒素对小菜蛾幼虫体内各代谢酶存在不同程度的影响：对羧酸酯酶表现为先激活后抑制的作用;对酸性磷酸酯酶表现先抑制后激活的作用; 对碱性磷酸酯酶活性具有明显的抑制作用,且随着处理时间的延长,抑制作用增强;对谷胱甘肽S-转移酶和细胞色素P450酶系表现出先激活后抑制的影响。     

美洲大蠊成虫肠道抗细菌共生放线菌的筛选及鉴定

【目的】鉴于野外美洲大蠊Periplaneta americana对恶劣环境适应性强,本研究旨在从云南省大理州的野外美洲大蠊成虫肠道中分离、筛选出抗细菌活性放线菌,为抗生素开发提供菌种资源。【方法】采用涂布平板法和平板划线法对美洲大蠊成虫肠道放线菌进行分离;以金黄色葡萄球菌Staphylococcus aureus、耐甲氧西林金黄色葡萄球菌、铜绿假单胞菌Pseudomonas aeruginosa、粪肠球菌Enterococcus faecalis、大肠杆菌Escherichia coli和鼠伤寒沙门氏菌Salmonella typhimurium共6种人体病原细菌为指示菌株,采用牛津杯法对分离自这些放线菌的次生代谢产物进行抗菌活性测定;通过形态学特征和16S rRNA基因序列分析,对具有广谱和明显抗菌活性的放线菌进行鉴定,并经16SrRNA基因序列的BlAST同源性比对及系统发育分析确定它们的分类地位。【结果】从美洲大蠊成虫肠道共分离获得41株放线菌。抗菌活性测定结果表明,34株(82.9%)放线菌对至少1种指示病原细菌具有抑制作用,其中有7株对3种以上病原细菌具有抑制作用,9株表现...     

Ultrasound-induced changes in synaptic processes with different transmitters in smooth muscles

The effect of therapeutic-intensity ultrasound on neuromuscular transmission and spontaneous electrical and contractile activity in smooth muscles of the gastrointestinal tract of guinea pig was studied by a modified sucrose-gap technique. The action of ultrasound was found to facilitate the acetylcholinergic neuromuscular transmission (mainly by increasing the amplitude of excitatory postsynaptic potentials). The higher efficiency of the nonadrenergic neuromuscular transmission was manifested as an increase (nearly twofold) in the total duration, but not in the amplitude, of inhibitory postsynaptic potentials. Modulations of the first and second components of the potentials caused respectively by the action of ATP and of nitric oxide as possible transmitters, were different. Concurrently with enhancing the synaptic transmission efficiency, ultrasound exerted an opposite, inhibitory, effect on generation of spontaneous action potentials and contraction of smooth muscles. All the ultrasound effects were fully reversible. The findings permit assuming a special mechanism of modification of the synaptic transmission in smooth muscles under the action of ultrasound.Neirofiziologiya/Neurophysiology, Vol. 25, No. 4, pp. 297–302, July–August, 1993.     

Spontaneous Action Potential Initiation and Propagation in Regenerating Cell Segments of Acetabularia mediterranea

Thavarungkul, P., Lertsithichai, S. and Sherlock, R. A. 1987.Spontaneous action potential initiation and propagation in regeneratingcell segments of Acetabularia mediterranea.—J. exp. Bot.38: 1541–1556. The spontaneous action potentials which occur during the regenerationof anucleate isolated stalk segments (ISS's) of the unicellularalga Acetabulana mediterranea have been studied using a novelnon-invasive technique. This involved measurement of spatialsamples of the time dependent potential in the external medium(sea water) and an inverse transformation to give the currentdistribution at the cell surface. The initiation region andsubsequent propagation (if any) of each action potential wasthen analysed from the computed transmembrane currents. Theresults showed that the occurrences of the spontaneous actionpotentials followed a rhythm which had a period of approximately24–30 h. These action potentials initiated more frequentlyat the ends of the ISS's than in the middle region. Our resultsshowed no firm correlation between the regions of action potentialinitiation and the site of the regenerating apex. Generallyboth propagating and non-propagating action potentials wereobserved in the same ISS, the ratio of which seems to dependon the length of the cell. The analysed initiation sites andthe propagating behaviour of the spontaneous action potentialgenerated by the ISS's during the regeneration process do notprovide any clear support to the symmetry breaking role of thepropagating action potentials as proposed by some workers. Key words: Acetabularia, regeneration, action potential     

Inotropism and Contracture of Aplysiid Ventricles as Related to the Action of Neurohumors on Resting and Action Potentials of Molluscan Hearts

The postsynaptic actions of neurohumors on molluscan musclemay be exerted through control of force as well as by meansof excitation and inhibition. The control of force may appearas potentiation of the response to excitation, as increasedinotropism in spontaneous contractions, or as an increase intonus. We have directed our attention to the alterations offorce induced in aplysiid ventricles by applied postulated neurohumors.The results are interpreted in terms of the known effects ofneurohumors on resting potential and action potentials of molluscanhearts. We recorded compound membrane potentials of aplysiidventricles extracellularly, using a single sucrose gap apparatustogether with a force-displacement transducer to measureforcein contractions or contracture. Aplysia californica ventriclehas a membrane potential of –52.5 ± 9.4 mV. Ventriclesof Aplysia dactylomela or Aplysia californica are depolarizedby increased concentrations of external potassium ion, withan accompanying contracture. After incubation in calcium-freemedium, KCl contracture-force is directly dependent on calciumion concentration. A depolarization of 8.3 ±2.14 mV inpotassium-free medium is blocked by substitution of lithiumfor sodium in the medium, suggesting an electrogenic sodiumpump. There is a sustained depolarization in low chloride medium,which suggests a significant chloride contribution to the restingpotential. Ventricles of Dolabella auriculana or Aplysia dactylomelaare depolarized by acetylcholine (ACh). Threshold for depolarizationis lower than threshold for contracture-force. The ventricleof A. dactylomela is depolarized by 5- hydroxytryptamine (5HT)with threshold at 10^–9 M and a maximum depolarizationof 30 mV at 10^–4M. Depolarization by 5HT may induce beatingbut does not induce contracture. Ventricles of Aplysia californicaare not depolarized by ACh although beating ventricles are inhibited,and a depolarized ventricle in a tonic contracture may be hyperpolarizedand relaxed by low concentrations. The force of contractionof the ventricle of Dolabella auricularia is dependent on theduration of the plateau phase of the cardiac action potential.The plateau is lengthened by 5HT with an accompanying increasein force of beat, and shortened by ACh, with an accompanyingdecrease in force of beat. The action potential in the ventricleof Aplysia californica is not differentiated into spike andplateau phases, and neither ACh nor 5HT has any marked effecton the form of the action potential. Nevertheless, isolatedventricles of Dolabella auricularia, Aplysia dactylomela, andAplysia californica are all excited by 5-hydroxytryptamine,with a threshold at about 10^–M. Both spontaneous beatingand excitation induced in A. californica ventricles by 5HT areblocked by lack of the sodium ion, which may be responsiblefor pacemaker potentials in molluscan hearts.     

甲醚菊酯和溴氰菊酯在果蝇幼虫外周神经系统的协同毒理作用

本文运用细胞内微电极记录技术研究了甲醚菊酯和溴氰菊酯对果蝇Drosophila melanogaster幼虫神经一肌肉突触兴奋性接点电位(EJP₅)的影响。用甲醚菊酯(1.49x10^-8m01/L)处理后引起果蝇EJP₅的自发释放增加和刺激后的重复后自发释放。而用溴氰菊酯(1.0x10^-8mol/L)处理的则无明显影响。这显示甲醚菊酯对果蝇外周神经主要为I型毒理作用。而溴氰菊酯则主要为Ⅱ型作用,甲醚菊酯和溴氰菊酯联合应用后,则产生兼具I型和II型特征的自发释放或诱发EJP₅发放。自发释放或重复后自发释放的频率和幅值随联合处理中甲醚菊脂和溴氰菊酯的配比而变化。这些结果说明甲醚菊酯和溴氰菊酯对果蝇幼虫外周神经的毒理具有协同作用。     

Effect of Folate Analogues on the Activity of Dihydrofolate Reductase and Callus Growth of Sunflower

The inhibitory effect of antifolate analogues in vitro and invivo on dihydrofolate reductase and on growth of sunflower callus,was tested. Aminopterin and trimethoprime proved to be the mosteffective inhibitors. The ID₅₀ values for these compounds werethe same as those previously shown for enzymes from rice andcarrot cells. Protein, DNA and RNA synthesis of sunflower callus was inhibitedby the analogues, the inhibition depending on the concentrationused. These results suggest that growth inhibition of sunflowercallus by these analogues is due to inhibition of dihydrofolatereductase. Key words: Antifolate, callus, dihydrofolate reductase, growth, sunflower     

Changes of action potentials and force at lowered [Na+]o in mouse skeletal muscle: implications for fatigue

We examined 1) whether the effects of lowered trans-sarcolemmal Na⁺ gradient on force differed between nonfatigued fast- and slow-twitch muscles of mice and 2) whether effects on action potentials could explain the decrease of force. The Na⁺ gradient was reduced by lowering the extracellular [Na⁺] ([Na⁺]_o). The peak force-[Na⁺]_o relationships for the twitch and tetanus were the same in nonfatigued extensor digitorum longus and soleus muscles: force was maintained over a large range of [Na⁺]_o and then decreased abruptly over a much smaller range. However, fatigue was significantly exacerbated at a lowered [Na⁺]_o that had little effect in nonfatigued soleus muscle. This finding suggests that substantial differences exist in the Na⁺ effect on force between nonfatigued and fatigued muscle. The reduced contractility in nonfatigued muscles at lowered [Na⁺]_o was largely due to 1) an increased number of inexcitable fibers and threshold for action potentials, 2) a reduction of action potential amplitude, and 3) a reduced capacity to generate action potentials throughout trains. sodium gradient; muscle contraction; action potential train; extensor digitorum longus; soleus     

The effects on stretching and prostaglandin F2alpha on the contractile and bioelectric activity of the uterus in rat.

The effects of stretching and of prostaglandin F2alpha on spontaneous and induced with local deformation contractile activity of rat uterus were studied. It was found that stretching the uterus up to the length of the decontracted organ in vivo increased the contractile and bioelectric activity. The rise in spontaneous uterine activity was a factor reducing induction of additional contractions. The fall in the level of endogenous prostaglandins in the uterus following administration of indomethacin inhibited the spontaneous contractile activity but was without effect on the contractions induced by local deformation of the myometrium. Prostaglandin F2alpha added to the bath in a concentration of 0.001 ng/ml exerted an inhibitory action on the different tested parameters of the contractile activity. After high doses stimulation of the uterine activity was observed.     

The Effects of Temperature on Action Potentials in the Chill Sensitive Seismonastic Plant Biophytum sensitivum

The seismonastic plant Biophytum sensitivum was stimulated electricallyto produce reproducible action potentials. It was found thatnegative voltages only induced a response and that stimulationcould also be achieved by electrostatic means by depositingnegative charge on the leaf surface. A study of the effectsof temperature on the propagation rates of action potentialsin different aged leaves showed that for leaves of all agesthe fastest propagation occurred at 24 ?C. As the temperaturewas decreased or increased from 24 ?C a reduction in the propagationrate occurred. No propagation was found at, or below, 14 ?Cand the propagation rate decreased with increasing leaf age.Both the threshold voltage level and the charge required toproduce action potentials in young leaves was lowest at 24 ?Cand had to be increased at either lower or higher temperaturesto cause leaf closure. The duration of the electrical stimulusrequired to induce action potentials was found to decrease asthe voltage was increased. Mechanisms which could account forthe effects of temperature on action potentials are discussedin relation to the chill-sensitivity of the plant. Key words: Chilling, Action Potentials, Biophytum sensitivum     

（－）SPD和（－）THP对蓝斑核去甲肾上腺素能神经元自发放电的影响

利用在体记录大鼠蓝斑核神经元单位放电,研究了（－）ＳＰＤ和（－）ＴＨＰ对其放电活动的影响。结果表明：（－）ＳＰＤ通过去甲肾上腺素α２受体,以剂量依赖方式增强蓝斑核神经元放电,但较大剂量却对神经元放电有一定抑制。然而（－）ＴＨＰ可使蓝斑核去甲肾上腺素能神经元出现可逆性放电抑制。     

Synthesis and biological evaluations of oleanolic acid indole derivatives as hyaluronidase inhibitors with enhanced skin permeability

Oleanolic acid (OA) is a natural cosmeceutical compound with various skin beneficial activities including inhibitory effect on hyaluronidase but the anti-hyaluronidase activity and mechanisms of action of its synthetic analogues remain unclear. Herein, a series of OA derivatives were synthesised and evaluated for their inhibitory effects on hyaluronidase. Compared to OA, an induction of fluorinated (6c) and chlorinated (6g) indole moieties led to enhanced anti-hyaluronidase activity (IC₅₀ = 80.3 vs. 9.97 and 9.57 µg/mL, respectively). Furthermore, spectroscopic and computational studies revealed that 6c and 6g can bind to hyaluronidase protein and alter its secondary structure leading to reduced enzyme activity. In addition, OA indole derivatives showed feasible skin permeability in a slightly acidic environment (pH = 6.5) and 6c exerted skin protective effect by reducing cellular reactive oxygen species in human skin keratinocytes. Findings from the current study support that OA indole derivatives are potential cosmeceuticals with anti-hyaluronidase activity.