奈达铂与鼻咽癌相关蛋白P53、Bcl-2、caspase的表达水平与相关性分析

目的 观察奈达铂对鼻咽癌CNE-2细胞增殖和凋亡的影响,探讨奈达铂与鼻咽癌相关蛋白P53、B淋巴细胞瘤-2(Bcl-2)、caspase的表达水平与相关性分析.方法 分析不同浓度的奈达铂(0 μg/ml、5 μg/ml、10 μg/ml、20 μg/ml)和作用不同时间(12 h、24 h、36 h、48 h)对鼻咽癌CNE-2细胞增殖和凋亡的影响;分析不同浓度的奈达铂对CNE-2细胞P53、Bcl-2、caspase蛋白表达水平的影响.结果 使用MTT法检测不同浓度的奈达铂对CNE-2细胞增殖的影响,随奈达铂浓度的升高OD值逐渐降低(P<0.05),且随着奈达铂作用时间的延长OD值逐渐降低(P<0.05).形态学可见:奈达铂浓度0 μg/ml时CNE-2细胞增值迅速、细胞饱满,形态规则;奈达铂作用下细胞增值减慢,细胞变圆、边缘模糊;使用流式细胞仪检测不同浓度的奈达铂对CNE-2细胞凋亡率可见随着奈达铂浓度的升高细胞凋亡率逐渐升高(P<0.05);使用免疫组化光密度定量检测不同浓度的奈达铂对CNE-2细胞P53、Bcl-2、caspase蛋白表达水平的影响,可见随着奈达铂浓度的升高P53、Bcl-2蛋白表达水平逐渐降低(P<0.05),caspase蛋白表达水平逐渐升高(P<0.05).结论 奈达铂能够明显抑制鼻咽癌CNE-2细胞增殖和促进凋亡,其可能与抑制P53、Bcl-2蛋白表达和促进caspase蛋白表达有关.     

二氧化氯作为消毒剂在医院的应用

本实验证明 ,二氧化氯用于医院内消毒使用浓度及作用时间为 :一般容器用 10 0μg/ ml溶液作用 5min或 50 μg/ ml溶液作用 2 0 min;物品表面擦拭用 2 0 0 μg/ ml溶液作用 10 min;普通病房空气喷雾消毒用 2 0 0 μg/ ml溶液 8ml/ m3,作用 30 min;医疗器械浸泡用 10 0μg/ ml溶液作用 10 min以上或 2 0 0μg/ ml作用 5min以上。     

左氟沙星、氯霉素、红霉素的体外抗菌效应比较

目的　为研制开发更好的外用抗感染制剂提供科学的参考依据。方法　检测了 6种 186株常见临床感染菌对左氟沙星、氯霉素、红霉素的敏感性。结果　左氟沙星、氯霉素、红霉素对 6种临床分离细菌MIC范围为 0 .0 6 2 5～ 8.0 μg/ml,0 .12 5～ 12 8μg/ml,0 .12 5～ 2 5 6 μg/ml;MIC50 为 0 .5 2 μg/ml,2 6 μg/ml,98μg/ml,MIC90 为 5 .6 μg/ml,10 8μg/ml,2 0 8μg/ml。 结论　左氟沙星的体外抗菌活性较氯霉素、红霉素强 ,氯霉素的体外抗菌活性较红霉素强     

黄芪甲苷Ⅳ联合紫杉醇通过STAT3-NF-κB途径对胃癌细胞的作用研究

目的探讨黄芪甲苷Ⅳ联合紫杉醇对胃癌细胞的作用及其可能的机制。方法黄芪甲苷Ⅳ和紫杉醇按给药剂量分为0μg/ml组、5μg/ml组、10μg/ml组、20μg/ml组、40μg/ml组、80μg/ml组和160μg/ml组处理胃癌细胞AGS。CCK-8试剂盒检测细胞活力;集落形成实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测Bax、Bcl-2和STAT3-NF-κB途径相关蛋白的表达;Transwell实验检测细胞迁移和侵袭。结果黄芪甲苷Ⅳ在20μg/ml、40μg/ml、80μg/ml和160μg/ml浓度下可明显抑制AGS细胞活力,黄芪甲苷Ⅳ IC_(50)=63.99μg/ml;紫杉醇在10μg/ml、20μg/ml、40μg/ml、80μg/ml和160μg/ml浓度下可明显抑制AGS细胞活力,紫杉醇IC_(50)=40.08μg/ml。与对照组相比,黄芪甲苷Ⅳ组、紫杉醇组和联合组的细胞活力、克隆形成数、细胞迁移和侵袭数明显降低(P<0.05),细胞凋亡率、Bax蛋白表达明显升高(P<0.05),Bcl-2、pSTAT3/STAT3和pNF-κB/NF-κB蛋白表达明显降低(P<0.05);与联合组比较,黄芪甲苷Ⅳ组和紫杉醇组的细胞活力、克隆形成数、细胞迁移和侵袭数明显升高(P<0.05),细胞凋亡率、Bax蛋白表达明显降低(P<0.05),Bcl-2、pSTAT3/STAT3和pNF-κB/NF-κB蛋白表达明显升高(P<0.05)。结论黄芪甲苷Ⅳ联合紫杉醇可抑制胃癌细胞增殖、迁移和侵袭,促进细胞凋亡,该作用可能是通过抑制STAT3-NF-κB途径实现的。     

克拉霉素对铜绿假单胞菌生物被膜的影响

目的 :研究克拉霉素对铜绿假单胞菌生物被膜生物特性的影响。方法 :采用结晶紫染色法测定生物被膜中细菌的粘附性 ;浓度梯度渗透法测定生物被膜的渗透性 :采用卡唑 /乙醇法测定克拉霉素对藻酸盐的合成抑制作用。结果 :采用平板法 7d可建立稳定的生物被膜 ,1/4MIC克拉霉素与 8×MIC加替沙星联合应用后光密度值由 ( 0 .12 6± 0 .0 11)显著的降低至 ( 0 .114± 0 .0 12 ) ,4μg/ml组、8μg/ml组、16 μg/ml组加替沙星的渗透浓度 ( μg/ml)可由 ( 1.2 10± 0 .0 91)、( 2 .911± 0 .112 )、( 5 .911± 0 .2 13)显著的增加至 ( 1.730± 0 .132 )、( 3.911±0 .111)、( 7.90 2± 0 .35 4) ;80 μg/ml、40 μg/ml、2 0 μg/ml、10 μg/ml克拉霉素组生物被膜中藻酸盐的含量 ( μg/108CFU)由 ( 10 .0 7± 0 .5 5 )显著的降低至 ( 2 .34± 0 .2 1)、( 4 .91± 0 .16 )、( 7.2 2± 0 .36 )、( 8.82± 0 .5 0 )。结论 :克拉霉素可降低铜绿假单胞菌生物被膜中细菌的粘附性 ,增强加替沙星在生物被膜中的渗透能力 ,抑制铜绿假单胞菌藻酸盐的合成。     

枸杞多糖对β-TC6细胞胰岛素分泌及相关基因的影响

目的研究枸杞多糖(LBP)对小鼠胰岛β-TC6细胞胰岛素分泌及相关基因的影响。方法用浓度为0、12.5、25、50、100、200、300μg/ml的LBP处理β-TC6细胞,MTT法检测细胞增殖活性,ELISA法检测LBP对葡萄糖刺激胰岛素的分泌水平,RT-PCR法检测胰岛素及相关基因mRNA表达。结果 LBP呈浓度依赖性地刺激β-TC6细胞生长。在葡萄糖浓度为20mmol/L的高糖环境下,LBP≥25μg/ml时胰岛素分泌明显增多(P<0.01)。LBP 12.5μg/ml、100μg/ml能显著降低胰岛素mRNA表达(P<0.01),但对胰十二指肠同源盒因子1、葡萄糖激酶和葡萄糖转运蛋白2基因mRNA表达无明显影响。结论 LBP的促胰岛素分泌作用可能与促进β-TC6细胞增殖有关,而与胰岛素相关基因表达无关。     

黄芪甲苷IV联合紫杉醇通过STAT3-NF-κB途径对胃癌细胞的作用研究

目的 探讨黄芪甲苷IV联合紫杉醇对胃癌细胞的作用及其可能的机制.方法 黄芪甲苷IV和紫杉醇按给药剂量分为0μg/ml组、5μg/ml组、10μg/ml组、20μg/ml组、40μg/ml组、80μg/ml组和160μg/ml组处理胃癌细胞AGS.CCK-8试剂盒检测细胞活力;集落形成实验检测细胞增殖;流式细胞术检测细胞...     

紫草素对人宫颈癌HeLa细胞增殖抑制与诱导凋亡的作用研究

目的:研究紫草素对人宫颈癌HeLa细胞增殖抑制与诱导凋亡的作用。方法:10μg/ml紫草素作用于人宫颈癌HeLa细胞,观察细胞的形态学变化;MTT法测定细胞活性,检测紫草素对HeLa细胞的增殖抑制作用;分别测定半胱氨酸天冬氨酸蛋白酶(Caspase)3、8、9的活性,探究细胞凋亡的途径;流式细胞仪测定细胞凋亡率和细胞周期。结果:10μg/ml紫草素可使HeLa细胞部分死亡;1~20μg/ml紫草素可抑制人宫颈癌HeLa细胞的增殖(P<0.01或P<0.05);1、5、10、15、20μg/ml紫草素可增强Caspase-3、8、9的活性(P<0.01或P<0.05);1、5、10、15、20μg/ml紫草素在诱导人宫颈癌HeLa细胞凋亡的同时,阻断了细胞周期的进程,使S期细胞增多。结论:紫草素可抑制HeLa细胞的增殖,诱导细胞凋亡。     

姜黄素抑制人神经胶质瘤U251细胞增殖的机制

目的 探讨姜黄素对人神经胶质瘤U251细胞增殖的抑制作用及其相关机制.方法 应用0、10、20、40、60、80 μmol/L的姜黄素处理人神经胶质瘤U251细胞,检测细胞增殖、细胞周期、细胞凋亡及葡萄糖转运蛋白-3(GLUT-3)表达情况.结果 10、20、40、60、80 μmol/L姜黄素作用于U251细胞吸光度(OD)值低于0μmol/L组(P＜0.05).姜黄素组作用72 h时人神经胶质瘤U251细胞G1期细胞比例少于对照组,G2/M期细胞比例多于对照组(P＜0.05).两组细胞凋亡率比较差异无统计学意义(P＞0.05).姜黄素组GLUT-3表达水平低于对照组(P＜0.05).结论 姜黄素可以抑制人神经胶质瘤U251细胞增殖,其作用可能是通过下调GLUT-3的表达水平来实现的.     

全反式维甲酸联合猪胆酸钠对人卵巢癌细胞生长抑制作用的影响

目的 探讨维甲酸（ATRA）及猪胆酸钠（SBANa)联合反应对人卵巢癌细胞生长的抑制作用。方法 实验共分为三组：单用ATRA组、单用SBANa组及ATRA＋SBANa联合用药组。应用MTT比色法、流式细胞仪对COC1、COC2、CAOV3三种卵巢癌细胞系在不同浓度的ATRA及SBANa下进行检测。结果 ATRA及SBANa联合应用后，能增强抑制卵巢癌细胞生长作用，当达到ATRA10μmol/L SBANa 100μg/ml浓度时，G0、G1期细胞比例增加，G2M、S期细胞比例下降，细胞增殖速度较单独用药组减缓。MTT检测提示在联合用药浓度达到ATRA 10μmol/L SBANa 100μg/ml时，癌细胞抑制率最佳。结论 ATRA及SBANa联合应用可增强对卵巢癌细胞增殖的抑制作用，并在ATRA 30μmol/L SBANa 100μg/ml浓度时，促进癌细胞凋亡。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.