姜黄素抑制子宫颈癌HeLa细胞的作用

目的：探讨姜黄素抑制子宫颈癌HeLa细胞的作用及其机制。方法以细胞培养，镜下观察细胞形态学改变和计数法测定生长曲线；用3H-脱氧胸苷掺入法测定对DNA合成的影响，以MTT比色法检测给药后HeLa细胞的增殖抑制情况。结果姜黄素作用HeLa细胞后，癌细胞生长延缓并萎缩，胞质粗糙，有大量颗粒状物堆积，而且药物浓度越大，形态学改变越明显；生长曲线测定、3H-脱氧胸苷掺入法及MTT比色法实验结果显示，姜黄素对HeLa细胞的增殖和生长有显著的抑制作用并呈明显的时间一剂量依赖关系，当姜黄素浓度为20μg／ml以上时，其对HeLa细胞的掺入抑制率高于5一Fu20μg／ml对该细胞的掺入抑制率。结论姜黄素对HeLa细胞具有直接杀伤作用，其机制可能通过干扰细胞代谢，改变细胞外暌的性质抑制肿瘤细胞增值。     

姜黄素对人宫颈癌Hela细胞增殖的抑制作用

目的:研究姜黄素对人宫颈癌Hela细胞生长的抑制作用。方法:姜黄经75%乙醇提取、纯化得姜黄素。MTT法检测姜黄素对体外培养Hela细胞增殖的抑制作用;倒置显微镜观察Hela细胞形态学变化;Western blot法检测抑癌基因p53蛋白的表达;流式细胞仪检测Hela细胞凋亡和周期分布。结果:姜黄素对Hela细胞生长24 h的最佳抑制质量浓度为116.68μg/ml,抑制率为63.20%;11.67、35.00、58.34μg/ml姜黄素培养24 h,人宫颈癌Hela细胞数目明显减少,细胞变圆、缩小、老化,核质发散,并与质量浓度呈正相关。29.17、145.85μg/ml姜黄素可增强p53蛋白的表达;11.67、35.00、58.34μg/ml姜黄素可促进人宫颈癌Hela细胞凋亡,并与质量浓度呈正相关,G0/G1期细胞和S期细胞逐渐减少,G2/M期细胞逐渐增多。结论:姜黄素对人宫颈癌Hela细胞增殖有明显抑制作用,并能促进其凋亡和p53的表达,阻滞Hela细胞的G2/M期。     

藤茶双氢杨梅树皮素对人肝癌BEL-7404细胞增殖的抑制作用

目的：探讨藤茶双氢杨梅树皮素（APS）的抗肿瘤作用。方法：以MTT法、生长曲线法、克隆形成法观察APS对人肝癌BEL-7404细胞的体外抑制作用。结果：MTT法中，APS对BEL-7404细胞的IC50为22．99μg／mL；细胞生长曲线法提示其对BEL-7404细胞生长有明显抑制作用；克隆形成法中，药物浓度在9．88μg／mL以上时抑制率为100％，药物浓度为6．58μg／mL时抑制率为69．65％。结论：APS对BEL-7404细胞的增殖有明显的抑制作用。     

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。     

熊果酸的抗肿瘤作用

目的 研究熊果酸的抗肿瘤活性.方法 体外用MTT比色法测定熊果酸对肿瘤细胞HepG2和S180的增殖抑制作用,再用DNA凝胶电泳法测定熊果酸对HepG2细胞凋亡的诱导作用;体内采用小鼠实体瘤模型,观察熊果酸对HepG2和S180实体瘤的抑制作用.结果 熊果酸对HepG2和S180的增殖有良好的抑制作用,48h内对细胞半数生长的抑制剂量(IC50)分别为12.5μg/ml和13.9μg/ml.同时熊果酸能够诱导HepG2发生凋亡.20.0μg/ml熊果酸对HepG2和S180实体瘤抑制率分别达到52.98%和44.41%.结论 熊果酸通过抑制增殖和诱导凋亡对肿瘤细胞株HepG2和S180表现出良好的抗肿瘤活性.     

螺旋藻多糖对HeLa细胞生长的影响

目的；研究螺旋藻多糖（polysaccharide from Spirulina platensis,PSP)对体外培养的HeLa细胞生长的影响。方法：MTT法测定螺旋藻多糖的抗肿瘤活性；光镜观察螺旋藻多糖对HeLa细胞形态学的影响；流式细胞术检测螺旋藻多糖对肿瘤细胞周期的影响。结果：随螺旋藻多糖浓度的增加，HeLa细胞存活率逐渐降低。抑制率逐渐增加；光镜下的观察显示，螺旋藻多糖作用24-48h后，细胞出现明显的形态学改变；流式细胞术证实螺旋藻多糖对肿瘤细胞存在周期特异性，使细胞发生G1期阻滞。结论：螺旋藻多糖抑制HeLa细胞的增殖，可能与该细胞发生G1期阻滞有关。     

黄芪总苷对肺腺癌SPCA-1细胞增殖的抑制作用

目的:研究黄芪总苷对肺腺癌SPCA-1细胞增殖的抑制作用。方法:以0(空白对照)、7.8、15.6、31.2、62.5、125.0、250.0μg/ml黄芪总苷培养细胞48 h,MTT法测定细胞活力并计算半数抑制浓度(IC50);末端脱氧核苷酸转移酶介导的脱氧尿苷(d UTP)和缺口末端标记测定(TUNEL)法、流式细胞仪检测细胞凋亡情况。结果:与空白对照比较,以7.8、15.6、31.2、62.5、125.0、250.0μg/ml黄芪总苷培养细胞48 h后,细胞活力降低(P<0.01),IC50为61.75μg/ml;15.6、31.2、62.5、125.0、250.0μg/ml黄芪总苷培养细胞48 h后,细胞凋亡率升高。结论:黄芪总苷具有一定抗肿瘤作用,其机制可能与抑制癌细胞增殖、促进其凋亡有关。     

藤茶蛇葡萄素抗人胃癌细胞作用的实验研究

目的:探讨藤茶蛇葡萄素的体外抗肿瘤作用。方法:采用MTT法、生长曲线法、细胞集落形成法观察蛇葡萄素对人胃癌SGC-7901细胞的抑制作用。结果:蛇葡萄素能明显抑制体外培养的SGC-7901细胞的生长,MTT法IC50为11.19μg/mL;细胞生长曲线法提示其对SGC-7901细胞的生长也有明显的抑制作用;集落形成法,当药物浓度在9.90μg/mL以上时抑制率为100%,当药物浓度为6.60μg/mL时抑制率为72.3%,当药物浓度为4.40μg/mL时抑制率为36.0%。结论:蛇葡萄素对体外SGC-7901细胞的生长有明显的抑制作用。     

红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用

目的探讨红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用及其作用机制。方法以MTT法检测红三叶异黄酮20、40、80μg/m L分别在24、48、72 h对人宫颈癌HeLa细胞增殖作用的影响,并在倒置显微镜下观察HeLa细胞形态学改变。流式细胞仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡的影响。实时荧光定量PCR仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡中调节基因Bax、Bcl-2表达水平的影响。结果红三叶异黄酮对人宫颈癌HeLa细胞的增殖抑制作用,表现为时间、浓度的相关性。红三叶异黄酮对HeLa细胞的形态学改变,表现为随药物浓度增加细胞固缩变小,并出现凋亡小体。流式细胞仪检测分析显示红三叶异黄酮能引起HeLa细胞凋亡,并随药物浓度的增加而明显,当红三叶异黄酮达到一定有效浓度时,诱导其细胞凋亡。通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖,能使Bax表达上升、Bcl-2表达下降,且呈剂量相关性。结论红三叶异黄酮对人宫颈癌HeLa细胞的增殖具有抑制作用,并呈一定的剂量和时间相关性,提示通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖。     

JNK 信号转导通路在黄芩苷诱导肝癌细胞凋亡中的作用

目的：探讨黄芩苷对人肝癌 HepG2细胞生长的抑制作用及其对 c-Jun 氨基末端激酶(JNK)信号通路的影响。方法50、100、150μg/ ml 的黄芩苷分别与人肝癌 HepG2细胞共同培养24、48和72 h,MTT 测定细胞生长抑制率;采用 Western blotting 方法检测黄芩苷处理72 h 后 HepG2细胞 JNK 和 p-JNK 的表达变化并观察 JNK 抑制剂对细胞凋亡的影响。结果黄芩苷对人肝癌 HepG2细胞增殖抑制作用呈时间依赖性,黄芩苷浓度低于100μg/ ml 其对细胞的抑制作用随浓度升高而增强,超过100μg/ ml 其对细胞的抑制作用不再增强。在黄芩苷处理后 HepG2细胞 p-JNK 蛋白表达上调,使用 JNK 抑制剂后,能阻断 JNK 蛋白的磷酸化,并且降低黄芩苷诱导细胞凋亡的能力。结论黄芩苷通过激活 JNK 信号通路诱导人肝癌 HepG2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.     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.