丹参粉针剂对自发性高血压大鼠心肌细胞凋亡的影响

目的探讨丹参粉针剂对自发性高血压大鼠(SHR)心肌细胞凋亡的作用。方法18只引种繁殖的雄性SHR随机分成3组,每组6只对照组育至第8周处死;丹参组从第8周给药,丹参粉针剂每天1 g·kg-1腹腔注射,第18周处死;高血压组用相同容量蒸馏水替代丹参针腹腔注射,余同丹参组。测量大鼠尾动脉收缩压(SBP)及左心室重量指数(LV-MI) ,采用TdT介导的原位末端缺口标记法(TUNEL)检测心肌细胞凋亡,用S-P免疫组化法检测心肌细胞Bcl-2基因蛋白表达。结果与对照组比较,高血压组SBP、LVMI、心肌细胞凋亡指数(AI)显著增加,Bcl-2蛋白阳性表达指数(PEI)降低;与高血压组比较,丹参组LVMI、AI显著降低,PEI增高,差异均有统计学意义。结论长期应用丹参粉针剂治疗,能显著上调SHR心肌细胞Bcl-2基因蛋白表达,抑制心肌细胞凋亡,预防高血压左室肥厚的形成。     

淫羊藿次苷Ⅱ改善自发性高血压大鼠左心室心肌细胞凋亡

目的观察淫羊藿次苷Ⅱ(icarisideⅡ,ICSⅡ)对自发性高血压大鼠(spontaneously hypertensive rats,SHR)心肌细胞凋亡的干预作用,并探讨其可能的机制。方法 30只13周龄♂SHR随机分为模型组、ICSⅡ低、中、高剂量组及阳性药组(n=6);同时,同源♂京都大鼠(WKY)作为对照组(n=6)。所有大鼠适应性饲养1周后,ICSⅡ低、中、高剂量组分别给予ICSⅡ4、8、16 mg·kg~(-1)(ig,qd),阳性药组给予氯沙坦20 mg·kg~(-1)至26周龄,WKY和SHR组给予等体积双蒸水。给药12周后,测量大鼠血压,处死大鼠并分离左心室,计算左心质量指数;HE染色观察左心室病理变化;TUNEL染色观察左心室心肌细胞凋亡情况;real time RTPCR检测左心室Bcl-2、Bax mRNA水平;Western blot检测左心室Bcl-2、Bax和cleaved-caspase-3蛋白表达。结果相较于WKY组,SHR组大鼠血压升高,左心质量指数增加(P<0.05);心肌细胞排列紊乱、细胞肥大明显并伴有肌丝断裂;左室心肌细胞凋亡明显,Bax mRNA水平和蛋白表达上调,Bcl-2 mRNA水平和蛋白表达下调,Bax/Bcl-2比值升高,cleaved-caspase-3蛋白表达上调(P<0.05)。与SHR组相比,ICSⅡ中、高剂量组和阳性药组血压和左心质量指数下降(P<0.05);心肌细胞排列趋于整齐、细胞肥大及心肌细胞凋亡情况均得以改善;Bax mRNA水平和蛋白表达下调,而Bcl-2 mRNA水平和蛋白表达上调,Bax/Bcl-2比值降低,cleaved-caspase-3蛋白下调(P<0.05)。结论 ICSⅡ可减轻SHR左心室心肌细胞凋亡,其机制与其降低血压和抑制线粒体凋亡途径相关。     

Caspase抑制剂对大鼠缺血再灌注心肌细胞凋亡及Bcl-2、Bax蛋白表达的影响

目的 探讨Caspase抑制剂z-VAD-fmk对缺血-再灌注(I-R)心肌细胞凋亡及Bcl-2、Bax蛋白表达的影响.方法 24只大鼠随机分为假手术(C)组、I-R对照(B)组和z-VAD-fmk治疗(A)组,以穿线结扎或松扎左冠状动脉制备大鼠心肌I-R模型,TUNEL法检测心肌细胞凋亡指数(AI),免疫组化法检测Bcl-2、Pax蛋白表达,并分析心肌组织病理学损害程度.结果 B组和A组Bcl-2、Bax基因蛋白表达水平均明显高于C组(P<0.05).A组凋亡指数显著低于B组;与B组比较,A组Bax蛋白表达水平有所下降,而Bcl-2蛋白表达水平显著升高(P<0.05),Bcl-2/Bax比值较对照组明显增加.结论 Caspase抑制剂可抑制I-R后心肌细胞凋亡,其机制可能与上调Bcl-2和下调Bax蛋白表达有关.     

术前腹腔动脉灌注化疗对中晚期胃癌细胞凋亡和增殖的影响

目的 探讨术前腹腔动脉灌注化疗对中晚期胃癌细胞凋亡和增殖的影响。方法 采用切口末端标记法(TUNEL)、增殖细胞核抗原(PC-NA)和凋亡抑制蛋白(Bcl-2)免疫组化检测技术,观察比较术前腹腔动脉灌注化疗组31例和未化疗手术组32例中晚期胃癌标本的凋亡指数、增殖指数和BCcL-2蛋白表达强度的差异。结果 术前化疗组凋亡指数明显高于对照组(P<0.01),增殖指数明显降低(P<0.01),Bcl-2蛋白表达强度明显低于对照组(P<0.01)。凋亡指数与Bcl-2分值是显著负相关(P<0.01),凋亡指数与增殖指数呈中度正相关(P<0.05)。治疗组治愈性切除率明显高于对照组(P<0.01),3年生存率高于对照组(P<0.05)。结论 术前腹腔动脉插管化疗可诱导中晚期胃癌细胞凋亡,降低肿瘤细胞的增殖活力,提高治愈性切除率,改善患者生存率。诱导胃癌细胞凋亡可能与化疗药物降低Bcl-2基因蛋白的表达有关。     

硫辛酸对多柔比星致大鼠心肌细胞损伤凋亡的保护作用研究

目的:研究硫辛酸对多柔比星致大鼠心肌细胞损伤凋亡的保护作用。方法:将大鼠随机分为正常对照组、模型组和低、中、高剂量[10、20、40mg(/kg·d)]硫辛酸组,每组10只,后4组腹腔注射多柔比星3mg/kg,隔日1次,连用6次建模。硫辛酸组于注射多柔比星第2天起腹腔注射相应浓度的硫辛酸,每日1次,连用10d。多柔比星末次给药24h后处死大鼠,观察各组大鼠心肌结构变化,检测心肌细胞凋亡情况和心肌细胞中抑制细胞凋亡因子Bcl-2、凋亡基因Bax蛋白表达情况。结果:与正常对照组比较,模型组大鼠肌丝排列疏松、紊乱,大部分线粒体形成空泡,其心肌细胞凋亡指数和Bax蛋白表达均明显增强,Bcl-2蛋白表达明显降低(P<0.01);与模型组比较,低、中、高剂量硫辛酸组心肌细胞病理形态学变化均改善,且凋亡指数和Bax蛋白表达均明显降低,Bcl-2蛋白表达明显增强(P<0.01)并呈剂量依赖性。结论:硫辛酸可拮抗多柔比星所致大鼠心肌细胞损伤,其作用机制可能与抑制Bax蛋白表达、增强Bcl-2蛋白表达有关。     

卡维地洛、缬沙坦及其合用对自发性高血压大鼠心肌细胞凋亡、P53及Bcl-2蛋白表达的影响

目的探讨卡维地洛(Carvedilol,Car)、缬沙坦(Valsartan,Val)及其合用对自发性高血压大鼠(SHR)心肌细胞凋亡、心肌Bcl-2、P53蛋白表达的影响。方法6周龄雄性SHR24只,随机分为4组(n=6):一组为对照组,其他3组为干预组,分别予NS5ml、Cal(20mg·kg-1·d-1)、Val(40mg·kg-1·d-1)和Car+Val合用(剂量同上)干预8周后,予2%戊巴比妥钠40mg/kgIP麻醉、开胸取心脏,取左心室游离壁心肌于10%中性福尔马林中固定24小时,继之脱水,石蜡包埋及切片,采用TUNEL法、免疫组化法分别检测心肌细胞凋亡率(cardiacmyocyteapoptosisrate,CMAR),Bcl-2及P53蛋白表达率。结果与对照组比较,各干预组CMAR及P53蛋白表达率均明显降低,而Bcl-2蛋白表达率却明显增高,尤以合用组更明显(P<0.01)。与Car组、Val组比较,合用组CMAR及P53蛋白表达率均降低,而Bcl-2蛋白表达率均增高(P<0.01)。与Car组比较,Val组CMAR、Bcl-2及P53蛋白表达率,差异无显著性意义(P>0.05)。各组中P53蛋白表达率与CMAR呈正相关;而Bcl-2与CMAR呈负相关((P<0.05)。结论CarVal均可抑制SHR心肌细胞凋亡,二者合用效应更明显,可能与两药均能上调BCL-2蛋白表达和下调P53蛋白表达有关。     

多肽药物汇利心康对慢性心衰模型大鼠心肌细胞凋亡的抑制作用研究

目的:研究多肽药物汇利心康(HLXK)对慢性心衰模型大鼠心肌细胞凋亡的影响。方法:取大鼠随机分为正常对照组、模型组、阳性对照(灌胃氯沙坦钾6mg·kg-1,每日1次)组和HLXK低、中、高剂量(腹腔注射10、30、90μg·kg-1,每日2次)组,每组8只,后5组腹腔注射盐酸多柔比星(隔天给药,15d)建立慢性心衰模型,同时给予相应药物。10周后用高频彩超测定各组大鼠左心室收缩末期容积(LVESV)和射血分数(EF),免疫组化法检测心肌组织中Bcl-2、Bax蛋白表达。结果:与正常对照组比较,模型组大鼠LVESV明显增加,EF明显减少(P<0·01),表明其心功能明显减退;且Bcl-2蛋白表达明显减弱,Bax蛋白表达明显增强(P<0·01)。与模型组比较,阳性对照组和HLXK中、高剂量组LVESV明显减少,EF明显增加,Bax蛋白表达明显减弱,Bcl-2蛋白表达明显增强(P<0·05或P<0·01);HLXK低剂量组仅Bax蛋白表达明显减弱,Bcl-2蛋白表达明显增强(P<0·05)。结论:HLXK可能通过增强模型大鼠Bcl-2蛋白表达、抑制Bax蛋白表达,从而抑制心肌细胞凋亡。     

延黄消心痛胶囊对实验性急性心肌梗死大鼠Bcl-2、Bax蛋白表达及心肌细胞凋亡的影响

目的观察延黄消心痛胶囊对实验性急性心肌梗死大鼠Bax、Bcl-2蛋白及心肌细胞凋亡的影响,探讨其干预心肌梗死的机制。方法 60只SD大鼠随机分为假手术组、模型组、延黄消心痛胶囊高、中、低剂量组和地奥心血康对照组,药物干预7d后,结扎SD大鼠结扎左冠状动脉前降支建立心梗动物模型。应用免疫组化法观察Bax、Bcl-2蛋白表达;应用TUNEL法观察大鼠心肌缺血区的心肌细胞凋亡情况。结果①模型组的Bax蛋白阳性表达显著高于假手术组(P值均<0.05),Bcl-2蛋白表达显著下降(P<0.01);干预药高、中、低剂量组和对照组的Bax蛋白阳性表达显著低于模型组(P<0.01),Bcl-2蛋白表达显著上调(P<0.01)。②模型组与假手术组心肌细胞凋亡指数的差异有统计学意义(P<0.01),干预药高、中、低剂量组和对照组均与假手术组、模型组的差异有统计学意义(P值均<0.01)。结论延黄消心痛胶囊能够抑制心肌梗死后大鼠心肌细胞凋亡,其机制与Bcl-2蛋白表达上调、Bax蛋白表达下调有关。     

丹参对CAPD大鼠腹膜间皮细胞凋亡及凋亡相关基因的影响

目的:探讨丹参对持续性非卧床式腹膜透析(CAPD)相关性腹膜硬化的治疗价值。方法:将40只大鼠随机分为对照组(A组,给予0.9%NaCl40mL/d),凋亡模型组(B组,给予4.25%腹透液40mL/d),低剂量丹参组(C组,给予4.25%腹透液40mL/d和600mg·kg-·1d-1丹参),高剂量丹参组(D组,给予4.25%腹透液40mL/d和2400mg·kg-·1d-1丹参)。8周后检测腹膜间皮细胞凋亡及促凋亡基因(Fas)和抑凋亡基因(Bcl-2)的蛋白表达,计算凋亡细胞指数(AI)和基因的蛋白阳性表达指数(PEI)。结果:B组FasPEI高于A组(P<0.01),C、D组FasPEI明显低于B组(P<0.01),与A组相近(P>0.05)。A、B、C组Bcl-2PEI差别无统计学意义,D组Bcl-2PEI明显高于A、B组(P<0.01)。B组AI明显高于A组,C、D组AI明显低于B组(P<0.01)。结论:丹参可通过下调Fas基因的蛋白表达,上调Bcl-2基因的蛋白表达,对CAPD时腹膜间皮细胞凋亡产生抑制作用。     

心肌挫伤后心肌细胞凋亡的实验研究

目的探讨心肌挫伤对家兔心肌细胞凋亡及B细胞淋巴瘤(Bcl-2)和白血病-2相关X基因(Bax)蛋白表达的影响。方法健康家兔40只,随机分为3组,参考组(n=20)、对照组(n=10)、心肌挫伤组(n=10)。利用BIM-Ⅱ型生物撞击机制备心肌挫伤模型,应用酶联免疫吸附剂测定(ELISA)法测定心肌肌钙蛋白I(cTnI),应用原位末端标记法(TUNEL)法并结合流式细胞术观察心肌细胞凋亡发生的情况,应用免疫组化SABC法观察Bcl-2和Bax蛋白的表达。对照组除不撞击胸部外,余处理均同MC组。结果参考组与MC组在伤后4h血清cTnI含量均高于各自伤前及对照组相应时相点(P<0.05);对照组未见TUNEL阳性细胞,凋亡率也为零,MC组心肌细胞凋亡指数(AI)和凋亡率(AR)分别为(15.6860±0.6887)%和(6.9320±0.6380)%,与对照组比较,均显著增高(P<0.05);与对照组比较,MC组的Bcl-2蛋白和Bax蛋白表达光密度值均明显增高(P<0.05)。结论心肌挫伤可诱导家兔心肌细胞的凋亡,且与Bcl-2和Bax这一对凋亡调控基因的表达变化有着某种联系。     

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.     

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.     

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.     

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.     

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.