穿膜融合多肽TAT N24对Jurkat细胞增殖的影响

摘要目的观察穿膜融合多肽TAT N24对急性T细胞白血病Jurkat细胞增殖的影响。方法以穿膜融合多肽TAT N24处理Jurkat细胞,噻唑蓝（MTT）法观察细胞的生长情况,应用流式细胞仪检测Jurkat细胞周期进程。结果穿膜融合多肽TAT N24处理急性T细胞白血病Jurkat细胞48 h,细胞生长受到抑制,随着多肽浓度的增加,抑制作用增强,表现出明显的浓度依赖性（P＜0.05）;48 h后细胞生长明显受抑,并随时间延长生长受抑更加明显,表现出时间依赖性（P＜0.05）。空白对照组Jurkat细胞中G0/G1期细胞数为（41.91±1.96）%,S期和G2/M期细胞数分别为（52.16±1.76）%和（5.93±0.30）%;对照多肽组Jurkat细胞中G0/G1期细胞数为（46.38±2.31）%,S期和G2/M期细胞数分别为（44.22±1.69）%和（9.40±0.98）%;TAT N24组Jurkat细胞中G0/G1期细胞数为（54.83±1.92）%,S期和G2/M期细胞数分别为（30.75±2.27）%和（14.42±0.68）%。结论融合多肽TAT N24可以有效抑制急性T细胞白血病Jurkat细胞的增殖,阻滞其周期进程。     

一氧化氮抑制常氧和低氧培养的肺动脉内皮细胞增殖

探讨一氧化氮(NO)在常氧和无氧(0% O₂)条件下对肺动脉内皮细胞(PAEC)增殖的影响. 结果表明，无氧和NO合酶抑制剂L-硝基精氨酸(L-NA 2.5 mmol·L^-1)培养24 h对PAEC形态无明显影响，NO供体SIN-1(0.1-1.0 mmol·L^-1)使常氧与无氧条件下培养的PAEC贴壁细胞明显减少；与常氧组相比，无氧培养24 h使PAEC的［³H］TdR参入降低53%；L-NA对PAEC的［³H］TdR参入无明显影响，SIN-1则浓度依赖性地抑制PAEC的［³H］TdR参入. 流式细胞分析表明，无氧使进入S期和G₂/M期的细胞分别增加22%和144%，而进入G₀/G₁期的细胞降低49%(P<0.01)；SIN-1(0.5 mmol·L^-1)具有与低氧相似的作用，使进入S期和G₂/M期的细胞分别增加42%和104%，而进入G₀/G₁期的细胞减少41%(P<0.01). 无氧加入SIN-1后使S期细胞增加更为显著. 结果说明低氧和外源性NO均抑制PAEC的增殖，其抑制作用环节可能是阻止G₂/M期细胞向G₀/G₁期过渡.     

槲皮素对HL-60细胞周期的影响

本文报道了槲皮素对人早幼粒白血病细胞株HL-60细胞的增殖和细胞周期的影响. 结果表明槲皮素能抑制HL-60细胞的增殖， 呈剂量依赖关系， 当槲皮素作用24， 48， 72 h后， 其IC₅₀分别为 46.6, 28.7, 14.9 μmol·L^-1流式细胞仪分析表明，槲皮素能明显增加HL-60的G₂-M期细胞，而相对减少G₀/G₁细胞之百分比，且呈剂量依赖关系，当去除槲皮素时，该作用是可逆的. 结果表明槲皮素对肿瘤细胞的生长抑制作用可能与其对细胞周期的影响有关.     

1-(2,6-二甲基苯氧基)-2-(3,4-二甲氧基苯乙氨基)丙烷盐酸盐对内皮素促血管平滑肌细胞增殖及癌基因表达的影响

采用内皮素-1(ET-1 0.1 μmol·L^-1)建立培养的血管平滑肌细胞增殖模型，用［³H］胸腺嘧啶核苷(［³H］TdR)参入法, 流式细胞术, 免疫细胞化学及Northern blot方法, 观察了1-(2,6-二甲基苯氧基)-2-(3,4-二甲氧基苯乙氨基)丙烷盐酸盐(DDPH 0.1 μmol·L^-1)对血管平滑肌细胞增殖的作用及对原癌基因及抑癌基因的影响. 结果发现: DDPH能逆转ET-1所致［³H］TdR参入量增多, 阻止血管平滑肌细胞由静止期 (G₀/G₁期)进入DNA合成期(S期)和有丝分裂期(G₂/M期), 并能逆转ET-1引起的c-fos, c-myc, c-sis原癌基因相关抗原及 mRNA表达增强, P53抑癌基因相关抗原及mRNA表达减弱. 提示DDPH能抑制血管平滑肌细胞增殖, 与癌基因调控的分子生物学机理有关.     

黄芩苷通过硫氧还原蛋白介导对SH-SY5Y细胞保护作用

目的 研究黄芩苷对过氧化氢(H₂O₂)损伤后的成人神经细胞瘤细胞母细胞SH-SY5Y细胞的影响,进一步探讨黄芩苷保护SH-SY5Y细胞的作用机制. 方法 不同浓度黄芩苷预先处理正常培养的SH-SY5Y细胞24 h,200 μmol.L^-1 H₂O₂损伤上述SH-SY5Y细胞24 h,采用实时荧光定量聚合酶链反应(PCR)技术检测各实验组细胞的硫氧还原蛋白(Trx)mRNA的表达;采用酶联免疫吸附测定(ELISA)技术检测各实验组细胞的Trx 蛋白的表达. 结果 黄芩苷在50～300 μmol.L^-1浓度范围内对SH-SY5Y细胞无细胞毒作用,0,50,100和200 μmol.L^-1黄芩苷组SH-SY5Y细胞存活率分别为(0.410±0.096),(1.304±0.101),(0.899±0.089)和(0.609±0.023),50,100和200 μmol.L^-1均明显高于H₂O₂ 损伤组(0.339±0.073)的存活率,正常组,H2O2损伤组,50 ,100和200 μmol.L^-1黄芩苷组Trx mRNA的表达水平分别为(1.023±0.014),(0.021±0.004),(0.054±0.005),(0.071±0.013)和(0.042±0.004);正常组Trx蛋白表达水平为(26.230±0.857),H₂O₂损伤组为(19.230±0.982),50,100和200 μmol.L^-1黄芩苷组Trx蛋白表达水平分别为(22.440±0.888),(23.020±1.070),(22.330±1.067). 结论 黄芩苷对H₂O₂损伤后的SH-SY5Y细胞有保护作用,其作用机制可能与黄芩苷抑制H2O2诱导的Trx表达下调作用有关,起到抗氧化应激及抗细胞凋亡的作用.     

黄芩苷对SH-SY5Y细胞损伤的Bcl-2和Bcl-xL mRNA基因表达的影响

目的 探讨黄芩苷对过氧化氢(H₂O₂)诱导的SH-SY5Y细胞损伤的Bcl-2和Bcl-xL mRNA表达的影响. 方法 建立人神经母细胞瘤SH-SY5Y细胞的体外H₂O₂损伤模型,采用噻唑蓝(MTT)法检测不同浓度黄芩苷对SH-SY5Y细胞存活率的影响,采用Real-time PCR法检测各组细胞的Bcl-2和Bcl-xL表达水平的变化. 结果 50,100,200 μmol.L^-1黄芩苷组细胞存活率分别为130.4%,89.9%和60.9%,H₂O₂损伤组细胞存活率为33.9%,50,100 μmol.L^-1黄芩苷组与H₂O₂损伤组比较,差异有统计学意义(P<0.01). 50,100,200 μmol.L^-1黄芩苷组Bcl-2 mRNA表达量分别为(11.48±0.48),(7.37±1.57),(7.39±2.01),H₂O₂损伤组为(5.84±0.58);50,100,200 μmol.L-1黄芩苷组Bcl-xL mRNA表达量分别为(19.96±2.22),(11.36±3.94),(13.07±2.37),H₂O₂损伤组为(7.95±0.58),50 μmol.L^-1组Bcl-2 和Bcl-xL mRNA与H^₂O^₂损伤组比较,差异均有统计学意义(均P<0.05). 结论 黄芩苷对H₂O₂诱导的SH-SY5Y细胞损伤的保护作用,可能与黄芩苷上调Bcl-2和Bcl-xL的表达而起到抗凋亡的作用有关.     

非那雄胺胶囊的人体生物等效性研究

目的 研究国产非那雄胺胶囊与进口非那雄胺片(保列治)的人体生物等效性。方法 采用LC-MS法测定18例健康男性单次交叉口服试验制剂及对照制剂各5 mg后血浆中不同时间点的浓度,经SPSS软件统计拟合,计算其药动学参数和相对生物利用度,评价两种制剂的生物等效性。结果非那雄胺胶囊试验制剂和对照制剂的AUC0→24分别为（355.7±63.7）和(363.1±65.6 ) ng·h·mL^-1, C_max分别为(51.2±7.4)和（56.3±5.5 ） ng·mL^-1,t_max分别为（2.1±0.9）和（1.9±0.4） h。试验制剂相对于对照制剂的人体生物利用度为（98.6±11.8）%,AUC_0→24,Cmax经对数转换后,经双单侧t检验并计算AUC_0→24,C_max的90%可信区间分别在80%～125%,70%～143%。结论 实验表明两种制剂具有生物等效性.     

多剂量口服硝苯地平控释片的人体药动学及生物等效性研究

目的 以聚维酮/共聚维酮替代聚氧乙烯作为促渗透聚合物制备新型渗透泵型硝苯地平控释片,研究多剂量口服硝苯地平控释片的人体药动学特点,计算相对生物利用度,并评价该制剂的生物等效性。方法 入选的24名男性健康受试者随机交叉给药,分别每日口服30 mg试验制剂或参比制剂,剂量均为30 mg·次^-1·d^-1,连续服药7 d。采用LC-MS/MS法测定血药浓度,用DAS软件计算药代动力学参数及相对生物利用度,并求证硝苯地平控释片试验制剂与参比制剂的生物等效性。结果 连续给予30 mg试验制剂和参比制剂后获得主要药代动力学参数如下：T_1/2β分别为(9.0±2.2)h和(9.5±3.8)h;T_max分别为(8.1±7.4)h和(6.7±4.1)h;C_ssmax分别为(52.7±28.2)μg·L^-1和(44.5±22.6)μg·L^-1;C_ssmin分别为(29.4±22.2)μg·L^-1和(28.7±15.8)μg·L^-1;AUC_0-60分别为(1 087.4±671.6)μg·L^-1·h和(1 040.2±518.4)μg·L^-1·h;AUC_0-∞分别为(1 113.1±677.2)μg·L^-1·h和(1 074.5±519.8)μg·L^-1·h,AUCSS分别为(809.1±454.0)μg·L^-1·h和(713.9±382.2)μg·L^-1·h。经计算试验制剂对于参比制剂的平均相对生物利用度F值：FAUC_ss为(115.2±29.2)%,FAUC_0-60为(103.4±30.2)%,FAUC_0-∞为(102.2±29.7)%。波动系数DF分别为(77.8±52.1)%和(55.5±30.5)%。两种制剂的C_max、AUC_0-60、AUC_0-∞及AUC_SS经对数转换后90%置信区间的计算结果为：C_max(100.9~126.0)%,AUC_0-60 (87.5~111.8)%,AUC_0-∞ (86.8~110.3)%,AUC_SS为(98.8~124.4)%,T_max 经非参数秩和检验无显著性差异。整个试验期间,受试者均未出现药物不良反应。结论 两种硝苯地平控释片为生物等效制剂。聚维酮/共聚维酮可以替代聚氧乙烯作为渗透泵型控释制剂的主要功能性辅料。     

纳米锰锌铁氧体颗粒对L-02细胞的氧化损伤作用

目的 探讨磁性锰锌铁氧体纳米颗粒(Mn_0.5Zn_0.5Fe₂O₄)对人肝细胞株L-02的毒性作用机制。方法 Mn_0.5Zn_0.5Fe₂O₄ 800 mg·L^-1作用L-02细胞48 h,透射电镜观察细胞形态及超微结构的变化。Mn_0.5Zn_0.5Fe₂O₄ 200, 400和800 mg·L^-1作用48 h后,检测L-02细胞内丙二醛(MDA)的含量、超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)的活性;荧光染色观察凋亡细胞形态;流式细胞术检测细胞周期及凋亡;荧光定量PCR仪检测胱天蛋白酶3 mRNA表达。结果 Mn_0.5Zn_0.5Fe₂O₄ 800 mg·L^-1作用48 h后,纳米颗粒进入细胞内,细胞膜发生破损,细胞器消失,染色体异常聚集。与正常对照组比较,Mn_0.5Zn_0.5Fe₂O₄ 200~800 mg·L^-1使细胞内MDA含量逐渐升高,SOD与GSH活性逐渐降低(P<0.05)。Mn_0.5Zn_0.5Fe₂O₄可使细胞周期发生改变,G₀/G₁期细胞百分率有降低的趋势,S期和G₂/M期细胞百分率有升高的趋势。Hoechst33258显示明显的细胞凋亡形态。Mn_0.5Zn_0.5Fe₂O₄可引起L-02细胞发生剂量依赖性的细胞凋亡,Mn_0.5Zn_0.5Fe₂O₄ 800 mg·L^-1作用48 h后,细胞凋亡率达到30.3%,是对照组细胞凋亡率(2.4%)的12.6倍。胱天蛋白酶3 mRNA表达量先增加后降低,但都明显高于正常对照组(P<0.05)。结论 Mn_0.5Zn_0.5Fe₂O₄可破坏细胞膜完整性并进入细胞内,诱导细胞发生氧化应激,改变细胞周期,引发细胞凋亡,产生细胞毒性。     

国产右旋布洛芬缓释胶囊在大鼠体内的药动学及生物利用度研究

目的 考察自制和市售右旋布洛芬缓释胶囊在大鼠体内的药动学性质与生物等效性.方法建立反相高效液相色谱法,测定右旋布洛芬在血浆中的浓度,进行药动学和相对生物利用度研究.结果自制和市售右旋布洛芬缓释胶囊的主要药动学参数C_max分别为(1 173.87±281.68),(1 186.06±268.79) μg.mL^-1;AUC_(0-t)分别为(4 276.53±578.59),(4 489.83±645.73) mg.L^-1.h;AUC_(0-∞)分别为(5 095.58±683.82),(5 466.37±753.35) mg.L^-1.h;t_max分别为(1.33±0.26),(1.58±0.49) h;各参数间比较差异无统计学意义(P>0.05),以AUC_(0-∞)计算自制右旋布洛芬缓释胶囊的相对生物利用度为(93.2±12.5)%.结论自制右旋布洛芬缓释胶囊与市售制剂之间生物等效.     

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.     

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.     

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.     

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.