紫杉醇联合抗HER2单抗对人卵巢癌细胞株A2780的作用

目的:体外观察紫杉醇及抗HER2单抗对人卵巢癌细胞株A2780增殖的抑制作用及两者联合应用时的抑制率及相互作用.方法:采用MIT法测定两者单用或合用对A2780的增殖抑制率,利用中效原理判定两药合用的效果,用流式细胞仪观察细胞凋亡及细胞周期的变化.结果:两种药单独应用时随药物剂量增加,其效应也增加,紫杉醇的中效浓度为6.98 nmol·L-,抗HER2单抗的中效浓度为8.69 μg·ml-1;两药合用时它们的中效浓度为2.92 nmol·L-1 0.49 μg·ml-.大剂量(紫杉醇＞13.02 nmol·L-,抗HER2单抗＞2.17 μg·ml-1)合用时是拮抗作用(CI＞1),小剂量(紫杉醇＜13.02 nmol·L-,抗HER2单抗＜2.17 μg·ml-1)合用时是协同作用(CI＜1).结论:紫杉醇联合抗HER2单抗对人卵巢癌细胞株A2780的生长有抑制作用,它们合用时大剂量是拮抗,而小剂量则为协同作用.     

靶向HER-2 mRNA反义寡核苷酸对SK-BR-3乳腺癌细胞Caspase-3蛋白表达的影响

目的　检测HER 2特异性的反义寡核苷酸HA82 4对SK BR 3乳腺癌细胞Caspase 3蛋白表达的影响。 方法　选用HER 2高表达的SK BR 3乳腺癌细胞株作为试验的细胞株。HA82 4合成如前所述 ,HA4为已报道的阳性序列 ,Scramble设定为随机对照序列。上述药物分别以浓度为 2 0 0nmol·L-1孵育SK BR 3细胞 8h和 36h ,之后采用逆转录PCR法与免疫细胞化学技术检测SK BR 3细胞HER 2mR NA及Caspase 3蛋白表达水平。 结果　与Scramble对照序列相比 ,HA82 4、HA4能抑制HER 2mRNA的表达 ,HA82 4作用更强 ;与Scramble相比 ,HA82 4、HA4对Caspase 3蛋白表达水平则无影响。结论　HA82 4、HA4这两种靶向HER 2mRNA反义药物对SK BR 3乳腺癌细胞株增殖的抑制作用可能与激活Caspase 3蛋白无关。     

黄癸素对三阴性乳腺癌MDA-MB-231细胞的体内外抑制作用

目的观察黄癸素对人三阴性乳腺癌(TNBC)细胞MDA-MB-231及其裸鼠模型肿瘤的抑制作用。方法采用MTT法观察黄癸素对乳腺癌细胞MCF-7及MDA-MB-231的增殖抑制作用;建立MDA-MB-231细胞裸鼠移植瘤模型,观察黄癸素灌胃给药对荷瘤的抑制作用。TUNEL法检测肿瘤组织中细胞凋亡状况。结果黄癸素可明显抑制MDA-MB-231细胞增殖,具有明显的浓度依赖性和时间依赖性,作用48h的IC50为(1.26±0.30)mg·L-1,而对MCF-7细胞的IC50则为(32.07±9.09)mg·L-1。在体实验结果显示,中、高剂量(90、120 mg·kg-1)黄癸素对MDA-MB-231细胞裸鼠移植瘤有明显的抑制作用,抑瘤率分别为42.26%和71.57%(P<0.01)。TUNEL法表明黄癸素组细胞凋亡率明显高于空白组。结论黄癸素有抑制人三阴性乳腺癌细胞MDA-MB-231肿瘤生长、促进瘤组织凋亡的作用。     

人参皂苷Rg3对乳腺癌细胞MDA-MB-453增殖、迁移及凋亡的影响

目的探讨人参皂苷Rg3对乳腺癌细胞MDA-MB-453增殖、迁移及凋亡的影响。方法实验组采用MTT法检测不同浓度人参皂苷Rg3(10、20、30、40、50、60、70和80μmol/L)对乳腺癌细胞MDA-MB-453的抑制作用;对照组不予人参皂苷Rg3干预。采用集落形成实验、细胞划痕实验和TUNEL染色分别检测两组MDA-MB-453细胞增殖、迁移及凋亡变化。流式细胞术检测两组细胞周期分布比例。Western blot检测两组凋亡相关蛋白和信号传导与转录激活因子3(STAT3)信号通路相关蛋白表达。免疫荧光技术检测STAT3的亚细胞定位。结果人参皂苷Rg3对MDA-MB-453细胞增殖有浓度依赖性抑制作用(P<0.05)。人参皂苷Rg3浓度为41.6μmol/L时能抑制细胞迁移,促进细胞凋亡,并能够阻滞G₂/M期,上调凋亡相关蛋白表达,并且抑制STAT3信号通路相关蛋白表达,抑制STAT3蛋白的入核(P<0.05)。结论人参皂苷Rg3可能通过下调STAT3信号通路,从而抑制乳腺癌细胞MDA-MB-453的增殖及迁移水平,并促进其发生细胞凋亡。     

蛇葡萄素钠协同卡铂抑制人肺腺癌SPC-A-1细胞增殖

目的:探讨蛇葡萄素钠(AMP-Na)单用及其与卡铂(CBP)合用对SPC-A-1人肺腺癌细胞增殖的抑制作用及可能的机制。方法:采用MTT比色法测定AMP-Na单用及其与CBP合用对SPC-A-1细胞的体外细胞毒活性;用流式细胞仪分析AMP-Na单用及与卡铂合用后对SPC-A-1细胞凋亡的影响。结果:体外细胞毒实验和流式细胞仪结果表明,AMP-Na对人肺腺癌SPC-A-1细胞增殖的抑制作用具有明显的量效关系,与CBP联用后凋亡率较卡铂单用明显上升,有协同抑制作用。结论:AMP-Na单用能够显著抑制人肺腺癌SPC-A-1细胞的增殖,与卡铂合用可以增强卡铂的活性,协同抑制SPC-A-1细胞的增殖;AMP-Na可能是通过诱导凋亡而发挥其抗肿瘤作用。     

丹皮酚在体外对4种肿瘤细胞株的增殖抑制作用

目的探讨丹皮酚在体外对肿瘤细胞增殖的影响.方法采用MTT法检测丹皮酚对体外培养的肿瘤细胞的增殖抑制作用.结果丹皮酚在7.81～250 mg·L-1剂量下对体外培养的人红白血病细胞系K562、乳腺癌基因细胞系T6～17、肝癌细胞系Bel-7404及宫颈癌细胞系HeLa的增殖均有抑制作用.药物浓度越高,抑制作用越强,有明显的剂量效应关系.对不同的细胞系抑制作用不同,其IC50值分别为:30.83(K562)、13.44 (T6～17)、69.89 (Bel-7404)及93.91 mg·L-1(HeLa).以丹皮酚125及250 mg·L-1分别作用于以上细胞,于24、48、72及96 h观察细胞的生长情况,发现丹皮酚对细胞的增殖抑制作用有明显的时间效应关系,作用时间越长,抑制作用越强.结论丹皮酚在体外对多种肿瘤细胞具有增殖抑制作用.     

氯氧喹对不同类型乳腺癌细胞系的抑制作用及机制

目的探究氯氧喹对不同类型乳腺癌细胞增殖、凋亡及细胞周期阻滞的作用及其可能机制。方法采用不同浓度氯氧喹处理乳腺癌Bcap37、MDA-MB-231和MDA-MB-453细胞。MTT法检测细胞活力,Annexin V-/PI双染法采用流式细胞仪检测细胞凋亡和细胞周期,蛋白印迹法检测凋亡和细胞周期相关蛋白水平的变化。结果氯氧喹在50 mg·L~(-1)以上对3种细胞系的生长均有明显抑制作用(P<0.05),并呈剂量依赖性,且200 mg·L~(-1)氯氧喹与40 mg·L~(-1)紫杉醇作用相近。氯氧喹可明显诱导3种细胞凋亡(P<0.05),且对MDA-MB-231细胞凋亡的作用尤其明显(P<0.01),并与凋亡相关蛋白cleaved-PARP、cleaved-caspase的表达相一致。氯氧喹还可明显诱导3种细胞G_2/M期的数目增加(P<0.05),但在Bcap37和MDA-MB-453细胞更明显,并伴随p21蛋白下调和CDC2蛋白的上调。结论氯氧喹能有效抑制乳腺癌细胞的增殖活力,在MDA-MB-231细胞以促进凋亡为主,而在Bcap37和MDA-MB-453细胞以细胞周期阻滞为主。     

低分子量肝素联合吉西他滨对乳腺癌细胞侵袭和迁移能力的影响

目的探讨低分子量肝素(low molecular weight hepa-rin,LMWH)对吉西他滨(gemcitabine,GEM)抗肿瘤作用的影响及其作用机制。方法实验分为对照组、LMWH组、GEM组、LMWH与GEM联合应用组。MTT法测定药物对MDA-MB-231、MDA-MB-435细胞增殖的影响;细胞划痕实验、Tr-answell小室法检测细胞的迁移和侵袭能力;Western blot检测基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的蛋白表达;ELISA法检测乳腺癌细胞培养液中乙酰肝素酶(heparanase,HPA)的表达。结果 LMWH对MDA-MB-231、MDA-MB-435细胞的增殖没有明显影响,亦不能增强GEM的增殖抑制作用。30×104IU·L-1LMWH与2.5 mg·L-1GEM联合作用于MDA-MB-231、MDA-MB-435细胞24 h的迁移抑制率分别为66.3%、76.5%,较单用GEM的迁移抑制率(MDA-MB-231细胞47.4%,MDA-MB-435细胞52.9%)明显提高。同时LMWH与GEM合用对乳腺癌细胞侵袭的抑制也明显高于GEM单用时的作用。GEM可下调MMP-9和HPA的表达,与LMWH合用时下调更加明显。结论 LM-WH具有增强GEM抑制乳腺癌细胞迁移和侵袭的作用,其机制可能与下调MMP-9和HPA的表达有关。     

杠柳毒苷体外抑制肝癌细胞和乳腺癌细胞增殖的实验研究

目的探讨杠柳毒苷在体外对人乳腺癌MDA—MB．468细胞和人肝癌HepG2细胞增殖的影响。方法MTT法观察杠柳毒苷对人乳腺癌MDA-MB-468细胞和人肝癌HepG2细胞增殖的抑制作用,流式细胞术观察杠柳毒苷对两种肿瘤细胞的细胞增殖周期作用。结果与对照组比较,杠柳毒苷能明显抑制两种肿瘤细胞的增殖,其抑制率与药物浓度和作用时间呈正相关。流式细胞仪检测发现,杠柳毒苷对乳腺癌MDA-MB-468细胞和肝癌HepG2细胞持续作用24h后,可以使GdG1期细胞增多,G2／M期细胞减少。结论杠柳毒苷具有抑制乳腺癌MDA-MB-468细胞和肝癌HepG2细胞增殖的作用,并可将乳腺癌MDA-MB-468细胞和肝癌HepG2细胞的细胞生长周期阻滞在G0／G1期。     

金雀黄素对人乳腺癌细胞系体外增殖作用的影响

为了探讨金雀黄素对人乳腺癌细胞系体外增殖作用的影响,应用体外细胞培养和MTT比色法观察金雀黄素对人乳腺癌细胞系体外增殖作用的影响。结果表明,金雀黄素对MDA-MB-231细胞和MCF-7细胞的增殖均有抑制作用,但对MCF-7细胞抑制作用更为明显。结论:金雀黄素对乳腺癌细胞系体外增殖具有明显的抑制作用,为进一步阐明其抑癌作用机制奠定了实验基础。     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Polymorphisms in genes involved in neurotransmission in relation to smoking

Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D₁, D₂, and D₄ receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D₃ and D₅ receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking.     

Tramadol concentrations in blood and in cerebrospinal fluid in a neonate

Based on blood and cerebrospinal fluid samples collected in a full-term neonate, the penetration of tramadol in the central nervous system is described. Following intravenous administration of tramadol, a lag time of about 4 h was observed until full blood–brain equilibration was achieved. This pharmacokinetic observation is in line with a recent pharmacodynamic evaluation of the central opioid effects of tramadol in adults.     

Disease control in asthmatic children seen in private practice in Switzerland

ABSTRACT

Background: Asthma is the most common chronic childhood disease in Switzerland with a prevalence of 10%. Asthma has a high economic burden accounting for high medical costs. Assessment of disease control is likely to be of help in the implementation of strategies to improve asthma. Therefore, we aimed to evaluate asthma control and therapy regimens among children in private practice.

Methods: We assessed asthma control as well as therapy regimens in 575 asthmatic children in an experience programme in Switzerland by using an abbreviated questionnaire based on the asthma control questionnaire and the child health questionnaire on Visit 1 and Visit 2.

Results: Good asthma control at Visit 1 was only present in 25.7% of asthmatic children. Occasional asthma symptoms, limitation of physical activity, nocturnal awakening and anxiety of the parent was present in 80.5%, 41.2%, 46.8% and 57% of the children, respectively. After adjustment of therapy regimens at Visit 1, mainly by adding a leukotriene receptor antagonist, asthma control was reported to be much better in 53.4% of the children at Visit 2.

Conclusions: As asthma control is inadequately achieved within a major portion of asthmatic children, it is imperative to find measures to improve asthma control and hence, to reduce the burden of disease.