PI3K抑制剂对肺癌AKT2、MMP9表达的抑制作用的研究及意义

目的探讨PI3K抑制剂LY294002对肺腺癌AKT2及基质金属蛋白酶9(MMP9)表达的作用及意义。方法不同浓度(5μmol/L、10μmol/L、20μmol/L、40μmol/L)LY294002处理肺腺癌A549细胞24h后,分别用免疫细胞化学法和Western blot法检测AKT2、MMP9蛋白表达。人肺癌A549细胞接种于裸鼠皮下,建立肺癌裸鼠移植瘤模型并予LY294002治疗,计算LY294002的抑瘤率,Western blot法测定裸鼠移植瘤组织AKT2、MMP9蛋白的表达。结果肺腺癌细胞株A549细胞中存在AKT2、MMP9蛋白的高表达,应用P13K抑制剂LY294002可抑制该细胞株细胞中AKT2、MMP9蛋白的表达,而且其表达呈浓度依赖型降低。LY294002可有效抑制裸鼠移植瘤的生长,其抑瘤率为40.30%。LY294002治疗组裸鼠移植瘤组织MMP9及VKT2蛋白水平均显著低于对照组(P均<0.01)。结论 LY294002可抑制AKT2活性及下调MMP9蛋白的表达,该作用可能是PI3K抑制剂LY294002发挥抗肿瘤生长及及转移的机制之一。PI3K可能成为肺癌治疗的靶点。     

尼美舒利对人舌鳞癌裸鼠移植瘤环氧化酶-2的影响

目的研究环氧化酶-2(COX-2)抑制剂尼美舒利对人舌鳞癌裸鼠移植瘤的抑制作用,分析尼美舒利对COX-2基因及蛋白表达的影响,进一步探讨其抑癌机制。方法取人舌鳞癌Tca-8113细胞接种于16只裸鼠皮下,建立人舌鳞癌裸鼠移植瘤模型,随机分为对照组和尼美舒利治疗组,计算尼美舒利的抑瘤率。反转录-聚合酶链反应(RT-PCR)检测裸鼠移植瘤组织COX-2mRNA表达。结果尼美舒利治疗组抑瘤率为47.2%。尼美舒利治疗组裸鼠移植瘤组织COX-2mRNA水平均低于对照组,差异有统计学意义(P<0.05)。结论 COX-2抑制剂尼美舒利可下调COX-2基因表达,该作用可能是COX-2抑制剂抑制肿瘤血管生成从而抑制肿瘤生长的机制之一。     

IGF1R/PI3K/Akt通路及其调控对非小细胞肺癌血管生成拟态形成影响的研究

目的 为非小细胞肺癌(NSCLC)的治疗探索一个潜在的分子靶标。方法 通过qPCR、Western blot、免疫组化检测NSCLC组织和癌旁组织KLF16、IGF1R表达情况；通过抑制IGF1R/PI3K/Akt通路活化，检测相关靶标MMP2和MMP9的表达；通过IGF1R/PI3K/Akt通路抑制剂LY294002干预血管生成拟态(VM)形成。结果 与癌旁组织相比，免疫组化、Western blot和qPCR提示，KLF16在NSCLC组织中表达降低，且随着NSCLC级别提高，表达量递减。NSCLC组织中存在明显IGF1R阳性，且IGF1R阳性表达量随着NSCLC级别增高而递增。同时，Western blot和Matrigel三维细胞培养结果表明，随着IGF1R/PI3K/Akt通路抑制剂浓度增加，IGF1R/PI3K/Akt通路相关蛋白质、MMP2、MMP9表达水平也随之降低，肿瘤VM形成随之减少。结论 NSCLC VM的形成与IGF1R/PI3K/Akt信号通路有关，KLF16在NSCLC中的表达趋势与IGF1R表达相反，两者之间是否存在调控关系需进一步验证。     

PI3K信号通路在LPS诱导RAW264.7细胞表达sTREM-1中的作用

目的：探讨磷脂酰肌醇3-激酶(PI3K)信号通路在脂多糖(LPS)诱导RAW264.7细胞表达可溶性髓样细胞触发受体-1(sTREM-1)中的作用。方法培养小鼠巨噬细胞株RAW264.7,采用相同浓度的LPS在不同时间诱导RAW264.7细胞,应用Western blot法分别检测PI3K蛋白表达水平,RT-PCR法检测PI3K mRNA表达水平,酶联免疫吸附(ELISA)法检测细胞培养血清中sTREM-1表达水平。用不同浓度PI3K特异性抑制剂LY294002处理细胞,观察上述指标变化。结果 LPS可时间依赖性地诱导RAW264.7细胞PI3K蛋白、PI3K mRNA的表达;LY294002可浓度依赖性地抑制PI3K蛋白、PI3K mRNA的表达;LY294002阻断PI3K信号转导通路后,LPS对sTREM-1表达的诱导作用受到显著抑制,并且具有剂量依赖性。结论 LPS通过PI3K信号通路诱导RAW264.7细胞表达sTREM-1。     

舒脉汤对血管内皮细胞VEGF、bFGF表达的影响及其信号通路

目的研究舒脉汤对血管内皮细胞生成因子（VEGF）、碱性成纤维细胞生成因子（bFGF）的影响及其可能参与的信号通路。方法原代培养人脐静脉内皮细胞,以舒脉汤刺激内皮细胞,部分给予PI3K抑制剂,LY294002预处理,收集细胞测定VEGF、bFGF基因表达,收集上清液测定VEGF、bFGF蛋白表达。结果舒脉汤可以明显增加人脐静脉内皮细胞VEGF、bFGF基因及蛋白表达,且这一过程可被LY294002所阻断。结论舒脉汤通过上调内皮细胞VEGF、bFGF表达来发挥促血管新生作用,PI3K信号通路参与此过程。     

异氟醚对小儿神经母细胞瘤术后肿瘤复发及转移的影响

目的 探究异氟醚对小儿神经母细胞瘤术后肿瘤复发和转移的影响。方法 采用ELISA法检测经异氟醚麻醉前后神经母细胞瘤患者血清中上皮间质转化(epithelial mesenchymal transition, EMT)相关蛋白E-钙粘蛋白(E-cadherin)、波形蛋白(vimentin)和血管内皮因子(vascular endothelial growth factor, VEGF)的表达水平；采用CCK-8、Transwell和Western blotting法检测经异氟醚或同时经PI3K抑制剂LY294002处理的SH-SY5Y细胞增殖、迁移、侵袭及EMT和PI3K/Akt通路相关蛋白、VEGF和HIF-1α的表达水平。结果 在经异氟醚麻醉的神经母细胞瘤患者血清中E-cadherin表达下调，Vimentin和VEGF表达上调。异氟醚能够促进SH-SY5Y细胞增殖、迁移、侵袭和EMT,并上调p-PI3K、p-Akt和HIF-1α的表达水平；异氟醚通过激活PI3K/Akt通路上调HIF-1α的表达促进SH-SY5Y细胞生物学行为，且能够被PI3K抑制剂LY294002阻断。结论 异...     

LY294002对类脂A诱导的人冠状动脉内皮细胞缝隙连接蛋白43表达的影响

目的 探讨LY294002对类脂A(KLA)诱导的人冠状动脉内皮细胞(HCAEC)缝隙连接蛋白43(Cx43)表达的影响。方法 用CCK8法检测0、0.01、0.05、0.10、0.20 mg/L KLA和0、2.5、5.0、10.0、20.0μmol/L磷酸肌醇3激酶(PI3K)特异性抑制剂——LY294002对HCAEC活力的影响。将HCAEC采用简单随机分组法分为空白对照组(Con组)、Toll样受体4(TLR4)激动组(KLA组)、PI3K抑制组(LY组)和KLA+LY组，采用Western Blot法检测各组HCAEC TLR4、PI3K、Cx43蛋白表达水平。结果 0.01、0.05 mg/L KLA HCAEC存活率与0 mg/L比较，2.5μmol/L LY294002 HCAEC存活率与0μmol/L比较，差异均无统计学意义(P>0.05);0.10、0.20 mg/L KLA HCAEC存活率均明显低于0 mg/L,5.0、10.0、20.0μmol/L LY294002 HCAEC存活率均明显低于0μmol/L,差异均有统计学意义(P<0.05)。KL...     

力达霉素诱导人胃癌BGC823细胞凋亡和抑制裸鼠移植瘤生长

观察力达霉素(LDM)对人胃癌BGC823细胞的诱导凋亡作用及体内抗肿瘤活性。采用MTT法观察LDM对人胃癌BGC823细胞增殖的抑制作用。利用Annexin V-FITC/PI双染结合流式细胞仪和脱氧核糖核酸末端转移酶介导的缺口末端标记技术检测细胞凋亡的改变。采用Western blotting法检测细胞中VEGF蛋白的表达情况。建立裸鼠胃癌皮下移植瘤模型，观察LDM的体内抗肿瘤活性。LDM能够明显抑制BGC823细胞增殖，诱导细胞凋亡，降低细胞VEGF蛋白的表达，抑制胃癌裸鼠移植瘤的生长。LDM剂量0.02和0.04 mg·kg^-1的抑瘤率分别为57%和72%(P<0.01)。LDM可诱导胃癌细胞凋亡并抑制裸鼠移植肿瘤的生长。     

青蒿琥酯对人乳腺癌MCF-7血管生成和肿瘤浸润作用研究

目的:观察青蒿琥酯对人乳腺癌MCF-7血管生成及浸润转移的抑制作用.方法:建立人乳腺癌MCF-7裸鼠移植瘤模型,给予不同剂量青蒿琥酯治疗并观察其对移植瘤的抑制作用;用免疫组化检测移植瘤组织细胞中VEGF、HIF-1α、uPA、E-cadherin蛋白表达和Western blot检测移植瘤组织细胞中HIF-lα、VEGF蛋白表达.结果:青蒿琥酯低、高剂量组抑瘤率分别为(24.39±10.20)％、(40.24±7.02)％;免疫组化结果显示,与生理盐水组比较,青蒿琥酯低、高剂量组VEGF、HIF-1α蛋白表达显著降低,差异具有统计学意义(P＜0.05);uPA和E-cadherin蛋白在生理盐水组和青蒿琥酯低、高剂量组中的表达水平差异均无统计学意义(P＞0.05),HIF-1 α/VEGF之间表达呈正相关(r =0.983,P=0.000);West-em blot结果显示,青蒿琥酯低、高剂量组中VEGF、HIF-1α蛋白下调,表达量低于生理盐水组,青蒿琥酯高剂量组的蛋白表达量最低.HIF-lα与VEGF蛋白表达之间存在一致性.结论:青蒿琥酯可显著抑制乳腺癌裸鼠移植瘤的生长,其机制可能与抗肿瘤血管及抑制肿瘤侵袭相关.     

组蛋白去乙酰化酶抑制剂抗血管新生机制的体内研究

目的 探讨组蛋白去乙酰化酶(HDAC)抑制剂对白血病细胞移植瘤血管新生的影响及其机制.方法 12只裸鼠均分为对照组和丙戊酸钠(VPA)用药组.建立Kasumi-1细胞裸鼠移植瘤模型,应用HDAC抑制剂VPA体内用药.RT-PCR和免疫组化检测血管内皮生长因子(VEGF)及其受体(VEGFR) mRNA或蛋白的表达;染色质免疫共沉淀法检测VEGF基因启动子区域染色质内组蛋白H3乙酰化程度的变化.结果 与对照组比较,VPA用药组VEGF及VEGFR2 mRNA和蛋白表达水平明显减少,启动子区域染色质内组蛋白H3的乙酰化程度明显升高.结论 VPA作为HDAC抑制剂,通过提高组蛋白的乙酰化程度,进而抑制血管新生相关因子及受体的表达,阻碍白血病的血管新生.     

Role of glutathione in reduction of arsenate and of gamma-glutamyltranspeptidase in disposition of arsenite in rats

Arsenate (AsV), the environmentally prevalent form of arsenic, is converted sequentially in the body to arsenite (AsIII), monomethylarsonic acid (MMAsV), monomethylarsonous acid (MMAsIII), and dimethylarsinic acid (DMAsV) and some trimethylated metabolites. Although the biliary excretion of arsenic in rats is known to be glutathione (GSH)-dependent, involving transport of arsenic-GSH conjugates, the role of GSH in the reduction of AsV to the more toxic AsIII in vivo has not been defined. Therefore, we studied how the fate of AsV is influenced by buthionine sulfoximine (BSO), which depletes GSH in tissues. Control and BSO-treated rats were given AsV (50 micromol/kg, i.v.) and arsenic metabolites in bile, urine, blood and tissues were analysed by HPLC-HG-AFS. BSO increased retention of AsV in blood and tissues and decreased appearance of AsIII in blood, bile (by 96%) and urine (by 63%). The biliary excretion of MMAsIII was also nearly abolished, the appearance of MMAsIII and MMAsV in the blood was delayed and the renal concentrations of these monomethylated arsenicals were decreased by BSO. Interestingly, appearance of DMAsV in blood and urine remained unchanged and the concentrations of this metabolite in the kidneys and muscle were even increased in response to BSO. To test the role of gamma-glutamyltranspeptidase (GGT) in arsenic disposition, the effect of the of the GGT inhibitor acivicin was investigated in rats injected with AsIII (50 micromol/kg, i.v.). Acivicin lowered the hepatic and renal GGT activities and increased the biliary as well as urinary excretion of GSH, but failed to alter the disposition (i.e. blood and tissue concentrations, biliary and urinary excretion) of AsIII and its metabolites. In conclusion, shortage of GSH decreases not only the hepatobiliary transport of arsenic, but also reduction of AsV and the formation of monomethylated arsenic, while not hindering the production of dimethylated arsenic. While GSH plays an important role in the disposition and toxicity of arsenic, GGT, which hydrolyses GSH and GSH conjugates, apparently does not influence the fate of the GSH-reactive trivalent arsenicals in rats.     

微透析取样技术在中药体内分析中的应用研究进展

本文综述了微透析取样技术在中药体内分析中的应用,介绍微透析取样技术的原理、组成、探针类型、特点,重点阐述了微透析取样技术在测定脑、血液、皮肤等组织器官中中药有效成分浓度的应用实例。表明微透析取样技术在中药药效研究中具有广阔的前景。     

2009年某院住院患者抗真菌药用药调查

目的:分析讨论某院抗真菌药使用的合理性,为临床安全有效地使用抗真菌药提供参考。方法:回顾性统计分析某院2009年住院患者抗真菌药用药信息。结果:2009年某院住院患者抗真菌药DDDs排名前3名分别为:氟康唑、制霉菌素和伊曲康唑;使用金额排名前3名分别为:氟康唑、米卡芬净及卡泊芬净;更换一种抗真菌药进行治疗的患者数为176人,在全部患者中占13.4%。结论:应进一步强化用药指征的意识,提高标本送检率,同时改善某些抗真菌用药不合理更换的现象,以避免耐药性发生,从而更好更长远地体现抗真菌药的治疗价值。     

Role of desacetylation in the detoxification of cephalothin in renal cells in culture

The toxicity of three cephalosporin antibiotics to rabbit kidney cells in culture was compared to their known nephrotoxic potential in vivo (cephaloridine greater than cefazolin greater than cephalothin). While cephalothin is considered to be a relatively nonnephrotoxic cephalosporin when administered to many species including humans and rabbits, in several in vitro systems involving rabbit renal tissue, cephalothin was comparatively more toxic than anticipated based on in vivo data. Cephalothin is extensively desacetylated in rabbits to a less microbiologically active metabolite, desacetylcephalothin. When a microsomal S9 fraction from rabbit kidney was added to the in vitro assay in cultured rabbit renal cells, cephalothin was desacetylated and its toxicity to kidney cells was reduced. The addition of S9 in vitro provided a toxicity ranking of the cephalosporins that correlated with their known in vivo nephrotoxic potentials (cephaloridine greater than cefazolin greater than cephalothin). The in vitro detoxification of cephalothin by S9 was blocked by the coadministration of the esterase inhibitor, aminocarb. Desacetylcephalothin was relatively nontoxic to rabbit renal tissue in vitro. These results suggest that the desacetylation of cephalothin in vivo represents a previously unrecognized mechanism of detoxification of this cephalosporin antibiotic. Furthermore, this mechanism of detoxification may be applicable to other acetylated cephalosporins.     

应用PASS系统对我院2008年住院医嘱的分析

目的监测分析2008年我院住院患者用药情况。方法将PASS系统嵌入医生工作站、临床药学工作站等子系统,构建合理用药计算机网络系统,对住院医嘱进行及时监测,将监测结果向医生反馈,并对其进行统计、分析。结果2008年共监测医嘱3 620 241条,不合理医嘱908条,占0.02%。不合理医嘱中,配伍禁忌(381条)占41.96%,用法用量(381条)占41.96%,药物相互作用(108条)占11.89%,儿童用药(38条)占4.19%。经与医生沟通后,更改不合理医嘱856条,占94.27%。结论PASS系统可有效监测医嘱中的不合理用药,通过与医生交流,大大减少药物不良事件的发生,值得临床推广应用,也为临床药师开展工作带来了极大的便利。但PASS系统尚存在局限性,有待进一步完善。     

应用PASS系统分析我院2010年住院医嘱

目的:了解我院2010年住院患者的合理用药情况,探讨如何利用合理用药监测系统( PASS)提高合理用药水平.方法:利用PASS对我院2010年15 966例住院患者的1 184 997条用药医嘱进行监测,以黑色警示医嘱为依据,收集不合理用药信息,并对监测结果进行统计、分析.结果:不合理用药医嘱50 261条,发生率为4.24％.绝对禁止黑色医嘱5441条,主要为药物相互作用(66.54％)、注射液体外配伍(17.86％)、用法用量(15.46％)、儿童警告(1.14％).结论:应用PASS系统能有效监测医嘱中的不合理用药情况,有利于提高临床合理用药水平,但PASS系统尚存在局限性,有待进一步完善.     

Kinetics of in vitro metabolism of methoxyphenamine in rats

1. Methoxyphenamine (MP) was metabolized in vitro by rat liver preparations to O-desmethylmethoxyphenamine (O-desmethyl-MP), N-desmethylmethoxyphenamine (N-desmethyl-MP) and 5-hydroxymethoxyphenamine (5-hydroxy-MP). These metabolic pathways were inhibited by SKF 525-A and carbon monoxide, which indicates that these reactions were mediated at least partly by an NADPH-dependent cytochrome P-450 system. 2. Strain differences in the metabolism of this drug in vitro were observed in female Lewis and Dark Agouti (DA) rats, which are proposed models for human debrisoquine phenotypes. Methoxyphenamine O-demethylase and 5-hydroxylase activity in DA rats were lower than those in Lewis rats. 3. The metabolic transformation of methoxyphenamine in vitro to O-desmethyl-MP was inhibited competitively by debrisoquine and sparteine. This indicates that the cytochrome P-450 isoenzyme mediating the metabolism of MP to O-desmethyl-MP is similar to that mediating metabolism of debrisoquine and sparteine. However, no inhibition was observed with methenytoin.     

Changes in levels of dependency and predictors of mortality in elderly people in institutional care in Dunedin

The 1983 study of dependency of subjects in institutional care in Dunedin was repeated two years later. A significant increase in levels of dependency in residential homes, particularly in the Religious and Welfare sector was found. In 1983 there were 29 high dependency residents and 73 medium dependency residents in residential homes. In 1985 these numbers had increased to 55 and 86 respectively. There was no change in the number of low dependency residents. In 1983, 6 high dependency residents had been admitted to residential home care in the year prior to the study. In 1985 the number of high dependency residents recently admitted had increased to 23. There had also been a significant increase in the dependency of patients in Religious and Welfare continuing care hospitals. Of the 933 subjects in institutional care in 1983 who were able to be followed, 354 (37.9%) died in the following 2 years. Mortality rate was higher for those in hospital care (48.1%) than for those in residential home care (29.6%). Mortality rates were higher in more dependent subjects and this was evident for each measure of dependency.     

基层医院护士在临床合理用药中的作用

目的充分利用护士在医师和患者间的特殊地位和作用,促进基层临床合理用药。方法从护士的工作性质出发,论述护士参与促进合理用药的方便和优势。结果通过实践,护士在促进合理用药中的作用得到有效发挥,基层合理用药环境得到极大改善。结论充分利用护士与医师和患者间的特殊桥梁作用,在基层医院促进合理用药,规范医师用药行为,防止药物滥用,引导患者安全用药,降低药源性疾病。