c-Met信号通道参与HGF诱导不同基因型非小细胞肺癌细胞株对吉非替尼耐药

背景与目的肝细胞生长因子(hepatocyte growth factor,HGF)诱导非小细胞肺癌(non-small cell lung cancer,NSCLC)对吉非替尼耐药,可能与其受体c-Met激活有关。本研究旨在探讨c-Met及其下游信号通道是否参与HGF诱导不同基因型NSCLC细胞株对吉非替尼耐药。方法选择人NSCLC细胞株表皮生长因子受体(epidermal growth factor receptor,EGFR)突变型PC-9、PC9/R和EGFR野生型H292、A549,用HGF诱导细胞,通过MTT法检测细胞增殖,Annexin V-FITC法检测细胞凋亡,应用免疫印迹技术检测细胞中c-Met及下游通道的变化。结果吉非替尼对PC9、H292、A549的生长抑制作用呈浓度依赖性,HGF诱导后吉非替尼抑制细胞的生长曲线明显往右移。在PC9、H292、A549细胞中,吉非替尼和HGF处理组的细胞凋亡率比吉非替尼处理组均减少(P<0.05),在PC9/R细胞中无明显减少(P>0.05)。HGF能激活PC9、H292、PC9/R、A549细胞中c-Met及其下游通道蛋白。在PC9、H292、A549细胞中,吉非替尼和HGF处理组的p-Met、p-Akt、p-Stat3、p-Erk1/2蛋白表达比吉非替尼处理组均增高,在PC9/R细胞中无明显增高。结论在体外HGF诱导不同基因型NSCLC细胞株对吉非替尼耐药,c-Met及其下游信号通道参与HGF诱导不同基因型NSCLC细胞株对吉非替尼耐药。     

肝细胞生长因子诱导不同基因型非小细胞肺癌细胞株对埃罗替尼耐药及机制的研究

背景与目的肝细胞生长因子(hepatocyte growth factor ,HGF)受体(c-Met)可能与非小细胞肺癌(non-small cell lung cancer,NSCLC)对埃罗替尼耐药有关。本研究旨在体外研究HGF诱导不同EGFR基因型NSCLC细胞对埃罗替尼耐药及可能的耐药机制。方法 选择不同EGFR基因型NSCLC细胞(PC9、H292、A549),用HGF诱导或(和)埃罗替尼处理细胞,通过MTT法检测细胞增殖,PI法检测细胞周期,Annexin V-FITC法检测细胞凋亡,Western blotting检测细胞中c-Met、EGFR、ErbB3及其磷酸化蛋白的表达。结果 埃罗替尼对三种细胞的生长抑制作用均呈浓度依赖性,埃罗替尼对HGF诱导的药物-存活率曲线明显往右移。实验分为四组：C组(control)、H组(HGF)、E组(erlotinib)、HE组(HGF erlotinib)。在每种细胞分组中,HE组凋亡率均比E组减少(P<0.05)。在H292、A549,HE组与E组比较,G0/G1期比例减少(P<0.05)、S期比例增多(P<0.05)。HGF能迅速活化三种细胞内p-Met蛋白磷酸化,HE组与E组比较,p-Met蛋白含量明显升高。结论 在体外,在体外HGF诱导不同EGFR基因型NSCLC细胞株对埃罗替尼耐药,c-Met磷酸化活化可能是HGF诱导NSCLC细胞株对埃罗替尼耐药的重要机制。     

SU11274逆转肝细胞生长因子诱导不同EGFR基因型非小细胞肺癌细胞株对吉非替尼耐药

背景与目的:肝细胞生长因子(hepatocyte growth factor,HGF)诱导敏感非小细胞肺癌(nonsmall cell lung cancer,NSCLC)细胞对表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitor,EGFR-TKI)耐药,其机制与c-Met激活有关。本研究探讨c-Met抑制剂SU11274逆转HGF诱导的不同EGFR基因型NSCLC细胞株对吉非替尼耐药及逆转耐药机制。方法:选择人NSCLC细胞株PC9(EGFR突变型)、H292(EGFR野生型)和A549(EGFR野生型),应用吉非替尼和SU11274单独或联合作用于HGF诱导的细胞株。实验分为6组:C组(不加药对照组)、H组(HGF处理组)、G组(吉非替尼处理组)、S组(SU11274处理组)、HG组(HGF+吉非替尼处理组)和HGS组(HGF+吉非替尼+SU11274处理组)。MTT法检测对细胞增殖的影响,流式细胞术检测细胞凋亡的影响;应用蛋白质印迹法(Western blot)检测细胞中c-Met及其下游通道Stat3、Akt和Erk1/2蛋白表达水平。结果:吉非替尼对3种细胞的生长抑制作用均呈浓度依赖性,HGF处理能够缓解吉非替尼的增殖抑制作用(P<0.05);不同浓度吉非替尼联合SU11274作用于HGF诱导细胞时,3种细胞株存活率比吉非替尼单独作用于HGF诱导细胞时明显降低(P<0.05);HGS组的细胞凋亡比HG组明显增加(P<0.05);HGS组的c-Met、Stat3、Akt和Erk1/2活化蛋白量比HG组明显减少。结论:c-Met抑制剂SU11274可逆转HGF诱导的不同EGFR基因型NSCLC细胞株对吉非替尼耐药,其机制可能与抑制HGF活化的c-Met及其下游通道蛋白表达有关。     

SU11274逆转肝细胞生长因子诱导不同EGFR基因型非小细胞肺癌细胞株对吉非替尼耐药

背景与目的：肝细胞生长因子(hepatocyte growth factor,HGF)诱导敏感非小细胞肺癌(nonsmall cell lung cancer,NSCLC)细胞对表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitor,EGFR-TKI)耐药,其机制与c-Met激活有关。本研究探讨c-Met抑制剂SU11274逆转HGF诱导的不同EGFR基因型NSCLC细胞株对吉非替尼耐药及逆转耐药机制。方法：选择人NSCLC细胞株PC9(EGFR突变型)、H292(EGFR野生型)和A549(EGFR野生型),应用吉非替尼和SU11274单独或联合作用于HGF诱导的细胞株。实验分为6组：C组(不加药对照组)、H组(HGF处理组)、G组(吉非替尼处理组)、S(SU11274处理组)、HG组(HGF+吉非替尼处理组)和HGS组(HGF+吉非替尼+SU11274处理组)。MTT法检测对细胞增殖的影响,流式细胞术检测细胞凋亡的影响;应用蛋白质印迹法(Western blot)检测细胞中c-Met及其下游通道Stat3、Akt和Erk1/2蛋白表达水平。结果：吉非替尼对3种细胞的生长抑制作用均呈浓度依赖性,HGF处理能够缓解吉非替尼的增殖抑制作用(P<0.05);不同浓度吉非替尼联合SU11274作用于HGF诱导细胞时,3种细胞株存活率比吉非替尼单独作用于HGF诱导细胞时明显降低(P<0.05);HGS组的细胞凋亡比HG组明显增加(P<0.05);HGS组的c-Met、Stat3、Akt和Erk1/2活化蛋白量比HG组明显减少。结论：c-Met抑制剂SU11274可逆转HGF诱导的不同EGFR基因型NSCLC细胞株对吉非替尼耐药,其机制可能与抑制HGF活化的c-Met及其下游通道蛋白表达有关。     

HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄洛替尼的耐药

目的：探究肝细胞生长因子（hepatocyte growth factor,HGF） 体外诱导不同EGFR基因型非小细胞肺癌（non-small cell lung cancer,NSCLC）细胞对厄洛替尼的耐药及其可能的机制。方法：用HGF、厄洛替尼单独或联合处理人NSCLC细胞株PC9（EGFR突变型,敏感株）、H292（EGFR野生型,敏感株）、A549（EGFR野生型,原发性耐药株）,实验分为四组：C组（不加药对照组）、H组（HGF处理）、E组（厄洛替尼处理组）、HE组（HGF+厄洛替尼联合处理组）。MTT法检测其对细胞增殖的影响,流式细胞术检测其对细胞周期和凋亡的影响,Western blotting检测其对细胞中c-Met、EGFR、ErbB3及其磷酸化蛋白表达的影响。结果：厄洛替尼对3种细胞增殖抑制的作用均呈浓度依赖性,HGF处理能够缓解厄洛替尼对瘤细胞增殖的抑制作用。3种细胞的HE组凋亡率均显著低于E组（均P<0.05）。厄洛替尼阻滞3种细胞周期于G1期,对于H292、A549细胞,HE组G0/G1期比例显著低于E组（P<0.05）。HE组p-Met蛋白含量较E组显著升高（P<0.05）,而p-EGFR和p-ErbB3表达无显著差异（P>0.05）。结论：在体外,HGF能够诱导不同EGFR基因型NSCLC细胞株对厄洛替尼耐药,其机制可能与其诱导c-Met磷酸化活化有关。     

c-Met抑制剂SU11274在HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄罗替尼耐药的逆转作用

目的： 肝细胞生长因子(hepatocyte growth factor, HGF)诱导敏感非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞对表皮生长因子受体酪氨酸激酶抑制剂（epidermal growth facter receptor tyrosine kinase inhibitor, EGFR-TKI）厄洛替尼耐药,本研究旨在探讨c-Met抑制剂SU11274逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞株对厄洛替尼耐药及逆转耐药机制。方法： 选择人NSCLC细胞株PC9（EGFR突变型,敏感株）、H292（EGFR野生型,敏感株）和A549（EGFR野生型,原发性耐药株）,应用厄洛替尼和SU11274（1 μmol /L）单独或联合作用于HGF（40 ng/ml）诱导的细胞株,实验分为6组：C组（不加药对照组）、H组（HGF处理）、E组（厄洛替尼处理组）、S组（SU11274处理组）、HE组（HGF+厄洛替尼处理组）、HES组（HGF+厄洛替尼+ SU11274处理组）。MTT法、流式细胞术分别检测不同药物处理对各细胞增殖、凋亡的影响;应用Western blotting检测不同药物处理对各细胞中c-Met及其下游通道Stat3、Akt、Erk1/2蛋白表达水平。结果：厄洛替尼对3种细胞的增殖抑制作用均呈浓度依赖性,HGF处理能够缓解厄洛替尼的增殖抑制作用（P<0.05）;不同浓度厄洛替尼联合SU11274作用于HGF诱导细胞时3种细胞株存活率比厄洛替尼单独作用于HGF诱导细胞时明显降低（P<0.05）;HGS组的细胞凋亡比HG组明显增加（P<0.05）;HGS组的c-Met、Stat3、Akt、Erk1/2活化蛋白量比HG组明显减少。结论： c-Met抑制剂SU11274和厄洛替尼联合应用可逆转HGF诱导不同EGFR基因型非小细胞肺癌细胞对厄洛替尼耐药,其机制可能与抑制HGF活化的c-Met及其下游通道蛋白表达有关。     

AHR活化介导的ROS诱导NSCLC细胞吉非替尼耐药的作用机制

目的:探讨芳香烃受体（AHR）介导的活性氧簇（ROS）诱导非小细胞肺癌（NSCLC）吉非替尼耐药的作用机制。方法:以正常支气管肺泡上皮细胞BEAS-2B为对照,Western blot检测非小细胞肺癌A549、PC-9、H1299和H1975细胞AHR的蛋白表达。AHR激动剂FICZ、吉非替尼、N-乙酰半胱氨酸（NAC）分别处理A549和PC-9细胞,MTT、激光共聚焦显微镜、流式细胞术及Western blot分别检测吉非替尼敏感性、细胞内ROS及表皮生长因子受体（EGFR）信号。结果:MTT检测显示FICZ通过上调半数最大抑制浓度（IC50）诱导PC-9及A549细胞对吉非替尼耐药,Western blot显示FICZ可导致PC-9及A549细胞EGFR、AKT磷酸化,而NAC可遏制A549细胞中的EGFR磷酸化并逆转吉非替尼耐药。共聚焦显微镜和流式细胞仪显示,FICZ诱导的AHR活化导致PC-9及A549细胞内ROS产生增加,并且可被NAC抑制。结论:活化的AHR可通过促进NSCLC细胞中的ROS产生和EGFR信号转导介导吉非替尼耐药,此过程可被NAC抑制。     

上皮-间质转化和胰岛素样生长因子Ⅰ型受体在非小细胞肺癌EGFR-TKIs获得性耐药中的作用

目的:探讨上皮-间质转化(epithelial-mesenchymal transition,EMT)和胰岛素样生长因子Ⅰ型受体(insulin-like growth factor Ⅰ receptor,IGF-1R)在非小细胞肺癌(non-small cell lung cancer,NSCLC)对表皮生长因子受体-酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitors,EGFR-TKIs)获得性耐药中的作用.方法:选用EGFR基因突变型和野生型的人肺腺癌细胞PC-9和H460,分别用吉非替尼和厄洛替尼将其诱导成耐药的PC-9/ZD和H460/ER细胞.MTT法检测各组细胞的增殖情况,划痕实验及Transwell侵袭实验检测细胞迁移和侵袭能力,蛋白质印迹法和RT-PCR法分别检测IGF-1R及EGFR信号通路分子及EMT相关分子的蛋白和mRNA表达.结果:PC-9/ZD和H460/ER为EGFR-TKI获得性耐药细胞,分别对吉非替尼和厄洛替尼的敏感性显著下降,差异有统计学意义(P＜0.05).与PC-9和H460细胞相比,耐药的PC-9/ZD和H460/ER细胞形态呈现间质细胞表型,细胞侵袭和迁移能力增加(P＜0.05).PC-9/ZD和H460/ER细胞中间质标志分子vimentin的mRNA和蛋白表达量均显著增加(P＜0.05),PC-9/ZD细胞的E-cadherin表达量显著减少(P＜0.05).与PC-9和H460细胞相比,PC-9/ZD与H460/ER细胞的IGF-1R及其磷酸化蛋白表达量显著增加(P＜0.05),AKT和ERK磷酸化水平明显上调(P＜0.05).PC-9/ZD细胞的EGFR磷酸化水平与PC-9细胞相比未见明显差异(P＞0.05),而H460/ER细胞的EGFR磷酸化水平较H460细胞显著降低(P＜0.05).结论:EGFR突变型PC-9细胞和野生型H460细胞的获得性耐药细胞中均发生了EMT,并且IGF-1R信号蛋白表达增多,提示EMT及IGF-1R信号转导通路均可能在NSCLC的EGFR-TKIs获得性耐药中起重要作用.     

洛伐他汀克服非小细胞肺癌PC9细胞株吉非替尼耐药的体外研究

目的:研究联合洛伐他汀(lovastatin)和吉非替尼(gefi tinib)对体外诱导吉非替尼获得性耐药的非小细胞肺癌细胞株PC9细胞凋亡以及相关蛋白表达的影响,并探讨其可能的机制。方法:应用洛伐他汀联合吉非替尼处理耐吉非替尼的非小细胞肺癌PC9细胞株后,采用WST-1法检测不同药物处理对PC9细胞增殖的影响,Hoechst33342荧光染色法观察细胞凋亡形态,FCM法观察细胞凋亡状况,蛋白质印迹法检测凋亡相关蛋白的表达水平。结果:洛伐他汀联合吉非替尼可在体外诱导耐吉非替尼的PC9细胞凋亡,抑制其细胞增殖;洛伐他汀联合吉非替尼可诱导耐吉非替尼的PC9细胞中磷酸化表皮生长因子受体(phosphorylated epidermal growth factor receptor,p-EGFR)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)和磷酸化细胞外调节蛋白激酶1/2(phosphorylated extracellular signal-regulated kinase1/2,p-ERK1/2)蛋白表达水平明显下调。结论:在体外诱导吉非替尼获得性耐药的非小细胞肺癌细胞株PC9中,洛伐他汀可以克服吉非替尼耐药,两者具有良好的协同作用,提示两药联合对于出现吉非替尼耐药的非小细胞肺癌的临床治疗可能具有很大的应用潜力。     

烟草烟雾通过ROS/Sirt3/SOD2通路诱导NSCLC细胞吉非替尼耐药

背景与目的 表皮生长因子受体(epidermal growth factor receptor, EGFR)基因突变是非小细胞肺癌(non-small cell lung cancer, NSCLC)最常见的驱动基因突变。为延长患者生存时间，NSCLC EGFR酪氨酸激酶抑制剂(tyrosine kinase inhibitors, TKIs)耐药是目前急需解决的重大难题。本研究主要探究烟草烟雾(cigaree smoke, CS)诱导NSCLC发生吉非替尼耐药的机制。方法 体外培养PC-9、A549细胞，分别经1μmol/L吉非替尼处理4 h、10%CS萃取物(CS extract, CSE)处理48 h。Western blot检测沉默蛋白3(Sirtuin 3, Sirt3)、超氧化物歧化酶2(superoxide dismutase 2, SOD2)蛋白表达，使用DCFH-DA探针检测细胞内活性氧(reactive oxygen species, ROS)水平，CCK-8试剂盒检测细胞活性，Ed U检测细胞增殖能力。Sirt3过表达质粒(OV-Sirt3)转染于PC-9和A54...     

Ringworm in animals in a farm in Assiut

Bericht über das Auftreten von Hautmykosen bei Tieren auf der Farm der Landwirtschaftlichen Fakultät der Universität Assiut. Von 70 Kälbern waren 42 pilzkrank, davon 22 durch T. mentagrophytes, 20 durch T. verrucosum. Von 180 Kühen waren 8 pilzbefalien, sämtlich durch T. mentagrophytes. Von 2 kranken Bullen wurde T. verrucosum isoliert. Auch die 3 Pferde und 2 Maultiere der Farm waren pilzinfiziert; Erreger war in diesen Fällen T. equinum. Auch ein Kalb war von T. equinum befallen. Die gleiche Pilzart wurde ferner von 3 Tierpflegern isoliert, die Pilzherde am Hals und an den Armen aufwiesen.     

Statistical challenges in preprocessing in microarray experiments in cancer

Many clinical studies incorporate genomic experiments to investigate the potential associations between high-dimensional molecular data and clinical outcome. A critical first step in the statistical analyses of these experiments is that the molecular data are preprocessed. This article provides an overview of preprocessing methods, including summary algorithms and quality control metrics for microarrays. Some of the ramifications and effects that preprocessing methods have on the statistical results are illustrated. The discussions are centered around a microarray experiment based on lung cancer tumor samples with survival as the clinical outcome of interest. The procedures that are presented focus on the array platform used in this study. However, many of these issues are more general and are applicable to other instruments for genome-wide investigation. The discussions here will provide insight into the statistical challenges in preprocessing microarrays used in clinical studies of cancer. These challenges should not be viewed as inconsequential nuisances but rather as important issues that need to be addressed so that informed conclusions can be drawn.     

Trends in zygomycosis in children

Zygomycosis, or mucormycosis, is associated with significant morbidity and mortality in both children and adults. Studies in adults have shown an increase in the incidence of zygomycosis, particularly among haemtopoietic stem cell transplant (HSCT) recipients and patients with haematologic malignancies. There is a paucity of data on the epidemiology of zygomycosis in children. We performed a retrospective analysis to describe trends in zygomycosis between 1 January 2003 and 31 December 2010. We used the Pediatric Health Information System (PHIS) database to identify paediatric patients who were diagnosed with zygomycosis during the study period. Administrative data on diagnoses, demographics, underlying conditions and clinical experiences were collected. Summary statistics were calculated and tests for trend were conducted. We identified 156 unique patients with zygomycosis. The prevalence of zygomycosis did not significantly increase over time (P=0.284). The most common underlying condition was malignancy (58%) and over half received intensive care. Voriconazole utilisation among all hospitalised children significantly increased during the period (P=0.010). Our study demonstrates that the incidence of zygomycosis is not significantly increasing. During the time period there was a significant increase in the use of voriconazole among children.     

Hemoglobin in lymphogranulomatosis in children

The peculiarities of hemoglobin content and properties in lymphogranulomatosis were studied in children by different methods. In most patients the content of fetal hemoglobin and methhemoglobin was found to increase and the rate of ferricyanide oxidation of hemoglobin also increases. Simultaneously, the rate of non-homogenicity of basic hemoglobin fractions was observed to increase on account of immunologically different components.     

Dermatomycosis in Goats in India

Summary: In an extensive survey involving 2176 goats 1.56% of goats manifested clinical lesions of ringworm infection. Animals below the age of 6 months were affected most (4.20%). The incidence of infection was higher during the winter months. T. verrucosum, T. mentagrophytes and M. gypseum were isolated from the skin scrapings of 12, 4, and 2 goats respectively. It is suggested that the infected animals could be a source of disease to human population in which the zoophilic dermatophyte invasion is characterized by severe inflammatory lesions of the skin. The zoonotic importance of different dermatophytes is stressed.
Zusammenfassung: In einer ausgedehnten Untersuchung an 2176 Ziegen wurden bei 1,56% klinische Zeichen einer Hautmykose festgestellt. Tiere, die jühger als 6 Monate alt waren, zeigten mit 4,20% am häfigsten Krankheitssymptome. In den Wintermonaten war die Erkrankungshäufigkeit am gröBten. T. vermcosum, T. mentagrophytes and M. gypseum wurden jeweils von 12,4 bzw. 2 Ziegen isoliert. Die inflzierten Tiere köinnen eine Infektionsquelle für die menschliche Bevölkerung darstellen und dort Mykosen mit stark entzündlichen Veränderungen auslösen. Die Bedeutung verschiedener Dermatophyten als Erreger von Zoonosen wird hervorgehoben.     

Sex hormones in women in rural China and in Britain

Plasma concentrations of certain hormones linked to breast cancer risk were measured in age-pooled samples from 3,250 rural Chinese women in 65 counties, and 300 British women, all aged 35-64. In age-groups 35-44, 45-54 and 55-64 respectively, mean oestradiol concentrations were 36% (P = 0.043), 90% (P less than 0.001) and 171% (P = 0.001) higher in the British than in the Chinese women, and mean testosterone concentrations were 48% (P less than 0.001), 68% (P less than 0.001) and 53% (P = 0.001) higher in the British than in the Chinese women. The difference in testosterone concentrations between the two countries appeared to be due largely to the lower average body weight in the Chinese women. Sex hormone binding globulin did not differ significantly between the two countries in age groups 35-44 and 45-54, but was 15% (P = 0.002) lower in the British than in the Chinese women at ages 55-64. Prolactin concentrations did not differ significantly between the two countries in any age group.     

Temozolomide treatment in glioma--experiences in two university hospitals in Finland

Thirty-two patients with relapsing glioma were treated with temozolomide in two university hospitals in Finland. One patient (3%) had complete response and 9 (28%) partial response, with 8 patients (25%) showing stable disease. Median progression-free survival for these 18 patients (56%) was 7 months (range 2-11+). The remaining either had progressive disease (25%) or only clinical evaluation (19%). Karnofsky score improved in 34% of patients and decreased in 3%. Symptoms were alleviated in 44% and deteriorated in 9%. Grade 3-4 toxicity was detected in 9% of the patients. Only 4% of the days in treatment were spent in hospital. An average 1.8 neuroradiological investigations, 6.9 laboratory visits, and 5.3 visits to the oncologist were made. This study confirms that temozolomide has positive effects on the outcome of often heavily pretreated glioma patients. High drug costs are compensated by prolonged home care and even the possibility to maintain working capacity.