ECT2通过调控EGFR介导的上皮间质转化促进胰腺癌转移

目的研究上皮细胞转化因子2(ECT2)在胰腺癌转移的作用及其可能的诱导表皮生长因子受体(EGFR相关的上皮-间充质转化(EMT)机制。方法使用免疫组织化学和Western blotting方法检测ECT2在胰腺癌组织及细胞系中的表达情况。在胰腺癌细胞中降低ECT2的表达,通过细胞划痕实验测定胰腺癌的细胞迁移变化,使用Transwell实验检测胰腺癌细胞的侵袭力。通过Western blotting方法测定EMT标志物和EGFR的表达变化。结果 ECT2在胰腺癌组织和细胞中显著上调,并提示不良预后。ECT2调控胰腺癌细胞的侵袭和迁移。ECT2沉默下调EGFR表达同时抑制EMT。结论 ECT2可能通过EGFR信号通路相关的EMT促进胰腺癌细胞的侵袭性。     

辛伐他汀通过MEK/ERK信号转导通路上调突触素表达

目的研究辛伐他汀(SV)对体外培养大鼠大脑皮层神经元突触素(SYP)的影响及信号转导机制。方法从新生Sprague-Dawley大鼠大脑皮层分离和培养神经元,4 d后分为对照组、SV处理组(2、4、8μmol/L作用48 h)、PD98059处理组和SV+PD98059处理组(先加入10μmol/L阻断剂PD98059作用30 min,再加入10μmol/L SV作用48 h)。应用免疫荧光检测法检测SYP表达,Western blot检测磷酸化丝裂原细胞外激酶(p-MEK)、磷酸化细胞外调节蛋白激酶1(p-ERK1)和磷酸化细胞外调节蛋白激酶2(p-ERK2)水平。结果免疫荧光检测显示,4μmol/L SV可明显增强SYP免疫反应性。Western blot检测显示,8μmol/L SV可显著增高SYP、p-MEK、p-ERK1和p-ERK2水平。应用PD98059可显著降低SV引起的SYP、p-MEK、p-ERK1和p-ERK2水平上调。结论SV可通过激活MEK/ERK信号转导通路上调SYP表达。     

人肺腺癌厄洛替尼耐药细胞系PC-9/ER的建立及其特性

目的:构建厄洛替尼耐药人肺腺癌细胞模型PC-9/ER,观察单用人表皮生长因子受体(epithelial growth factor receptor,EGFR)酪氨酸激酶抑制剂厄洛替尼或联合胰岛素样生长因子受体1(insulinlike growth factor receptor 1,IGFIR)酪氨酸激酶抑制剂苦鬼臼毒(picropodophyllotoxin,PPP)作用于该细胞后,该细胞对厄洛替尼耐药性的影响,并探讨耐药相关机制。方法:选择人肺腺癌细胞株PC-9,采用逐步递增厄洛替尼浓度的方法体外诱导构建耐药株PC-9/ER,CCK-8法检测耐药指数;细胞计数法绘制PC-9和PC-9/ER的生长曲线,并计算出两细胞系的倍增时间;流式细胞术检测两细胞系的细胞周期;Western Blot法检测p-EGFR及p-IGFIR的表达水平,并进一步检测厄洛替尼及PPP单独或联合作用于PC-9/ER后,各组Akt,ERK,p-Akt及p-ERK的表达水平。结果:PC-9/ER细胞株的耐药指数是72.3。细胞生长曲线显示,PC-9/ER细胞生长较亲代细胞慢,PC-9与PC-9/ER细胞的倍增时间分别为32.9及36.9 h(P=0.003)。与PC-9相比,PC-9/ER细胞的G1期细胞增多(P=0.001),而S期细胞则明显下降(P:0.015)。Western Blot结果表明,PC-9/ER细胞中p-IGF1R表达比亲代细胞明显增多(P=0.016),而p-EGFR无明显变化(P=0.152)。在亲代及耐药细胞系,厄洛替尼联合PPP组均较其他组更能显著的抑制细胞增殖(P<0.05);且Western Blot法表明联合用药组EGFR下游磷酸化的Akt、ERK的表达水平明显减少。结论:成功构建了人肺腺癌厄洛替尼耐药细胞株PC-9/ER,IGF1R通路可能与肺腺癌EGFR-TKI获得性耐药有关。     

靶向阻断表皮生长因子受体信号通路影响鼻咽癌细胞生长的研究

目的:利用Erlotinib(厄洛替尼,EGFR-酪氨酸激酶拮抗剂,商品名tarceva)阻断表皮生长因子受体(EGFR)介导的信号通路,观察其对鼻咽癌细胞株CNE-2生长的影响,并探索其机制.方法:用不同浓度Erlotinib处理鼻咽癌细胞株CNE-2,采用Cell Count Kit-8(CCK-8)法检测细胞生长抑制率,流式细胞术进行细胞周期、凋亡率的分析,Western bloting检测相关蛋白表达.结果:(1)Erlotinib呈剂量依赖性抑制CNE-2细胞生长;(2)处理组细胞G1期细胞比例和凋亡率明显高于对照组(P＜0.05);(3)与对照组相比,处理组出现明显的p-EGFR表达下调.结论:Erlotinib通过抑制EGFR的磷酸化阻断EGFR信号通路,增加G1期阻滞,从而抑制鼻咽癌细胞生长.     

非小细胞肺癌组织表皮生长因子受体磷酸化水平与患者临床疗效的关系

目的:探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中表皮生长因子受体(epidermal growth factor receptor,EGFR)及其磷酸化水平与患者临床生物学特征、临床治疗疗效间的关系。方法 :收集NSCLC患者75例,采用免疫组化检测其肿瘤组织中EGFR和磷酸化EGFR(p-EGFR)的表达水平,分析其与肺癌生物学特征及疗效间的关系。结果:本研究NSCLC患者的肿瘤组织中EGFR表达阳性率达78.67%,p-EGFR阳性率为56.00%,均明显高于癌旁肺组织;EGFR表达水平与肿瘤细胞的分化程度有关,而p-EGFR表达水平与肿瘤细胞的分化程度、淋巴结转移及临床分期均密切相关,低分化肿瘤细胞、发生肿瘤转移者的肿瘤组织、晚期肿瘤组织中的p-EGFR表达水平均明显升高。同时,p-EGFR水平与NSCLC患者临床治疗的有效率密切相关,p-EGFR表达阴性的患者其治疗有效率明显高于阳性者。结论:EGFR的磷酸化形式在判断NSCLC疾病进展及临床疗效时可能更有实用意义,在评估NSCLC患者的临床疗效及预后判断中具有一定价值。     

缺氧诱导因子-1α和磷酸化细胞外信号调节激酶在胃癌中的表达和意义

目的研究胃癌组织中缺氧诱导因子-1α（HIF-1α）和磷酸化细胞外信号调节激酶（pERK）的表达特点和意义。方法常规石蜡包埋行HIF-1α和p-ERK免疫组织化学染色。结果HIF-1α和p-ERK在Ⅲ／Ⅳ期、低分化、癌细胞浸润浆膜层和淋巴结转移的胃癌组织中的阳性表达明显高于Ⅰ／Ⅱ期、高分化、癌细胞未浸润至浆膜层和淋巴结未转移的胃癌组织,组间比较差异均有显著统计学意义（P〈0．01）;HIF-1α和p-ERK在胃癌中表达密切相关（r=0．467,P〈0．05）。结论HIF-1α和p-ERK表达与胃癌发生发展生物学行为和预后可能有关,可作为重要的生物学标记物。     

靶向联合阻断EGFR信号通路影响鼻咽癌细胞生长的研究

目的 本研究利用tarceva(EGFR-酪氨酸激酶拮抗剂)阻断EGFR介导的信号通路,观察其对鼻咽癌细胞株CNE-２生长的影响,并探索其机理。方法　用不同浓度的 tarceva处理鼻咽癌细胞株CNE-２,采用CCK-8法检测细胞生长抑制率,流式细胞术进行细胞周期、凋亡率的分析,western bloting检测相关蛋白表达。结果　（1）tarceva呈剂量依赖性抑制CNE-2细胞生长;( 2 )处理组细胞G1期细胞比例和凋亡率明显高于对照组（P<0.05）;（3）与对照理组相比,处理组出现明显的p-EGFR表达下调。结论 tarceva通过抑制EGFR的磷酸化阻断EGFR信号通路,增加G1期阻滞,从而抑制鼻咽癌细胞生长。     

RNA结合蛋白6对喉癌发生发展的调节作用及机制研究

目的探讨RNA结合蛋白6(RBM6)对喉癌发生发展的调节作用及机制。方法收集15例原位喉癌患者在术中的喉癌组织及癌旁组织,采用实时荧光定量PCR(RT-qPCR)和Western Blot检测RBM6、表皮生长因子受体(EGFR)、细胞外信号调节激酶(ERK)和磷酸化ERK(p-ERK)的表达。将携带RBM6基因(Lv-RBM6,RBM6组)或空质粒(Lv-Ctrl,对照组)的重组慢病毒载体转染入人喉癌细胞系LCC构建RBM6过表达喉癌细胞及对照细胞,采用CCK-8法和Transwell法分别检测细胞增殖、迁移和侵袭,并检测MMP-2、MMP-9、EGFR、ERK和p-ERK的表达。将稳定表达Lv-RBM6或Lv-Ctrl的LCC细胞注射于裸小鼠右侧颈背部皮下组织,建立Lv-RBM6过表达人喉癌裸小鼠移植瘤模型,每2天计算移植瘤体积。结果与癌旁组织对比,RBM6在喉癌组织中的表达显著降低(P 0. 05),EGFR和p-ERK的表达显著增加(P 0. 05),ERK无显著变化。RBM6在3/4期患者喉癌组织中的表达显著低于其在1/2期患者中的表达(P 0. 05)。体外实验显示,与对照组相比,RBM6组细胞在12 h、24 h、48 h、72 h的细胞增殖活性均显著降低(P 0. 05),迁移细胞数及侵袭细胞数均显著降低(P 0. 05),MMP-2、MMP-9、EGFR和p-ERK的表达显著降低(P 0. 05),ERK无显著变化。异种移植瘤实验显示,与Lv-Ctrl组相比,Lv-RBM6组裸小鼠接种后12 d、14d、16 d、20 d、24 d、28 d肿瘤体积显著降低(P 0. 05),EGFR和p-ERK的表达显著降低(P 0. 05),ERK无显著变化。结论 RBM6可抑制喉癌的生长和进展,作为肿瘤抑制因子抑制喉癌细胞的增殖、迁移和侵袭并诱导其凋亡,其机制可能与抑制EGFR相关信号传导途径的活性有关。     

联合抑制Notch2和MEK/ERK通路对胃癌细胞增殖的影响

目的探讨Notch2和MEK/ERK信号通路在胃癌细胞SGC-7901中是否存在交叉作用。方法采用体外化学合成的特异性针对Notch2的siRNA(Notch2siRNA)和丝裂原激活蛋白激酶(MEK)/细胞外信号调节激酶(ERK)信号通路的抑制剂PD98059,分别单独和联合处理体外培养的胃癌SGC-7901细胞,以转染阴性对照siRNA(control siRNA)细胞作为siRNA对照组,并设不给予任何转染的空白对照组。免疫印迹(Western Blotting)法检测磷酸化ERK1/2(p-ERK)1/2和Notch2蛋白的表达水平。比色法(MTT)检测癌细胞增殖抑制率。结果Notch2siRNA能降低蛋白Notch2的表达水平,并抑制癌细胞增殖[(38.26±1.82)%],而p-ERK的表达水平则较对照组增加。PD98059能降低p-ERK的表达水平,并抑制癌细胞的增殖[(30.05±3.16)%],Notch2水平则无明显变化,联合应用Notch2siRNA和PD98059能明显降低p-ERK和Notch2蛋白的表达水平,与Notch2siRNA或PD98059单独应用比较,显著抑制癌细胞增殖率,差异有统计学意义[(72.55±5.30)%,P0.01]。结论特异性抑制Notch2信号通路,且抑制MEK/ERK通路可进一步增强抑制Notch2通路的抗肿瘤增殖效果,提示MEK/ERK和Notch2 2条信号通路在胃癌SGC-7901细胞中存在交叉作用。     

西妥昔对脂多糖诱导的小胶质细胞活化和迁移的影响

目的:研究表皮生长因子受体(EGFR)的特异性单克隆抗体西妥昔(Cetuximab)对小胶质细胞活化及迁移的影响.方法:细菌脂多糖(LPS)诱导活化原代培养的大鼠脊髓小胶质细胞,给予西妥昔干预,免疫荧光法观察小胶质细胞形态及磷酸化EGFR表达水平的变化,Transwell法检测细胞的迁移.结果:与对照组相比,LPS诱导...     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.     

Wie zuverlässig sind Angaben zu nichtentzündlichen ausgedehnten Schmerzen?

ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.