Sorafenib对前列腺癌DU145细胞的抑制作用的研究

目的观察索拉非尼（Sorafenib）对雄激素非依赖性前列腺癌Du145细胞的抑制作用。方法用不同浓度Sorafenib处理前列腺癌DU145细胞24、48和72h后,MTT法检测Sorafenib对DUl45细胞的抑制作用,流式细胞仪检测细胞凋亡变化,Westernblot检测不同浓度Sorafenib处理72h后DU145细胞内ERK和Bcl-2的表达。结果Sorafenib能显著抑制DU145细胞的体外生长,呈时间与剂量依赖性。DUl45细胞凋亡率随着Sorafenib剂量的增加而增大,具有良好的量效关系（P〈0．01）;Sorafenib处理DU145细胞72h后,ERK和Bcl-2蛋白的表达明显下调（P〈0．01）。结论Sorafenib抑制DU145细胞增殖、诱导细胞凋亡,可显著抑制雄激素非依赖性前列腺癌细胞的体外生长。     

EB1089对前列腺癌DU145细胞生长和凋亡的影响

目的研究EB1089对前列腺癌DU145细胞增殖、凋亡的影响,并探讨其机制。方法体外培养DU145细胞,以1、10、50、100 nmol/L的EB1089分别作用24、48、72 h,MTT法和流式细胞技术检测EB1089对DU145细胞的增殖和凋亡,Western blot检测各浓度的EB1089作用48 h后,DU145细胞Gli蛋白的表达变化。结果 EB1089可以抑制DU145细胞的增殖,诱导DU145细胞凋亡。不同浓度EB1089作用48 h后,DU145细胞Gli蛋白表达下降。结论 EB1089可能通过Hedgehog信号通路抑制DU145细胞增殖、诱导其凋亡。     

靶向血管内皮生长因子siRNA对K562细胞凋亡及survivin表达的影响

目的 应用腺病毒介导的血管内皮生长因子(VEGF) siRNA沉默K562细胞中VEGF的表达,观察其对K562细胞凋亡及凋亡抑制基因survivin表达的影响.方法 应用成功构建的携带特异性VEGF siRNA的重组腺病毒(Ad5-VEGF siRNA)感染K562细胞,实验分为3组:实验组(K562/Ad5-VEGF siRNA组)、空载体组(K562/Ad5组)、对照组(K562组).通过RT-PCR法检测细胞内VEGF及survivin mRNA的表达,ELISA法检测细胞培养上清中VEGF蛋白表达,Western blot法检测细胞survivin蛋白表达,流式细胞术检测细胞凋亡.结果 实验组细胞VEGF及survivin mRNA表达水平较对照组均有明显降低(P＜0.01),且细胞培养上清中VEGF蛋白表达水平[(1121±15)pg/ml]低于空载体组[(1290±28)pg/ml]和对照组[(1303±28)pg/ml](P＜0.01).Western blot法检测结果显示,实验组survivin蛋白表达水平(0.26 ± 0.11)较对照组(0.74±0.10)也显著降低(P＜0.01).实验组细胞凋亡率与对照组比较明显增加(P＜0.01).结论 应用RNA干扰沉默K562细胞中VEGF基因表达后,survivin的表达也随之降低,同时细胞凋亡率相应增加.VEGF可诱导K562细胞中survivin基因表达,可能是survivin基因表达的上游调控因子.     

茶多酚对前列腺癌PC-3M细胞增殖与凋亡的影响

目的探讨茶多酚对前列腺癌PC-3M细胞增殖及凋亡的影响。方法设立对照组与实验组,采用MTT比色法检测TP对PC-3M细胞增殖的影响;吖啶橙/溴化乙锭染色法观察诱导细胞凋亡情况;流式细胞术分析细胞周期;RT-PCR和Western blot检测Caspase-3基因的转录与表达。结果与对照组比较,MTT法示实验组对PC-3M细胞的增殖抑制作用增强;细胞染色法示实验组凋亡率增高;流式细胞术显示细胞阻滞于G1期,增殖指数降低;RT-PCR与Western blot示Caspase-3基因转录与翻译上调,上述结果在一定范围内呈时间、剂量依赖性(P0.05)。结论茶多酚对人前列腺癌PC-3M细胞具有增殖抑制、促进凋亡作用,与上调caspase-3基因的转录与表达有关。     

低频超声联合微泡增强脂质体介导Rac1-shRNA质粒转染前列腺癌细胞及对其生物学行为的影响

目的研究低频超声联合微泡增强脂质体介导的Rac1-shRNA质粒转染前列腺癌PC3、DU145细胞,并探索其对前列腺癌细胞凋亡、侵袭能力的影响。方法基础情况下Westernblot检测RWPE-1细胞、PC3细胞和DU145细胞的Rac 1蛋白表达。根据不同处理方式将细胞分为四组:PC3细胞对照组(A组)、超声加微泡联合脂质体转染PC3细胞组(B组)、DU145细胞对照组(C组)及超声加微泡联合脂质体转染DU145细胞组(D组)。通过蛋白质体外结合实验技术检测各组Rac1蛋白的活性及表达;流式细胞仪检测各组细胞早期凋亡情况;细胞侵袭实验检测各组细胞侵袭能力。结果基础情况下前列腺癌PC3、DU145细胞Rac1蛋白表达均较正常前列腺细胞RWPE-1增加,差异均有统计学意义(均P0.01);低频超声携微泡介导Rac-1shRNA质粒转染细胞后,B组Rac1蛋白表达量较A组表达减少,D组表达量较C组减少,差异均有统计学意义(均P0.01);早期细胞凋亡检测显示,B组高于A组,D组高于C组,差异均有统计学意义(均P0.01);同期细胞侵袭力实验显示,B组穿膜细胞数少于A组,D组的穿膜细胞数少于C组,差异均有统计学意义(均P0.01)。结论低频超声联合微泡可促进脂质体介导的Rac1-shRNA质粒转染人雄激素非依赖型前列腺癌细胞;通过抑制PC3、DU145细胞中Rac1蛋白表达可以促进癌细胞的早期细胞凋亡,同时可抑制癌细胞的侵袭能力。     

熊果酸对人前列腺癌DU145细胞增殖及凋亡的影响

【目的】探讨熊果酸对人前列腺癌DU145细胞增殖及凋亡的影响。【方法】常规培养前列腺癌细胞系DU145,采用不同浓度熊果酸干预48 h ,细胞增殖与毒性分析（CCK‐8）法检测DU145细胞增殖;流式细胞术检测DU145细胞的凋亡率;Western blot检测DU145细胞p‐Akt及bcl‐2蛋白表达;实时定量聚合酶链式反应（qRT‐PCR）检测 PTEN mRNA 表达。【结果】细胞增殖抑制试验显示熊果酸呈浓度依赖性抑制DU145细胞生长;0、10、20μmol／L药物处理细胞48 h后其早期凋亡率分别为（2．15±0．24）％、（13．52±1．83）％及（16．69±2．56）％,晚期凋亡率分别为（3．28±0．53）％、（18．47±2．64）％及（23．70±1．99）％,差异均有统计学意义（ P ＜0．05）;与空白对照组相比,熊果酸作用的DU145细胞p‐Akt及bcl‐2蛋白表达水平均降低（均 P ＜0．05）,PTEN mRNA表达水平上升（ P ＜0．05）。【结论】熊果酸能够浓度依赖地抑制DU145细胞增殖,其机制可能是通过抑制PT EN／P13K／Akt信号转导通路来实现的。     

Manumycin对白血病细胞生长及survivin基因表达的影响

目的:观察不同浓度的Manumycin对白血病K562细胞生长及survivin基因表达的影响,初步探讨其诱导细胞凋亡的分子机制.方法:体外培养K562细胞,采用不同浓度的Manumycin(0、0.5、1、2、4、8 μmol/L)掺入细胞,作用48 h后采用MTT法检测细胞增殖抑制率,流式细胞术检测凋亡率,RT-PCR检测细胞中survivin mRNA表达.结果:MTT法显示,1～8 μmol/L组对K562细胞具有明显的增殖抑制作用.流式细胞术结果显示,1～8μmol/L组均可诱导细胞凋亡(P＜0.01).RT-PCR结果显示1～8 μmol/L组细胞的survivin mRNA水平明显低于空白对照组(P＜0.01).结论:Manumycin可有效抑制K562细胞的增殖,诱导其凋亡,分子机制可能与下调survivin的表达有关.     

大黄素对胃癌细胞生长及Bcl-2表达的调控

目的:研究大黄素对人胃腺癌SGC-7901细胞生长及凋亡的影响及对Bcl-2基因表达水平的改变,探讨其诱导肿瘤细胞凋亡的可能机制.方法:将不同浓度的大黄素作用于体外培养的人胃腺癌细胞,MTT法观察细胞增殖率的改变;流式细胞仪检测细胞凋亡;细胞免疫组化法检测Bcl-2蛋白表达.结果:各实验组均抑制胃癌细胞的生长,细胞增殖率随大黄素浓度的升高和作用时间的延长而下降,与对照组相比差异有统计学意义(P ＜ 0.01);各实验组作用于胃癌细胞48 h后,凋亡细胞可以检出且与对照组相比差异有统计学意义(P ＜ 0.01);各实验组可不同程度地下调胃癌细胞Bcl-2蛋白表达水平,与对照组相比差异有统计学意义(P ＜ 0.01),并呈浓度相关性.结论:大黄素体外可以抑制人胃腺癌SGC-7901细胞的增殖及诱导细胞的凋亡;大黄素诱导SGC-7901细胞凋亡可能与其下调Bcl-2蛋白表达有关.     

2-甲氧基雌二醇对慢性髓系白血病K562细胞Caspase-3及Survivin表达的影响

本研究探讨2-甲氧基雌二醇（2-ME）对慢性髓系白血病（CML）K562细胞caspase-3和survivin表达的影响。实验分3组：对照组,培养基中不含2-ME;实验组,分别用1、2、4、8及16μmol／L的2-ME处理K562细胞;阴性对照组,培养液中以不合RNase的灭菌蒸馏水代替K562细胞。应用TUNEL、流式细胞术（FCM）、半定量RT—PCR分别检测K562细胞凋亡率、caspase-3及survivin蛋白及其基因表达水平。结果表明：2-ME在一定的浓度范围内呈剂量依赖形式诱导K562细胞凋亡,与对照组比较差异均有统计学意义（P均〈0．01）,且TUNEL和FCM方法检测的K562细胞凋亡率呈正相关（γ=0．845,P=0．034）。随着2-ME浓度增加,caspase-3蛋白表达量逐渐增加,而survivin蛋白表达量逐渐降低,两者与对照组比较差异均有统计学意义（P均〈0．05）;且caspase-3与survivin蛋白表达量之间呈负相关（γ=-0．956,P=0．001）。随着2-ME浓度增加,caspase-3基因表达量逐渐增加,而survivin基因表达量逐渐降低,两者与对照组比较差异均有统计学意义（p均〈0．01）,且caspase-3与survivin基因表达量之间呈负相关（γ=0．966,P=0．001）。结论：2-ME能够以剂量依赖形式诱导K562细胞凋亡,显示了其对CML有潜在的治疗价值。     

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.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

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.     

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.