血清lncRNA H19在非小细胞肺癌筛查及预后评估中的价值

目的分析非小细胞肺癌(NSCLC)患者血清lnc RNA H19表达水平及其筛查和预后价值。方法实时荧光定量PCR检测70例NSCLC患者和60例体检健康者血清lnc RNA H19表达水平,并分析血清H19水平与NSCLC患者临床病理参数的关系;利用ROC曲线分析H19对NSCLC的筛查效能;Kaplan-Meier法绘制不同H19水平的NSCLC患者生存曲线,Cox多因素回归模型分析NSCLC预后的独立危险因素。结果NSCLC组血清H19表达水平(1.72±0.27)明显高于健康人对照组(1.00±0.08),差异有统计学意义(t=19.911,P0.01)。ROC曲线结果表明,血清H19筛查NSCLC的AUC~(ROC)为0.864,特异性为81.7%,敏感性为74.3%。血清H19表达与TNM分期、肿瘤大小、淋巴结转移有关(P均0.05),但与年龄、性别、吸烟史、分化程度、病理分型等无关(P0.05)。H19高表达组中位生存时间为35个月,而H19低表达组中位生存时间为49个月,差异有统计学意义(Log-rankχ~2=4.874,P=0.027)。TNM分期、肿瘤大小、淋巴结转移、血清H19表达水平是影响NSCLC患者预后的独立风险因素(P均0.05)。结论NSCLC患者血清Lnc RNA H19明显升高,并且与NSCLC不良预后密切相关,有望成为NSCLC筛查和预后评估的分子标志物。     

血清外泌体源性长链非编码RNA RP N2-4在胃癌中的表达及其筛查价值

目的探讨外泌体源性长链非编码RNA RPN2-4(lncRPN2-4)在胃癌患者血清中的表达及其筛查胃癌的价值。方法收集47例胃癌患者与32例行胃镜检查者血清标本;用试剂盒法分离外泌体,并提取RNA;粒径分析和透射电镜法鉴定外泌体;qRT-PCR法检测外泌体lncRPN2-4的表达水平;电化学发光法检测血清CEA、CA19-9与CA72-4的表达水平;利用ROC曲线评价其筛查胃癌的效能。结果与胃部良性疾病组[0.91(-2.98,6.92)]相比,胃癌组血清外泌体中lncRPN2-4的表达水平[3.78(-0.06,9.12)]明显降低(U=343,P0.01);LncRPN2-4的相对表达量与肿瘤分化程度(P=0.005)和淋巴结转移(P=0.033)有关; lncRPN2-4筛查胃癌时的ROC曲线下面积(AUC~(ROC))为0.772(95%CI:0.662～0.882,P0.01),当cut-off值为1.31时,其敏感性为87.2%,特异性为59.4%。结论血清外泌体来源的lncRPN2-4在胃癌患者中低表达,可能作为胃癌筛查的肿瘤标志物。     

长链非编码RNA HOTAIR在胃癌中的表达及意义

目的:探讨长链非编码RNA HOTAIR(lnc RNA HOTAIR)在胃癌组织中的表达情况及其临床意义。方法:采用实时荧光定量PCR技术检测42例胃癌患者癌组织与癌旁组织中HOTAIR的表达水平,分析lnc RNA HOTAIR表达水平与临床病理特征之间的关系。结果:胃癌组织中lnc RNA HOTAIR表达阳性率较癌旁组织明显升高,癌组织中lnc RNA HOTAIR表达与肿瘤分化程度及临床分期密切相关。结论:胃癌组织中HOTAIR的表达明显升高,且癌组织分化越低,HOTAIR表达越高,可以将HOTAIR作为判断胃癌恶性程度的一个分子标志。     

胃癌外体（exosome）内miRNA-1的检测分析

目的检测胃癌细胞系来源外体(exosome)及其胃癌组织和血清中exosome内miRNA-1的表达水平并分析其临床意义。方法采用实时荧光定量RT-PCR分别检测胃癌细胞系来源exosome、胃癌患者组织及血清exosome内miRNA-1的表达水平,分析其与临床资料相关性,并绘制ROC曲线进行效能分析。结果 miRNA-1在人胃癌细胞系MGC-803(6.51±0.41)和SGC-7901(14.81±1.30)中的含量明显高于胃上皮细胞系GES-1(1.05±0.25),差异有统计学意义(t分别为11.26、10.38,P均0.01);而MGC-803、SGC-7901细胞培养上清exosome内miRNA-1的表达水平(49.98±11.77和28.68±4.66)显著高于GES-1来源exosome(1.00±0.02)差异有统计学意义(t分别为4.162、5.942,P均0.01);25对胃癌组织标本中有18例癌组织miRNA-1表达上调,7例表达下调,胃癌组织[1.110(0.070,4.307)]平均表达水平高于癌旁组织[1.149(1.110,2.075)],差异有统计学意义(Z=-1.897,P0.05)。miRNA-1在胃癌患者血清exosome内的表达水平[4.130(0.151,12.720)]较体检健康者血清exosome内表达水平[0.704(0.077,7.243)]明显增高(Z=-2.407,P0.05),且与胃癌的淋巴结转移相关;胃癌患者血清exosome内miRNA-1的ROC曲线下面积(AUCROC)为0.723 0,95%可信区间(CI)为0.627 0~0.818 7,cut off值为2.39,敏感性为66%,特异性为68%。结论胃癌组织及其胃癌患者血清来源exosome内富含miRNA-1,有望成为新的胃癌诊断标志物。     

长链非编码RNA在乳腺癌组织与外周血中的表达

目的探讨长链非编码RNA(lnc RNA)在乳腺癌组织与外周血中的表达。方法纳入86例乳腺癌患者和60例乳腺良性结节患者,前者收集癌组织、癌旁组织和血清标本,后者收集血清标本,实时定量PCR检测H19、HOTAIR、MALAT1和GAS5的表达水平。结果 H19、HOTAIR和MALAT1在乳腺癌组织中的表达较正常组明显升高,而GAS5在癌组织中表达下调(P<0.01)。进一步分析发现,lnc RNA表达水平与患者临床病理特征如年龄、肿瘤大小、淋巴结转移、病理类型、分期等无关。H19、HOTAIR和MALAT1在乳腺癌血清标本中的表达水平均明显高于乳腺良性结节患者(P值分别为0.011、0.004、0.010)。受试者工作特征(ROC)曲线分析表明,血清lnc RNA表达水平对乳腺癌的诊断敏感度和特异度均较满意。结论 lnc RNA在乳腺癌发展过程中具有重要作用,血清H19、HOTAIR和MALAT1水平可作为乳腺癌早期诊断的潜在标记物。     

环状RNA hsa_circ_002059在胃癌患者组织与循环血液中的表达及意义

目的建立胃癌患者癌组织、血浆和血浆外泌体中hsa_circ_002059表达的检测方法,探讨其在胃癌组织与循环血液中表达的临床应用价值。方法应用circ2Traits和starBase v2.0数据库预测胃癌相关环状RNA hsa_circ_002059的表达;实时荧光定量RT-PCR检测配对的胃癌组织与癌旁组织(87例)、胃癌患者和体检健康者血浆(52例)及外泌体(62例)样本中hsa_circ_002059表达水平的差异,并分析与临床病理参数的相关性;ROC曲线和生存曲线分析评价其在胃癌诊断与预后评估中的临床价值。结果 hsa_circ_002059具有多个胃癌相关miRNA结合位点,在胃癌组织中的表达水平显著低于癌旁组织(U=1 759.0,P0.001),且与肿瘤大小有关(χ~2=6.183,P=0.013)。组织hsa_circ_002059筛查胃癌的ROC曲线下面积(AUC~(ROC))为0.768,敏感性和特异性分别为0.67和0.92。在胃癌患者血浆中hsa_circ_002059的表达水平明显高于体检健康者(U=1 011.5,P=0.027),且与淋巴结转移有关(χ~2=12.231,P=0.005)。血浆hsa_circ_002059筛查胃癌的AUC~(ROC)为0.626,敏感性和特异性分别为0.65和0.64。此外,hsa_circ_002059高表达患者的平均生存时间显著低于低表达者(χ~2=4.594,P=0.032),在胃癌患者血浆外泌体中hsa_circ_002059的表达水平与体检健康者之间的差异无统计学意义(U=1 746.0,P=0.379)。结论 hsa_circ_002059表达与胃癌进展有关,其在组织和血浆中表达水平的检测有望用于胃癌辅助诊断与预后评估。     

血清长链非编码RNA CRNDE在鼻咽癌中的表达及临床应用价值

目的探讨血清长链非编码RNA(long non coding RNA,LncRNA)结直肠肿瘤差异表达基因(colon rectal neoplasia differentially expressed,CRNDE)在鼻咽癌患者血清及组织中的表达水平及其临床应用价值。方法采用实时荧光定量PCR(qRT-PCR)检测鼻咽癌患者(n=78)和健康人群(n=78)血清和组织中CRNDE的表达水平,并分析CRNDE与鼻咽癌患者临床病理参数的相关性;采用CCK8增殖试验检测鼻咽癌细胞系CNE-2Z(WT)和CNE-2Z(Cas9-CRNDE)的细胞增殖能力的差异。ROC曲线分析CRNDE在早期鼻咽癌患者筛查中的临床应用价值。结果鼻咽癌患者血清CRNDE的表达水平明显高于健康人群(5.27±0.89 vs 1.01±0.12,P0.001);且CRNDE表达与T分期、N分期、TNM分期、淋巴结转移、侵犯颈动脉鞘、侵犯颅底相关(P均0.05);鼻咽癌患者癌组织中CRNDE的表达水平亦明显高于癌旁组织(4.56±0.38 vs 1.82±0.21,P0.05);经CRNDE敲除后,与阴性对照组细胞相比,鼻咽癌系细胞的增殖速率明显降低(2.42±0.13 vs 5.81±0.48,P0.05);血清CRNDE筛查鼻咽癌的ROC曲线下面积(AUC~(ROC))为0.835(95%CI:0.598～0.984),当cut-off值为11.25时,敏感性和特异性分别为93.1%、81.7%。结论鼻咽癌患者血清和组织中CRNDE的表达明显升高,有助于鼻咽癌早期筛查及病情评估。     

长链非编码RNA LincROR在卵巢癌筛查中的应用

目的分析卵巢癌患者血浆和组织中LincROR的表达,探讨LincROR在卵巢癌筛查中的价值。方法收集30例健康女性、56例卵巢囊肿、23例子宫内膜异位症、38例子宫内膜癌、35例宫颈癌、42例卵巢癌、21例卵巢癌术后及26例卵巢癌化疗后患者的血浆样本,荧光定量PCR分析LincROR在各组样本中的表达水平,结合临床数据评估LincROR在临床妇科常见疾病中的筛查效能。结果 LincROR在卵巢癌患者血浆中的表达水平(2.90±4.42)明显高于健康人(0.23±0.28)和卵巢良性囊肿患者(0.62±0.55),差异有统计学意义(P均0.01)。ROC曲线分析结果表明,血浆LincROR在卵巢癌筛查中的价值优于CA125、CA199、CA153、AFP和CEA等指标, LincROR和CA125联合筛查卵巢癌的敏感性为89.7%,特异性为86.7%(AUC~(ROC):0.918, 95%CI:0.817～0.973)。此外,卵巢癌术后患者血浆中LincROR的表达水平明显低于未经任何治疗的卵巢癌患者(0.50±1.72 vs 2.90±4.42,P0.01)。ROC曲线分析表明,血浆LincROR是比CA125更敏感的卵巢癌化疗疗效的评价指标(AUC~(ROC):0.866 vs 0.738)。结论 LincROR有望成为卵巢癌筛查的生物学标志物,联合CA125可提高筛查的敏感性和特异性,同时在评估卵巢癌化疗疗效中具有潜在的临床价值。     

胃癌患者血清外泌体中DANCR表达及其临床意义

目的检测胃癌患者血清外泌体(exosome)中抗分化非编码RNA(DANCR)的表达水平并分析其临床应用价值。方法采用实时荧光定量RT-PCR检测胃癌细胞培养上清和胃癌患者血清exosome中DANCR表达水平,分析其与临床病理资料的相关性,绘制ROC曲线评价其诊断效能。结果 DANCR在人胃癌细胞系HGC-27中表达水平(4.43±0.37)明显高于胃黏膜上皮细胞(1.01±0.01),差异有统计学意义(P0.05)。HGC-27细胞培养上清exosome中DANCR表达水平(3.06±0.14)明显高于GES-1细胞(1.01±0.20),差异有统计学意义(P0.05)。DANCR在胃癌患者血清exosome中的平均含量[5.060(1.380,22.785)]较胃良性疾病患者[0.535(0.340,1.380)]和体检健康者[1.200(0.605,1.655)]明显升高(Z分别为-3.409,-4.229,P均0.01),且与肿瘤大小、TNM分期以及淋巴结转移相关。胃良性疾病患者与体检健康者相比,血清exosome中DANCR含量差异无统计学意义(Z=-1.308,P0.05)。胃癌患者血清exosome中DANCR的ROC曲线下面积(AUCROC)为0.777,95%可信区间(CI)为0.678~0.876,cut-off值2.50,敏感性为68.6%,特异性为84.4%。结论胃癌患者血清exosome中DANCR表达水平升高,有望成为胃癌诊断的新指标。     

血清长链非编码RNA LSINCT5在乳腺癌中的表达与临床意义

目的探讨长链非编码RNA(long-noncoding RNA,lnc RNA)LSINCT5在乳腺癌患者血清中的表达水平及其临床应用价值。方法分别搜集90例乳腺癌患者、88例乳腺良性疾病患者及94例体检健康者血清标本,以及35例乳腺癌患者术前和术后血清标本;荧光定量PCR(q RT-PCR)法检测各组血清中LSINCT5的表达水平,并分析其与临床病理参数的相关性;采用电化学发光免疫测定法(ECLI)检测各组血清中CA153、CEA的表达水平,并进行多元Logistic回归分析;绘制ROC曲线评估血清LSINCT5对乳腺癌的诊断效能。结果乳腺癌组血清LSINCT5表达水平[1.45(0.76,3.16)]明显高于良性疾病组[1.08(0.66,1.45),H=3.188,P=0.004]和健康人对照组[1.21(0.80,1.44),H=2.626,P=0.026];术后血清中LSINCT5表达水平[0.85(0.49,1.31)]较术前明显降低[1.10(0.67,2.23),U=0.764,P=0.005];此外,血清LSINCT5水平与乳腺癌患者TNM分期、淋巴结转移、PR、和Ki-67有关(P0.05),而与年龄、病理类型、ER和Her2无关(P0.05);血清LSINCT5单独诊断乳腺癌的ROC曲线下面积(AUC~(ROC))为0.60,敏感性和特异性分别为39.7%和100%,均高于传统标志物CA153(AUC~(ROC)=0.59,25.0%,95.6%)和CEA(AUC~(ROC)=0.54,33.8%,82.4%),且3项指标联合后其诊断效能(AUC~(ROC)=0.66,45.6%,100%)高于各项单独检测。结论乳腺癌患者血清中LSINCT5高表达,可能作为乳腺癌诊断的一个潜在的生物学标志物。     

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.