Real time RT-PCR检测肝癌患者外周血AFP mRNA的表达水平

目的:了解肝细胞癌患者外周血AFP mRNA的表达水平。方法;采用real time RT-PCR技术,定量检测肝细胞癌56例、慢性活动性肝炎17例、肝硬化21例和健康志愿者20例外周血AFP mRNA的表达水平。结果:肝癌56例外周血AFP mRNA的平均表达水平为34644.6±94905.4copies/μgRNA,其中29例为阳性;慢性活动性肝炎17例、肝硬化21例和健康志愿者20例外周血均未检测到AFP mRNA的表达。结论:AFP mRNA可作为血循环中存在肝癌细胞的标志物,定量检测外周血AFP mRNA是预测肝癌发生远处转移的一个可靠手段。     

RT-PCR检测外周血CEA和CK20 mRNA对结肠癌肿瘤微转移的预测价值

目的 检测结肠癌患者外周血CEA和CK20mRNA的表达水平及评估其对肿瘤转移的预测价值.方法 分别采用RT-PCR和传统定性PCR技术,检测结肠癌患者(85例)和健康志愿者(45例)外周血CEA和CK20mRNA的表达水平.结果 85例结肠癌患者外周血CEA和CK20mRNA的水平分别为(920±104)copies/ml和(532±91)copies/ml,健康志愿者未检测到CEA和CK20 mRNA表达;CEA和CK20 mRNA水平与Dukes分期、细胞分化程度相关,Logistic回归分析得随Dukes分期进展,肿瘤转移率逐渐增加(与A期相比OR值分别为1.087、1.983、2.876);RT-PCR检测灵敏度明显高于定性PCR.结论 RT-PCR检测结肠癌患者外周血CEA和CK20 mRNA表达水平灵敏度高,CEA和CK20 mRNA具有较好的肿瘤微转移预测价值.     

肺癌患者外周血循环细胞CEA-mRNA RT-PCR检测的意义

目的　了解肺癌患者外周血CEA -mRNA的水平。方法　采用实时荧光RT- PCR技术,检测肺癌患者78例,肺部良性疾病(BLD)患者70例,健康志愿者30例外周血CEA- mRNA的水平。结果　78例肺癌患者外周血CEA- mRNA的平均水平为(10 2 38. 4±185 2 6 . 3)copies/ml,其中4 3例阳性、阳性率为5 5 . 1%。BLD患者外周血CEA mRNA平均水平为(82. 7 4±6 0. 3 7)copies/ml,其中阳性2例,阳性率为6 . 7%。健康对照未检测到CEA -mRNA。CEA -mRNA水平与肿瘤病理分期和分化程度相关;CEA -mRNA阳性组患者血清CEA蛋白的阳性率与CEA mRNA阴性组相比差异无显著性(P>0 0 5 )。结论　外周血CEA mRNA可作为肺癌辅助诊断指标,定量检测外周血CEA- mRNA可作为肺癌微转移的一个监测指标。     

荧光定量PCR检测大肠癌患者外周血CK19mRNA、CK20mRNA和CEAmRNA的意义

目的探讨荧光定量PCR技术检测大肠癌患者外周血CK19mRNA、CK20mRNA和CEAmRNA的临床意义。方法采用荧光定量PCR技术,检测大肠癌患者76例,健康志愿者30例外周血CK19mRNA、CK20mRNA和CEAmRNA水平。结果76例大肠癌患者外周血CK19mRNA的水平为5 962.5±19 926.1 copies/ml,其中38例阳性,阳性率为50.0%;CK20mRNA的水平为6 244.3±15 113.1 copies/ml,其中28例阳性,阳性率为36.8%;CEAmRNA的水平为7 377.0±26 281.5 copies/ml,其中36例阳性,阳性率为47.4%。对照组30例未见表达。单项检测阳性率较低,联合其中二项检测阳性率较高,联合三项阳性率可达68.4%。结论外周血CK19mRNA、CK20mRNA和CEAmRNA可作为大肠癌微转移辅助诊断指标,联合二至三项指标有较高的检出率,定量检测有助于大肠癌微转移的监测。     

荧光定量PCR法检测肿瘤慢性贫血患者外周血淋巴细胞铁调素mRNA

目的建立检测肿瘤慢性贫血患者铁调素(hepcidin)mRNA表达水平的荧光定量PCR法,初步观察肿瘤慢性贫血患者淋巴细胞铁调素mRNA水平。方法建立荧光定量PCR检测铁调素mRNA的标准曲线,评价其灵敏度、特异性及重复性;检测50例体检健康者、49例肿瘤慢性贫血患者外周血淋巴细胞铁调素mRNA水平,同时检测血清白细胞介素6(IL-6)及血清铁(SI)水平。结果荧光定量PCR检测铁调素mRNA的灵敏度为10 copies/μL,线性范围为101~107copies/μL;熔解曲线示扩增峰单一;检测3份低、中、高浓度的阳性质粒批内CV分别为1.83%、1.27%、1.39%,批间CV分别为6.13%、7.48%、6.87%。肿瘤慢性贫血组铁调素mRNA水平为[10.51(8.02~16.31)]×10-3copies/cell,高于健康人对照组[2.57(2.20~3.20)]×10-3copies/cell,P0.01;外周血铁调素mRNA水平与IL-6水平呈正相关(r=0.92,P0.01),与SI呈负相关(r=-0.83,P0.01)。结论建立了定量检测外周血淋巴细胞铁调素mRNA的荧光定量PCR法,肿瘤慢性贫血患者铁调素mRNA水平高于健康人,可能与炎症反应有关。     

肺癌患者外周血微转移检测的意义

目的研究肺癌患者外周血中CEAmRNA、CK19mRNA和CK20mRNA的表达及联合检测的临床意义。方法采用荧光定量PCR技术,检测48例肺癌患者、8例肺良性病变和30名健康志愿者外周血CEAmRNA、CK19mRNA和CK20mRNA水平。结果48例肺癌患者外周血CEAmRNA的水平为(16264±28765)copies/ml,其中26例阳性,阳性率为54.2%;CK19mRNA的水平为(14891±27244)copies/ml,其中27例阳性,阳性率为56.3%;CK20mRNA的水平为(10924±21678)copies/ml,其中20例阳性,阳性率为41.7%。8例肺良性病变中CK20mRNA阳性1例(12.5%),CEAmRNA和CK19mRNA均为阴性,健康志愿者30例未见表达。单项检测敏感度和准确性较低,联合其中二项检测敏感度和准确性较高,联合三项敏感度达77.1%、准确性可达87.2%。CEAmRNA、CK19mRNA和CK20mRNA水平和阳性率与肺癌类型和不同临床分期有关。结论CEAmRNA、CK19mRNA和CK20mRNA可作为标志物检测肺癌患者外周血微转移,联合二至三项指标有较高的检出率,定量检测有助于肺癌微转移的监测。     

胃癌患者外周血hTERT mRNA转录水平及临床意义

目的研究胃癌患者外周血中hTERT mRNA的表达，探讨其作为胃癌肿瘤细胞标志物的可行性。方法应用实时荧光定量RT—PCR技术检测96例原发性胃癌、50例胃溃疡和60例健康献血员血清中hTERT mRNA表达水平，并对阳性PCR产物进行克隆、测序。结果胃癌组hTERT mRNA表达水平（12．78±0．97）copies／ml，显著高于良性胃溃疡组（0．93±0．23）copies／ml和健康对照组（0．29±0．17）copies／ml。多变量分析表明，hTERT表达水平与患者性别、年龄、肿瘤大小、肿瘤生长方式、肿瘤分化程度、淋巴管侵犯、静脉侵犯均无显著相关性（P〉0．05），而与浸润深度、淋巴结转移、远处转移、pTNM分期及CEA有显著相关性（P〈0．05）。结论qRT—PCR具有快速、敏感和特异的特点。定量分析外周血hTERT mRNA转录水平可作为胃癌的肿瘤细胞标志物之一，用于早期诊断。     

RT-PCR检测结肠癌患者外周血CEA mRNA的意义

目的　了解结肠癌患者外周血CEAmRNA的水平。方法　采用荧光定量RT PCR技术 ,检测结肠癌患者 73例 ,健康志愿者 30例外周血CEA mRNA的水平。结果　 73例结肠癌患者外周血CEAmRNA的水平为 (10 6 0 0 .0± 2 0 6 6 7.5 )copies/ml,其中38例阳性 ,阳性率为 5 2 .1%。健康志愿者未检测到CEAmRNA。CEAmRNA水平与肿瘤病理分期和分化程度相关 ;血清CEA阳性组中CEAmRNA的阳性率显著高于CEA阴性组 (P <0 .0 1) ;结论　外周血CEA mRNA可作为结肠癌辅助诊断指标 ,定量检测外周血CEA mRNA有助于结肠癌微转移的监测     

大肠恶性肿瘤外周血微转移的检测及其临床意义

目的　研究大肠癌患者外周血CK19mRNA的表达 ,并探讨其临床意义。方法　采用RT PCR方法检测 4 1例大肠癌患者和 8例胃肠道非恶性病变手术患者的外周血CK19mRNA的表达。结果　 8例胃肠道非恶性病变手术患者、8例健康志愿者的外周血CK19mRNA无阳性表达 ;4 1例大肠癌患者外周血CK19mRNA阳性表达率分别 :单次外周血采样CK19mRNA阳性表达率为 4 1 5 % ;两次采样总阳性率大肠癌患者为 6 5 9% ,外周血重复采样阳性率高于单次采样 (P <0 0 1)。DukeC、D期患者CK19mRNA阳性表达率高于DukeB期 (P <0 0 5 )。结论　大肠癌患者外周血CK 19mRNA是一种敏感、特异的检测方法 ,可以作为检测大肠癌微转移的指标并指导其治疗和预后。     

不同手术方式对肝癌患者外周血甲胎蛋白mRNA的影响

目的通过对行腹腔镜肝癌切除术患者术前及术后外周血甲胎蛋白(AFP)mRNA的测定,探讨腹腔镜肝癌切除术对肿瘤外周血微转移的影响。方法采用实时定量的FQ-PCR方法,检测21例行腹腔镜肝癌切除术患者(腹腔镜组)术前及术后外周血AFP mRNA的表达;同时以33例开腹肝癌切除术(开腹组)及10例肝脏良性病变手术(良性对照组)作为对照,对比测定其术前术后外周血AFP mRNA的表达变化。结果术前腹腔镜组及开腹组外周血AFP mRNA表达阳性率分别为71.43%(15/21)、72.73%(24/33),明显高于良性对照组(良性对照组术前、术后均无阳性表达);术后外周血AFP mRNA表达阳性率腹腔镜组及开腹组分别为85.71%(18/21)、87.88%(29/33),与术前比较,差异均无统计学意义(χ2=1.273,P=0.259;χ2=2.395,P=0.122);术后腹腔镜组及开腹组之间比较,差异无统计学意义(χ2=0.053,P=0.817)。结论腹腔镜肝癌切除术相比传统开腹手术不增加肿瘤微转移的风险。     

Migraine and genetic and acquired vasculopathies

It is remarkable that migraine is a prominent part of the phenotype of several genetic vasculopathies, including cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL), retinal vasculopathy with cerebral leukodystrophy (RVCL) and hereditary infantile hemiparessis, retinal arteriolar tortuosity and leukoencephalopahty (HIHRATL). The mechanisms by which these genetic vasculopathies give rise to migraine are still unclear. Common genetic susceptibility, increased susceptibility to cortical spreading depression (CSD) and vascular endothelial dysfunction are among the possible explanations. The relation between migraine and acquired vasculopathies such as ischaemic stroke and coronary heart disease has long been established, further supporting a role of the (cerebral) blood vessels in migraine. This review focuses on genetic and acquired vasculopathies associated with migraine. We speculate how genetic and acquired vascular mechanisms might be involved in migraine.     

Fibrinogen and fibrin structure and functions

Fibrinogen molecules are comprised of two sets of disulfide-bridged Aalpha-, Bbeta-, and gamma-chains. Each molecule contains two outer D domains connected to a central E domain by a coiled-coil segment. Fibrin is formed after thrombin cleavage of fibrinopeptide A (FPA) from fibrinogen Aalpha-chains, thus initiating fibrin polymerization. Double-stranded fibrils form through end-to-middle domain (D:E) associations, and concomitant lateral fibril associations and branching create a clot network. Fibrin assembly facilitates intermolecular antiparallel C-terminal alignment of gamma-chain pairs, which are then covalently 'cross-linked' by factor XIII ('plasma protransglutaminase') or XIIIa to form 'gamma-dimers'. In addition to its primary role of providing scaffolding for the intravascular thrombus and also accounting for important clot viscoelastic properties, fibrin(ogen) participates in other biologic functions involving unique binding sites, some of which become exposed as a consequence of fibrin formation. This review provides details about fibrinogen and fibrin structure, and correlates this information with biological functions that include: (i) suppression of plasma factor XIII-mediated cross-linking activity in blood by binding the factor XIII A2B2 complex. (ii) Non-substrate thrombin binding to fibrin, termed antithrombin I (AT-I), which down-regulates thrombin generation in clotting blood. (iii) Tissue-type plasminogen activator (tPA)-stimulated plasminogen activation by fibrin that results from formation of a ternary tPA-plasminogen-fibrin complex. Binding of inhibitors such as alpha2-antiplasmin, plasminogen activator inhibitor-2, lipoprotein(a), or histidine-rich glycoprotein, impairs plasminogen activation. (iv) Enhanced interactions with the extracellular matrix by binding of fibronectin to fibrin(ogen). (v) Molecular and cellular interactions of fibrin beta15-42. This sequence binds to heparin and mediates platelet and endothelial cell spreading, fibroblast proliferation, and capillary tube formation. Interactions between beta15-42 and vascular endothelial (VE)-cadherin, an endothelial cell receptor, also promote capillary tube formation and angiogenesis. These activities are enhanced by binding of growth factors like fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF), and cytokines like interleukin (IL)-1. (vi) Fibrinogen binding to the platelet alpha(IIb)beta3 receptor, which is important for incorporating platelets into a developing thrombus. (vii) Leukocyte binding to fibrin(ogen) via integrin alpha(M)beta2 (Mac-1), which is a high affinity receptor on stimulated monocytes and neutrophils.     

Telemedicine and teleradiology technologies and applications

Summary. Telemedicine and teleradiology hold the key for improving future health care delivery. In this paper we first review current communication and computer technologies used in telemedicine and teleradiology. Five examples in teleradiology applications are given including hospital-integrated picture archiving and communication systems, tele-neuro-imaging, telemammography, university consortium teleradiology service, and teleradiology for second opinion. Parameters important to teleradiology applications like costs, image quality, system reliability, and turn around time are considered. Data security is discussed, including patient confidentiality and image authenticity-which will be a major issue in future teleradiology applications.     

创伤高血糖与感染和MODS早期诊断和干预

本文详细介绍了创伤后血糖应激适度理论,以及高血糖与感染和多器官功能不全综合征的关系;提出涉及胰岛B细胞功能不全的MODS实验诊断新方案和极化液个体化干预新措施,可早期发现创伤MODS、降低感染率及MODS发生率和病死率。     

腹膜后纤维化与肾积水发生关系的临床研究

目的：探讨腹膜后纤维化（RPF）导致肾积水的原因及诊治经验。方法：回顾分析2004年1月—2010年12月24例腹膜后纤维化致肾积水患者的诊治资料。结果：（1）RPF患者常见首发症状为腰背痛或腹痛（69.2%）;（2）红细胞沉降率（ESR）增快和血清IgG4升高最常见。超声检查仅提示上尿路积水。RPF的静脉肾盂造影（IVP）和CT尿路成像（CTU）表现具有特征性。IVP肾盂输尿管显影不良时,CTU能较清晰的显示上尿路影像。CT扫描发现腹膜后软组织肿块9例（37.5%）,优于超声检查;（3）输尿管松解和腹腔化手术治疗22例;行肾切除术1例;行输尿管置双J管术1例。最终确诊为继发性RPF8例,其中4例为术前诊断,3例为术中腹膜后软组织肿块冷冻活检证实,1例为术后病理证实;（4）特发性RPF手术后肾积水均获长期缓解,而继发性RPF的预后取决于原发疾病及其治疗方案。结论：影像学检查是诊断RPF的重要手段,CTU优于超声检查和IVP。输尿管松解和腹腔化手术可以使特发性RPF输尿管梗阻得到长期的缓解,术中对肿块进行冷冻活检有助于鉴别特发性和继发性RPF,及时调整治疗方案。     

探讨儿童慢性顽固性咳嗽与支原体感染的关系及临床治疗观察

目的探讨儿童慢性顽固性咳嗽与肺炎支原体（MP）感染的关系及临床疗效观察。方法采用回顾性研究方法对于现将2005年3月至2008年3月在我院的55例确诊慢性顽固性咳嗽患儿,主要表现为肺炎支原体感染为临床特点进行分析,并进一步临床治疗研究。结果①临床特点：在55例确诊慢性咳嗽的患儿中,以慢性顽固性咳嗽为主要症状。58％（32／55）的病例无肺部体征;②外周血：85％（47／55）的病例外周血变化不大,WBC（4—10）×10 9／L之间,嗜酸性粒细胞增多;③特别检查：47．27％（26／55）肺炎支原体IgM（MP—IgM）抗体阳性,83．64％（46／55）PeR技术检测肺炎支原体特异性DNA;④X光报告为多种形式。结论肺炎支原体（MP）感染是引起儿童慢性顽固性咳嗽的病因之一,对儿童慢性咳嗽,特别是顽固性咳嗽的诊治中应更加重视。     

Acetaminophen and acetylcysteine dose and duration: Past,present and future

Abstract

Acetylcysteine has been utilized successfully in the treatment of acetaminophen overdose since the 1970s. Although prospective trials as to efficacy and safety of acetylcysteine were conducted, there were no randomized controlled trials. This commentary addresses the reasons for this, and the background to choice of dose of acetylcysteine utilized in the oral and IV dosing regimens. Nomograms to predict possible hepatotoxicity based upon time of ingestion of acetaminophen were developed from a relatively arbitrary definition of toxicity as an aspartate aminotransferase/alanine aminotransferase (ALT/AST) greater than 1000 IU/L. While these have proved generally useful, patients still continue to develop hepatic damage after acetaminophen overdose, particularly if they present late after ingestion. The optimum management of these patients remains unclear, and one area of uncertainty is the dose and duration of acetylcysteine in various circumstances. This article discusses the issues that need to be elucidated to better target changes in acetylcysteine dose. The potential for measurements of other markers to improve treatment selection is the subject of further research.     

Physician and Nurse Practitioner Teamwork and Job Satisfaction: Gender and Profession

Designing interprofessional primary care teams composed of physicians and nurse practitioners (NPs) is a national priority. We assessed how profession and gender affect teamwork and job satisfaction among primary care physicians and NPs by using survey data from 186 physicians and 398 NPs practicing in New York State. Our regression models show profession (NP vs physician) moderates the associations of gender with teamwork and job satisfaction. Among NPs, men had higher job satisfaction than women. Among physicians, women had higher job satisfaction than men. Our results can benefit interprofessional primary care teams to optimize their professional and gender mix.