肺癌患者外周血微转移检测的实验研究

目的探讨肺癌患者外周血CEAmRNA、CK19mRNA和CK20mRNA的表达对诊断和监测肺癌微转移的临床意义。方法采用荧光定量聚合酶链反应(PCR)技术,检测60例不同类型和临床分期肺癌患者外周血CEAmRNA、CK19mRNA和CK20mRNA的表达水平。同时对12例肺良性病变患者和60名健康体检者外周血CEAmRNA、CK19mRNA和CK20mRNA表达水平进行检测,并分析CEAmRNA、CK19mRNA和CK20mRNA的表达对肺癌微转移诊断敏感性、准确性。结果60例肺癌患者外周血CEAmRNA的水平为(15 675±26 453)copies/ml,阳性率为(32/60)53.3%;CK19mRNA的水平为(15 213±26 326)copies/ml,阳性率为(35/60)58.3%;CK20mRNA的水平为(10 879±20 986)copies/ml,阳性率为(25/60)41.6%。42例肺良性病变中CK20mRNA阳性1例(2.3%),CEAmRNA和CK19mRNA均为阴性,60例健康体检者外周血CEAmRNA、CK19mRNA和CK20mRNA均未见表达。CEAmRNA、CK19...     

肺特异性X蛋白细胞角蛋白19mRNA在NSCLC外周血中的表达及其对微转移的诊断价值

目的检测非小细胞肺癌(NSCLC)患者外周血中肺特异性X蛋白(Lunx)mRNA、细胞角蛋白19(CK19)mRNA的表达对NSCLC微转移的诊断价值及临床意义。方法采用荧光定量反转录-聚合酶链反应(RT-PCR)技术分别对50例NSCLC患者、15例肺良性疾病(BLD)患者和10名健康人外周血中Lunx mRNA和CK19mRNA的表达进行检测分析。结果 Lunx mRNA、CK19mRNA在NSCLC外周血中的阳性表达率分别70%、68%,与其他2组比较差异均有统计学意义(P<0.05)。2种标志物共同阳性表达者有28例,阳性率为56%,2种标志物检测比较,差异无统计学意义。Lunx mRNA、CK19mRNA在NSCLC患者阳性表达率与年龄、肿瘤大小、肿瘤部位、病理类型均无关(P>0.05),而与不同临床分期、组织分化程度及有无淋巴结转移有关(P<0.05)。结论 Lunx mRNA、CK19mRNA的高表达可能与NSCLC的微转移有关,可作为检测NSCLC微转移的良好指标,联合检测有助于提高检测的准确度。     

荧光定量PCR技术检测乳腺癌患者外周血CEA、CK19mRNA表达的意义

目的探讨荧光定量PCR(FQ-PCR)检测外周血癌胚抗原(CEA，eareinoembryonic antigen)、细胞角蛋白19(CK19，cytokeratin 19)m1RNA的表达在乳腺癌外周血微转移诊断中的应用。方法采用FQPCR法，并以β-actin为内对照，测定19例乳腺癌组织中特异性mRNA表达情况，以及测定28例健康女性体检者、11例良性乳腺疾病患者和51例乳腺癌患者的外周血中CEA、CK19mRNA的表达。结果19例乳腺癌组织中CEAmRNA的阳性率为78．9％(15／19)，CK19mRNA的表达阳性率为100％(19／19)。本研究应用FQ-PCR检测外周血CEA、CK19mRNA的表达来检测乳腺癌外周血微转移，以上有一项为阳性即判定为转移。正常对照组和良性乳腺疾病组CEA、CK19mRNA的表达均为阴性，乳腺癌组显著高于前2组。结论FQ-PCR技术是高度灵敏、高度特异的快速定量检测CEA、CK19mRNA的方法，可有助于乳腺癌的诊断和评估预后。     

Lunx mRNA MMP-7 mRNA hTERT mRNA的表达对非小细胞肺癌微转移诊断的价值

目的应用实时荧光定量聚合酶链反应(RT-PCR)技术联合检测非小细胞肺癌(NSCLC)患者外周血中肺特异性X蛋白基因(Lunx)mRNA、基质金属蛋白酶(MMP)-7 mRNA、端粒酶逆转录酶(hTERT)mRNA的表达情况,探讨其作为分子标记物对于微转移的诊断价值。方法应用RT-PCR技术,检测151例NSCLC患者、24例肺良性病变和30名健康志愿者外周血Lunx mRNA、MMP-7 mRNA、hTERT mRNA水平。结果 NSCLC患者外周血Lunx mRNA、MMP-7 mRNA和hTERT mRNA的表达阳性率分别为41.7%(63/151)、34.4%(52/151)和54.3%(82/151),均明显高于肺良性患者和健康人(P<0.01)。Lunx mRNA和hTERT mRNA的表达与临床分期和淋巴结转移相关。MMP-7 mRNA的表达与临床分期相关。联合3项检测灵敏度达74.2%,准确性达77.3%。结论Lunx mRNA、MMP-7 mRNA和hTERT mRNA可作为检测NSCLC患者外周血微转移的分子标记物。     

喉癌、喉咽癌患者CK19和CK20的表达及其临床意义

目的 探讨喉癌、喉咽癌患者外周血中细胞角蛋白(CK)19 mRNA、CK 20 mRNA的表达及临床意义.方法 采用实时荧光定量RT-PCR检测50例喉癌、喉咽癌患者(A组)和20例正常人群(C组)外周血CK19和CK20 mRNA的表达水平.结果 A组外周血中CK19和CK20 mRNA表达明显高于C组(0.4626±0.1220 vs.0.1712±0.0561和0.4817±0.1401 vs.0.2175±0.0704)(P＜0.01).A组晚期患者CK19和CK20 mRNA的表达高于早期患者(P＜0.05);淋巴结转移患者外周血中CK19和CK20 mRNA表达略高于无淋巴结转移患者(P＞0.05).结论 检测外周血CK19和CK20 mRNA表达有助于喉癌、喉咽癌的诊断.     

肺腺癌组织中VEGF mRNA的表达与外周血微转移的关系

目的 探讨肺腺癌组织中血管内皮生长因子(VEGF)mRNA的表达与外周血微转移的关系.方法 应用RT-PCR方法榆测33例肺腺癌组织中VEGF mRNA和外周静脉血中CK19mRNA表达变化.另选10例正常人外周血为对照.结果 33例肺腺癌组织中VEGF mRNA的阳性率为78.79%(26/33),外周血CK19 mRNA的阳性率为57.58%(19/33);10例正常人外周血无CK19 mRNA阳性表达.肺腺癌组织中VEGF mRNA的表达与淋巴结转移和病理分期密切相关;VEGFmRNA高表达者与VEGF mRNA低表达者CK19的阳性率有显著性差异(P＜0.05).结论 肺腺癌患者组织中VEGF mRNA表达和淋巴结及血行转移关系密切;联合检测肺腺癌组织中VEGF mRNA和外周血CK19 mRNA的表达有助于评估肺癌转移、病理分期及预后.     

肺癌患者外周血Survivin mRNA和CK19 mRNA的表达及临床研究

目的通过对非小细胞肺癌（Non-small cell lung cancer,NSCLC）患者外周血生存素（Survivin）mRNA及细胞角蛋白19（CK19）mRNA的检测;探讨其对NSCLC诊断的临床价值和意义。方法采用逆转录聚合酶链反应法（RT-PCR）检测55例非小细胞肺癌患者、60例对照组的外周血Survivin mRNA和CK19 mRNA的表达情况。结果 Survivin mRNA和CK19 mRNA在NSCLC组的阳性率显著高于对照组（P〈0.01）。阳性表达率与临床TNM分期有关（P〈0.01）,与性别、年龄、病理组织类型无关（P〉0.05）。Survivin mRNA和CK19 mRNA联合检测,高于任一单一指标的检测（P〈0.05）。结论外周血Survivin、CK19 mRNA可作为早期辅助诊断NSCLC及微转移的一个有价值的肿瘤标志物。联合检测有较高的检出率。     

食管癌患者外周血CEA—mRNA、CK20-mRNA的表达及意义

目的以CEA-mRNA、CK20-mRNA为靶基因,应用逆转录聚合酶链反应（RT—PCR）的方法,检测食管癌患者肿瘤组织以及食管癌患者外周血微转移,探寻靶基因作为食管癌微转移检测的分子标志物的可行性。方法应用RT-PCR技术检测53例食管癌组织中CEA—mRNA、CK20-mRNA的表达;检测53例食管癌患者外周血中靶基因的表达,并以10例良性食管疾病患者和20名健康人外周血作为对照。结果53例食管癌患者病理组织中CEA—mRNA、CK20-mRNA的表达率为96．23％（51／53）、100％（53／53）,外周血中CEA．mRNA、CK20-mRNA表达阳性率为52．83％（28／53）、49．06％（26／53）;对照组中,10例食管良性病变患者和20名健康人外周血中均有1例CEA-mRNA表达、无CK20-mRNA表达。结论CEA—mRNA、CK20-mRNA是检测食管癌组织及食管癌外周血微转移良好的分子标志物,两者表达与食管癌TNM分期关系密切。     

大肠癌患者外周血癌细胞检测的临床意义

目的 探讨大肠癌患者外周血癌细胞的检测及临床意义。方法 以CK20mRNA为靶基因，运用逆转录一聚合酶链反应方法(RT—PCR)检测20例正常人和42例大肠癌患者术前、术后外周血中CK20mRNA表达。结果 20例正常人外周血中均无CK20mRNA表达。20例大肠癌组织标本中均有CK20mRNA表达。CK20mRNA阳性检出率与Dukes分期、淋巴结转移、肝转移存在差异有显著性。结论 RT—PCR方法检测大肠癌外周血癌细胞具有高度特异性和敏感性。外周血中CK20mRNA检测可帮助综合判断疾病恶性程度和预后，以及术后早期化疗对控制大肠癌微转移具有一定作用。     

CK19mRNA在非小细胞肺癌患者外周血中的表达及相关研究

目的：探讨细胞角蛋白19（CK19）mRNA在非小细胞肺癌患者外周血微转移的表达特点及其临床意义。方法：采用实时荧光定量RT—PCR的方法对55例非小细胞肺癌患者和20例正常人外周血CK19mRNA表达进行检测。结果：55例肺癌患者外周血液中,38例CK19mRNA表达为阳性,20例正常人外周血液中CK19mRNA均为阴性（X^2=11．35,P〈0．05）。肺癌患者外周血中CK19mRNA阳性与临床分期、病理分型和肿瘤的分化程度不相关（P＞0．05）,与淋巴结转移有关（X^2=49,P〈0．05）。结论：肺癌患者外周血中CK19mRNA表达较高可能与肺癌转移有关,检测CK19mRNA对于早期判断患者转移和复发有一定的临床意义。     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Polymorphisms in genes involved in neurotransmission in relation to smoking

Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D₁, D₂, and D₄ receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D₃ and D₅ receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking.     

Tramadol concentrations in blood and in cerebrospinal fluid in a neonate

Based on blood and cerebrospinal fluid samples collected in a full-term neonate, the penetration of tramadol in the central nervous system is described. Following intravenous administration of tramadol, a lag time of about 4 h was observed until full blood–brain equilibration was achieved. This pharmacokinetic observation is in line with a recent pharmacodynamic evaluation of the central opioid effects of tramadol in adults.     

Disease control in asthmatic children seen in private practice in Switzerland

ABSTRACT

Background: Asthma is the most common chronic childhood disease in Switzerland with a prevalence of 10%. Asthma has a high economic burden accounting for high medical costs. Assessment of disease control is likely to be of help in the implementation of strategies to improve asthma. Therefore, we aimed to evaluate asthma control and therapy regimens among children in private practice.

Methods: We assessed asthma control as well as therapy regimens in 575 asthmatic children in an experience programme in Switzerland by using an abbreviated questionnaire based on the asthma control questionnaire and the child health questionnaire on Visit 1 and Visit 2.

Results: Good asthma control at Visit 1 was only present in 25.7% of asthmatic children. Occasional asthma symptoms, limitation of physical activity, nocturnal awakening and anxiety of the parent was present in 80.5%, 41.2%, 46.8% and 57% of the children, respectively. After adjustment of therapy regimens at Visit 1, mainly by adding a leukotriene receptor antagonist, asthma control was reported to be much better in 53.4% of the children at Visit 2.

Conclusions: As asthma control is inadequately achieved within a major portion of asthmatic children, it is imperative to find measures to improve asthma control and hence, to reduce the burden of disease.