结直肠癌变过程中增殖诱导配体及其受体表达水平分析

目的分析结直肠癌发生各阶段组织中增殖诱导配体(APRIL)及其受体mRNA的表达水平。方法在APRIL及其受体基因的高保守区设计相应引物和荧光探针,实时检测PCR产物的荧光强度,根据标准品建立标准曲线,由软件自动计算出待测样本中靶基因mRNA的准确含量,并以靶基因和内参β2微球蛋白(β2M)mRNA含量比值作为评价靶基因表达水平的指标。结果批内和批间重复性测定的变异系数(cv)分别为6.52%~12.02%和8.76%~14.16%。APRIL在中、重度异型增生肠组织中的表达水平显著高于正常肠粘膜(P<0.05),在原位癌、浸润癌组织中的表达水平又显著高于正常肠粘膜(P<0.05),而其受体B细胞成熟抗原(BCMA)和穿膜蛋白活化物(TACI)在各种胃粘膜病变之间表达水平差异无统计学意义(P>0.05)。结论APRIL在结直肠癌病变演进过程中呈现累积和渐进趋势,可能在结直肠癌发生、发展过程中起重要作用。     

动脉粥样硬化兔血管平滑肌大电导钙激活钾通道KCMB1基因的mRNA表达量的变化研究

目的 研究动脉粥样硬化(atheresderosis,AS)时动脉血管平滑肌细胞大电导的钙激活钾通道(Large-coeduc-tanee Ca2 -activated K channel,BKca)β1亚单位(KCMB1)基因mRNA定量表达,探讨KCMB1基因在AS发生中的作用.方法 建立兔AS模型,应用逆转录.聚合酶链反应(RT-PCR),检测AS时胸主动脉KCMB1基因的mRNA表达变化.结果 大电导的钙激活钾通道KCMB1基因在动脉粥样硬化组胸主动脉中表达明显减少.结论 大电导的钙激活钾通道β1亚单位调节血管平滑肌细胞的收缩,可能与AS的发生有一定的关系.     

荧光原位杂交技术检测急性早幼粒细胞白血病的PML/RARα融合基因重排

目的：探讨双色双融合荧光原位杂交技术（dual-color and dual-fusion fluorescence in-situ hybridization,D-FISH）在急性早幼粒细胞白血病（acute promyelocytic leukemia,APL）诊断中的应用价值。方法：同时应用常规细胞遗传学R显带（RHG）核型分析和双色双融合荧光原位杂交2种方法,对21例APL患者进行检测。结果：21例APL患者中,常规细胞遗传学检出t（15;17）染色体易位18例,D-FISH检测均为PML/RARα融合基因（＋）;2例阴性和1例因无分裂相而无法分析结果者经D-FISH检测均为（＋）。D-FISH检测总阳性率100%（21/21）,高于常规细胞遗传学检查阳性率85.71%（18/21）。结论：与常规细胞遗传学方法相比,D-FISH检测APL的PML/RARα融合基因具有简便、快速、灵敏、特异性强等优点,可以作为APL临床诊断的一种重要检测方法。     

HBV-M定量与HBV-DNA检测的相关性研究

目的:探讨HBV-M定量与HBV-DNA检测的相关性.方法:采用时间分辨荧光定量法(TRFIA)和荧光定量PCR(FQ-PCR)法对360例疑似乙肝患者进行HBV-DNA和HBV-M定量检测.结果:HBeAg( )组患者血清HBVDNA检出率为95.17%,平均拷贝数为106.53±1.45/ml,HBeAg(-)组患者血清HBV-DNA检出率为53.49%,平均拷贝数为104.52±1.60/ml,HBeAg( )组的HBV-DNA阳性率,拷贝数明显高于HBeAg(-)组(P＜0.05, P＜0.01).HBsAg(-)组患者仍有HBV-DNA阳性者.结论:单凭血清学标志物模式难以判断HBV的复制程度及传染性强弱,定量检测HBV-DNA能真实反映HBV复制情况,对乙型肝炎的诊断及防治提供客观依据.     

流式细胞术在细胞凋亡检测中的应用

流式细胞仪（flow cytometer．FCM）是一项集激光、电子物理、光电测量、计算机、细胞荧光化学以及单克隆抗体技术为一体的新型高科技仪器。它可对单细胞或其他生物颗粒膜表面以及内部的化学成分进行快速定量分析与分选。随着现代技术的发展使其在分子生物学、免疫学、遗传学、临床医学、药学等领域中的应用有了突飞猛进的发展，在临床医学和科研中发挥着越来越重要的作用。     

The Clinical Usefulness of ~(99m)Tc-Tetrofosmin Scintigraphy in the Diagnosis of Lung Neoplasmas and Mediastinal Lymphoid Node Involvement

In order to investigate the clinical significance of 99mTc-Tetrofosmin （TF） scintigraphy in the evaluation of lung cancer and mediastinal lymphoid node involvement, 33 patients with pulmo- nary neoplasmas were subjected to both 99mTc-TF scintigraphies and CT scans in one week before their operations or puncturations. All the images were judged visually and the emission images were analyzed with semi-quantitative methods in addition. The results of each group were compared. There was marked difference in target/non-target （T/N） ratio between the lung cancer group and the benign lesion group （P〈0.001）. Moreover, in the lung cancer group, T/N ratio in tomographies was signifi- cantly higher than that in planar images （P〈0.01）. The sensitivity and accuracy of semi-quantitative analysis in 99mTc-TF SPECT were significantly higher than those of CT in the diagnosis of pulmonary neoplasmas （P〈0.05 and P〈0.01 respectively）, so was the sensitivity of 99mTc-TF SPECT vs CT in the diagnosis of mediastinal lymphoid node metastasis （P〈0.05）. It was also found that epidermoid squamous cell carcinomas and adenocarcinomas had a higher T/N ratio than in small cell carcinomas （P〈0.05）, and 2 h washout rate （WR） of adenocarcinomas was higher than that of epidermoid squamous cell carcinomas （P〈0.05）. In conclusion, 99mTc-TF scintigraphy showed a favorable diag- nostic accuracy in appraising lung cancers and mediastinal lymph node metastases. Furthermore semi-quantitative technology can improve the accuracy, and is potential to offer some information about histological type of the cancer tissue. Therefore, 99mTc-TF scintigraphy will be a useful tool in the diagnosis and staging of lung cancer.     

Significance of Rosiglitazone Inhibiting TLR4 Expression in Partial Hepatic Ischemia/Reperfusion of Mice

The effect of rosiglitazone as the ligand of peroxisome proliferator-activated receptor γ (PPARγ) inhibiting the TLR4 expression in ischemic lobes in partial hepatic ischemia/reperfusion injury (IRI) in BABL/C mice and the action of rosiglitazone inhibiting the TLR4 receptor-mediated inherent immune response were investigated. The model of the mouse partial hepatic ische- mia/reperfusion injury was established. All the animals were randomly divided into 3 groups: rosiglitazone group, vehicle (dimethylsulphoxide, DMSO) group and sham operation group. The hepatic samples were collected when mice were sacrificed 0, 2, 4 and 6 h after reperfusion following 1 h ischemia to analyze the acute phase of hepatic IRI. The dynamic expression of TlR4 mRNA was de- tected quantitatively by real-time-PCR, and the levels of TNF-α, IL-10 and ALT in portal vein were determined in all groups. After restoration of blood supply, the expression of TLR4 mRNA in ischemic lobes was detected in 0, 2, 4 and 6 h after reperfusion following 1 h ischemia in rosiglitazone group and vehicle group. The most intensive expression of TLR4 mRNA was present at 4 h after reperfusion in ischemic lobes in vehicle group. As compared with vehicle group, the expression of TLR4 mRNA in ischemic lobes in rosiglitazone group was significantly decreased at 4 h after reper- fusion. The level of IL-10 in portal vein was markedly up-regulated in rosiglitazone group as compared with vehicle group. Contrarily, the levels of TNF-α and ALT in portal vein were markedly down-regulated in rosiglitazone group as compared with vehicle group at every time point in mouse partial hepatic IRI model. Rosiglitazone could alleviate the hepatic IRI by inhibiting TLR4 receptor-mediated inherent immune response.     

Value of Quantitative Tissue Velocity Imaging in the Detection of Regional Myocardial Function in Dogs with Acute Subendocardial Ischemia

This study evaluated the application of quantitative tissue velocity imaging （QTVI） in assessing regional myocardial systolic and diastolic functions in dogs with acute subendocardial ischemia. Animal models of subendocardial ischemia were established by injecting microspheres （about 300 μm in diameter） into the proximal end of left circumflex coronary artery in 11 hybrid dogs through cannulation. Before and after embolization, two-dimensional echocardiography, QTVI and real-time myocardial contrast echocardiography （RT-MCE） via intravenous infusion of self-made microbubbles,were performed, respectively. The systolic segmental wall thickening and subendocardial myocardial longitudinal velocities of risk segments before and after embolization were compared by using paired t analysis. The regional myocardial video intensity versus contrast time could be fitted to an exponential function： y=A·（1-exp-β·t）, in which the product of A and β provides a measure of myocardial blood flow. RT-MCE showed that subendocardial normalized A·β was decreased markedly from 0.99±0.19 to 0.35±0.11 （P〈0.05） in 28 left ventricular （LV） myocardial segments after embolization, including 6 basal and 9 middle segments of lateral wall （LW）, 8 middle segments of posterior wall （PW） and 5 middle segments of inferior wall （IW）. However, there was no statistically significant difference in subepicardial layer before and after embolization. Accordingly, the ratio of A·β of subendocardial myocardium to subepicardial myocardium in these segments was significantly decreased from 1.10±0.10 to 0.31±0.07 （P〈0.05）. Although the systolic wall thickening did not change 5 min after the embolization in these ischemic segments （29%±3% vs 31%±5%, P〉0.05）, the longitudinal peak systolic velocities （Vs） and early-diastolic peak velocities （Ve） recorded by QTVI were declined significantly （P〈0.05）. Moreover, the subendocardial velocity curves during isovolumic relaxation predominant     

生殖泌尿道的HPV(6/11)型和HPV(16/18)型检测及结果分析

目的:了解本地区泌尿生殖道HPV(人乳头瘤病毒)(6/11)型和HPV(16/18型)感染率。方法:采用HPV(6/11)型和HPV(16/18)型荧光定量的PCR诊断试剂盒,对临床表现疑似为尖锐湿疣(CA)患者的分泌物进行FQ-PCR检测。结果:HPV(6/11)型感染阳性率为23.20%,其中女性为26.3l%,男性为19.90%;HPV(16/18)型感染阳性率为5.03%,其中女性为7.50%,男性为2.18%;HPV感染总阳性率为12.35%。结论:HPV是生殖泌尿道感染患者的重要病原,对临床表现疑似为CA患者有必要进行HPV检查,可为临床治疗提供非常有用的依据。