基于多重PCR技术检测蜡样芽胞杆菌3种毒素基因方法的建立及评价

目的：基于多重PCR技术,建立一种快速检测蜡样芽胞杆菌nhe、cytK和entFM 3种毒素基因的方法,并评价其效能。方法：煮沸法提取蜡样芽胞杆菌DNA,以蜡样芽胞杆菌nhe、cytK和entFM特异性毒素基因作为靶基因,采用NCBI primer-BLAST 5.0软件设计特异性引物,优化反应体系。建立三重PCR检测体系,并检测其特异性、灵敏度和稳定性。结果：煮沸法提取蜡样芽胞杆菌DNA,浓度为227 mg·L^-1,纯度为1.50～1.60。采用多重PCR体系同时检测蜡样芽胞杆菌3种毒素致病基因,于退火温度56℃时,蜡样芽胞杆菌基因引物nhe499的扩增片段为499 bp,基因引物cytK191的扩增片段为191 bp,基因引物entFM363的扩增片段为363 bp。PCR体系扩增蜡样芽胞杆菌3种毒素基因后条带清晰明亮,且无非特异性条带,与金黄色葡萄球菌、大肠埃希菌和铜绿假单胞菌均无扩增,表明蜡样芽胞杆菌基因引物nhe499、cytK191和entFM363特异性强;3种引物在同一反应体系中互不干扰。灵敏度检测,多重PCR体系最低检测限可同时达到10^...     

量子共振检测幻觉症状诊断价值的初步研究

目的 评价量子共振检测幻觉症状的诊断价值及量子医学对精神症状的诊断价值.方法 将精神科医师临床检查出的幻觉与量子共振检测结果进行了比较研究.结果 量子共振检测听幻觉具有较好的敏感度(97.6%)和准确度(89.0%),差异有显著性(x2=19.42,P<0.01).视幻觉差异无显著性(x2=0.89,P>0.05);味幻觉差异无显著性(x2=0.53,P>0.05);体感幻觉差异无显著性(x2=0.22,P>0.05).结论 量子共振检测仪将为幻听的早期发现和早期诊断提供新的依据,对精神分裂症辅助诊断具有重要的价值.     

泌尿生殖道支原体感染临床耐药性分析

目的　分析解脲脲原体(UU)和人型支原体(MH)在非淋菌性尿道炎 (宫颈炎 )患者中的感染情况及耐药现状。方法　采用珠海浪峰公司生产的支原体培养鉴定药敏试剂盒进行培养及药敏试验。结果　184例支原体培养阳性标本中,UU阳性 139例(75. 5% ),MH阳性 15例(8. 2% ),UU+MH阳性 30例 (16. 3% )。对红霉素、四环素和阿奇霉素耐药率分别为 87. 5%, 83. 7%, 58. 2%;对强力霉素、美满霉素、交沙霉素的耐药率分别为 6. 0%, 7. 1%, 15. 2%。结论　UU感染是支原体感染的主要病原体;临床治疗应首选强力霉素、美满霉素、交沙霉素。     

四-(4-三甲铵苯基)卟啉测定食品中的痕量铜

以水溶性α.β.γ.δ—四—(4—三甲铵苯基)卟啉[T-(4-TAP)P]作显色剂,测定了食品中的痕量铜。对实验条件及共存离子的干扰进行了探讨。在乙醇介质及强酸条件下测定时,其选择性及灵敏度均有显著提高。摩尔吸收系数为4.5×10~5 1mol~(-1).cm~(-1)最小检验量为:0.14ng/cm~2,回收率为96.2～106%,变异系数为:0.46～3.6%。操作简便,结果满意。     

可降解与不可降解涂层雷帕霉素洗脱支架植入后高敏C反应蛋白和白细胞介素6水平变化

目的 观察可降解涂层与不可降解涂层雷帕霉素药物洗脱支架植入后循环与局部高敏C反应蛋白和白细胞介素6水平变化.方法 连续入选118例接受PCI治疗植入1枚支架的不稳定型心绞痛患者,分为不可降解涂层组(n=65)和可降解涂层组(n=53).于术前、术后即刻、48 h、7天及9月时抽取静脉血,术前、术后即刻及术后9月冠状动脉造影随访时以微导管抽取冠状动脉局部血液,测定高敏C反应蛋白和白细胞介素6水平.结果 两组基本临床资料、PCI术情况无明显差异.两组术后即刻支架局部血清白细胞介素6水平较术前显著升高;术后48 h和7天循环高敏C反应蛋白和白细胞介素6水平明显高于术前.两组间比较,PCI术前、术后即刻、48 h、7天和9月时循环及局部高敏C反应蛋白和白细胞介素6水平均无明显差异.结论 可降解涂层与不可降解涂层雷帕霉素洗脱支架对术后循环与局部炎症反应的影响可能相似.     

氯化锂增加获得性耐药人肺癌细胞系H460R对TRAIL的敏感性

目的:通过人工诱导建立肿瘤坏死因子相关凋亡诱导配体(TRAIL)获得性耐药的肺癌细胞系,氯化锂联合rhTRAIL作用细胞,以期了解氯化锂增敏TRAIL的现象及机制。方法:通过低剂量rhTRAIL诱导人肺大细胞癌细胞系H460,建立TRAIL耐药细胞株H460R并鉴定,应用MTT和流式细胞术分析氯化锂和rhTRAIL联合给药后亲本株与耐药株细胞增殖和凋亡差异,应用RT-PCR和Western blot检测死亡受体表达。结果:rhTRAIL处理亲本株H460和耐药株H460R后细胞存活率存在显著差异(P<0.01),IC50分别为59.2ng/ml和294.8ng/ml。60ng/ml rhTRAIL处理耐药株及亲本株后平均细胞凋亡比例存在显著差异(10.5%vs 19.4%,P<0.01),耐药株表现出显著的TRAIL耐受现象。但联合20mmol/L氯化锂后,MTT及流式细胞术检测耐药株细胞增殖及凋亡发现H460R对rhTRAIL的敏感性显著增加。进一步研究发现死亡受体DR4和DR5的mRNA及蛋白水平升高,这可能是药物增敏的机制之一。结论:氯化锂能够增加获得性耐药细胞系H460R对TRAIL的敏感性,死亡受体表达增加是可能的增敏机制之一。     

Revised Japanese criteria for Sjögren's syndrome (1999): availability and validity

The Japanese criteria for diagnosing Sjögren's syndrome (SS) were revised in 1999, and consist of four major areas: histopathology, oral examination, ocular examination, and serological examination. A diagnosis of SS can be made when the patient meets at least two of these four criteria. This report describes how the revised Japanese criteria were established. After the publication of the revised Japanese criteria (1999), a research study which focused on evaluating its availability and validity was carried out in 2001 using funds from Grant-in-Aids for Scientific Research supported by the Japan Society for the Promotion of Science. The availability of the revised criteria was investigated by a questionnaire study through the Japanese Medical Society for Sjögren's Syndrome, and the use of the revised criteria for diagnosing SS in these medical facilities was found to be 76%. To evaluate the validity of the revised criteria, the records of 900 patients, including SS patients and non-SS controls, from 54 clinical centers were registered and analyzed to calculate the accuracy of the criteria. The revised Japanese criteria were found to have 96.0% sensitivity, 90.5% specificity, and 94.5% accuracy for diagnosing SS.     

Head-to-head comparison of the diagnostic accuracies of BD Veritor™ System RSV and Quidel® Sofia® RSV FIA systems for respiratory syncytial virus (RSV) diagnosis

BackgroundRespiratory syncytial virus (RSV) is one of the most common causes of severe lower respiratory tract disease among infants and young children. BD Veritor™ System RSV (BD) and Quidel^® Sofia^® RSV FIA (QD) are the new generation lateral flow digital immunoassay (DIA) tests with an instrumented read for the qualitative detection of RSV viral antigens.ObjectiveTo compare the diagnostic accuracies of BD and QD for RSV detection using fresh nasopharyngeal aspirates and nasopharyngeal swab specimens collected in universal transport media during 2013–2014 respiratory season.Study designThe two DIA tests were performed simultaneously on randomly selected specimens on a weekly basis during the RSV season until 200 fresh remnant specimens were enrolled. Real-time RT-PCR assay results were used to compare and evaluate the performance of both RSV DIA assays.ResultsAmong 200 specimens tested, RSV real-time RT-PCR assay detected RSV in 104 samples, while QD detected 84 samples and BD detected 74 samples as positive. The overall sensitivity for detection of RSV in comparison to PCR was 71.15% (61.3–79.4) for BD and 80.77% (71.6–87.6) for QD system (P = 0.36). The specificity was 100% (95.2–100) for both systems. The work flow analysis revealed that the overall specimen processing time was significantly lower for BD as compared with the QD assay.ConclusionsIn comparison with the real-time PCR, the QD system showed a higher sensitivity than that of the BD system, but the difference did not reach statistical significance (P = 0.36). Both BD and QD systems were found comparable in terms of specificity.     

Diagnostic validity (sensitivity and specificity) of panoramic X-rays in osteoarthrosis of the temporomandibular joint

Objective:

To establish the diagnostic validity of panoramic X-rays (PRx) in temporomandibular osteoarthrosis (OA) using the clinical and imaging criteria (magnetic resonance imaging, MRI) of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) as gold standards.

Methods:

Eighty-four patients with full clinical records (RDC/TMD), PRx, and MRI scans were selected. Two stomatologists evaluated the PRx for OA. The MRI findings were derived from the radiologist report. Intra/inter-examiner concordance was established. The diagnostic concordance between clinical signs and MRI was determined. Both were used as gold standards to calculate the validity of PRx in OA.

Results:

The diagnostic validity of PRx with MRI as gold standard was sensitivity?=?69·0% and specificity?=?67·9%. The diagnostic validity of PRx with clinical criteria as gold standard was sensitivity?=?61·6% and specificity =?57·9%.

Discussion:

Panoramic X-rays have scant diagnostic validity in temporomandibular osteoarthrosis when taking MRI or clinical criteria of RDC/TMD as gold standards.