Differential proteome profiling of pleural effusions from lung cancer and benign inflammatory disease patients

The pleural effusion proteome has been found containing information that directly reflects pathophysiological status and represents a potential diagnostic value for pulmonary diseases. However, the variability in protein composition between malignant and benign effusions is not well understood. Herein, we investigated the changes of proteins in pleural effusions from lung adenocarcinoma and benign inflammatory disease (pneumonia and tuberculosis) patients by two-dimensional difference gel electrophoresis (2D-DIGE). Twenty-eight protein spots displayed significantly different expression levels were positively identified by MALDI-TOF-MS representing 16 unique proteins. Five identified protein candidates were further validated and analyzed in effusions, sera or tissues. Among them, hemopexin, fibrinogen gamma and transthyretin (TTR) were up-regulated in cancer samples. The effusion concentration of serum amyloid P component (SAP) was significantly lower in lung cancer patients than in benign inflammatory patients, but no differences were found in sera samples. Moreover, a Jumonji C (JmjC)-domain-containing protein, JMJD5, was observed to be down-regulated in malignant effusions, lung cancer tissues and cancer cells. These results shed light on the altered pleural effusion proteins as a useful and important complement to plasma or other routine clinical tests for pulmonary disease diagnosis.     

NSE、CEA 和CYFRA21-1 测定对胸腔积液的诊断价值

目的:探讨神经元烯醇化酶(NSE)、癌胚抗原(CEA)和细胞角蛋白19(CYFRA21-1)测定对胸腔积液的诊断价值。方法:选择我院2009年4月～2011年9月收治的胸腔积液患者89例作为研究对象,按照胸腔积液性质分为两组,良性组47例,恶性组42例,对两组的NSE、CEA和CYFRA21-1测定结果进行比较分析。结果:恶性组患者血清和胸腔积液中NSE、CEA和CYFRA21-1水平均明显高于良性组,组间差异有统计学意义(P<0.05);NSE、CEA和CYFRA21-1联合检测对恶性胸腔积液的敏感性均明显高于单独NSE、CEA和CYFRA21-1检测,差异有统计学意义(P<0.05)。结论:NSE、CEA和CYFRA21-1测定对胸腔积液具有临床诊断价值,且三者联合检测能够明显提高诊断敏感性。     

Limitations of detection of malignancy in pleural effusions using ELISA-based TRAP assay: comparison with cytological examination

OBJECTIVE: Telomerase is active in almost all cancers from various organs but is not detectable in most normal cells. Thus, telomerase activity might be a universal and specific marker for diagnosing malignancy. The aim was to evaluate the potential use of the ELISA-based TRAP assay to detect malignancy in pleural effusion, and to compare it with conventional cytological examination. METHODS: Using the ELISA-based TRAP assay, telomerase activity was examined in 94 consecutive pleural effusions submitted for cytological examination. RESULTS: According to the results of cytology, the 94 samples were divided into two groups: group I, 79 non-malignant pleural effusions, including group IA, no association with a malignant tumour, a control group (n = 63), and group IB, associated with a malignant tumour (n = 16); and group II, 15 malignant pleural effusions. Telomerase activity was detected in five of 63 samples in group IA (7.9%), four of 16 samples in group IB (25%), and six of 15 samples in group II (40%). All five false-positive effusions were from patients with tuberculosis. Comparing group II with group IA, the TRAP assay showed 40% sensitivity, 92.1% specificity, 54.5% positive and 86.6% negative predictive value, and 82.1% accuracy. However, the detection rate of the TRAP assay (88.9%) was higher than that of the cytological examination (66.7%) in lung cancer-inflicted pleural effusions. CONCLUSION: The ELISA-based TRAP assay is relatively insensitive; therefore, it is unsuitable as a routine diagnostic tool for pleural effusion. False-positive telomerase activity due to lymphocytic contamination may weaken its diagnostic value for malignant effusions in a tuberculosis-endemic area.     

地塞米松联合尿激酶治疗结核性胸膜炎的临床效果分析

目的:探讨地塞米松联合尿激酶对结核性胸膜炎的临床效果。方法:选择2013年8月到2016年5月在我院进行诊治的结核性胸膜炎患者190例,根据随机信封抽签原则分为观察组与对照组各95例,两组都给予标准抗结核治疗方案,对照组在抗结核治疗的同时给予尿激酶治疗,观察组再给予地塞米松治疗,两组都治疗1个月。治疗后,比较两组的总有效率、不良反应的发生情况、胸腔积液完全引流时间、抽出胸腔积液总量、凝血酶原时间和凝血酶时间。结果:所有患者都注射耐受良好,未见严重并发症;观察组的总有效率(88.4%)明显高于对照组(72.6%);观察组胸腔积液完全引流时间和抽出胸腔积液总量分别为7.56±2.44d和2867.33±456.10 m L,对照组分别为9.44±2.89d和1989.92±444.20 m L,观察组胸腔积液完全引流时间明显短于对照组,且抽出胸腔积液总量显著高于对照组(P0.05)。治疗后,两组的凝血酶原时间和凝血酶时间都明显高于治疗前(P0.05),且观察组显著高于对照组(P0.05)。结论:地塞米松联合尿激酶治疗结核性胸膜炎能延长凝血酶时间和凝血酶原时间,缩短胸腔积液引流时间,增加抽出胸腔积液总量,安全性和临床疗效均较好。     

采用不同方法制作胸腹水细胞块的效果比较及临床价值研究

目的:比较不同方法制作胸腹水细胞块的效果以优化胸腹水细胞块制作程序,并对其临床价值进行探讨。方法:以2014年3月至2015年3月我科收集的胸腹水标本120例为研究对象,将每样标本平均分成三组,使用三种不同方法(试管包埋法、直接离心法、细胞块试剂盒法)制作胸腹水细胞块。对不同方法制作细胞块的成功率、完整性及细胞切片恶性细胞的检出率进行考察与比较。结果:细胞块试剂盒法成功率最高,为96.67%,试管包埋法次之,成功率为92.50%,二者相比无显著差异(P0.05)。直接离心法制作成功率为80.83%,远远低于试管包埋法及细胞块试剂盒法,差异有统计学意义(P0.05)。细胞块试剂盒法制作的细胞块完整性最高,完整标本所占比例为96.67%,试管包埋法次之,完整率为94.17%,二者相比无显著差异(P0.05)。直接离心法制作完整率为68.33%,远远低于试管包埋法及细胞块试剂盒法,差异有统计学意义(P0.05)。三种方法恶性细胞检出率以细胞块试剂盒最高,与其他两种方法比较差异有统计学意义(P0.05)。结论:细胞块试剂盒法制作胸腹水细胞块具有最高的成功率及完整性,并且可以显著提高恶性细胞检出率,值得广泛使用。     

Non‐invasive optical assessment of viscosity of middle ear effusions in otitis media

Eustachian tube dysfunction can cause fluid to collect within the middle ear cavity and form a middle ear effusion (MEE). MEEs can persist for weeks or months and cause hearing loss as well as speech and learning delays in young children. The ability of a physician to accurately identify and characterize the middle ear for signs of fluid and/or infection is crucial to provide the most appropriate treatment for the patient. Currently, middle ear infections are assessed with otoscopy, which provides limited and only qualitative diagnostic information. In this study, we propose a method utilizing cross‐sectional depth‐resolved optical coherence tomography to noninvasively measure the diffusion coefficient and viscosity of colloid suspensions, such as a MEE. Experimental validation of the proposed technique on simulated MEE phantoms with varying viscosity and particulate characteristics is presented, along with some preliminary results from in vivo and ex vivo samples of human MEEs.

In vivo Optical Coherence Tomography (OCT) image of a human tympanic membrane and Middle Ear Effusion (MEE) (top), with a CCD image of the tympanic membrane surface (inset). Below is the corresponding time‐lapse M‐mode OCT data acquired along the white dotted line over time, which can be analyzed to determine the Stokes–Einstein diffusion coefficient of the effusion.     

Fluorescence microscopy of mycobacteria in pleural effusions

The diagnostic advantage of fluorescence microscopy (FM) of Papanicolaou-stained cytological specimens obtained by bronchoscopy has been described previously. This study was designed to evaluate the method's diagnostic benefit in cytological preparations of pleural effusions in cases of active pulmonary tuberculosis. In contrast to bronchial material there is no advantage in cytological evaluation of pleural effusions by FM.     

KSHV-infected PEL cell lines exhibit a distinct gene expression profile

We analyzed the gene expression profiles of lymphocyte-originated tumor cell lines - primary effusion lymphoma (PEL) cell lines, T-cell leukemia (TCL) cell lines, Burkitt lymphoma (BL) cell lines - and two sets of normal peripheral blood mononuclear cells (PBMCs) - in order to determine characteristic gene expression profiles for each of the former three groups. And we found that these cell lines showed respective typical gene expression profiles and classified into clear four groups, PEL, TCL, BL, and normal PBMCs. Two B lymphocyte-originated tumor cell lines, PEL and BL cell lines, clearly exhibited distinct gene expression profiles, respectively. Even though there was only one line that was co-infected with both Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV), KSHV seemed to govern the gene expression profile of the co-infected line. These data suggested not only that established typical tumor cell lines show a distinct gene expression profile but also that this profile may be governed by certain viruses.     

Nitric oxide synthase activity and nitric oxide level in erythrocytes of guinea pigs with experimental otitis media with effusion

Free radicals have been implicated in the pathogenesis of an increasing number of disease and inflammatory states. They may cause cell and tissue damage by chemical modification of proteins, carbohydrates, nucleotides and lipids. Under physiological conditions free radicals are parts of normal regulatory circuits and are neutralized by antioxidants. Infections are one cause of increased free radicals production. The aim of our study was to assess whether increased oxidative stress is reflected by erythrocyte nitric oxide synthase activity and nitric oxide levels in guinea pigs with experimental otitis media with effusion (n = 6) and in a control group (n = 6). Erythrocyte nitric oxide synthase activity and nitric oxide levels were measured in both groups. The nitric oxide synthase activity and nitric oxide level in the experimental otitis media with effusion were significantly higher than those of the control group. There was a significant positive correlation between the nitric oxide synthase activity and nitric oxide in the experimental otitis media with effusion group. Thus, increased nitric oxide levels may play an important role in cell and tissue damage due to experimental otitis media with effusion.     

分泌性中耳炎大鼠咽鼓管鼓室β-防御素的表达

目的检测大鼠β防御素(ratβ-defensin,rBD)在分泌性中耳炎大鼠咽鼓管鼓室的表达,探讨β防御素在分泌性中耳炎发病机制中的作用。方法排除中耳感染的清洁级SD大鼠48只,随机分为4组,前3组36只行颈部切口经右侧听泡注入脂多糖(lipopolysaccharide,LPS)溶液(1mg/mL)30μL制作分泌性中耳炎动物模型,造模后分别于第1、3、7天断头取咽鼓管鼓室黏膜;对照组12只右侧听泡注入生理盐水30μL,左侧听泡作为正常组,3d后断头取咽鼓管鼓室黏膜。逆转录聚合酶链反应(RT-PCR)检测咽鼓管鼓室rBD-1mRNA和rBD-2mRNA的表达。结果正常大鼠咽鼓管鼓室存在rBD-1和rBD-2的表达,且rBD-1的表达较rBD-2强,差异有统计学意义;造模后第1、3、7天,rBD-1表达变化不明显;rBD-2则在造模后第1、3天明显增加,差异有统计学意义,第7天渐回复到正常水平。结论在大鼠,rBD-1可能参与正常咽鼓管鼓室的防御功能,造模后rBD-2的表达增加可能与病原体入侵后的清除相关。