人胸膜间皮细胞水通道蛋白1～10 mRNA的表达

体外培养人胸膜间皮细胞(HPMC),检测人胸膜间皮细胞水通道蛋白1～10mRNA的表达,探讨其在胸腔内液体平衡中的意义.从胸腔积液中分离人胸膜间皮细胞,进行培养,用形态学和免疫组化染色进行细胞鉴定.用RT-PCR检测水通道蛋白1～10(AQP1～10)mRNA在人胸膜间皮细胞上的表达.成功建立人胸膜间皮细胞体外培养模型,鉴定证实为间皮细胞,人胸膜间皮细胞上AQP1～10mRNA均有表达,AQP1、AQP9、AQP10表达丰富.人胸膜间皮细胞存在AQP1～10mRNA的表达,结合已知水通道蛋白的功能,证实人胸膜间皮细胞参于胸腔内液体转运.     

大鼠肺泡Ⅱ型上皮细胞的原代分离

目的建立大鼠肺泡Ⅱ型上皮细胞(alveolar epithelial cells typeⅡ,AECⅡ)分离、纯化、原代培养及鉴定的方法。方法用4.2U/ml的弹性蛋白酶通过气管插管注入肺泡内,消化分离AECⅡ。把细胞悬液接种到包被有大鼠IgG的塑料平皿中纯化细胞。用电镜、碱性磷酸酶显色法、改良巴氏染色法、单宁酸染色法、免疫组化染色法鉴定AECⅡ。结果细胞纯度达到90%以上,倒置显微镜下可见细胞呈岛屿状生长。电镜下可见细胞内有大量板层小体,包膜上有绒毛结构。碱性磷酸酶染色法(BCIP/NBT)可见胞浆内有蓝色颗粒。改良巴氏染色法、单宁酸染色法可见胞浆内有黑色颗粒。抗大鼠肺泡表面活性蛋白A(surfactant protein A,SP-A)免疫组化染色呈阳性反应。结论弹性蛋白酶作用温和,不损伤胞膜,分离所得细胞活力好;IgG免疫粘附纯化法操作简单,纯化效率高。电镜、BCIP/NBT、巴氏染色、单宁酸染色及免疫组化染色等鉴定方法稳定可靠,特异性高。     

家兔肋胸膜淋巴孔的发现

为了研究成年家兔肋胸膜淋巴孔的超微结构与三维构形 ,作者应用扫描电镜和透射电镜对成年家兔肋胸膜淋巴孔进行观察 ,用计算机图像处理系统对胸膜淋巴孔作图像数据化处理 ;NaOH溶液消化间皮细胞 ,裸露间皮下结缔组织和筛斑 (maculacribriform)。发现肋胸膜立方形间皮细胞 (cuboidalmesothelialcell)之间有圆形或椭圆形的胸膜淋巴孔 (pleurallymphaticstomata) ,其平均面积和平均密度分别为 :7 2 0± 3 6 9μm2 和 1 2 1±0 72个 / 0 0 1mm2 。扁平形间皮细胞表面未见淋巴孔。胸膜淋巴孔籍胸膜下小管与毛细淋巴管相通。仅在立方形间皮下结缔组织中发现有筛斑。肋胸膜上还可见闭合淋巴孔 (closedlymphaticstomata)和由巨噬细胞组成的乳斑 (milkyspot)。覆盖在肋骨上的胸膜无淋巴孔分布。家兔胸膜淋巴孔通过胸膜下小管与淋巴管直接相连 ,形成从胸膜腔至脉管系的惟一直接通路。     

应用电视胸腔镜诊治病因不明胸腔积液的疗效观察

目的:探讨电视胸腔镜(video-assisted thoracoscopic surgery VATS)在诊治病因不明胸腔积液中的应用价值.方法:回顾分析2005年4月～2011年4月196例病因不明胸腔积液经电视胸腔镜手术的临床资料.均应用电视胸腔镜进行探查,根据病变情况选择切口部位.排净胸腔积液后,分离粘连,进行胸膜活检后恶性患者行胸膜固定术.结果:196例均明确诊断:140例恶性胸腔积液,36例结核胸腔积液,20例炎性胸腔积液.胸腔镜手术178例,胸腔镜辅助胸壁小切口手术18例.手术时间30～75min,平均54 min.出血量10～120mL,平均53 mL.10例出现术后肺漏气,胸腔引流量＜50 mL/24h拔除胸腔引流管,胸管留置时间4～19天,平均9.4天.191例成功控制胸腔积液,全组无院内死亡.22例接受化疗的恶性胸腔积液患者,随访14～34个月,平均23个月,复查胸片显示无胸腔积液、积气.结论:电视胸腔镜安全、有效、微创,便于操作,可作为诊治病因不明胸腔积液的主要方法.     

利用气液界面无血清培养原代兔气管上皮细胞的研究

用低温酶消化法分离兔气管上皮细胞,具有细胞损伤小,活力及纯度高的优点,成纤维细胞污染极低。人胎盘胶原提高了气管上皮细胞贴壁性。无血清培养基能促进细胞增殖,分化和成熟。气液界面培养方式更好地模拟了气管上皮细胞的天然生长环境,在膜上呈复层生长,有利于细胞的分化成熟及功能表达。光镜下细胞形态及免疫组化细胞角蛋白染色阳性证实培养细胞为气管上皮细胞。本文所建立的兔气管上皮细胞体外气液界面无血清培养方法为研究气 管上皮细胞的生理和病理提供了一个十分有用的模型。     

小鼠胸膜纤维化模型中CX43蛋白的动态表达

目的研究博莱霉素诱导的小鼠胸膜纤维化模型中间隙连接蛋白CX43的动态表达变化,以阐明胸膜损伤而导致纤维化的机制。方法采用胸膜腔内注射博莱霉素和碳粉复制小鼠胸膜纤维化模型,分别于注射后第2,7,21d取肺及胸膜组织,免疫组化法测定CX43蛋白表达和Masson染色检测胸膜胶原表达情况。结果第2d胸膜开始增厚,以炎症细胞浸润为主,第7d胸膜增厚达到高峰,第21d胸膜变薄;胶原表达则从第7d开始增多,21d达高峰;CX43表达第2d增多,第7d达高峰,第21d较低,但均高于对照组（P〈0．05）。结论小鼠胸膜纤维化模型中,胸膜组织CX43的表达上调,并且与胸膜炎症反应有关。     

大鼠主动脉内皮细胞的分离培养和鉴定

目的:探索大鼠主动脉原代内皮细胞体外培养方法,为体外研究提供细胞模型。方法:分离大鼠主动脉,直接贴壁于培养皿中,荧光倒置显微镜观察细胞形态,免疫组化Ⅷ因子相关抗原染色鉴定细胞。结果:约24小时组织块边缘有游离的新生细胞长出,7天即融合成片。消化传代后细胞呈短梭形或三角形,单层生长,铺路石状,Ⅷ因子表达阳性,呈指数增殖。冻存后复苏细胞活性均超过90%。结论:用贴壁法成功建立了大鼠血管内皮细胞体外培养方法,冻存细胞存活率高,为体外研究提供了稳定的模型。     

大鼠输精管平滑肌细胞的体外培养及形态学特征(英文）

建立大鼠输精管平滑肌细胞的培养方法。取大鼠输精管,剥离外膜和内膜,用组织块法进行体外培养。用抗α-SMA(anti α-smooth muscle actin)免疫组化染色的方法鉴定培养的细胞。结果显示,在倒置显微镜下观察细胞形态多样,表现为长梭形或星形,细胞伸出突起互相接触,彼此融合,部分区域细胞多层重叠,部分区域细胞单层高低起伏,呈"峰-谷"状生长。免疫组化染色鉴定呈阳性反应,用该方法所分离、培养的输精管平滑肌细胞纯度达99%以上。应用组织块法培养大鼠输精管平滑肌细胞,操作简单,结果稳定。     

人卵巢颗粒细胞的体外培养方法探讨

目的建立人卵巢颗粒细胞分离纯化、体外培养的有效方法。方法收集体外受精—胚胎移植(IVF-ET)穿卵时的卵泡液,用胰蛋白酶消化法及密度梯度离心法分离纯化颗粒细胞并用不同培养基进行培养。结果用体积分数为50%的Percoll细胞分离液分离,DMEM/F12或McCoy’5a液体培养基进行培养,细胞纯度高,存活率高,后续生长良好。结论建立了人卵巢颗粒细胞体外培养的稳定模型,为颗粒细胞的体外研究奠定良好的基础。     

人颈椎间盘髓核细胞的体外培养和鉴定

目的：建立人颈椎间盘髓核细胞体外培养体系,并对其细胞表型进行鉴定。方法：采用酶消化法分离人颈椎间盘髓核细胞,进行单层培养,倒置相差显微镜观察细胞生长和形态,流式细胞仪测定细胞周期和凋亡率,并行甲苯胺蓝、Ⅱ型胶原及CK8免疫组化染色对其细胞表型进行鉴定。结果：原代髓核细胞凋亡率6.1±1.4%,S期细胞比例7.3±0.5%。贴壁后形态为多角形或短楔形,传代后生长加速。细胞呈甲苯胺蓝异染性;Ⅱ型胶原免疫组化染色阳性;只有少量椭圆形大细胞CK8免疫组化染色阳性。结论：成功建立人颈椎间盘髓核细胞体外培养模型,并证实成年后髓核内仍有少量细胞保持脊索细胞表型。     

Assessment of the N-PCR Assay in Diagnosis of Pleural Tuberculosis: Detection of M.tuberculosis in Pleural Fluid and Sputum Collected in Tandem

Background

The nonspecific clinical presentation and paucibacillary nature of tuberculous pleuritis remains a challenge for diagnosis. Diagnosis of tuberculous pleural effusion depends on the demonstration of the presence of tubercle bacilli in the sputum, pleural fluid, or pleural biopsy specimen, or demonstration of granuloma in pleura by histological examination. We examined the clinical utility of the diagnosis of pleural tuberculosis using the in house N-PCR assay, AFB smear microscopy and culture. Besides pleural fluid the inclusion of sputum in the efficacy of diagnosis of pleural tuberculosis was scrutinized.

Methodology/Principal Findings

Pleural fluid and sputum samples of 58 tuberculous and 42 non-tuberculous pleural effusion patients were processed for AFB smear microscopy, culture and the N-PCR assay. Mycobacteria were detected exclusively in tuberculous pleural effusion samples. None of the non-tuberculous pleural effusion samples were positive for mycobacteria. Comparative analysis showed that the N-PCR assay had the highest sensitivity. Inclusion of sputum along with pleural fluid increased N-PCR sensitivity from 51.7 to 70.6% (p<0.0001).This improved sensitivity was reflected in AFB smear microscopy and isolation by culture. The sensitivity enhanced on inclusion of sputum from 3.4 (p = 0.50) to 10.3% (p = 0.038) for AFB smear microscopy and for isolation of mycobacteria from 10.3(p = 0.03) to 22.4% (p = 0.0005). Thirteen isolates were obtained from 58 pleural tuberculosis patients. Eleven mycobacterial isolates were identified as M.tuberculosis and two as M.fortuitum and M.chelonae. Complete concordance was seen between the biochemical identification of isolates and the N-PCR identification of mycobacterial species prior to isolation.

Conclusions/Significance

To the best of our knowledge this is the first PCR based report on utility of sputum for diagnosis of pleural tuberculosis. The present study demonstrates that a combination of pleural fluid with sputum sample and N-PCR improved the diagnosis of pleural tuberculosis.     

Cytology of pleural effusion associated with disseminated infection caused by varicella-zoster virus in an immunocompromised patient. A case report

BACKGROUND: Pleural effusion caused by varicella-zoster virus (VZV) is rare. We report a case of a woman with acute lymphocytic leukemia (ALL) who developed a pleural effusion caused by VZV infection. CASE: A 55-year-old woman with ALL treated with consolidation therapy developed skin vesicles and a pleural effusion. Pleural fluid smears contained numerous mesothelial cells, which had ground-glass nuclei or eosinophilic nuclear inclusions. Some multinucleated giant cells were also seen. Electron microscopic examination revealed intranuclear virus particles, about 150 nm in diameter, in some mesothelial cells. Tissue samples from the skin, lungs, pleura, liver, pancreas, kidneys and gastrointestinal tract, obtained at autopsy, contained many virus-infected cells. They were positive for VZV glyco-protein 1 by immunohistochemistry. CONCLUSION: VZV infection should be considered in the differential diagnosis of an unexplained exudative pleural effusion, especially in immunocompromised hosts.     

Inflammatory responses in experimental tuberculosis pleurisy

A model of tuberculous pleurisy in New Zealand white rabbits was developed to describe the sequential cellular and biochemical changes in pleural fluid. Bacille Calmette-Guérin (BCG) in 4 X 10(7) colony-forming units was introduced into the right pleural space of rabbits previously sensitized by intradermal BCG. Pleural fluid was obtained via serial thoracenteses. A normal-pH, normal-glucose, exudative effusion was seen through 144 hours. Polymorphonuclear leukocytes were the first cells to respond to the introduction of tubercle bacilli in the pleural space; they remained the predominant cell for the first 24 hours and were followed by macrophages, which peaked at 96 hours, and then by lymphocytes. Numerous granulomata were observed on both the visceral and parietal pleura ten days following intrapleural instillation of BCG. We propose that the polymorphonuclear leukocyte influx is not a nonspecific response to pleural injury and that such a leukocyte response, either itself or through its interaction with the macrophage, plays a role in host defense mechanisms against the tubercle bacilli.     

Ultrasonic diagnosis of pleurisy

Ultrasound investigation of changes in the pleural cavity was conducted in 146 patients with infectious and infectious-allergic pleuritides, beginning with the first clinical manifestations to the outcome of the disease. The following stages were noted: thickening of the pleural layer in a limited area, sometimes of the endothoracic fascia and connective tissue layer between it and the parietal pleura; the appearance of fibrinous effusion; fluid accumulation; encapsulation and multilocular pleurisy; exudate resorption, fibrinolysis; fibrin involvement by connective tissue and formation of adhesions. Besides, an ultrasound picture of the normal chest wall and pleural cavity is presented.     

252 例胸膜恶性肿瘤的临床病理分析

目的：探讨胸膜恶性肿瘤的病理类型、肿瘤所占比例、临床病理特征及鉴别诊断。方法：结合病理形态学及免疫组化方法对 252 例胸膜恶性肿瘤进行诊断及鉴别诊断。结果：252 例胸膜恶性肿瘤包括胸膜穿刺活检120 例,胸腔镜活检25 例,伴有胸膜转 移的恶性胸水107 例;男性143 例,女性109 例,年龄19-87 岁,平均年龄59.9 岁。临床主要症状是胸闷、气短、咳嗽、胸痛等。CT 表现为胸膜增厚、胸水(90%)、多发或单发胸膜结节和原发器官占位性病变。活检病例中,转移性癌86 例(34.1%),包括肺腺癌64 例(25.4%),小细胞癌11 例(4.4%),鳞癌11 例(4.4%),恶性间皮瘤47 例(18.7%),滑膜肉瘤9 例(3.6%),非霍奇金淋巴瘤3 例(1.2%); 恶性胸水病例病例中转移性癌95 例(37.7%),包括肺腺癌85 例(33.7%),小细胞癌6 例(2.4%),鳞癌2 例(0.8%),乳腺腺癌2 例 (0.8%),恶性间皮瘤8 例(3.2%),非霍奇金淋巴瘤4 例(1.6%)。结论：胸膜恶性肿瘤中以转移性腺癌多见,其次为恶性间皮瘤,结合 形态学及免疫组织化学检测不同标志物的表达有助于诊断胸膜恶性肿瘤的种类。     

A Simple Method for Differentiating Complicated Parapneumonic Effusion/Empyema from Parapneumonic Effusion Using the Split Pleura Sign and the Amount of Pleural Effusion on Thoracic CT

Background

Pleural separation, the “split pleura” sign, has been reported in patients with empyema. However, the diagnostic yield of the split pleura sign for complicated parapneumonic effusion (CPPE)/empyema and its utility for differentiating CPPE/empyema from parapneumonic effusion (PPE) remains unclear. This differentiation is important because CPPE/empyema patients need thoracic drainage. In this regard, the aim of this study was to develop a simple method to distinguish CPPE/empyema from PPE using computed tomography (CT) focusing on the split pleura sign, fluid attenuation values (HU: Hounsfield units), and amount of fluid collection measured on thoracic CT prior to diagnostic thoracentesis.

Methods

A total of 83 consecutive patients who underwent chest CT and were diagnosed with CPPE (n=18)/empyema (n=18) or PPE (n=47) based on the diagnostic thoracentesis were retrospectively analyzed.

Results

On univariate analysis, the split pleura sign (odds ratio (OR), 12.1; p<0.001), total amount of pleural effusion (≥30 mm) (OR, 6.13; p<0.001), HU value≥10 (OR, 5.94; p=0.001), and the presence of septum (OR, 6.43; p=0.018), atelectasis (OR, 6.83; p=0.002), or air (OR, 9.90; p=0.002) in pleural fluid were significantly higher in the CPPE/empyema group than in the PPE group. On multivariate analysis, only the split pleura sign (hazard ratio (HR), 6.70; 95% confidence interval (CI), 1.91-23.5; p=0.003) and total amount of pleural effusion (≥30 mm) on thoracic CT (HR, 7.48; 95%CI, 1.76-31.8; p=0.006) were risk factors for empyema. Sensitivity, specificity, positive predictive value, and negative predictive value of the presence of both split pleura sign and total amount of pleural effusion (≥30 mm) on thoracic CT for CPPE/empyema were 79.4%, 80.9%, 75%, and 84.4%, respectively, with an area under the curve of 0.801 on receiver operating characteristic curve analysis.

Conclusion

This study showed a high diagnostic yield of the split pleura sign and total amount of pleural fluid (≥30 mm) on thoracic CT that is useful and simple for discriminating between CPPE/empyema and PPE prior to diagnostic thoracentesis.     

Fine needle aspiration of follicular variant of papillary thyroid carcinoma presenting with pleural effusion: a case report

BACKGROUND: Malignant pleural effusion in association with mesothelioma, bronchogenic carcinoma and breast carcinoma is common, although less frequently reported with other malignancies. We report a follicular variant of papillary thyroid carcinoma (FVPTC), diagnosed on fine needle aspiration cytology (FNAC) of thyroid and lymph nodes and subsequently proved to have metastasized to the pleural cavity. CASE: A 46-year-old man presented with history of breathlessness, thyroid swelling, pleural effusion and bilateral cervical lymphadenopathy. FNAC of the thyroid swelling and the lymph nodes showed features of FVPTC with cervical lymph node metastasis. Pleural fluid examination led to suspicion of pleural involvement by metastatic deposit, confirmed by subsequent pleural biopsy. CONCLUSION: Thyroid malignancies presenting with pleural effusion are rare. In this case, although pleural fluid cytology suggested involvement of pleura, a definitive diagnosis could be rendered only on pleural biopsy. An ancillary aid, such as immunocytochemistry, could have helped establish pleural involvement on routine pleural fluid cytology alone. This case emphasizes the possible existence of rare cases of FVPTC that may be associated with a dismal prognosis. In our case, initial diagnosis of FVPTC could be made only on correlating FNA features of thyroid aspirate with those of lymph node aspirate.     

252例胸膜恶性肿瘤的临床病理分析

目的：探讨胸膜恶性肿瘤的病理类型、肿瘤所占比例、临床病理特征及鉴别诊断。方法：结合病理形态学及免疫组化方法对252例胸膜恶性肿瘤进行诊断及鉴别诊断。结果：252例胸膜恶性肿瘤包括胸膜穿刺活检120例,胸腔镜活检25例,伴有胸膜转移的恶性胸水107例;男性143例,女性109例,年龄19—87岁,平均年龄59．9岁。临床主要症状是胸闷、气短、咳嗽、胸痛等。CT表现为胸膜增厚、胸水（90％）、多发或单发胸膜结节和原发器官占位性病变。活检病例中,转移性癌86例（34．1％）,包括肺腺癌64例（25．4％）,小细胞癌11例（4．4％）,鳞癌11例（4．4％）,恶性间皮瘤47例（18．7％）,滑膜肉瘤9例（3．6％）,非霍奇金淋巴瘤3例（1．2％）;恶性胸水病例病例中转移性癌95例（37．7％）,包括肺腺癌85例（33．7％）,小细胞癌6例（2．4％）,鳞癌2例（0．8％）,乳腺腺癌2例（0．8％）,恶性间皮瘤8例（3．2％）,非霍奇金淋巴瘤4例（1．6％）。结论：胸膜恶性肿瘤中以转移性腺癌多见,其次为恶性间皮瘤,结合形态学及免疫组织化学检测不同标志物的表达有助于诊断胸膜恶性肿瘤的种类。     

Chylothorax in rhesus monkeys with intravenous catheters.

Chylothorax associated with the use of indwelling intravenous catheters was diagnosed in 13 Macaca mulatta. Clinical signs were marked respiratory embarrassment or sudden death. Lesions at necropsy included large quantities of sterile pleural fluid, pulmonary atelectasis and chronic fibrinous pleuritis. Lipid was present in the effusion and in tissue sections of visceral pleura.