慢性肺疾病早产鼠BALF及肺组织中脂质过氧化的同步研究

目的为探讨高氧致早产儿慢性肺疾病(CLD)发生中支气管肺泡灌洗液(BALF)及肺组织中脂质过氧化的变化规律及相互间关系.方法采用高浓度氧致早产鼠CLD模型为研究对象,应用分光光度计比色法同步测定BALF和肺组织超氧化物岐化酶(SOD)活性及脂质过氧化产物丙二醛(MDA)的含量.结果两组的BALF及肺组织中SOD活性均无差异(P>0.05);而实验组,BALF及肺组织中MDA的水平呈同相变化,即吸高氧3d明显升高(P<0.05),7d达高峰(P<0.01),持续1w后逐渐下降,21d仍高于正常水平(P<0.05).结论肺组织的氧自由基损伤贯穿高氧致CLD发生、发展全过程中;BALF中MDA水平可间接反应肺组织的氧自由基损伤程度.     

海洛因对仔鼠和母鼠肺中SOD、CAT活力和MDA含量及组织结构的影响

目的 探讨海洛因对发育中仔鼠及母鼠肺组织中超氧化物歧化酶(SOD)、过氧化物氢(CAT)活力和丙二醛(MDA)含量及组织结构的影响.方法 对48例受孕小鼠(第8天开始)每天早晚分别注射0.2ml浓度为1.0g/L、1.5g/L和2.0g/L的海洛因溶液和等量的生理盐水,直到小鼠分娩,采用比色法检测胚胎15d、生后1d、7d、15d的仔鼠和母鼠肺中SOD、CAT活力及MDA含量的变化,应用生物显微技术观察各发育期仔鼠和母鼠肺组织结构的变化. 结果 海洛因对各发育期仔鼠和母鼠肺中SOD、CAT活力有显著的抑制作用(P<0.05),且抑制作用随海洛因浓度的增大而加大,但MDA含量则显著增加(P<0.05),海洛因浓度越大仔鼠和母鼠肺中MDA的含量就越高;1.5g/L和2.0g/L海洛因组各发育期仔鼠和母鼠肺泡直径和肺泡隔厚度与对照组相比显著增加(P<0.05).结论海洛因对各发育期仔鼠和母鼠肺有不同程度的损伤作用,这可能与海洛因引起肺组织中SOD、CAT活力下降和MDA含量升高有关.     

高氧致早产鼠BPD模型肺泡Ⅱ型上皮细胞形态与功能的变化

目的探讨高浓度氧致早产鼠支气管肺发育不良(BPD)形态与功能的动态变化规律.方法采用光镜、透射电镜及RT-PCR技术考察高氧致早产鼠BPD模型肺泡Ⅱ型上皮细胞形态学及SPA、SPB、 AQP5 mRNA表达动态变化.结果 1.吸氧7d出现肺泡化障碍,至21d正常肺泡结构基本消失;2.吸氧 1d,AECⅡ线粒体肿胀;3d板层小体排空;7d微绒毛减少,细胞核异染质增加.14d上述变化明显加重,线粒体嵴断裂,融解,伴有气血屏障明显增厚;至21d时细胞核固缩、融解,线粒体、板层小体、微绒毛消失,肺间质有大量纤维组织增生.3.SPA、SPB mRNA水平吸氧初期降低,随着吸氧时间的延长逐渐升高;4.实验组AQP5mRNA水平明显低于对照组.结论吸入高氧可引起肺泡Ⅱ型上皮细胞(AECⅡ)损伤,继而导致肺泡化障碍及AECⅡ分泌及转化功能变化.     

高氧对新生鼠肺组织VEGF蛋白表达及肺血管内皮细胞超微结构的影响

目的探讨高浓度氧对新生鼠肺组织血管内皮生长因子(VEGF)蛋白表达及肺血管内皮细胞超微结构影响的动态变化规律。方法建立高浓度氧诱导新生鼠CLD模型,60只新生鼠随机分为实验组和对照组,分别采用免疫组化和透射电镜技术,测定实验组和对照组在生后1d、3d、7d、14d和21dl肺组织内VEGF蛋白表达,同时观察肺血管内皮细胞超微结构变化。结果对照组肺组织VEGF蛋白表达1d主要以传导气道上皮为主,3d以后远端气道上皮表达增加,7d以后肺泡上皮和肺泡间隔明显增多,14d达高峰,以后持续高表达,实验组肺组织VEGF蛋白表达水平7d开始下降,14d以后未见阳性表达。高氧可引起肺血管内皮细胞肿胀、线粒体肿胀和毛细血管基底膜厚薄不均等各种损伤性形态变化,损伤程度随高氧时间延长而加重。结论肺血管的生长是正常肺泡发育重要环节,推测肺组织VEGF蛋白表达下降和肺血管内皮细胞的损伤在高氧诱导CLD肺血管发育障碍中可能发挥重要作用。     

慢性阻塞性肺疾病患者血液流变学指标及ET、SOD水平变化的临床观察

目的:观察慢性阻塞性肺疾病(COPD)患者血液流变学指标及血浆内皮素(ET)和血清超氧化物歧化酶(SOD)水平的变化。方法:采用自动血流变测试仪测定COPD患者血液流变学指标,用RIA法测定血浆ET和血清SOD水平,并与健康对照组比较。结果:COPD患者血液流变学指标有明显变化,低切变率和高切变率全血粘度以及血浆粘度均增高,与对照组比较差异显著(P<0.05),血浆ET浓度明显高于对照组(P<0.01),而血清SOD含量明显低于对照组(P<0.05)。结论:COPD患者血液流变学指标异常、微循环障碍以及ET和SOD血液水平的变化是其病理改变的重要环节。     

脑缺血再灌流过程中沙土鼠脑组织和血浆SOD、MDA含量的变

复制沙土鼠实验性脑缺血再灌流模型,测定了在缺血及再灌流不同阶段血清及脑组织中超氧化物歧化酶（ＳＯＤ）的活性和脂质过氧化反应最终产物（ＭＤＡ）的含量变化,结果发现单纯缺血组与再灌流各组脑组织ＭＤＡ的含量均高于正常对照组,且再灌流１ｈ、６ｈ组的ＭＤＡ含量明显高于单纯缺血组（Ｐ＜０．０５）,并有随再灌时间延长不断升高的趋势,但三个再灌流组之间比较无明显差异。单纯缺血组与再灌流各组比较脑组织ＳＯＤ的活性均较正常对照组降低,以再灌１ｈ、６ｈ最为明显,与单纯缺血组比较Ｐ＜０．０５。实验各组与正常对照组比较血浆ＳＯＤ的活性无显著差异。表明单纯缺血和再灌流后均发生了脂质过氧化反应,并认为自由基引起的损伤,在缺血后再灌流中比单纯缺血更为严重。     

蒺藜总皂甙对AMI家兔血清SOD和MDA变化的影响

目的和方法：采用结扎家兔冠状动脉左前降支造成急性心肌梗塞（AMI）再灌注损伤模型,观察蒺藜总皂甙（TTLS）对AMI的治疗作用。结果：和普萘洛尔的作用相同,所用不同剂量TTLS均能明显保护超氧化物岐化酶（SOD）的活性,明显减少丙二醛(MDA)的含量,二者呈高度负相关。结论：TTLS对家兔急性心肌梗塞的治疗作用机理与其保护SOD的活性和减少MDA的含量有一定关系。     

BALF中ET、CGRP变化与原发性支气管肺癌的相关性研究（摘要）

本文报道经纤维支气管镜(FOB)活检、刷片、灌洗等,并获得病理组织学及细胞学诊断的12例肺癌患者,并同步检测支气管肺泡灌洗液(BALF)ET、CGRP,结果:男性10例、女性2例,年龄30～63岁,平均51.7±11.7,鳞癌6例,小细胞未分化癌4例,腺癌2例,ET高表达6例占50％,CGRP升高7例占58.0％,以CGRP升高显著,P<0.001,且CGP的高表达与小细胞未分化癌有明显相关性,ET和CGRP的高表达与临床分期呈正相关、并随癌细胞扩散和转移逐渐增加,以CGRP明显。     

PBL患者检测BALF及血清中KL-6的临床意义

目的 探讨检测饲鸽者肺（PBL）患者肺泡灌洗液（BALF）及血清涎液化糖链抗原-6（KL-6）水平的临床意义。方法 选取2018年7月~2019年12月我院收治的PBL患者45例纳入病例组,选取同期收治的肺炎患者45例纳入肺炎组,另选同期收治的支气管炎患者45例纳入对照组。采用酶联免疫吸附法测定三组BALF及血清中KL-6水平,分析KL-6与PBL者肺功能指标的相关性,比较三组1秒用力呼气的容积占预计值的百分比（FEV1%）、肺活量预计值的百分比（FVC%）、一氧化碳弥散量占预计值的百分比（DLco%）。结果 病例组BALF及血清中KL-6水平均高于肺炎组及对照组,FEV1%、FVC%、DLco%均低于肺炎组、对照组,差异有统计学意义（P＜0.05）;Pearson分析显示BALF及血清KL-6水平与FEV1%、FVC%、DLco%均呈负相关（P＜0.05）。结论 PBL患者BALF及血清KL-6水平越高,患者的肺功能越差,临床上应加强对该病患者BALF及血清KL-6水平的重视程度,并采取有效措施进行干预,以改善其肺功能。     

大鼠复合内毒素血症致肺微循环变化与肺泡表面活性物质功能的关系

本实验应用大鼠烫伤复合内毒素血症的模型,制备肺组织超薄切片,进行透射电镜观察,同时测定其肺泡表面活性物质的功能;结合白细胞计数、血清和肺组织内二醛(MDA)含量以及超氧化物歧化酶(SOD)活性的测定。结果显示,肺毛细血管内粒细胞增多并粘附于内皮细胞膜;内皮细胞受损。肺泡表面活性物质功能明显降低。血清和肺组织MDA含量明显增加,SOD活性则显著下降。用SOD治疗能明显改善上述病变。本实验结果提示,肺微循环障碍是烫伤复合内毒素血症致急性肺损伤时肺泡表面活性物质功能下降的病理基础,与白细胞在肺血管内陷落、激活、释放自由基有关。SOD对此有一定的保护作用。     

Anti-citrullinated heat shock protein 90 antibodies identified in bronchoalveolar lavage fluid are a marker of lung-specific immune responses

Previous work has demonstrated a correlation between serum anti-citrullinated HSP90 antibodies and rheumatoid arthritis-associated interstitial lung disease (RA-ILD). To further investigate this potential pathogenic relationship, we used ELISA-based techniques to assess anti-citrullinated HSP90 antibody profiles in bronchoalveolar lavage fluid (BALF) of patients with different stages of RA-ILD. 9/21 RA-derived BALF specimens demonstrated IgG and/or IgA antibodies targeting citrullinated HSP90 proteins/peptides, highlighting disease specific responses (with a predilection for RA-ILD) that did not occur in IPF patients (0/5) or healthy control subjects (0/5). Comparison of antibody profiles between BALF and matching serum specimens revealed various recognition patterns favoring predominant production of anti-citrullinated HSP90 antibodies within the lung microenvironment—further supporting the connection between this antibody specificity and parenchymal lung disease. Equally important, qualitative as well as quantitative differences in anti-citrullinated HSP90 profiles between BALF and serum indicate that the lung plays a direct role in shaping the immune repertoire of RA/RA-ILD.     

Assessment of local cellular immunity in lung cancer by bronchoalveolar lavage

Summary Small cell lung cancer (SCLC) is the most malignant of the pulmonary neoplasms and is associated with a poor local cellular immune response. 16 patients with non small cell lung cancer (NSCLC) and 11 patients with SCLC underwent bronchoalveolar lavage (BAL) in the lung which harbored the tumor in order to investigate the lymphocyte surface antigens utilizing the immunoperoxidase technique. Analysis of blood lymphocytes was performed in parallel. 8 patients with previous sarcoidosis in complete remission who underwent BAL and 10 normal blood donors served as controls.Among blood lymphocytes the CD3⁺, CD4⁺ and CD16⁺ cell populations were elevated significantly and the T4/T8 ratio was elevated in NSCLC patients, but only CD16⁺ were augmented in SCLC. Cell populations expressing the activation markers transferrin (TF) receptor, interleukin-2 (IL-2) receptor and the very late antigen VAL-1 were also increased in NSCLC, while SCLC was associated with antigen distributions similar to controls. No differences between the cohorts were seen in the expression of human leukocyte antigen (HLA)-DR. In BAL the population of CD3⁺ and CD4⁺ cells were reduced in SCLC and the T4/T8 ratio was diminished in contrast to controls and NSCLC patients, whereas these two latter groups did not differ from each other. The distribution pattern of CD16, TF receptor and IL-2 receptor in the study groups resembled that of cells of the blood stream, but CD16⁺ natural killer cells were additionally down regulated to control values in SCLC. No differences were seen in the distribution of VLA-1. HLA-DR⁺ cells were clearly elevated in both cancer groups.In general NSCLC was associated with a shift to higher relative numbers of immunocompetent and activated cells. This was most probably attributable to an immune response to neoplastic growth. This shift was largely lacking in SCLC. The analysis of lymphocytes from the periphery of the target organ emerged as a sensitive tool for the study of cellular immunity in lung cancer and showed many similarities to circulating blood cells. However, the analysis of natural killer cells and HLA-DR suggested a dissection of cellular immune response between blood and lung in pulmonary cancer. A depressive interaction between the tumor and the cellular host immune response may contribute to the exceptional malignancy of SCLC.Abbreviations BAL bronchoalveolar lavage - HLA-DR human leukocyte antigen-DR - IL-2 interleukin-2 - NSCLC non small cell lung cancer - SCLC small cell lung cancer - TF transferrin - VLA-1 very late antigen-1     

Effect of transportation stress on blood and bronchoalveolar lavage fluid components in calves

The effect of transportation stress on the content of bronchoalveolar lavage (BAL) fluid of 20 male Holstein–Friesian calves, 4–10 months old (mean weight 160 kg) was studied. The calves were healthy and had no previous history of respiratory tract diseases. During a period of 42 days experiment, the calves were kept indoors and were fed alfalfa hay and corn silage ad libitum. After a period of adaptation, on day 21, BAL fluid, blood samples, and nasal swabs were taken from all calves; then, the calves were divided into three groups: experimental (ten calves), which were transported and were deprived of food and water during transportation; control 1 (five calves), which were not transported and had free access to food and water during the 12 h of transportation of the experimental group; and control 2 (five calves), which were not transported but were deprived of food and water for the same time as the experimental group. On day 26, BAL fluid samples and nasal swabs were taken from control group 1. Blood samples were collected simultaneously from all groups at 0, 1, 3, 6, and 12 h of transportation. On days 27, 31, and 42, all previous samplings (BAL fluid, blood, and nasal swabs) were conducted on the experimental group and control group 2. Cytological, biochemical, and bacteriologic examination of BAL fluid and hematological and biochemical examination of blood samples revealed that the number of red blood cells, white blood cells, neutrophils, and the levels of cortisol, packed cell volume, total protein, and fibrinogen significantly increased, but lymphocytes significantly decreased in the experimental group compared with control groups 1 and 2 on the day of transportation (p < 0.05). In addition, regarding BAL fluid content, total cell count, macrophages, neutrophils, and total protein increased in the experimental group (p < 0.05). Pasteurella multocida was isolated from BAL fluid of three calves in the experimental group after transportation. Alteration in BAL fluid components in this study may be due to a depressed efficiency of mucociliary system and/or decreased amount of alveolar spatial surfactant either or both of which may predispose affected livestock to show the presence of P. multocida in bronchoalveolar fluid.     

T-cell subtypes in bronchoalveolar lavage fluid and in peripheral blood from patients with primary lung cancer

The changes in local immunology play an important role in lung cancer development. We used bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) for the analysis of cell profiles in patients with primary lung cancer. Twenty-one patients with confirmed primary lung cancer and 13 healthy volunteers were investigated. All persons were smokers. The analysis of T-cell subsets was performed with a flow cytometry method and with the following antibodies: anti CD3, CD4, CD8, CD16, CD25, CD45, CD56, and HLA-DR. We found differences in the proportion of lymphocytes between BALF and PB, and a higher proportion of T cells and a lower proportion of B and natural-killer (NK) cells in BALF. There was a significant difference in the proportion of T-cytotoxic/suppressor lymphocytes, which was elevated in the BALF of patients and decreased in patients' PB. The T-helper:T-cytotoxic/suppressor (Th:Tc/s) ratio was significantly lower in the BALF of patients. These changes were visible in patients with a small cell type. The percentage of T cells with the alpha chain of receptor to IL-2 (IL -R) was lower in the BALF of patients than in the control group. Our observations reflect local changes in lung environment in patients affected with lung cancer.     

角化生长因子在慢性肺疾病早产大鼠的表达

目的探讨高氧致CLD早产大鼠KGF表达的变化规律.方法 60只新生早产大鼠随机分为2组,每组30只:A组-对照组;B组-高氧组(吸入95%O2).分别采用Western Blotting及RT-PCR检测肺组织KGF及其mRNA表达.结果高氧组肺内KGF及其mRNA表达高于对照组.结论高氧引起新生早产大鼠肺内KGF及其mRNA的表达随时间发生变化.     

Bronchoalveolar lavage improves diagnostic accuracy in patients with diffuse lung disease

Bronchoalveolar lavage (BAL) is an established diagnostic tool in diffuse parenchyma lung disease. The objective of the present study was designed to investigate whether immunophenotyping affects BAL results and improves diagnostic accuracy. BAL from 61 patients was included in the study. The patients were also submitted to transbronchial biopsy, with a final diagnosis of granulomatous disease [tuberculosis (TB), n = 20; sarcoidosis (SARC), n = 3; and hypersensitivity pneumonitis (HP), n = 4]; idiopathic interstitial pneumonias (IIPs) [idiopathic pulmonary fibrosis (IPF), n = 9; organizing pneumonia (OP), n = 17]; and lung cancer (LC), n = 8. Immunohistochemistry and histomorphometry were used to identify and quantify type 1 and type 2 alveolar epithelial cells, macrophages, CD3+T‐cells, CD4+T‐cells, CD8+T‐cells, and CD20+B‐cells in BAL. These markers were correlated with a database and pulmonary function tests. The cellular, inflammatory, and immune components of BAL varied among the diagnostic groups and were negatively correlated with age and smoking history. An increased quantity of lymphocyte surface markers CD3 (P < 0.05) and CD20 (P = 0.01) was seen in IIPs. Patients with a pattern of OP had a higher proportion of type 2 alveolar epithelial cells; patients with SARC had a higher density of CD20+B‐cells and CD4+T‐helper cells; and patients with HP had a higher proportion of CD8+T‐cytotoxic cells. A positive association was found between the density of type I alveolar epithelial cells and forced vital capacity. The immunophenotyping affects the cellular, inflammatory, or immune constituents of BAL and improved the diagnostic accuracy in diffuse parenchymal lung disease. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

新生儿HIE血清SOD活性和MDA浓度变化及临床意义

目的：探讨缺氧缺血性脑病（ＨＩＥ）新生儿血清超氧化物歧化酶（ＳＯＤ）活性和丙二醛（ＭＤＡ）浓度的变化及临床意义。方法：采用化学比色法分别测定２０例ＨＩＥ新生儿和２０例正常新生儿血清ＳＯＤ活性和ＭＤＡ浓度。结果：新生儿ＨＩＥ急性期血清ＳＯＤ活性明显低于恢复期及对照组（Ｐ均〈０．０１）,有非常显著差异,ＨＩＥ急性期血清ＭＤＡ浓度明显高于恢复期及对照组（Ｐ均〈０．０１）,有非常显著差异。ＨＩＥ急性期血清     

Pneumocystis jirovecii colonisation in patients with interstitial lung disease

A prospective study was conducted to determine the prevalence of colonisation by Pneumocystis jirovecii in 80 consecutive patients who required bronchoscopy and bronchoalveolar lavage (BAL) following suspicion of interstitial lung disease (ILD). The mtLSU rRNA gene of P. jirovecii was identified by nested PCR in BAL samples. Patients with ILDs were divided into three groups: group A comprised those with idiopathic interstitial pneumonias; group B comprised those with sarcoidosis; and group C comprised those with other ILDs. The overall prevalence of P. jirovecii carriage was 33.8%, with colonisation rates of 37.8%, 18.8% and 37% in groups A, B and C, respectively (p not significant). There were more smokers among the carriers, but there were no other significant differences between carriers and non-carriers. The high prevalence of P. jirovecii carriers found among immunocompetent patients with ILDs in Spain suggests a possible role of P. jirovecii in the natural history of these diseases.     

Antigenic specificity and subset analysis of T cells isolated from the bronchoalveolar lavage and pleural effusion of patients with lung disease.

Cellular infiltrates of bronchoalveolar lavage (BAL) and pleural effusion from patients with tuberculosis (TB) and lung cancer were characterized for the presence of different T cell subsets by phenotypic analysis. The specificity of the T cells for mycobacterial antigens was then compared for the two disease compartments. The composition of T cell subsets within the BAL, in contrast to pleural effusion cells (PEC), revealed evidence of sequestration of CD8+ cells. BAL T cells were found to be a predominantly CD29+ DR+ memory population of activated cells. Although polyclonal populations of BAL T cells proliferated poorly to Mycobacterium tuberculosis antigens, mycobacterial antigen-reactive monoclonal T cell populations could be derived from the alveolar compartment. Two clones were shown to recognize the 65-kD heat shock protein of mycobacteria, and one of these clones recognized a conserved sequence of the molecule. Several BAL-derived clones, responding to a mycobacterial soluble extract, did not, however, recognize purified mycobacterial antigens, previously identified as highly stimulatory for PEC-derived T cells. T cell clones, derived from PEC of two TB patients, responded to the 38-kD and 71-kD, as well as the 65-kD mycobacterial antigens. Examination of the activation requirements of BAL-derived T cell clones, specific for mycobacterial antigens, revealed that exogenous IL-2 was necessary for the T cells to sustain proliferation. This was in contrast to the mycobacterial antigen-reactive T cells cloned from PEC. These results suggest that T cell populations with distinct antigen specificities and activation requirements are present in BAL and PEC.