间质性肺疾病支气管肺泡灌洗液的酶活性研究

目的探讨支气管肺泡灌洗液（ＢＡＬＦ）多项酶活性与间质性肺疾病（ＩＬＤ）的关系。方法检测３０例ＩＬＤｓ：包括特发性肺纤维化（ＩＰＦ）１８例和结节病（Ｓａｒｃ）１２例与９例正常对照者的ＢＡＬＦ中超氧化歧化酶（ＳＯＤ）、谷胱甘肽过氧化物酶（ＧＳＨ－ＰＸ）、血管紧张素转换酶（ＡＣＥ）和乳酸脱氢酶（ＬＤＨ）活性，并分类计数ＢＡＬＦ细胞成份。结果（１）ＩＰＦ组ＢＡＬＦ中各项酶活性均与对照组间差异有显著性（ＳＯＤ和ＧＳＨ－ＰＸ降低，ＡＣＥ和ＬＤＨ升高）（Ｐ＜０．０５）；而Ｓａｒｃ组仅见ＡＣＥ明显增高（Ｐ＜０．０５）。（２）ＢＡＬＦ－ＡＣＥ与Ｓａｒｃ组淋巴细胞百分比及ＣＤ＋４／ＣＤ＋８比值均有显著线性相关（Ｐ＜０．０５）。结论ＢＡＬＦ中ＳＯＤ、ＧＳＨ－ＰＸ、ＡＣＥ和ＬＤＨ活性测定，有助于进一步探讨ＩＬＤ发病机理和提供辅助诊断依据，ＢＡＬＦ－ＡＣＥ对判断Ｓａｒｃ活动性有重要临床意义。     

间质性肺疾病血清和支气管肺泡灌洗液中Ⅲ型前胶原含量及其临床意义

检测了２６例间质性肺疾病（ＩＬＤ）患者和１１例正常对照组的血清和支气管肺泡灌洗液（ＢＡＬＦ）中Ⅲ型前胶原（ＰＣⅢ）含量，结果表明，ＩＬＤ病人血清和ＢＡＬＦ中ＰＣ－Ⅲ水平显著高于对照组；　且疾病组ＢＡＬＦ中ＰＣⅢ水平与其中巨噬细胞总数相关；ＢＡＬＦ中ＰＣⅢ含量用白蛋白标记表示，两组均分别显著高于其血清中的含量，这提示ＢＡＬＦ中高水平ＰＣⅢ是肺局部产生所致，认为ＰＣⅢ水平的改变可做为评估肺内胶原代谢的有用指标。     

卡氏肺孢子虫肺炎肺灌洗液细胞和生化成分变化的研究

目的 研究卡氏肺孢子虫肺炎（ＰＣＲ）支气管肺泡灌洗液（ＢＡＬＦ）中细胞和生化成分的变化及与细菌性肺炎的差别。方法 采用清洁级Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ大鼠５０只,每周２次皮下注射泼尼松建立ＰＣＰ模型（１４只）,阴性对照组（６只）,细菌性肺炎组（１１只）为实验组,并设正常对照组（６只）。     

荧光偏振法检测肺表面活性物质功能

荧光偏振法检测肺表面活性物质功能谢尔凡苟俊杨宗城黎鳌我们以１，６－二苯基－１，３，５－己三烯（ＤＰＨ）为荧光探针，检测了支气管肺泡灌洗液（ＢＡＬＦ）中肺表面活性物质（ＰＳ）的荧光偏振参数，以反映ＰＳ功能，并与其表面张力特性进行相关分析。试剂：用四氢呋...     

肺纤维化大鼠肺泡巨噬细胞释放肿瘤坏死因子—α和血小板…

目的 动态观察博莱霉素（ＢＬＭ）致大鼠肺纤维化过程中肺泡巨噬细胞（ＡＭ）释放肿瘤坏死因子－α－（ＴＮＦ－α）和血小板源生长因子（ＰＤＧＦ）的变化。方法 采用ＥＬＩＳＡ法测定ＴＮＦ－α；采用生物性法测定ＰＤＧＦ。结果 （１）ＡＭ释放ＴＮＦ－α于灌注ＢＬＭ后第３天开始升高，第７天达高峰，１４天后虽有下降，但仍显著高于对照组；（２）ＡＭ释放的ＰＤＧＦ于灌注ＢＬＭ后第３天升高，第７天达高峰，第１４天后下降     

肺纤维化大鼠肺泡巨噬细胞释放肿瘤坏死因子－α和血小板源生长因子的研究

目的动态观察博莱霉素（ＢＬＭ）致大鼠肺纤维化过程中肺泡巨噬细胞（ＡＭ）释放肿瘤坏死因子－α（ＴＮＦ－α）和血小板源生长因子（ＰＤＧＦ）的变化。方法采用ＥＬＩＳＡ法测定ＴＮＦ－α；采用生物活性法测定ＰＤＧＦ。结果（１）ＡＭ释放ＴＮＦ－α于灌注ＢＬＭ后第３天开始升高，第７天达高峰，１４天后虽有下降，但仍显著高于对照组；（２）ＡＭ释放的ＰＤＧＦ于灌注ＢＬＭ后第３天升高，第７天达高峰，第１４天后下降，与对照组比较差异无显著性。结论ＡＭ可能通过释放ＴＮＦ－α和ＰＤＧＦ在肺纤维化形成中发挥作用。     

卡氏肺孢子虫虫体负荷与肺损伤的关系研究

为研究卡氏肺孢子虫（Ｐｎｅｕｍｏｃｙｓｔｉｓｃａｒｉｎｉｉ,ＰＣ）虫体负荷与肺损伤的关系。采用清洁级Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ（ＳＤ）大鼠（５０只）每周两次皮下注射醋酸可的松,连续８～１３ｗｋ,建立卡氏肺孢子虫肺炎（ＰＣＰ）模型。共计１４只大鼠ＰＣＰ诱导成功,测定支气管泡灌洗液（ＢＡＬＦ）细胞计数及分类,总蛋白（ＴＰ）、白蛋白（ＡＬＢ）、碱性磷酸酶（ＡＬＰ）、乳酸脱氢酶（ＬＤＨ）、ＩＶ型胶原（Ｍ     

中国汉族系统性红斑狼疮某些易感基因的研究

为了探索我国北方汉族人群人类白细胞抗原－ＤＲ（ＨＬＡ－ＤＲ）及肿瘤坏死因子Ｂ（ＴＮＦＢ）等位基因多态性与系统性红斑狼疮（ＳＬＥ）的易感性关系，对１０６例健康人和４５例ＳＬＥ患者的ＨＬＡ－ＤＲ和对８０例健康人及４５例ＳＬＥ患者的ＴＮＦＢ等位基因，采用多聚酶链反应－限制性酶切片段长度多态性（ＰＣＲ－ＲＦＬＰ）法进行分析。结果显示：ＳＬＥ患者中频率显著升高的等位基因有ＤＲ２［Ｐ＜０．０５，相对危险性（ＲＲ）＝１．５６］及ＤＲ３（Ｐ＜０．０１，ＲＲ＝２．６９）。而ＤＲ５和对照相比呈相反结果（Ｐ＜０．０５，ＲＲ＝０．４３）。ＳＬＥ患者ＴＮＦＢ＊２等位基因频率显著增高（Ｐ＜０．０５，ＲＲ＝１．８４）。提示ＤＲ２、ＤＲ３、ＴＮＦＢ＊２可能是易感等位基因或易感等位基因标记，而ＤＲ５是一拮抗等位基因或拮抗等位基因标记。并研究了ＨＬＡ－ＤＲ、ＴＮＦＢ等位基因与患者血浆中Ｓ蛋白和补体５ｂ－９结合物的复合物（ＳＣ５ｂ－９）的水平和多种自身抗体及狼疮肾炎、狼疮肺炎、狼疮脑病等狼疮并发症之间的相关性，发现ＨＬＡ－ＤＲ２基因与狼疮肾炎的发生存在正相关（Ｐ＜０．０５，ＲＲ＝１．３２）。     

间质性肺疾病支气管肺泡灌洗液中FN和IgG的临床研究

同步检测并对比１１例正常对照者和２５例间质性肺疾病（ＩＬＤ）病人支气管肺泡灌洗液（ＢＡＬＦ）和外周血中纤维连接素（ＦＮ）和ＩｇＧ水平并探讨其意义。结果表明：（１）正常人ＢＡＬＦ和外周血中ＦＮ水平相近（Ｐ＞０．０５）；ＩＬＤ病人ＢＡＬＦ中ＦＮ水平明显增加，并高于其血浆中ＦＮ水平（Ｐ＜０．０１），而各组血浆ＦＮ水平相近（Ｐ＞０．０５）。（２）ＩＬＤ病人ＢＡＬＦ中ＩｇＧ水平明显增加，外周血中ＩｇＧ水平也略有增加。（３）ＩＬＤ病人ＢＡＬＦ中ＦＮ、ＩｇＧ均与细胞总数呈正相关，其中ＩＰＦ组ＢＡＬＦ中ＦＮ与中性粒细胞（％）呈正相关（Ｐ＜０．０５），非ＩＰＦ组ＢＡＬＦ中ＦＮ、ＩｇＧ均与淋巴细胞（％）呈正相关（ＦＮ：Ｐ＜０．０１；ＩｇＧ：Ｐ＜０．０５）。（４）ＩＬＤ病人ＢＡＬＦ中ＦＮ和ＩｇＧ之间呈明显的正相关（Ｐ＜０．０１）。以上结果提示：ＩＬＤ病人ＢＡＬＦ中ＦＮ水平明显增加，它可作为ＩＬＤ肺泡炎活动性的一个标志，而ＢＡＬＦ中ＩｇＧ水平增高可能反映ＩＬＤ体液免疫活动性的一个侧面。     

间质性肺疾病血清和支气管肺泡灌洗液中Ⅲ型前胶原含量及其临…

检测了２６例间质性肺疾病患者和１１例正常对照组的血清和支气管肺泡灌洗液中Ⅲ型胶胶原含量，结果表明，ＩＬＤ病人血清和ＢＡＬＦ中ＰＣ－Ⅲ水平显著高于对照组；且疾病组ＢＡＬＦ中ＰＣⅢ水平与其中巨细胞总数相关；ＢＡＬＦ中ＰＣⅢ含量用白蛋白标记表示，两组均分别显著高于其血清中的含量，这提示ＢＡＬＦ中高水平ＰＣⅢ是肺局部产生所致，认为ＰＣⅢ是肺局部产生所致，认为ＰＣⅢ水平的改变可做为评估肺内有原代谢的有用指     

KL-6 and surfactant proteins A and D in serum and bronchoalveolar lavage fluid in patients with acute eosinophilic pneumonia

OBJECT: The serum levels of KL-6, surfactant protein A (SP-A), and SP-D are useful biomarkers and prognostic factors for the activity of interstitial pneumonias. The aim of this study was to determine the clinical roles of the levels of KL-6, SP-A, and SP-D in the serum and bronchoalveolar lavage fluid (BALF) of patients with acute eosinophilic pneumonia (AEP). MATERIALS AND METHODS: We researched 5 cases of AEP. The levels of KL-6, SP-A, and SP-D in the sera and BALF of those patients were measured by enzyme-linked immunosorbent assay. RESULTS: KL-6 levels in BALF did not differ between AEP patients and the healthy control group, while SP-A and SP-D levels in BALF were significantly higher in the AEP patients than in the healthy control group. In sera, AEP patients had significantly higher than normal levels of SP-A and SP-D, but not of KL-6. Only in sera there was a positive correlation between SP-A and SP-D, but no apparent correlations in BALF and also between KL-6 and the others. Furthermore, the BALF levels of SP-D, but not of SP-A or KL-6, statistically correlated with the concentration of albumin in BALF. After clinical improvement, the elevated levels of serum SP-A or SP-D in AEP patients decreased until normal levels were reached within 2 months. CONCLUSION: These results suggest that the serum or BALF levels of SP-D appear to be more sensitive than those of SP-A or KL-6 at reflecting the inflammatory response in AEP lungs.     

Bronchoalveolar lavage fluid surfactant protein-A and surfactant protein-D are inversely related to inflammation in early cystic fibrosis

The pulmonary collectins surfactant protein (SP)-A and SP-D play important roles in innate lung defense, enhancing opsonization of microbes and limiting lung inflammatory responses. To quantify relationships among collectins, bacteria, and inflammation in early cystic fibrosis (CF) airway secretions, bronchoalveolar lavage fluids (BALFs) were collected from children undergoing clinically indicated bronchoscopy. Quantitative bacteriology, differential cell counts, and ELISA for SP-A and SP-D were assessed. Significantly increased numbers of neutrophils relative to bacteria were noted in BALF from CF compared with non-CF subjects. Although SP-A levels tended to be lower in CF compared with non-CF, this was only significant in the presence of bacterial infection. Among CF patients, SP-A concentrations in BALF were inversely related to inflammation, bacterial colony-forming units per milliliter, and age. SP-D levels were significantly decreased in CF patients, and SP-D was rarely detectable in the presence of infection. Among CF patients, SP-D correlated inversely with inflammation and bacterial colony-forming units per milliliter, and there was decreased immunostaining of BALF cells for SP-D in CF. Immunohistochemistry of CF autopsy lung sections for SP-A and SP-D confirmed their paucity at sites of infection and inflammation. We conclude that relative collectin deficiency occurs early in CF airways and is inversely related to inflammation in CF airways.     

肺孢子菌肺炎大鼠肺泡巨噬细胞吞噬功能及TNF-αIL-6的变化

目的观察肺孢子菌肺炎（PCP）大鼠肺泡巨噬细胞（AM）吞噬功能及支气管肺泡灌洗液（BALF）、AM培养上清TNF-α、IL-6的变化。方法 Wistar大鼠连续肌肉注射地塞米松建立免疫抑制PCP模型,检测大鼠AM吞噬功能及BALF、AM培养上清TNF-α、IL？6水平,同时设正常对照组。结果 PCP模型组大鼠AM吞噬鸡红细胞百分率为（20.61±2.04）%,吞噬指数为0.25±0.21,均明显低于对照组的（25.45±3.1）%和0.31±0.16（P均〈0.05）。BALF中TNF？α为（16.84±0.86）pg/ml,IL？6为（1.05±0.19）pg/ml,均明显高于对照组的（12.48±0.84）pg/ml和（0.86±0.11）pg/ml（P均〈0.05）;AM培养上清中TNF？α、IL？6差异均无统计学意义（P均〉0.05）。结论使用糖皮质激素造成大鼠免疫抑制诱发PCP后,大鼠AM吞噬功能降低,BALF中TNF-α、IL-6水平升高。     

免疫受损大鼠铜绿假单胞菌肺部感染后肺表面活性物质的改变

目的了解免疫受损宿主肺部铜绿假单胞菌（PA）感染后肺表面活性物质(PS)的改变。方法建立免疫受损宿主肺部PA感染的大鼠模型,同时设置对照组,在接种PA前后不同时间行左侧支气管肺泡灌洗,测定灌洗液中的总蛋白(TP)、总磷脂(TPL)、饱和磷脂酰胆碱(DSPC)、肺表面活性物质蛋白(SP-A)及右侧肺组织湿/干比。结果感染PA后,大鼠支气管肺泡灌洗液(BALF)中TPL及DSPC较感染前并无明显改变,但DSPC/TPL、DSPC/TP下降;免疫受损大鼠SP-A和SP-A/TP在感染前与对照组差别无显著性,但感染后下降明显,分别由感染前的（4.1±1.2）μg/ml、（22.5±5.0）μg/mg下降至接种后6h的（1.8±1.1）μg/ml、（1.4±0.7）μg/mg（P<0.05）,低于同期对照组大鼠水平［分别为（4.2±1.5）μg/ml,（11.7±8.1）μg/mg,P<0.05］,免疫受损大鼠TP及湿/干比升高更明显,两组大鼠SP-A与TP、湿/干比的改变呈负相关;免疫受损大鼠BALF中SP-A/TPL、SP-A/DSPC较感染前下降,感染后6h达最低,且感染后6～9h比例明显低于同期对照组。结论免疫受损大鼠PA肺部感染时,磷脂改变表现为DSPC/TPL、DSPC/TP降低,SP-A的下降与肺损伤严重程度有关,肺表面活性物质中磷脂与蛋白成分的改变不同步,SP-A下降幅度更明显。     

KL-6, surfactant protein A and D in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis.

BACKGROUND: KL-6, and surfactant protein A (SP-A) and surfactant protein D (SP-D) derived from alveolar type II cells and/or bronchiolar epithelial cells have been reported to be useful markers for interstitial lung diseases. OBJECTIVE: The aim of this study was to measure the levels of these molecules in bronchoalveolar lavage fluid (BALF) from patients with pulmonary sarcoidosis to investigate their relationship with other markers of inflammatory activity. METHODS: We measured KL-6, SP-A and SP-D levels in BALF from patients with pulmonary sarcoidosis using an ELISA. RESULTS: KL-6 and SP-D, but not SP-A levels were significantly increased in pulmonary sarcoidosis compared with controls. KL-6, SP-A and SP-D levels were significantly correlated with each other. KL-6 and SP-D levels were relatively and significantly correlated with the percentage of lymphocytes in BALF. KL-6, SP-D, but not SP-A levels were significantly correlated with the concentration of albumin in BALF. There was no significant correlation between KL-6, SP-A, or SP-D levels and chest X-ray findings, angiotensin-converting enzyme levels, or CD4/CD8 ratio in BALF. CONCLUSIONS: We conclude that KL-6 and SP-D levels in BALF were increased in pulmonary sarcoidosis. Since these markers are specifically derived from epithelial cells, it is considered that KL-6 and SP-D levels are reflecting damage or release of these markers from epithelial cells due to the inflammatory response.     

Comparative study of KL-6, surfactant protein-A, surfactant protein-D, and monocyte chemoattractant protein-1 as serum markers for interstitial lung diseases

KL-6, surfactant protein (SP)-A, SP-D, and monocyte chemoattractant protein-1 (MCP-1) are reported to be sensitive markers for interstitial lung diseases (ILD). However, each marker has been studied independently. The aim of this study was a comparative analysis of the diagnostic values of these markers. Subjects consisted of 33 patients with ILD (21 cases of idiopathic pulmonary fibrosis and 12 associated with collagen vascular diseases) and 82 control subjects (12 cases of bacterial pneumonia and 70 healthy volunteers). Receiver operating characteristic curves revealed that KL-6 was superior to the other markers. The cut-off levels for these markers that resulted in the highest diagnostic accuracy were determined to be 465 U/ml for KL-6, 48.2 ng/ml for SP-A, 116 ng/ml for SP-D, and 1080 pg/ml for MCP-1. The sensitivity, specificity, and diagnostic accuracy were 93.9%, 96.3%, and 95.7% for KL-6; 81.8%, 86.6%, and 85.2% for SP-A; 69.7%, 95.1%, and 87.8% for SP-D; and 51.5%, 92.7%, and 80.9% for MCP-1; respectively. The serum levels of SP-A and SP-D, but not of KL-6, were significantly higher in patients with bacterial pneumonia than in healthy volunteers. These results suggest that of the markers studied, KL-6 is the best serum marker for ILD.     

免疫抑制不同时间血液及BALF中细胞成分的变化与卡氏肺孢子虫及炎性细胞反应之观察

目的实验观察GC免疫抑制对卡氏肺孢子虫肺炎的发病影响。方法采用清洁级Wistar大鼠31只,26只大鼠每w 2次皮下注射地塞米松建立PCP模型,阴性对照组(5只),在第2、4、6、8w分别杀死大鼠,采集血液及BALF进行细胞计数并涂片分析血液和BALF中细胞成分。结果PCP大鼠血液中白细胞总数为(35.8±4.35)×109/L,淋巴细胞比例为(23.4±8.2)%,巨噬细胞数为(8.4±1.09)%,中性粒细胞比例为(68.2±8.35)%。PCP大鼠BALF中白细胞细胞总数为(35.24±8.10)×106/L,巨噬细胞比例为(84.9±2.49)%,淋巴细胞比例为(10.0±2.56)%,中性粒细胞比例为(5.1±1.73)%。结论PCP组中淋巴细胞减少是导致大鼠发生卡氏肺孢子虫感染的最关键的原因。大鼠BALF中巨噬细胞减少和功能减弱对卡氏肺孢子虫感染发生起主要帮助作用。中性粒细胞增加,推测和肺损伤有关。     

Surfactant proteins A and D in children with pulmonary disease due to gastroesophageal reflux

Children with gastroesophageal reflux often suffer from chronic, severe lung damage and recurrent infections. The mechanisms may involve reflux induced lung injury with alterations of the surfactant proteins (SP) SP-A and SP-D, which bind specifically to various microbes and increase their elimination by granular leukocytes and macrophages. In 20 children with gastroesophageal reflux disease (GERD) the bronchoalveolar lavage content and macromolecular organization of SP-A and SP-D was determined by enzyme linked immunosorbent assay and gel chromatography. For comparison, lavages from 17 children without respiratory diseases were investigated. Both, SP-A and SP-D were significantly reduced in children with GERD-median (25, 75 percentiles) SP-A: 362 (169, 494) ng/ml versus 867 (656, 1,761) in control subjects and SP-D: 174 (73, 456) ng/ml versus 518 (295, 748) ng/ml in control subjects. The more active, higher molecular weight oligomers of SP-A and especially those of SP-D were diminished, whereas the smaller sized forms of SP-D were markedly increased. In children with GERD, significantly reduced amounts of SP-A and SP-D and an altered structural organization of the surfactant protein oligomers were demonstrated. Such impairments of central components of the innate host defense system may contribute to the pathogenesis of the chronic lung disease commonly observed in these children.     

A case of idiopathic pulmonary upper lobe fibrosis complicated by invasive pulmonary aspergillosis]

A 72-year-old man with idiopathic pulmonary upper lobe fibrosis who had been followed for a year developed a high fever and yellow sputum in July 2001. Chest radiography and chest computed tomography (CT) showed a rapidly enlarging cavity with an internal mass and infiltration in the left upper lung field. Pulmonary aspergillosis was diagnosed by examination of bronchoalveolar lavage fluid (BALF). Administration of itraconazole improved his condition. The concentrations of surfactant proteins A (SP-A) and D (SP-D) in serum and in BALF were decreased during the clinical course. It is known that SP-A and SP-D are critical factors for host defense against aspergillus. The lowering of SP-A and SP-D in the serum and BALF seemed to reflect destructive changes of lung structure and impaired innate lung immunity that could to lead invasive pulmonary aspergillosis.