Determination of poor prognostic immune features of tumour microenvironment in non-smoking patients with lung adenocarcinoma.

We have previously demonstrated that the prognostic significance of tumour-infiltrating CD8+ T cells significantly differs according to histological type and patient smoking habits in non-small cell lung cancer (NSCLC). This work suggested that infiltrating CD8+ T cells may not be activated sufficiently in the immunosuppressive microenvironment in non-smokers with adenocarcinoma. To understand the immunogenic microenvironment in NSCLC, we characterised immune cells comprehensively by performing an immunohistochemical evaluation using an alternative counting method and multicolour staining method (n = 234), and assessed immune-related gene expression by using genetic analytical approaches (n = 58). We found that high infiltration of activated CD8+ T cells expressing interferon gamma (IFN-γ) and granzyme was correlated with postoperative survival in patients with non-adenocarcinoma. On the contrary, CD8+ T-cell accumulation was identified as a worse prognostic factor in patients with adenocarcinoma, particularly in non-smokers. Infiltrating CD8+ T cells were significantly less activated in this microenvironment with high expression of various immunoregulation genes. Potentially immunoregulatory CD8+ FOXP3+ T cells and immunodysfunctional CD8+ GATA3+ T cells were increased in adenocarcinoma of non-smokers. CD4+ FOXP3+ regulatory T cells expressing chemokine receptor-4 (CCR4)- and chemokine ligand (CCL17)-expressing CD163+ M2-like macrophages also accumulated correlatively and significantly in adenocarcinoma of non-smokers. These characteristic immune cells may promote tumour progression possibly by creating an immunosuppressive microenvironment in non-smoking patients with lung adenocarcinoma. Our findings may be helpful for refining the current strategy of personalised immunotherapy including immune-checkpoint blockade therapy for NSCLC.

Experimental Study of Intraparenchymal Fibrinogen and Topical Thrombin to Seal Pleural Defects.

OBJECTIVE: Fibrin sealants are used to close surgical pleural defects, but may detach, causing a postoperative air-leak. We investigated a new means of applying fibrin glue for closing pleural defects. METHODS: Pleural defects (10-mm and 4-mm diameters, respectively) were created in swine and rats via thoracotomy. They were sealed by a) injection of a fibrinogen solution into the lung parenchyma after instillation of a thrombin solution onto the pleural defect (group A), b) fibrinogen and thrombin spray (group B), c) fibrinogen instillation and a thrombin-dipped polyglycolic acid sheet (group C), or d) fibrin glue-coated collagen fleece (group D). Resistance to airway pressure was compared and the sealed areas were histologically examined. RESULTS: In group A, the minimum seal-breaking airway pressure was consistently > 40 cmH2O, versus 37.2 ± 3.6 cmH2O in group B, 37.2 ± 4.0 cmH2O in group C, and 39.0 ± 1.7 cmH2O in group D, which was statistically significant. Histologically, the fibrin layer infiltrated the lung parenchyma and covered the defect in group A, but not in the other groups. CONCLUSIONS: The intraparenchymal injection of fibrinogen combined with instillation of thrombin created an effective fibrin layer associated with early pleural regeneration that reliably prevented pleural air leaks.

Intractable Pneumothorax Treated With Endobronchial Watanabe Spigots.

Pneumothorax secondary to nontuberculous mycobacterial lung disease is often intractable and associated with high mortality. An endobronchial Watanabe spigot (EWS) is a silicon plug developed for bronchoscopic bronchial occlusion. Here we report a case of intractable pneumothorax secondary to nontuberculous mycobacterial lung disease successfully treated using EWSs. Occlusion of the multiple ectatic bronchi connected to the bronchopleural fistula was essential for optimal effect of the EWSs.

Value of the Glasgow Prognostic Score as a Prognostic Factor in Resectable Non-Small Cell Lung Cancer.

BACKGROUND: Over the past decade, the Glasgow prognostic score (GPS), which is based on serum C-reactive protein and albumin levels, has been reported to be associated with the prognosis of patients with several types of inoperable and operable cancers. However, its applicability to operable non-small cell lung cancer (NSCLC) has not yet been established. METHODS: We retrospectively collected data from patients with pathological stage I or II NSCLC who underwent complete resection. A total of 1048 patients were categorized as either GPS-0 (n = 817 [78.0%]), GPS-1 (184 [17.6%]), or GPS-2 (47 [4.5%]). Survival curves were estimated using the Kaplan-Meier method, and the Cox proportional hazard model was used to analyze the relationship between prognosis and GPS status. RESULTS: The 5-year overall survival (OS) rates were 91.2%, 78.3%, and 75.8% for GPS-0, GPS-1, and GPS-2, respectively. There were significant differences in OS between GPS-0 and GPS-1 (p < 0.001) and between GPS-0 and GPS-2 (p < 0.001). Ten variables demonstrated to be associated with OS in a univariate analysis were subjected to a multivariate analysis. The results showed that male sex (p = 0.031), vascular invasion (p < 0.001), lymph node metastasis (p < 0.001), and GPS (p = 0.025) were significantly associated with OS. CONCLUSIONS: A high GPS is significantly associated with poor OS. Although the biological mechanism that underlies this association is not clear, this inflammation-based score may be a useful indicator of the prognosis in patients with resectable NSCLC.

Ectopic Adrenocorticotropic Hormone-Secreting Bronchial Carcinoid Diagnosed by Balloon-Occluded Pulmonary Arterial Sampling.

We present the case of a 50-year-old man with Cushing syndrome caused by an ectopic adrenocorticotropic hormone (ACTH)-secreting tumor. A small nodule was located in close association with the lateral segmental branch of the pulmonary artery in the left upper lobe. Blood samples were obtained from various branches of the pulmonary artery by balloon-occluded retrograde sampling for the measurement of location-specific serum ACTH levels. After confirmation that the pulmonary nodule was responsible for the increased ACTH secretion, lobectomy was performed. This report demonstrates the usefulness of balloon-occluded retrograde pulmonary arterial sampling for the preoperative diagnosis of an ACTH-producing tumor whose diagnosis is difficult to confirm.

Comprehensive Immune Profiling of Lung Adenocarcinomas Reveals Four Immunosubtypes with Plasma Cell Subtype a Negative Indicator.

Neoplastic cancer cells and cancer stroma (including infiltrating immune cells) determine the biology and prognosis of cancer. Various types of adaptive and innate immune cells are known to infiltrate the cancer stroma. However, the patterns and spatial distribution of immune cell infiltration as well as its association with tumor histology remain poorly understood. To address these issues, we comprehensively analyzed the infiltrating immune cells present in lung adenocarcinoma. The principal types of both adaptive and innate infiltrating immune cells were immunohistochemically evaluated in the predominant histologic components of 111 lung adenocarcinomas. The same analysis was also carried out on 143 samples of histologic subtypes making up more than 20% of tumors. As a result, plasma cells and B cells with interfollicular distribution were almost exclusively observed in invasive histologic subtypes, while an increased number of mast cells were observed in noninvasive histologic subtypes. Cluster analysis revealed four distinct immunosubtypes (CD8, mast cell, macrophage/dendritic cell, and plasma cell subtypes) based on the infiltrating immune cell profiles. These immunosubtypes correlated with histologic subtypes, and univariate and multivariate analyses identified the plasma cell subtype as an independent negative prognostic factor. These plasma cells may be one of the major producers of the immunosuppressive cytokine IL35 in cancer stroma.

Impact of comorbidity index on morbidity and survival in non-small cell lung cancer.

BACKGROUND: The number of surgeries in older patients with comorbidities is constantly growing. The present study examined the impact of comorbidity on postoperative complications and long-term survival in patients with completely resected non-small cell cancer. METHODS: Between 2004 and 2008, 423 patients with non-small cell lung cancer underwent complete resection. A retrospective comparison of perioperative mortality, morbidity, Charlson comorbidity index (CCI), and postoperative length of hospital stay was performed. RESULTS: The number of patients with CCI 0, 1-2, and ≥ 3 was 226, 170 and 27, respectively. The 5-year overall survival was 88% among patients with CCI 0, and 84% in those with CCI ≥ 1 (p = 0.05) in all pathological stages. The CCI 0 group had significantly better overall survival than CCI 0 group and 30 (15%) in the CCI ≥ 1 group (p = 0.024). Length of stay was shorter in the CCI 0 group (11 ± 5 days) than in the CCI ≥ 1 group (15 ± 19 days, p = 0.015). CONCLUSIONS: A high CCI correlated with higher postoperative morbidity and longer length of stay. We identified better a prognosis in patients with CCI 0 compared to those with CCI 1-2.

Prognostic significance of hypoxic PET using (18)F-FAZA and (62)Cu-ATSM in non-small-cell lung cancer.

OBJECTIVES: Tumor hypoxia is believed to have a strong correlation with the resistance to chemoradiotherapy. Noninvasive evaluation of hypoxic status in tumors using molecular imaging has the potential to characterize the tumor aggressiveness. We evaluated the clinical usefulness of newly-developed tumor hypoxic positron emission tomography (PET) tracers in localized non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Forty-seven patients with localized NSCLC received either or both hypoxic PETs using the tracers: (18)F-fluoroazomycin arabinoside ((18)F-FAZA) (n=45) and/or (62)Cu-diacetyl-bis (N4)-methylsemithiocarbazone ((62)Cu-ATSM) (n=22). All received (18)F-fluorodeoxyglucose ((18)F-FDG) PET tracer (n=47). We examined the correlation between uptake of three PET tracers and clinicopathological factors, and evaluated their impacts on survival after treatment retrospectively. RESULTS: A couple of commonly-identified unfavorable factors such as presence of vascular invasion and pleural invasion was significantly correlated with higher uptake of these hypoxic agents as well as that of (18)F-FDG. Larger tumor diameter, high neutrophil-to-lymphocyte ratio and advanced pathological stage were also associated with accumulation of hypoxic PETs ((18)F-FAZA, p<0.01; (62)Cu-ATSM, p<0.04), but not with that of (18)F-FDG. The patients with a higher accumulation had significantly poorer overall survival [(18)F-FAZA, HR (hazard ratio), 9.50, p<0.01; (62)Cu-ATSM, HR, 4.06, p<0.05] and progression free survival ((18)F-FAZA, HR, 5.28, p<0.01, (62)Cu-ATSM, HR, 2.72, p<0.05). CONCLUSION: Both (18)F-FAZA and (62)Cu-ATSM PET provide useful information regarding tumor aggressiveness and prediction of survival among NSCLC patients. We believe these hypoxic PETs could contribute to the establishment of the optimally individualized treatment of NSCLC.

Alveolar macrophage phenotype expression in airway-instilled bone marrow cells in mice.

No uniform consensus has been established regarding post-pneumonectomy lung regeneration. This study was undertaken to determine whether airway-instilled lung- or bone marrow-derived cells are able to differentiate and reconstitute as lung component cells in the course of post-pneumonectomy lung growth. Bone marrow cells or lung cells obtained from C57 black (BL)/6-GFP mice were intratracheally instilled into C57BL/6 mice treated with left pneumonectomy and cell differentiation was examined. It is unclear whether intratracheally instilled lung or bone marrow cells differentiate into non-hematopoietic cells after pneumonectomy. However, regardless of whether pneumonectomy is performed, intratracheally instilled bone marrow cells display a surface antigen profile that is similar to alveolar macrophages. Furthermore, these newly differentiated macrophages function similarly to resident macrophages in terms of TNF-α production, suggesting that bone marrow stem cells acquire the same macrophage phenotype. In conclusion, intratracheally instilled bone marrow cells adapt to the surrounding microenvironment, directly differentiating into alveolar macrophages, and remain in the alveolar space for at least 3 months.

Dry pleurisy complicating solitary pulmonary nodules caused by Mycobacterium avium: a case report.

INTRODUCTION: Mycobacterium avium complex (MAC) lung disease presenting as a solitary pulmonary nodule (MAC-SPN) is often asymptomatic, is more common in middle to old age, and mimics lung cancer or tuberculoma. We report herein a case of MAC-SPN in an immunocompetent young adult patient, presenting with persistent chest pain and a subacutely progressive nodule with high intense (18)F-fluorodeoxyglucose uptake. Histological examination of resected specimens revealed pleurisy, which is a rare finding of MAC-SPN. CASE PRESENTATION: A 36-year-old Japanese male presented with chest pain and a subacutely progressive pulmonary nodule. Positron emission tomography-computed tomography showed high intense (18)F-fluorodeoxyglucose uptake in the nodule. Owing to his continuous chest pain and subacutely progressive nodules, wedge resection was performed using video-assisted thoracoscopic surgery. Histological examination revealed an epithelioid granuloma and pleurisy, and the lung tissue culture was positive for mycobacteria identified as M. avium. CONCLUSION: This is the first report of MAC-SPN occurring with persistent chest pain, suggesting that MAC should be considered in the differential diagnosis of a solitary pulmonary nodule, even for patients who experience persistent chest pain. As in the present case, surgical resection with video-assisted thoracoscopic surgery is a reasonable approach to the diagnosis and treatment of MAC-SPN with possible malignancy, especially as MAC can be diagnosed using resected lung tissue culture with histological confirmation.

Survival predictors after resection of lung metastases of head or neck cancers.

BACKGROUND: Pulmonary metastasectomies are performed for a variety of cancers, though few reports have examined their merit for head and neck cancers. This study examined the relationship between clinical and pathological characteristics and survival after resection of lung metastases of these cancers. METHODS: Between 1986 and 2013, 34 patients presenting with pulmonary metastases of primary head or neck cancers underwent surgical resections at our institution. We retrospectively analyzed their clinical and pathological characteristics and the patients' survival after metastasectomy in search of adverse prognostic factors. RESULTS: The primary sites of cancer were: the tongue in eight patients, the pharynx and salivary glands each in seven, the larynx in five, and miscellaneous sites in seven patients. Squamous cell carcinoma (SCC) was present in 19, adenoid cystic carcinomas in 10, and other diagnoses in five patients. The median disease-free interval (DFI) between the initial surgery and the metastasectomies was 40 months. The overall five-year survival rate was 57.9%, and median survival time was 77 months. By single variable analysis, a DFI of ≤26 months, age ≥60 years, and histology of SCC were predictors; by multiple variable analysis, a short DFI (P = 0.018) and older age (P = 0.046) remained independent predictors of poor clinical outcomes. CONCLUSIONS: Young age and a long DFI are factors in favor of pulmonary metastasectomy after surgical treatment of primary head or neck cancers.

Spontaneous Regression of Metastatic Extraskeletal Myxoid Chondrosarcoma.

Spontaneous regression of tumors is very unusual and is defined as a partial or complete disappearance of metastatic tumors without any treatment. This phenomenon has been reported in almost all types of cancer. The patient was a 25-year-old woman who presented with multiple pulmonary nodules on her bilateral lungs on the annual chest roentgenograph. Simultaneously, a swelling mass on her subcutaneous inguinal region was observed. The diagnosis of the inguinal mass was extraskeletal myxoid chondrosarcoma. The inguinal mass and pulmonary nodules spontaneously regressed without any treatment after biopsy. The patient was doing well without evidence of recurrence at 1 year after the operation without any additional therapy. Our case is the first clinical one that indicated a possibility of histologic regression of extraskeletal myxoid chondrosarcoma.

Safety Evaluation of Hemoglobin-Albumin Cluster "HemoAct" as a Red Blood Cell Substitute.

A hemoglobin (Hb) wrapped covalently by human serum albumins (HSAs), a core-shell structured hemoglobin-albumin cluster designated as "HemoAct", is an O2-carrier designed for use as a red blood cell (RBC) substitute. This report describes the blood compatibility, hemodynamic response, and pharmacokinetic properties of HemoAct, and then explains its preclinical safety. Viscosity and blood cell counting measurements revealed that HemoAct has good compatibility with whole blood. Intravenous administration of HemoAct into anesthetized rats elicited no unfavorable increase in systemic blood pressure by vasoconstriction. The half-life of (125)I-labeled HemoAct in circulating blood is markedly longer than that of HSA. Serum biochemical tests conducted 7 days after HemoAct infusion yielded equivalent values to those observed in the control group with HSA. Histopathologic inspections of the vital organs revealed no marked abnormality in their tissues. All results indicate that HemoAct has sufficient preclinical safety as an alternative material for RBC transfusion.

Thirty-day outcomes after lobectomy or segmentectomy for lung cancer surgery.

BACKGROUND: Although lobectomy has been the standard operation for clinical T1aN0M0 non-small cell lung cancer, recent studies have shown that segmentectomy could be a substitute for lobectomy. However, the differences between lobectomy and segmentectomy in terms of perioperative complications have not been fully evaluated. METHODS: Between February 2006 and February 2013, 100 patients underwent lobectomy and 111 underwent segmentectomy for clinical T1aN0M0 non-small-cell lung cancer. We performed a retrospective comparison of perioperative morbidity, age, sex, pulmonary function, and Charlson comorbidity index in the 2 groups. Data were analyzed for all patients and for their propensity score-matched pairs. RESULTS: The incidence of postoperative complications in the segmentectomy group (n = 21, 19%) was significantly higher than that in the lobectomy group (n = 7, 7%; p < 0.01). The average operative time of 263 ± 64 min and estimated blood loss of 133 ± 125 mL for segmentectomy were significantly greater than those for lobectomy (201 ± 61 min and 88 ± 101 mL). In propensity score-matched analysis, the mean operative time of 270 ± 70 min for segmentectomy was longer than that for lobectomy (202 ± 67 min). Postoperative complications were more frequent in the segmentectomy group than in the lobectomy group (19.6% vs. 6.5%, p = 0.03). CONCLUSIONS: Although segmentectomy offers preservation of pulmonary function, significantly more postoperative complications occurred in the segmentectomy group. Surgeons should bear in mind that complications can happen more frequently after segmentectomy than after lobectomy for T1aN0M0 non-small-cell lung cancer.

Prognostic impact of preoperative tumor marker levels and lymphovascular invasion in pathological stage I adenocarcinoma and squamous cell carcinoma of the lung.

INTRODUCTION: Some unfavorable prognostic factors for stage I non-small-cell lung cancers have been reported; however, they are not reflected in the current Tumor-Node-Metastasis classification. METHODS: We retrospectively reviewed 629 patients who underwent complete resection of pathological stage I adenocarcinomas (ADs) or squamous cell carcinomas (SQs) at two institutes between 1996 and 2011. The correlation between clinicopathological characteristics and survival rates was analyzed to identify prognostic factors. RESULTS: Multivariate analysis indicated that among ADs, high serum carcinoembryonic antigen levels (p = 0.04 for overall survival [OS]; p < 0.01 for recurrence-free survival [RFS]; p = 0.02 for disease-specific survival [DSS]), lymphatic permeation (p < 0.01 for RFS and DSS), and vascular invasion (p < 0.01 for OS and RFS; p = 0.03 for DSS) were independent prognostic factors. Among SQs, high squamous cell carcinoma antigen (SCC) (p < 0.05 for OS), and vascular invasion (p < 0.05 for RFS and DSS) were independently prognostic. We suggest that among completely resected tumors less than or equal to 5 cm without lymph node metastasis, the current stages IA and IB AD with high serum carcinoembryonic antigen levels, lymphatic permeation, or vascular invasion should be upgraded to stage IB and IIA, respectively. The current stage IA SQ with high SCC antigen levels or vascular invasion should be upgraded to stage IB. These reclassifications accurately reflect survival status (p < 0.04 in all comparisons). CONCLUSIONS: Some important differences in prognostic factors were observed between AD and SQ. High preoperative serum tumor marker levels and lymphovascular invasion should be included as additional criteria in the forthcoming Tumor-Node-Metastasis staging.

Temporary extravascular shunt for reconstruction of a superior vena cava invaded by a lung tumor.

Advanced central lung cancers can invade the superior vena cava (SVC). Although the indications for resection of the vessel remain controversial, it has been suggested that it increases the long-term survival of selected patients; however, little consensus has been reached regarding the optimal method of vascular reconstruction. While the SVC is often replaced during unprotected cross-clamp, the placement of a temporary venous shunt with a view to preserve the periprocedural safety and facilitate the postoperative management seems preferable. We describe an SVC reconstruction procedure using an autologous pericardial patch and placement of a temporary extravascular shunt via a lateral thoracotomy.

Prognostic factors based on clinicopathological data among the patients with resected peripheral squamous cell carcinomas of the lung.

INTRODUCTION: Although the incidence of peripheral squamous cell carcinomas (p-SqCCs) of the lung has increased over recent years, clinicopathological factors influencing prognosis of resected p-SqCCs remain unclear. METHODS: We examined 280 patients who underwent complete resection of SqCCs and analyzed the clinicopathological features in relation to their overall survival (OS) and recurrence-free survival (RFS) according to the primary location. RESULTS: Multivariate analysis of all stages of p-SqCCs patients revealed that high serum squamous cell carcinoma antigen (SCC) level (OS; p < 0.01, RFS; p < 0.01), vascular invasion (OS; p < 0.01, RFS; p < 0.01), pleural invasion (OS; p = 0.03, RFS; p = 0.01), nodal metastasis (OS; p = 0.02) and complication with lung disease (OS; p < 0.01) were independently unfavorable prognostic factors. Among stage I p-SqCCs patients, high serum SCC level (OS; p < 0.01, RFS; p < 0.01), vascular invasion (RFS; p < 0.01) and pleural invasion (RFS; p = 0.01) were also strongly correlated with poor prognosis independently. When we reevaluated the survival rate, T1 p-SqCCs with high serum SCC level or vascular invasion can be upgraded to T2a. Patients with stage IB had a significantly poorer prognosis than stage IA (5-year RFS; 61.4 % versus 76.6 %, p < 0.05). CONCLUSION: High serum SCC level, pleural and vascular invasions were independent poor prognostic factors for completely resected p-SqCCs. T1 p-SqCCs with high serum SCC level or vascular invasion should be upgraded to T2a, which accurately reflect survival status among patients with p-SqCCs.

Surgical treatment of a solitary pulmonary metastasis from eyelid sebaceous carcinoma: report of a case.

BACKGROUND: Ocular sebaceous carcinoma is an uncommon, aggressive ocular neoplasm with potential for regional and distant metastasis. CASE PRESENTATION: A 77-year-old woman was found to have a solitary pulmonary lesion 6 years after the initial treatment of sebaceous carcinoma of the eyelid. Video-assisted lung wedge resection of an undetermined pulmonary nodule was carried out successfully. Microscopically, the tumor showed foamy cytoplasm and atypical nuclei, consistent with metastasis of eyelid sebaceous carcinoma. CONCLUSION: This is the first case report of resected solitary pulmonary metastasis of eyelid sebaceous carcinoma. Pulmonary resection is a good option for the treatment and diagnosis of metastatic eyelid sebaceous carcinoma.

[Resection and reconstruction of sternum].

We have experienced 6 cases with resection and reconstruction of sternum. They were 1 with osteosarcoma, 1 with synovial sarcoma, 1 with sternal metastasis of fallopian tube cancer, 1 with sternal metastasis of thyroid cancer, 1 with desmoid tumor, and 1 with dermatofibrosarcoma protuberance. Resection of both manubrium and sternum was performed in 3 cases and sternum resection in 3. There was no total resection. We used a titanium reconstruction plate and titanium mesh in 3 cases, a titanium reconstruction plate and polypropylene mesh in 2, titanium mesh in 1 for reconstruction of bony defect, and rectus abdominis myocutaneous flap in 3, pectralis major muscle flap in 2, latissimus doris myocutaneous flap in 1 for reconstruction of soft tissue defect. Postoperative courses were uneventful, and flail chest was not observed. Reconstruction of the bony defect of the anterior chest wall with the titanium reconstruction plate and titanium mesh or polypropylene mesh was effective by providing sufficient rigidity as well as protection of the thoracic organs.

A new technique of bronchial microsampling and proteomic analysis of epithelial lining fluid in a rat model of acute lung injury.

The standard technique to evaluate the proteins present in epithelial lining fluid (ELF) is bronchoalveolar lavage (BAL). Bronchoscopic microsampling (BMS) method has been developed for humans as a less invasive alternative. We establish the usefulness of a rat bronchial microsampling (rBMS) to evaluate various proteins in ELF in lipopolysaccharide (LPS)-induced lung injury models in rats. In the first experiment of this study, we validate that whether the rBMS can obtain information from ELF in place of BAL. Tumor necrosis factor (TNF)-α concentrations were increased in the rBMS samples similar to BAL 1 and 3h after LPS instillation. In the second part of this study, a proteomic analysis of the rBMS, using the Protein Chip(R) system, revealed the presence of proteins whose molecular weights corresponded to TNF-related proteins in the LPS-treated rats. In rats treated with a TNF-α converting enzyme inhibitor, the concentrations of these proteins in rBMS decreased or disappeared. In the third experiment, rBMS was performed without tracheostomy at 6 and 24h after instillation of LPS, and a rat multiple cytokines assay system detected heterogeneous variations in the concentrations of interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, TNF-αand interferon (IFN)-γ in ELF. The cytokine profile was significantly modified by pre-treatment with dexamethasone. This new rBMS technique could be used to measure TNF-α in LPS-induced acute lung injury (ALI) as well as for proteomic analysis, without sacrificing the rats. Furthermore, this procedure enables the serial collection of ELF, which would allow the examination of time-dependent cytokine variations in rat ALI model.