Adult ; Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; cytology ; Humans ; Macrophages ; pathology ; Male ; Middle Aged ; Pneumoconiosis ; pathology ; therapy

Animals ; Apoptosis ; Asthma ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Eosinophils ; cytology ; Guinea Pigs

OBJECTIVETo describe the pathologic features and diagnostic algorithm of pulmonary alveolar proteinosis (PAP).

METHODSThirty-nine biopsy and postmortem cases of PAP were studied by light microscopy and histochemical staining using periodic acid-Schiff (with digestion) (PAS-D), mucicarmine (with digestion) (mucicarmine-D) and alcian blue.

RESULTSHistologically, the affected lung tissue displayed the following characteristic features: (1) alveoli and some of the small bronchioles were filled with eosinophilic and fine granular proteinaceous material with needle-like clefts; (2) proteinaceous material was seen admixed with various numbers of degenerated and sometimes exfoliated pneumocytes; (3) pneumocytes were hyperplastic; (4) alveolar capillaries and alveolar septa had become hyperemic, but pulmonary interstitial inflammation was not obvious; (5) no significant inflammation was identified in the bronchial wall; (6) compensating emphysema was noted in the surrounding lung parenchyma. Fragments of eosinophilic, finely granular proteinaceous material with needle-like clefts were also found in the bronchoalveolar lavage fluid under light microscopy. The proteinaceous material was stained red by PAS-D. The staining for mucicarmine-D was negative, while alcian blue staining was either weakly positive (faint blue staining) or negative. Pathologic examination of lung biopsies and bronchoalveolar lavage fluid thus remaines the gold standard for diagnosis of PAP.

CONCLUSIONSIdentification of homogeneous, eosinophilic, finely granular and PAS-D-positive proteinaceous material with needle-like clefts in alveolar spaces or bronchoalveolar lavage fluid is of diagnostic importance in PAP. Bronchoalveolar lavage, being a relatively safe and non-invasive procedure, can be a useful adjunct in arriving at the final conclusion.

Adult ; Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; cytology ; Female ; Humans ; Lung ; pathology ; Male ; Middle Aged ; Periodic Acid-Schiff Reaction ; Pulmonary Alveolar Proteinosis ; pathology ; therapy ; Pulmonary Alveoli ; pathology

Bronchoalveolar Lavage Fluid ; Humans ; Interleukin-1beta ; metabolism ; Macrophages, Alveolar ; cytology ; drug effects ; metabolism ; Silicon Dioxide ; adverse effects ; Silicosis ; metabolism

To study the difference of changes on apoptosis, necrosis and respiratory burst of the polymorphonuclear neutrophils (PMN) in endotoxemia rat model. LPS (O(55)B(5), 5 mg/kg) was injected into abdominal cavity of 20 random normal Wistar rat. 2, 4, 8 and 12 hours after injection, the changes of apoptosis, necrosis and respiratory burst of the rats between PMN from the peripheral blood and from the bronchoalveolar lavage fluid were observed using the flow cytometer. At the same time, 5 uninjected rats were taken as control. The results demonstrated that the quantity proportions of apoptosis of PMN between the peripheral blood PMN and the bronchoalveolar lavage fluid PMN in rat's endotoxemia were similar. However, comparison with the uninjected LPS rat, the necrosis of peripheral blood PMN obviously increased and the respiratory burst capacity was clearly inhibited. Contrarily, the necrosis of bronchoalveolar lavage fluid PMN obviously decreased and the respiratory burst obviously increased in the injecting LPS rat. It was concluded that the necrosis and apoptosis displayed differently between the pulmonary and peripheral blood PMNs in endotoxemia. Under state of inflammation, the surviving PMN in tissue increased and kept the activated state due to tissue injury.
Animals ; Apoptosis ; Bronchoalveolar Lavage Fluid ; cytology ; Endotoxemia ; blood ; Necrosis ; Neutrophils ; physiology ; Pulmonary Alveoli ; pathology ; Rats ; Rats, Wistar ; Respiratory Burst

OBJECTIVETo investigate the effects of composite grinding dusts on rat respiratory system.

METHODSRats were administrated with grinding dusts by intratracheal injection. After 2 weeks, the total numbers of cells, the percentage of differential cell, the survival rate of cell, and the activity of lactate dehydrogenase(LDH) and alkaline phosphatase(ALP) in bronchoalveolar lavage fluid were analyzed.

RESULTSAlong with increasing concentration of grinding dusts, the total number of cells in lavage also increased, and was more than that in quartz group. Compared with control group, the percentage of neutrophil in lavage of rats treated with grinding dust and quartz significantly increased and meanwhile that of macrophage significantly decreased[PMN: quartz group (33.83 +/- 4.54)%; grinding dusts group (26.50 +/- 3.99)%, (36.00 +/- 3.58)%, (38.00 +/- 2.10)% at 10, 25, 50 mg/ml respectively. Macrophages: quartz group (62.17 +/- 4.54)%; grinding dusts group (70.83 +/- 3.66)%, (60.83 +/- 2.14)%, (58.17 +/- 2.48)%] while those in control group were (2.83 +/- 0.75)%, (95.67 +/- 1.21)% respectively. The cell survival rate in lavage in control group was 80%, but that in grinding dust and TiO2 group significantly decreased(P < 0.01). The activity of LDH and ALP in all rats treated with dusts obviously increased, and there was significant difference compared with control group(P < 0.01 or P < 0.05). Furthermore, there was significant difference between grinding dust group and quartz group, and between grinding dust group and TiO2 group respectively.

CONCLUSIONMetal grinding dust is very harmful to rat's lung cells and may cause fibrogenesis in the lungs.

Alkaline Phosphatase ; metabolism ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Dust ; L-Lactate Dehydrogenase ; metabolism ; Lung ; pathology ; Metals ; Rats

OBJECTIVETo evaluate the changes of CD(4)(+) IL-17+T (Th17) and CD(4)(+)Foxp3+regulatory T (Treg) cells in peripheral blood and bronchoalveolar lavage fluid (BALF) , and therefore to explore the role of Th17 and Treg in acrolein exposure airway inflammation in rats.

METHODSForty male Wistar rats were randomly divided into 4 groups: a 2 wk acrolein exposure group, a 4 wk acrolein exposure group, a 2 wk control group and a 4 wk control group (n=10 each). Cells in BALF were collected and analyzed by absolute and differential cell counts.IL-17 and IL-6 levels in serum and BALF were tested by enzyme linked immunosorbent assay (ELISA). The proportion of CD(4)(+)IL-17+T and CD(4)(+) Foxp3+Treg in peripheral blood and BALF were determined by flow cytometry.The mRNA expressions of IL-17 and Foxp3 were measured by real-time PCR. Comparisons of the data between different groups were performed using one-way ANOVA, and SNK and Games-Howell test were used for comparison between 2 groups.

RESULTSLevels of IL-17 were remarkable increased in the 2 wk acrolein exposure group and the 4 wk acrolein exposure group in serum [(52.64 ± 1.89) ng/L, (76.73 ± 5.57) ng/L], and BALF [(79.07 ± 5.67) ng/L, (96.61 ± 6.44) ng/L] compared with the 2 wk control group [(40.05 ± 3.12) ng/L, (56.75 ± 4.37) ng/L] and the 4 wk control group [(38.75 ± 3.23) ng/L, (53.27 ± 4.48) ng/L], all P<0.01. IL-6 was increased in the 2 wk and the 4 wk acrolein exposure group [ (33.28 ± 2.27) ng/L, (46.24 ± 3.16) ng/L] compared with the 2 wk and the 4 wk control group [ (16.37 ± 1.49) ng/L, (17.02 ± 1.43) ng/L] in BALF.Ratio of Th17 was higher in the 2 wk and the 4 wk acrolein exposure groups in peripheral blood (1.82 ± 0.18) %, (3.75 ± 0.48) % and BALF [(7.23 ± 0.27) %, (8.12 ± 0.38) %] compared with the 2 wk [(0.96 ± 0.07) %, (5.64 ± 0.63) %] and the 4 wk control group [(1.01 ± 0.08) %, (5.86 ± 0.57) %]. Ratio of Treg in BALF was higher in the acrolein exposure groups [ (8.83 ± 0.52) %, (12.05 ± 0.74) %] compared with the control groups [(4.37 ± 0.27) %, (5.01 ± 0.37) %]. The level of IL-17 mRNA was increased in the 2 wk and the 4 wk acrolein exposure group in peripheral blood [(25.78 ± 2.31), (34.69 ± 2.01) ] and in BALF [(23.04 ± 1.78), (34.56 ± 3.12)] compared with the 2 wk [(11.04 ± 2.53), (11.08 ± 2.05)] and the 4 wk [(12.03 ± 2.34), (12.69 ± 2.69)] control groups. Foxp3 mRNA was increased in the acrolein exposure groups [ (26.37 ± 3.24), (33.19 ± 2.98)] (24.4 ± 2.7), (30.3 ± 2.7) compared with the control groups [(12.37 ± 2.56), (13.12 ± 3.08)]. Th17 in acrolein exposure groups was positively correlated with counts of total cells and macrophages (r=0.5126, 0.5437, all P<0.01).

CONCLUSIONSA changed expression of Th17 and Treg cells and an vary of inflammatory cytokines were evident in airway inflammation of acrolein exposed rats, suggesting that Treg was involved in the immunological regulation and Th17 was associated with the persistent inflammation in acrolein induced airway inflammation in rats.

Acrolein ; toxicity ; Animals ; Bronchoalveolar Lavage Fluid ; cytology ; Cytokines ; metabolism ; Forkhead Transcription Factors ; metabolism ; Inflammation ; metabolism ; Male ; Rats ; Rats, Wistar ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

Animals ; Animals, Newborn ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Female ; Lipopolysaccharides ; toxicity ; Lung ; drug effects ; enzymology ; pathology ; Male ; Neutrophils ; cytology ; Peroxidase ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the effect of captopril on the histopathology and bronchoalveolar lavage fluid (BALF) in neonatal rats exposed to hyperoxia.

METHODSForty term neonatal Wistar rats were randomly assigned into Air control, Model, Normal saline control and Captopril-treated groups (n=10 each). The Air control group was exposed to air (FiO2=0.21). The remaining three groups were continuously exposed to hyperoxia (FiO2=0.90) . During exposure the Captopril-treated group received intragastric captopril (60 mg/kg daily) and the Normal saline control group was administered with normal saline. The Model group had no treatment. At the 14th and 21st days of exposure, the subjects were sacrificed. The lung coefficient and the protein contents and inflammatory cells in BALF were determined. The changes of lung histomorphology were observed.

RESULTSThe lung coefficient and the protein contents, the total number of cells and the percentage of neutrophils, lymphocytes and eosinophils in BAFL increased significantly in the Model and Normal saline control groups on the 14th and 21st days of exposure compared with those of the Air control group. Captopril treatment significantly reduced the lung coefficient and the protein contents, the total number of cells and the percentage of neutrophils and eosinophils in BALF. On the 14th day the lung coefficient decreased from 9.72 +/- 0.67 mg/g to 8.63 +/- 0.35 mg/g (P < 0.05); the protein contents in BALF from 0.619 +/- 0.023 g/L to 0.486 +/- 0.027 g/L (P < 0.05); and the total number of cells in BALF from (80.57 +/- 9.28)x10(4)/mL to (48.62 +/- 1.53)x10(4)/mL (P < 0.01) compared with the Model group. On the 21st day the lung coefficient decreased from 10.67 +/- 0.87 mg/g to 8.76 +/- 0.89 mg/g (P < 0.05); the protein contents in BALF from 0.978 +/- 0.012 g/L to 0.759 +/- 0.042 g/L (P < 0.05); and the total number of cells in BALF from (92.86 +/- 10.32) x10(4)/mL to (35.52 +/- 3.89) x10(4)/mL (P < 0.05) compared with the Model group. There were however significant differences in these results between the Captopril-treated and Air control groups. The histopathological examination demonstrated different degrees of alveolitis, broaden interstitium and reduced alveolar quantity in the Model and Normal saline control groups. The pathological changes were markedly alleviated after captopril treatment.

CONCLUSIONCaptopril may have protective effects on lung injury induced by hyperoxia.

Animals ; Animals, Newborn ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Captopril ; pharmacology ; Female ; Hyperoxia ; pathology ; Lung ; drug effects ; pathology ; Male ; Proteins ; analysis ; Rats ; Rats, Wistar

BACKGROUNDAsthma is a chronic airway disease with inflammation characterized by physiological changes (airway hyper-responsiveness, AHR) and pathological changes (inflammatory cells infiltration and mucus production). Eosinophils play a key role in the allergic inflammation. But the causative relationship between eosinophils and airway inflammation is hard to prove. One of the reasons is lack of activation marker of murine eosinophils. We investigated the expression of CD69 on murine eosinophils in vitro, the relationship between the expression of CD69 on eosinophils from peripheral blood and bronchoalveolar lavage fluid and on airway inflammation in asthmatic mice.

METHODSEosinophils from peripheral blood of IL-5 transgenic mice (NJ.1638) were purified. Mice were divided into five groups: wild type mice sensitized and challenged with saline (WS group), wild type mice sensitized and challenged with ovalbumin (WO group), IL-5(-/-) mice sensitized and challenged with saline and transferred with purified eosinophils (ISE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils (IOE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils, pretreated with anti CD4 monoclonal antibody (IOE+antiCD4mAb group). IL-5(-/-) mice were sensitized with OVA at day 0 and day 14, then challenged with OVA aerosol. On days 24, 25, 26 and 27 purified eosinophils were transferred intratracheally to IL-5(-/-) mice. On day 28, blood and BALF were collected and CD69 expression on eosinophils measured by flowcytometry.

RESULTSPurified eosinophils did not express CD69. But eosinophils cultured with PMA + MA, IFN-gamma, IL-5 or GM-CSF expressed CD69 strongly. Eosinophils from blood of WO, WS group did not express CD69 at all. The numbers of eosinophils in BALF of WO group, IOE group, ISE group and IOE + antiCD4mAb group were significantly higher than in mice of WS group which did not have eosinophils at all. CD69 expression on eosinophils in BALF of IOE and WO groups was strong. Eosinophils in BALF of ISE and IOE + antiCDmAb groups did not express CD69. The mucus production result was similar to CD69 expression. There were eosinophils infiltration in lung slides of all groups except WS group.

CONCLUSIONActivation in airway of eosinophils could directly lead to airway inflammation.

Animals ; Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; Asthma ; immunology ; physiopathology ; Bronchoalveolar Lavage Fluid ; cytology ; Eosinophils ; immunology ; Inflammation ; physiopathology ; Lectins, C-Type ; Lung ; physiopathology ; Mice ; Mice, Transgenic