Associação entre proteína 1 relacionada à uteroglobulina e gravidade da lesão por inalação de fumaça / Association of uteroglobin-related protein 1 with smoke inhalation injury severity

RESUMO Objetivo: Avaliar a expressão sérica da proteína 1 relacionada à uteroglobulina na fase inicial após lesões por inalação de fumaça e sua associação com a gravidade da lesão por inalação em pacientes queimados. Métodos: A lesão por inalação de fumaça ou produtos químicos se associa com morbidade e mortalidade. As consequências da inalação resultam de uma resposta inflamatória. A proteína 1 relacionada à uteroglobulina é anti-inflamatória e pode melhorar a inflamação pulmonar. Nossa hipótese é que os níveis de proteína 1 relacionada à uteroglobulina podem refletir a gravidade da doença e predizer o desfecho em pacientes com lesão por inalação. Incluíram-se prospectivamente neste estudo 16 pacientes com diagnóstico de síndrome do desconforto respiratório agudo decorrente de lesão por inalação de fumaça. Em todos os pacientes, colheu-se amostra de plasma quando da admissão à unidade de terapia intensiva, para avaliar a gravidade da lesão por inalação dentro de 72 horas. Os níveis plasmáticos de proteína 1 relacionada à uteroglobulina foram determinados em duplicata por meio de ensaio de imunoabsorção ligado à enzima. Resultados: A média de idade foi de 23 ± 5 anos, e a distribuição da lesão por inalação foi: três em grau 1, quatro em grau 2 e nove em grau 3. O nível de proteína 1 relacionada à uteroglobulina foi relacionado ao grau de severidade (grau 1: 0,389 ± 0,053 unidade arbitrária versus grau 2: 0,474 ± 0,0423 unidade arbitrária versus grau 3: 0,580 ± 0,094 unidade arbitrária; p = 0,007). Conclusão: Os níveis plasmáticos de proteína 1 relacionada à uteroglobulina se associam com o grau da lesão pulmonar por inalação.

ABSTRACT Objective: To evaluate serum uteroglobin-related protein 1 expression early after smoke inhalation injuries and its association with the severity of inhalation injury in burned patients. Methods: Smoke or chemical inhalation injury is associated with morbidity and mortality. The consequences of inhalation result from an inflammatory response. Uteroglobin-related protein 1 is an anti-inflammatory protein and may improve lung inflammation. We hypothesized that uteroglobin-related protein 1 levels could reflect disease severity and predict outcome in patients with inhalation injury. Sixteen patients diagnosed with acute respiratory distress syndrome secondary to smoke inhalation injury were prospectively included in the study. Plasma was collected upon intensive care unit admission and within 24 hours of the inhalation injury. Bronchoscopies were carried out in all patients to assess the severity of inhalation injury within 72 hours. Uteroglobin-related protein 1 plasma levels were determined in duplicate with enzyme-linked immunosorbent assay. Results: The mean age was 23 ± 5 years, and the inhalation injury distribution was as follows: three of grade 1, four of grade 2, and nine of grade 3. The level of uteroglobin-related protein 1 was related to inhalation severity (grade 1: 0.389 ± 0.053 arbitrary units versus grade 2: 0.474 ± 0.0423 arbitrary units versus grade 3: 0.580 ± 0.094 arbitrary units; p = 0.007). Conclusion: Plasma levels of uteroglobin-related protein 1 are associated with the degree of lung inhalation injury.

Dynamic changes in expression of clara cell protein and surfactant protein-D expressions in lung tissues and bronchoalveolar lavage fluid of silica-treated rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes in the expression of clara cell protein (CC16) and surfactant protein D (SP-D) in the lung tissues and bronchoalveolar lavage fluid (BALF) of silica-treated rats.</p><p><b>METHODS</b>Eighty-four Wistar rats were randomly divided into control group (n = 42) and silica group (n = 42). The silica group was subsequently divided into 3, 7, 14, 21, 28, and 60 d subgroups. The silicotic model was made by instilling silica suspension directly through the trachea into rat lungs. At 3, 7, 14, 21, 28, and 60 d after silica instillation, 8 rats in each group were sacrificed and their lung tissues and BALF were collected. The expression of SP-D and CC16 in lung tissues was detected by immunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of SP-D and CCl6 in BALF.</p><p><b>RESULTS</b>The immunohistochemical assay indicated that CCl6 and SP-D were expressed in lung cells. The ELISA found that in 7, 14, 21, 28, and 60 d silica subgroups, the content of CCl6 in rat BALF was 8.14±0.70, 7.15±0.66, 7.00±0.69, 6.34 ± 0.59, and 5.27±0.49 ng/L, respectively; CCl6 expression decreased gradually with the silica exposure time prolonged, indicating a negative correlation (ra = -0.953, P < 0.01). Compared with the control group, all silica subgroups had significantly decreased CCl6 levels (P < 0.05). The content of SP-D in BALF was 12.20 ± 1.57, 14.41 ± 0.65, and 12.18 ± 0.74 ng/L, respectively, in the 7, 14, and 21 d silica subgroups, significantly higher than that in the control group (P < 0.05).</p><p><b>CONCLUSION</b>The dynamic changes in SP-D and CCl6 protein levels in the lung tissues and BALF of rats could be induced by silica exposure and are related to silica exposure time. With the extension of silica exposure, CCl6 levels are gradually reduced, while the SP-D levels first increase and then fall.</p>

Bioinformatics of mouse uteroglobin binding protein and its polyclonal antibody preparation / 生理学报

To prepare anti-mouse uteroglobin binding protein (mUGBP) polyclonal antibody, two polypeptides were synthesized based on the bioinformatics analysis of mUGBP, and New Zealand white rabbits were immunized separately with each peptide coupled with keyhole limpet hemocyanin (KLH). The data indicate that a 13-amino acid polypeptide (positions 221st-233rd) was able to generate anti-peptide antibodies. The titer of the antisera detected with ELISA was 1:10(8). The antisera were then purified with immuno-affinity chromatography to obtain antibodies. Western blot analysis of mUGBP expressed as a fusion protein with a green fluorescent protein (GFP) was performed on the cell lysates of COS-1 cells with the purified antisera, suggesting that the antisera specifically recognized UGBP. By immunohistochemistry and indirect immunofluorescence analysis, we examined the expression of UGBP in the lung tissues from a patient undergoing surgical lung resection for a tumor and from normal mouse lung tissue, and found for the first time that UGBP protein was widely expressed in both mouse and human lung tissue with the most abundant expression in bronchial epithelial cells. These results suggest that the antigen epitopes of mUGBP are well predicted by using bioinformatics analysis. We have obtained anti-mUGBP polyclonal antibody, which will be useful for further investigation.

Effects of sevoflurane on pulmonary cytosolic phospholipase A₂ and clara cell secretory protein expressions in rabbits with one-lung ventilation-induced lung injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of sevoflurane on cytosolic phospholipase A₂ (C-PLA₂) and clara cell secretory protein (CCSP) in lung tissues of rabbits with one-lung ventilation (OLV)-induced lung injuries.</p><p><b>METHODS</b>Thirty-six healthy Japanese white rabbits were randomized into sham-operated group, OLV group, and OLV plus sevoflurane group subdivided into 4 subgroups with sevoflurane concentrations of 1%, 2%, 3% and 4%. CCSP and C-PLA₂ mRNA and protein expressions in rabbit lung tissues were detected by Western blotting and real-time PCR, and the content of arachidonic acid (AA) was measured using ELISA. The severities of the lung injury were evaluated according to lung wet/dry weight (W/D) ratio and histological scores.</p><p><b>RESULTS</b>In the OLV group and OLV+ sevoflurane groups, pulmonary CCSP expressions were significantly lower, while C-PLA₂ expression, lung W/D ratios and lung histological scores were significantly higher than those in the sham-operated group (P<0.05). Compared with OLV group, the OLV+sevoflurane groups showed significantly increased expressions of CCSP and reduced C-PLA₂ expression, lung W/D ratios and histological scores (P<0.05). In the 4 OLV+sevoflurane groups, CCSP expressions underwent no significant changes as sevoflurane concentration increased, but C-PLA₂ expressions, lung W/D ratios and histological scores all decreased gradually as the concentrations of sevoflurane increased (P<0.05).</p><p><b>CONCLUSION</b>OLV can result in down-regulated CCSP expressions and up-regulated C-PLA₂ expressions in rabbit lung tissues. Sevoflurane can protect against OLV-induced acute lung injury possibly by inhibiting C-PLA₂ expression via up-regulation of CCSP expressions or through other mechanisms resulting in down-regulated expression of C-PLA₂.</p>

Role of uteroglobin-binding protein in antiflammin-1 promoting IL-10 expression and secretion in RAW264.7 cells induced by endotoxin / 生理学报

The present study investigated the effect of antiflammin-1 (AF-1) on LPS-induced IL-10 secretion from RAW264.7 cells through uteroglobin-binding protein (UGBP). Cultured RAW264.7 cells, a murine monocyte-macrophage cell line, were divided as following: control group, LPS group (1 µg/mL LPS), AF-1 group (100 μmol/L AF-1), LPS+AF-1 group (2 h of 100 μmol/L AF-1 pretreatment before LPS addition), and LPS+AF-1+anti-UGBP group (30 min of anti-UGBP antibody pretreatment before successive treatments with AF-1 and LPS). IL-10 concentration in the supernatants was detected by ELISA assay, and the level of IL-10 mRNA expression in macrophage was detected by using RT-PCR method. The results showed that AF-1 significantly increased LPS-induced IL-10 secretion in RAW264.7 cells in a dose dependent way, and up-regulated its mRNA level. Anti-UGBP antibody pretreatment attenuated the augmented effect of AF-1 on LPS-induced IL-10 secretion and gene expression. These results suggest that AF-1 promotes LPS-induced IL-10 secretion from macrophages, and this effect is mediated by UGBP.

Regulation on the expression of Clara cell secretory protein in the lungs of the rats with acute lung injury by growth hormone / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Clara cell secretory protein (CC16) is an important lung derived protective factor and may play an important role on the pathogenesis of acute lung injury (ALI) induced by endotoxemia. Growth hormone (GH) is an important anabolism hormone secreted by GH cells of the hypophysis. Previous research showed that GH would significantly exacerbate ALI induced by endotoxemia, but the mechanism is not very clear yet. Whether the effects are related to CC16 or not is undetermined.</p><p><b>METHODS</b>One hundred and twelve male Sprague-Dawley rats were randomly divided into an ALI group and a GH group. The rats in the two groups were subdivided into seven subgroups, according to injection with lipopolysaccharides (LPS) or not, then according to different intervals of time after LPS injection; 0 hour (pre-injection of LPS, acted as control group), 0.5 hour, 1 hour, 2 hours, 4 hours, 6 hours and 24 hours for subgroups. Pulmonary alveolar septa area density (PASAD) and ploymorphonuclear cells (PMN) in the lungs were analyzed morphometrically. The levels of tumor necrosis factor (TNF) and interleukin 6 (IL-6) were determined by radioimmunoassay. To analyze the expression and activation of nuclear factor kappa B (NF-κB), the numbers of NF-κB positive cells in lungs were counted after immunofluorescence staining, and the levels of NF-κB inhibitory protein-α (IκB-α) in lung homogenates of rats were detected by Western blotting. The expression levels of CC16 mRNA in lungs of the rats with ALI were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The levels of CC16 protein in lung homogenates were detected by Western blotting.</p><p><b>RESULTS</b>Half an hour after LPS injury both the PASAD and PMN numbers in lungs of the rats with ALI began to increase significantly and peaked at 6-hour post-injury. They then began to recover and reached normal levels at 24-hour post-injury. Both the PASAD and PMN numbers in the GH group increased more significantly than those in the ALI group. The levels of TNF in lungs of the rats with ALI homogenates increased significantly 0.5-hour post-injury, peaked at 1-hour and maintained a high level until 6 hours then gradually recovered. The content of TNF in the GH group lung homogenates increased more significantly than in the ALI group post-injury. The contents of IL-6 in rat lung homogenates began to increase significantly at 1-hour post-injury, peaked at 4 hours then gradually returned to normal levels by 6 hours post-injury. The levels of IL-6 in the lung homogenates of the GH group were higher than in the ALI group at different time intervals post-injury. The number of NF-κB positive cells increased dramatically at 0.5-hour post-injury, and the fluorescence intensity was enhanced. Both peaked at 4-hour post-injury. The number of NF-κB positive cells and the enhanced intensity of fluorescence began to decrease from 6-hour post-injury, but the number of NF-κB cells at 24 hours post-injury was still higher than in the control group. The number of NF-κB cells in lungs in the GH group was significantly higher than in the LPS group at the different time intervals post-injury. The IκB-α expression in lungs of the rats with ALI homogenates decreased dramatically 0.5-hour post-injury, reaching a nadir at 4-hour post-injury and then began to recover. The levels of IκB-α in GH group were significantly lower than those in ALI group. Both the levels of CC16 mRNA and protein in lungs of the rats with ALI began to decrease significantly 0.5-hour post-injury, reached a nadir at 6 hours, and then began to recover. Both the expression of CC16 mRNA and CC16 protein in the GH group were significantly lower than those in the ALI group at the different time intervals post-injury. Correlation analysis indicates that CC16 correlates significantly with all the indices mentioned above.</p><p><b>CONCLUSIONS</b>Down-regulation of CC16 expression plays a critical role in the pathogenesis of acute lung injury induced by endotoxemia. The application of GH can exacerbate the lung injury induced by endotoxemia through down-regulating the expression of CC16.</p>

Neutrophil predominance in induced sputum from asthmatic patients: Therapeutic implications and role of clara cell 16-KD protein / Predominio de neutrófilos en esputo inducido de asmáticos: Implicancias terapéuticas y rol de la proteína 16-kD de la célula clara

Eosinophil is considered to be a main protagonist in asthma; however, often discordances between clinical manifestations and response to treatment are observed. We aimed to determine the occurrence of neutrophil predominance in asthma and to identify its characteristics on the basis of clinical-functional features, induced sputum cellular pattern and soluble molecules, to guide the appropriated anti-inflammatory therapy. A total of 41 patients were included in randomized groups: 21-40 year-old, with stable mild-to-severe asthma, steroid-naïve and non-smokers. An induced sputum sample was obtained under basal conditions, a second one after treatment with budesonide (400 µg b.i.d.) or montelukast (10 mg/d) for six weeks, and a final one after a 4-week washout period. By cytospin we evaluated eosinophil (EP) or neutrophil predominance (NP), and in supernatant we determined LTE4, and CC16. Peak expiratory flow variability (PEFV) was measured. A total of 23/41 patients corresponded to EP and 18/41 patients to NP. The PEFV was higher in EP than in NP. LTE4 was higher with NP than with EP. No difference was found for CC16. Montelukast reduced the predominant cell in both subsets, whereas budesonide only reduced eosinophils in EP. Budesonide and montelukast reduced PEFV in EP but not in NP. Considering the total treated-samples in each subset, CC16 level increased significantly in EP. In conclusion: a NP subset of asthmatic patients was identified. These patients show a lower bronchial lability; the leukotriene pathway is involved which responds to anti-leukotriene treatment. This phenotype shows a poor recovery of CC16 level after treatment.

El eosinófilo es considerado la célula protagonista principal en el asma; sin embargo, a menudo se observan discordancias entre las manifestaciones clínicas y la respuesta de los pacientes al tratamiento. Nos propusimos determinar la ocurrencia de predominio de neutrófilos en el asma e identificar las características clínico-funcionales, el patrón celular y las moléculas solubles del esputo inducido, para guiar el tratamiento apropiado anti-inflamatorio. Se incluyeron 41 pacientes: 21 a 40 años de edad, con asma estable leve a grave, no tratados con esteroides tópicos ni sistémicos y no fumadores. Se obtuvo una muestra de esputo inducido en condiciones basales, una segunda muestra después del tratamiento al azar con budesonida (400 µg dos veces al día) o el montelukast (10 mg/d) durante seis semanas, y una final después de un período de lavado de 4 semanas. En el frotis por citocentrifugado se evaluó el predominio de eosinófilos (EP) o neutrófilos (NP), y en el sobrenadante se determinó LTE4, y CC16. Se midió la variabilidad del flujo espiratorio máximo (PEFV). Un total de 23/41 pacientes correspondieron al EP y 18/41 pacientes con NP. El PEFV fue mayor en el EP que en NP. LTE4 fue mayor en NP que en EP. No se encontraron diferencias de los niveles de CC16 en ambos grupos. Montelukast redujo la célula predominante en ambos subgrupos, mientras que budesonida sólo redujo los eosinófilos en EP. Tanto budesonida como montelukast redujeron PEFV en EP, pero no en NP. El nivel de CC16 aumentó significativamente en el EP luego del tratamiento antiinflamatorio. En conclusión: se identificó un subgrupo de asmáticos NP que presentan una menor labilidad bronquial, la vía de los leucotrienos parece estar involucrada y responde al tratamiento anti-leucotrienos. Este fenotipo muestra una escasa recuperación del nivel de CC16 posterior al tratamiento.

Expression of human mammaglobin and clinicopathologic correlations in breast cancer: The findings in Malaysia.

Background: Human mammaglobin (hMAG) is a secreted protein which has been detected in breast epithelial cells of mammary glands and has been used as a specific marker for breast cancer. Objectives: This study aims at studying the hMAG expression and identifying the significant predictors of hMAG expression in breast cancer tissues. Materials and Methods: The tissue samples were obtained from two major teaching hospitals in the country. They were examined by immunohistochemistry (IHC) and the hMAG expression was evaluated using an established scoring system. Results: Out of 84 breast cancer tissue samples, hMAG was expressed in 50 samples (59.6%). The expression of hMAG was found to be increased with cancer grade. The output of logistic regression model showed that hMAG was overexpressed in breast cancer samples from the first hospital (P = 0.014), but not with those from the second hospital. Conclusions: It can be concluded that hMAG may serve in the diagnosis and the assessment of progression with the increased cancer grade. The dominance in hMAG expression in samples from HUSM may correlate with ethnic, environmental or genetic factors.

Expression of serum Clara cell secretory protein 10 in wheezing children under 5 years old / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the expression of serum Clara cell secretory protein 10 (CC10) and total IgE concentration in wheezing children under 5 years old.</p><p><b>METHODS</b>Fifty-nine children with recurrent wheezing under 5 years old were classified into two groups: wheezing group 1 with atopic high risks (n=33) and wheezing group 2 without atopic high risks (n=26). Twenty-three children without infectious diseases served as a control group. Serum levels of CC10 and IgE were measured using a solid-phase sandwich ELISA.</p><p><b>RESULTS</b>The serum levels of CC10 in wheezing group 1 (3.95 ± 1.26 ng/mL) and wheezing group 2 (5.41 ± 1.64 ng/mL) were significantly lower than those in the control group (8.72 ± 2.23 ng/mL; P<0.01). The wheezing group 1 showed more decreased serum levels of CC10 compared with wheezing group 2 (P<0.05). The serum IgE levels in wheezing group 1 were significantly higher than those in wheezing group 2 and the control group (P<0.05). There were no significant differences in serum IgE levels between the wheezing group 2 and control group. There was a negative correlation between serum levels of CC10 and IgE in wheezing group 1 (r=-0.912, P < 0.01).</p><p><b>CONCLUSIONS</b>Serum CC10 levels decrease remarkably in wheezing children, and more significant decrease is noted in patients with atopic high risks. Serum CC10 levels are negatively correlated to serum IgE levels in patients with atopic high risks.</p>

Effects of N-acetylcysteine on Clara cells in rats with cigarette smoke exposure / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The number of Clara cells and the Clara cell 16-kDa protein (CC16) levels of the lung decrease in patients with chronic obstructive pulmonary disease (COPD). N-acetylcysteine (NAC) is a powerful antioxidant and can reduce the frequency of acute exacerbations of COPD. But the exact mechanism is unclear. The present study was designed to investigate the effects of NAC on Clara cells in rats with cigarette smoke exposure.</p><p><b>METHODS</b>Eighteen adult male Wistar rats were randomly divided into 3 groups, 12 exposed to cigarette smoke (CS) thrice a day, 10 cigarettes for 30 minutes each time for 1 week, without (CS group) or with (CS + NAC group) oral intake of NAC 80 mg x kg(-1) x d(-1), and another 6 rats exposed to fresh air (control group). Clara cells were observed by an electron microscope. The mRNA expression of CC16 and CC16 protein in lungs were determined by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry respectively. The glutathion (GSH) level in plasma and lung tissue were tested by fluorimetry assay.</p><p><b>RESULTS</b>Compared with the controls, the pathologic score of small airways significantly increased in the CS exposed rats (20.3 +/- 14.7 vs. 53.7 +/- 11.5, P < 0.05). The Clara cell particles in cytoplasm decreased in the CS group (P < 0.05). The percentage of CC16-positive cells in bronchioles in the CS group (27.8 +/- 4.3 and 29.5 +/- 2.4 in terminal bronchioles and respiratory bronchioles, respectively) significantly decreased as compared with the control group (37.1 +/- 3.8 and 43.8 +/- 5.8 in terminal bronchioles and respiratory bronchioles, respectively) (P < 0.05). No significant difference was observed in GSH level ((181 +/- 26) nmol/L in the control group vs. (170 +/- 18) nmol/L in the CS group) between the two groups. After treatment with NAC, the pathologic score of small airways (24.1 +/- 17.5) decreased (P < 0.05). Clara cell particles in cytoplasm of Clara cells increased and GSH level in plasma ((213 +/- 40) nmol/L vs. (170 +/- 18) nmol/L in the CS group) increased too (P < 0.05), while the increase in the proportions of CC16 positive cells in bronchioles (30.1 +/- 6.4 and 34.3 +/- 6.3 in terminal bronchioles and respiratory bronchioles, respectively) did not reach the statistical significance (P > 0.05). No significant difference was found in the expression of CC16 mRNA among the three groups. Correlation analysis indicated that the percentage of CC16-positive cells in bronchioles negatively correlated with the pathologic score of small airways (r = -0.592, P < 0.05), but not with GSH level.</p><p><b>CONCLUSIONS</b>One-week CS exposure decreased the number of Clara cells and the expression of CC16 in bronchioles in rats. NAC might provide protection of the Clara cells from oxidative damage and possibly through the elevation of the synthesis and secretion of CC16. These data indicate that NAC decreases airway inflammation induced by CS via induction of CC16.</p>

Expression and significance of Clara cell secretory protein in injury lungs of Kunming mice after n-hexane long-term inhalation / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the expression of Clara cell secretory protein(CCSP) in the Kunming mouse model of n-hexane long-term inhalation, and to discuss the functions of Clara cell in injury lung induced by n-hexane.</p><p><b>METHODS</b>24 healthy mice were randomly divided into 4 groups: one control group and three n-hexane groups (4 w, 8 w and 12 w), 6 each group. Primary concentration of n-hexane was 17.6 g/m3, 8 hours per day, 6 d per week. After inhalation, n-hexane concentration of blood from celiac artery was detected. The lungs were embedded with paraffin and HE staining in the routine. The ratio of Clara cells with CCSP reaction in bronchiole and the number of macrophage cells with lysozyme reaction were determined by immuno-histochemistry.</p><p><b>RESULTS</b>In the poisoning groups, the average n-hexane concentration of blood was significantly higher than that of the control group (P < 0.01). There were apparent pathologic damages in lungs of the poisoning mice. In poisoning 4 w, 8 w and 12 w groups, the ratio of Clara cells was significantly decreased [(73.33 +/- 4.21)%, (60.98 +/- 4.94)%, (34.04 +/- 2.33)% in terminal bronchiole, and (75.44 +/- 7.91)%, (58.54 +/- 4.86)%, (33.35 +/- 2.67)% in respiratory bronchiole] as compared with the control mice [(80.26 +/- 6.43)% and (81.74 +/- 7.75)%, P < 0.05 or P < 0.01], meanwhile the numbers of macrophage cells were gradually increased [(21.39 +/- 7.41), (28.54 +/- 10.73), (48.97 +/- 19.55) per microscopic field at 200x] in poisoning mice than those in control mice [(7.84 +/- 3.12) per microscopic field at 200x, P < 0.05 or P < 0.01].</p><p><b>CONCLUSION</b>In injury lungs after n-hexane inhalation, Clara cells are the target cells of n-hexane toxicity effect. Clara cells play an extensive protective role in lung inflammation.</p>

Evaluation of intraoperative molecular assay in sentinel lymph node biopsy from breast cancer patients / 中华病理学杂志

<p><b>OBJECTIVE</b>to evaluate the application of GeneSearch(TM) breast lymph node assay in intraoperative detection of metastases in sentinel lymph node (SLN) from breast cancer patients.</p><p><b>METHODS</b>a total of 225 SLN from 88 patients was prospectively studied. Each SLN was cut into 2 mm slabs which were examined by intraoperative imprint cytology (IIC) first, followed by GeneSearch assay and post-operative serial sectioning. GeneSearch used real-time fluorescence quantitative RT-PCR technology to detect the expression of CK19 and mammaglobin in SLN. The results of GeneSearch assay were correlated with those of IIC and post-operative serial sectioning.</p><p><b>RESULTS</b>amongst the 88 cases studied, 225 SLNs were found, and obvious metastatic carcinoma cells were identified in 27 SLNs and micrometastasis in 9 SLNs. One hundred and eight-nine SLNs were considered as "negative" (with "isolated tumor cells" present in 5 SLNs). The turn-around time of intraoperative GeneSearch assay ranged from 35 to 45 minutes (mean = 40 minutes). The concordance rate between GeneSearch assay and post-operative serial sectioning was 95.6% (215/225), with a sensitivity of 86.1% (31/36), compared with 94.7% (213/225) and 72.2% (26/36) respectively for IIC. The size of metastatic foci correlated with the Ct value of CK19 and mammaglobin (P < 0.01).</p><p><b>CONCLUSIONS</b>GeneSearch assay for intraoperative detection of metastase in SLN has a satisfactory performance and demonstrates a relatively higher sensitivity than IIC. The potential clinical application still requires further evaluation of larger number of cases.</p>

Expression of COX-2 and IDO by Uteroglobin Transduction in NSCLC Cell Lines / 결핵및호흡기질환

BACKGROUND: Uteroglobin (UG) is a secretary protein that has strong immunomodulatory properties, and which is synthesized in most epithelia including lung tissue. Overexpression of UG is associated with decreased expression of cyclooxygenase (COX)-2 and suppression of cancer cell growth. Indoleamine 2,3-dioxygenase (IDO) catalyzes tryptophan along the kynurenine pathway, and both the reduction in local tryptophan and the production of tryptophan metabolites contribute to the immunosuppressive effects of IDO. METHODS: In this study, we investigated the pattern of expression of COX-2 and IDO, and the effect of UG transduction in the expression of COX-2 and IDO in several non-small cell lung cancer cell lines, especially A549. RESULTS: Both COX-2 and IDO were constitutionally expressed in A549 and H460 cells, and was reduced by UG transduction. In A549 cells, the slightly increased expression of COX-2 and IDO with the instillation of interferon-gamma (IFN-gamma) was reduced by UG transduction. However, the reduced expression of COX-2 and IDO by UG transduction was not increased with IFN-gamma instillation in A549 cells. In both the A549 COX-2 sense and the A549 COX-2 anti-sense small interfering RNA (siRNA)-transfected cells, IDO was expressed; expression was reduced by UG transduction, irrespective of the expression of COX-2. CONCLUSION: The results suggest that the anti-proliferative function of UG may be associated with the immune tolerance pathway of IDO, which is independent of the COX-2 pathway.

Lung adenocarcinoma stem cell phenotypes and their correlation with patient prognosis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the cancer stem cells and to evaluate their prognostic implication in patients with lung adenocarcinoma.</p><p><b>METHODS</b>Three phenotypic markers of cancer stem cells (SP-C, CCSP and OCT4) in lung adenocarcinoma were detected by immunofluorecence staining. The correlation among the clinicopathological parameters and phenotypes of cancer stem cells as well as survival were analyzed by Cox proportional hazard method.</p><p><b>RESULTS</b>Of the 57 cases, cancer stem cells were detected in 52, including OCT4(+) bronchioloalveolar stem cell (BASC) phenotype (SP-C(+) CCSP(+) OCT4(+)) in 40 cases and OCT4(-) BASC phenotype (SP-C(+) CCSP(+) OCT4(-)) in 12 cases. Statistical analysis revealed that the phenotype of cancer stem cells was related with the cellular differentiation, i.e. the OCT4(+) BASC phenotype occurred more frequently in the well-differentiated tumors, while the OCT4(-) BASC phenotype usually presented in most of the poorly-differentiated ones. Cox analysis showed that the OCT4(+) BASC phenotype was one of prognostic factors.</p><p><b>CONCLUSION</b>The lung adenocarcinoma stem cells have phenotypic features of bronchioalveolar stem cells (SP-C(+) CCSP(+)). The expression of self-renewal regulatory gene OCT4 in these cells indicates an aggressive nature and unfavorable prognosis.</p>

Expression of clara cell 10-KDa protein in murine bacterial chronic rhinosinusitis model / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression of Clara cell 10-KDa protein (CC010) in sinonasal mucosa of murine bacterial chronic rhinosinusitis (BCRS) model.@*METHOD@#A murine BCRS model was established by Streptococcus pneumoniae inoculation plus Merocel ostiomeatal obstruction. After 12 week's intervention, histological changes of sinonasal mucosa in BCRS model were examined by hematoxylin and eosin stain, periodic acid-schiff stain, and Masson-Trichrome stain. The mRNA and protein expression of CC10 in sinonasal mucosa were determined by reverse transcription polymerase chain reaction and immunohistochemistry methods. The number of CC10 positive cells in sinonasal epithelium was also counted.@*RESULT@#In BCRS model group, polymorphonuclear neutrophils (PMN), subepithelial collagen deposition, goblet cells, and epithelial thickness were significantly increased, compared with control group (P<0.01). However, CC10 positive cells, CC10 mRNA and protein expression in sinonasal mucosa of BCRS model group were significantly decreased, compared with control group (P<0.01). Moreover, the number of CC10 positive cells was significantly negatively correlated with PMN (r=-0.734, P<0.01), subepithelial collagen deposition (r=-0.776, P<0.01), epithelial goblet cells (r=-0.841, P<0.01), and epithelial thickness (r=-0.805, P<0.01), respectively. CC10 average grayscale value was significantly positively correlated with PMN (r=0.771, P<0.01), subepithelial collagen deposition (r=0.802, P<0.01), epithelial goblet cells (r=0.887, P<0.01), and epithelial thickness (r=0.855, P<0.01), respectively.@*CONCLUSION@#The expression of CC10 is downregulated in sinonasal mucosa in BCRS model. As an important endogenous modulin, CC10 might play a crucial role in the pathogenesis of chronic rhinosinusitis.

Dynamic changes of surfactant protein-D and Clara cell protein expressions in lung tissue and bronchoalveolar lavage fluid of silica-treated rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes of surfactant protein-D (SP-D) and Clara cell protein (CC16) expressions in lung tissue and bronchoalveolar lavage fluid (BALF) of silica-treated rats.</p><p><b>METHODS</b>80 rats were randomly divided into the control group and the silica group. The silicotic animal model was established by direct tracheal instillation of silica suspension into rat lungs surgically. On 7, 14, 21, 28 and 60 d after establishment of the animal model, eight rats in each group were sacrificed and lung tissue and BALF were collected. Lung tissue chip microarray was made in different time points after the silica was injected. Expressions of SP-D and CC16 on tissue microarray were detected with immunohistochemistry and quantified by Image-Pro Plus Version 4.5 for Windows(TM); The SP-D and CC16 levels of BALF were detected with western blot and quantified by Quantity One Version 4.6.2.</p><p><b>RESULTS</b>SP-D expressed very little in alveolar type II and Clara cell intracytoplasmic of control group while its expression significantly increased after 7 d in silica group (P < 0.01) and it reached the peak on the 14 d, after this SP-D expression decreased gradually. CC16 was expressed strongly in intracytoplasmic and it expressed little in nucleus of Clara cell by bronchioles of control while it significantly decreased after 7 d in silica group (P < 0.01), and CC16 expression decreased gradually with the exposed silica time, which was correlated negatively among them (r(s) = -0.967, P < 0.01). On 7 d and 28 d, the SP-D levels of BALF in silica group were significantly higher than control (P < 0.01). Furthermore the SP-D levels of BALF on 28 d was significantly elevated than that on 7 d in silica group (P < 0.01). On 7 d and 28 d, the CC16 levels of BALF in silica group were significantly lower than control (P < 0.01). Moreover, CC16 levels of BALF on 28 d was significantly decreased than that on 7 d in silica group (P < 0.01).</p><p><b>CONCLUSION</b>The dynamic changes of SP-D and CC16 protein expressed in lung tissue and bronchoalveolar lavage fluid could be induced by silica exposure and are related with the silica exposure time.</p>

Changes of Clara cell protein and surfactant protein-D in serum of patients with silicosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore changes of Clara cell protein (CC16) and surfactant protein-D (SP-D) in the serum of patients with silicosis.</p><p><b>METHOD</b>The concentrations of CC16 and SP-D were measured in the serum by sandwich enzyme-linked immunosorbent assays. The subjects consisted of 30 healthy volunteers and 90 silica-exposed workers including silica-exposed group, the silicosis of suspects group (0(+)) and the silicosis phase I group, 30 subjects each groups.</p><p><b>RESULTS</b>The concentrations of CC16 in the serum was significantly decreased in silica-exposed workers compared to controls (P < 0.01); The concentrations of CC16 in the serum were higher in lifelong nonsmokers than the current smokers in control subjects (P < 0.05), but they were no differences between lifelong nonsmokers and current smokers of 90 silica-exposed workers. Compared with control subjects, the levels of SP-D in the serum of silicosis suspects (0(+)) and silicosis phase I groups were significantly elevated (P < 0.01, respectively), which were also higher than silica-exposed group (P < 0.05 and P < 0.01, respectively), Discriminant equations set by CC16 and SP-D were used in diagnosis of silicosis, and the rate of accuracy in healthy volunteers, the silica-exposed group and the silicosis phase I group were 86.7%, 86.7% and 76.7%, respectively, The total rate of correct classification hit 84.2%.</p><p><b>CONCLUSION</b>The serum CC16 of long-term silica-exposed workers is decreased, and SP-D is increased gradually.</p>

Effects of recombinant human insulin-like growth factor-1 on the expression of Clara cell secretory protein in lung of hyperoxia-exposed newborn rats / 中华儿科杂志

<p><b>OBJECTIVE</b>The development of neonatology and the availability of pulmonary surfactant have been helpful in effective reduction of the mortality of very low birth weight infants at the expense of an increasing number of survivors with bronchopulmonary dysplasia (BPD) caused by lung immaturity. BPD is a common syndrome in newborns, especially in preterm infants, when treated with hyperoxia and mechanical ventilation. Unfortunately, there have been no effective measure for the prevention and treatment of BPD. The purpose of this study was to investigate the influence of recombinant human insulin-like growth factor-1 (rh-IGF-1) on cell apoptosis and Clara cell secretory protein (CCSP) expression during the lung injury induced by hyperoxia, so as to assess its effect on the inflammatory lung injury and its developmental repair.</p><p><b>METHODS</b>Eighty full term neonatal Wistar rats under the same condition were divided randomly into four groups on the second day after birth. Group I was air control, group II was exposed to hyperoxia, group III air + rh-IGF-1, and group IV was treated with hyperoxia + rh-IGF-1. The pups in the control group were kept in room air, while pups in hyperoxia group were kept in a Plexiglas chamber and exposed to over 85% oxygen. Pups in group III were under the same raising condition except for exposure to room air and treated with intraperitoneal injection of rh-IGF-1 (1 microg/Kg) everyday from the third day. Pups in group IV were treated with intraperitoneal injection of rh-IGF-1 (1 microg/Kg) everyday from the third day of exposure to hyperoxia. Lung tissue sections of the neonatal rats were stained with hematoxylin and eosin (HE) after 7 d of hyperoxia exposure, expression of CCSP was examined by immunohistochemical method, and apoptotic cell index of lung tissue was calculated by using TUNEL method.</p><p><b>RESULTS</b>It was observed from immunohistochemical examination that positive staining of CCSP was distributed mainly in distal and respiratory bronchioles. The percentage of Clara cells in distal and respiratory bronchioles epithelium decreased in hyperoxia group (32.17 +/- 3.19)% compared to that in air control group (68.32 +/- 2.04)%, P < 0.01. Statistically significant differences were found in intensity of positiveness of Clara cells between hyperoxia (29.45 +/- 5.56) and air control group (42.37 +/- 3.24), P < 0.01. TUNEL assay showed that most apoptotic cells were alveolar and bronchial epithelial cells. The apoptotic index increased significantly in the hyperoxia group (55.77 +/- 6.09)% compared to the air control group (16.41 +/- 4.01)%, (P < 0.01). The positive rate (52.98 +/- 2.68)% of Clara cells and the expression (41.22 +/- 6.36) of CCSP in hyperoxia + rh-IGF-1 group increased significantly when compared with hyperoxia group, and the differences between these two group were also statistically significant (P < 0.01). The apoptotic index increased significantly in the hyperoxia + rh-IGF-1 group (27.98 +/- 3.09)% compared to the hyperoxia group (P < 0.01).</p><p><b>CONCLUSIONS</b>Hyperoxia exposure can promote the pneumocyte apoptosis and inhibit the expression of CCSP. Rh-IGF-1 can remove the block of the formation of lung alveoli, increase the secretion of CCSP, mitigate inflammatory responses in airway and alleviate lung injury via pneumocyte apoptosis. Therefore, the results of this study provide a theoretic and experimental evidence for clinical application of rh-IGF-1 in prevention and treatment of BPD.</p>

Combining mammaglobin and carcinoembryonic mRNA markers for early detection of micrometastases from breast cancers--a molecular study of 59 patients.

INTRODUCTION: As many as 30% of node-negative breast cancer patients relapse within five years, suggesting that current histological detection methods are inadequate for identifying metastatic disease. Detecting small number of cancer cells in the breast tissue or lymph node by reverse transcription-polymerase chain reaction (RT-PCR) assays using a combination of tissue and cancer specific markers might be very useful in the early detection or monitoring of the treatment. Mammaglobin is a member of the uteroglobin gene family and appears to be expressed only in breast tissue. Carcinoembryonic antigen has been the preferred molecular marker for detection of micro metastases in lymph nodes in almost all carcinomas. MATERIALS AND METHODS: Samples were collected from randomly chosen breast cancer patients undergoing modified mastectomy or breast conserving surgery between September 2003 and July 2004. RT-PCR was applied to study the expression of MMG and CEA markers. Breast cancer micrometastases in axillary lymph nodes were also assessed. RESULTS: The MMG marker was positive in 9/10 normal breast tissues, 3/3 breast fibroadenomas and 37/39 of breast carcinoma tissues, giving an overall sensitivity of 94%. The sensitivity was 80% for metastatic lymph node samples. On the other hand CEA showed 95% sensitivity for malignant breast tumors and 100% sensitivity for metastatic lymph nodes. CONCLUSIONS: RT-PCR using a combination of MMG and CEA markers is a powerful tool to complement current routine histopathology techniques for detection of breast cancer metastasis in axillary nodes.