ObjectiveTo investigate the changes of autophage-related protein in lung tissues of rats with chronic obstructive pulmonary disease (COPD). MethodsPassive cigarette smoking was used to establish COPD model in rats. The mRNA and protein expressions of PI3K, total AKT, phosphorylated-AKT, total mTOR, phosphorylated-mTOR, and autophagy-related genes including LC3Ⅱ/Ⅰ, Beclin1, Atg5, Atg7, Atg12, P62 in lung tissues were measured by real-time PCR and Western blot. The autophagy level was compared between the COPD rats and the normal rats by LC3B immunohistochemistry. ResultsReal-time PCR analysis showed that the mRNA expressions of Beclin1, Atg5 and Atg12 significantly increased in lung tissues of the COPD rats compared with the normal rats (all P < 05). There was no significant difference between the COPD rats and the normal rats as for Atg7 mRNA expression (P > 0.05). Western blot analysis showed that the protein expressions of PI3K, p-AKT/AKT and p-mTOR/mTOR significantly decreased, the protein expressions of LC3 Ⅱ/Ⅰ, Atg5, and Beclin1 increased, and protein expression of P62 significantly decreased in lung tissues of the COPD rats compared with the normal rats (all P < 05). LC3B immunohistochemistry showed that the LC3B expression was higher in the COPD rats than that in the normal rats. ConclusionThe level of autophagy significantly increases in COPD rats with decreased expression of upstream proteins in autophagy signal pathway and increased expression of autophage proteins.
ObjectiveTo investigate the role of PI3K/AKT/mTOR signaling pathway in skeletal muscle atrophy in rats with chronic obstructive pulmonary diseases(COPD). MethodsPassive cigarette smoking was used to establish COPD model.The protein expression of PI3K, total mTOR, phosphorylated-mTOR, total GSK-3β, phosphorylated-GSK-3β, total 4E-BP1, phosphorylated-4E-BP1, total p70S6K1 and phosphorylated-p70S6K1 in extensor digitorum longus of rats were measured by Western blot. ResultsThe protein expression of PI3K was not significantly different between two groups(P > 0.05).Compared with the control group, the protein expression of total mTOR, phosphorylated-mTOR, total GSK-3β, and phosphorylated-GSK-3βincreased significantly in the COPD group(P < 0.05).The protein expression of total 4E-BP1 and total p70S6K1 were not significantly different between two groups(P > 0.05).While the protein expression of phosphorylated-4E-BP1 and phosphorylated-p70S6K1 significantly increased in the COPD group(P < 0.05). ConclusionThe protein expressions of PI3K/AKT/mTOR signaling pathway in extensor digitorum longus increased significantly in COPD rats, suggesting that the activity of PI3K/AKT/mTOR signaling pathway increased, which may be one of the compensatory mechanism of skeletal muscle atrophy in COPD.
ObjectiveTo describe the clinical,radiographic,and laboratory features of autoimmune pulmonary alveolar proteinosis (PAP) from a single center. MethodsConsecutive autoimmune PAP cases diagnosed in the Nanjing Drum Tower Hospital between January 2006 and December 2012 were recruited in the study. The clinical,radiographic and laboratory data of the PAP patients were analyzed to explore the clinical significance of serum GM-CSF autoantibody (GMAb) and serum cytokeratin (CYFRA21-1). ResultsThe median serum GMAb level of the 26 cases was 28.64 μg/mL (interquartile range,19.2-75.4 μg/mL),which were diagnosed as autoimmune PAP based on the serum GMAb levels of these patients all above the cut-off value of 2.39 μg/mL while the serum GMAb levels of 30 normal controls were 0.10(0.05-0.15)μg/mL and all below the cut-off value. 34.6% of all recruited 26 autoimmune PAP patients had identified occupational inhalational exposure. There was no significant correlation in the serum GMAb in autoimmune PAP patients with disease severity scores (DSS),lung function parameters,chest high resolution computed tomography (HRCT) scores,or PaO2 (P>0.05). There was significant correlation of DSS of autoimmune PAP patients with PaO2,FVC%pred,TLCO%pred,opacity extent score of chest HRCT,and opacity severity score of chest HRCT (P<0.05). The median serum level of CYFRA21-1 of the autoimmune PAP patients was 9.9(4.3-19.5)ng/mL,which was significant higher than that of the normal control group (P<0.05). However there was no significant correlation in the serum CYFRA21-1 in the autoimmune PAP patients with DSS,lung function parameters,and chest HRCT scores. 92.3% of the chest HRCT of 26 autoimmune PAP patients had crazy paving sign,while 100% of them had geographic sparing sign. ConclusionSerum GMAb and CYFRA21-1 may be important biomarkers for diagnosis of autoimmune PAP. The PAP with occupational inhalational exposure constitutes a high proportion of autoimmune PAP patients.
ObjectiveTo investigate the role of autophagy-lysosomal system in skeletal muscle atrophy in rats with chronic obstructive pulmonary disease (COPD). MethodsPassive cigarette smoking was used to establish COPD model. The mRNA and protein expression of FOXO transcription factor and autophagy-related genes Bnip3, Beclin1, p62, MAP-LC3Ⅱ/Ⅰ, Atg5 in extensor digitorum longus of rats were measured by real time PCR and Western blot. The changes of extensor digitorum longus tissue sections and lung tissue sections in the experimental group rats were observed under transmission electron microscopy. ResultsCompared with the control group, the mRNA expression of FOXO transcription factor and autophagy-related genes Bnip3, Beclin1, p62, Atg5 in extensor digitorum longus of the experimental group group rats was significantly increased (all P<0.05, as for Bnip3, the P value between two groups <0.01); The mRNA expression of MAP-LC3Ⅱ/Ⅰwas not significantly different between two groups (P>0.05). The protein expression of FOXO, Bnip3, Beclin1, p62, MAP-LC3Ⅱ/Ⅰ, Atg5 significantly increased in the COPD group (all P<0.05, as for Bnip3, MAP-LC3Ⅱ/Ⅰ, Beclin1, the P values between two groups <0.01). Compared with the control group, autolysosome in extensor digitorum longus tissue sections of the experimental group rats increased and lung tissue fibrosis and more inflammatory cells were observed in lung tissue sections of the experimental group rats under transmission electron microscopy. ConclusionThe mRNA and protein expressions of FOXO transcription factor and autophagy-related genes in extensor digitorum longus increase significantly in the experimental group rats, suggesting that the activity of autophagy-lysosomal system, which may be one mechanism of skeletal muscle atrophy in COPD.
Objective To observe the morphological changes of macular capillary in type 2 diabetic mellitus (DM) patients without clinical features of diabetic retinopathy (DR) by optical coherence tomography angiography (OCTA). Methods This is a prospective clinical case-control study. Forty-three eyes of 22 patients with DM without clinical features of DR (case group) and 40 control eyes of 20 age- and sex-matched healthy physical examination subjects (control group) were enrolled in this study. All subjects underwent OCTA examination with mode of retinal blood flow imaging, macular 3 mm×3 mm and 6 mm×6 mm area, signal strength ＞45. Foveal avascular zone (FAZ) area, foveal capillary density, parafovea capillary non-perfusion, and micro-aneurysm in shallow capillary vessel layer were evaluated. Results In case group, the mean FAZ area was (0.397±0.141) mm2 and the mean foveal capillary density was (44.6±0.62) %. In control group, the mean FAZ area was (0.253±0.112) mm2 and the mean foveal capillary density was (48.6±0.58) %. FAZ area of eyes in case group was larger than that in control group (t=1.017,P＜0.05). There was no difference of foveal capillary density between two groups (t=1.499,P＞0.05). The spider web-like FAZ and normal foveolar avascular zone were observed in eyes of control group. The parafovea capillary non-perfusion, abnormal foveolar avascular zone, micro-aneurysm and tortuosity of vessels were observed in eyes of case group. Parafovea capillary non-perfusion (χ2=4.542), micro-aneurysms (χ2=5.183) were seen more often in case group than control group (P＜0.05). Conclusion Type 2 DM patients have abnormal retinal vascular microcirculation before DR using OCTA, including larger FAZ area, parafovea capillary non-perfusion, abnormal foveolar avascular zone, micro-aneurysm and tortuosity of vessels.
ObjectiveTo investigate the clinical, radiographic characteristics and differential diagnosis of pulmonary Langerhans cell histiocytosis (PLCH) mimicking metastasis of cancer in radiography. MethodsClinical data of 2 patients with PLCH manifesting as metastatic cancer on HRCT and PET/CT were retrospectively analyzed. Patients reported as PLCH on WanFang Database, China Knowledge Resource Integrated Database and Pubmed were reviewed to screen misdiagnosis literature and further analyzed the clinical and radiographic characteristics. ResultsTwo cases both presented with cough and sputum. 18F-FDG PET/CT showed increased 18F-FDG up-take in both nodules in the lungs. One patient presented with multiple nodules, diffuse multiple cystic changes in lungs and osteoclasia in the right 4th rib on HRCT who was diagnosed by a video-assisted thoracoscopic biopsy of rib biopsy. The other patient presented with diffuse multiple nodules on HRCT who was diagnosed by a video-assisted thoracoscopic biopsy of lung biopsy. The pathological characteristics of both biopsy specimen demonstrated infiltration by Langerhans cells (LC) and eosinophils. The LC were positive for CD1a. Literature review found seven PLCH cases who were misdignosed as depression, eosinophilic pneumonia, interstitial lung disease involvement of autoimmune disorders and malignant tumor. ConclusionWhen clinician faced with a patient suspected as metastatic cancer by HRCT and PET/CT, it is reasonable to consider PLCH as a differential diagnosis and obtain the pathological information as soon as possible so that better prognosis can be achieved through early intervention.