Objective: To investigate the efficacy of bronchoalveolar lavage (BAL) and its influence factors in the treatment of Mycoplasma pneumoniae pneumonia (MPP) with atelectasis. Methods: A retrospective case control study was performed on hospitalized MPP patients with atelectasis and received BAL in the Department of Pulmonology, Children's Hospital Zhejiang University School of Medicine from January 1, 2015 to July 31, 2017. Fever relieved in 48 hours and chest imaging improved in one week after BAL were considered effective. Clinical data, including age, sex, blood routine tests, lactate dehydrogenase (LDH), cytokines, complications, fever duration before BAL, course of disease before BAL, sputum plug, atelectasis area and its CT values of atelectasis site were collected. Student's t test, Mann-Whitney U test, or chi square test were used. Results: (1) A total of 163 patients were enrolled, including 69 boys and 94 girls, with the ratio of 1∶1.36. Their ages ranged from 6 months to 12.6 years. (2) On the day of bronchoscope, 113 patients still had fever. They were divided into effective group (n=66) and ineffective group (n=47) according to whether fever was relieved in 48 hours after BAL. The effective group were found to have less sputum plug compared with the ineffective group (33% (22/66) vs. 57% (27/47), χ²=6.499, P=0.011). The other factors such as sex, age, fever duration before BAL, course of disease before BAL, C reactive protein (CRP), LDH, IL-2, IL-4, IL-6, IL-10, TNF, IFN-γ, atelectasis area and CT value showed no significant difference between the two groups (all P>0.05). (3)A total of 122 cases had chest imaging after BAL. According to chest imaging improvement, they were divided into effective group (n=81) and ineffective group (n=41). The effective group showed lower CT value ((58±9) vs. (63±8) HU, t=-2.436, P=0.017), IL-6 and IL-10 (M(Q₁, Q₃)) (21.0 (1.9, 48.4) vs. 36.4(21.8, 93.6), 4.9 (3.7, 9.6) vs. 7.7 (4.4, 12.0) ng/L, Z=-2.387,-2.009, P=0.017, 0.045). Sex, age, fever duration before BAL, course of disease before BAL, CRP, LDH, IL-2, IL-4, TNF, IFN-γ, atelectasis area showed no significant differences between the two groups (all P>0.05). (4) Patients were divided into sputum plug group (57 cases) and non sputum plug group (106 cases) according to bronchoscopic findings. The sputum plug group showed higher LDH, CRP, IL-6, IFN-γ, incidence of pleural effusion and extrapulmonary complications (585(433, 833) vs. 369 (312, 588) U/L, 42 (19, 103) vs. 25 (12, 45) mg/L, 38.8 (22.1, 71.3) vs. 20.7 (9.2, 48.3) ng/L, 33.1 (13.5, 89.3) vs. 12.7 (6.5, 33.6) ng/L, 73.7% (42/57) vs. 52.8% (56/106), 40.4% (23/57) vs. 17.0% (18/106)), with statistically significant differences (Z=-4.865,-3.435,-3.098,-3.704, χ²= 0.010, 0.001, all P<0.01) . Additionally, fewer patients showed fever relief within 48 hours after BAL in the cases with sputum plug cases compared those without sputum plug (44.9% (22/49) vs. 68.8% (44/64), χ²= 0.011, P=0.009). Fewer patients showed chest imaging improvement within one week after BAL in the cases with sputum plug compared with those without sputum plug, but did not show significant difference (56.5% (26/46) vs. 72.4% (55/76), χ²=0.073, P=0.056). Conclusions BAL has some therapeutic effect on fever or atelectasis in MPP children complicated with atelectasis. Chest imaging improvement or fever relief may be hampered by sputum plug, increased IL-6 or IL-10.

ObjectiveTo explore the mechanism of Qigesan （QGS） in intervening in the migration and invasion of esophageal carcinoma TE-1 cells. MethodMicroarray technology was used to screen differentially expressed genes （DEGs） in the normal group and the QGS group， and the ontological functions and signaling pathways of DEGs were analyzed. The thiazolyl tetrazolium （MTT） assay was used to detect the effect of QGS on the viability of TE-1 cells. In the subsequent experiments for verification， a blank group， a transforming growth factor-β₁ （TGF-β₁） group， a TGF-β₁ + QGS group， and a TGF-β₁ + SB431542 group were set up. The cell morphology in each experimental group was observed by microscopy. The migration and invasion abilities of cells were detected by wound healing assay， and the mRNA expression levels of E-Cadherin， vimentin， Smad2， and Smad7 were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression of E-Cadherin， vimentin， p-Smad2/3， Smad2/3， and Smad7 was detected by Western blot. ResultThere were 1 487 DEGs between the QGS group and the blank group， including 1 080 down-regulated ones （accounting for 72.63%） and 407 up-regulated ones. The down-regulated genes were mainly involved in biological processes such as cytoskeletal protein binding， ATP binding， adenylate nucleotide binding， and adenylate ribonucleotide binding， and the involved Kyoto Encyclopedia of Genes and Genomes （KEGG） pathways included TGF-β signaling pathway， cell cycle， extracellular matrix-receptor interaction protein， tumor pathways， and oocyte meiosis. The up-regulated genes were mainly involved in RNA binding， DNA binding， transcriptional regulator activity， transcriptional activator activity， and nucleotide binding， and the KEGG pathways involved mainly included mitogen-activated protein kinase （MAPK） signaling pathway， bladder cancer， renal cell carcinoma， cancer pathways， and p53 signaling pathway. Compared with the blank group， the inhibition rate of cell viability of TE-1 cells increased after QGS （20， 30， 40， 60， 80 mg·L^-1） intervention for 12， 24， 36， 48， 60 h （P<0.05）， and the inhibition rate was time- and dose-dependent. Compared with the blank group， the TGF-β₁ group showed lengthened cells with fibroblast phenotype. Compared with the TGF-β₁ group， the TGF-β₁ + QGS group showed shortened cells with normal morphology and epithelial phenotype. The cell morphology in the TGF-β₁ + SB431542 group was similar to that of the TGF-β₁ + QGS group. Compared with the blank group， the TGF-β₁ group showed potentiated ability of cell migration and invasion （P<0.05）. Compared with the TGF-β₁ group， the TGF-β₁ + QGS group and the TGF-β₁ + SB431542 group showed inhibited and weakened migration and invasion abilities of cells （P<0.05）. However， there was no significant difference in migration and invasion abilities between the TGF-β₁ + QGS group and the TGF-β₁ + SB431542 group. The mRNA expression levels of vimentin and Smad2 in the TGF-β₁ group were higher （P<0.05）， and the mRNA expression levels of E-Cadherin and Smad7 were lower （P<0.05） than those in the blank group. Compared with the TGF-β₁ group， the TGF-β₁ + QGS group and the TGF-β₁+ SB431542 group exhibited decreased expression levels of vimentin and Smad2 mRNA （P<0.05）， and elevated expression levels of E-Cadherin and Smad7 mRNA （P<0.05）. Compared with the blank group， the TGF-β₁ group showed up-regulated protein expression levels of vimentin， p-Smad2/3， and Smad2/3 （P<0.05）， and reduced protein expression levels of E-Cadherin and Smad7 （P<0.05）. Compared with the TGF-β₁ group， the TGF-β₁ + QGS group and the TGF-β₁ + SB431542 group displayed decreased protein expression levels of vimentin， p-Smad2/3， and Smad2/3 （P<0.05）， and increased protein expression levels of E-Cadherin and Smad7 （P<0.05）. ConclusionThe ethyl acetate extract of QGS inhibits the epithelial-mesenchymal transition （EMT） of TE-1 cells through the TGF-β₁ pathway to reduce the migration and invasion of TE-1 cells.

ObjectiveTo investigate the mechanism of icariin in ameliorating efferocytosis dysfunction and inflammatory response of alveolar macrophages induced by cigarette smoke extract via the peroxisome proliferator-activated receptor gamma （PPARγ） signaling pathway. MethodThe untreated rat alveolar macrophages （NR8383） were taken as the blank group. The NR8383 cells treated with 10% cigarette smoke extract were divided into model， low-， medium-， and high-dose （10， 20， 40 μmol·L^-1） icariin， PPARγ inhibitor， and PPARγ inhibitor + low-， medium-， and high-dose icariin groups. Alamar blue colorimetry was employed to examine the proliferation and toxicity of icariin on NR8383 cells. The efferocytosis rate of NR8383 cells was detected by flow cytometry. Enzyme-linked immunosorbent assay was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， transforming growth factor-β₁ （TGF-β₁）， and milk fat globule-epidermal growth factor 8 （MFG-E8）. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were employed to determine the protein and mRNA levels， respectively， of PPARγ， CD36， and RAS-related C3 botulinum toxin substrate 1 （Rac1）. ResultThe efferocytosis dysfunction model of NR8383 was established with the cigarette smoke extract. Compared with the blank control group， the model group showed decreased efferocytosis rate （P<0.05）， elevated TNF-α level （P<0.05）， lowered TGF-β₁ and MFG-E8 levels （P<0.01）， and down-regulated mRNA and protein levels of PPARγ， CD36， and Rac1 （P<0.05， P<0.01）. Compared with the model group， the treatment with icariin increased the efferocytosis rate （P<0.05， P<0.01）， lowered the TNF-α level （P<0.01）， elevated TGF-β₁ and MFG-E8 levels （P<0.05）， and up-regulated the protein and mRNA levels of PPARγ， CD36， and Rac1 （P<0.05， P<0.01）. Compared with icariin alone， PPARγ inhibitor + icariin decreased the efferocytosis rate （P<0.05） and down-regulated the protein and mRNA levels of PPARγ （P<0.05， P<0.01）. In addition， PPARγ inhibitor + low-dose icariin down-regulated the protein level of CD36 （P<0.01） and PPARγ inhibitor + low-/medium-dose icariin up-regulated the protein level of Rac1 （P<0.05）. ConclusionIcariin ameliorates the cigarette smoke extract-induced efferocytosis dysfunction of alveolar macrophage by regulating the PPARγ signaling pathway and cytoskeletal structure rearrangement.