AIM: To explore the effects of p38 mitogen-activated protein kinases (MAPK) and phosphoinositide 3-kinases (PI3K)/Akt on interleukin (IL)-6, the endothelin (ET)-1-mediated process of airway fibroblast activation induced by injured human bronchial epithelial cells (HBE). METHODS: Human primary cultured airway fibroblasts were co-cultured with HBE pre-treated with or without poly-L-arginine (PLA). The procedure was also performed in the presence or absence of p38 MAPK selective inhibitor SB203580, PI3K selective inhibitor LY294002 or ETA receptor blocker BQ123, respectively. Immunostaining, Western blotting or ELISA were used for detecting α-smooth muscle actin (α-SMA) expression, the activities of p38 MAPK and Akt in fibroblasts or IL-6 levels in supernatants of fibroblasts. In addition, fibroblasts were mixed with soluble collagen and cultured with HBE treated as the same mentioned above, the gel contraction was measured by serial area measurements. RESULTS: ET-1 and IL-6 levels [(13.69±1.36) ng/L, (56.7±10.7) ng/L] in the supernatants of fibroblasts cultured with injured HBE were significantly higher than those in the supernatants of fibroblasts cultured with HBE [(3.79±0.64) ng/L, (15.5±3.2) ng/L]. BQ123, SB203580 or LY294002 decreased IL-6 levels [(27.2±3.1) ng/L, (31.5±3.6) ng/L, (41.3±3.2) ng/L] differently in the supernatants of fibroblasts induced by injured HBE. Activation of p38 MAPK preceded Akt in fibroblasts cultured with injured HBE. BQ123 reduced the phosphorylation levels of p38 MAPK and Akt. SB203580 concentration-dependently attenuated Akt phosphorylation, while LY294002 had little effect on p38 MAPK phosphorylation. Fibroblasts expressed more α-SMA after cultured with injured HBE and showed significant increase in the gel contraction compared to fibroblasts cultured with HBE [percentage of gel contraction: (61.2±2.7)% vs (15.4±7.3)%], all these effects were diminished or inhibited by BQ123, SB203580 or LY294002. Furthermore, the effects of BQ123 and SB203580 on decreased gel contraction were stronger than the effect of LY294002. CONCLUSION: ET-1 exerts a key role in the airway fibroblasts activation induced by injured HBE through activating p38 MAPK, PI3K/Akt signaling and promoting IL-6 expression.

AIM:To investigate the regulatory role of hypoxia mimic reagent cobalt chloride ( CoCl2 ) on cave-olin-1 (Cav-1) generation and the influence of Cav-1 on the abilities of migration and invasion of human lung adenocarcino-ma A549 cells.METHODS:The concentrations of Cav-1 and hypoxia-inducible factor ( HIF)-1αin pleural effusion of the patients with lung cancer ( MPE) or tuberculous pleurisy ( TBPE) were detected, and the correlation was also compared. A549 cells were treated with CoCl2 at different concentrations and time in the presence or absence of HIF-1αinhibitor YC-1.The concentrations of Cav-1 and HIF-1αin the cell supernatants were measured by ELISA.The effects of Cav-1 induced by CoCl2 on the migration and invasion of A549 cells were determined by scratch test and Transwell invasion trial, respec-tively.RESULTS:The levels of Cav-1 and HIF-1αin MPE were significantly higher than those in TBPE.There was a highly positive correlation between Cav-1 and HIF-1αlevels in the pleural effusion.CoCl2 induced the generation of Cav-1 and HIF-1αin A549 cells in a concentration-and time-dependent manner, the peak occurred at 200 μmol/L or 24 h, while the concentration over 200 μmol/L or after treated over 24 h, a concentration-or time-dependent inhibition was ob-served.HIF-1αinhibitor YC-1 concentration-dependently inhibited the generation of HIF-1αand Cav-1 induced by CoCl2 in A549 cells.CoCl2 enhanced A549 cells migration and invasion, with 200 μmol/L played the strongest role, which were down-regulated significantly in the presence of YC-1.CONCLUSION:The alteration of hypoxia-induced Cav-1 generation might be involved in the migration and invasion of A549 cells.A possible role for HIF-1αis indicated in Cav-1 generation.

Objective To introduce the application of the network training on clinicians'knowledge of malaria diagnosis and treatment in Yunnan Province,and evaluate its effect. Methods Through the platform Yiboshi(www.yiboshi.com),the medi-cal and health personnel at the units of provincial,prefectural,county levels and 25 townships of 25 border counties were trained on the knowledge of malaria diagnosis,treatment,prevention and control,and the effects were evaluated by examina-tions,questionnaires and interviews. Results Totally 7152 participants were trained,the average participation,completion and pass rates of the training were 95.26%,98.55% and 97.30%,respectively. The trainees mainly learned malaria control knowledge from 3 aspects,namely policy of malaria elimination,malaria epidemiology,malaria diagnosis and treatment. The questionnaires showed that 95.94%of the participants considered that their theoretical and technical levels improved,97.30%were interested in the training content,93.24% recognized the arrangement of the training time was reasonable,and 91.89%were satisfied with the service of the platform. Conclusions The network training on knowledge of malaria diagnosis and treat-ment in Yunnan Province has achieved good effect. The network training meets the need of training a large number of clinicians in the malaria elimination and post-elimination stage.

Objective To explore the dynamic change and clinical signiticance of plasma D-damer level in patients with cerebral hemorrhage and acute craniocerebral injury.Methods 50 patients with cerebral hemorrhage and 40 patients with acute craniocerebral injury were selected,The enzyme-linked immunosorbent assay (ELISA) was used to measure plasma D-dimer level in two groups of patients after onset,and the results were compared with 40 healthy controls.Results The levels of plasma D-dimer in the patients with cerebral hemorrhage were 1.59mg/L,2.10mg/L,1.03 mg/L,0.82mg/L at 3 h,6h,12h,2d after onset,which in the patients with acute craniocerebral injury were 1.61mg/L,2.02mg/L,1.01mg/L and 0.67mg/L,respectively.And the plasma D-dimer levels were 0.50mg/L,0.49mg/L,0.47mg/L,0.48mg/L in the control group at 3h,6h,12h and 2d after onset.The levels of plasma D-dimer in the patients with acute craniocerebral injury were significantly higher than those in the control group,and the differences were statistically significant (t =9.35,12.17,4.03,3.05,all P ＜ O.05).At 7d after onset,the D-dimer levels in the cerebral hemorrhage group and acute craniocerebral injury group were 0.53mg/L,0.55mg/L,respectively,which of the control group was 0.47mg/L,there was no statistically significant difference among the three groups(P ＞ 0.05).Conclusion Cerebral hemorrhage patients and acute craniocerebral injury patients have high coagulation and fibrinolytic activity in brief increase trend,dynamic observation of plasma D-dimer level in patients with cerebral hemorrhage and acute craniocerebral injury is helpful to determine courses,condition and evaluate prognosis.

Objective To compare the clinical characteristics of chronic bronchitis ( CB),emphysema (EM ), asthma - chronic obstructive pulmonary disease overlapping syndrome ( ACOS ) with frequent exacerbations ( FE ) or infrequent exacerbations ( iFE ) and induced sputum inflammatory cells and the heterogeneity of the transmitter. Methods Ninety-one cases of chronic obstructive pulmonary disease( COPD) with acute exacerbation were divided into CB,EM or ACOS phenotype,among which 44 were frequent,and 47 were non frequent. The clinical data,induced sputum inflammatory cells,interferon-γ(IFN-γ),tumor necrosis factor-α ( TNF-α ), interleukin ( IL )-4, IL-13 were analyzed. Results The FEV1% was ( 47 ± 13. 1 )%, significantly lower than that of non frequent episodes (( 56. 2 ± 10. 2)%),and the difference was statistically significant(P＝0.049).The FEV1/FVC% was (54.3±9.3)%,significantly lower than that of non frequent episodes (60. 1±7. 3)%,and there was a significant difference between them ( P＝0. 001) . The proportion of patients with GOLD III and IV,the percentage of neutrophils in induced sputum,tumor necrosis factor -α(TNF-α) and interferon-γin the patients with frequent episodes were significantly higher than those with non frequent episodes (P＜0. 05). Among them,FEV1/FVC% and TNF-αwere independent risk factors for COPD patients (P＝0. 032, 0. 021) . The FEV1% of patients with CB phenotypic frequent episodes were ( 47. 9 ± 14. 9 )%, significantly lower than that of non frequent episodes ((57. 2±10. 9)%)(P＝0. 000),and FEV1/FVC% was (53. 4± 9. 5)% in patients with CB frequent episodes,significantly lower than that of non frequent episodes ((60. 3±6. 9)%),and the difference was statistically significant (P＝0. 022),while the level of N%,TNF-α in induced sputum were significantly higher in CB phenotype subjects with FE than those in subjects with iFE(P＜0. 01). Patients with frequent episodes of emphysema had longer duration of disease (P＜0. 05),lower FEV1%and FEV1/FVC%(P＜0. 05),the proportion of GOLD III patients and the induced sputum TNF-αwere higher, but there was no significant difference in the number and proportion of phlegm inflammatory cells,interferon-γ, interleukin 4 and interleukin 3. The level of GOLD III and the IL-13 level of induced sputum in patients with frequent ACOS phenotype were significantly higher than those in patients with non frequent episodes (P＜0. 05) . Conclusion The lung function,the severity of the disease,the course of the disease,and the percentage of sputum neutrophils,tumor necrosis factor-α,or interleukin 13 are helpful in diagnosing patients with high risk of frequent episodes.

OBJECTIVETo investigate the characteristics of multifocal electroretinograms (mf-ERG) of different phases in diabetic retinopathy (DR) and its clinical significance.

METHODSMultifocal electroretinograms in patients with DR (I - II stage) were tested with VERIS IV system.

RESULTSIn I - II stage, the absolute values of N1, P1 and N2 response densities, and the N1-P1 and P1-N2 response densities were attenuated is a field of about 45 degrees in diameter.

CONCLUSIONAs a new objective and quantitative examination for spatial visual function, multifocal electroretinograms may be valuable in the diagnosis of diabetic retinopathy.

Aged ; Diabetic Retinopathy ; pathology ; physiopathology ; Electroretinography ; methods ; Female ; Humans ; Male ; Middle Aged ; Retina ; pathology ; physiopathology ; Time Factors

To investigate the effects of airway epithelial cells on macrophages chemotaxis and inflammatory cytokine expression under hypoxic conditions.
 Methods: Human bronchial epithelial cells (HBE) treated with different concentrations (0, 100, 200, 400, 800 μmol/L) of CoCl2 or transfected with HIF-1α siRNA were co-cultured with THP-1-derived M1 macrophages or M2 macrophages. The chemotactic effects on macrophages were analyzed by Transwell assay. The levels of TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants of macrophages were detected by ELISA, and HIF-1α or Cav-1 mRNA expression in HBE or macrophages was detected by RT-qPCR.
 Results: HBE cells promoted macrophages chemotaxis in a time- and concentration-dependent manner. Compared to un-transfected group, the chemotactic ability of HBE transfected with HIF-1α siRNA was significantly weakened (P<0.01). Under the same culture conditions, the chemotaxis of M2 macrophages was greater than that in THP1-derived M1 macrophages. The concentrations of TNF-α, IFN-γ, IL-4, IL-13 and IL-10 in the supernatants of macrophages were increased in a time-and concentration-dependent manner. The concentrations of TNF-α and IFN-γ were increased further after co-culturing for 8 and 12 h; while IL-4, IL-13 and IL-10 concentrations were increased further during 24 h of co-culture. The levels of cytokines in the supernatants of macrophages co-cultured with HBE and transfected with HIF-1α siRNA were significantly lower than those in un-transfected cells (P<0.05 or P<0.01). The reduction of TNF-α or IFN-γ was more obvious. The expression of HIF-1α or Cav-1 mRNA in HBE or macrophages was increased in a concentration-dependent manner after 8 or 12 h co-culture, which was significantly reduced when HBE was transfected with HIF-1α siRNA.
 Conclusion: Airway epithelial cells can enhance macrophages chemotaxis and pro-inflammatory cytokines expressions under hypoxic condition. HIF-1α and Cav-1 may be the important mediators in these processes.
Cell Hypoxia ; Chemotaxis ; Cytokines ; Epithelial Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Macrophages

Fungal keratitis is a serious blinding eye disease. The development of fungal infections depends primarily on the interaction of fungal virulence with host immune defense factors. The cornea is considered an immune-privileged organ, and resident macrophages are the main immune cells that respond to the heterogeneity exhibited by the microenvironment with their polarization. In the early stage of infection, macrophages polarize towards M1, which promotes inflammation and facilitates fungal clearance but produces a cellular storm that exacerbates immune damage; in the late stage of infection, macrophages polarize towards M2, which suppresses the inflammatory response and facilitates tissue repair, but may be immunosuppressed or even immune escape to the detriment of pathogen clearance. The balance between pro-inflammatory and anti-inflammatory responses is key to maintaining the functional integrity of the cornea. Current antifungal drug therapy is limited, so it is particularly important to find a therapeutic target for the inflammatory response triggered by the immune response in addition to antifungal therapy. In this review, the functional and phenotypic characterization of macrophage subsets associated with fungal keratitis was reviewed, more in-depth research is needed to explore the specific mechanisms by which macrophage polarization and their impact on fungal keratitis. Targeted regulation of macrophage differentiation based on their phenotype and function could be an effective approach to treat and manage fungal keratitis in the future.