Menthol enhances interleukin-13-induced synthesis and secretion of mucin 5AC in human bronchial epithelial cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of interleukin (IL) -13 combined with cold stimulation on synthesis and secretion of mucin (MUC) 5AC in human bronchial epithelial cell line 16HBE and explore the role of transient receptor potential 8 (TRPM8) and anti-apoptotic factor B-cell lymphoblast-2 (Bcl-2) in this process.@*METHODS@#16HBE cells were stimulated with 10 ng/mL IL-13, 1 mmol/L menthol, or both (1 mmol/L menthol was added after 6 days of IL-13 stimulation), and the changes in the expression of MUC5AC, intracellular Ca@*RESULTS@#The mRNA and protein expressions of MUC5AC increased significantly in 16HBE cells following stimulation with IL-13, menthol, and both (@*CONCLUSIONS@#Menthol combined with IL-13 produces a synergistic effect to promote the synthesis and secretion of MUC5AC in 16HBE cells possibly by activating TRPM8 receptor to upregulate the expression of Bcl-2.

Occupational burnout of vaccination personnels in Haining / 预防医学

Objective @#To understand the current situation and influencing factors of occupational burnout among vaccination personnels in Haining.@*Methods @#The vaccination staffs of all vaccination clinics in Haining were investigated by the general questionnaire and Maslach Burnout Inventory. Logistic regression model was used to analyze the influencing factors for job burnout of vaccination personnels. @*Results @#A total of 160 questionnaires were distributed and 158 valid questionnaires were collected. The effective rate of questionnaires was 98.75%. A total of 91 vaccination staffs suffered from occupational burnout,accounting for 57.59%. Among them,the median（inter-quartile range）of the scores of emotional exhaustion,depersonalization and personal achievement were 13.00（14.00）,4.00（6.00）and 26.50（17.00）,respectively,which were all lower than the normalized scores（22.19,7.12 and 36.53,P<0.05）. The results of logistic regression analysis showed that having confidence in vaccination was a protective factor for emotional failure（OR=0.175,95%CI：0.058-0.523）and low sense of achievement（OR=0.272,95%CI：0.079-0.937）in vaccination personnels;having experience in adverse event following immunization（AEFI）was a risk factor for depersonalization（OR=3.125,95%CI：1.472-6.633）and occupational burnout（OR= 2.391,95%CI：1.189-4.807）in vaccination personnels. @*Conclusion @#A certain proportion of vaccination staffs in Haining suffered from occupational burnout. The experience of AEFI was a risk factor for their occupational burnout.

Clinical and inflammatory phenotypic features of asthmatic patients sensitive to cold stimulation / 南方医科大学学报

OBJECTIVE@#To explore the clinical symptoms, lung function and airway inflammation phenotype characteristics of asthmatic patients who are sensitive to cold stimulation.@*METHODS@#Eighty patients with newly diagnosed bronchial asthma or with mild to moderate acute exacerbation of previously diagnosed bronchial asthma but without regular treatment were selected. According to whether cold air stimulation could induce respiratory symptoms such as cough and wheeze, the patients were divided into cold-insensitive group (45 cases) and cold-sensitive group (35 cases). All the patients were treated with inhaled corticosteroid (ICS), long-acting β2 receptor agonist (LABA; salmeterol xinafoate and fluticasone propionate powder for inhalation, 50 μg/250 μg, twice daily) and montelukast sodium tablets (10 mg, once daily); short-acting β2 receptor agonist (SABA) and/or systemic glucocorticoid (prednisone acetate tablets, 10 mg, once daily; or injection of methylprednisolone sodium succinate, 40 mg) were given if necessary. Asthma Control Test (ACT) score before treatment and at 3 months of treatment was used to assess the clinical symptoms such as cough and wheeze; spirometry was performed to determine lung function impairment and recovery. Blood and induced sputum cell counts were examined to determine the characteristics of airway inflammation.@*RESULTS@#The two groups were comparable for age, gender, BMI, proportion of smokers and allergic rhinitis before treatment. The cold-sensitive patients experienced significantly more frequent acute exacerbations than the cold-insensitive patient within 1 year before the visit ( < 0.05), but the use of SABA and glucocorticoid for symptom control during the treatment did not differ significantly between the two groups ( > 0.05). The ACT scores of the cold-sensitive group were significantly lower than those of the cold-insensitive group both before and after the treatment ( < 0.01). Compared with the cold-insensitive patients, the cold-sensitive patients had more obvious impairment of FEV1/FVC% and FEV1%pred before treatment ( < 0.01), and also showed poorer recovery after treatment ( < 0.05). The percentages of eosinophils in blood and induced sputum samples did not differ significantly between the two groups either before and after the treatment, but the percentage of neutrophils was significantly higher in the cold-sensitive group ( < 0.01). In the induced sputum samples collected before treatment, the cell populations consisted mainly of eosinophilic subtype (60%) and neutrophilic subtype (20%) in the cold-insensitive group; in the cold-sensitive patients, the sputum neutrophilic subtype cells increased significantly to 42.86% (=0.03) and the eosinophilic subtype cells were lowered to 31.43% (=0.01).@*CONCLUSIONS@#The cold-sensitive asthmatic patients experience frequent recurrent and/or aggravated symptoms and have obvious lung function impairment. Different from that in patients with classic asthma, the airway inflammatory phenotype in these patients is characterized by the domination by neutrophilic subtype.

High pressure level in human airwayenhances the expression of 3 cytokines related to airway remodeling / 基础医学与临床

Objective To explore the relationship between different airway pressure and expression of related cytokines to airway remodeling.Methods Fourty-two chronic obstructive pulmonary disease cases (COPD group) and thirty-three control cases were collected.The above cases underwent mechanical ventilation in the period of general anesthesia.According to different levels of peak inspiratory pressure(PIP), above two groups, randomly and respectively, were divided into high PIP (HPIP, 24 cmH2O) group, moderate PIP (MPIP, 22 cmH2O) group, low PIP (LPIP, 20 cmH2O) group.All positive end expiratory pressure (PEEP) was set to 5 cmH2O.Then the collection of BALF was performed before and 3 hours after applying ventilator.The related factors to airway remodeling, such as fibroblast growth factor 2 (FGF-2), transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-9 (MMP-9), were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot.Results 1)Beforemechanical ventilation, the levels of FGF-2, TGF-β1 and MMP-9 in COPD group were higher than control group (P<0.01).2)3 hours after mechanical ventilation, we saw significant upregulated expression of FGF-2, TGF-β1, MMP-9 in HPIP subgroup in control group (P<0.05) and the 3 factors levels of COPD group were all increased (P<0.05).Moreover, HPIP subgroup was significantly higher than MPIP and LPIP subgroup in COPD group (P<0.05).3)The expression of FGF-2, TGF-β1, MMP-9 in BALF had a positive correlation with the airway pressure levels in COPD group(P<0.01).Conclusions Under mechanical ventilation, sustained high airway pressure may enhance the expression of FGF-2, TGF-β1 and MMP-9 which may result in airway remodeling by mechanosensitive cation channel in bronchial epithelial cells, especially in COPD patients.

Accurate Detemination of Isotopic Abundance of Intracellular Metabolites of Saccharopolysporaerythraea Based on Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry / 分析化学

A method for measuring 13C isotopic abundance of intracellular metabolites of Saccharopolysporaerythraea by ultra-high performance liquid chromatography (UPLC)-triple quadrupole mass spectrometry was established.First, the chromatographic conditions of UPLC were optimized, and then the MS conditions such as unique tube lens voltage, collision energy, and ion pair were optimized.On the bases of length of the parent and daughter ions carbon chains and whether the daughter ions contain 13C atoms, the one-to-one method, one-to-many method and SIM method were established for measuring 13C isotopic abundance.Then these methods were used to measure naturally labeled intracellular metabolite standards and 13C labeled samples, and according to the gap between the experimental value and the theoretical value, the best method was established for each metabolite of different characteristics.The results showed that one-to-one method was most effective for measuring the metabolites of daughter ions not containing 13C atoms represented by sugar phosphates, one-to-many method was the best for measuring the metabolites of both parent and daughter ions containing 13C short carbon chains represented by carboxylic acids, SIM method could play a role in measuring the metabolites of both parent and daughter ions containing 13C long carbon chains represented by coenzyme A.This method had a good measurement precision and could be applied to the measurement of Saccharopolysporaerythraea intracellular metabolites, which contributed to the consequent study of metabolic mechanism and the efficient expression of erythromycin.

Modulation of TLR-4/MyD88 signaling cascade by miR-21 is involved in airway immunologic dysfunction induced by cold air exposure / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of miR-21 in airway immunologic dysfunction induced by cold air irritation.</p><p><b>METHODS</b>Immortalized human airway epithelial cell lines BEAS-2B and 16HBE cells were cultured in air-liquid phases. The differential expressions of endogenous miR-21, miR-164, and miR-155 in the cells induced by cold air exposure for different time were detected by real-time PCR. The reporter plasmid containing wild-type or mutated 3'UTR of TLR-4 were constructed and co-transfected into BEAS-2B cells or 16HBE cells together with miR-21 mimic, miR-21 mimic control, miR-21 inhibitor, or miR-21 inhibitor control. Following the transfection, dual luciferase reporter assay was performed to verify the action of miR-21 on TLR-4. miR-21 mimic, miR-21 mimic control, miR-21 inhibitor, and miR-21 inhibitor control were transfected via lipofectamine 2000 in BEAS-2B or 16HBE cells that were subsequently exposed to a temperature at 37 degrees celsius; or cold irritation (30 degrees celsius;), and the protein levels of TLR-4/MyD88 were detected by Western blotting.</p><p><b>RESULTS</b>Cold irritation caused a time- dependent up-regulation of miR-21 in both BEAS-2B and 16HBE cells (P<0.05) without obviously affecting the expressions of miR-164 and miR-155. Dual luciferase reporter assay demonstrated a direct combination of miR-21 and its target protein TLR-4. The synthesis levels of TLR-4/MyD88 protein were decreased in miR-21 mimic group even at a routine culture temperature (P<0.05), as also seen in cells with cold irritation (P<0.05). Treatment with the miR-21 inhibitor partially attenuated cold irritation-induced down-regulation of TLR-4/MyD88 protein (P<0.05).</p><p><b>CONCLUSION</b>Cold air irritation-induced airway immunologic dysfunction is probably associated with TLR-4/MyD88 down-regulation by an increased endogenic miR-21.</p>

Analysis on correlation between expression level of pressure-sensitive channel protein and airway remodeling in COPD patients / 吉林大学学报(医学版)

Objective:To explore the expression of pressure-sensitive transient receptor potential channel (TRPC1)in bronchial epithelium cells of the patients with chronic obstructive pulmonary disease (COPD),and to explain the molecular mechanism about the participation of TRPC1 in airway remodeling.Methods:Sixty-four patients who needed fiberoptic bronchoscope for diagnosis were selected. All the patients were used as non-operation group and divided into COPD group (40 cases of mild to moderate grade)and control group (24 cases without chronic airway inflammatory disease)according to Respiratory Disease Guideline. The level of the forced expiatory volume/forced vital capacity (FEV1/FVC)in the first second and forced expiatory volume in the first second of the expected value (FEV1%pred)were detected by lung function test.The expression levels of TRPC1, matrix metalloproteinase (MMP-9)and transforming growth factor-β1 (TGF-β1 )in bronchoalveolar liquid (BALF) were measured by ELISA. The relevance among expression level of TRPC1 and levels of FEV1/FVC, FEV1%pred,MMP-9,TGF-β1 was analyzed.Seventeen lung samples from the patients underwent pulmonary lobectomy were selected.All the patients were used as operation group and divided into COPD group (8 cases of mild to moderate grade ) and control group (9 cases without chronic airway inflammatory disease ). Then indictator relevance to airway remodeling,such as thickness-diameter ratio (TDR%),percentage of wall thickness (WA%) were measured.The immunohistochemistry experiment was used to detect the distribution characteristics of TRPC1 protein in human bronchial epithelial cells. The Imagepro-plus 6.0 software was used to analyze the expression difference of TRPC1 protein between COPD and control groups. Western blotting method was used to detect the expression level of TRPC1 . The correlation between the expression level of TRPC1 and TDR%, WA% was analyzed.Results:The lung function detection results showed that in non-operation group,the levels of FEV1%pred and FEV1/FVC in COPD group were decreased compared with control group (P<0.05).The expression levels of TRPC1,MMP-9 and TGF-β1 in COPD group were increased compared with control group (P<0.05). The expression level of TRPC1 was negatively correlated with FEV1%pred (r= -0.34,P=0.002)and FEV1/FVC (r= -0.38,P=0.004).The expression level of TRPC1 was positively correlated with MMP-9 (r=0.36, P=0.004)and TGF-β1 (r=0.61,P=0.002).The immunohistochemistry results in operation group displayed that the TRPC1 protein was mainly distributed in the epithelial columnar cell nucleus and the cell membrane near lumen within the bronchial epithelium.The ratio of integrated optical density (IOD)to area within the bronchial epithelium in COPD group was significantly higher than that in control group (P<0.05).The expression level of TRPC1 protein detected by Western blotting method in COPD group was higher than that in control group (P<0.05).The expression level of TRPC1 was positively correlated with TDR% (r=0.59,P=0.002)and WA%(r=0.60,P=0.002).Conclusion:The TRPC1 channel in human bronchial epithelial cells of COPD patients is actived and up-regulated.The expression level of TRPC1 channel is positively correlated with impared extent of lung function and the expression levels of cytokines relevant to airway remodeling. All above imply that pressure-sensitive TRPC1 channel could promote the development of airway remodeling.

Accurate Determination of 13 C Isotopic Abundances of Free Intracellular Amino acids with Low Concentration by GC-MS-Selective Ion Monitoring Method / 分析化学

13 C isotopic abundance of intracellular free amino acid with a characteristic of fast- turnover can quickly reflect changes in intracellular metabolic state. But the concentration of intracellular free amino acid is low, the existed 13 C isotope detection method based on GC-MS can not satisfy the requirement with full scan mode. In this study, the selected ion monitoring method was used to detect accuracy higher likelihood of analysis of 13 C isotopic abundance of free intracellular amino acid. First, in the full scan mode we analyzed of the fracture law of different amino acids, found the feature corresponding to each amino acid fragments, and established 16 kinds of free intracellular amino acids characteristic fragment library. Then using this characteristic fragment library, only specific m/z signal was detected in sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards showed that the signal-to-noise ratio, measurement precision and accuracy were improved by 17, 2. 0 and 3. 8 times compared with the full scan mode. In the analysis of coenzyme Q10 producing strains of samples, this method was successfully used to detect isotopic abundance of 8 kinds of free intracellular amino acids. This method plays an important role in the detection of 13 C isotopic abundance of the intracellular free amino acid in cell metabolism research.

Role of phosphorylation of MARCKS-PSD in the secretion of MUC5AC induced by cold temperatures in human airway epithelial cells / 中南大学学报(医学版)

OBJECTIVE@#To construct phosphorylation sites domain (PSD) mutant of myristoylated alaninerich C kinase substrate (MARCKS) and explore the role of transient receptor potential melastatin 8 cation channels (TRPM8) and MARCKS in cold-induced synthesis and exocytosis of mucin (MUC) 5AC.@*METHODS@#Human placental cDNA was used as a template to amplify the full coding region of MARCKS cDNA by PCR. Ser159, Ser 163, Ser 167, Ser 170 in the PSD were mutated to aspartic acids by an overlap PCR method. The resultant PSD mutant cDNA and the wild-type MARCKS cDNA were each subcloned into a mammalian expression vector pcDNA3.0. Recombinant constructs were confirmed by restriction enzyme digestion analysis and DNA sequencing. In intervention experiments, cells were pretreated with the TRPM8 channel antagonist BCTC and transfected with MARCKS-PSD mutant cDNA, and thereafter cold stimulation was applied. The levels of MUC5AC were measured by immunofluorescence and ELISA to clarify the roles of TRPM8 and PSD mutant on the synthesis and secretion of MUC5AC induced by cold, respectively.@*RESULTS@#Restriction enzyme digestion analysis and DNA sequencing revealed that the pcDNA3.0- MARCKS and pcDNA3.0-MARCKS-PSD mutants were successfully constructed. The levels of intracellular and secreted MUC5AC of cold treated group were significantly higher than those of control group (P0.05).@*CONCLUSION@#TRPM8 and phosphorylation of MARCKS-PSD mediates the cold-induced exocytosis of MUC5AC by airway epithelial cells.

Discussion on the English teaching of diagnostics to international students / 中华医学教育探索杂志

With the development of internationalization of medical education,teaching specialty in English has been a new task for many general medical colleges and universities.Many problems need to be analyzed and solved.In the process of teaching international students diagnostics,teaching mode has been actively explored.The management of teaching,the foundation of teaching team,the selection of teaching materials and reformation of teaching mode are the key points that affect the teaching quality directly.

Detection of interleukin-10 in alveolar macrophage and peripheral blood monocyte from lung cancer patients and its clinical significance / 中国肺癌杂志

<p><b>BACKGROUND</b>To investigate the ability of alveolar macrophage (AM) and peripheral blood monocyte (PBMC) from lung cancer patients to produce Interleukin-10 (IL-10) and to evaluate the local and general cellular immune state of lung cancer patients.</p><p><b>METHODS</b>AM and PBMC were obtained from 57 patients with lung cancer, 33 patients with benign pulmonary diseases and 12 healthy volunteers. IL-10 in the culture supernatants was measured quantitatively by ELISA.</p><p><b>RESULTS</b>(1) Elevated levels of IL-10 produced by PBMC were found in lung cancer group with respect to healthy volunteers and patients with benign pulmonary diseases (148.60±35.56 vs 93.83±9.22 and 108.91±15.95 ng×L⁻¹ respectively) (P < 0.001). The level of IL-10 by AM from lung cancer patients was 132.06±30.42 ng×L⁻¹, which was remarkably higher than that of healthy volunteers (92.67±11.22 ng×L⁻¹) and benign pulmonary diseases (94.39±10.04 ng×L⁻¹) (P < 0.001). (2) The level of IL-10 produced by PBMC from lung cancer group of stage IV was significantly higher than that of stage I+II (178.33±13.52 vs 131.57±25.35 ng×L⁻¹) (P < 0.001). AM from lung cancer patients of stage III and IV produced more IL-10 than that of stage I+II did (150.13±15.57 and 160.50± 18.75 vs 117.05±28.12 ng×L⁻¹ respectively) (P < 0.001); PBMC from small cell lung cancer patients produced higher level of IL-10 than squamous cell carcinoma and adenocarcinoma did (194.83±23.88 vs 140.37± 27.00 ng×L⁻¹ and 136.50±27.39 ng×L⁻¹ respectively) (P < 0.01). AM from small cell lung cancer patients produced higher levels of IL-10 than squamous cell carcinoma and adenocarcinoma did (165.33±23.78 vs 127.74±26.19 ng×L⁻¹ and 120.30±29.66 ng×L⁻¹ respectively) (P < 0.01); Size of mass and performance status greatly affected the level of IL-10; IL-10 level of lung cancer patients with survival time more than 2 years was remarkably lower than that of patients with survival time less than 2 years (P < 0.01).</p><p><b>CONCLUSIONS</b>Higher level of IL-10 presents both in local and general body of lung cancer patients. IL-10 may play an important role in deterioration of lung cancer. Detection of IL-10 level may be helpful to evaluate cellular immunity and predict prognosis of lung cancer patients.</p>

Production of IL-10 and IL-12 by AM and PBMC from lung cancer patients / 重庆医科大学学报

Objective:To investigate the production of IL-10 and IL-12 by alveolar marcrophage and peripheral blood mononuclear cell from lung cancer patients,to evaluate the cellular immune state,and to provide justification for IL-12 therapy in the fields of Lung cancer.Methods:AM and PBMC were obtained from 57 patients with Lung cancer,33 patients with benign pulmonary disease,and 12 normal control.IL-10 and IL-12 in the culture supernatants were measured quantitatively by ELISA.Results:(1) Higher levels of IL-10 were found in Lung cancer group than that in NC and BPD group.IL-12 showed significantly lower levels than that in NC and BPD group.(2) The levels of IL-10 had negative correlation with that of IL-12 in Lung cancer group.(3) The levels of IL-10 in Lung cancer group of stage Ⅲ,Ⅳ were significantly higher than that of stage Ⅰ～Ⅱ.The levels of IL-12 in Lung cancer group of stage Ⅳ were significantly lower than that of stage Ⅰ～Ⅱ.(4) Higher levels of IL-10 were found in SCLC group than that in NSCLC group.The levels of IL-12 from PBMC and AM in SCLC group were lower than that in adenocarcinoma,adenocarcinoma and squamous carcinoma groups separately.Conclusion:(1) results suggest the defect of cell-mediated immunity function presents both in local and general immune response in Lung cancer patients.(2)The basal levels of IL-10 and IL-12 are related to the stage and extent of malignancy.

Exploration of clinical diagnostic value of measuring sulfate glycosaminoglycan segment and antibody to tuberculosis in patients with hydrothorax / 重庆医科大学学报

Objective:To explore the clinical value of detection of sulfated glycosaminoglycan(GAG) segment and IgG antibody to tuberculosis(TB-IgG) to diagnose tuberculous pleuritis and malignant pleural effusion.Methods:Using the lung tumor antigen(LTA) diagnostic kit and tuberculosis antibody colloidal gold diagnostic kit to detecte the GAG and TB-IgG of serum and pleural fluid separately of 357 patients suffering from hydrothorax.We proceeded a retrospective study and then had statistical evidence.Results:The positive rate and specificity of GAG of pleural effusion of lung cancer patients were 80.3%(106/132),88.4%,and that of serum were 54.5%(72/132),90.2%.The positive rate and specificity of TB-IgG of pleural effusion of tuberculous pleuritis were 66.2%(102/154),86.2%,and that of serum were 33.8%(52/154),83.7%.The specificity of GAG,combined with TB-IgG by measuring the pleural fluid of tuberculous pleuritis and malignant pleural effusion would increase to 97.0% and 98.2% separately.Conclusion:Detection of GAG and TB-IgG of pleural fluid is helpul for diagnosis of tuberculous pleuritis and malignant pleural effusion.GAG combined with TB-IgG can act as differential diagnosis marker of tuberculous pleuritis and malignant pleural effusion for its higher specificity.

The alert role of Clara cell secretory protein in serum from patients in the developing course of acute respiratory distress syndrome / 重庆医科大学学报

Objective:To search for a prophetic marker reflecting the progress of acute respiratory distress syndrome (ARDS).Methods:The serum was obtained from normal controls and patients at difierent stages of ARDS:systemic inflammatory response syndrome (SIRS),acute lung injury(ALI),ARDS and just before dying.Clara cell secretory protein (CC16) in serum was detected using enzyme-linked immunoadsordent assay (ELISA).Results:In the early stage of inflammatory reaction,that is,stage of SIRS,the CC16 levels in serum were higher than normal controls ( P 0.01).Conclusion:The determination of CC16 in serum is a sensitive marker reflecting the integrity of alveolar epithelium and vascular endothelial cell.The alteration of CC16 is presented in the early period of acute inflammatory reaction.The alteration of CC16 is earlier than that of blood gas analysis.CC16 may serve as a helpful marker to foresee and evaluate prognosis of ARDS in early stage.