Burkholderia pseudomallei infection manifests as mediastinal/hilar lymphadenopathy: A case report

Rationale: This case report presents the diagnosis and etiology of hilar/mediastinal lymphadenopathy in a male patient. Patient concerns: A 49-year-old man presented with fever and dyspnea after physical exertion. Diagnosis: The patient was diagnosed with melioidosis by cultivation of lymph node aspirate on blood agar using the VITEK 2 compact system. Interventions: The patient was treated with ceftazidime intravenously, combined with trimethoprim/sulfamethoxazole orally for 1 week. Once the patient was discharged, he began a 12-week course of trimethoprim/sulfamethoxazole. Outcomes: The patient recovered after treatment with ceftazidime and trimethoprim/sulfamethoxazole. Conclusions: Melioidosis is an infectious disease that mainly occurs in tropical regions. It can cause severe sepsis and pneumonia, and the infection in some patients may become chronic. Endobronchial ultrasound-transbronchial needle aspiration is a useful technique in the diagnosis of patients with hilar/mediastinal lymphadenopathy.

TRPM8 is overexpressed in the respiratory tract of steroid-naive asthma patients

Background:TRPM8 is a member of TRP channels family known for its sensitivity to cool temperature, increased osmotic pressure, particulate matter, cigarette smoke and products of oxidative stress. This cation channel is widely expressed in the respiratory tract including airway epithelium and vagal nerve endings where it can mediate inflammatory and secretory responses. Previous studies have reported the increased expression of TRPM8 in the respiratory tract of COPD patients.Objective:To investigate TRPM8 expression in nasal epithelium and induced sputum of asthma patients and to estimate its relation to airway hyperresponsiveness induced by cold air.Methods:The study enrolled 43 subjects with mean age of (39.8±1.76) years including 35 patients with mild-to-moderate asthma and 8 patients with chronic bronchitis. Among the asthma patients 43% were steroid-naïve. Lung function was measured by standard spirometry before and after bronchoprovocation challenge with 3 min isocapnic cold air (-20 °C) hyperventilation. Cold air hyperresponsiveness was diagnosed in case of FEV1 falling by 10% from baseline. TRPM8 expression was measured by indirect flow cytometry in nasal brushings and induced sputum. Nasal epithelium cells were gated after exclusion of dead cells and staining with anti- cytokeratin 19 antibodies. Induced sputum was processed with dithiothreitol, filtered and stained for CD45 and cell viability. Extracellular expression of TRPM8 was detected by staining with primary unconjugated anti-TRPM8 antibodies (Alomone Labs) and secondary antibodies labeled with Alexa Fluor 488 (Abcam). Expression level was calculated as normalized median fluorescence intensity (nMFI) and percent of positively stained cells (%Pos). The data are presented as median, lower and upper quartiles (Me (Q1; Q3)).Results:TRPM8 protein was detected on the epithelial cells and sputum macrophages. The expression levels of TRPM8 were significantly correlated in these cells (nMFI R=0.41, P=0.04; %Pos R=0.53, P=0.008). Expression of TRPM8 was increased both in nasal epithelium and macrophages of steroid-naïve asthma patients as compared to the patients receiving maintenance therapy or patients with chronic bronchitis. nMFI values for macrophages were 2.1 (1.95; 2.79), 1.3 (1.10; 1.91) and 1.2 (1.06; 1.35), respectively (P=0.001). %Pos values for macrophages were 38.5 (13.1; 51.1)%, 11.6 (2.55; 20.1)% and 8.0 (3.20; 9.20)%, respectively (P=0.01). nMFI and %Pos values for the epithelium kept the same trend but the differences were mostly insignificant. In addition, we found a correlation between FEV1 in response to cold air hyperventilation challenge and TRPM8 expression (macrophages %Pos R=-0.43, P=0.02; epithelium %Pos R=- 0.43, P=0.08). Cold airway hyperresponsiveness was accompanied by higher TRPM8 expression on macrophages (21.6 (17.1; 40.7)%vs.11.6 (7.45; 16.55)%, P=0.03) and nasal epithelium (4.2 (3.30; 5.40)%vs.2.8 (1.10; 4.30)%, P=0.2). Cold airway hyperresponsiveness was not affected by maintenance therapy in our patients.Conclusions:TRPM8 is overexpressed in steroid-naïve asthma patients as well as in asthma patients with cold airway hyperresponsiveness and may be implicated in asthma pathogenesis. The utility of TRPM8 as a diagnostic biomarker or therapeutic target in chronic obstructive airway diseases is of great interest.

Limonin inhibits the PM2.5 inhalation-induced airway inflammation and mucus hypersecretion in rats / 基础医学与临床

Objective To investigate the inhibitory effect of limonin on airway inflammation and mucus hypersecre-tion. Methods The experiment was divided into three groups(n=10),namely blank control group,PM2.5 group (the rat models of chronic airway inflammation were established by aerosolized PM2.5 suspension) and PM2.5+limonin group(intervening with the extract from tangerine peel). mRNA,protein of inflammatory cytokines,mucin (MUC) and TAS2Rs were measured by ELISA,RT-PCR and Western blot respectively. Results The mRNA and protein expression of IL-1β, CINC-1 and MUC5AC and MUC5B in PM2.5 group were significantly higher than those in control group(P<0.05),and the expression of MUC5AC protein in broncho alveolar lavage fluid(BALF) was increased. Compared with PM2.5 stimulated group,mRNA and protein of TAS2R14 in PM2.5+limonin inter-vented group were significantly higher (P<0.05). Moreover, the expression of IL-1β, CINC-1 and MUC5AC, MUC5B was lower than PM2.5 group (P<0.05).While the expression of MUC5B was mainly increased in BALF.Conclusions The production of mucin can be inhibited by aerosolized limonin, meanwhile the secretion of mucin also can be promoted.This effect is achieved by activating the expression of TAS2Rs in the lungs, which enhances the anti-inflammatory effect of airway inflammation.

Role of Wnt Inhibitory Factor-1 in Inhibition of Bisdemethoxycurcumin Mediated Epithelial-to-Mesenchymal Transition in Highly Metastatic Lung Cancer 95D Cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Bisdemethoxycurcumin (BDMC) is an active component of curcumin and a chemotherapeutic agent, which has been suggested to inhibit tumor growth, invasion and metastasis in multiple cancers. But its contribution and mechanism of action in invasion and metastasis of non-small cell lung cancer (NSCLC) are not very clear. Therefore, we tried to study the effects of BDMC on regulation of epithelial-to-mesenchymal transition (EMT), which is closely linked to tumor cell invasion and metastasis.</p><p><b>METHODS</b>In this study, we first induced transforming growth factor-β1 (TGF-β1) mediated EMT in highly metastatic lung cancer 95D cells. Thereafter, we studied the effects of BDMC on invasion and migration of 95D cells. In addition, EMT markers expressions were also analyzed by western blot and immunofluorescence assays. The contribution of Wnt inhibitory factor-1 (WIF-1) in regulating BDMC effects on TGF-β1 induced EMT were further analyzed by its overexpression and small interfering RNA knockdown studies.</p><p><b>RESULTS</b>It was observed that BDMC inhibited the TGF-β1 induced EMT in 95D cells. Furthermore, it also inhibited the Wnt signaling pathway by upregulating WIF-1 protein expression. In addition, WIF-1 manipulation studies further revealed that WIF-1 is a central molecule mediating BDMC response towards TGF-β1 induced EMT by regulating cell invasion and migration.</p><p><b>CONCLUSIONS</b>Our study concluded that BDMC effects on TGF-β1 induced EMT in NSCLC are mediated through WIF-1 and elucidated a novel mechanism of EMT regulation by BDMC.</p>

Autophagy Accompanied with Bisdemethoxycurcumin-induced Apoptosis in Non-small Cell Lung Cancer Cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the effects of bisdemethoxycurcumin (BDMC) on non-small cell lung cancer (NSCLC) cell line, A549, and the highly metastatic lung cancer 95D cells.</p><p><b>METHODS</b>CCK-8 assay was used to assess the effect of BDMC on cytotoxicity. Flow cytometry was used to evaluate apoptosis. Western blot analysis, electron microscopy, and quantification of GFP-LC3 punctuates were used to test the effect of BDMC on autophagy and apoptosis of lung cancer cells.</p><p><b>RESULTS</b>BDMC inhibited the viability of NSCLC cells, but had no cytotoxic effects on lung small airway epithelial cells (SAECs). The apoptotic cell death induced by BDMC was accompanied with the induction of autophagy in NSCLC cells. Blockage of autophagy by the autophagy inhibitor 3-methyladenine (3-MA) repressed the growth inhibitory effects and induction of apoptosis by BDMC. In addition, BDMC treatment significantly decreased smoothened (SMO) and the transcription factor glioma-associated oncogene 1 (Gli1) expression. Furthermore, depletion of Gli1 by siRNA and cyclopamine (a specific SMO inhibitor) induced autophagy.</p><p><b>CONCLUSION</b>Aberrant activation of Hedgehog (Hh) signaling has been implicated in several human cancers, including lung cancers. The present findings provide direct evidence that BDMC-induced autophagy plays a pro-death role in NSCLC, in part, by inhibiting Hedgehog signaling.</p>

Study on three different species tibetan medicine sea buckthorn by 1H-NMR-based metabonomics / 中国中药杂志

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.

Synthesis and antimicrobial evaluation of coumarin-based benzotriazoles and their synergistic effects with chloromycin and fluconazole / 药学学报

A series of new coumarin-based benzotriazole derivatives were successfully synthesized via a multi-step sequence of cyclization, etherification and N-alkylation, and were confirmed by 1H NMR, IR, MS spectra as well as elemental analyses. All these synthesized coumarin compounds were evaluated for in vitro antimicrobial activities against four Gram-positive bacteria, four Gram-negative bacteria and three fungi by two fold serial dilution technique. The bioactive assay showed that all these prepared coumarin benzotriazoles could inhibit the growth of the tested bacterial and fungal strains. Title compounds 11a-11e and 13a-13c were more active than chloromycin on Proteus vulgaris ATCC 6896. Coumarin benzotriazoles 11a and 11b displayed comparable antibacterial efficacy against Staphylococcus aureus ATCC 25923 and Micrococcus luteus ATCC 4698 in comparison with reference drug chloromycin. Compared to fluconazole, compounds 11a-11d displayed stronger inhibition on Aspergillus fumigatus ATCC 96918. Moreover, coumarin-based benzotriazoles in combination with antibacterial chloromycin or antifungal fluconazole, showed notable antimicrobial efficacy with less dosage and broader antimicrobial spectrum. More importantly, fluconazole-insensitive A. fumigatus and methicillin-resistant Staphylococcus aureus N 315 (MRSA) were sensitive to these combined drugs.

Molecular mechanism of interleukin-13-induced mucus hypersecretion in rat airway / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of interleukin-13 (IL-13) on mucus secretion in vivo and the possible mechanism.</p><p><b>METHODS</b>The SD rats were randomly divided into control group, IL-13 group and IL-13 plus SP600125 group. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2) and the level of MUC5AC in the lung tissues were examined using Western blotting. RT-PCR was performed to examine the mRNA level of STAT4 and STAT6, and electrophoretic mobility shift assays (EMSA) was used to detect the DNA-binding activities of Forkhead box a2 (FOXA2) and activator protein-1 (AP-1).</p><p><b>RESULTS</b>IL-13 caused a significant increase in MUC5AC and p-JNK1/2 expression, but did not affect the phosphorylation of ERK1/2. The expression of MUC5AC was attenuated after treatment with SP600125. A significant increase in STAT6 was observed in IL-13 group compared with that in the control group, whereas the expression of STAT4 mRNA was not significantly affected. The DNA-binding activity of FOXA2 was down-regulated after IL-13 exposure, which did not affect the DNA-binding activity of AP-1.</p><p><b>CONCLUSION</b>IL-13 down-regulates mucus secretion via STAT6-FOXA2 pathway in vitro.</p>

Effects of Scutellarin on MUC5AC Mucin Production Induced by Human Neutrophil Elastase or Interleukin 13 on Airway Epithelial Cells

Scutellarin is a flavonoid extracted from a traditional Chinese herb, Erigeron breviscapus. The present study investigated the effect of scutellarin on MUC5AC mucin production and the possible mechanism. Human bronchial epithelial 16 (HBE16) cells were pretreated with scutellarin for 60 min, and then exposed to human neutrophil elastase (HNE) or interleukin (IL)-13 for 12 hr. RT-PCR and ELISA were performed to measure the amount of MUC5AC mucin production. The results showed that scutellarin inhibited MUC5AC expression both in mRNA and protein level induced by HNE in a concentration-dependent manner. However, scutellarin failed to inhibit MUC5AC mucin production induced by IL-13. To investigate the intracellular mechanisms associated with the effect of scutellarin on MUC5AC mucin production, western blotting was carried out to examine the phosphorylation of protein kinase C (PKC), signal transducer and activator of transcription 6 (STAT6) and extracellular signal-regulated kinase 1/2 (ERK1/2). The phosphorylation of PKC and ERK1/2 was attenuated after treatment with scutellarin, whereas STAT6 was not significantly affected. Therefore, it is suggested that scutellarin down-regulates MUC5AC mucin production on HBE16 cells via ERK-dependent and PKC-dependent pathways.

Effect of nordy on FPR function of malignant human glioma cell line U87 / 药学学报

Nordy is a synthesized chrial compound. To investigate the effects of nordy (25 - 100 micromol x L(-1)) on the function of formylpeptide receptor (FPR) of malignant human glioma cells, human glioblastoma cell line U87 was used to detect its proliferation, migration, calcium mobilization, vascular endothelial growth factor (VEGF) mRNA and protein levels after activation of FPR by its agonist N-formyl-methionyl-leucyl-phenylalanine (fMLF). Cell proliferation, migration ability, VEGF mRNA, VEGF protein and calcium mobilization were evaluated by cell counting, chemotaxis assay, RT-PCR, ELISA and spectrometry. Nordy (50 - 100 micromol x L(-1)) potently inhibited the proliferation, migration and calcium mobilization of U87 cells induced by fMLF (P < 0.05). Moreover, 100 micromol x L(-1) nordy showed a significantly impaired VEGF mRNA expression and protein secretion induced by fMLF (P < 0.05). Nordy could inhibit FPR functioning in glioma cell proliferation, migration and angiogenesis, which might be a possible mechanism of its anti-cancer effects.

Effect of nordy on biological behaviors of malignant glioma cell line U87MG and the analysis of differential expression proteome / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore effects of nordy on biological behaviors of human malignant glioblastoma cell line U87MG in vitro and transplanted tumor in vivo, and to identify the differential proteome upon Nordy induced differentiation.</p><p><b>METHODS</b>Glioblastoma U87MG cells were induced to differentiate by synthetic lipoxygenase inhibitor, Nordy. The drug was also given via peritoneal injection to nude mice (27 mg/kg body weight) bearing orthotopic transplanted tumors of U87MG cells in the brain. The tumor volumes and GFAP expression were measured. Total proteins of U87MG cells after Nordy treatment were analysed by two-dimensional gel electrophoresis. PDQuest 7.1 computer software was used to compare protein profiles of the treated cells with that of untreated control. Differentially expressed proteins were then selected and characterized by matrix assisted laser desorption ionization-time of flight-mass spectrometry. The functional aspects of these proteins were analyzed by bioinformatics.</p><p><b>RESULTS</b>Nordy suppressed both the proliferation of U87MG cells in vitro and the tumor growth of orthotopic transplanted tumors in vivo (P < 0.01). The differentially expressed proteins induced by Nordy included proliferation-associated gene A, alternative splicing factor ASF-3, eukaryotic translation initiation factor 5A, coffilin 1 (non-muscle), beta galactoside binding lectin, glyceraldehyde-3-phosphate dehydrogenase, enolase 1 and an unknown protein.</p><p><b>CONCLUSIONS</b>Nordy promotes the differentiation of glioblastoma cells, by which it may serve as a therapeutic agent. Various proteins identified during Nordy-induced differentiation are involved in the cell proliferation, metabolism, differentiation, apoptosis and gene transcription.</p>

Roles of theaflavins in inflammatory airway mucus hypersecretion / 上海第二医科大学学报

Objective To investigate the roles of theaflavins(TFs) in airway mucus hypersecretion induced by human neutrophil elastase(HNE). Methods Human lung adenocarcinoma cell A549 was stimulated by HNE for mucus hypersecretion,and TF monomers(TF1,TF2 and TF3) were used for intervention.The effects of TF monomers on viability of A549 cells were examined by MTT method.After the effective doses of TFs were determined,A549 cells were divided into 4 groups for experiment.In control group,A549 cells were cultured with serum-free medium.In HNE treatment group,A549 cells were treated with HNE(50 nmol/L) for 24 h.In TF monomer intervention groups,A549 cells were pre-treated with TF1,TF2 or TF3(50,100 or 200 ?g/mL) for 24 h,and were then treated with HNE for another 24 h.In AG1478 intervention group,A549 cells were pre-treated with AG1478(5 ?mol/L),an epidermal growth factor receptor blocker for 30 min,and were then treated with HNE for another 24 h.The changes in mucin(MUC) after treatment by different doses of TF1,TF2 and TF3,and by TF3(200 ?g/mL) for different time(12 h,24 h and 36 h) were detected.The changes in MUC5AC mRNA expression and MUC5AC protein expression were detected by RT-PCR and ELISA,respectively. Results The MUC5AC mRNA expression and protein expression in HNE treatment group were significantly higher than those in control group(P

Analysis of Cis-acting Elements on Respiratory Mucin MUC5AC Expression and Regulation in Pulmonary A549 Cell Line / 中国生物工程杂志

Objective:To study the mechanisms of neutrophil elastase (NE) induced expression of respiratory mucin MUC5AC. Methods:Using gene recombination techniques,four luciferase reporter gene plasmids containing different length of human MUC5AC gene promoter were constructed.Site-directed mutagenesis technique was used to establish mutants of Sp-l and NF-?B site in MUC5AC gene promoter; the relative luciferase activities were detected in the transfected human pulmonary A549 cells. Results:Series of luciferase reporter gene containing different sequences of human MUC5AC promotor were constructed successfully.NE could increase the expression of luciferase reporter gene plasmid containing -1300bp,-689bp and-324bp version of MUC5AC promoter in the transfected A549 cells (P