Publications by authors named "Aihua Bao"

20 Publications

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Simultaneous and rapid determination of 12 tyrosine kinase inhibitors by LC-MS/MS in human plasma: Application to therapeutic drug monitoring in patients with non-small cell lung cancer.

J Chromatogr B Analyt Technol Biomed Life Sci 2021 Jun 1;1175:122752. Epub 2021 May 1.

Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, PR China. Electronic address:

In recent years, more than 50 tyrosine kinase inhibitors (TKIs) was indicated against numerous cancers, especially outstanding advantages in the treatment of non-small cell lung cancer (NSCLC), and several studies have shown that therapeutic drug monitoring (TDM) of TKIs can improve treatment efficacy and safety. The present study aimed to develop and validate a LC-MS/MS method for the TDM of 12 TKIs (gefitinib, erlotinib, afatinib, dacomitinib, icotinib, osimertinib, crizotinib, ceritinib, alectinib, dabrafenib, trametinib, anlotinib) in patients with NSCLC. The analytes of interest and internal standard were extracted from human plasma. Salting-out assisted liquid-liquid extraction (SALLE) with 5 M ammonium acetate solution was optimized for method validation and compared to simple protein precipitation (PPT). Chromatographic separation was conducted on Waters X bridge C18 column (100 × 4.6 mm, 3.5 μm) using a gradient elution of acetonitrile/5mM ammonium acetate in pure water with 0.1% (v/v) formic acid at 40 °C within 6 min. The total flow was maintained at 1 mL/min, 30% of the post column flow was split into the mass spectrometer and the rest to waste via a 3-way tee. The mass analysis was performed by positive ion electrospray ionization (ESI) in multiple-reaction monitoring (MRM) mode. The assay was validated based on the guidelines on bioanalytical methods by FDA. This quantification method was proved to be satisfactory in selectivity, accuracy, precision, linearity (r > 0.995), recovery, matrix effect and stability and the accuracy was further assessed in plasma with a degree of hemolysis of 4%. The described method to simultaneously quantify the 12 selected anticancer drugs in human plasma was successfully validated and applied to routine TDM of gefitinib, erlotinib, icotinib, osimertinib, crizotinib and anlotinib in cancer patients. TKIs plasma monitoring helps to individualize dose adjustment and manage adverse effects in NSCLC patients.
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http://dx.doi.org/10.1016/j.jchromb.2021.122752DOI Listing
June 2021

Clinical features in coronavirus disease 2019 (COVID-19) patients with early clearance and prolonged shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA.

Ann Transl Med 2021 Apr;9(8):665

Department of Respiratory and Critical Care Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Background: Since the outbreak of coronavirus disease 2019 (COVID-19), the pattern of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA shedding has not been well characterized.

Methods: In our study, 652 patients in Wuhan Designated Hospital were recruited, and their clinical and laboratory findings were extracted and analyzed.

Results: The median duration of SARS-CoV-2 RNA detection was 23 days [interquartile range (IQR), 18 days] from symptom onset. Compared to patients with early viral RNA clearance (<23 days after illness onset), we found that patients with late viral RNA clearance (≥23 days) had a higher proportion of clinical features, as follows: symptoms, including fever, dry cough, and sputum production; comorbidities, including hypertension, chronic kidney disease, uremia, chronic liver disease, anemia, hyperlipidemia, and bilateral lung involvement; complications, such as liver injury; delayed admission to hospital; laboratory parameters at baseline, including higher eosinophils, uric acid, cholesterol, triglycerides, and lower hemoglobin; and less treatment with arbidol, chloroquine, or any antivirals. After generalized linear regression, prolonged SARS-CoV-2 RNA shedding was independently associated with younger age; delayed admission to hospital; symptoms including fever, shivering, and sputum production; comorbidities including hypertension, diabetes, cardiovascular disease, anemia, hyperlipidemia, uremia, and lung involvement; and higher alanine aminotransferase (ALT), uric acid, and cholesterol levels at baseline.

Conclusions: In conclusion, the factors mentioned above are associated with the negative conversion of SARS-CoV-2 RNA. A deeper insight into virological dynamics will be helpful for establishing patient discharge and quarantine release criteria.
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http://dx.doi.org/10.21037/atm-21-445DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106067PMC
April 2021

Administration route governs the therapeutic efficacy, biodistribution and macrophage targeting of anti-inflammatory nanoparticles in the lung.

J Nanobiotechnology 2021 Feb 25;19(1):56. Epub 2021 Feb 25.

School of Biomedical Engineering, Tianjin Medical University, Tianjin, 300070, China.

Background: Uncontrolled inflammation is a central problem for many respiratory diseases. The development of potent, targeted anti-inflammatory therapies to reduce lung inflammation and re-establish the homeostasis in the respiratory tract is still a challenge. Previously, we developed a unique anti-inflammatory nanodrug, P12 (made of hexapeptides and gold nanoparticles), which can attenuate Toll-like receptor-mediated inflammatory responses in macrophages. However, the effect of the administration route on its therapeutic efficacy and tissue distribution remained to be defined.

Results: In this study, we systematically compared the effects of three different administration routes [the intratracheal (i.t.), intravenous (i.v.) and intraperitoneal (i.p.)] on the therapeutic activity, biodistribution and pulmonary cell targeting features of P12. Using the LPS-induced ALI mouse model, we found that the local administration route via i.t. instillation was superior in reducing lung inflammation than the other two routes even treated with a lower concentration of P12. Further studies on nanoparticle biodistribution showed that the i.t. administration led to more accumulation of P12 in the lungs but less in the liver and other organs; however, the i.v. and i.p. administration resulted in more nanoparticle accumulation in the liver and lymph nodes, respectively, but less in the lungs. Such a lung favorable distribution was also determined by the unique surface chemistry of P12. Furthermore, the inflammatory condition in the lung could decrease the accumulation of nanoparticles in the lung and liver, while increasing their distribution in the spleen and heart. Interestingly, the i.t. administration route helped the nanoparticles specifically target the lung macrophages, whereas the other two administration routes did not.

Conclusion: The i.t. administration is better for treating ALI using nanodevices as it enhances the bioavailability and efficacy of the nanodrugs in the target cells of the lung and reduces the potential systematic side effects.
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http://dx.doi.org/10.1186/s12951-021-00803-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7905431PMC
February 2021

Correction to: Inducible expression of heat shock protein 20 protects airway epithelial cells against oxidative injury involving the Nrf2-NQO-1 pathway.

Cell Biosci 2020 Dec 19;10(1):145. Epub 2020 Dec 19.

Department of Respiratory and Critical Care Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai, 200080, China.

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http://dx.doi.org/10.1186/s13578-020-00510-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7749981PMC
December 2020

Inducible expression of heat shock protein 20 protects airway epithelial cells against oxidative injury involving the Nrf2-NQO-1 pathway.

Cell Biosci 2020 19;10:120. Epub 2020 Oct 19.

Department of Respiratory and Critical Care Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai, 200080 China.

Background: Heat shock protein (HSP) 20 is a molecular chaperone that exerts multiple protective functions in various kinds of tissues. However, the expression of HSP20 and its specific functions in airway epithelial cells (AECs) remain elusive.

Results: In current study, we first confirmed the inducible expression of HSP20 in mouse AECs and in a human bronchial epithelial cell line BEAS-2B cells, under different oxidant stressors. Then by establishing a HSP20-abundant mouse model with repeated low-level-ozone exposures and stimulating this model with a single high-level ozone exposure, we found that the HSP20 abundance along with its enhanced phosphorylation potentially contributed to the alleviation of oxidative injuries, evidenced by the decreases in the bodyweight reduction, the BAL neutrophil accumulation, the AECs shedding, and the BAL concentrations of albumin and E-cadherin. The biological function of HSP20 and its molecular mechanisms were further investigated in BEAS-2B cells that were transfected with -, unphosphorylatable or empty vector plasmids prior to the stimulation of HO, of which its oxidant capacity has been proved to be similar with those of ozone in an air-liquid culture system. We found that the HO-induced intracellular ROS level and the early cell apoptosis were attenuated in the but not transfected cells. The intracellular expression of NQO-1 (mRNA and protein) and the intranuclear content of Nrf2 were significantly increased in the transfected cells but not in the - and empty vector-transfected cells after the stimulation of HO.

Conclusions: The inducible expression of HSP20 in AECs by oxidative stress exerts protective roles against oxidative damages, which may involve the activation of the Nrf2-NQO-1 pathway.
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http://dx.doi.org/10.1186/s13578-020-00483-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7574176PMC
October 2020

Adenovirus vector-mediated YKL-40 shRNA attenuates eosinophil airway inflammation in a murine asthmatic model.

Gene Ther 2021 Apr 12;28(3-4):177-185. Epub 2020 Oct 12.

Department of Respiratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Recent studies have revealed that YKL-40 is involved in the pathogenesis of asthma. However, its specific mechanism remains unclear. The present study aims to investigate the effect of adenovirus vector-mediated YKL-40 short hairpin RNA (shRNA) on regulation of airway inflammation in a murine asthmatic model. Mice were assessed for airway hyperresponsiveness (AHR), total leukocytes and the percentage of eosinophil cells in bronchoalveolar lavage fluid (BALF). YKL-40 mRNA and protein expression levels were detected using quantitative real-time PCR and western blot assays. Enzyme-linked immunosorbent assay (ELISA) was used to detect YKL-40 and eosinophil-related chemokine expression levels in BALF and serum. Lung histology analyses were performed to evaluate the degree of inflammatory cell infiltration around the airway and airway mucus secretion.YKL-40 shRNA significantly inhibited the YKL-40 gene expression in asthmatic mice. In addition, YKL-40 shRNA alleviated eosinophilic airway inflammation, AHR, airway mucus secretion and decreased the levels of YKL-40 in BALF and serum in a murine asthmatic model. The levels and mRNA expression of IL-5, IL-13 in asthmatic mice lung tissues, eotaxin, and GM-CSF in BALF and serum significantly decreased. Bone marrow signaling molecules including IL-5, eotaxin, and GM-CSF were correlated with decreased levels of YKL-40. The study reveals that YKL-40 could be involved in asthma inflammation by altering bone marrow signaling molecules. YKL-40 gene RNA interference could provide new therapeutic strategies for asthma.
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http://dx.doi.org/10.1038/s41434-020-00202-0DOI Listing
April 2021

Lung Ultrasound Score in Evaluating the Severity of Coronavirus Disease 2019 (COVID-19) Pneumonia.

Ultrasound Med Biol 2020 11 24;46(11):2938-2944. Epub 2020 Jul 24.

Department of Emergency and Critical Care, Shanghai General Hospital of Shanghai Jiaotong University, Shanghai, China. Electronic address:

The purpose of this study is to observe the potential of lung ultrasound in evaluating the severity of coronavirus disease 2019 (COVID-19) pneumonia. Lung ultrasound was performed in ten zones of the patients' chest walls. The features of the ultrasound images were observed, and a lung ultrasound score (LUS) was recorded. The ultrasound features and scores were compared between the refractory group (PaO/FiO ≤ 100 mm Hg or on extracorporeal membrane oxygenation) and the non-refractory group. The prediction value of the LUS was studied by receiver operating characteristic (ROC) curve analysis. In total, 7 patients were enrolled in the refractory group and 28 in the non-refractory group. B-line patterns and shred signs were the most common signs in all patients. Patients in the refractory group had significantly more ground-glass signs (median 6 [interquartile range {IQR}, 2.5-6.5] vs. median 0 [IQR, 0-3]), consolidation signs (median 1 [IQR, 1-1.5] vs. median 0 [IQR, 0-3]) and pleural effusions (median 5 [IQR, 1.5-6] vs. median 0 [IQR, 0-0.25]). The LUS was significantly higher in the refractory group (33.00 [IQR 27.50-34.00] vs. 25.50 [IQR 22.75-30.00]). The ROC of the LUS showed a cutoff score of 32 with a specificity of 0.893 and a sensitivity of 0.571 in diagnosing refractory respiratory failure among patients. In COVID-19 patients, lung ultrasound is a promising diagnostic tool in diagnosing patients with refractory pneumonia.
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http://dx.doi.org/10.1016/j.ultrasmedbio.2020.07.024DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7380221PMC
November 2020

Size-dependent anti-inflammatory activity of a peptide-gold nanoparticle hybrid in vitro and in a mouse model of acute lung injury.

Acta Biomater 2019 02 28;85:203-217. Epub 2018 Dec 28.

Department of Pulmonary and Critical Care Medicine, Shanghai General Hospital, Shanghai Jiao Tong University Shool of Medicine, Shanghai 201620, China. Electronic address:

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a life-threatening condition of critically-ill patients, characterized by overwhelming inflammatory responses in the lung. Multiple lines of evidence suggest that the excessive activation of Toll-like receptor 4 (TLR4) plays an important role in this detrimental lung inflammation. Recently, we developed a unique class of peptide-gold nanoparticle (GNP) hybrids that act as potent nano-inhibitors of TLR4 signaling by modulating the process of endosomal acidification. In this study, we aimed to identify the key physiochemical factors that could further strengthen the anti-inflammatory activity of these nano-inhibitors, including the nanoparticle size, the density of peptides coating the nanoparticle surface, as well as the number of the effective amino acid phenylalanine (F) residues in the peptide sequence. Among these factors, we found that the nanoparticle size could significantly affect the TLR4 inhibition. Specifically, the peptide-GNP hybrids with a GNP core of 20 nm (P12(G20)) exhibited the most potent inhibitory activity on TLR4 activation and its downstream cytokine production among those with a GNP core of 13 nm (P12(G13)) and 5 nm (P12(G5)) in THP-1 cell-derived macrophages. This size-dependent anti-inflammatory effect of the hybrid P12 was also observed in a lipopolysaccharide (LPS)-induced mouse model of ALI. It was found that P12(G20) was superior to P12(G13) in prolonging the survival of mice experiencing lethal LPS challenge, decreasing the acute lung inflammation, and alleviating diffuse alveolar damage in the lungs. Interestingly, P12(G20) could also promote long-term tolerance to endotoxin. Detailed mechanistic studies demonstrated that when compared to the smaller P12(G13), the larger P12(G20) had higher cellular uptake and a stronger endosomal pH buffering capacity, contributing to its enhanced therapeutic effects on reducing TLR4 activation in vitro and in vivo. Overall, this study suggests that nanoparticle size is one key factor determining the anti-inflammatory potency of the peptide-GNP hybrids, and the hybrid P12 may serve as a promising, novel class of nanotherapeutics for modulating TLR signaling to treat ALI/ARDS. STATEMENT OF SIGNIFICANCE: We have developed a new class of nanoparticle-based inhibitors (i.e., peptide-GNP hybrids) targeting TLR4 signaling in macrophages. Through evidence-based engineering of the nanoparticle size, surface peptide ligand density and effective amino acid (phenylalanine, F) chain length, we identified a peptide-GNP hybrid, P12(G20), with enhanced anti-inflammatory activity. Specifically, P12(G20) was more potent in reducing inflammation in THP-1 cell-derived macrophages and in a LPS-induced ALI mouse model. More interestingly, P12(G20) facilitated long-term protection against lethal LPS challenge in vivo and induced endotoxin tolerance in vitro. We anticipate that these new hybrids would serve as the next generation anti-inflammatory nano-therapeutics for the treatment of ALI/ARDS or other acute and chronic inflammatory diseases.
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http://dx.doi.org/10.1016/j.actbio.2018.12.046DOI Listing
February 2019

Application value of CT and MRI in diagnosis of primary brain lymphoma.

Oncol Lett 2018 Jun 30;15(6):8500-8504. Epub 2018 Mar 30.

Department of Interventional Medicine, People's Hospital of Rizhao, Rizhao, Shandong 276800, P.R. China.

This study explored the correlation between computed tomography (CT) and magnetic resonance imaging (MRI) manifestations and pathological features of primary brain lymphoma to improve the diagnostic accuracy. A total of 230 patients with primary brain lymphoma admitted to People's Hospital of Rizhao from July, 2005 to December, 2016 were selected into the study and their clinical data were analyzed retrospectively. Among them, 87 patients were examined by CT, 74 patients by MRI, 69 patients by both MRI and CT. Features of MRI and CT scanning figures were observed with a focus on the density, number and margins of the lesions, and the diagnostic accuracy was analyzed. A total of 353 lesions were identified from 230 primary brain lymphoma patients, of which 224 were single lesions, and 129 were multiple lesions. Most lesions were on the upper curtain (81.3%, 187 cases) and 43 cases (18.7%) were on the lower curtain. Lesion signal of CT and MRI plain scan showed uniform state, and enhanced scan showed significantly enhanced signal. Diagnostic accuracy of CT was 82.8%, and sensitivity and specificity was 75.5 and 67.4%, respectively. Diagnostic accuracy of MRI was 83.8%, and sensitivity and specificity was 79.3 and 64.9%, respectively. Diagnostic accuracy of MRI combined with CT was 89.9%, and sensitivity and specificity was 86.3 and 75.8%, respectively. CT combined with MRI can provide better diagnosis for primary brain lymphoma compared with CT or MRI alone, but pathological test is still needed.
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http://dx.doi.org/10.3892/ol.2018.8404DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5950549PMC
June 2018

Effects of ozone repeated short exposures on the airway/lung inflammation, airway hyperresponsiveness and mucus production in a mouse model of ovalbumin-induced asthma.

Biomed Pharmacother 2018 May 27;101:293-303. Epub 2018 Feb 27.

Department of Respiratory Medicine, Shanghai General Hospital of Nanjing Medical University, Shanghai, China; Department of Respiratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai, China. Electronic address:

Objective: The purpose of this study is to explore the influence of ozone repeated short exposures on airway/lung inflammation, airway hyperresponsiveness (AHR) and airway hypersecretion in ovalbumin (OVA) sensitized/challenged asthmatic mouse model.

Methods: OVA sensitization was performing by intraperitoneal injection. Ozone exposures (3ppm for 3hours) were given one hour after aerosolized OVA challenges (once every other day, 4 times totally). Methacholine (MCH) bronchial provocation tests, Liu's staining of BALF cell smears, hematoxylin-eosin (HE) staining and Periodic Acid-Schiff (PAS) staining of lung tissue were performed. Interleukins (ILs; IL-4, IL-13, IL-1β, and IL-18) protein (ELISA) and mRNA expression levels (RT-qPCR) in murine lung, 8-hydroxy-2'-deoxyguanosine (8-OHdG, ELISA), malondialdehyde (MDA, thiobarbituric acid assay), reduced glutathione (GSH, spectrophotometric method) in bronchoalveolar lavage fluid (BALF), and GSH1 mRNA relative expression levels (RT-qPCR) in lung tissue were analyzed.

Result: Repeated ozone exposures down-regulated the AHR to MCH in mice undergoing OVA sensitization and challenge, however not all parameters associated with asthma were decreased since obvious mucus hypersecretion was induced and airway inflammation increased slightly, especially around small airways. Following ozone co-exposure, the increase of IL-4 and IL-13 levels in murine lung caused by OVA sensitization/challenge were reversed. Instead, levels of IL-1β in BALF remained, higher than negative control group. Ozone repeated short exposures also induced significant increase of 8-OHdG in BALF in OVA sensitized and challenged mice.

Conclusion: For asthmatic mice undergoing ozone exposures, AHR is not an accurate indicator of the severity of asthma. Repeated short ozone exposures increase mucus hypersecretion, possibly via an increase in oxidative stress and immune dysregulation.
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http://dx.doi.org/10.1016/j.biopha.2018.02.079DOI Listing
May 2018

Involvements of p38 MAPK and oxidative stress in the ozone-induced enhancement of AHR and pulmonary inflammation in an allergic asthma model.

Respir Res 2017 12 29;18(1):216. Epub 2017 Dec 29.

Department of Respiratory Medicine, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, 100 Haining Road, Shanghai, 200080, People's Republic of China.

Background: Exposure to ambient ozone (O) increases the susceptivity to allergens and triggers exacerbations in patients with asthma. However, the detailed mechanisms of action for O to trigger asthma exacerbations are still unclear.

Methods: An ovalbumin (OVA)-established asthmatic mouse model was selected to expose to filtered air (OVA-model) or 1.0 ppm O (OVA-O model) during the process of OVA challenge. Next, the possible involvements of p38 MAPK and oxidative stress in the ozone actions on the asthma exacerbations were investigated on the mice of OVA-O model by treating them with SB239063 (a p38 MAPK inhibitor), and/or the α-tocopherol (antioxidant). Biological measurements were conducted including airway hyperresponsiveness (AHR), airway resistance (Raw), lung compliance (CL), inflammation in the airway lumen and lung parenchyma, the phosphorylation of p38 MAPK and heat shock protein (HSP) 27 in the tracheal tissues, and the malondialdehyde (MDA) content and the glutathione peroxidase (GSH-Px) activity in lung tissues.

Results: In OVA-allergic mice, O3 exposure deteriorated airway hyperresponsiveness (AHR), airway resistance (Raw), lung compliance (CL) and pulmonary inflammation, accompanied by the increased oxidative stress in lung tissues and promoted p38 MAPK and HSP27 phosphorylation in tracheal tissues. Administration of SB239063 (a p38 MAPK inhibitor) on OVA-O3 model exclusively mitigated the Raw, the CL, and the BAL IL-13 content, while α-tocopherol (antioxidant) differentially reduced the BAL number of eosinophils and macrophages, the content of BAL hyaluronan, the peribronchial inflammation, as well as the mRNA expression of TNF-α and IL-5 in the lung tissues of OVA-O3 model. Administration of these two chemical inhibitors similarly inhibited the AHR, the BAL IFN-γ and IL-6 production, the perivascular lung inflammation and the lung IL-17 mRNA expression of OVA-O3 model. Interestingly, the combined treatment of both compounds together synergistically inhibited neutrophil counts in the BALF and CXCL-1 gene expression in the lung.

Conclusions: O exposure during the OVA challenge process promoted exacerbation in asthma. Both p38 MAPK and oxidative stress were found to play a critical role in this process and simultaneous inhibition of these two pathways significantly reduced the O-elicited detrimental effects on the asthma exacerbation.
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http://dx.doi.org/10.1186/s12931-017-0697-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5747109PMC
December 2017

Recombinant human IL-26 facilitates the innate immune response to endotoxin in the bronchoalveolar space of mice in vivo.

PLoS One 2017 5;12(12):e0188909. Epub 2017 Dec 5.

Unit for Lung and Airway Research, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.

Interleukin (IL)-26 is released in response to bacterial endotoxin (LPS) in the bronchoalveolar space of humans in vivo and it may potentiate neutrophil chemotaxis by enhanced IL-26 receptor stimulation. However, the effects of extracellular IL-26 protein on the innate immune response in the lungs in vivo remain unknown. Here, we characterized these effects of IL-26 on a wide range of aspects of the innate immune response to LPS in different compartments of the lungs in vivo over time. We administrated recombinant human (rh) IL-26 protein in the bronchoalveolar space using intranasal instillation in a mouse in vivo model, with and without prior instillation of LPS. We verified gene expression of the IL-26 receptor complex in mouse lungs and observed that, after instillation of LPS, rhIL-26 increases the phosphorylation of STAT3, a signaling molecule of the IL-26 receptor complex. We also observed that rhIL-26 exerted additional stimulatory and inhibitory actions that are compartment- and time-dependent, resulting in alterations of cytokines, proteinases, tissue inflammation and the accumulation of innate effector cells. Without the prior instillation of LPS, rhIL-26 exerted time-dependent effects on total gelatinase activity but few other effects. Most important, after instillation of LPS, rhIL-26 cleared inflammatory cells from local tissue and increased the accumulation of innate effector cells in the bronchoalveolar space. Tentatively, rhIL-26 may facilitate the innate immune response towards the bronchoalveolar space in vivo and represents a potential target for therapy in lung disorders involving the innate immune response.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0188909PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716532PMC
December 2017

Screening Bioactive Nanoparticles in Phagocytic Immune Cells for Inhibitors of Toll-like Receptor Signaling.

J Vis Exp 2017 07 26(125). Epub 2017 Jul 26.

Department of Pediatrics, BC Children's Hospital and University of British Columbia.

Pharmacological regulation of Toll-like receptor (TLR) responses holds great promise in the treatment of many inflammatory diseases. However, there have been limited compounds available so far to attenuate TLR signaling and there have been no clinically approved TLR inhibitors (except the anti-malarial drug hydroxychloroquine) in clinical use. In light of rapid advances in nanotechnology, manipulation of immune responsiveness using nano-devices may provide a new strategy to treat these diseases. Herein, we present a high throughput screening method for quickly identifying novel bioactive nanoparticles that inhibit TLR signaling in phagocytic immune cells. This screening platform is built on THP-1 cell-based reporter cells with colorimetric and luciferase assays. The reporter cells are engineered from the human THP-1 monocytic cell line by stable integration of two inducible reporter constructs. One expresses a secreted embryonic alkaline phosphatase (SEAP) gene under the control of a promoter inducible by the transcription factors NF-κB and AP-1, and the other expresses a secreted luciferase reporter gene under the control of promoters inducible by interferon regulatory factors (IRFs).Upon TLR stimulation, the reporter cells activate transcription factors and subsequently produce SEAP and/or luciferase, which can be detected using their corresponding substrate reagents. Using a library of peptide-gold nanoparticle (GNP) hybrids established in our previous studies as an example, we identified one peptide-GNP hybrid that could effectively inhibit the two arms of TLR4 signaling cascade triggered by its prototypical ligand, lipopolysaccharide (LPS). The findings were validated by standard biochemical techniques including immunoblotting. Further analysis established that this lead hybrid had a broad inhibitory spectrum, acting on multiple TLR pathways, including TLR2, 3, 4, and 5. This experimental approach allows a rapid assessment of whether a nanoparticle (or other therapeutic compounds) can modulate specific TLR signaling in phagocytic immune cells.
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http://dx.doi.org/10.3791/56075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5612600PMC
July 2017

MiR-142-3p Overexpression Increases Chemo-Sensitivity of NSCLC by Inhibiting HMGB1-Mediated Autophagy.

Cell Physiol Biochem 2017 16;41(4):1370-1382. Epub 2017 Mar 16.

Department of Respiratory, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Background: Non-small-cell lung cancer (NSCLC) is a deadly cancer with high mortality rate. Drug resistance represents a main obstacle in NSCLC treatment. High mobility group box-1 (HMGB1) protein promotes drug resistance in NSCLC cells by activating protective autophagy.

Methods: In the current study, we investigated the regulatory role of microRNA-142-3p (miR-142-3p) in HMGB1-mediated autophagy of NSCLC cells and its impact on drug resistance of NSCLC in vitro and in vivo. HMGB1 was identified as a putative target gene of miR-142-3p by in silico analysis. Our luciferase reporter assay results confirmed that miR-142-3p directly targets the 3'-UTR of HMGB1 in NSCLC cells.

Results: MiR-142-3p overexpression suppressed while miR-142-3p knockdown increased HMGB1 mRNA and protein expression. Starvation induced HMGB1 expression and activated autophagy in NSCLC cells. The starvation-induced autophagy was inhibited by miR-142-3p overexpression or HMGB1 knockdown. Moreover, miR-142-3p overexpression or HMGB1 knockdown increased PI3K, Akt, and mTOR phosphorylation. Inhibition of PI3K or mTOR restored starvation-induced autophagy inhibited by miR-142-3p overexpression or HMGB1 knockdown.

Conclusions: These results demonstrated that miR-142-3p regulates starvation-induced autophagy of NSCLC cells by directly downregulating HMGB1 and subsequently activating the PI3K/Akt/mTOR pathway. Further, miR-142-3p overexpression inhibited anticancer drug-induced autophagy and increased chemo-sensitivity of NSCLC in vitro and in vivo. These findings shed light on the therapeutic potential of miR-142-3p in combating acquired NSCLC chemo-resistance.
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http://dx.doi.org/10.1159/000467896DOI Listing
June 2017

Autophagy is a regulator of TRAIL-induced apoptosis in NSCLC A549 cells.

J Cell Commun Signal 2017 Sep 18;11(3):219-226. Epub 2017 Jan 18.

Department of Respiratory, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai, 200080, China.

Autophagy, a catabolic process by which cytoplasmic components are degraded in lysosomes, plays an important role in the maintenance of cellular homeostasis. Dysregulation of autophagy is associated with several diseases. However, few studies have addressed the role of autophagy in the lung, and its role in lung diseases remains unclear. In the present study, we examined the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on autophagy in A549 cells and explored the underlying mechanisms. We showed that TRAIL promoted autophagosome formation, as detected by the levels of LC3-II, and its effect on promoting autophagy was dependent on the expression of the autophagy related genes (ATGs) Atg5, Atg7, and beclin-1. TRAIL-induced ATG expression was attenuated by JNK silencing or treatment with the JNK inhibitor SP600125, indicating the involvement of the JNK pathway. Crosstalk between autophagy and apoptosis was demonstrated by silencing the autophagy related genes Atg5, Atg7, and beclin-1, and the dependence of TRAIL-induced apoptosis on autophagy-related gene expression. Taken together, our results indicate that TRAIL promotes autophagy in A549 cells via a mechanism involving the modulation of ATG expression through the JNK pathway. Inhibition of autophagy enhanced TRAIL-induced cell proliferative inhibition and apoptosis in A549 cells.
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http://dx.doi.org/10.1007/s12079-016-0364-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5559391PMC
September 2017

Hydrogen Sulfide Prevents and Partially Reverses Ozone-Induced Features of Lung Inflammation and Emphysema in Mice.

Am J Respir Cell Mol Biol 2016 07;55(1):72-81

1 Department of Respiratory Medicine and.

Hydrogen sulfide (H2S), a novel signaling gasotransmitter in the respiratory system, may have antiinflammatory properties in the lung. We examined the preventive and therapeutic effects of H2S on ozone-induced features of lung inflammation and emphysema. C57/BL6 mice were exposed to ozone or filtered air over 6 weeks. Sodium hydrogen sulfide (NaHS), an H2S donor, was administered to the mice either before ozone exposure (preventive effect) or after completion of 6 weeks of ozone exposure (therapeutic effect). The ozone-exposed mice developed emphysema, measured by micro-computed tomography and histology, airflow limitation, measured by the forced maneuver system, and increased lung inflammation with augmented IL-1β, IL-18, and matrix metalloproteinase-9 (MMP-9) gene expression. Ozone-induced changes were associated with increased Nod-like receptor pyrin domain containing 3 (NLRP3)-caspase-1 activation and p38 mitogen-activated protein kinase phosphorylation and decreased Akt phosphorylation. NaHS both prevented and reversed lung inflammation and emphysematous changes in alveolar space. In contrast, NaHS prevented, but did not reverse, ozone-induced airflow limitation and bronchial structural remodeling. In conclusion, NaHS administration prevented and partially reversed ozone-induced features of lung inflammation and emphysema via regulation of the NLRP3-caspase-1, p38 mitogen-activated protein kinase, and Akt pathways.
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http://dx.doi.org/10.1165/rcmb.2015-0014OCDOI Listing
July 2016

[Effects of ozone oxidative stress on the airway hyperresponsiveness and mucus production in mice with acute allergic airway inflammation].

Zhonghua Jie He He Hu Xi Za Zhi 2015 Mar;38(3):179-84

Objective: To explore the impact of ozone on the airway hyperresponsinveness (AHR), airway inflammation and mucus production in an allergic asthma mouse model.

Methods: Twenty-eight female BALB/c mice were randomly divided into 4 equal groups: healthy control, ozone control, asthma model, and ozone intervention. For asthma model establishing, the mice were sensitized and challenged with ovalbumin, while the controls received saline. For ozone exposure, the mice were exposed to 2.0 ppm ozone for 3 hrs, while the control treatment group exposed to filtered air for 3 hrs. Some measurements were performed 24 hrs after the exposure, including AHR, pulmonary inflammation, mucus secretion, epithelial barrier function, and the level of oxidant stress.

Results: Compared with the asthma model group, mice in the ozone intervention group exhibited lower LogPC100Penh (0.22 ± 0.09 vs 0.50 ± 0.19, t = 3.06, P = 0.006), higher bronchoalveolar lavage (BAL) neutrophil numbers [(0.80 ± 0.21) x 10(3)/L vs (0.15 ± 0.06) x 10(3)/L, t = 3.63, P = 0.019] and BAL concentration of lower molecular weight hyaluronan (LMW-HA) [(111 ± 17) µg/L vs (35 ± 18) µg/L, t = 5.12 P = 0.000], TNF-α[(155 ± 30) µg/L vs (86 ±19) µg/L, t = 2.15, P = 0.044] and IL-13 [(65 ± 11) µg/L vs (33 ± 20) µg/L, t = 2.95, P = 0.008]. Mice in the ozone intervention group showed higher lung pathological inflammation score (2. 80 0.10 vs 1.92 ± 0.23, t =3.91, P = 0.000) and upregulated expressions of TNF-α mRNA (7.0 ± 1.5 vs 3.57 ± 1.20, t = 2.65, P = 0.014), CXCL-1 mRNA (7.0 ± 1.1 vs 2.5 ± 1.0, t = 4.12, P = 0.000) and IL-17 mRNA (28.8 ± 5.2 vs 16.4 ± 4.4, t = 6.33, P = 0.000). Ozone exposure on the asthmatic mice also caused higher percentage of PAS positive-staining epithelial cells [(76.2 ± 8.7) % vs (55.8 ± 14.4) %, t = 8.14, P = 0.000] and higher epithelial surface mucus volume [(721 ± 584) nl/mm2 vs (272 ± 185) nl/mm2, t = 5.78, P = 0.000] as well as the MUC5ac mRNA expression (15.4 ± 4.6 vs 7.0 ± 1.9, t = 4.37, P = 0.000). Besides, ozone exposure in the asthma model decreased epithelial cell density (82 ± 22 vs 116 ± 15, t = -10.1, P = 0.000), while increased the BAL concentration of albumin [(45 ± 6) g/L vs (33 ± 4) g/L, t = 3.89, P = 0.001] .

Conclusions: Ozone exaggerates AHR and pulmonary inflammation, and causes damages in epithelial cells and promotes the production of epithelial mucus.
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March 2015

Impact of ozone exposure on the response to glucocorticoid in a mouse model of asthma: involvements of p38 MAPK and MKP-1.

Respir Res 2014 Oct 8;15:126. Epub 2014 Oct 8.

Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiao tong University, 100 Haining Road, Shanghai 200080China.

Background: Molecular mechanisms involved in the oxidative stress induced glucocorticoids insensitivity remain elusive. The mitogen-activated protein kinase phosphatase (MKP) 1 mediates a part of glucocorticoids action and can be modified by exogenous oxidants. Whether oxidant ozone (O3) can affect the function of MKP-1 and hence blunt the response to corticotherapy is not clear.

Methods: Here we employed a murine model of asthma established with ovalbumin (OVA) sensitization and challenge to evaluate the influence of O3 on the inhibitory effect of dexamethasone on AHR and airway inflammation, and by administration of SB239063, a selective p38 MAPK inhibitor, to explore the underlying involvements of the activation of p38 MAPK and the expression of MKP-1.

Results: Ozone exposure not only aggravated the pulmonary inflammation and AHR, but also decreased the inhibitory effects of dexamethasone, accompanied by the elevated oxidative stress, airway neutrophilia, enhanced phosphorylation of p38 MAPK, and upregulated expression of IL-17. Administration of SB239063 caused significant inhibition of the p38 MAPK phosphorylation, alleviation of the airway neutrophilia, and decrement of the ozone-induced IL-17 expression, and partly restored the ozone-impaired effects of dexamethasone. Ozone exposure not only decreased the protein expression of MKP-1, but also diminished the dexamethasone-mediated induction process of MKP-1 mRNA and protein expression.

Conclusions: The glucocorticoids insensitivity elicited by ozone exposure on current asthma model may involve the enhanced phosphorylation of p38 MAPK and disturbed expression of MKP-1.
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http://dx.doi.org/10.1186/s12931-014-0126-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4196074PMC
October 2014

Activation of p38 mitogen-activated protein kinase in ovalbumin and ozone-induced mouse model of asthma.

Respirology 2013 Nov;18 Suppl 3:20-9

Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiaotong University, Shanghai, China.

Background And Objective: Ozone exposure worsens the development of allergen-induced asthma. The p38 mitogen-activated protein kinase (MAPK) pathway plays an important role in the development of the inflammatory response, airway hyperresponsiveness (AHR) and airway remodelling. In this study, the role of the p38 MAPK pathway on the effects of chronic ozone exposure in ovalbumin (OVA)-sensitized and -challenged mice was investigated.

Methods: Mice were sensitized and challenged with OVA followed by ozone exposure. Dexamethasone (Dex) and SB239063, a p38 MAPK inhibitor, were used as preventive treatment.

Results: Compared with OVA-challenged mice, ozone exposure of OVA-challenged mice led to enhanced recruitment of inflammatory cells in bronchoalveolar lavage fluid, increases in inflammation scores, collagen accumulation, bronchial wall thickness and messenger RNA levels of inflammatory cytokines, along with activation of p38 MAPK/HSP27 and downregulation of MAPK phosphatase-1 (MKP-1) in the lung tissue. Dex treatment partially attenuated lung inflammation, while the cotreatment of Dex and SB239063 effectively reduced lung inflammation, inhibited airway remodelling, inactivated p38 MAPK/HSP27 and upregulated MKP-1 in the lung tissue.

Conclusions: Ozone exposure aggravated airway inflammation, airway remodelling, activation of p38 MAPK and downregulation of MKP-1 in OVA-sensitized and -challenged mice, which was ineffectively controlled by corticosteroids. p38 MAPK activation is a likely pathway involved in corticosteroid insensitivity.
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http://dx.doi.org/10.1111/resp.12189DOI Listing
November 2013

Effects of acute ozone exposure on lung peak allergic inflammation of mice.

Front Biosci (Landmark Ed) 2013 Jun 1;18:838-51. Epub 2013 Jun 1.

Department of Respiratory Medicine, The Affiliated First People's Hospital of Shanghai Jiaotong University, Shanghai 200080, China.

Asthma exacerbations are often triggered by air pollution, including O3, whereas how patients with asthma exacerbations react to high levels of ambient ozone remain unknown. Here, we investigated the manner in which acute ozone exposure affects the pathophysiological characteristics of an asthma model on the premise of culminated allergic airway inflammation. The asthma model was constructed in mice, and enhanced pause (Penh), total and differential cell number, soluble mediator concentration, histopathology, and Muc5ac mRNA expression in the mice were observed. The results showed that ozone could induce airway hyperresponsiveness (AHR) in controls and an additional enhancement of preexisting AHR in asthmatic mice. When exposed to ozone, the asthmatic mice expressed more neutrophils, TNF-α, IL-13, and hyaluronan in bronchoalveolar lavage than controls. The mice with asthma and the controls both showed decreased epithelial cell density in the proximal and distal airways. Ozone aggravated the increased mucus production and mucin gene expression in mice with asthma. These results show that subjects with asthma may react differently to the same high level of ambient ozone, especially for those with asthma exacerbations.
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http://dx.doi.org/10.2741/4147DOI Listing
June 2013