Genetically engineered distal airway stem cell transplantation protects mice from pulmonary infection.

Severe pulmonary infection is a major threat to human health accompanied by substantial medical costs, prolonged inpatient requirements, and high mortality rates. New antimicrobial therapeutic strategies are urgently required to address the emergence of antibiotic resistance and persistent bacterial infections. In this study, we show that the constitutive expression of a native antimicrobial peptide LL-37 in transgenic mice aids in clearing Pseudomonas aeruginosa (PAO1), a major pathogen of clinical pulmonary infection. Orthotopic transplantation of adult mouse distal airway stem cells (DASCs), genetically engineered to express LL-37, into injured mouse lung foci enabled large-scale incorporation of cells and long-term release of the host defense peptide, protecting the mice from bacterial pneumonia and hypoxemia. Further, correlates of DASCs in adult humans were isolated, expanded, and genetically engineered to demonstrate successful construction of an anti-infective artificial lung. Together, our stem cell-based gene delivery therapeutic platform proposes a new strategy for addressing recurrent pulmonary infections with future translational opportunities.

Overexpression of transcription factor EB regulates mitochondrial autophagy to protect lipopolysaccharide-induced acute lung injury.

BACKGROUND: Acute lung injury (ALI) is characterized by an acute inflammatory process, and oxidative stress in the lung tissue leads to a lack of effective therapeutics. This study aimed to identify whether the overexpression of transcription factor EB (TFEB) regulates mitophagy to protect against lipopolysaccharide (LPS)-induced ALI. METHODS: We detected the expression of inflammatory factors, cytochrome c (Cyt.c) and nicotinamide adenine dinucleotide phosphate (NADPH), and autophagy-related proteins and observed the changes in lung histopathology induced by ALI in rats and the changes in the cell ultrastructure of primary alveolar type II epithelial cells induced by changing the expression of TFEB in the context of ALI. RESULTS: The overexpression of TFEB could reduce the expression of proinflammatory factors, such as IL-1 and IL-6, and increase the expression of anti-inflammatory factors, such as IL-10, both in vitro and in vivo. In addition, the overexpression of TFEB could reduce the Cyt.c and NADPH levels both in vivo and in vitro. The overexpression of TFEB could upregulate the expression of autophagy-related proteins, such as lysosomal-associated membrane protein 1 (LAMP1), microtubule-associated protein light chain 3B (LC3B), and Beclin both in vivo and in vitro, and promote mitochondrial autophagy. The overexpression of TFEB significantly improved the histopathologic changes induced by LPS-induced ALI in rats. However, low TFEB expression produced the opposite results. CONCLUSION: TFEB overexpression can decrease inflammation and mitochondrial damage in the lung tissue and alveolar epithelial cells through regulating mitochondrial autophagy to protect against LPS-induced ALI. Therefore, TFEB is likely a potential therapeutic target in LPS-induced ALI.

Management of Persistent Air Leaks Using Endobronchial Autologous Blood Patch and Spigot Occlusion: A Multicentre Randomized Controlled Trial in China.

BACKGROUND: Optimal management of persistent air leaks (PALs) in patients with secondary spontaneous pneumothorax (SSP) remains controversial. OBJECTIVE: To evaluate the efficacy and safety of endobronchial autologous blood plus thrombin patch (ABP) and bronchial occlusion using silicone spigots (BOS) in patients with SSP accompanied by alveolar-pleural fistula (APF) and PALs. METHODS: This prospective multicentre randomized controlled trial compared chest tube-attached water-seal drainage (CTD), ABP, and BOS that were performed between February 2015 and June 2017 in one of six tertiary care hospitals in China. Patients diagnosed with APF experiencing PALs (despite 7 days of CTD) and inoperable patients were included. Outcome measures included success rate of pneumothorax resolution at the end of the observation period (further 14 days), duration of air leak stop, lung expansion, hospital stay, and complications. RESULTS: In total, 150 subjects were analysed in three groups (CTD, ABP, BOS) of 50 each. At 14 days, 60, 82, and 84% of CTD, ABP, and BOS subjects, respectively, experienced full resolution of pneumothorax (p = 0.008). All duration outcome measures were significantly better in the ABP and BOS groups than in the CTD group (p < 0.016 for all). The incidence of adverse events, including chest pain, cough, and fever, was not significantly different. All subjects in the ABP and BOS groups experienced temporary haemoptysis. Spigot displacement occurred in 8% of BOS subjects. CONCLUSION: ABP and BOS resulted in clinically meaningful outcomes, including higher success rate, duration of air leak stop, lung expansion, and hospital stay, with an acceptable safety profile.

Seawater inhalation induces acute lung injury via ROS generation and the endoplasmic reticulum stress pathway.

Seawater (SW) inhalation can induce acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). In the present study, SW induced apoptosis of rat alveolar epithelial cells and histopathological alterations to lung tissue. Furthermore, SW administration increased generation of reactive oxygen species (ROS), whereas pretreatment with the ROS scavenger, N­acetyl­L­cysteine (NAC), significantly decreased ROS generation, apoptosis and histopathological alterations. In addition, SW exposure upregulated the expression levels of glucose­regulated protein 78 (GRP78) and CCAAT/enhancer binding protein homologous protein (CHOP), which are critical proteins in the endoplasmic reticulum (ER) stress response, thus indicating that SW may activate ER stress. Conversely, blocking ER stress with 4­phenylbutyric acid (4­PBA) significantly improved SW­induced apoptosis and histopathological alterations, whereas an ER stress inducer, thapsigargin, had the opposite effect. Furthermore, blocking ROS with NAC inhibited SW­induced ER stress, as evidenced by the downregulation of GRP78, phosphorylated (p)­protein kinase R­like ER kinase (PERK), p­inositol­requiring kinase 1α (IRE1α), p­50 activating transcription factor 6α and CHOP. In addition, blocking ER stress with 4­PBA decreased ROS generation. In conclusion, the present study indicated that ROS and ER stress pathways, which are involved in alveolar epithelial cell apoptosis, are important in the pathogenesis of SW­induced ALI.

Clinical application of ultrasound bronchoscopy guided needle aspiration biopsy under general anesthesia / 中国内镜杂志

Objective To explore the application of transbronchial needle aspiration biopsy (EBUS-TBNA) combined with ultrasound bronchoscopy under the general anesthesia (intravenous anesthesia combined with laryngeal mask). Methods 103 patients with EBUS-TBNA from June 2015 to October 2016 were randomly divided into local anesthesia group and general anesthesia group. The anesthesia group was anesthetized with intravenous anesthesia combined with laryngeal mask, local anesthesia group with conventional tetracaine local anesthesia. The differences in the average number of puncture needles, the diagnosis rate, the operation time, the complications and the comfort of each group were analyzed retrospectively. Results The average number of puncture needle in each group was 5.6 times, and the local anesthesia group was 3.1 times. The diagnosis rate of general anesthesia group was 94.12% and local anesthesia group was 78.85%. There was significant difference between the two groups (χ2 = 5.11, P = 0.024). The average operation time in the general anesthesia group was (14.61 ± 1.52), the local anesthesia group was (32.52 ± 4.14), they had significant difference (t = 29.01, P = 0.000). The number of patients in bleeding and cough of the general anesthesia group were lower than those in the local anesthesia group (χ2 = 4.31, 103.00, P = 0.038, 0.000, respectively). Conclusions EBUS-TBNA under general anesthesia has high diagnostic rate, shortoperating time, painlessness, high comfort and reduce operational risk, worthy of promoting clinical application.

Clinical application of ultrasound bronchoscopy guided needle aspiration biopsy under general anesthesia / 中国内镜杂志

Objective To explore the application of transbronchial needle aspiration biopsy (EBUS-TBNA) combined with ultrasound bronchoscopy under the general anesthesia (intravenous anesthesia combined with laryngeal mask). Methods 103 patients with EBUS-TBNA from June 2015 to October 2016 were randomly divided into local anesthesia group and general anesthesia group. The anesthesia group was anesthetized with intravenous anesthesia combined with laryngeal mask, local anesthesia group with conventional tetracaine local anesthesia. The differences in the average number of puncture needles, the diagnosis rate, the operation time, the complications and the comfort of each group were analyzed retrospectively. Results The average number of puncture needle in each group was 5.6 times, and the local anesthesia group was 3.1 times. The diagnosis rate of general anesthesia group was 94.12% and local anesthesia group was 78.85%. There was significant difference between the two groups (χ2 = 5.11, P = 0.024). The average operation time in the general anesthesia group was (14.61 ± 1.52), the local anesthesia group was (32.52 ± 4.14), they had significant difference (t = 29.01, P = 0.000). The number of patients in bleeding and cough of the general anesthesia group were lower than those in the local anesthesia group (χ2 = 4.31, 103.00, P = 0.038, 0.000, respectively). Conclusions EBUS-TBNA under general anesthesia has high diagnostic rate, shortoperating time, painlessness, high comfort and reduce operational risk, worthy of promoting clinical application.

Effects of para-toluenesulfonamide intratumoral injection on non-small cell lung carcinoma with severe central airway obstruction: A multi-center, non-randomized, single-arm, open-label trial.

BACKGROUND: Severe malignant airway obstruction (SMAO) is a life-threatening form of non-small cell lung carcinoma (NSCLC). OBJECTIVES: To determine the efficacy and safety of para-toluenesulfonamide (PTS) intratumoral injection in NSCLC-SMAO. METHODS: Ninety patients with NSCLC-SAO received repeated courses of PTS intratumoral injection until tumor sizes had reduced by 50% or greater. Primary endpoint was objective alleviation rate, assessed by chest computed tomography (CT) and bronchoscopy, at day 7 and 30 following final dosing. Secondary endpoints included airway obstruction, spirometry, quality-of-life and survival time. RESULTS: In full-analysis set (N=88), using RECIST criteria, PTS treatment resulted in a significant objective alleviation rate [chest CT: 59.1% (95%CI: 48.1%-69.5%), bronchoscopy: 48.9% (95%CI: 38.1%-59.8%) at day 7; chest CT: 43.2% (95%CI: 32.7%-54.2%), bronchoscopy: 29.6% (95%CI: 20.3%-40.2%) at day 30]. There was a remarkable increase in FVC (mean difference: 0.35 liters, 95%CI: 0.16-0.53 liters), FEV1 (mean difference: 0.27 liters, 95%CI: 0.07-0.48 liters), Baseline Dyspnea Index (mean difference: 64.8%, 95%CI: 53.9-74.7%) and Functional Assessment of Cancer Therapy-Lung Cancer Subscale (mean difference: 6·9, 95%CI: 3.8-9.9) at day 7 post-treatment. We noted significantly reduced prevalence of atelectasis (by 42.9%) and Eastern Cooperative Oncology Group physical performance scale (mean difference: 7.2, 95%CI: 3.9-10.5). Median survival time was 394 days in full-analysis set and 460 days in per-protocol set. Adverse events were reported in 64.0% of subjects. Seven severe adverse events (7.9%) were reported, of which three led to death (drug-related in one case). CONCLUSION: PTS intratumoral injection is effective and well tolerated for palliative therapy of NSCLC-SMAO.

Novel bronchoscopic balloon dilation for patients with bronchostenosis caused by bronchial tuberculosis: a case report.

INTRODUCTION: Bronchoscopic balloon dilation is a common method in the treatment of bronchostenosis but it is not an effective treatment due to its short dilating time (3 minutes) and low pressure (<3atm). Until recently, the reported highest dilating pressure was ≤6atm; however, this is not enough pressure to dilate a bronchostenosis because of the resistance of the bronchus. We hypothesized that higher dilating pressure (up to 14atm) with longer dilating time (40 minutes) may make bronchoscopic balloon dilation treatment more effective according to the blood vessel dilating method. Therefore, we designed this new bronchoscopic balloon dilation method for treating bronchostenosis, particularly in cases caused by bronchial tuberculosis. CASE PRESENTATION: A 23-year-old Chinese woman presented with right middle segmental bronchostenosis caused by bronchial tuberculosis. She was informed of the surgical procedure and she provided informed consent. After taking anti-bronchial tuberculosis drugs for 2 months, she underwent our new bronchoscopic balloon dilation treatment (dilating time, 40 minutes; pressure, 14atm). After anti-bronchial tuberculosis treatment for 13 months, her intermediate bronchus was observed with videobronchoscopy again and no re-stenosis was seen. Furthermore, a computed tomography scan revealed that her right lower lobe and right middle lobe had reopened. No complications occurred in the patient. CONCLUSION: The novel high-handed videobronchoscopic balloon dilation method was safe and effective for treating this patient with bronchostenosis caused by bronchial tuberculosis.

Participation of autophagy in acute lung injury induced by seawater.

Seawater drowning can lead to acute lung injury (ALI). However, the molecular and cellular mechanisms underlying this phenomenon remain elusive. The overall aim of this study is to clarify the role of autophagy in seawater-induced ALI, by which we can further understand the molecular mechanism and develop new methods for prevention and treatment of seawater-induced ALI. In this study, electron microscopy, western blot analysis, and RT-PCR were used to detect autophagy in lung tissues. Moreover, arterial blood gas analysis, lung weight coefficient, TNF-α, IL-8 in bronchoalveolar fluid (BALF), histopathology were used to detect the lung injury of seawater exposure. An inhibitor of autophagy (3-Methyladenine, 3-MA) was injected intraperitoneally before seawater exposure to further explore the role of autophagy in ALI. Electron microscopy revealed increasing autophagosomes in alveolar epithelial cell in seawater group compared with the control. The transcription and expression levels (mRNA and protein levels) of the LC3 II significantly increased in lung tissue of seawater group compared with those in control group. Furthermore, the alterations of autophage were basically consistent with the changes in arterial blood gas, lung weight coefficient, TNF-α, IL-8 in BALF and morphologic findings. In addition, inhibition of autophagy by 3-MA partly ameliorated seawater-induced ALI, as indicated by reduced lung weight coefficient and TNF-α in BALF, as well as increased PaO2. In conclusion, seawater aspiration triggered autophagy, and autophagy may be a scathing factor responsible for ALI induced by seawater.

Therapeutic effects of sequential therapy by electric coagulation, cryotherapy and balloon dilation with an electronic video bronchoscope.

The aim of the current study was to retrospectively analyze clinical data concerning bronchostenosis or bronchial obstruction caused by endobronchial tuberculosis. Fifty-six cases were subjected to bronchoscopy and chest computed tomography to assess the prognosis of bronchostenosis and bronchial obstruction. Based on reliable and effective anti-pulmonary tuberculosis therapy, these conditions were treated sequentially by electric coagulation, cryotherapy and balloon dilation with an electronic video bronchoscope during outpatient consultation or inpatient hospitalization. Fifty-three subjects with bronchostenosis recovered to varying degrees, a recovery rate of 94.6%. Thirteen of the 15 cases with bronchial obstruction reopened (86.7%). The clinical symptoms of these cases appeared to be in remission. Bronchostenosis or bronchial obstruction resulting from endobronchial tuberculosis may be treated by electric coagulation, cryotherapy and balloon dilation with an electronic video bronchoscope.

Osthole improves acute lung injury in mice by up-regulating Nrf-2/thioredoxin 1.

Inhibiting reactive oxygen species (ROS) has been viewed as a therapeutic target for the treatment of acute lung injury (ALI). Osthole, an active component in Chinese herbal medicine, has drawn increasing attention because of its various pharmacological functions, including anti-inflammatory and anti-oxidative activities. The aim of the present study was to examine the effects of osthole on ALI induced by lipopolysaccharide (LPS) through intratracheal instillation. The mRNA and protein expression levels of thioredoxin 1 (Trx1) and the nuclear factor erythroid-2 related factor 2 (Nrf2) were detected by real-time PCR, reverse transcription PCR (RT-PCR) and Western blot, respectively. ROS production was measured by flow cytometry. Our results showed that osthole treatment improved the mice survival rates in the middle and high dosage groups, compared with the untreated LPS group. Moreover, osthole treatment significantly improved LPS-induced lung pathological damage, and it decreased the lung injury scores, lung wet/dry ratios and the total protein level in Bronchoalveolar lavage fluid (BALF). Osthole treatment dramatically reduced the H2O2, MDA and OH levels in the lung homogenates. LDH and ROS were markedly reduced in the osthole+LPS group in vitro. Furthermore, osthole increased Nrf2 and Trx1 expression in terms of mRNA and protein in vivo and in vitro. Nrf2 siRNA (siNrf2) could suppress the beneficial effects of osthole on ALI. In conclusion, the current study demonstrates that osthole exerted protective effects on LPS-induced ALI by up-regulating the Nrf-2/Trx-1 pathway.

Antiinflammatory effects of matrine in LPS-induced acute lung injury in mice.

Matrine is one of the main active components of Chinese herb Sophora flavescens Ait (Kushen), which has been demonstrated to be effective in suppressing inflammation. The aim of the present study is to investigate the effect of matrine on LPS-induced lung injury. Lung injury was assessed by histological study and wet to dry weight ratios, as well as cell count and protein content in bronchoalveolar lavage fluid. We also detected MPO activity reflecting neutrophil infiltration and MDA activity examining oxidative stress in lung tissues. Cytokines and ROS production in cells were monitored by ELISA and flow cytometry, respectively. The results showed that high dose of matrine significantly reduced the mortality rate of mice with LPS administration. Treatment with matrine improved LPS-induced lung histopathologic changes, alleviated pulmonary edema and lung vascular leak, inhibited MPO and MDA activity,and reduced the production of inflammatory mediators including TNF-α, IL-6 and HMGB1. In vitro, matrine administration reduced the production of ROS and inflammatory factors, which was possibly associated with inhibition of NF-κB. In conclusion, the current study demonstrated that matrine exhibited a protective effect on LPS-induced acute lung injury by inhibiting of the inflammatory response, which may involve the suppression of ROS and tissue oxidative stress.

Tanshinone IIA attenuates seawater aspiration-induced lung injury by inhibiting macrophage migration inhibitory factor.

Inflammation takes responsibility for the seawater aspiration-induced lung injury. Tanshinone IIA (TIIA) can protect lipopolysaccharide-induced lung injury in mice through the inhibition of inflammation, but it is not reported whether TIIA have a protective effect on lung injury induced by seawater aspiration. Macrophage migration inhibitory factor (MIF) plays an important role in acute lung injury. In this study, we observed the effect of TIIA on the seawater aspiration-induced lung injury and the role of MIF in it. Seawater was aspirated into trachea of rats to make the lung injury model. TIIA was administered to investigate its beneficial effect on seawater-induced acute lung injury. The results showed that seawater aspiration led to hyoxemia, pulmonary edema, neutrophil infiltration, and lung histopathologic changes, with the elevated MIF expression in the lung tissues and plasma. However, these changes were attenuated by TIIA. In macrophage cells we also demonstrated that TIIA could inhibit MIF expression, nuclear factor κB (NF-κB) activity and release of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) induced by seawater. Besides, pretreatment with (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid (ISO-1), the MIF antagonist, elevated NF-κB and cytokines induced by seawater were also reduced markedly. Furthermore, rMIF treatment alone increased the phosphorylation level of NF-κB and release of cytokines, which was almost abolished by TIIA. Taken together, our results suggested that TIIA exert a protective effect on the seawater aspiration-induced lung injury partly through downregulation of MIF and the subsequent NF-κB activity, as well as expression of IL-6 and TNF-α.

Sodium tanshinone iia sulfonate attenuates seawater aspiration-induced acute pulmonary edema by up-regulating Na(+),K(+)-ATPase activity.

Relieving pulmonary edema is the key of a successful treatment to seawater drowning. Sodium tanshinone IIA sulfonate (STS) has been observed to reduce lung edema from lipopolysaccharide (LPS)-induced lung injury. In this study the authors investigated whether STS attenuates seawater aspiration-induced acute pulmonary edema, and examined the effects of sodium-potassium adensosine triphosphatase (Na(+),K(+)-ATPase) on it. Seawater was instilled through an endotracheal tube. The anesthetized and spontaneously breathing rats received STS intraperitoneally after seawater aspiration. Pao(2), lung wet-to-dry weight ratio, and pulmonary microvascular permeability were tested. The authors explored the effects of STS on the expression and activity of Na(+),K(+)-ATPase in vivo and in vitro. Additionally, the authors investigated the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway in the stimulation of Na(+),K(+)-ATPase by STS. The results showed that STS significantly improved hypoxemia, attenuated lung edema, and alleviated seawater-induced lung injury in vivo. Both in vivo and in vitro, it was observed that STS up-regulated the expression and activity of Na(+),K(+)-ATPase. ERK1/2 inhibitor partially blocked the effects of STS on Na(+),K(+)-ATPase activity in alveolar type II cells following seawater incubation. These results indicated that STS could improve seawater aspiration-induced acute pulmonary edema by up-regulating Na(+),K(+)-ATPase activity, and the ERK1/2 signaling pathway may be involved in it.

Tanshinone IIA suppresses lung injury and apoptosis, and modulates protein kinase B and extracellular signal-regulated protein kinase pathways in rats challenged with seawater exposure.

1. Tanshinone IIA (TIIA) is one of the main active components of the Chinese herb, Danshen. In the present study, we investigated the role of apoptosis in seawater exposure-induced acute lung injury (ALI), and explored the effects of TIIA on lung injury, apoptosis, and protein kinase B (Akt) and extracellular signal-regulated protein kinase (ERK) pathways in seawater-challenged rats. The rats were randomly divided into four groups: (i) naive group, no drug was given; (ii) TIIA control group, TIIA (50 mg/kg) was given intraperitoneally; (iii) seawater (SW) group, seawater (4 mL/kg) was given; and (iv) TIIA/SW group, TIIA (50 mg/kg) was injected intraperitoneally 10 min after seawater instillation. 2. The results showed that TIIA treatment significantly improved seawater exposure-induced lung histopathological changes, alleviated the decrease in PaO(2) , and reduced lung oedema, vascular leakage and cell infiltration. As shown by terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) assay, seawater exposure induced apoptosis in lung tissue cells. Furthermore, seawater exposure also changed apoptosis-related factors Bcl-2 and caspase-3, and caused a reduction in the activation of Akt and ERK1/2 pathways. Furthermore, TIIA treatment decreased the number of apoptotic cells, reversed changes in Bcl-2 and caspase-3, and upregulated the activation of Akt and ERK1/2 in seawater-challenged rats. 3. In conclusion, the data suggest that apoptosis might play an important role in seawater exposure-induced lung injury and that TIIA could significantly attenuate the severity of ALI and apoptosis in seawater-challenged rats, which is possibly through modulation of Akt and ERK1/2 pathways.

[Effect of sulfonic tanshinone sodium injection on the expression and activity of aquaporin-5 of human alveolar epithelial cells after seawater exposure].

OBJECTIVE: To explore the effects of tanshinone IIA on the activity of aquaporin-5 (AQP5) in human alveolar epithelial cells (A549) after seawater exposure and its possible mechanism. METHODS: Routinely cultured A549 cells were divided into different groups according to different content of seawater: blank control group, 15%, 25%, 50%, 75%, 100% seawater groups; they were divided into different groups according to the duration of exposure to 25% seawater: blank control group, 1, 4, 8 hours groups; they were also divided into different groups according to concentration of tanshinone IIA and exposed to seawater for 4 hours: blank control group, 25% seawater group, 25, 50, 75, 100 µg/ml tanshinone IIA intervention groups. The expressions of AQP5 were respectively assayed by Western blotting and immunohistochemistry. RESULTS: The results of Western blotting showed that the expressions of AQP5 were remarkably higher at 8 hours of exposure to seawater in 25% and 50% seawater groups than those in blank control group (1.053±0.231, 1.116±0.316 vs. 0.101±0.081, both P<0.05); the expression of AQP5 in 1-hour group showed a slight increase compared with blank control group (0.306±0.125 vs. 0.288±0.098, P>0.05), that in 4-hour group was increased significantly (1.423±0.377, P<0.01), and in 8-hour group (1.507±0.461) it was slightly higher than that in 4-hour group without statistical significance. The AQP5 expression was significantly lower in tanshinone IIA 25 µg/ml and 50 µg/ml intervention groups than that in 25% seawater group (0.580±0.186, 0.499±0.172 vs. 1.013±0.287, both P<0.05). Immuno-histochemistry showed that the expression of AQP5 was markedly up-regulated after A549 cells were stimulated with 25% seawater for 4 hours as compared with blank control group (7.21±0.78 vs. 0.41±0.07, P<0.01), but intervention of tanshinone IIA significantly inhibited the up-regulation of AQP5 expression (3.02±0.23) induced by 25% seawater (P<0.05). CONCLUSION: The experimental results showed that tanshinone IIA is innocuous to A549 at a dosage of 25 µg/ml, and it can decrease the overexpression of AQP5 induced by seawater.

Overexpression of STAT3 potentiates growth, survival, and radioresistance of non-small-cell lung cancer (NSCLC) cells.

OBJECTIVE: Activation of signal transducer and activator of transcription 3 (STAT3) play important roles in tumorigenesis and tumor progression. The overexpression of STAT3 has been found in various malignancies including non-small-cell lung carcinoma (NSCLC). The purpose of this study was to explore the correlation between overexpression of STAT3 gene and growth, survival, and radiosensitivity of NSCLC cells. METHODS: Subclones using vector-based short hairpin RNA (shRNA) were established. RT-PCR and Western blot assays were performed to detect the expression of STAT3 mRNA and protein in untransfected or stably transfected NSCLC cells. Then, MTT and soft agar colony assays were performed to determine the effect of STAT3 inhibition on in vitro growth of NSCLC cells. Hoechst staining assay was performed to analyze the effect of STAT3 inhibition on apoptosis of NSCLC cells. Additionally, clonogenic survival assays were performed to detect the effect of STAT3 inhibition on in vitro radiosensitivity of NSCLC cells. Finally, to examine the effect of pSUPER-shSTAT3 on proliferation and radiosensitivity in vivo, a subcutaneous (s.c.) tumor formation assay in nude mice was performed. RESULTS: We successfully established two stable transfected cell lines (A549/shSTAT3 and SK-MES-1/shSTAT3) in which the expression of STAT3 mRNA and protein was down-regulated. Those two stable subclones showed a significantly dramatic reduction in colony-forming ability and proliferation not only in vitro but also in vivo. The apoptotic rates of A549/shSTAT3 and SK-MES-1/shSTAT3 cells increased to 19.2% and 16.4%, respectively. Moreover, shRNA-mediated STAT3 inhibition could also significantly enhance radiosensitivity of NSCLC cells both in vitro and in vivo. CONCLUSION: Together, the overexpression of STAT3 is correlated with growth, survival, and radioresistance of NSCLC cells, and STAT3 might be a molecular therapeutic target for gene therapy or radiosensitization of NSCLC.

[The application of combined interventional procedures for the management of intractable central airway stenosis].

OBJECTIVE: to evaluate the application of combined five interventional procedures in the management of intractable central airway stenosis. METHODS: clinical manifestations and pulmonary functions of 138 patients with intractable central airway stenosis were evaluated. Five interventional procedures, including high-frequency electrotome, argon plasma coagulation (APC), cryotherapy, stent placement and high-pressure balloon dilatation, were used in this study. Among them, two or more procedures were combined to manage complicated airway stenosis according to stenosis causes, types, position, degree and duration, or functions of distal lung tissue and airway. Forty-two cases were treated with high-frequency electrotome and APC, 54 cases with high-frequency electrotome, cryotherapy and APC, 29 cases with high-frequency electrotome, cryotherapy, APC and stent placement, and 13 cases with cryotherapy, APC and high-pressure balloon dilatation. Airways opening, short-term therapeutic effects and improvement of pulmonary functions were evaluated when ideal curative effects were achieved in the first month after intervention. RESULTS: the total short-term effective rate in the 138 patients was 100%. The airway diameter was increased from (2.6 ± 1.5) mm before operation to (6.2 ± 1.7) mm after operation (P < 0.05). Dyspnea score was decreased from (2.4 ± 0.8) before operation to (0.7 ± 0.6) after operation (P < 0.05). FEV(1) was increased from (1.8 ± 0.6) L before operation to (3.1 ± 0.7) L after operation (P < 0.05). Among 23 cases benign disease, including 4 benign tumor, 15 tuberculosis and 4 other granulomatosis, 5 cases with various degrees of restenosis needed further interventional therapy after 3 months of follow up and effective rate was 78.3% (18/23). After 6 months of follow-up, 3 cases with restenosis needed re-intervention, and the effective rate was 86.9% (20/23). All of the 23 cases did not experience stenosis after 12 months of follow-up. Patients with malignant tumor were not followed up for long term. CONCLUSION: combination of five interventional procedures, including high-frequency electrotome, APC, cryotherapy, stent placement and high-pressure balloon dilatation, has fewer complications and favorable clinical effects in management of intractable central airway stenosis.

[Joint application of 7 interventional pulmonology methods in early diagnosis of lung cancer].

OBJECTIVE: To evaluate the combination of 7 interventional pulmonology methods in early diagnosis of lung cancer. METHODS: A total of 467 patients with thoracic and pulmonary lesions (include hilum pulmonis lymphadenectasis, mediastinal lymphadenectasis, pulmonary scobination, lump, lamellar infiltration, small amount of pleural fluid and pleural scobination) had negative results via exfoliative cytology, bacteriology and routine bronchoscopy. All these patients had ultrathin bronchoscopy with biopsy and brushing. For those 155 cases whose foci were located at porta pulmonis, inner zone or median zone, the authors applied ultrathin bronchoscopy with biopsy and brushing guided by X-ray. For those 95 cases whose foci were located at median zone or outer zone and unconnected with chest wall, per cutem lung puncture needle aspiration was employed under the guidance of X-ray. For those 102 cases whose foci were tightly connected with pleural membrane, per cutem lung puncture biopsy was employed under the guidance of type-B ultrasonic. For those 59 cases with suspected central airway foci, auto-fluorescence bronchoscopic biopsy and brushing were employed. For those 67 cases with hilum pulmonis or mediastinal lymphadenectasis, endobronchial ultrasonic transbronchial needle aspiration (EBUS-TBNA) was employed. For those 23 cases with small amount of pleural fluid or pleural scobination, electronic thoracoscopic biopsy and brushing were employed. RESULTS: It was found that 118 cases were diagnosed by ultrathin bronchoscopic biopsy and brushing with a positive rate of 25.3% (118/467), 105 cases by ultrathin bronchoscopy with biopsy and brushing guided by X-ray with a positive rate of 67.7% (105/155), 63 cases by per cutem lung puncture needle aspiration under the guidance of X-ray with a positive rate of 66.3% (63/95), 69 cases by per cutem lung puncture biopsy under the guidance of type-B ultrasound with a positive rate of 67.6% (69/102), 18 cases by auto-fluorescence bronchoscopic biopsy and brushing with a positive rate of 35.3% (18/51), 52 cases by EBUS-TBNA with a positive rate of 77.6% (52/67), 12 cases by electronic thoracoscopic biopsy and brushing with a positive rate of 52.2% (12/23). The total positive diagnostic rate was 93.6% (437/467). And the diagnostic rate of < or = stage II lung cancer (3 cases carcinoma in situ, 84 stage I a, 63 stage Ib, 65 stage IIa and 44 stage IIb) was 82.7% (259/313). CONCLUSION: Joint application of these 7 interventional bronchoscopic techniques can significantly boost the rate of early diagnosis of lung cancer.

[Efficacy and safety of argon plasma coagulation in the treatment of obstructive tracheobronchial stenosis].

OBJECTIVE: To evaluate the efficacy and safety of argon plasma coagulation (APC) in the treatment of large airway obstruction. METHODS: Totally 389 patients with treacheobronchial stenosis were treated with APC (ARCO3000 type) by bronchoscopy. The stenoses were caused by carcinomas (203 cases, 52.2%), metastatic tumors (67 cases, 17.2%), benign tumors (18 cases, 4.6%), granulomas (93 cases, 23.9%) and other lesions (8 cases, 2.1%). The rate of recanalization, relief of the symptoms, and complications were analyzed. RESULTS: 1121 times of APC treatment were performed in the 389 patients. Complete recanalization was achieved in 138 cases (35.5%), partial in 143 (36.8%), mild in 55 (14.1%) and none in 53 (13.6%). The major complications included: super-ventricular tachycardia in 136 cases (34.9%), bleeding in 51 (13.1%), decrease in blood oxygen saturation in 48 (12.3%), asphyxia in 33 (8.5%), ventricular or super-ventricular arrhythmia in 24 (6.2%), short-term aggravation of airway obstruction in 18 (4.6%), and tracheal perforation in 3 (0.78%). All those complications were treated with various measures and no patient died of the complications during the procedure. CONCLUSION: Argon plasma coagulation is effective and relatively safe in relieving the obstruction and dyspnea in patients with large airway obstruction caused by various reasons. However, for the patients with severe airway obstruction, argon plasma coagulation sometimes may cause severe or even lethal complications. Critical consideration of the indication, operators' skill and taking more precautions during the procedure are required to ensure the safety of argon plasma coagulation treatment.