Central nervous system efficacy of aumolertinib versus gefitinib in patients with untreated, EGFR-mutated, advanced non-small cell lung cancer: data from a randomized phase III trial (AENEAS).

BACKGROUND: The initial randomized, double-blinded, actively controlled, phase III ANEAS study (NCT03849768) demonstrated that aumolertinib showed superior efficacy relative to gefitinib as first-line therapy in epidermal growth factor receptor (EGFR)-mutated advanced non-small cell lung cancer (NSCLC). Metastatic disease in the central nervous system (CNS) remains a challenge in the management of NSCLC. This study aimed to compare the efficacy of aumolertinib versus gefitinib among patients with baseline CNS metastases in the ANEAS study. METHODS: Eligible patients were enrolled and randomly assigned in a 1:1 ratio to orally receive either aumolertinib or gefitinib in a double-blinded fashion. Patients with asymptomatic, stable CNS metastases were included. Follow-up imaging of the same modality as the initial CNS imaging was performed every 6 weeks for 15 months, then every 12 weeks. CNS response was assessed by a neuroradiological blinded, independent central review (neuroradiological-BICR). The primary endpoint for this subgroup analysis was CNS progression-free survival (PFS). RESULTS: Of the 429 patients enrolled and randomized in the ANEAS study, 106 patients were found to have CNS metastases (CNS Full Analysis Set, cFAS) at baseline by neuroradiological-BICR, and 60 of them had CNS target lesions (CNS Evaluable for Response, cEFR). Treatment with aumolertinib significantly prolonged median CNS PFS compared with gefitinib in both cFAS (29.0 vs. 8.3 months; hazard ratio [HR] = 0.31; 95% confidence interval [CI], 0.17-0.56; P < 0.001) and cEFR (29.0 vs. 8.3 months; HR = 0.26; 95% CI, 0.11-0.57; P < 0.001). The confirmed CNS overall response rate in cEFR was 85.7% and 75.0% in patients treated with aumolertinib and gefitinib, respectively. Competing risk analysis showed that the estimated probability of CNS progression without prior non-CNS progression or death was consistently lower with aumolertinib than with gefitinib in patients with and without CNS metastases at baseline. No new safety findings were observed. CONCLUSIONS: These results indicate a potential advantage of aumolertinib over gefitinib in terms of CNS PFS and the risk of CNS progression in patients with EGFR-mutated advanced NSCLC with baseline CNS metastases. TRIAL REGISTRATION: ClinicalTrials.gov number, NCT03849768.

AENEAS: A Randomized Phase III Trial of Aumolertinib Versus Gefitinib as First-Line Therapy for Locally Advanced or MetastaticNon-Small-Cell Lung Cancer With EGFR Exon 19 Deletion or L858R Mutations.

PURPOSE: Aumolertinib (formerly almonertinib; HS-10296) is a novel third-generation epidermal growth factor receptor tyrosine kinase inhibitor approved in China. This double-blind phase III trial evaluated the efficacy and safety of aumolertinib compared with gefitinib as a first-line treatment for locally advanced or metastatic EGFR-mutated non-small-cell lung cancer (NSCLC; ClinicalTrials.gov identifier: NCT03849768). METHODS: Patients at 53 sites in China were randomly assigned 1:1 to receive either aumolertinib (110 mg) or gefitinib (250 mg) once daily. The primary end point was progression-free survival (PFS) per investigator assessment. RESULTS: A total of 429 patients who were naïve to treatment for locally advanced or metastatic NSCLC were enrolled. PFS was significantly longer with aumolertinib compared with gefitinib (hazard ratio, 0.46; 95% CI, 0.36 to 0.60; P < .0001). The median PFS with aumolertinib was 19.3 months (95% CI, 17.8 to 20.8) versus 9.9 months with gefitinib (95% CI, 8.3 to 12.6). Objective response rate and disease control rate were similar in the aumolertinib and gefitinib groups (objective response rate, 73.8% and 72.1%, respectively; disease control rate, 93.0% and 96.7%, respectively). The median duration of response was 18.1 months (95% CI, 15.2 to not applicable) with aumolertinib versus 8.3 months (95% CI, 6.9 to 11.1) with gefitinib. Adverse events of grade ≥ 3 severity (any cause) were observed in 36.4% and 35.8% of patients in the aumolertinib and gefitinib groups, respectively. Rash and diarrhea (any grade) were observed in 23.4% and 16.4% of patients who received aumolertinib compared with 41.4% and 35.8% of those who received gefitinib, respectively. CONCLUSION: Aumolertinib is a well-tolerated third-generation epidermal growth factor receptor tyrosine kinase inhibitor that could serve as a treatment option for EGFR-mutant NSCLC in the first-line setting.

CircRNA_103762 promotes multidrug resistance in NSCLC by targeting DNA damage inducible transcript 3 (CHOP).

BACKGROUND: CircRNAs have been found to play crucial roles in multiple tumor including non-small cell lung cancer (NSCLC). Here, we researched the correlation between circRNA_103762 and chemotherapy resistance. METHODS: RT-PCR assay was performed to detect circRNA_103762 and DNA damage inducible transcript 3 (CHOP) expression. CCK8 assay was performed to examine cell proliferation and IC50 of different drug. Migration and invasion assay was used to detect cell migration and invasion. RESULTS: In our study, circRNA_103762 expression was upregulated in NSCLC tissues and cell. Knockdown of circRNA_103762 can inhibited cell proliferation, migration and invasion in NSCLC. In addition, downregulation of circRNA_103762 promoted CHOP expression and inhibited multidrug resistance (MDR) in NSCLC. CONCLUSION: Together, we demonstrated that circRNA_103762 is upregulated in NSCLC and functions as an oncogene in NSCLC, and circRNA_103762 enhanced MDR by inhibited CHOP expression in NSCLC cells. These results will help us understand the MDR of NSCLC, providing better effective therapy strategies for patients.

1,25-Dihydroxyvitamin D3 prevents toluene diisocyanate-induced airway epithelial barrier disruption.

The loss of airway epithelial integrity contributes significantly to asthma pathogenesis. Evidence suggests that vitamin D plays an important role in the prevention and treatment of asthma. However, its role in airway epithelial barrier function remains uncertain. We have previously demonstrated impaired epithelial junctions in a model of toluene diisocyanate (TDI)-induced asthma. In the present study, we hypothesized that 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] may prevent TDI-induced epithelial barrier disruption. Male BALB/c mice were dermally sensitized and then challenged with TDI. The mice were then administered 1,25(OH)2D3 intraperitoneally prior to challenge with TDI. For in vitro experiments, 16HBE bronchial epithelial cells were cultured and stimulated with TDI-human serum albumin (HSA). The results revealed that the mice treated with 1,25(OH)2D3 displayed decreased airway hyperresponsiveness (AHR), suppressed neutrophil and eosinophil infiltration into the airways, as well as an increased E-cadherin and zonula occludens-1 (ZO-1) expression at the cell-cell contact sites. In vitro, exposure of the cells to TDI-HSA induced a rapid decline in transepithelial electrical resistance (TER) and an increase in cell permeability, followed by a decrease in occludin expression and the redistribution of E-cadherin, accompanied by a significant upregulation in the levels of phosphorylated extracellular signal-regulated kinase (ERK)1/2. These effects were all partly reversed by treatment with either 1,25(OH)2D3 or an ERK1/2 inhibitor. In conclusion, the findings of our study demonstrate that 1,25(OH)2D3 prevents TDI-induced epithelial barrier disruption, and that the ERK1/2 pathway may play a role in this process.

Invasive pulmonary aspergillosis in non-neutropenic patients with and without underlying disease: a single-centre retrospective analysis of 52 subjects.

BACKGROUND AND OBJECTIVE: Invasive pulmonary aspergillosis (IPA) remains a life-threatening infection in patients with prolonged neutropenia. Few data are available on IPA in non-neutropenic patients without underlying immunocompromising conditions. METHODS: All non-neutropenic patients managed at the institution for a proven and probable IPA over the last 10 years were reviewed retrospectively, and the difference between non-neutropenic patients with and without underlying disease was investigated. RESULTS: Among 52 cases of IPA analysed here, 33 were histologically proven; 19 were probable. Forty-two (80.8%) patients had underlying diseases; 10 (19.2%) patients had no any underlying diseases. There is a significant difference in seasonal distribution among patients with underlying conditions (P = 0.026), but no seasonal difference was found in the other group (P = 0.622). The only significant difference in symptoms between the two groups was fever (P = 0.015). Radiological findings were non-specific in the two groups. Despite treatment, the overall crude mortality rate among 52 patients was 39%. The overall mortality rate in patients with underlying disease was 45%, while that in patients without underlying conditions was 11%. A Cox multivariate analysis showed that organ failure (hazard ratios: 8.739, 95% CI: 3.770-20.255; P = 0.000) was independently associated with overall mortality. CONCLUSIONS: Clinical features of IPA are not well known in non-neutropenic patients, especially in those without underlying conditions. In this study, organ failure was associated with a lower rate of survival of non-neutropenic patients with IPA.

[Role of the AdeABC efflux pump in carbapenems resistance of clinical isolates of Acinetobacter baumannii].

OBJECTIVE: To investigate the role of AdeABC efflux pump in carbapenems resistance of Acinetobacter baumannii in light of the phenotype and genetype of the efflux pump. METHODS: The phenotype of the efflux pump was detected in 138 clinical isolates of A.baumannii using the efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone (CCCP). The mRNA expression of pump-encoding gene adeB in the strains was detected using quantitative real-time RT-PCR. RESULTS: Of the 138 strains, 28 showed positivities for AdeABC efflux pump identified by Mueller-Hinton Broth with CCCP. Of the 39 strains resistant to meropenem, 15 (38.4%) showed positive results in CCCP assay, a rate significantly higher than that among the 99 sensitive strains (13.1%, 13/99) (X(2)=12.477(b), P=0.01). The mRNA expression of efflux pump-encoding gene adeB was detected by real-time RT-PCR at a level of 0.899∓∓1.172 in meropenem-sensitive strains, significantly lower than the level of 21.101∓∓21.443 in meropenem-resistant strains (t=4.403, P=0.000). CONCLUSIONS: Efflux plays a role in carbapenems resistance in the clinical isolates of A. baumannii. The AdeABC efflux pump may be an important factor in reducing carbapenems sensitivity in A. baumannii.

[The comparison between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice on lung tissue pathological change and vasopermeability in acute lung injury].

OBJECTIVE: To compare the change of lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice in acute lung injury. METHODS: The mice with vascular endothelial cell-specific expression of cre recombinase were crossed with cdc42(flox/flox) mice. The cdc42(flox/+)Cre(+/-) F1 offspring mice were crossed back with cdc42(flox/flox) mice, resulting in the F2 generation mice with three genotypes, namely cdc42(flox/+)Cre(+/-), cdc42(flox/flox)Cre(-/-) and cdc42(flox/+)Cre(+/-). The heterozygous mice with cdc42(flox/+)Cre(+/-) genotype were selected as the model mice, with the other two genotype groups as the control. After intratracheal instillation of 2 mg/kg LPS to induce acute lung injury, the mice were sacrificed to examine the lung pathologies, lung wet/dry ratio and lung microvascular permeability. RESULTS: The heterozygous mice with cdc42 gene knockout (cdc42(flox/+)Cre(+/-)) showed no significant differences from the two control groups in the lung pathological score, lung wet/dry ratio or the lung microvascular permeability coefficient. CONCLUSION: There were no significant difference on lung tissue and vasopermeability between the vascular endothelial cells special cdc42-deficient heterozygous mice and the non-knockout mice.

Increased heat shock protein 70 levels in induced sputum and plasma correlate with severity of asthma patients.

Damage-associated molecular pattern molecules such as high-mobility group box 1 protein (HMGB1) and heat shock protein 70 (HSP70) have been implicated in the pathogenesis of asthma. The aim of our study was to examine the induced sputum and plasma concentrations of HSP70 in asthmatic patients to determine their relationship with airway obstruction. Thirty-four healthy controls and 56 patients with persistent bronchial asthma matched for gender and age were enrolled in this study. Spirometry measurements were performed before sputum induction. HSP70 levels in induced sputum and plasma were measured using the ELISA Kit. Sputum and plasma concentrations of HSP70 in asthmatics patients were significantly higher than that in control subjects (sputum, (0.88 ng/ml (0.27-1.88 ng/ml) versus 0.42 ng/ml (0.18-0.85 ng/ml), p < 0.001); plasma, (0.46 ng/ml (0.20-0.98 ng/ml) versus 0.14 ng/ml (0.11-0.37 ng/ml), p < 0.001) and were significantly negatively correlated with forced expiratory volume in 1 s (FEV1), FEV1 (percent predicted), and FEV1/FVC in all 90 participants and 56 patients with asthma. There were no significant differences in HSP70 levels between patients with eosinophilic and non-eosinophilic asthma. HSP70 levels in plasma were positively correlated with neutrophil count, and HSP70 levels in induced sputum were positively correlated with lymphocyte count. In multivariate analysis, independent predictors of sputum HSP70 were diseases and disease severity but not smoking, age, or gender, and independent predictors of plasma HSP70 were also diseases and disease severity. In conclusion, this study indicates that induced sputum and plasma HSP70 could serve as a useful marker for assessing the degree of airway obstruction in patients with asthma. However, further investigation is needed to establish the role of circulating and sputum HSP70 in the pathogenesis of asthma.

[Production of connective tissue growth factor by angiotensin II in human embryonic lung fibroblast via RhoA-ROCK pathway].

OBJECTIVE: To explore the production of connective tissue growth factor (CTGF) by Angiotensin II (AngII) in human embryonic lung fibroblast via the RhoA-ROCK pathway. METHODS: Human embryonic lung fibroblast (HFL-1) was divided into 4 groups: (1) control group: no stimulation; (2) AngII group: stimulation of AngII (10(-7) mol/L) ; (3) Irbesartan plus AngII group: stimulation by AngII (10(-7) mol/L) with AT-1 receptor antagonist irbesartan (10(-6) mol/L) pre-treatment; (4) Irbesartan plus AngII group: stimulation by AngII (10(-7) mol/L) with ROCK inhibitor Y27632 (10(-6) mol/L) pre-treatment. Then the products of protein and RNA were collected. Western blot and QuantiGene were used to detect the activation of RhoA-Rock pathway and CTGF. RESULTS: Exploring the affect of irbesartan on AngII through the Western blot analysis of CTGF and RhoA protein expression: the CTGF level was up-regulated by AngII (0.89 ± 0.05 vs control 0.48 ± 0.10, P < 0.01). Such an effect was markedly blocked by a pretreatment of irbesartan (0.72 ± 0.05, P < 0.05). After the use of AngII, the expression of RhoA protein was significantly enhanced (3.40 ± 0.46 vs control 1.77 ± 0.37, P < 0.01) and blunted by a pretreatment of irbesartan (2.27 ± 0.45, P < 0.05). The Western blot analysis of CTGF protein expression showed that AngII caused a robust increase in CTGF (0.62 ± 0.15 vs control 0.16 ± 0.05, P < 0.01). Such an effect was markedly blocked by a pretreatment of Y27632 (0.17 ± 0.04, P < 0.01). The result was similar at the gene level. AngII significantly increased the expression of CTGF mRNA (1.16 ± 0.06 vs control 1.00 ± 0.01, P < 0.01). And it was markedly blocked by a pretreatment of irbesartan (0.99 ± 0.07, P < 0.01) or Y27632 (1.04 ± 0.08, P < 0.05). AngII significantly increased the expression of RhoA mRNA (1.21 ± 0.07 vs control 1.00 ± 0.06, P < 0.01). And it was markedly blocked by a pretreatment of irbesartan (1.00 ± 0.12, P < 0.05) but not Y27632 (1.10 ± 0.05, P > 0.05). CONCLUSION: Ang II activates HFL-1 to produce CTGF through the AT-1 receptor. And the RhoA-Rock pathway is involved.

[Effect of toluene diisocyanate on reactive oxygen species production and permeability of human bronchial epithelial cells in vitro].

OBJECTIVE: To investigate the effect of toluene diisocyanate (TDI) on the production of reactive oxygen species (ROS) and the permeability of human bronchial epithelial (HBE) cells. METHODS: TDI-human serum albumin (TDI-HSA) conjugate was prepared using a modified Son's method. MTT assay was used to assess HBE cell viability after exposure to different concentrations of TDI-HSA. The level of intracellular ROS of HBE cells was detected by flow cytometry with an oxidation-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) uploading, and the permeability of cell monolayer was assessed by detecting the transepithelial electrical resistance (TEER). RESULTS: The exposure to 120 µg/ml TDI-HSA did not obviously affect the cell viability. Compared with the control group, the intracellular fluorescent intensity increased significantly in the cells exposed to 20, 60, and 100 µg/ml TDI-HSA (P<0.05). The intracellular ROS production increased significantly after 100 µg/ml TDI-HSA treatment (P<0.05), but the increment in ROS production was significantly suppressed by pretreatment of the cells with N-acetylcysteine (NAC) (P<0.05), which also enhanced the TEER decreased by TDI-HSA treatment (P<0.05). CONCLUSIONS: TDI enhances the permeability of HBE cell monolayer partially through a ROS-mediated pathway, suggesting the importance of oxidative stress in TDI-induced pulmonary diseases.

The generation of the endothelial specific cdc42-deficient mice and the effect of cdc42 deletion on the angiogenesis and embryonic development.

BACKGROUND: High microvascular permeability plays an essential role in pathological process of multiple diseases such as septic shock, acute lung injury and acute respiratory distress syndrome, and burns. Inhibiting hyperpermeability is significant for controlling these conditions. Cdc42, as a main member of the small Rho GTPase family, plays a critical role in controlling and regulating the endothelial junctional permeability. We aimed to generate and identify endothelial specific cdc42-deficient mice by the Cre/loxp recombination approach, for examination in an animal model of the contribution of the cdc42 gene in the microvascular barrier function. METHODS: We crossed cdc42(Flox/Flox) mice with mice expressing endothelial cell-specific Cre recombinase, and the offspring with the genotype cdc42(Flox/+)Tie2Cre(+/-) were back-crossed with the cdc42(Flox/Flox) mice. The cdc42(Flox/Flox)Tie2Cre(+/-) mice in the F2 generation were the target mice. If the cdc42 deficient mice did not survive, we would observe the cdc42 deficient mice embryos, and compare them with wild-type mice embryos. RESULTS: Cdc42(flox/+)Cre(+/-) mice were mated with the cdc42(Flox/Flox) mice and among the living offspring there were no cdc42(Flox/Flox)Cre(+/-) target mice. We found the endothelial special cdc42 deficient embryos at the E7.5-E16.5 stage. We observed that cdc42 deficient embryos were much smaller, had fewer vessels and were a little more swollen compared with the wild-type embryos. CONCLUSIONS: Endothelial specific knockout of cdc42 caused embryonic lethality and the mice did not survive to birth. The target embryos were much smaller, had fewer vessels and were a little more swollen compared with the wild-type embryos. These results demonstrated that the cdc42 plays an important role in development of embryos and in development of microvessels as well as microvascular permeability.

[Respiratory syncytial virus increases the expression and release of high mobility group Box-1 protein in the lung tissue of mice].

OBJECTIVE: To investigate the expression and release of high mobility group Box-1 protein (HMGB1) in the lung tissue of mice with respiratory syncytial virus (RSV) infection. METHODS: Eighteen mice were randomized into PBS control group, RSV group and RSV/ribavirin group. Seven days after RSV infection in the mice in the latter two groups, the bronchoalveolar lavage fluid (BALF) was collected for cell counting and classification, and the levels of IL-4, IFN-gamma and HMGB1 in the supernatants of the BALF were detected. The left lungs of the mice were harvested for pathological examination with HE staining, and the right lungs were taken for detecting the expression of HMGB1 by Western blotting. RESULTS: RSV induced a TH1 inflammation in the lung tissue as shown by significantly increased IFN-gamma and decreased IL-4 levels in the BALF. The total BALF cells, neutrophils and macrophages in the RSV group were significantly higher than those in the control group (P<0.05), and the cell counts were significantly decreased by ribavirin treatment (P<0.05). HE staining showed neutrophil and lymphocyte infiltration in the lumen and submucous layer of the airway in RSV group. The level of HMGB1 in the BALF significantly increased in the RSV group as compared with that in the control group (P<0.05), but was lowered by ribavirin treatment (P<0.05). The expression of the HMGB1 in the lung tissue significantly increased in the RSV group in comparison with that in the control group (P<0.05), and was not significantly decreased by ribavirin treatment (P>0.05). CONCLUSIONS: The increased expression and release of HMGB1 in the lung tissue of mice with RSV infection is probably involved in the development of RSV infection-related lung diseases.

[Hydrogen dioxide increases the expression of vascular endothelial growth factor in human bronchiolar epithelium cell via a PI3K-dependent pathway].

OBJECTIVE: To investigate the effect of hydrogen dioxide (H(2)O(2)) on the expression of vascular endothelial growth factor (VEGF) in human bronchiolar epithelial (HBE) cells. METHODS: MTT assay was used to assess HBE cell viability after exposure to different concentrations of H(2)O(2). VEGF/beta-actin gene fragments were amplified simultaneously by RT-PCR from the total HBE cell RNA, and VEGF protein expression in the cells was detected using ELISA. RESULTS: The exposure to 200 micromol/L H(2)O(2) did not obviously affected the cell viability. Compared with those in the control cell, VEGF165/beta-actin and VEGF189/beta-actin ratios were significantly increased in the cells after treatment with 50, 200, and 600 micromol/L H(2)O(2) (P<0.05). The protein expression of VEGF significantly increased after 50 micromol/L H(2)O(2) treatment (P<0.05), but significantly decreased with pretreatment with the PI3K inhibitor Ly294002 (P>0.05). CONCLUSION: Oxidative stress increases the expression of VEGF via a PI3K-dependent pathway in human bronchiolar epithelial cells, which may play an important role in the onset and maintenance of chronic inflammation in asthma.

[Lung cancer in pregnancy: report of two cases and review of literature].

OBJECTIVE: to raise awareness about lung cancer in pregnancy. METHODS: the clinical presentations, diagnosis and treatment of 2 cases of lung cancer in pregnancy were reported, and related literatures were reviewed. RESULTS: The first case was a 31-year-old pregnant woman at 34(th)-week gestation, who presented with right sided pleural effusion on a Chest X-ray film. A boy was delivered by Cesarean section at 35 weeks of gestation. Biopsy of the right-supraclavicular lymph node was performed simultaneously, and histopathological examination showed metastatic large cell lung cancer. Her respiratory condition worsened after the Caesarian section, and so mechanical ventilation, antibiotics and gefitinib were administered, but the treatment failed. She died on the 28(th) day after Caesarian section. The second case was a 28-year-old pregnant woman at the 27 week of gestation. PET showed right lung cancer with metastases to the pericardium, right pleura, liver and pelvic cavity. Bronchoscopic biopsy showed small-cell lung cancer. After pregnancy termination, the patient received 2 cycles of chemotherapy consisting of cisplatin and etoposide. The size of lesions decreased and the patient returned to the local hospital. CONCLUSIONS: lung cancer in pregnancy is a rare condition with poor prognosis. To improve the prognosis and prevent the metastasis to the fetus, systemic therapy should be considered, and meanwhile maternal advantage must be always weighed against possible embryo-fetal risks.

[Effect of ribavirin upon secretion of thymic stromal lymphopoietin in respiratory syncytial virus-induced asthma exacerbation of mice].

OBJECTIVE: To investigate the role of ribavirin (RIB) in treating respiratory syncytial virus (RSV)-induced asthma exacerbation of mice. METHODS: (1) Cell experiment: 32 flasks of human airway epithelial cell 16HBEs were randomly divided into four groups: RSV group, RSV/RIB group, RIB group and control group. 16HBEs were infected with RSV at a multiplicity of infection (MOI) of 2. RIB 50 microg/ml was added in culture medium and Western blot used to detect the production of thymic stromal lymphopoietin (TSLP) protein; (2) Animal experiment: 32 female BALB/c mice were randomly divided into four groups: Ovalbumin (OVA) group, OVA/RSV group, OVA/RSV/RIB group and control group. Mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into murine nasal cavity and RIB 10 mg/kg intramuscularly administered. BUXCO noninvasive murine lung function detection instrument was used to examine the airway response to metacholine; ELISA was used to detect IL-4, IL-5, IL-13 and IFN-gamma in murine serum and TSLP in supernatants of bronchoalveolar lavage fluid (BALF); murine lung specimens were stained with HE to observe inflammation and immunohistochemical technique was employed to observe the production of TSLP in murine airway epithelial cells. RESULTS: The cell experiment demonstrated the productions of TSLP protein in RSV group, RSV/RIB group, RIB group and control group were 1.97 +/- 0.22, 1.16 +/- 0.19, 0.99 +/- 0.17 and 0.89 +/- 0.08 respectively, and the production of TSLP in RSV/RIB group was lower than that in RSV group (P < 0.01). The animal experiment demonstrated that the murine airway responsiveness in RSV/OVA/RIB group was lower than that in OVA/RSV group (P < 0.01). The levels of IL-4 [(109.7 +/- 41.9) pg/ml], IL-5 [(220.8 +/- 30.9) pg/ml], IFN-gamma [(13.0 +/- 3.4) pg/ml] in murine serum and TSLP [(1945 +/- 82) pg/ml] in BALF of RSV/OVA/RIB group were significantly lower than those in OVA/RSV group [(274.2 +/- 103.7), (293.3 +/- 46.1), (30.1 +/- 5.7) and (2127 +/- 46) pg/ml respectively, all P < 0.01]; less infiltration of airway inflammatory cells in OVA/RSV/RIB group was observed than that in OVA/RSV group. Immunohistochemical staining of TSLP also showed a lower production of TSLP in airway epithelial cells of OVA/RSV/RIB group than OVA/RSV group. CONCLUSION: Ribavirin can inhibit the elevated production of TSLP after RSV infection and relieve RSV-induced asthma exacerbation in mice.

[Respiratory syncytial virus infection promotes the production of thymic stromal lymphopoietin and accelerates Th2 inflammation in mouse airway].

OBJECTIVE: To investigate the effect of respiratory syncytial virus (RSV) infection on the production of thymic stromal lymphopoietin (TSLP) and Th1/Th2 balance in asthmatic mice. METHODS: Thirty-two female BALB/c mice were randomly divided into 4 groups, namely the PBS group, ovalbumin (OVA) group, RSV group and OVA/RSV group. The mice were sensitized by OVA and then stimulated with nebulized OVA, and RSV was inoculated into the nasal cavity of the mice. BUXCO noninvasive lung function detection was performed to examine the airway response to metacholine, and enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of IL-4, IL-5, IL-13, and IFN-gamma in the mice. The cells in the bronchoalveolar lavage fluid (BALF) were counted and classified, and the supernatants of the BALF were used for the detection of TSLP. Histopathological changes in the lung tissues of the mice were examined using HE staining, and immunohistochemistry using anti-mouse TSLP antibody was performed to examine TSLP expressions in the airway epithelial cells. RESULTS: RSV infection promoted the production of TSLP in the asthmatic mice, and the concentration of TSLP in OVA/RSV group (2.13-/+0.05 ng/ml) was significantly higher than that in the other groups (P<0.01). RSV infection increased the serum levels of IL-4, IL-5, IL-13, and IFN-gamma in the mice. The total BALF cells, eosinophils, lymphocytes and neutrophils in OVA/RSV group were significantly higher than those in the other groups; noninvasive lung function examination showed higher Penh value in OVA/RSV group (318.66-/+50.87) than in the other groups when the inhaled metacholine increased to 6.25 mg/ml (P<0.01). More obvious and extensive airway inflammatory cell infiltration in OVA/RSV group were observed, and immunohistochemical staining also showed higher expression of TSLP in the airway epithelial cells of OVA/RSV group. CONCLUSIONS: RSV infection promotes the production of TSLP in the airway epithelial cells and increases the level of Th2 cytokines in asthmatic mice. Concurrent RSV infection can exacerbate Th2 inflammatory reaction in asthmatic mice.

[Causal analysis of initial misdiagnosis of pulmonary embolism].

OBJECTIVE: To analyze the causes of initial erroneous diagnosis of pulmonary embolism (PE) to improve the diagnostic efficiency. METHODS: The clinical data of 63 patients with a definite diagnosis of PE were retrospectively analyzed. According to the initial diagnosis, the patients were divided into definite diagnosis group (Group A, 23 cases) and misdiagnosis group (group B, 40 cases). The risk factors, initial symptoms, time of definite diagnosis, Wells scores, revised Geneva scores, and findings in chest X-ray and ECGs after onset and before the definite diagnosis were compared between the two groups. RESULTS: In group A, recent operations, malignancy, long-term bedridden state, PE history and deep vein thrombosis (DVT) symptom were more commonly seen than in group B, and the patients in group B were more likely to have hypertension, smoking, diabetes mellitus and lower limb varicose veins. The patients in group B had significantly lower Wells scores and revised Geneva scores than those in group A [2.50 (5.00) vs 6.00 (6.00), u=-3.296, P<0.001; 5.50 (4.75) vs 12.00 (9.00), u=-3.187, P<0.001, respectively]. In group B, chest examination in 22 of the 40 cases (55%) reported pulmonary infection, and among them, 15 were misdiagnosed as pneumonia. In groups A and B, SIQIIITIII/QIIITIII in ECG was found in 5 (21.7%) and 0 cases (0%), and normal ECG in 2 (8.7%) and 18 (45.0%) cases, respectively, showing significant difference between the two groups (P=0.010 and 0.003, respectively). CONCLUSION: The initial misdiagnosis of PE results mainly from the low awareness of some of the PE risk factors on the part of the physicians, atypical clinical manifestations and excessive dependence on chest films and ECGs.

[Risk factors of ventilator-associated pulmonary Candida infection and drug-resistant analysis].

OBJECTIVE: To investigate the risk factors for pulmonary Candida infection in association with mechanical ventilation and analyze the drug resistance profile of the Candida species that cause the infection. METHODS: A retrospective analysis was conducted 114 patients receiving mechanical ventilation for over 48 h. According to the presence of pulmonary Candida infections, these patients were divided into infected group (n=50, 43.9%) and non-infected group (64 cases). Univariate analysis and multivariate logistic regression analysis were performed to identify the risk factors for the infection, and drug sensitivity test was carried out to evaluate the drug resistance of the Candida species. RESULTS: Univariate analysis and multivariate logistic regression showed that the presence of at least two underlying diseases (OR=4.758, P=0.009), frequent changes of antibiotics (OR=6.128, P=0.001), and blood albumin below 25 g (OR=15.829, P=0.011) were the independent risk factors for pulmonary Candida infection associated with mechanical ventilation, and prophylactic antifungal treatment (OR=0.062, P=0.012) was a protective factor. Drug sensitivity test showed that Candida albicans was sensitive to most of the antifungal agents (100.0%), but the non-albicans Candida species were resistant to fluconazol (50.0%) and Itraconazole (38.5%). CONCLUSION: Poor general conditions and frequent changes of antibiotics are the major risk factors for pulmonary Candida infection in patients receiving mechanical ventilation. Drug resistant analysis is helpful in the treatment of the infections.

[Perindopril and losartan attenuate bleomycin A5-induced pulmonary fibrosis in rats].

OBJECTIVE: To evaluate the effects of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin II type 1 receptor (AT-1 receptor) blocker on the progression of rat pulmonary fibrosis induced by bleomycin A5. METHODS: Twenty-four male Wistar rats were randomized into pulmonary fibrosis model, perindopril treatment, losartan treatment and control groups. In the former 3 groups, pulmonary fibrosis was induced via intratracheal injection of bleomycin A5 (5 mg/kg), after which the rats in the perindopril and losartan groups received intragastric administration of the corresponding agents at the daily dose of 2 mg/kg and 10 m/kg, respectively. The rats in the control group had intratracheal injection of normal saline only. In the 4th week, the histological changes of the lung tissues were examined microscopically with Masson staining. Hydroxyproline content in the lungs was measured, and the protein expressions of AT-1 receptor, TGF-beta1 and IkappaBalpha were examined using Western blotting. DNA binding activity of NF-kappaB was analyzed with electrophoretic gel mobility shift assay (EMSA), and zymography was used to assess the activity of matrix metalloproteinase-2 and 9 (MMP-2, 9). RESULTS: Both perindopril and losartan treatment significantly reduced the pulmonary fibrosis score, content of hydroxyproline, protein expression of TGF-beta1, DNA binding activity of NF-kappaB and MMP-2, 9 activity, and increased cytoplasmic protein expression of IkappaBalpha. Perindopril treatment lowered the protein level of AT-1 receptor. CONCLUSION: Perindopril and losartan may inhibit bleomycin A5-induced pulmonary fibrosis in rats by reducing the protein expression of TGF-beta1 and suppressing the DNA binding activity of NF-kappaB and MMP-2, 9 activity.

[Effect of N-acetylcysteine on HMGB1 and RAGE expression in the lungs of asthmatic mice].

OBJECTIVE: To investigate the expression of HMGB1 and RAGE mRNA in the lungs of asthmatic mice and the effect of N-acetylcysteine (NAC) on their expression. METHODS: Twenty-one female BALB/c mice were randomly divided into control group, asthma group and NAC group (n=7). The expressions of HMGB1 and RAGE mRNA and their distributions in the lungs were detected by RT-PCR and immunohistochemical method. RESULTS: The expression levels of HMGB1 and RAGE mRNA were not significantly different between the control group (0.88-/+0.02 and 1.20-/+0.20, respectively) and the asthma model group (0.86-/+0.05 and 1.21-/+0.08, P>0.05). After NAC treatment, both of HMGB1 and RAGE mRNA levels (0.98-/+0.05 and 1.58-/+0.21) were significantly higher than those in the other two groups (P<0.05). HMGB1 was found in the nuclei and membrane of the bronchial and alveolar epithelial cells, and RAGE was located on the membrane of the alveolar epithelial cells. CONCLUSION: HMGB1 and RAGE may play a role in the oxidative stress during asthma, but the exact mechanism needs further investigation.