Implication of m6A Methylation Regulators in the Immune Microenvironment of Bronchopulmonary Dysplasia.

N6-methyladenosine (m6A) regulates gene expression and governs many important biological processes. However, the function of m6A in the development of bronchopulmonary dysplasia (BPD) remains poorly characterized. Thus, the purpose of this investigation was to evaluate the effects of m6A RNA methylation regulators on the development of BPD. BPD-related transcriptome data were downloaded from the GEO database. Differentially expressed m6A methylation regulators between BPD and control group were identified. Consensus clustering was conducted for the classification of BPD and association between clusters and BPD phenotypes were explored. Analysis of differentially expressed genes (DEGs) and immune-related DEGs was performed. The GSEA, GO and KEGG analyses were used to interpret the functional enrichments. The composition of immune cell subtypes in BPD subsets was predicted by CIBERSORT analysis. Compared with the control group, expression of most m6A regulators showed significant alteration, especially for IGF2BP1/2/3. BPD was classified into 2 subsets, and cluster 1 was correlated with severe BPD. Furthermore, the results of functional enrichment analyses showed a disturbed immune-related signaling pathway. Based on CIBERSORT analysis, we found that the proportion of immune cell subsets changed between cluster 1 and cluster 2. Our study revealed the implication of m6A methylation regulators in the development of BPD, which might provide a novel insight for the diagnosis and treatment of BPD.

Establishment of a juvenile mouse asthma model induced by postnatal hyperoxia exposure combined with early OVA sensitization.

Objective: To establish a juvenile mouse asthma model by postnatal hyperoxia exposure combined with early ovalbumin (OVA) sensitization. Methods: Female C57BL/6J newborn mice were exposed to hyperoxia (95 % O2) from postnatal day-1 (PND1) to PND7; intraperitoneally injected with OVA suspension on PND21, PND28; and stimulated by nebulized inhalation of 1 % OVA from PND36 to PND42. Within 48 h of the last challenge, we observed their activity performance and evaluated airway responsiveness (AHR). All mice were executed at PND44. Female (n = 32) were divided into four groups as follows: room air（RA）+phosphate-buffered saline (PBS) group; O2 (hyperoxia, 95 % O2) + PBS group; RA + OVA group; O2+OVA group. We obtained the serum, bronchoalveolar lavage fluid (BALF), and lung tissues. The Wright-Giemsa staining was performed for leukocyte classification in BALF and HE staining for pathological examination. The levels of IL-2, IL-5, IL-13, IL-17A and IL-10 in BALF and tIgE and sIgE in serum were detected by ELISA. Results: Compared with OVA sensitization or hyperoxia exposure alone, the mice in the model group (O2+OVA) showed asthma-like symptoms and increased airway hyperreactivity,The levels of IL-5,IL-13 IL-17A were increased in BLAF,and total leukocyte and eosinophil counts were also significant increasesed. The levels of tIgE and sIgE in serum were increased. Conclusion: Postnatal hyperoxia exposure combined with early OVA sensitization might establish a juvenile mouse asthma model.

Recruitment of PVT1 Enhances YTHDC1-Mediated m6A Modification of IL-33 in Hyperoxia-Induced Lung Injury During Bronchopulmonary Dysplasia.

Bronchopulmonary dysplasia (BPD) is a chronic lung disease that specifically affects preterm infants. Oxygen therapy administered to treat BPD can lead to hyperoxia-induced lung injury, characterized by apoptosis of lung alveolar epithelial cells. Our epitranscriptomic microarray analysis of normal mice lungs and hyperoxia-stimulated mice lungs revealed elevated RNA expression levels of IL-33, as well as increased m6A RNA methylation levels of IL-33 and PVT1 in the hyperoxia-stimulated lungs. This study aimed to investigate the role of the PVT1/IL-33 axis in BPD. A mouse model of BPD was established through hyperoxia induction, and lung histological changes were assessed by hematoxylin-eosin staining. Parameters such as radial alveolar count and mean chord length were measured to assess lung function. Mouse and human lung alveolar epithelial cells (MLE12 and A549, respectively) were stimulated with hyperoxia to create an in vitro BPD model. Cell apoptosis was detected using Western blotting and flow cytometry analysis. Our results demonstrated that silencing PVT1 suppressed apoptosis in MLE12 and A549 cells and improved lung function in hyperoxia-stimulated lungs. Additionally, IL-33 reversed the effects of PVT1 both in vivo and in vitro. Through online bioinformatics analysis and RNA-binding protein immunoprecipitation assays, YTHDC1 was identified as a RNA-binding protein (RBP) for both PVT1 and IL-33. We found that PVT1 positively regulated IL-33 expression by recruiting YTHDC1 to mediate m6A modification of IL-33. In conclusion, silencing PVT1 demonstrated beneficial effects in alleviating BPD by facilitating YTHDC1-mediated m6A modification of IL-33. Inhibition of the PVT1/IL-33 axis to suppress apoptosis in lung alveolar epithelial cells may hold promise as a therapeutic approach for managing hyperoxia-induced lung injury in BPD.

Cryptotanshinone affects HFL-1 cells proliferation by inhibiting cytokines secretion in RAW264.7 cells and ameliorates inflammation and fibrosis in newborn rats with hyperoxia induced lung injury.

Objective: Bronchopulmonary dysplasia (BPD) is a common complication of prematurity and has no specific treatment option. Moreover, inflammation and fibrosis play a vital role in the development of BPD. Thus, this study aimed to explore the role of the anti-inflammatory and anti-fibrotic drug cryptotanshinone (CTS) in the treatment of inflammation and fibrosis in BPD. Methods: In vivo, Sprague-Dawley rats (male) were divided into air, hyperoxia and CTS groups with different dose interventions (7.5, 15, and 30 mg/kg). A BPD rat model was induced by continuous inhalation of hyperoxia (95%) for 7 days, during which different doses of CTS were injected intraperitoneally. Furthermore, histological examination, hydroxyproline content measurement, Western blot and real-time quantitative polymerase chain reaction were used to detect the levels of inflammation and fibrosis in the tissues. RAW264.7 cells exposed to 95% oxygen were collected and co-cultured with fibroblasts to determine the expression levels of α-SMA, collagen-Ⅰ and MMPs. The levels of pro-inflammatory cytokines such as TNF-α, IL-6 and pro-fibrotic factor TGF-ß1 in the supernatants were measured using enzyme-linked immunosorbent assay. Results: Haematoxylin and eosin staining revealed that CTS reduced the inflammatory response in rat lungs. Masson staining revealed that CTS alleviated the level of pulmonary fibrosis. CTS also reduced the levels of TNF-α, IL-6 and TGF-ß1 along with the expression of the fibrosis marker α-SMA in lung tissue. Similarly, in vitro analysis revealed that CTS decreased the levels of TNF-α, IL-6 and TGF-ß1 expressed in RAW 264.7 cells, and reduced α-SMA, collagen-Ⅰ, MMPs concentrations in HFL-1 cells co-cultured with the supernatant of RAW264.7 cells after hyperoxia. Conclusion: CTS can attenuate the hyperoxia-induced inflammatory response and the level of fibrosis by regulating the levels of inflammatory factors and fibrotic factor TGF-ß1 expressed by macrophages, thereby highlighting the therapeutic potential of CTS in the treatment of BPD.

Effects of early postnatal hyperoxia exposure combined with early ovalbumin sensitization on lung inflammation and bacterial flora in a juvenile mouse model of asthma.

Objective: The aim of this study is to explore the effects of early postnatal hyperoxia exposure combined with early ovalbumin (OVA) sensitization on lung inflammation and bacterial flora in neonatal mice on a juvenile mouse model of asthma. Methods: Thirty-two newborn female C57BL/6 J mice were randomly divided into four groups, which including room air+phosphate-buffered saline (PBS) group, hyperoxia+PBS group, room air+OVA group, and hyperoxia+OVA group, according to the hyperoxia exposure and/or OVA induction. Mice were exposed to either 95% O2 or room air for 7 days after birth; after 7 days, they were exposed to air and received an intraperitoneal injection of OVA suspension or PBS solution on postnatal days 21 (P21) and 28 (P28). From P36 to P42, the mice were allowed to inhale of 1% OVA or 0.9% NaCl solution. The mice were observed after the last excitation. HE staining was performed to observe the pathological changes in lung tissues. Wright-Giemsa staining was used to perform bronchoalveolar lavage fluid (BALF) leukocyte sorting. Enzyme-linked immunosorbent assay was used to determined the cytokines levels of interleukin (IL)-2, IL-5, IL-13, IL-17A, and IL-10 and serum IgE levels in BALF. Additionally, 16S rRNA sequencing was used to analyze the characteristics of lung microbiota. Results: Mice in the hyperoxia+OVA group showed asthma-like symptoms. HE staining results revealed a significant thickening of the airway wall and airway inflammation. BALF analysis of cellular components showed significant increases in total leukocyte and eosinophil counts and the levels of cytokines related to Th2 (IL-5 and IL-13) and Th17 (IL-17A); 16S rRNA sequencing revealed that the main members of the pulmonary microflora were Actinobacteriota, Proteobacteria, Firmicutes, and Bacteroidota at the phylum level. In addition, the bacteria with a major role were Acinetobacter and Moraxellaceae in the O2 + OVA group. Conclusion: The mouse suffering from postnatal hyperoxia exposure and early OVA sensitization, changes in symptoms, pathology, leukocyte and eosinophil counts, and levels of different T-cell cytokines in BALF and lung microbiota, which may provide a basis for the establishment of a juvenile mouse model of asthma.

Predictive and Diagnostic Values of Systemic Inflammatory Indices in Bronchopulmonary Dysplasia.

BACKGROUND: Bronchopulmonary dysplasia (BPD) is the most common respiratory complication in preterm infants, and there is a lag in the diagnosis of BPD. Inflammation is a vital pathogenic factor for BPD; we aim to evaluate the predictive and diagnostic values of systemic inflammatory indices in BPD. METHODS: Between 1 January 2019 and 31 May 2023, the clinical data of 122 premature infants with a gestational age of <32 weeks in the Department of Neonatology, the Affiliated Huai'an No. 1 People's Hospital of Nanjing Medical University, were retrospectively collected and classified into non-BPD (n = 72) and BPD (n = 50) groups based on the National Institute of Child Health and Human Development 2018 criteria. To compare the general characteristics of each group, we identified the independent risk variables for BPD using multivariate logistic regression analysis, compared the systemic inflammatory indices at birth, 72 h, 1 week, 2 weeks, and 36 weeks postmenstrual age (PMA), and constructed the receiver operating characteristic curves of neutrophil-to-lymphocyte ratio (NLR) diagnosis of BPD at different time points. RESULTS: ① The independent risk factors for BPD in preterm infants were birth weight, small for gestational age, and days of oxygen therapy (all p < 0.05). ② At 72 h and 1 week after birth, the serum NLR of the BPD group was higher than for the non-BPD group (p < 0.05). Furthermore, the neutrophil count (N), NLR, monocyte-to-lymphocyte ratio (MLR), systemic immune-inflammation index, systemic inflammation response index (SIRI), and pan-immune-inflammation value of infants with BPD were higher than the non-BPD group at 3 weeks after birth (p < 0.05). Moreover, at 36 weeks of PMA, the serum N, NLR, MLR, and SIRI of BPD infants were higher than those of non-BPD infants (p < 0.05). ③ The NLR of infants with and without BPD gradually increased after birth, reaching a peak at 72 h and 1 week, respectively. At 3 weeks postnatal, the NLR had the highest predictive power for BPD, with an area under the curve (AUC) of 0.717 (p < 0.001); the sensitivity was 56% and specificity was 86.1%. In addition, the NLR at 36 weeks of PMA exhibited some diagnostic value for BPD. The AUC was 0.693 (p < 0.001), the sensitivity was 54%, and specificity was 83.3%. CONCLUSIONS: At 3 weeks after birth and 36 weeks of PMA, some systemic inflammation indices (like N, NLR, SIRI) of preterm infants with BPD have specific predictive and diagnostic values; these indices may help the management of high-risk preterm infants with BPD.

Expression of long noncoding RNA uc.375 in bronchopulmonary dysplasia and its function in the proliferation and apoptosis of mouse alveolar epithelial cell line MLE 12.

Background: According to our previous gene ChIP results, long noncoding RNA uc.375 was down-regulated in lung tissue of bronchopulmonary dysplasia (BPD) mice induced by hyperoxia. FoxA1 gene showed higher levels in lung tissue of BPD mice and is reported to promote the apoptosis of alveolar epithelial cells. We aimed to clarify the expression pattern of uc.375 in BPD and explore the interaction between uc.375 and FoxA1. Methods: Newborn mice were placed in a 95% high-oxygen environment for 7 days. Lung tissue samples from mice were used for lncRNA microarray to screen BPD related lncRNAs. Mouse alveolar epithelial cell line MLE 12 was stably transfected with uc.375 and FoxA1 silencing or overexpression lentiviral vectors. The proliferation activity of MLE 12 cells was detected by a cell counting kit 8 (CCK-8) assay. MLE 12 cell apoptosis was determined by Hoechst/PI staining and flow cytometry analysis. The protein levels of Cleaved Caspase-3, FoxA1, SP-C and UCP2 were investigated by western blot. The relative mRNA expression levels were detected by quantitative real-time PCR. Results: uc.375 is mainly distributed in the nucleus of alveolar epithelial cells, as revealed by In Situ Hybridization assay results. uc.375 was lowly expressed in the lung tissues of BPD mice. According to the results of CCK-8 assay, analysis of Hoechst/PI staining and western blotting, uc.375 silencing inhibited cell proliferation, facilitated apoptosis of MLE 12 cells, promoted caspase 3 and FoxA1 expression, and inhibited the expression of SP-C and UCP2. On the contrary, after overexpressing uc.375, the opposite results were obtained. Silencing FoxA1 inhibited MLE 12 apoptosis, promoted proliferation, inhibited apoptosis-related factor caspase 3, and promoted the expression of SP-C and UCP2. FoxA1 silencing also reversed the effect induced by uc.375 knockdown on the proliferation and apoptosis of MLE 12 cells. Conclusion: Based on the biomedical images-derived analysis results, uc.375 negatively regulates FoxA1 expression, affects alveolar development, and plays an important role in the initiation and progression of BPD, providing a new molecular target for the prevention and treatment of BPD.

Role of the LRP1-pPyk2-MMP9 pathway in hyperoxia-induced lung injury in neonatal rats. / LRP1-pPyk2-MMP9é€šè·¯åœ¨é«˜æ°§è¯±å¯¼æ–°ç”Ÿå¤§é¼ è‚ºæŸä¼¤ä¸­çš„ä½œç”¨.

OBJECTIVES: To study the role of the low-density lipoprotein receptor-related protein 1 (LRP1)-proline-rich tyrosine kinase 2 phosphorylation (pPyk2)-matrix metalloproteinases 9 (MMP9) pathway in hyperoxia-induced lung injury in neonatal rats. METHODS: A total of 16 neonatal rats were randomly placed in chambers containing room air (air group) or 95% medical oxygen (hyperoxia group) immediately after birth, with 8 rats in each group. All of the rats were sacrificed on day 8 of life. Hematoxylin and eosin staining was used to observe the pathological changes of lung tissue. ELISA was used to measure the levels of soluble LRP1 (sLRP1) and MMP9 in serum and bronchoalveolar lavage fluid (BALF). Western blot was used to measure the protein expression levels of LRP1, MMP9, Pyk2, and pPyk2 in lung tissue. RT-PCR was used to measure the mRNA expression levels of LRP1 and MMP9 in lung tissue. RESULTS: The hyperoxia group had significantly higher levels of sLRP1 and MMP9 in serum and BALF than the air group (P<0.05). Compared with the air group, the hyperoxia group had significant increases in the protein expression levels of LRP1, MMP9, and pPyk2 in lung tissue (P<0.05). The hyperoxia group had significantly higher relative mRNA expression levels of LRP1 and MMP9 in lung tissue than the air group (P<0.05). CONCLUSIONS: The activation of the LRP1-pPyk2-MMP9 pathway is enhanced in hyperoxia-induced lung injury in neonatal rats, which may be involved in the pathogenesis of bronchopulmonary dysplasia.

Chest Computed Tomography Images in Neonatal Bronchial Pneumonia under the Adaptive Statistical Iterative Reconstruction Algorithm.

This study was to explore the application value of chest computed tomography (CT) images processed by artificial intelligence (AI) algorithms in the diagnosis of neonatal bronchial pneumonia (NBP). The AI adaptive statistical iterative reconstruction (ASiR) algorithm was adopted to reconstruct the chest CT image to compare and analyze the effect of the reconstruction of CT image under the ASiR algorithm under different preweight and postweight values based on the objective measurement and subjective evaluation. 85 neonates with pneumonia treated in hospital from September 1, 2015, to July 1, 2020, were selected as the research objects to analyze their CT imaging characteristics. Subsequently, the peripheral blood of healthy neonates during the same period was collected, and the levels of C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were detected. The efficiency of CT examination, CRP, ESR, and combined examination in the diagnosis of NBP was analyzed. The results showed that the subjective quality score, lung window subjective score, and mediastinal window subjective score were the highest after CT image reconstruction when the preweight value of the ASiR algorithm was 50%. After treatment, 79 NBP cases (92.9%) showed ground-glass features in CT images. Compared with the healthy neonates, the levels of CRP and ESR in the peripheral blood of neonates with bronchial pneumonia were much lower (P < 0.05). The accuracy rates of CT examination, CRP examination, ESR examination, CRP + ESR examination, and CRP + ESR + CT examination for the diagnosis of NBP were 80.7%, 75.3%, 75.1%, 80.3%, and 98.6%, respectively. CT technology based on AI algorithm showed high clinical application value in the feature analysis of NBP.

A clinical study on plasma biomarkers for deciding the use of adjuvant corticosteroid therapy in bronchopulmonary dysplasia of premature infants.

Objective: The study was designed to investigate some plasma markers which help us to decide the use of adjuvant corticosteroid therapy in bronchopulmonary dysplasia (BPD) of premature infants. Methods: Thirty BPD infants were treated by dexamethasone. Among these cases, dexamethasone was significant effective in 10 cases, and no significant effective in 20 cases. These patients were divided into two groups as the significant effect (SE) group (n=10) and the non-significant effect (NE) group (n=20) according to the curative effect of dexamethasone. Fifteen non-BPD infants with gestational age and gender matching were selected as the control group. Plasma samples before and after dexamethasone treatment were collected from three infants chosen randomly from SEG for the data-independent acquisition (DIA) analysis. ELISA was further used to detect the levels of differential proteins LRP1 and S100A8 in all individuals, including SE, NE and control groups. Results: DIA analysis results showed that after dexamethasone treatment, there were a total of 52 plasma proteins that showed significant differences, of which 43 proteins were down-regulated and 9 proteins were up-regulated. LRP1 and S100A8 were two plasma proteins that were significantly changed after dexamethasone treatment. Compared with the control group, plasma LRP1 was significantly increased in BPD. Interestingly, the plasma concentration of LRP1 in the NE group was significantly higher than that in the SE group. S100A8, as an indicator of plasma inflammation, was significantly higher in BPD than the control group. Unlike LRP1, there was no significantly difference between the SE and NE group (P=0.279) before dexamethasone treatment. Conclusion: Elevated plasma LRP1 and S100A8 in BPD infants are two indicators that correlated with the efficacy of dexamethasone, and might be used as biomarkers for deciding the use of adjuvant corticosteroids therapy in the BPD.

Proteomic analysis of sex differences in hyperoxic lung injury in neonatal mice.

Sex-specific differences in the severity of bronchopulmonary dysplasia (BPD) are due to different susceptibility to hyperoxic lung injury, but the mechanism is unclear. In this study, neonatal male and female mouse pups (C57BL/6J) were exposed to hyperoxia and lung tissues were excised on postnatal day 7 for histological analysis and tandem mass tags proteomic analysis. We found that the lung sections from the male mice following postnatal hyperoxia exposure had increased alveolar simplification, significant aberrant pulmonary vascularization and arrest in angiogenesis compared with females. Comparison of differentially expressed proteins revealed 377 proteins unique to female and 425 unique to male as well as 750 proteins in both male and female. Bioinformatics analysis suggested that several differentially expressed proteins could contribute to the differences in sex-specific susceptibility to hyperoxic lung injury. Our results may help identify sex-specific biomarkers and therapeutic targets of BPD.

High versus Low Mechanical Index Imaging Diagnostic Ultrasound in Patients with Myocardial Infarction: A Therapeutic Application Study.

BACKGROUND High mechanical index impulse of ultrasound is used for diagnosis of microvascular coronary obstruction and the necrotic area, but an experimental model study suggested that it can restore microvascular and epicardial coronary flow. The purposes of the study were to test the safety and therapeutic efficacy of high acoustic energy diagnostic ultrasound in patients with ST-segment elevation myocardial infarction. MATERIAL AND METHODS Patients with ST-segment elevation myocardial infarction subjected to a low (n=199) or high (n=251) mechanical index ultrasound before and after percutaneous coronary interventions and echocardiographic parameters were evaluated. Coronary angiographies were performed for the assessment of culprit vessels. Thrombolysis in myocardial infarction flow grade 1 or 2 were considered as culprit vessels. RESULTS Patients diagnosed through low acoustic energy ultrasound reported 235 infarct vessels and patients diagnosed through high acoustic energy ultrasound reported 300 infarct vessels. With respect to low acoustic energy, high acoustic energy reduced the number of culprit vessels at post-percutaneous coronary interventions at 48 hours before hospital discharge (P=0.015) and post-percutaneous coronary interventions at 1-month from the baseline interventions (P=0.043). Also, the maximum% ST-segment resolution and an ejection fraction of the left ventricle was increased and microvascular coronary obstruction in infarct vessels was decreased for both evaluation points. High acoustic energy could not affect heart rate (P=0.133) and oxygen saturation (P=0.079). CONCLUSIONS High acoustic energy ultrasound is a safe method for diagnosis of ST-segment elevation myocardial infarction and may have therapeutic applications.

Optical coherence tomography versus intravascular ultrasound in patients with myocardial infarction: a diagnostic performance study of pre-percutaneous coronary interventions.

Accurate coronary measurements are important in guiding percutaneous coronary intervention. Intravascular ultrasound is a widely accepted diagnostic modality for coronary measurement before percutaneous coronary intervention. The spatial resolution of optical coherence tomography is 10 times larger than that of intravascular ultrasound. The objective of the study was to compare quantitative and qualitative parameters of frequency domain optical coherence tomography (FDOCT) with those of intravascular ultrasound and coronary angiography in patients with acute myocardial infarction. Diagnostic parameters of coronary angiography, intravascular ultrasound, and FDOCT of 250 patients with coronary artery disease who required admission diagnosis were included in the analyses. Minimum lumen diameter detected by FDOCT was larger than that detected by quantitative coronary angiography (2.11±0.1 vs 1.89±0.09 mm, P<0.0001, q=34.67) but smaller than that detected by intravascular ultrasound (2.11±0.1 vs 2.19±0.11 mm, P<0.0001, q=12.61). Minimum lumen area detected by FDOCT was smaller than that detected by intravascular ultrasound (3.41±0.01 vs 3.69±0.01 mm2, P<0.0001). FDOCT detected higher numbers of thrombus, tissue protrusion, dissection, and incomplete stent apposition than those detected by intravascular ultrasound (P<0.0001 for all). More accurate and sensitive results of the coronary lumen can be detected by FDOCT than coronary angiography and intravascular ultrasound (level of evidence: III).

Optical coherence tomography versus intravascular ultrasound in patients with myocardial infarction: a diagnostic performance study of pre-percutaneous coronary interventions

Accurate coronary measurements are important in guiding percutaneous coronary intervention. Intravascular ultrasound is a widely accepted diagnostic modality for coronary measurement before percutaneous coronary intervention. The spatial resolution of optical coherence tomography is 10 times larger than that of intravascular ultrasound. The objective of the study was to compare quantitative and qualitative parameters of frequency domain optical coherence tomography (FDOCT) with those of intravascular ultrasound and coronary angiography in patients with acute myocardial infarction. Diagnostic parameters of coronary angiography, intravascular ultrasound, and FDOCT of 250 patients with coronary artery disease who required admission diagnosis were included in the analyses. Minimum lumen diameter detected by FDOCT was larger than that detected by quantitative coronary angiography (2.11±0.1 vs 1.89±0.09 mm, P<0.0001, q=34.67) but smaller than that detected by intravascular ultrasound (2.11±0.1 vs 2.19±0.11 mm, P<0.0001, q=12.61). Minimum lumen area detected by FDOCT was smaller than that detected by intravascular ultrasound (3.41±0.01 vs 3.69±0.01 mm2, P<0.0001). FDOCT detected higher numbers of thrombus, tissue protrusion, dissection, and incomplete stent apposition than those detected by intravascular ultrasound (P<0.0001 for all). More accurate and sensitive results of the coronary lumen can be detected by FDOCT than coronary angiography and intravascular ultrasound (level of evidence: III).

Effects of pilose antler peptide on bleomycin-induced pulmonary fibrosis in mice.

The aim of present study was to evaluate the effects of the pilose antler peptide (PAP) on bleomycin (BLM)-induced lung fibrosis. The lung wet-to-dry weight (W/D) ratio and myeloperoxidase (MPO) activity were measured. The serum levels of super-oxide dismutase (SOD), malondialdehyde (MDA) were examined. Then the contents of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 were determined using ELISA method. Furthermore, the protein expressions of Rho, ROCK1, p-IκB, IκB, p-NF-κB, NF-κB in lung tissues were detected by western blot analysis. As a result, PAP markedly decreased pulmonary W/D ratio, lung MPO activity and relieved lung histopathological changes. In addition, PAP increased the level of SOD and reduced the levels of MDA, TNF-α, IL-1ß, IL-6 in serum of BLM-stimulated mice. In addition, PAP remarkably inhibited the protein levels of ROCK/NF-κB pathway. In conclusion, our results showed that PAP exhibited protective effects on pulmonary fibrosis via the regulation of ROCK/NF-κB pathway.

All-trans retinoic acid upregulates the expression of ciliary neurotrophic factor in retinal pigment epithelial cells.

Retinopathy of prematurity, a leading cause of visual impairment in low birth-weight infants, remains a crucial therapeutic challenge. Ciliary neurotrophic factor (CNTF) is a promyelinating trophic factor that promotes rod and cone photoreceptor survival and cone outer segment regeneration in the degenerating retina. Ciliary neurotrophic factor expression is regulated by many factors such as all-trans retinoic acid (ATRA). In this study, we found that ATRA increased CNTF expression in mouse retinal pigment epithelial (RPE) cells in a dose- and time-dependent manner, and PKA signaling pathway is necessary for ATRA-induced CNTF upregulation. Furthermore, we showed that ATRA promoted CNTF expression through CREB binding to its promoter region. In addition, CNTF levels were decreased in serum of retinopathy of prematurity children and in retinal tissue of oxygen-induced retinopathy mice. In mouse RPE cells cultured with high oxygen, CNTF expression and secretion were decreased, but could be recovered after treatment with ATRA. In conclusion, our data suggest that ATRA administration upregulates CNTF expression in RPE cells.

[Expression of long non-coding RNA NANCI in lung tissues of neonatal mice with hyperoxia-induced lung injury and its regulatory effect on NKX2.1].

OBJECTIVE: To investigate the expression of long non-coding RNA NANCI in lung tissues of neonatal mice with hyperoxia-induced lung injury and its regulatory effect on NKX2.1. METHODS: A total of 48 neonatal C57BL/6J mice were randomly divided into an air group and a hyperoxia group, with 24 mice in each group. Each group was further divided into 7-day, 14-day, and 21-day subgroups, with 8 mice in each subgroup. The mice in the air group were fed in the indoor environment (FiO2=21%) and those in the hyperoxia group were fed in a high-oxygen box (oxygen concentration: >95%). The mice were sacrificed at each time point and lung tissue samples were collected. Hematoxylin and eosin staining was used to observe pathological changes in lung tissues. RT-qPCR and Western blot were used to measure the mRNA and protein expression of NANCI and NKX2.1. RESULTS: The air group had the highest mRNA expression of NANCI and NKX2.1 at 7 days and the same level of mRNA expression at 14 and 21 days. Compared with the air group, the hyperoxia group had significant reductions in the degree of alveolarization and radial alveolar count (RAC) in lung tissues (P<0.05), and in the hyperoxia group, RAC gradually decreased over the time of hyperoxia exposure (P<0.05). The hyperoxia group had significantly lower mRNA and protein expression of NANCI and NKX2.1 than the air group at all time points (P<0.05). In both groups, the relative mRNA and protein expression of NANCI and NKX2.1 gradually decreased over the time of hyperoxia exposure (P<0.05). The expression of NKX2 was positively correlated with that of NANCI (r=0.585, P=0.003), and the expression of NKX2 and NANCI was positively correlated with RAC in the hyperoxia group (r=0.655 and 0.541 respectively, P<0.05). CONCLUSIONS: NANCI may be involved in the development of immature lung tissues. Lung injury is gradually aggravated over the time of hyperoxia exposure. The levels of NANCI and NKX2.1 are associated with the severity of lung injury, suggesting that the NANCI/NKX2.1 target gene signaling pathway might be involved in the development of hyperoxia-induced lung injury in neonatal mice.

Fraction of exhaled nitric oxide and soluble receptors for advanced glycation end products are negatively correlated in children with recurrent wheezing.

BACKGROUND: The fraction of exhaled nitric oxide (FeNO) and serum levels of the soluble receptor for advanced glycation end products (sRAGE) have been suggested as biomarkers for asthma. OBJECTIVE: This study aimed to assess the correlation between FeNO and sRAGE serum levels in children <5 years old with recurrent wheezing. METHOD: In total, 88 children with recurrent wheezing were divided into the high-risk group or low-risk group according to their clinical features. The high-risk group included 60 children, 42 male and 18 female, average age 36.7 months (range 32-48.7 months); the low-risk group included 28 children, 20 male and 8 female, average age 38.1 months (range 33-46.2 months).Asthma in high-risk children was treated with aerosol inhalation of Pulmicort respules 1 mg/d for four continuous weeks, while asthma in low-risk children was treated with symptomatic treatment. FeNO, serum sRAGE and eosinophils (EOS) were examined by ELISA and a regular blood cell analyzer. RESULTS: The serum sRAGE level was 738±191 and 992.4±210 pg/ml and the mean FeNO level was 27.3 and 17.6 ppm, respectively, in the asthma high-risk and low-risk group, showing significant differences between the two groups. In addition, FeNO and sRAGE serum levels were negatively correlated.After the inhalation of Pulmicort respules, FeNO decreased and sRAGE increased, while EOS showed no significant change. CONCLUSIONS: FeNO and sRAGE serum levels are negatively correlated in children with recurrent wheezing. Further larger scale studies are needed to test the use of FeNO and sRAGE as biomarkers for the prediction of asthma in children.

Differential expression of long non-coding RNAs in hyperoxia-induced bronchopulmonary dysplasia.

Bronchopulmonary dysplasia (BPD) is a common complication of premature birth that seriously affects the survival rate and quality of life among preterm neonates. Long non-coding RNAs (lncRNAs) have been implicated in many human diseases. However, the role of lncRNAs in the pathogenesis of BPD remains poorly understood. Here, we exposed neonatal C57BL/6J mice to 95% concentrations of ambient oxygen and established a mouse lung injury model that mimicked human BPD. Next, we compared lncRNA and messenger RNA (mRNA) expression profiles between BPD and normal lung tissues using a high-throughput mouse lncRNA + mRNA array system. Compared with the control group, 882 lncRNAs were upregulated, and 887 lncRNAs were downregulated in BPD lung tissues. We validated some candidate BPD-associated lncRNAs by real-time quantitative reverse-transcription polymerase chain reaction analysis in eight pairs of BPD and normal lung tissues. Gene ontology, pathway and bioinformatics analyses revealed that a downregulated lncRNA, namely AK033210, associated with tenascin C may be involved in the pathogenesis of BPD. To the best of our knowledge, our study is the first to reveal differential lncRNA expression in BPD, which provides a foundation for further understanding of the molecular mechanism of BPD development. Copyright © 2016 John Wiley & Sons, Ltd.