miR­186­5p regulates the inflammatory response of chronic obstructive pulmonary disorder by targeting HIF­1α.

Chronic obstructive pulmonary disorder (COPD) is a chronic respiratory disease that is a major cause of morbidity and mortality worldwide. Previous studies have shown that miR­186­5p expression is significantly increased in COPD and is involved in multiple physiological and pathological processes. However, the role of miRNA­186­5p in the inflammatory response of COPD remains unclear. In this study, an in vitro model of COPD was established using lipopolysaccharide (LPS)­induced human bronchial epithelial cells (BEAS­2B). CCK­8 assays, flow cytometry, and a Muse cell analyzer were used to determine cell viability, cell cycle distribution, and apoptosis, respectively. The production of TNF­α and IL­6 were measured by ELISA. Reverse­transcription­quantitative PCR and western blotting were used to analyze mRNA and protein expression levels. The targeting relation between miR­186­5p and HIF­1α was discovered using dual­luciferase reporter assays. The results showed that transfection of miR­186­5p inhibitor inhibited cell proliferation and promoted cell apoptosis in the LPS­induced BEAS­2B cells. Inhibition of miR­186­5p markedly increased the levels of TNF­α and IL­6. miR­186­5p directly targeted and negatively regulated HIF­1α expression. In addition, inhibition of miR­186­5p increased the expression of the NF­κB pathway protein p­p65. In conclusion, it was found that inhibiting miR­186­5p may improve inflammation of COPD through HIF­1α in LPS­induced BEAS­2B cells, possibly by regulating NF­κB signaling. These findings provide a novel potential avenue for the clinical management of COPD. Future research is required to determine the mechanism of the interaction between miR­186­5p and HIF­1α in COPD.

LncRNA GAS5 regulated by FTO-mediated m6A demethylation promotes autophagic cell death in NSCLC by targeting UPF1/BRD4 axis.

Long non-coding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) has been shown to be a regulator for many cancers, including non-small cell lung cancer (NSCLC). Therefore, its role and mechanism in the process of NSCLC deserve to be further revealed. The expression levels of GAS5, fat mass and obesity-associated protein (FTO) and bromodomain-containing protein 4 (BRD4) were detected by quantitative real-time PCR. Western blot analysis was used to examine the protein expression of FTO, BRD4, up-frameshift protein 1 (UPF1) and autophagy-related markers. Methylated RNA immunoprecipitation was used to assess the m6A level of GAS5 regulated by FTO. Cell proliferation and apoptosis were determined using MTT assay, EdU assay and flow cytometry. Autophagy ability was assessed by immunofluorescence staining and transmission electron microscope. Xenograft tumor model was constructed to explore the effects of FTO and GAS5 on NSCLC tumor growth in vivo. The interaction between UPF1 and GAS5 or BRD4 was confirmed by pull-down assay, RIP assay, dual-luciferase reporter assay, and chromatin immunoprecipitation. Fluorescent in situ hybridization was used to analyze the co-localization of GAS5 and UPF1. Actinomycin D treatment was employed to evaluate BRD4 mRNA stability. GAS5 was downregulated in NSCLC tissues and was associated with poor prognosis in NSCLC patients. FTO was highly expressed in NSCLC, and it inhibited GAS5 expression by reducing GAS5 m6A methylation level. GAS5 suppressed by FTO could promote the autophagic death of NSCLC cells in vitro and inhibit NSCLC tumor growth in vivo. In addition, GAS5 was able to interact with UPF1 to reduce the mRNA stability of BRD4. Knockdown of BRD4 reversed the inhibition of GAS5 or UPF1 silencing on the autophagic cell death of NSCLC. The findings of the study showed that lncRNA GAS5 mediated by FTO could contribute to the autophagic cell death of NSCLC by interacting with UPF1 to reduce BRD4 mRNA stability, suggesting that GAS5 might be a vital therapy target for NSCLC progression.

MiRNA-423 rs6505162 and miRNA-6811 rs2292879 SNP associated with lung cancer in Hainan, China.

BACKGROUND: MicroRNAs (miRNAs) are known to exert significant influence on various physiological processes and diseases, including cancers. The primary objective of this present study was to examine the impact of eight single-nucleotide polymorphisms (SNPs) in miRNA on the susceptibility to lung cancer (LC) within the Chinese Southern population. METHODS: The genotypes of these eight polymorphisms were determined in 132 LC patients and 214 cancer-free controls. RESULTS: In overall analyses, GG genotype of miRNA-6811 rs2292879 polymorphism was significantly correlated with increased risk of LC (GG vs. AA, adjusted OR = 5.10, 95% CI = 1.02-25.43, P=0.047), yet the genotype frequencies of rs2292879 SNP in controls did not met the Hardy-Weinberg equilibrium (HWE) (P=0.001) in present study. Stratified analyses by smoking revealed that miRNA-423 rs6505162 variants significantly decreased the LC risk in heterozygous (CA vs. CC, adjusted OR = 0.14, 95% CI = 0.03-0.81, P=0.028) and recessive (AA vs. CA + CC, adjusted OR = 0.17, 95% CI = 0.03-0.90, P=0.038) genetic models in smoking population. However, miRNA-196A2 rs11614913, miRNA-196A2 rs12304647, miRNA-146A rs2910164, miRNA-16-1 rs1022960, miRNA-608 rs4919510, and miRNA-27a rs895819 polymorphisms were not significantly associated with LC. CONCLUSION: The findings of our study indicate a potential decrease in LC risk among smokers with the miRNA-423 rs6505162 variants, while an increase in risk is associated with miRNA-6811 rs2292879 polymorphisms in the population of Southern Chinese. However, further well-designed research is necessary to fully understand the precise impact of these two SNPs on the development of LC.

Simulation of Microscopic Seepage Characteristics of Interphase Mass Transfer in CO2 Miscible Flooding under Multiphysics Field Coupling Conditions.

CO2 miscible flooding in low permeability reservoirs is conducive to significantly improving oil recovery. At present, the microscopic displacement simulation of CO2 miscible flooding is mainly reflected in the simulation of the seepage process, but the pressure control of the seepage process is lacking, and the simulation of the characterization of CO2 concentration diffusion is less studied. In view of the above problems, a numerical model of CO2 miscible flooding is established, and the microscopic seepage characteristics of interphase mass transfer in CO2 miscible flooding are analyzed by multiphysics field coupling simulations at the two-dimensional pore scale. The injection velocity, contact angle, diffusion coefficient, and initial injection concentration are selected to analyze their effects on the microscopic seepage characteristics of CO2 miscible flooding and the concentration distribution in the process of CO2 diffusion. The research shows that after injection into the model, CO2 preferentially diffuses into the large pore space and forms a miscible area with crude oil through interphase mass transfer, and the miscible area expands continuously and is pushed to the outlet by the high CO2 concentration area. The increase in injection velocity will accelerate the seepage process of CO2 miscible displacement, which will increase the sweep area at the same time. The increase in contact angle increases the seepage resistance of CO2 and weakens the interphase mass transfer with crude oil, resulting in a gradual decrease in the final recovery efficiency. When the diffusion coefficient increases, the CO2 concentration in the small pores and the parts that are difficult to reach at the model edge will gradually increase. The larger the initial injection concentration is, the larger the CO2 concentration in the large pore and miscible areas in the sweep region at the same time. This study has guiding significance for the field to further understand the microscopic seepage characteristics of CO2 miscible flooding under the effect of interphase mass transfer.

Diagnostic value and safety of medical thoracoscopy under local anesthesia for unexplained diffuse interstitial lung disease: A retrospective study.

OBJECTIVE: We aimed to explore the safety and diagnostic value of medical thoracoscopic lung biopsy in patients with unexplained diffuse interstitial lung disease (ILD) in a single center pilot study. METHOD: We retrospectively analyzed clinical and pathological diagnostic data from 52 patients with diffuse ILD undergoing medical thoracoscopic lung biopsy. RESULTS: Forty-four cases of diffuse ILD were confirmed pathologically, giving a diagnostic rate of 84.6%. Among these 44 patients, 11 patients were diagnosed with cancer, including eight patients with lung adenocarcinoma, three patients with metastases; two from a gastrointestinal malignancy, and one from a granulosa cell tumor of the ovary. There were 17 cases of idiopathic interstitial pneumonia, including nine cases of usual interstitial pneumonia (UIP), four cases of non-specific interstitial pneumonia (NSIP), three cases of cryptogenic organizing pneumonia (COP), and one case of acute interstitial pneumonia (AIP). There were 12 cases of rare interstitial pneumonias, which included six cases of pulmonary alveolar proteinosis, one case each of pulmonary Langerhans cell histiocytosis (LCH) and pulmonary lymphangiomyomatosis, two cases of nodular sarcoidosis, and two cases of chronic eosinophilic pneumonia. We recorded various complications, including bleeding, infection, and pneumothorax. A total of 28 patients (53.8%) experienced at least one of the above complications, but there were no deaths associated with biopsy. CONCLUSIONS: Medical thoracoscopic lung biopsy appears a safe and effective method for diagnosing diffuse ILD of unknown cause but further prospective studies, with larger numbers, including comparison with other established techniques are required.

Role of Genetic Polymorphisms in IL12Rß2 in Chronic Obstructive Pulmonary Disease.

Background: Chronic obstructive pulmonary disease (COPD) is the most common chronic inflammatory airway disease. Il-12r beta 2 (IL-12Rß2) is important for the production of pathogenic Th1 cells. We aimed to explore the association between IL-12Rß2 genetic variants and COPD risk among southern Chinese Han population. Methods: We recruited 996 participants to perform an association analysis through SNPStats online software. We used false-positive report probability analysis to detect whether the positive findings were noteworthy. Haploview 4.2 software and SNPStats were used to conduct the haplotype analysis and linkage disequilibrium. Finally, the interaction of SNP-SNP in COPD risk was evaluated by multi-factor dimensionality reduction. Results: The study found evidence that genetic loci in IL-12Rß2 (rs2201584, rs1874791, rs6679356, and rs3790567) were potentially associated with the COPD susceptibility. In particular, IL-12Rß2-rs2201584 and -rs1874791 showed close associations with COPD risk in both overall and several stratified analyses. Overall analysis or several stratified analyses indicated that allele A or homozygous genotype AA of IL-12Rß2-rs2201584 were risk factors for COPD (Allele A: OR (95% CI) = 1.23 (1.02-1.48), p = 0.033; genotype AA: OR (95% CI) = 1.76 (1.15-2.69), p = 0.009). The allele A or homozygous genotype AA of IL-12Rß2- rs1874791 were also risk factors for COPD (Allele A: OR (95% CI) = 1.36 (1.10-1.68), p = 0.004; genotype AA: OR (95% CI) = 2.17 (1.18-3.99), p = 0.013). Conclusion: Intronic variants in IL-12Rß2 (rs2201584, rs1874791, rs6679356, and rs3790567) were associated with the COPD susceptibility. In particular, there were sufficient evidences that IL-12Rß2-rs2201584 and -rs1874791 were associated with the increasing risk of COPD.

PGC-1α attenuates TNF-α-induced inflammatory responses in OCCM-30 cells.

BACKGROUND AND OBJECTIVES: Peroxisome proliferator-activated receptor-γ coactivator (PGC)-1α, a master regulator of mitochondrial biogenesis and oxidative metabolism, has been associated with many inflammatory diseases. However, little is known about the function and mechanism of PGC-1α in cementoblasts under periodontitis. Our study aimed to investigate the effects of PGC-1α in immortalized cementoblast cell line OCCM-30 under TNF-α stimulation. MATERIALS AND METHODS: OCCM-30 cells were cultured and exposed to TNF-α, and PGC-1α expression was assessed by Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Chemical inhibitors targeting various signaling pathways including NF-κB, p38 MAPK, Akt, and p53 were used to identify the regulatory mechanism involved. ZLN005 was used to upregulate PGC-1α and the subsequent alteration of inflammatory cytokines expression under TNF-α stimulation were examined by qRT-PCR and Elisa. PGC-1α siRNA was employed to further verify the role of PGC-1α in inflammatory response. Dual-reporter gene assays were performed to examine the transcriptional activity of p65, and the phosphorylation level of p65 was evaluated by western blotting. Immunofluorescence assays and nuclear and cytoplasmic extractions were performed to check the nuclear translocation of p65. Coimmunoprecipitation studies were also performed to check whether there is direct binding between p65 and PGC-1α. RESULTS: TNF-α suppressed PGC-1α expression in OCCM-30 cells. Blocking p38 MAPK pathways restored the expression of PGC-1α. ZLN005 can upregulate PGC-1α in OCCM-30 cells. The upregulation of PGC-1α by ZLN005 inhibited TNF-α-induced proinflammatory cytokine expression, which was impaired by the transfection of PGC-1α siRNA. Knocking down PGC-1α also partially restored the ZLN005-decreased transcriptional activity of p65. However, the phosphorylation level and nuclear translocation of p65 were not significantly affected by PGC-1α. It was found that p65 was bound to PGC-1α in OCCM-30 cells stimulated by TNF-α, and the binding was increased upon ZLN005 treatment. CONCLUSIONS: PGC-1α can attenuate TNF-α-induced inflammatory responses in OCCM-30 cells.

IL6R gene polymorphisms and their relation to chronic obstructive pulmonary disease susceptibility in the Chinese population.

Background: This work was designed to explore the correlation between IL6R polymorphisms and chronic obstructive pulmonary disease (COPD) susceptibility. Methods: Agena MassARRAY was used to genotype five SNPs of IL6R in 498 patients with COPD and 498 controls. Genetic models and haplotype analysis were used to assess the associations between SNPs and COPD risk. Results: Rs6689306 and rs4845625 increase the risk of COPD. Rs4537545, rs4129267 and rs2228145 were related to a decreased risk of COPD in different subgroups. Haplotype analysis revealed that GTCTC, GCCCA and GCTCA contributed to a reduced risk of COPD after adjustment. Conclusion: IL6R polymorphisms are significantly associated with COPD susceptibility.

YAP1 expression in nasal polyps and its relationship with epithelial mesenchymal transition.

OBJECTIVE: This study explored and analyzed the expression of YAP1 in nasal polyps and its correlation with the epithelial-mesenchymal transition (EMT). METHOD: 58 patients with chronic sinusitis and nasal polyps, who were hospitalized in the otorhinolaryngology department of our hospital from January 2019 to May 2020 were recruited as the study cohort and placed in a nasal-polyp group, and, at the same time, another 30 nasal septum deviation with inferior turbinate hypertrophy patients were placed in a control group. The expressions of the YAP1 gene in the nasal polyp and turbinate mucosa tissues (using the immunohistochemical method), the YAP1 mRNA, E-cadherin mRNA, and vimentin mRNA expressions (using the RT-PCR method), and the YAP1, E-cadherin, and vimentin protein expressions (using Western blot) were measured, and the correlations between YAP1 and the expressions of E-cadherin and vimentin were analyzed. RESULTS: The immunohistochemistry results revealed that the positive rate of YAP1 expression in the nasal-polyps group was critically higher than the YAP1 expression in the control group (P<0.05). According to the RT-PCR results, the YAP1 mRNA and vimentin mRNA relative expression levels in the nasal-polyps group were significantly higher than they were in the control group (P<0.05), but the E-cadherin mRNA relative expression level in the nasal-polyp group was notably lower than it was in the control group (P<0.05). Our Western blot analysis showed that the protein expressions of YAP1 and vimentin protein in the nasal-polyps group were significantly higher than the corresponding protein expressions in the control group (P<0.05), but the E-cadherin expression in the nasal-polyp group was especially lower than it was in the control group (P<0.05). In addition, in the nasal polyp tissues, the relative expression between the YAP1 mRNA and the E-cadherin mRNA reflected a notably negative correlation (P<0.05), but the YAP1 mRNA and vimentin mRNA showed a positive correlation (P<0.05). CONCLUSION: High expressions of YAP1 and EMT occur in nasal polyp tissues, and YAP1 is likely to be involved in the regulation of EMT and might be one of the mechanisms in nasal polyps.

LncRNA GAS5 expression in non-small cell lung cancer tissues and its correlation with Ki67 and EGFR.

OBJECTIVE: This research explored and analyzed LncRNA GAS5 expression in non-small cell lung cancer (NSCLC) tissues and its correlation with Ki67 and EGFR. METHODS: A total of 130 samples of paraffin-embedded NSCLC tissues and para-cancerous normal tissues that were collected in the Department of Pathology from January 2014 to April 2016 were selected. The relative expression of LncRNA GAS5 and Ki67/EGFR in both NSCLC tissues and para-cancerous normal tissues were detected via RT-PCR and immunohistochemistry respectively. Subsequently, we analyzed the relative expression of LncRNA GAS5, the expression of Ki67/EGFR and its correlation with clinicopathological features and prognosis of patients, and studied the correlation between LncRNA GAS5 and Ki67/EGFR. RESULTS: The relative expression of LncRNA GAS5 in NSCLC tissues was substantially less than that of the para-cancerous normal tissues (P<0.05). The positive expression rate of Ki67/EGFR in NSCLC tissues remarkably exceeded that in para-cancerous normal tissues (P<0.05). The relative expression of LncRNA GAS5 was correlated with the degree of tumor differentiation, TNM staging and lymph node metastasis (P<0.05). The positive expression rate of Ki67 and EGFR in NSCLC tissues was related to TNM stage and metastasis of lymph node (P<0.05). In addition, the survival of patients with high LncRNA GAS5 expression was obviously superior to those with low LncRNA GAS5 expression (P<0.05), patients with negative Ki67 had superior survival than those with positive Ki67 (P<0.05), and patients with negative EGFR had increased survival over those with positive EGFR (P<0.05). Moreover, the positive rates of Ki67 and EGFR in patients with low LncRNA GAS5 expressions were obviously higher than those with high LncRNA GAS5 expressions (P<0.05). The relative expression level of LncRNA GAS5 in NSCLC patients had a remarkably negative correlation with Ki67 and EGFR (P<0.05). CONCLUSION: The decrease in LncRNA GAS5 expression and the over-express of Ki67/EGFR occur in NSCLC tissues, the expressions of LncRNA GAS5, Ki67 and EGFR are connected with the progression, metastasis and prognosis of tumor; and LncRNA GAS5 is related to the expression of Ki67 and EGFR. These three factors are involved in the tumorigenesis and growth of the NSCLC process.

Assessment and review of treatment for secondary spontaneous pneumothorax using medical thoracoscopy-assisted argon plasma coagulation in association with autologous blood pleurodesis.

AIMS: To evaluate the efficacy of medical thoracoscopy-assisted argon plasma coagulation in association with autologous blood pleurodesis in spontaneous pneumothorax. PATIENTS AND METHODS: Three male patients with spontaneous pneumothorax were treated; medical thoracoscopy-assisted argon plasma coagulation combined with autologous blood pleurodesis was conducted for all patients whose duration of the air leak exceeded 7 days. We systematically reviewed all of the relevant literature to analyze and sum up the treatments of secondary spontaneous pneumothorax. RESULTS: The air leaks were all sealed and no recurrence of pneumothorax was reported. No complications of fever, bleeding, or signs of infection were observed during the process. CONCLUSION: The authors believe that the combination of medical thoracoscopy-assisted argon plasma coagulation and autologous blood pleurodesis is safe and effective. However, due to the number of patients included in this uncontrolled case study, more cases will be collected in the future.The reviews of this paper are available via the supplemental material section.

Hypermethylation of the transmembrane protein125 in lung adenocarcinoma mediates the activation of NF- κB signaling pathway and reduces the sensitivity to decitabine / 中国肿瘤生物治疗杂志

@#［摘要］ 目的：探讨穿膜蛋白125（transmembrane protein125，TMEM125）在肺腺癌组织与A549 细胞中的表达，以及影响细胞 的增殖和侵袭能力的分子机制。方法：从癌症基因组图谱（the cancer genome atlas，TCGA）数据库收集肺腺癌数据包，下载临床 信息及基因表达谱数据。分析TMEM125 在肺腺癌组织中的表达及其与患者总生存期的相关性。构建TMEM125 过表达A549 细胞株，以CCK-8 法、细胞划痕实验检测TMEM125 过表达对肿瘤细胞的增殖和迁移能力的影响；流式细胞术检测TMEM125 过 表达对A549 细胞的细胞周期和凋亡的影响。WB检测TMEM125 过表达对下游NF-κB信号通路、凋亡蛋白的影响；免疫共沉淀 法（co-immunoprecipitation，Co-IP）检测TMEM125 与NF- κB 抑制因子结合Ras 样2（NF- κB inhibitor interacting Ras-like 2， NKIRAS2）的相互作用。利用TNFα（10 ng/ml）处理TMEM125 过表达A549 细胞，CKK-8、流式细胞术及WB检测其对细胞增殖、 凋亡以及NF-κB信号通路相关蛋白表达的影响。去甲基化试剂地西他滨处理A549 细胞，qPCR和WB检测TMEM125 基因和蛋 白的表达。结果：TMEM125 mRNA在肺腺癌组织中表达水平显著低于正常组织（P<0.001），启动子甲基化水平显著高于正常组 织（P<0.001），并且低、中表达患者总生存期显著低于高表达患者（P<0.001）。过表达TMEM125 抑制了A549 细胞的增殖和迁移 （P<0.01），增加细胞G2/M 期，促进细胞凋亡（P<0.01）；过表达TMEM125 与NKIRAS2 相互作用，显著抑制NF- κB 的活性 （P<0.01）；地西他滨处理A549 细胞可促进TMEM125 表达并且抑制细胞增殖（P<0.01）。结论：启动子高甲基化水平降低了 TMEM125 基因表达，导致其抑制NF-κB活性功能和抑制细胞增殖的作用下降，并且降低了细胞对地西他滨的敏感性。

Etiology and management of nasopharyngeal hemorrhage after radiotherapy for nasopharyngeal carcinoma.

OBJECTIVE: To analyze the etiology of nasopharyngeal hemorrhage after radiotherapy for nasopharyngeal carcinoma (NPC) and evaluate the relevant management and rescue approaches. METHODS: Seventeen cases of nasopharyngeal hemorrhage caused by radiotherapy of NPC, treated between January 2015 and March 2018, were retrospectively analyzed to study the etiology of nasopharyngeal hemorrhage. The management and rescue strategies, including anterior and posterior nostril packing, endoscopic nasopharynx electrocoagulation, and digital subtraction angiography embolization, were assessed for their effectiveness. RESULTS: Nasopharynx hemorrhage after radiotherapy of NPC was mainly associated with erosion of the internal carotid artery or maxillary artery by the tumor. Among the 17 cases, 11 patients were treated by digital subtraction arterial angiography embolization, and 3 were treated by endoscopic nasopharynx electrocoagulation. Overall, 13 patients survived, while 4 died. CONCLUSION: Anterior and posterior nostril packing, endoscopic nasopharynx electrocoagulation, and digital subtraction angiography embolization are suitable for treating nasopharyngeal hemorrhage. However, effective hemostasis depends on early identification of the bleeding vessels.

Association between TLR4 A299G polymorphism and pneumonia risk: a meta-analysis.

BACKGROUND: Several genetic studies have evaluated the association between Toll-like receptor 4 (TLR4) A299G polymorphism and the risk of pneumonia. However, the results were not consistent. We thus did this meta-analysis. MATERIAL AND METHODS: Relevant studies were systematically searched by using the NCBI, Medline, Web of Science, and Embase databases. Data were extracted independently by 2 investigators. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were estimated. RESULTS: Eight case-control studies with 658 patients and 1862 controls were included in this meta-analysis. TLR4 A299G polymorphism was significantly associated with pneumonia risk (OR=1.74; 95% CI 1.19-2.53; P=0.004). The result was significant in adults. In addition, TLR4 A299G polymorphism was also associated with community-acquired pneumonia (CAP) risk. Results from cumulative meta-analysis and sensitivity analysis suggested that the results are reliable and robust. CONCLUSIONS: The results of this meta-analysis suggest that susceptibility to pneumonia was associated with TLR4 A299G polymorphism.