Comparative analysis of the lung microbiota in patients with respiratory infections, tuberculosis, and lung cancer: A preliminary study.

Recent evidence suggests that lung microbiota can be recognized as one of the ecological determinants of various respiratory diseases. However, alterations in the lung microbiota and associated lung immunity in these respiratory diseases remain unclear. To compare the lung microbiota and lung immune profiles in common respiratory diseases, a total of 78 patients were enrolled in the present study, including 21 patients with primary pulmonary tuberculosis (PTB), eight patients with newly diagnosed lung cancer (LC), and 49 patients with community-acquired pneumonia (CAP). Bronchoalveolar lavage fluid (BALF) was collected for microbiota and cytokine analyses. With MiSeq sequencing system, increased bacterial alpha-diversity and richness were observed in patients with LC than in those with PTB and CAP. Linear discriminant analysis effect size revealed that CAP-associated pulmonary microbiota were significantly different between the PTB and LC groups. More key functionally different genera were found in the PTB and LC groups than in the CAP group. The interaction network revealed stronger positive and negative correlations among these genera in the LC group than in the other two groups. However, increased BALF cytokine profiles were observed in the PTB group than in the other two groups, while BALF cytokines were correlated with key functional bacteria. This comparative study provides evidence for the associations among altered lung microbiota, BALF inflammation, and different respiratory disorders, which provides insight into the possible roles and mechanisms of pulmonary microbiota in the progression of respiratory disorders.

Radiofrequency ablation for lung squamous cell carcinoma in a single-lung patient: A case report and literature review.

RATIONALE: High morbidity and high mortality are the main features of non-small cell lung cancer (NSCLC). Radiofrequency ablation, which produces a large amount of heat to kill tumor cells, is one effective way to treat this disease. PATIENT CONCERNS: We report the case of a 74-year-old man who presented with a 1-month history of right chest pain. His left lung was removed 12 years prior. Chest computed tomography (CT) revealed a mass in the right lower lobe. DIAGNOSES: An excision biopsy of the mass showed lung squamous cell carcinoma. INTERVENTIONS: We performed radiofrequency ablation. OUTCOMES: The patient underwent 3.5 and 10 months of follow-up, with a partial response and complete remission, respectively. LESSONS: CT-guided radiofrequency ablation is a safe and an effective minimally invasive treatment option. Radiofrequency appears to be a valuable alternative to surgery for inoperable patients presenting with a single-lung NSCLC.

Tetrahydrocurcumin­induced autophagy via suppression of PI3K/Akt/mTOR in non­small cell lung carcinoma cells.

Lung carcinoma is the leading cause of mortality due to cancer worldwide. Autophagy has a significant role in the development and progression of non­small cell lung carcinoma (NSCLC). A previous study has revealed that tetrahydrocurcumin (THC), a traditional Chinese medicine isolated from Curcuma wenyujin (Chen & Ling, 1981), induces autophagy in human A549 NSCLC cells. The present study evaluated THC­induced autophagy in A549 cells using various assays, including the Cell Counting Kit­8, acridine orange staining, flow cytometry, reverse transcription­quantitative polymerase chain reaction (RT­qPCR), and western blot analysis of the markers of autophagy. THC inhibited the growth and proliferation of A549 cells (P<0.05). Acridine orange staining and flow cytometry revealed that THC treatment significantly enhanced autophagic cell proliferation inhibition (P<0.05). The RT­qPCR analysis revealed that THC treatment increased Beclin­1 expression level and compared with the control group (P<0.05). The light chain 3 (LC3)­II/LC3­I ratio was reduced in THC­treated cells when compared with the control group (P<0.05). Protein expression of various markers of autophagy, including p62, phosphorylated (p)­mechanistic target of rapamycin (mTOR), phosphoinositide 3­kinase (PI3K), p­PI3K, protein kinase B (Akt), and p­Akt was significantly reduced in THC­treated cells (P<0.05). In conclusion, the present study revealed the underlying mechanisms associated with THC­induced autophagy. A promising method of enhancing the therapeutic efficacy of THC against NSCLC cells may include inducing autophagy via inhibition of the PI3K/Akt/mTOR signaling pathway.

Small cell lung cancer treated by radiofrequency ablation: A case report.

RATIONALE: The morbidity and mortality of small cell lung cancer (SCLC), an uncommon malignancy of the lung, remain high. Radiofrequency ablation (RFA) creates heat to destroy cancer cells and is usually used to treat non-SCLC, but not SCLC. PATIENT CONCERNS: An 85-year-old male presented with a 2-month history of a productive cough with white phlegm and a 2-day history of hemoptysis. Chest computed tomography revealed a mass in the right lower lobe. DIAGNOSES: An excision biopsy of the mass showed SCLC. INTERVENTIONS: We treated the tumor with RFA. OUTCOMES: At the 2-year follow-up examination, the efficacy of the RFA was evaluated as a partial response. LESSONS: RFA can improve the prognosis of SCLC and should be considered for its treatment.

LARP1 is regulated by the XIST/miR-374a axis and functions as an oncogene in non-small cell lung carcinoma.

La-related protein 1 (LARP1) is a conserved RNA-binding protein and is known to regulate 5'-terminal oligopyrimidine tract (TOP) mRNA translation. Dysregulated LARP1 has been reported to be related to the development of several cancers. However, the exact function and mechanism of LARP1 in non-small cell lung cancer (NSCLC) is largely unknown. In the present study, we found that the mRNA levels of LARP1 were increased in NSCLC cells compared to those in normal control cells. Knockdown of LARP1 inhibited cell proliferation, migration and invasion in NSCLC cells and tumourigenicity in H520 cells. Both in vitro and in vivo analyses confirmed that STAT3 signalling was inactivated by the knockdown of LARP1. Moreover, LARP1 was identified as a direct target of miR-374a. Overexpression of miR-374a attenuated the promotor effects of LARP1 by inhibiting proliferation, metastasis and STAT3 signalling. Clinically, LARP1 was markedly overexpressed in NSCLC tissues, and upregulated LARP1 was correlated with tumour progression and poor survival. The expression of miR-374a was negatively correlated with the expression of LARP1 in NSCLC tissues. Furthermore, we found that XIST functioned as a competing endogenous RNA to suppress miR-374a, which regulated its downstream target LARP1. In summary, we suggest that the dysfunction of the XIST/miR-374a/LARP1 axis contributes to NSCLC and may serve as a promising therapeutic strategy for treatment.

Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis.

The early secreted protein early secretory antigenic target 6(ESAT-6) and the culture filtrate protein 10 (CFP-10) are 2 antigens that are specific to Mycobacterium tuberculosis. These 2 antigens are good targets for tuberculosis (TB) detection.To rapidly diagnose TB across a variety of samples, we developed colloidal gold immunochromatographic strips (ICSs) based on ESAT-6 and CFP-10.The strips were evaluated using 233 samples, including sputum, plasma, and pleural effusion samples.The positive detection rates for ICSs for ESAT-6 and CFP-10 in sputum (culture-positive for M tuberculosis) were 100% and 91.2%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in plasma were 34.1% and 29.4%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in pleural effusion were 64.7% and 55.9%, respectively. Experimental analysis of culture supernatant showing that the ICS developed for ESAT-6 had a sensitivity of 100% and a specificity of 91.2%. While the ICS developed for CFP-10 had a sensitivity of 91.2% and a specificity of 88.2%.The validity of the test is limited by source of sample. The technique is sensitive and specific for samples in sputum and culture media but not for plasma or pleural effusion samples. Detection of M tuberculosis using ICSs is rapid, simple, and relatively effective; thus, ICSs are a potential screening tool for TB.

Suppression of GRASP65 phosphorylation by tetrahydrocurcumin protects against cerebral ischemia/reperfusion injury via ERK signaling.

The aim of the present study was to assess the neuroprotective effects of tetrahydrocurcumin (THC) in a mouse model of cerebral ischemia/reperfusion (I/R) injury, and to investigate the involvement of Golgi reassembly and stacking protein 65 (GRASP65) and the extracellular signal­regulated kinase (ERK) signaling pathway. Cerebral I/R injury was induced using the Pulsinelli four­vessel occlusion method. After 5 min of reperfusion, mice received THC (5, 10 or 25 mg/kg) or saline by intraperitoneal injection. After 24 h of reperfusion, mice underwent neurological evaluation. Infarct volumes were determined by triphenyltetrazolium chloride staining, and levels of superoxide dismutase and malondialdehyde were measured in brain tissue homogenates. Expression of GRASP65, phosphorylated­GRASP65, ERK and phosphorylated­ERK was determined by western blotting. THC induced a dose­dependent decrease in the phosphorylation of ERK and GRASP65. Thus, THC attenuated I/R injury­induced activation of the ERK signaling pathway and reduced the phosphorylation of GRASP65. THC exhibited a dose­dependent protective effect against cerebral I/R injury, mediated by suppression of the ERK signaling pathway and a subsequent reduction in GRASP65 phosphorylation. The current study provided new information in the research of the cerebral ischemia­reperfusion injury mechanism.

Autophagy is involved in regulating the immune response of dendritic cells to influenza A (H1N1) pdm09 infection.

Autophagy can mediate antiviral immunity. However, it remains unknown whether autophagy regulates the immune response of dendritic cells (DCs) to influenza A (H1N1) pdm09 infection. In this study, we found that infection with the H1N1 virus induced DC autophagy in an endocytosis-dependent manner. Compared with autophagy-deficient Beclin-1(+/-) mice, we found that bone-marrow-derived DCs from wild-type mice (WT BMDCs) presented a more mature phenotype on H1N1 infection. Wild-type BMDCs secreted higher levels of interleukin-6 (IL-6), tumour necrosis factor- α (TNF-α), interferon-ß (IFN-ß), IL-12p70 and IFN-γ than did Beclin-1(+/-) BMDCs. In contrast to Beclin-1(+/-) BMDCs, H1N1-infected WT BMDCs exhibited increased activation of extracellular signal-regulated kinase, Jun N-terminal kinase, p38, and nuclear factor-κB as well as IFN regulatory factor 7 nuclear translocation. Blockade of autophagosomal and lysosomal fusion by bafilomycin A1 decreased the co-localization of H1N1 viruses, autophagosomes and lysosomes as well as the secretion of IL-6, TNF-α and IFN-ß in H1N1-infected BMDCs. In contrast to Beclin-1(+/-) BMDCs, H1N1-infected WT BMDCs were more efficient in inducing allogeneic CD4(+) T-cell proliferation and driving T helper type 1, 2 and 17 cell differentiation while inhibiting CD4(+) Foxp3(+) regulatory T-cell differentiation. Moreover, WT BMDCs were more efficient at cross-presenting the ovalbumin antigen to CD8(+) T cells. We consistently found that Beclin-1(+/-) BMDCs were inferior in their inhibition of H1N1 virus replication and their induction of H1N1-specific CD4(+) and CD8(+) T-cell responses, which produced lower levels of IL-6, TNF-α and IFN-ß in vivo. Our data indicate that autophagy is important in the regulation of the DC immune response to H1N1 infection, thereby extending our understanding of host immune responses to the virus.