Deubiquitylase USP31 Induces Autophagy and Promotes the Progression in Lung Squamous Cell Carcinoma Cells by Stabilizing E2F1 Expression.

BACKGROUND: Autophagy exerts a vital role in the progression of lung squamous cell carcinoma (LUSC). Ubiquitin-specific peptidase 31 (USP31) has recently been found to be involved in the development of a variety of cancers. However, whether USP31 modulates autophagy in LUSC remains unclear. METHODS: This study revealed that high levels of USP31 were discovered in LUSC tissue samples employing the Gene Expression Profiling Interactive Analysis (GEPIA) database, quantitative real- time PCR (qRT-PCR), and Western blot analysis. Cell proliferation was tested via cell counting kit 8 (CCK-8) as well as colony formation, demonstrating that USP31-stable knockdown reduced cell viability. RESULTS: Immunofluorescence analysis illustrated that USP31 knockdown blocked the occurrence of LUSC autophagy. Meanwhile, USP31 has been shown to stabilize the expression of E2F transcription factor 1 (E2F1) through the proteasome pathway. Furthermore, overexpressed E2F1 effectively eliminated the effect of USP31 knockdown on LUSC cell proliferation and autophagy. CONCLUSION: In summary, this investigation proved that USP31 promoted LUSC cell growth and autophagy, at least in part by stabilizing E2F1 expression, which provided a potential therapeutic gene for the treatment of LUSC.

Effect and mechanism of iron-carbon micro-electrolysis pretreatment of organic peroxide production wastewater.

The wastewater from organic peroxide production has high chemical oxygen demand (COD) concentration and poor biodegradability, so it is necessary to find a cost-effective treatment method. The iron-carbon microelectrolysis (IC-ME) technology was used to pretreat the organic peroxide production wastewater, and the influence of reaction conditions on the removal effect of pollutants and the degradation mechanism were studied. The effects of initial pH, iron filings, iron-carbon ratio, and reaction time on the wastewater treatment were investigated by single-factor and response surface optimization experiments, and the degradation mechanism was analyzed by three-dimensional fluorescence spectroscopy, UV-Vis, and gas chromatography mass spectrometry (GC-MS). The experimental results showed that the COD removal efficiency was 35.67% and the biodegradability of wastewater was increased from 0.113 to 0.173 under the conditions of initial pH of 3.1, the dosage of iron filings of 30.5 g/L, the ratio of iron-carbon of 1.01, and the reaction time of 122.8 min, and the process of IC-ME for degrading COD of wastewater from the production of organic peroxide was consistent with the secondary reaction. The IC-ME process could decompose macromolecular organic compounds such as tyrosine proteins and aromatic proteins, and improve the biodegradability of wastewater. It provides a theoretical reference for the practical application of IC-ME to treat this type of wastewater.

NLRP3 inflammasome mediates abnormal epithelial regeneration and distal lung remodeling in silica­induced lung fibrosis.

NOD-like receptor protein 3 (NLRP3) inflammasome is closely related to silica particle­induced chronic lung inflammation but its role in epithelial remodeling, repair and regeneration in the distal lung during development of silicosis remains to be elucidated. The present study aimed to determine the effects of the NLRP3 inflammasome on epithelial remodeling and cellular regeneration and potential mechanisms in the distal lung of silica­treated mice at three time points. Pulmonary function assessment, inflammatory cell counting, enzyme­linked immunosorbent assay, histological and immunological analyses, hydroxyproline assay and western blotting were used in the study. Single intratracheal instillation of a silica suspension caused sustained NLRP3 inflammasome activation in the distal lung. Moreover, a time­dependent increase in airway resistance and a decrease in lung compliance accompanied progression of pulmonary fibrosis. In the terminal bronchiole, lung remodeling including pyroptosis (membrane­distributed GSDMD+), excessive proliferation (Ki67+), mucus overproduction (mucin 5 subtype AC and B) and epithelial­mesenchymal transition (decreased E­Cadherin+ and increased Vimentin+), was observed by immunofluorescence analysis. Notably, aberrant spatiotemporal expression of the embryonic lung stem/progenitor cell markers SOX2 and SOX9 and ectopic distribution of bronchioalveolar stem cells were observed in the distal lung only on the 7th day after silica instillation (the early inflammatory phase of silicosis). Western blotting revealed that the Sonic hedgehog/Glioma­associated oncogene (Shh/Gli) and Wnt/ß­catenin pathways were involved in NLRP3 inflammasome activation­mediated epithelial remodeling and dysregulated regeneration during the inflammatory and fibrotic phases. Overall, sustained NLRP3 inflammasome activation led to epithelial remodeling in the distal lung of mice. Moreover, understanding the spatiotemporal profile of dysregulated epithelial repair and regeneration may provide a novel therapeutic strategy for inhalable particle­related chronic inflammatory and fibrotic lung disease.

Inhibition of Bruton's Tyrosine Kinase Alleviates Monocrotaline-Induced Pulmonary Arterial Hypertension by Modulating Macrophage Polarization.

Macrophage accumulation and activation contribute to the development of pulmonary arterial hypertension (PAH), while Bruton's tyrosine kinase (BTK) is an important regulator for the activation and polarization of macrophage. However, the role of BTK in PAH remains unknown. In the present study, a selective BTK inhibitor (BTKi) BGB-3111 was applied to investigate the role of BTK in monocrotaline- (MCT-) induced PAH rat and phorbol myristate acetate- (PMA-) differentiated U937 macrophages. Our results showed that BTK was mainly distributed and upregulated in CD68+ macrophages in the lungs of PAH rats. Daily treated with BTKi BGB-3111 alleviated MCT-induced PAH, as indicated by the decrease in right ventricular systolic pressure (RVSP), attenuation in right ventricle hypertrophy and pulmonary vascular remodeling, reduction in perivascular collagen deposition, as well as inhibition of inflammation and endothelial-to-mesenchymal transition (EndMT) in the lung. Moreover, BTK inhibition suppressed MCT-induced recruitment of macrophages, especially the classical activated macrophages (M1) in the lung. In vitro, BGB-3111 significantly suppressed lipopolysaccharide- (LPS-) induced M1 polarization and proinflammatory cytokine production in U937-derived macrophages. The underlying mechanism is associated with the inhibition of NF-κB/MAPK pathways and nucleotide-binding oligomerization domain-like receptor with pyrin domain 3 (NLRP3) inflammasome activation. Furthermore, macrophage conditioned medium (CM) from LPS-induced M1 macrophages promoted migration and EndMT of HPAECs, while CM from BGB-3111-pretreated LPS-induced M1 macrophages failed to induce this response. These findings suggest that BTK inhibition alleviates PAH by regulating macrophage recruitment and polarization and may be a potential therapeutic strategy for the treatment of PAH.

Paeoniflorin attenuates monocrotaline-induced pulmonary arterial hypertension in rats by suppressing TAK1-MAPK/NF-κB pathways.

Pulmonary arterial hypertension (PAH) characterized by pulmonary vascular remodeling is a lethal disease. Paeoniflorin (PF) is a monoterpene glycoside with numerous beneficial functions, such as vasodilation, anti-inflammation and immunomodulation. This study aims to investigate the effects of PF on monocrotaline (MCT)-induced PAH rats. Our data showed that both prophylactic or therapeutic administration of PF alleviated MCT-induced increasing of right ventricular systolic pressure (RVSP), prevented right ventricle hypertrophy and pulmonary arterial remodeling, as well as inhibited inflammatory cell infiltration around pulmonary arteries. Meanwhile, PF blocked MCT-induced endothelial-mesenchymal transition (EndMT) as indicated by the restored expression of endothelial markers in lung. Moreover, PF inhibited MCT-induced down-regulation of bone morphogenetic protein receptor 2 (BMPR2) and suppressed MCT-induced phosphorylation of transforming growth factor-ß (TGFß) activated kinase 1 (TAK1) in vivo. In vitro studies indicated that PF prevented human pulmonary arterial smooth muscle cells (PASMCs) from platelet-derived growth factor-BB (PDGF-BB)-stimulated proliferation and migration. PF also partially reversed TGFß1, interleukin-1ß (IL-1ß) and tumor necrosis factor (TNF-α) co-stimulated endothelial-to-mesenchymal transition (EndMT) in cultured human pulmonary artery endothelial cells (HPAECs). Signaling pathway analysis demonstrated that the underlying mechanism might be associated with the inhibition of TAK1-MAPK/NF-κB pathways. Taken together, our results suggested that PF could be a potential drug for the treatment of PAH.

The Effects of Lipoaspirate-Derived Fibrous Tissue on Survival Quality and Mechanical Property of Fat Grafts.

BACKGROUND: Fibrous connective tissue (referred to as fiber) in lipoaspirates would be discarded before lipotransfer in case of cannula blockage. However, the fiber contains extracellular matrix which provide structure support and is rich in stromal vascular fractions (SVFs). Removal of the fiber might theoretically affect the survival quality and mechanical properties of fat grafts. But there is few evidence in vivo and vitro about how the fiber affects the fat grafts. OBJECTIVE: To assess the effect of fibers on the survival quality and mechanical property of fat grafts. METHODS: The SVFs in both fat and fiber were obtained by collagenase digestion for cells counting and comparison. Three groups were designed according to the different proportions of fat and fiber: the fat group (100% fat), the mixed group (50% fat, 50% fiber in volume ratio), and the fiber group (100% fiber). Three groups of grafts were transplanted in vivo to evaluate the differences in volume retention rate, histological characteristics and mechanical properties. RESULTS: The amount of SVF cells in fibers (3.47 ±â€Š1.49 × 104 cells/mL) was significantly lower than that in fat (12.3 ±â€Š4.95 × 104 cells/mL) (P < 0.05). Grafts in the mixed group and the fiber group showed an increase of volume retention at week 4, but the fiber content showed no significant effects on the volume retention of grafts in three groups at week 12. Elasticity modulus of grafts in the fat group was higher than that in the fiber group and the mixed group at week 4 and 8, the fiber content showed no significant effects on the elasticity modulus of grafts in three groups at week 12. The addition of the fiber reduced the inflammation, cysts, fibrosis, and capillaries density of the grafts. CONCLUSIONS: There were few SVF cells in the fiber. When it was mixed with fat in different proportions and transplanted in vivo, the content of fiber showed no significantly different effects on the long-term volume retention and mechanical property of fat grafts. Due to the risk of blockage, it is recommended to discard the fiber in lipoaspirates.

The Efficacy and Safety of Subcutaneous Radiofrequency After Liposuction: A New Application for Face and Neck Skin Tightening.

BACKGROUND: Minimally invasive or noninvasive skin-tightening procedures have become trends in facial and neck rejuvenation. Radiofrequency-assisted liposuction (RFAL) is a new choice for the treatment of skin relaxation that is more effective than noninvasive surgery without surgical incision. OBJECTIVES: The authors recommend a 2-step method in which radiofrequency is applied after appropriate liposuction is performed. This approach is safer and more effective than traditional RFAL, and the authors detail the safety guidelines, operative techniques, postoperative satisfaction results, and complications. METHODS: A total of 227 patients with lower face and neck skin laxity underwent RFAL between April 2012 and June 2019. The following data were collected: age, body mass index, operative duration, volume of fat aspirated, amount of energy delivered, and number and type of complications. Patient satisfaction was surveyed postoperatively and assessed by third-party surgeons at 3 and 6 months. RESULTS: At 6 months after operation, 78.8% of patients considered the results moderate to excellent, whereas 21.2% of the patients considered the results to be poor or thought there was no change. The photograph evaluation performed by independent plastic surgeons showed moderate to excellent results in 89.1% of patients. There were no major complications that required further medical or surgical intervention. CONCLUSIONS: This 2-step method is a safe and effective improvement in the application of radiofrequency for face and neck skin tightening. Patients can achieve significant contour correction via minimally invasive surgery with a lower risk of side effects.

Characterization of MET exon 14 alteration and association with clinical outcomes of crizotinib in Chinese lung cancers.

BACKGROUND: Most studies on MET exon 14 (MET-ex14) alteration, defined as an oncogenic driver, have been carried out among Caucasians; similar studies among Chinese people are limited. METHODS: We retrospectively analyzed the genomic profiles of 11,306 Chinese patients with various stages of lung cancer to investigate the prevalence of MET-ex14. Survival outcomes were analyzed in evaluable patients who received front-line crizotinib (n = 44) or chemotherapy (n = 14). RESULTS: MET-ex14 alterations were identified in 125 patients, a frequency of 1.1 %, which is much lower than that in Caucasians (∼2.7 %). We found that MET-ex14 alterations were more likely to be detected in older patients (median age 69.0 years, p <0.001). Among evaluable patients harboring MET-ex14 alterations, longer progression-free survival (PFS) was observed with crizotinib than with chemotherapy (8.5 months versus 4.0 months, p = 0.041), but there was no difference in overall survival (OS, 11.3 months versus 12.0 months, p = 0.66). No significant difference in PFS or OS was found among MET splice-site variants or when there were concurrent TP53 alterations. Concurrent MET amplification results in a shorter PFS (4.2 months versus 8.5 months, p = 0.029) but a comparable OS (7.8 months versus 14.0 months, p = 0.12). Patients with undetectable baseline plasma MET-ex14 had a trend of longer PFS (p = 0.097) but comparable OS (p = 0.18). A novel MET Y1003C mutation was detected and demonstrated a clinical response to crizotinib. CONCLUSIONS: Our study demonstrated a prevalence of 1.1 % for MET-ex14 alterations among the Chinese population. Our study also contributes to a better understanding of molecular factors that are associated with clinical outcomes of patients with MET exon 14 alterations.

Multiplexed molecular profiling of lung cancer with malignant pleural effusion using next generation sequencing in Chinese patients.

Lung cancer is the most common type of cancer and the leading cause of cancer-associated death worldwide. Malignant pleural effusion (MPE), which is observed in ~50% of advanced non-small cell lung cancer (NSCLC) cases, and most frequently in lung adenocarcinoma, is a common complication of stage III-IV NSCLC, and it can be used to predict a poor prognosis. In the present study, multiple oncogene mutations were detected, including 17 genes closely associated with initiation of advanced lung cancer, in 108 MPE samples using next generation sequencing (NGS). The NGS data of the present study had broader coverage, deeper sequencing depth and higher capture efficiency compared with NGS findings of previous studies on MPE. In the present study, using NGS, it was demonstrated that 93 patients (86%) harbored EGFR mutations and 62 patients possessed mutations in EGFR exons 18-21, which are targets of available treatment agents. EGFR L858R and exon 19 indel mutations were the most frequently observed alterations, with frequencies of 31 and 25%, respectively. In 1 patient, an EGFR amplification was identified and 6 patients possessed a T790M mutation. ALK + EML4 gene fusions were identified in 6 patients, a ROS1 + CD74 gene fusion was detected in 1 patient and 10 patients possessed a BIM (also known as BCL2L11) 2,903-bp intron deletion. In 4 patients, significant KRAS mutations (G12D, G12S, G13C and A146T) were observed, which are associated with resistance to afatinib, icotinib, erlotinib and gefitinib. There were 83 patients with ERBB2 mutations, but only two of these mutations were targets of available treatments. The results of the present study indicate that MPE is a reliable specimen for NGS based detection of somatic mutations.

Icotinib Attenuates Monocrotaline-Induced Pulmonary Hypertension by Preventing Pulmonary Arterial Smooth Muscle Cell Dysfunction.

BACKGROUND: Aberrant activation of epidermal growth factor receptor (EGFR) signaling pathway is associated with the pathogenesis of pulmonary hypertension (PH). However, the effect of icotinib, a first generation of EGFR tyrosine kinase inhibitor (EGFR-TKI), on PH remains to be elucidated. METHODS: PH rat model was established by a single intraperitoneal injection of monocrotaline (MCT, 60 mg/kg). Icotinib (15, 30, and 60 mg/kg/day) was administered by oral gavage from the day of MCT injection. After 4 weeks, hemodynamic parameters and histological changes of the pulmonary arterial vessels were assessed, and the phenotypic switching of pulmonary arterial smooth muscle cells (PASMCs) was determined in vivo. Moreover, the effects of icotinib (10 µM) on epidermal growth factor (EGF, 50 ng/ml)-stimulated proliferation, migration, and phenotypic switching of human PASMCs were explored in vitro. RESULTS: Icotinib significantly reduced the right ventricular systolic pressure and right ventricle hypertrophy index in rats with MCT-induced PH. Moreover, icotinib improved MCT-induced pulmonary vascular remodeling. The expression of contractile marker (smooth muscle 22 alpha (SM22α)) and synthetic markers (osteopontin (OPN) and vimentin) in pulmonary artery was restored by icotinib treatment. In vitro, icotinib suppressed EGF-induced PASMCs proliferation and migration. Meanwhile, icotinib inhibited EGF-induced downregulation of α-smooth muscle actin and SM22α and upregulation of OPN and Collagen I in PASMCs, suggesting that icotinib could inhibit EGF-induced phenotypic switching of PASMCs. Mechanistically, these effects of icotinib were associated with the inhibition of EGFR-Akt/ERK signaling pathway. CONCLUSIONS: Icotinib can attenuate MCT-induced pulmonary vascular remodeling and improve PH. This effect of icotinib might be attributed to preventing PASMC dysfunction by inhibiting EGFR-Akt/ERK signaling pathway.

Dynamin-related protein-1 promotes lung cancer A549 cells apoptosis through the F-actin/bax signaling pathway.

Dynamin-related protein-1 (Drp1) has been found to be associated with cell death. The role of Drp1 in A549 cells death has not been explored. In this study, adenovirus-mediated Drp1 overexpression was used to investigate the influence of Drp1 on A549 cell viability with a focus on F-actin and Bax. Cell viability, protein expression, oxygen consumption, energy metabolism, and growth rate were measured through ELISA, qPCR, western blots and pathway analysis. Our results indicated that Drp1 overexpression promoted A549 cell death through apoptosis. Mechanistically, cytoskeletal F-actin was impaired and Bax expression was elevated in response to Drp1 overexpression. Besides, energy metabolism was reduced and oxygen consumption was interrupted. Therefore, our results demonstrated that A549 cell viability, apoptosis and growth were regulated by the Drp1/F-actin/Bax signaling pathways. These data explain a new role played by Drp1 in regulating cell viability and also provide a potential target to affect the progression of lung cancer through induction of cell death.

MARCKSL1 promotes the proliferation, migration and invasion of lung adenocarcinoma cells.

Lung cancer is the most common cancer in males and females and ~40% of lung cancer cases are adenocarcinomas. Previous studies have demonstrated that myristoylated alanine rich protein kinase C substrate (MARCKS) is upregulated in several types of cancer and is associated with poor prognosis in patients with breast cancer. However, its expression level and role in lung adenocarcinoma remain unknown. Therefore, the aim of the present study was to investigate the expression level and biological functions of MARCKS like 1 (MARCKSL1), a member of the MARCKS family, in lung adenocarcinoma. The expression level of MARCKSL1 was examined in human lung adenocarcinoma tissues and cell lines. MARCKSL1-specific small interfering RNAs effectively suppressed its expression level and significantly inhibited the proliferation, migration and invasion of lung adenocarcinoma cells. Additionally, the role of MARCKSLI in the regulation of metastasis was examined. Silencing MARCKSL1 decreased the expression of the epithelial-mesenchymal transition (EMT)-associated proteins E-cadherin, N-cadherin, vimentin and snail family transcriptional repressor 2, and decreased the phosphorylation level of AKT. The results obtained in the current study suggested that MARCKSL1 promoted the progression of lung adenocarcinoma by regulating EMT. MARCKSLI may have prognostic value and serve as a novel therapeutic target in lung adenocarcinoma.

Botulinum Toxin to Treat Horizontal Forehead Lines: A Refined Injection Pattern Accommodating the Lower Frontalis.

BACKGROUND: When treating horizontal forehead lines with botulinum toxin type A the traditional approach requires that injection points should stay 1.5 to 2 cm above the orbital rim to avoid brow ptosis. Failure to treat the lower frontalis may potentially cause worse rhytides in the lower forehead. OBJECTIVES: The aim of this study was to present a refined injection pattern accommodating the lower frontalis and evaluate its clinical efficacy and safety. METHODS: Patients were categorized into 4 types according to the patterns of their forehead wrinkles. Moderate and severe wrinkles in the upper forehead were treated by the "safe zone" technique. Mild wrinkles and rhytides in the lower forehead were treated by the Microbotox technique. Standard photographs and measurements were taken before and after treatment. The effect on wrinkle reduction and changes in brow heights were assessed. RESULTS: In total, 330 treatments were followed up in the clinic, and 246 treatments were followed up by telephone. Among the 330 treatments, 213 were evaluated in our clinic 2 to 4 weeks later, and the patients who received these treatments were recruited for effect evaluation and brow height measurements. The posttreatment severity of forehead wrinkles was significantly reduced (P < 0.05), and brow heights remained unchanged (P > 0.05). No severe adverse events were documented. Patient satisfaction was quite high. CONCLUSIONS: The refined injection pattern is an effective and safe technique to treat horizontal forehead lines. The Microbotox technique enables treatment of the lower frontalis without changes in brow position.

DNA methylation profiling analysis identifies a DNA methylation signature for predicting prognosis and recurrence of lung adenocarcinoma.

The aim of the present study was to identify candidate prognostic DNA methylation biomarkers for lung adenocarcinoma (LUAD), since the modern precise medicine for the treatment of LUAD requires more biomarkers and novel therapeutic targets of interest. DNA methylation profiling data of LUAD were downloaded from The Cancer Gene Atlas portal. Differentially methylated genes (DMGs) were screened to differentiate between samples designated as good and bad prognosis. LUAD-associated methylation modules were obtained with the weighted correlation network analysis (WGCNA) package, followed by function enrichment analysis. Optimal prognostic DMGs were selected using the LASSO estimation-based Cox-PH approach and were used to construct a prognostic risk scoring system. The training set was dichotomized by risk score, into high- and low-risk groups. The differences in overall survival (OS) time or recurrence-free survival (RFS) time between the two groups were evaluated using a Kaplan-Meier curve. A total of 742 DMG samples were screened for good and bad prognosis. WGCNA identified three LUAD-associated modules, which were primarily associated with cytoskeleton organization, transcription and apoptosis. A nine-gene prognostic methylation signature was determined, which included C20orf56, BTG2, C13orf16, DNASE1L1, ZDHHC3, FHDC1, ARF6, ITGB3 and ICAM4. A risk score-based methylation signature classified the patients in the training set into high- and low-risk groups with significantly different OS or RFS times. The prognostic value of the methylation signature was successfully verified in a validation set. In conclusion, the present study identified a nine-gene methylation signature for the prediction of survival and recurrence in patients with LUAD and improved the understanding of the alterations in DNA methylation in LUAD.

Plasma LUNX mRNA, a non-invasive specific biomarker for diagnosis and prognostic prediction of non-small cell lung cancer.

Lung cancer is the most common cancer worldwide. However, no specific biomarker has been found in diagnosis and evaluation of therapeutic efficacy for lung cancer. The human lung-specific X protein gene (LUNX) was recently identified with a feature of lung tissue specificity. We applied the fluorescent quantitative polymerase chain reaction method to examine LUNX mRNA in plasma and peripheral blood mononuclear cells (PBMC) in patients with non-small cell lung cancer (NSCLC), benign lung diseases, extrapulmonary tumors, and healthy subjects. The results showed that LUNX mRNA in both of plasma and PBMC were significantly higher in lung cancer patients compared to other groups. In plasma, there were higher sensitivity and negative predictive value of LUNX mRNA than in PBMC. Patients with III~IV stages of lung cancer had more LUNX mRNA in plasma than the early stage of lung cancer sufferers. After a period of therapy, significant reductions of plasma LUNX mRNA in patients with I and II stages of lung cancer were found. Levels of plasma LUNX mRNA in patients who had succeeded to respond to therapy decreased compared to prior treatment. On the other hand, the post-treatment level was obviously increased in patients that had failed to respond to therapy. Patients with negative plasma LUNX mRNA after therapy displayed a favorable prognosis and survival rate. These preliminary data suggested that cell-free LUNX mRNA in plasma as a non-invasive biomarker, is superior to peripheral intracellular LUNX mRNA, and plays a critical role in specific diagnosis and prognostic prediction of non-small cell lung cancer.

Discovery and validation of potential bacterial biomarkers for lung cancer.

Microbes are residents in a number of body sites, including the oral and nasal cavities, which are connected to the lung via the pharynx. The associations between oral diseases and increased risk of lung cancer have been reported in previous prospective studies. In this study, we measured variations of salivary microbiota and evaluated their potential association with lung cancer, including squamous cell carcinoma (SCC) and adenocarcinoma (AC). A three-phase study was performed: First, we investigated the salivary microbiota from 20 lung cancer patients (10 SCC and 10 AC) and control subjects (n=10) using a deep sequencing analysis. Salivary Capnocytophaga, Selenomonas, Veillonella and Neisseria were found to be significantly altered in patients with SCC and AC when compared to that in control subjects. Second, we confirmed the significant changes of Capnocytophaga, Veillonella and Neisseria in the same lung cancer patients using quantitative PCR (qPCR). Finally, these bacterial species were further validated on new patient/control cohorts (n=56) with qPCR. The combination of two bacterial biomarkers, Capnocytophaga and Veillonella, yielded a receiver operating characteristic (ROC) value of 0.86 with an 84.6% sensitivity and 86.7% specificity in distinguishing patients with SCC from control subjects and a ROC value of 0.80 with a 78.6% sensitivity and 80.0% specificity in distinguishing patients with AC from control subjects. In conclusion, we have for the first time demonstrated the association of saliva microbiota with lung cancer. Particularly, the combination of the 16S sequencing discovery with qPCR validation studies revealed that the levels of Capnocytophaga and Veillonella were significantly higher in the saliva from lung cancer patients, which may serve as potential biomarkers for the disease detection/classification.

Characterization of a Type 1 Metallothionein Gene from the Stresses-Tolerant Plant Ziziphus jujuba.

Plant metallothioneins (MTs) are a family of low molecular weight, cysteine-rich, and metal-binding proteins, which play an important role in the detoxification of heavy metal ions, osmotic stresses, and hormone treatment. Sequence analysis revealed that the open-reading frame (ORF) of ZjMT was 225 bp, which encodes a protein composed of 75 amino acid residues with a calculated molecular mass of 7.376 kDa and a predicated isoelectric point (pI) of 4.83. ZjMT belongs to the type I MT, which consists of two highly conserved cysteine-rich terminal domains linked by a cysteine free region. Our studies showed that ZjMT was primarily localized in the cytoplasm and the nucleus of cells and ZjMT expression was up-regulated by NaCl, CdCl2 and polyethylene glycol (PEG) treatments. Constitutive expression of ZjMT in wild type Arabidopsis plants enhanced their tolerance to NaCl stress during the germination stage. Compared with the wild type, transgenic plants accumulate more Cd2+ in root, but less in leaf, suggesting that ZjMT may have a function in Cd2+ retension in roots and, therefore, decrease the toxicity of Cd2+.

Interaction of ribosomal protein L22 with casein kinase 2α: a novel mechanism for understanding the biology of non-small cell lung cancer.

Dysfunction of ribosomal proteins (RPs) may play an important role in molecular tumorigenesis, such as lung cancer, acting in extraribosomal functions. Many protein-protein interaction studies and genetic screens have confirmed the extraribosomal capacity of RPs. As reported, ribosomal protein L22 (RPL22) dysfunction could increase cancer risk. In the present study, we examined RPL22-protein complexes in lung cancer cells. Tandem affinity purification (TAP) was used to screen the RPL22-protein complexes, and GST pull-down experiments and confocal microscopy were used to assess the protein-protein interaction. The experiment of kinase assay was used to study the function of the RPL22-protein complexes. The results showed that several differentially expressed proteins were isolated and identified by LC-MS/MS, which revealed that one of the protein complexes included casein kinase 2α (CK2α). RPL22 and CK2α interact in vitro. RPL22 also inhibited CK2α substrate phosphorylation in vitro. This is the first report of the RPL22-CK2α relationship in lung cancer. Dysregulated CK2 may impact cell proliferation and apoptosis, key features of cancer cell biology. Our results indicate that RPL22 may be a candidate anticancer agent due to its CK2α-binding and -inhibitory functions in human lung cancer.

Down-regulation of ribosomal protein L22 in non-small cell lung cancer.

Ribosomal protein L22 (RPL22), an RNA-binding protein, is a constituent of the 60S large ribosomal subunit. As reported, RPL22 is not required in protein synthesis, and mutations of RPL22 were the main cause of macrolide resistance in bacteria. In vertebrates, RPL22 mutation might increase the proliferation of cells and then increase cancer risk. However, to our knowledge, RPL22 has not been implicated in any lung diseases, especially in lung cancer. In this study, we compared the expression of RPL22 gene in non-small cell lung cancer (NSCLC) tissues, plasma as well as human lung cancer cell line LTEP-a-2 with that in normal lung tissues and cells, using real-time RT-qPCR, Western blot, quantitative immunohistochemistry analysis, and ELISA. Our studies showed that the expression of RPL22 was significantly down-regulated in mRNA and protein expression level in NSCLC; however, there was no significant difference of RPL22 levels in plasma between normal and NSCLC patients. Further analysis indicated that down-regulation of RPL22 might be involved in the carcinogenesis of NSCLC, yet not an effective biomarker in plasma for early diagnosis.