Inhibition of Hypothalamic FTO Activates STAT3 Signal through ERK1/2 Associated with Reductions in Food Intake and Body Weight.

INTRODUCTION: Fat mass and obesity-associated (FTO) gene is strongly associated with obesity which brings a major health threat. Altered expression of its encoded protein FTO in the hypothalamus has been identified to contribute to central control of appetite and body weight. However, its molecular mechanisms remain elusive. METHODS: Mouse hypothalamic POMC cell line N43/5 was treated with FTO inhibitor rhein, FTO shRNA, or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor U0126 to inhibit FTO or ERK1/2. Rhein and U0126 were injected into lateral ventricle of the mice by intracerebroventricular cannulation. Western blotting and immunofluorescent assays were performed to monitor protein level. RESULTS: This study identified that inhibition of FTO in N43/5 cells led to phosphorylation of signal transducer and activator of transcription 3 (STAT3) at S727 site and induced p-STAT3-S727 nuclear translocation. We further showed that FTO inhibition promoted phosphorylation of ERK1/2; specific inhibition of ERK1/2 signaling by U0126 could abolish the effect of FTO inhibition on STAT3-S727 phosphorylation and nuclear translocation. Furthermore, we found that inhibition of hypothalamic FTO promoted STAT3-S727 phosphorylation in the hypothalamic arcuate nucleus, and the mice showed reductions in food intake and body weight. In addition, inhibition of hypothalamic ERK1/2 could abolish the effects of FTO inhibition on STAT3-S727 phosphorylation, reductions of food intake and body weight. CONCLUSION: Our in vitro and in vivo data suggest that the inhibition of hypothalamic FTO could activate STAT3 through ERK1/2, which is potentially associated with reductions in food intake and body weight.

Symmetry Breaking and Anomalous Conductivity in a Double-Moiré Superlattice.

In a two-dimensional moiré superlattice, the atomic reconstruction of constituent layers could introduce significant modifications to the lattice symmetry and electronic structure at small twist angles. Here, we employ conductive atomic force microscopy to investigate a twisted trilayer graphene double-moiré superlattice. Two sets of moiré superlattices are observed. At neighboring domains of the large moiré, the current exhibits either 2- or 6-fold rotational symmetry, indicating delicate symmetry breaking beyond the rigid model. Moreover, an anomalous current appears at the "A-A" stacking site of the larger moiré, contradictory to previous observations on twisted bilayer graphene. Both behaviors can be understood by atomic reconstruction, and we also show that the measured current is dominated by the tip-graphene contact resistance that maps the local work function qualitatively. Our results reveal new insights of atomic reconstruction in novel moiré superlattices and opportunities for manipulating exotic quantum states on the basis of twisted van der Waals heterostructures.

The Features and Progress of Electrolyte for Potassium Ion Batteries.

Nowadays, Li-ion batteries have achieved great success and are widely used in various fields. However, the scarcity and uneven distribution of lithium resources together with the increasing cost may hamper the sustainable development of Li-ion batteries in the future. Hence, many researchers have turned to potassium ion batteries due to their abundant raw materials, low price, and high energy density. Although great progress has been made in recent years, there are still existing many challenges, especially the severe side reaction between electrolyte and K metal, which leads to an unstable solid-liquid interface and low coulombic efficiency. Hence, an excellent electrolyte may be the key to development of K-ion batteries in the future. Unfortunately, no systematic research has been conducted to study the electrolyte and its role on the performance yet. In order to compensate for this limitation, in this paper, the status and progress of electrolytes for K-ion batteries are reviewed, the issues and challenges existing in the development of electrolyte are clarified, and the future development is prospected.

Elastic properties and intrinsic strength of two-dimensional InSe flakes.

The mechanical properties of two-dimensional (2D) materials are critical for their applications in functional devices as well as for strain engineering. Here, we report the Young's modulus and breaking strength of multilayered InSe, an emerging 2D semiconductor of the layered group III chalcogenide. Few-layer InSe flaks were exfoliated from bulk InSe crystal onto Si/SiO2 substrate with micro-fabricated holes, and indentation tests were carried out using an atomic force microscopy probe. In combination with both continuum analysis and finite element simulation, we measured the Young's modulus of multilayer 2D InSe (>5 L) to be 101.37 ± 17.93 GPa, much higher than its bulk counterpart, while its breaking strength is determined to be 8.68 GPa, approaching the theoretical limit of 10.1 GPa. Density functional theory calculations were also carried out to explain the insensitivity of Young's modulus to the layer count. It is found that 2D InSe is softer than most 2D materials, and exhibits breaking strength higher than that of carbon fiber, yet remaining more compliant, making it ideal for flexible electronics applications. The reliability of our method is also validated by measurement of graphene.

Neuroprotective Effect of Modified Electroconvulsive Therapy for Schizophrenia: A Proton Magnetic Resonance Spectroscopy Study.

The underlying mechanism of modified electroconvulsive therapy (MECT) treatment for drug-resistant and catatonic schizophrenia remains unclear. Here, we aim to investigate whether MECT exerts its antipsychotic effects through elevating N-acetylaspartate (NAA) concentration measured by proton magnetic resonance spectroscopy (H-MRS). Multiple-voxel H-MRS was acquired in the bilateral prefrontal cortex (PFC) and thalamus to obtain measures of neurochemistry in 32 MECT, 34 atypical antipsychotic-treated schizophrenic patients, and 34 healthy controls. We found that both MECT and atypical antipsychotic treatments showed significant antipsychotic efficacy. MECT and atypical antipsychotic treatments reversed the reduced NAA/creatine ratio (NAA/Cr) in the left PFC and left thalamus in schizophrenic patients compared with healthy controls. Furthermore, the NAA/Cr ratio after treatments was significant higher in the MECT group, but not in the medication group. Our findings demonstrate that eight times of MECT elevated the relative NAA concentration to display neuroprotective effect, which may be the underlying mechanism of rapid antipsychotic efficacy.

Regulator of G-protein signaling 4: A novel tumor suppressor with prognostic significance in non-small cell lung cancer.

Regulator of G-protein signaling (RGS) family members are regulatory molecules which act as GTPase activating proteins (GAPs) for G alpha subunits of heterotrimeric G proteins. Emerging data indicated that RGS members were involved with tumorigenesis and metastasis. In the current study, we identified RGS4 as a novel tumor suppressor with prognostic significance in non-small cell lung cancer (NSCLC). To be specific, we found that RGS4 expression was higher in normal lung tissues than NSCLC specimens (P = 0.003). Further studies demonstrated that RGS4 was generally down-regulated in NSCLC specimens compared with the matched normal lung tissues, both at mRNA and protein levels. In addition, correlational analysis indicated that RGS4 expression levels negatively correlated with lymph node metastasis (P = 0.009) and TNM stage (P = 0.008). Survival analysis demonstrated that patients with lower RGS4 protein expression exhibited a much worse 5-year overall survival and 5-year disease-free survival than those with high expression. More importantly, we proved that over-expression of RGS4 in NSCLC cells decreased invasion and migration due to inhibition of MMP2/9 and reversal of EMT while down-regulation of RGS4 in normal lung cell lines promoted invasion and migration. At last, nude mice metastatic model proved that over-expression of RGS4 suppressed tumor metastasis in vivo. All of these results confirmed the critical role of RGS4 in NSCLC progression.

Rotenone induces apoptosis in human lung cancer cells by regulating autophagic flux.

Reactive oxygen species (ROS) are at the center of many physiological and pathological processes. ROS generated due to oxidative stress can potentiate both cancer initiation and progression. Rotenone, which is an inhibitor of the mitochondrial electron transport chain complex I, results in the activation of NOX2 and release of ROS, and has been shown to display anticancer activity through the induction of apoptosis in various cancer cells. The mechanistic link between rotenone-dependent activation of NOX2 and induction of apoptosis is still elusive. In this study, we used the human lung cancer A549 cells to study the molecular mechanism(s) involved between rotenone-dependent activation of NOX2 and impairment of autophagic machinery. We report that acute exposure to rotenone induced mild NOX2-dependnet oxidative stress, which impaired the autophagic flux, resulting in cytosolic accumulation of LC3 and p62/STSQM1. We further show that this induction occurs through the PI3K/Akt/mTORC1 signaling pathway. We furthermore show that chronic exposure to rotenone lead to excessive NOX2-dependent ROS generation, increases autophagy, and decreases p62 level via increased-autophagic flux. Taken together, this study is the first mechanistic elucidation of how rotenone can be used to potently target cancer cells without overhauling the entire cellular machinery. © 2016 IUBMB Life 68(5):388-393, 2016.

Hippocampal proteomic changes in high-fat diet-induced obese mice associated with memory decline.

Substantial evidence suggest that chronic consumption of high-fat diets (HFDs) can lead to obesity, abnormal metabolism, as well as cognitive impairment. Molecular and cellular changes regarding hippocampal dysfunctions have been identified in multiple HFD animal models. Therefore, in-depth identification of expression changes of hippocampal proteins is critical for understanding the mechanism of HFD-induced cognitive deficits. In this study, we fed 3-week-old male mice with HFD for 3 months to generate obese mice who exhibit systemic metabolic abnormality and learning and memory decline. Using an iTRAQ-labeled proteomic analysis, we identified a total of 82 differentially expressed proteins (DEPs) in the hippocampus upon HFD with 35 up-regulated proteins and 47 down-regulated proteins. Functional enrichment indicated that these DEPs were predominantly enriched in regulation of catabolic process, dendritic shaft, neuron projection morphogenesis and GTPase regulator activity. Protein-protein interaction enrichment showed that the DEPs are mostly enriched in postsynaptic functions; and of them, six proteins (i.e., DLG3, SYNGAP1, DCLK1, GRIA4, GRIP1, and ARHGAP32) were involved in several functional assemblies of the postsynaptic density including G-protein signaling, scaffolding and adaptor, kinase and AMPA signaling, respectively. Collectively, our findings suggest that these DEPs upon HFD might contribute to memory decline by disturbing neuronal and postsynaptic functions in the hippocampus.

Down-regulation of MTA1 protein leads to the inhibition of migration, invasion, and angiogenesis of non-small-cell lung cancer cell line.

Metastasis-associated protein 1 (MTA1) high expression has been detected in a wide variety of human aggressive tumors and plays important roles in the malignant biological behaviors such as invasion, metastasis, and angiogenesis. However, the specific roles and mechanisms of MTA1 protein in regulating the malignant behaviors of non-small-cell lung cancer (NSCLC) cells still remain unclear. To elucidate the detailed functions of MTA1 protein, we down-regulated the MTA1 protein expression in NSCLC cell line by RNA interference (RNAi) in vitro, and found that down-regulation of MTA1 protein significantly inhibited the migration and invasion potentials of 95D cells. Further research revealed that down-regulation of MTA1 protein significantly decreased the activity of matrix metalloproteinase-9, which could be the mechanism responsible for the inhibition of 95D cells migration and invasion. In addition, the tube formation assay demonstrated that the number of complete tubes induced by the conditioned medium of MTA1-siRNA 95D cells was significantly smaller than that of 95D cells. These findings demonstrate that MTA1 protein plays important roles in regulating the migration, invasion, and angiogenesis potentials of 95D cells, suggesting that MTA1 protein down-regulation by RNAi might be a novel therapeutic approach to inhibit the progression of NSCLC.

Metastasis-associated protein 1 nuclear expression is closely associated with tumor progression and angiogenesis in patients with esophageal squamous cell cancer.

BACKGROUND: The purposes of the present study were to detect the expression of metastasis-associated protein 1 (MTA1) in patients with esophageal squamous cell cancer (ESCC), and to evaluate the relevance of MTA1 protein expression to the tumor progression, angiogenesis, and prognosis. METHODS: Both MTA1 protein and intratumoral microvessels were examined by immunohistochemical staining in 131 ESCC patients who successfully underwent subtotal esophagectomy and esophagogastric anastomosis at Qilu Hospital between Jan 2004 and Dec 2005. Intratumoral microvessel density (MVD) was recorded by counting CD-34 positive immunostained endothelial cells. All statistical analyses were performed with SPSS 13.0 statistical software. RESULTS: High expression of MTA1 protein was detected in 57 cases and significantly correlated with tumor invasion depth (P = 0.041), lymph node metastasis (P = 0.021), pathologic stage (P = 0.003), and MVD (P = 0.044). Survival analysis showed that patients with MTA1 protein high expression had significantly poor overall 5-year survival (P = 0.002), and the factor found on multivariate analysis to significantly affect overall survival was only pathologic stage (P = 0.040). Further stratified survival analysis split by pathologic stage demonstrated that MTA1 protein high expression significantly predicted unfavorable prognosis among patients with pathologic stage II disease (P = 0.006). CONCLUSIONS: High expression of the MTA1 protein is common in ESCC, and is closely associated with tumor progression, increased tumor angiogenesis, and poor survival. These findings indicate that MTA1 protein has clinical potentials as a useful indicator of progressive phenotype, a promising prognostic predictor to identify patients with poor prognosis, and a potential novel therapeutic target of antiangiogenesis for patients with ESCC.

Diallyl trisulfide induces apoptosis and inhibits proliferation of A549 cells in vitro and in vivo.

Lung cancer is the leading cause of cancer-related mortality all over the world. In recent years, pulmonary adenocarcinoma has surpassed squamous cell carcinoma in frequency and is the predominant form of lung cancer in many countries. Epidemiological investigations have shown an inverse relationship between garlic (Allium sativum) consumption and death rate from many cancers. Diallyl trisulfide (DATS) is one of the garlic-derived compounds (also known as: organosulfer compounds, OSC). DATS can induce apoptosis and inhibit the growth of many cancer cell lines. Our study demonstrated that the apoptotic incidents induced by DATS were a mitochondria-dependent caspase cascade through a significant decrease of the anti-apoptotic Bcl-2 that resulted in up-regulation of the ratio of Bax/Bcl-2 and the activity of caspase-3, -8, and -9. Eventually, DATS induced the apoptosis and inhibited the proliferation in a concentration- and time-dependent manner. Furthermore, by establishing an animal model of female BALB/c nude mice with A549 xenografts, we found that oral gavage of DATS significantly retarded growth of A549 xenografts in nude mice without causing weight loss or any other side effects compared with the control group. All the evidence both in vitro and in vivo suggested that DATS could be an ideal anti-cancer drug.

[Proton magnetic resonance spectroscopy study of prefrontal lobe and thalamus in schizophrenics on modified electroconvulsive therapy].

OBJECTIVE: To explore the potential effects of modified electroconvulsive therapy (MECT) in prefrontal lobe and thalamus in patients with schizophrenia by proton magnetic resonance spectroscopy ((1)H-MRS). METHODS: From November 2010 to June 2011, a total of 31 schizophrenics fulfilling the third edition of the Chinese Classification of Mental Disorders (CCMD-III) were recruited. And prefrontal lobe and thalamus were evaluated by multi-voxel (1)H-MRS before and after 8 sessions of MECT. The subjects were evaluated by the positive and negative syndrome scale (PANSS). And the N-acetylaspartate (NAA), choline-containing compounds (Cho) and creatine compounds (Cr) were measured and the ratios of NAA/Cr and Cho/Cr determined. RESULTS: (1) In left prefrontal lobe and bilateral thalamus, the NAA/Cr ratio at post-treatment demonstrated higher than that at pre-treatment (1.50 ± 0.31 vs 1.35 ± 0.30, t = 2.07, P < 0.05; 1.53 ± 0.31 vs 1.38 ± 0.27, t = 2.03, P < 0.05; 1.51 ± 0.29 vs 1.36 ± 0.26, t = 2.14, P < 0.05). (2) The major influencing factors of the changes of NAA/Cr in left prefrontal lobe were age of onset, decrease rate of PANSS, baseline PANSS total score and duration of illness. And the major influencing factors for left thalamus were age of onset and duration of illness while a major influencing factor for right thalamus was baseline PANSS total score. CONCLUSION: MECT may modify brain metabolism as measured by (1)H-MRS. The pattern of changes suggests possible neuroprotective effects in schizophrenics. And these effects are correlated with age of onset, duration and severity of illness.

Corrigendum: Negatively regulated by miR-29c-3p, MTFR1 promotes the progression and glycolysis in lung adenocarcinoma via the AMPK/mTOR signalling pathway.

[This corrects the article DOI: 10.3389/fcell.2021.771824.].

[Proton magnetic resonance spectroscopy of prefrontal lobe and thalamus in schizophrenics: correlation with clinical characteristics].

OBJECTIVE: To explore the characteristics of different subtypes of schizophrenics on prefrontal lobe and thalamus by proton magnetic resonance spectroscopy ((1)H-MRS) and its relationship. METHODS: From August 2007 to April 2010 at our center, a total of 159 schizophrenics fulfilling the third edition criteria of Chinese Classification of Mental Disorders (CCMD-III) were recruited. And prefrontal lobe and thalamus were evaluated by multi-voxel (1)H-MRS. There were 88 males and 71 females. There were first-episode (n = 54) and not-first-episode (n = 105), negative subtype (n = 125) and positive subtype (n = 34), medicated (n = 96) and non-medicated (n = 63) by different criteria. The levels of N-acetylaspartate (NAA), choline-containing compounds (Cho) and creatine compounds (Cr) were measured and the ratios of NAA/Cr and Cho/Cr determined. Positive and negative syndrome scale (PANSS) and Wisconsin card sorting test (WCST) were also assessed. Only 45 normal controls received (1)H-MRS. RESULTS: On left prefrontal lobe and left thalamus, the NAA/Cr ratios in different subtypes of patients were lower than those in normal controls (P < 0.05 or 0.01). The NAA/Cr ratios in patients of non-first-episode (1.48 ± 0.34), negative subtype (1.40 ± 0.35) and medicated (1.47 ± 0.36) on right thalamus were also lower than those in normal controls (1.62 ± 0.37, t = 2.25, 3.56, 2.28, P < 0.05 or P < 0.01). Compared with positive subtype schizophrenics, the NAA/Cr ratios in those of negative subtype on right thalamus were lower (1.40 ± 0.35 vs 1.60 ± 0.37, t = 2.92, P < 0.01). On right thalamus of non-medicated schizophrenics, there was a negative correlation between the duration of illness and the ratio of NAA/Cr (r = -0.38, P < 0.05) and a positive correlation between the duration of illness and the ratio of Cho/Cr (r = 0.43, P < 0.01). On right thalamus of negative subtype schizophrenics, the ratios of NAA/Cr were negatively correlated with the total score of PANSS and the score of negative factor respectively (r = -0.36, -0.40, P < 0.05). On left prefrontal lobe of different subtypes, the ratios of NAA/Cr were negatively correlated with the total score of PANSS, the score of negative factor, responses errors and persistent errors (P < 0.01) and positively correlated with completed categories and conceptual level responses (P < 0.05 or P < 0.01). CONCLUSION: Abnormalities in neuronal function and/or integrity are present on left prefrontal lobe and left thalamus in schizophrenics. And right thalamus is probably involved in non-first-episode subtype, negative subtype and non-medicated subtype. Different subtypes of schizophrenics may have different characteristics of (1)H-MRS due to the duration of illness and their clinical symptoms.

MCTS1 promotes the development of lung adenocarcinoma by regulating E2F1 expression.

Lung adenocarcinoma (LUAD) is the most common subtype of lung cancer that results in the majority of cancer-associated mortality. Multiple copies in T-cell lymphoma-1 (MCTS1) is an oncogene that is expressed at high levels in several types of cancer tissues. However, its exact role and pathomechanism in the development of LUAD remains unknown. Reverse transcription-quantitative PCR analysis was performed to detect MCTS1 expression. Immunohistochemistry analysis was performed to detect MCTS1 expression in LUAD tissues and normal tissues. The MTT, colony formation, EdU, flow cytometry, wound healing and Transwell assays were performed to assess the proliferation, apoptosis, migration and invasion of LUAD cells. Western blot analysis was performed to detect protein expression levels. The present study aimed to investigate the effects of MCTS1 on the progression of LUAD and the potential mechanisms underlying its effects. The results demonstrated that MCTS1 expression was upregulated in LUAD tissues and cells, which was associated with an unfavorable outcome in patients with LUAD. MCTS1 knockdown inhibited LUAD progression by suppressing cell viability and motility, and promoting apoptosis. In addition, E2F1 protein expression was attenuated following MCTS1 knockdown. The silencing MCTS1-induced inhibitory effect on LUAD malignancy was reversed following overexpression of E2F1 by modulating the c-Myc signaling pathway. Taken together, the results of the present study suggest that MCTS1 facilitates cell proliferation and migration, and suppresses apoptosis of LUAD cells by regulating E2F1 expression and the c-Myc signaling pathway.

Negatively Regulated by miR-29c-3p, MTFR1 Promotes the Progression and Glycolysis in Lung Adenocarcinoma via the AMPK/mTOR Signalling Pathway.

Background: Lung adenocarcinoma (LUAD) is the major form of lung cancer that presents a major peril to public health. Owing to the high rates of morbidity, mortality and chemoresistance, it is necessary to develop more effective therapeutic targets of LUAD. Mitochondrial fission regulator 1 (MTFR1) affects the occurrence and development of some diseases by regulating mitochondrial dynamics and is dysregulated in LUAD. However, the functions and molecular mechanisms of MTFR1 in LUAD have not been investigated. Methods: Immunohistochemical (IHC) analysis, real-time quantitative polymerase chain reaction (RT-qPCR), bioinformatic analysis and western blot (WB) were performed to assess the expression of MTFR1 at both protein and mRNA levels. The biological functions of MTFR1 in LUAD cells were assessed based on various in vivo and in vitro experiments. The dual-luciferase reporter assay and some rescue experiments were performed to evaluate the underlying mechanism of MTFR1 in LUAD. Results: MTFR1 was upregulated in LUAD cells and tissues and correlated with dismal clinicopathologic features and a worse prognosis of patients with LUAD. Functionally, MTFR1 overexpression stimulated the proliferation, invasion, migration and glycolytic capacity and impeded the apoptosis of LUAD cells; however, opposite results were obtained when MTFR1 expression was knocked down. MTFR1, which was directly targeted by miR-29c-3p, may exert its biological functions through the AMPK/mTOR signalling pathway. Conclusion: MTFR1 promotes the progression of LUAD. Therefore, targeting MTFR1 can offer an effective therapeutic strategy for LUAD treatment.

Circular RNA hsa-circ-000881 suppresses the progression of lung adenocarcinoma in vitro via a miR-665/PRICKLE2 axis.

BACKGROUND: Circular RNA (circRNA) has become a new focus in the field of tumor biology research in recent years. Many circRNAs have been showed to play an important role in the progression of lung adenocarcinoma (LUAD). In this work, we studied the oncological role of hsa-circ-000881 in LUAD and attempted to explore the related mechanism. METHODS: The relative expressions of hsa-circ-000881, miR-665, and PRICKLE2 were detected by RT-qPCR or western blot. Functional assays were conducted to analyze the role of hsa-circ-000881 in the proliferation, migration, and invasion of LUAD cells. A luciferase reporter assay was performed to verify whether hsa-circ-000881, miR-665, and PRICKLE2 interact with each other. RESULTS: Circ-000881 was remarkably downregulated in LUAD. Overexpression of circ-000881 attenuated cell growth, migration, and invasion, whereas its knockdown enhanced the malignancy of LUAD cells. The results of luciferase reporter assay and bioinformatics analysis confirmed that circ-000881 served as a sponge for miR-665, and PRICKLE2 was a direct target of miR-665.Overexpression of miR-665 or silencing of PRICKLE2 abolished circ-000881-mediated inhibition of malignant tumor behavior in LUAD cells. CONCLUSIONS: Circ-000881 has inhibitory effects on LUAD via a miR-665/PRICKLE2 axis, suggesting that circ-000881 may be an underlying therapeutic target for LUAD.

Expression of MiR-608 in Nonsmall Cell Lung Cancer and Molecular Mechanism of Apoptosis and Migration of A549 Cells.

OBJECTIVE: This Work is aimed at exploring the effect of microRNA (MiR)-608 on the function of nonsmall cell lung cancer (NSCLC) A549 cells and related mechanisms. METHODS: Blood samples of 106 NSCLC patients (experimental group) as well as 124 normal people (control group) were selected for relevant investigation. Polymerase chain reaction (PCR) as well as DNA sequencing was used to determine the genotyping of the MiR-608 rs4919510 polymorphism. MiR-608 expression in cells was detected by real-time PCR technology. Western blotting was used to detect changes in protein levels. NSCLC tissues as well as adjacent tissues were explored in 33 patients undergoing surgery. RESULTS: MiR-608 rs4919510 does not influence the incidence of NSCLC patients. In addition, MiR-608 expression was downregulated in the tumor tissue of NSCLC patients, while the transcription factor activating enhancer-binding protein 4 (TFAP4) expression was upregulated. MiR-608 promotes DOX- (Doxorubicin-) induced apoptosis by negatively regulating TFAP4 expression in NSCLC tissue. TFAP4 can significantly inhibit the migration of A549 cells. CONCLUSION: The findings in this investigation can contribute to the effective treatment of NSCLC patients. Also, the investigation can provide some theoretical support for the application of new targets for NSCLC treatment.

Correlation of plasma exosomal microRNAs with the efficacy of immunotherapy in EGFR/ALK wild-type advanced non-small cell lung cancer.

BACKGROUND: Immunotherapy has become an important treatment option for patients with advanced non-small cell lung cancer (NSCLC). At present, none of these existing biomarkers can effectively stratify true responders and there is an urgent need for identifying novel biomarkers. Exosomes derived from the serum of patients with cancer have been proven to be reliable markers for cancer diagnosis. Here, we explored the possibility of using plasma-derived exosomal microRNAs as potential biomarkers for optimal selection of patients with advanced EGFR/ALK negative NSCLC to immunotherapy. METHODS: From June 2017 to February 2019, 30 patients with advanced EGFR/ALK wild-type (WT) NSCLC who received PD-1/PD-L1 inhibitors were enrolled. The efficacy evaluation was conducted after every three cycles of treatment according to RECIST 1.1. Plasma samples of these patients were collected before the administration of PD-1/PD-L1 inhibitors as baseline, and after every three cycles if the patients achieved partial response (PR) or complete response. Plasma from seven healthy individuals was also collected as normal control. Exosomes were prepared by ultracentrifugation followed by total RNA extraction, and exosome-derived miRNAs were profiled using small RNA next-generation sequencing followed by differential expression analysis. RESULTS: In order to identify biomarker for better response, all five patients who achieved PR and four patients with progressive disease (PD) at efficacy evaluation were included for differential expression analysis. Based on unsupervised hierarchical clustering, exosomal miRNA expression profile was significantly altered in patients with NSCLC compared with normal controls with a total of 155 differentially expressed exosomal miRNAs. Interestingly, hsa-miR-320d, hsa-miR-320c, and hsa-miR-320b were identified significantly upregulated in the PD groups compared with the PR group at baseline before the treatment. In addition, we identified that hsa-miR-125b-5p, a T-cell suppressor, showed a trend of increased expression in the PD group at baseline and was significantly downregulated in the post-treatment plasma exosomes compared with pre-treatment samples of the PR patients. CONCLUSION: Patients with NSCLC represent unique plasma exosomal miRNA profiles. Hsa-miR-320d, hsa-miR-320c, and hsa-miR-320b were identified as potential biomarkers for predicting the efficacy of immunotherapy in advanced NSCLCs. When T-cell suppressor hsa-miR-125b-5p was downregulated during the treatment, the patients may obtain increased T-cell function and respond well to immunotherapy.

[Impacts of microparticles on vascular endothelial cells].

OBJECTIVE: To investigate the effects of microparticles (Mps) from different people on the vascular endothelial cell function. METHODS: Third generation human umbilical vein endothelial cells (HUVECs) were cultured with Mps-containing plasma samples from 5 systemic lupus erythematosus (SLE) patients undergoing heavy steroid treatment, 5 patients with steroid-induced avascular necrosis of femoral head (ANFH), 4 patients with alcohol induced ANFH, and 4 healthy persons for 12 h, 24 h, and 48 h respectively. Plasma samples from the above persons with the Mps filtered were used as experimental controls. HUVECs cultured with blank culture fluid were used as blank controls. Inverse phase contrast microscopy was used to observe the morphology of the HUVECs. PE-CD31 and PEcy5-CD62E were added into the flow cytometric test tubes and flow cytometry (FC) was used to count the number of CD62E +/ CD31 + Mps in the medium. RT-PCR was used to measure the mRNA expression of the apoptotic gene fas (fas/beta-actin). RESULTS: Microscopy showed no distinct difference between the morphology of the HUVECs among the different groups. FC showed that the number of CD62E +/CD31 + Mps 48 hours after the 20% Mp stimulation of the steroid-treated SLE group was significantly lower than that of the control group (P = 0.035). RT-PCR showed that 48 hours after the stimulation the levels of mRNA fas/beta-actin of the steroid-treated SLE group and steroid induced ANFH group were 0.914 +/- 0.226 and 0.776 +/- 0.230 respectively, both significantly higher than those of the control groups (0.832 +/- 0. 200 and 0.669 +/- 0.148 respectively, P = 0.005 and P = 0.006). CONCLUSION: The Mps from the steroid treated patients aggravate the apoptosis of HUVECs, and the Mps from the steroid induced ANFH patients augment the production of apoptosis gene in HUVECs. The Mps from healthy people and alcohol induced ANFH patients have no relationship with HUVEC apoptosis.