An EEG Channel Selection Framework for Driver Drowsiness Detection via Interpretability Guidance.

Drowsy driving has a crucial influence on driving safety, creating an urgent demand for driver drowsiness detection. Electroencephalogram (EEG) signal can accurately reflect the mental fatigue state and thus has been widely studied in drowsiness monitoring. However, the raw EEG data is inherently noisy and redundant, which is neglected by existing works that just use single-channel EEG data or full-head channel EEG data for model training, resulting in limited performance of driver drowsiness detection. In this paper, we are the first to propose an Interpretability-guided Channel Selection (ICS) framework for the driver drowsiness detection task. Specifically, we design a two-stage training strategy to progressively select the key contributing channels with the guidance of interpretability. We first train a teacher network in the first stage using full-head channel EEG data. Then we apply the class activation mapping (CAM) to the trained teacher model to highlight the high-contributing EEG channels and further propose a channel voting scheme to select the top N contributing EEG channels. Finally, we train a student network with the selected channels of EEG data in the second stage for driver drowsiness detection. Experiments are designed on a public dataset, and the results demonstrate that our method is highly applicable and can significantly improve the performance of cross-subject driver drowsiness detection.

Bifidobacterium breve predicts the efficacy of anti-PD-1 immunotherapy combined with chemotherapy in Chinese NSCLC patients.

BACKGROUND AND PURPOSE: Gut microbes play an important role in the occurrence of lung cancer, immunotherapy, and chemotherapy. In this study, we analyzed the characteristics of gut microbes in patients with lung cancer and investigated the effect of gut microbes on anti-PD-1 therapy combined with chemotherapy. METHODS: Fecal samples from 21 non-small cell lung cancer (NSCLC) patients and 22 healthy volunteers who were treated in the Fourth Hospital of Hebei Medical University from 2019 to 2021 were collected. DNA was extracted from all samples, and the V3-V4 region of the bacterial 16S rRNA gene was PCR-amplified using the Illumina sequencing platform, and R language was used for data analysis. RESULTS: There were significant differences in the Beta diversity and metabolic pathways of gut microbes between NSCLC patients and healthy individuals (p < 0.05). Bifidobacterium, Escherichia, and Sarterella were significantly enriched in patients with clinical benefit response (p < 0.05), and these three bacteria had certain predictive value for clinical benefit. Patients with Bifidobacterium breve had significantly longer median progression-free survival (mPFS) compared with patients with no detectable Bifidobacterium breve feces at baseline (106 days vs. NR, p < 0.001). Multivariate COX analysis showed that the presence of B.breve was an independent good prognostic factor affecting the PFS of patients receiving combination therapy (p < 0.05). CONCLUSION: The clinical efficacy of anti-PD-1 therapy combined with chemotherapy in Chinese advanced NSCLC patients is closely related to the gut microbiota, and Bifidobacterium breve may be a potential biomarker to predict the efficacy of immune-combined chemotherapy.

Effects of the vibrational stimulation on the muscle strength of the knee joints of athletes / Efeitos da estimulação da vibração sobre a força muscular das articulações do joelho dos atletas / Efectos de la estimulación por vibración en la fuerza muscular de las articulaciones de atletas

ABSTRACT Introduction: The traditional lower extremity muscle strength training consists mainly of resistance training, where training intensity is gradually increased, targeting strength gain. Objective: Study the effect of different vibration frequencies on muscle strength training of tennis players' knee joints. Methods: Using PHYSIO-PLATE vibration training platform, tennis players of a Beijing team were subjected to different frequencies of strength training with vibrational stimulation; after eight weeks of systematic strength training, the vibration frequencies were 30Hz and 45Hz, with amplitude of 7mm. Results: After the experiment, the relative peak torque and total work of the knee extensor muscles in subjects in groups I and II were significantly improved (P<0.05), generating a significant increase in rapid maximal power start. Conclusion: The vibrational stimulation addition to muscle strength training can effectively enhance its effect, including characteristics such as maximal strength, rapid strength, and muscular endurance with a relatively small load. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: O método tradicional de treinamento de força muscular das extremidades inferiores consiste principalmente no treinamento de resistência, onde é aumentada gradualmente a intensidade do treinamento visando o ganho de força. Objetivo: Estudar o efeito de diferentes frequências de vibração sobre o treino de força muscular nas articulações do joelho de tenistas. Métodos: Usando a plataforma de treinamento de vibração PHYSIO-PLATE, os tenistas de uma equipe de Pequim foram submetidos a diferentes frequências de treinamento de força com estimulação vibratória, após oito semanas de treinamento de força sistemático, as frequências de vibração foram de 30Hz e 45Hz, com amplitude de 7mm. Resultados: Após o experimento, o torque de pico relativo e o trabalho total dos músculos extensores do joelho nos indivíduos dos grupos I e II foram significativamente aprimorados (P<0,05), gerando um significativo aumento na potência máxima rápida de arranque. Conclusão: A adição do estímulo vibratório ao treinamento de força muscular pode efetivamente melhorar seu efeito, incluindo características como a força máxima, força rápida e resistência muscular com uma carga relativamente pequena. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: El método tradicional de entrenamiento de la fuerza muscular de las extremidades inferiores consiste principalmente en el entrenamiento de la resistencia, donde la intensidad del entrenamiento se incrementa gradualmente con el objetivo de ganar fuerza. Objetivo: Estudiar el efecto de diferentes frecuencias de vibración en el entrenamiento de la fuerza muscular de las articulaciones de la rodilla de los tenistas. Métodos: Utilizando la plataforma de entrenamiento de vibración PHYSIO-PLATE, los jugadores de tenis de un equipo de Pekín fueron sometidos a diferentes frecuencias de entrenamiento de fuerza con estimulación de vibración, después de ocho semanas de entrenamiento de fuerza sistemático, las frecuencias de vibración fueron 30Hz y 45Hz, con amplitud de 7mm. Resultados: Después del experimento, el par máximo relativo y el trabajo total de los músculos extensores de la rodilla en los sujetos de los grupos I y II mejoraron significativamente (P<0,05), generando un aumento significativo en el inicio de la potencia máxima rápida. Conclusión: La adición de la estimulación vibratoria al entrenamiento de la fuerza muscular puede mejorar eficazmente su efecto, incluyendo características como la fuerza máxima, la fuerza rápida y la resistencia muscular con una carga relativamente pequeña. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Integrated Bioinformatics Analysis Identifies Robust Biomarkers and Its Correlation With Immune Microenvironment in Nonalcoholic Fatty Liver Disease.

Objective: Nonalcoholic fatty liver disease (NAFLD) is a serious threat to human health worldwide. In this study, the aim is to analyze diagnosis biomarkers in NAFLD and its relationship with the immune microenvironment based on bioinformatics analysis. Methods: We downloaded microarray datasets (GSE48452 and GSE63067) from the Gene Expression Omnibus (GEO) database for screening differentially expressed genes (DEGs). The hub genes were screened by a series of machine learning analyses, such as support vector machine (SVM), least absolute shrinkage and selection operator (LASSO), and weighted gene co-expression network analysis (WGCNA). It is worth mentioning that we used the gene enrichment analysis to explore the driver pathways of NAFLD occurrence. Subsequently, the aforementioned genes were validated by external datasets (GSE66676). Moreover, the CIBERSORT algorithm was used to estimate the proportion of different types of immune cells. Finally, the Spearman analysis was used to verify the relationship between hub genes and immune cells. Results: Hub genes (CAMK1D, CENPV, and TRHDE) were identified. In addition, we found that the pathogenesis of NAFLD is mainly related to nutrient metabolism and the immune system. In correlation analysis, CENPV expression had a strong negative correlation with resting memory CD4 T cells, and TRHDE expression had a strong positive correlation with naive B cells. Conclusion: CAMK1D, CENPV, and TRHDE play regulatory roles in NAFLD. In particular, CENPV and TRHDE may regulate the immune microenvironment by mediating resting memory CD4 T cells and naive B cells, respectively, and thus influence disease progression.

Colorectal leiomyosarcoma with BRCA2 mutation benefit from treatment with olaparib: a case report.

Background: Colorectal leiomyosarcoma (LMS) is a rare colorectal malignancy accounting for approximately 1% of all colorectal malignancies with a poor prognosis and limited treatment options. Targeted therapies have been applied for breast cancer 2 (BRCA2) alterations, but their role remains to be explored in colorectal LMS. This case could provide clinical proof for the application of olaparib for LMS patients. Case Description: Here, we present a case of colorectal LMS with BRCA2 alterations who was treated with olaparib and achieved progression-free survival (PFS) for 1 year. In August 2016, a 46-year-old female patient was admitted to hospital due to a mass in the left lower abdomen and was diagnosed with LMS of the sigmoid colon. After surgical resection, chemotherapy with ifosfamide or ifosfamide combined with pirarubicin was given and achieved stable disease (SD) until the disease progressed 1.5 years later. Afterwards, a multi-target tyrosine kinase inhibitor, anlotinib, was taken. Before the observation of lung and liver metastasis, the patient's disease was stable for 1 year. BRCA2 mutation and rearrangement was revealed by next-generation sequencing (NGS), and the targeted therapy, olaparib, was given. Efficacy evaluation showed SD for 1 year, and no obvious toxic and side effects were observed. Conclusions: Our case suggested that NGS should be considered for further treatment of patients with colorectal LMS, and poly (ADP-ribose) polymerase (PARP) inhibitors could be a feasible therapy for LMS patients with BRCA2 alterations.

Genomic landscape and prognosis of patients with TP53-mutated non-small cell lung cancer.

Background: The TP53 tumor suppressor gene plays an important role in preventing and inhibiting the growth of tumor by regulating cell cycle, apoptosis and DNA repair. Meanwhile, the TP53 gene is one of the most frequently altered gene in non-small cell lung cancer (NSCLC) patients. Mutant TP53 (TP53-MUT) may lose tumor suppressor activity and gain tumor promoting functions, which play an important role in cancer risk, therapy resistance and poor prognosis. The impact of TP53-MUT on the prognosis of NSCLC patients need to be further studied. Methods: We obtained genomic and clinical data from The Cancer Genome Atlas (TCGA). Mutation profiles, the TMB, disease-free survival (DFS), and overall survival (OS) were compared between patients with different TP53-MUT statuses. Results: TP53-MUTs were detected in 46.6% of patients with lung adenocarcinoma (LUAD) (264 of 566) and 82.3% of those with lung squamous cell carcinoma (LUSC) (401 of 487). The most frequently co-mutated genes in patients with LUAD carrying a TP53-MUT included classic driver genes such as epidermal growth factor receptor (EGFR) and anaplastic large-cell lymphoma kinase (ALK), while Kirsten rat sarcoma viral oncogene (KRAS) mutations and TP53-MUTs appear to be mutually exclusive. This mutual exclusivity was not observed in patients with LUSC, in whom titin (TTN) and CUB and Sushi multiple domains 3 (CSMD3) were the most frequently co-mutated genes. A higher TMB was significantly associated with TP53-MUTs in patients with LUAD but not in those with LUSC. In patients with stage I-III NSCLC who had undergone surgery, there was no significant difference in DFS between patients carrying TP53-wildtype (TP53-WT) and TP53-MUTs, irrespective of histology or mutation type. However, the presence of TP53-MUT was associated with shorter OS in patients with LUAD (49 vs. 54 months, respectively; P=0.13) and significantly longer OS in those with LUSC (62 vs. 29 months, respectively; P=0.015). Conclusions: In contrast to most previous studies, we revealed TP53-MUT characteristic in NSCLC patients according to histology-specific differences and the association between TP53-MUT and the mutation landscape, the TMB, and the OS. These findings suggest a need for individualized management for patients with LUAD and LUSC who carry a TP53-MUT, and warrant further research.

Analysis of Body Behavior Characteristics after Sports Training Based on Convolution Neural Network.

The use of artificial intelligence technology to analyze human behavior is one of the key research topics in the world. In order to detect and analyze the characteristics of human body behavior after training, a detection model combined with a convolutional neural network (CNN) is proposed. Firstly, the human skeleton suggestion model is established to analyze the driving mode of the human body in motion. Secondly, the number of layers and neurons in CNN are set according to the skeleton feature map. Then, the output information is classified according to the fatigue degree according to the body state after exercise. Finally, the training and performance test of the model are carried out, and the effect of the body behavior feature detection model in use is analyzed. The results show that the CNN designed in the study shows high accuracy and low loss rate in training and testing and also has high accuracy in the practical application of fatigue degree recognition after human training. According to the subjective evaluation of volunteers, the overall average evaluation is more than 9 points. The above results show that the designed convolution neural network-based detection model of body behavior characteristics after training has good performance and is feasible and practical, which has guiding significance for the design of sports training and training schemes.

MicroRNA­485­5p suppresses the progression of esophageal squamous cell carcinoma by targeting flotillin­1 and inhibits the epithelial­mesenchymal transition.

As esophageal squamous cell carcinoma (ESCC) is one of the most frequently diagnosed cancers in Asia, it is crucial to uncover its underlying molecular mechanisms that support its development and progression. Several articles have reported that microRNA (miR)­485­5p inhibits the malignant phenotype in a number of cancer types, such as lung, gastric and breast cancer, but to the best of our knowledge, its function in ESCC has not been studied in depth until the present study. It is of great significance to probe the regulatory action and underlying mechanism of miR­485­5p in ESCC. In brief, this study identified that miR­485­5p expression in ESCC tissues was significantly lower than that in normal tissues. The decrease in miR­485­5p expression was associated with a larger tumour size and poor histology and stage. The expression of miR­485­5p was relatively high in Eca 109 and TE­1 cells, but relatively low in KYSE 30. The overexpression of miR­485­5p inhibited cell proliferation, migration and invasion in vitro, whereas miR­485­5p knockdown did the opposite. Flotillin­1 (FLOT­1) can facilitate the malignant phenotype in various cancer types. The present study found that in ESCC tissue, the protein expression of FLOT­1 was negatively correlated with miR­485­5p expression. Further experiments showed that miR­485­5p directly targeted the 3'­untranslated region of FLOT­1. The overexpression of miR­485­5p significantly suppressed the mRNA and protein expression levels of FLOT­1, whereas knockdown had the reverse effects. Furthermore, overexpression of miR­485­5p restrained epithelial­mesenchymal metastasis (EMT)­related factors at both the mRNA and protein levels. At the same time, it also inhibited the growth of ESCC and restrained the EMT in vivo. In summary, miR­485­5p was found to be an inhibitor of ESCC and may have potential as a novel target candidate for ESCC treatment.

The Prognostic Value of Baseline Distant Metastasis in Icotinib-Treated Patients with EGFR-Mutated Stage IV Non-Small Cell Lung Cancer.

PURPOSE: Several studies have revealed the prognostic value distant metastasis in non-small-cell lung cancer (NSCLC) patients receiving first-line epidermal growth factor receptor (EGFR) inhibitors. However, the question of whether the specific metastatic site could predict survival outcomes remain elusive. This study evaluated the prognostic value of specific metastatic site at diagnosis in first-line icotinib-treated patients with EGFR-mutated advanced NSCLC. METHODS: A total of 216 patients with EGFR-mutated stage IV NSCLC who received first-line icotinib treatment were retrospectively enrolled. The associations between the presence of distant metastasis to certain organs at diagnosis and survival outcomes were analyzed. PATIENTS AND METHODS: The presence of distant metastases was not associated with progression-free survival. Patients with liver metastasis showed a significantly shorter OS than those without liver metastasis (14.6m vs 33.0m, p=0.024). Patients with brain metastasis showed a marginally shorter OS than those without brain metastasis (26.5m vs 33.8m, p=0.051). Patients with lung metastasis showed a significantly longer OS than those without lung metastasis (36.0m vs 28.6m, p=0.038). Multivariable Cox regression analysis showed the presence of liver metastasis (HR [hazard ratio]: 2.265, 95% CI [confidence interval]: 1.239-4.139, p=0.008) and brain metastasis (HR: 1.493, 95% CI: 1.012-2.202, p=0.043) were independent predictors for unfavorable OS, while lung metastasis (HR: 0.669, 95% CI: 0.460-0.971, p=0.034) was an independent predictor for favorable OS. CONCLUSION: The presence of liver and brain metastasis predicted unfavorable OS, while the presence of lung metastasis predicted favorable OS in first-line icotinib-treated patients with EGFR-mutated stage IV NSCLC.

SNHG5 inhibits the progression of EMT through the ubiquitin-degradation of MTA2 in oesophageal cancer.

A substantial fraction of transcripts are known as long noncoding RNAs (lncRNAs), and these transcripts play pivotal roles in the development of cancer. However, little information has been published regarding the functions of lncRNAs in oesophageal squamous cell carcinoma (ESCC) and the underlying mechanisms. In our previous studies, we demonstrated that small nucleolar RNA host gene 5 (SNHG5), a known lncRNA, is dysregulated in gastric cancer (GC). In this study, we explored the expression and function of SNHG5 in development of ESCC. SNHG5 was found to be downregulated in human ESCC tissues and cell lines, and this downregulation was associated with cancer progression, clinical outcomes and survival rates of ESCC patients. Furthermore, we also found that overexpression of SNHG5 significantly inhibited the proliferation, migration and invasion of ESCC cells in vivo and in vitro. Notably, we found that metastasis-associated protein 2 (MTA2) was pulled down by SNHG5 in ESCC cells using RNA pulldown assay. We also found that SNHG5 reversed the epithelial-mesenchymal transition by interacting with MTA2. In addition, overexpression of SNHG5 downregulated the transcription of MTA2 and caused its ubiquitin-mediated degradation. Thus, overexpression of MTA2 partially abrogated the effect of SNHG5 in ESCC cell lines. Furthermore, we found that MTA2 mRNA expression was significantly elevated in ESCC specimens, and a negative correlation between SNHG5 and MTA2 expression was detected. Overall, this study demonstrated, for the first time, that SNHG5-regulated MTA2 functions as an important player in the progression of ESCC and provide a new potential therapeutic strategy for ESCC.

Effect of DUOX2 on sensitivity of colorectal cancer cells to 5-fluorouracil / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of double oxidase 2 (DUOX2) on the sensitivity of colorectal cancer (CRC) cells to 5-fluorouracil (5-FU). Methods: CRC cell lines DLD-1, SW480, HCT116, SW620 and normal intestinal epithelial cell line NCM460 were selected, and the expression of DUOX2 in these cell lines were detected by qPCR. DUOX2 expression in HT-29 and HCT116 cells was stably knocked down by lentivirus infection technique. The knockdown efficiency was detected by qPCR and WB. Cells in sh-Control and sh-DUOX2 groups were treated with 5-FU at different concentrations (0, 5, 10, 20, 40, 80, 120 μg/ml). The effects of 5-FU on cell proliferation, apoptosis and cell cycle were detected by CCK-8 method and flow cytometry. HT29 cell transplanted xenograft model in nude mice was constructed to observe the effect of DUOX2 gene on the treatment efficacy of 5-FU. Results: the expression level of DUOX2 mRNA in CRC cells was significantly higher than that in NCM460 cells (P<0.05 or P<0.01). Compared with sh-Control group, the mRNA and protein expressions of DUOX2 in sh-DUOX2 group were significantly decreased (all P<0.01); the sensitivity of cells to 5-FU was enhanced, the apoptosis rate and the ratio of cells at G0/G1 phase were significantly increased (all P<0.01), and the ratio of cells at G2 and S phase was significantly decreased (all P<0.01). There was no significant difference in tumor volume and mass between sh-Control group and sh-DUOX2 group without 5-FU treatment (all P>0.05), but the volume and mass of transplanted tumor in sh-DUOX2+5-FU group after 5-FU treatment was significantly lower than that in sh-Control+5-FU group (all P<0.01). Conclusion: The sensitivity of CRC cells to 5-FU can be significantly enhanced by knocking down DUOX2 gene.

V3S4 Nanosheets Anchored on N, S Co-Doped Graphene with Pseudocapacitive Effect for Fast and Durable Lithium Storage.

Construction of a suitable hybrid structure has been considered an important approach to address the defects of metal sulfide anode materials. V3S4 nanosheets anchored on an N, S co-coped graphene (VS/NSG) aerogel were successfully fabricated by an efficient self-assembled strategy. During the heat treatment process, decomposition, sulfuration and N, S co-doping occurred. This hybrid structure was not only endowed with an enhanced capability to buffer the volume expansion, but also improved electron conductivity as a result of the conductive network that had been constructed. The dominating pseudocapacitive contribution (57.78% at 1 mV s-1) enhanced the electrochemical performance effectively. When serving as anode material for lithium ion batteries, VS/NSG exhibits excellent lithium storage properties, including high rate capacity (480 and 330 mAh g-1 at 5 and 10 A g-1, respectively) and stable cyclic performance (692 mAh g-1 after 400 cycles at 2 A g-1).

CIK cell-based delivery of recombinant adenovirus KGHV500 carrying the anti-p21Ras scFv gene enhances the anti-tumor effect and safety in lung cancer.

PURPOSE: Adenovirus (Ads) is one of the most popular vectors used in gene therapy for the treatment of cancer. However, systemic therapy is limited by circulating antiviral antibodies and poor viral delivery in vivo. In this study, we used cytokine-induced killer (CIK) cells as delivery vehicles of Ads KGHV500 carrying the anti-p21Ras scFv gene to treat Ras gene-related lung cancer and investigate the anti-tumor effect in vitro and in vivo. METHODS: The human lung cancer cell line A549 was employed to investigate the anti-tumor activity of recombinant Ads KGHV500 harboring the anti-p21Ras scFv gene using MTT, wound healing, transwell invasion, and apoptosis assays in vitro. Next, CIK cells were used as delivery vehicles to deliver KGHV500 carrying the anti-p21Ras scFv gene to treat A549-transplanted tumors in nude mice, and viral replication, p21Ras scFv expression, and the therapeutic efficacy were assessed. RESULTS: In vitro studies showed that KGHV500 had potent anti-tumor activity. In addition, in vivo, this combination therapy significantly inhibited the growth of lung cancer xenografts compared with mice treated with KGHV500 alone. KGHV500 and anti-p21Ras scFv were observed in tumor tissue, but were nearly undetectable in normal tissues. CONCLUSIONS: The co-delivery of anti-p21Ras scFv by CIK cells and KGHV500 could increase the anti-tumor effect and safety, and possess considerable advantages for the treatment of Ras-related cancer.

miR-200c inhibits TGF-ß-induced-EMT to restore trastuzumab sensitivity by targeting ZEB1 and ZEB2 in gastric cancer.

Gastric cancer is the fifth most common malignancy in the world, with Eastern Asia as one of areas with the highest incidence rates. Trastuzumab, a HER2-targeting antibody, combined with chemotherapy has been successfully employed for the gastric cancer patients with HER2 overexpression/amplification. However, trastuzumab resistance is a major problem in clinical practice. Here we observed that the trastuzumab-resistant gastric cancer cell line NCI-N87/TR expressed high levels of epithelial-mesenchymal transition factors and demonstrated increased migration and invasion capability compared with NCI-N87 cells. Downregulated E-cadherin and increased N-cadherin, TGF-ß, ZEB1, ZEB2, TWIST1, and Snail were detected in NCI-N87/TR cells. We also found that miR-200c was downregulated in NCI-N87/TR cells compared with parental cells NCI-87 by qRT-PCR. Treatment with TGF-ß downregulated the expression of miR-200c and upregulated ZEB2, and significantly decreased the trastuzumab sensitivity of NCI-N87 cells. miR-200c restored trastuzumab sensitivity and inhibited migration and invasion through suppressing ZEB1 and ZEB2. In summary, TGF-ß/ZEB2 axis plays an encouraging role in trastuzumab resistance of gastric cancer, while miR-200c overexpression downregulates ZEB1/ZEB2 and resensitizes drugs resistance. Our findings might provide a potential therapeutic strategy for trastuzumab resistance of gastric cancer.

Flotillin1 promotes EMT of human small cell lung cancer via TGF-ß signaling pathway.

OBJECTIVE: Small cell lung carcinoma (SCLC) is considered one of the most aggressive types of lung cancer due to its rapid growth and early metastasis. No tumor markers or therapeutic targets have been demonstrated to be specific or effective in SCLC to date. This study aims to evaluate the potential of Flotillin1 (Flot1) as a target of SCLC treatment. METHODS: Flot1 expression level in the tissue of SCLC and other tissue of lung disease was detected using immunohistochemical staining. Transwell and Matrigel assays were employed to examine migration and invasion of cancer cells. Flow cytometry and xCELLigence system were used to evaluate cell apoptosis and cell viability, respectively. Expression levels of Flot1, epithelial-mesenchymal transition (EMT) marker E-cadherin, vimentin, cyclinD1, TGF-ß-Smad2/3, and p-AKT were examined using Western blot. Furthermore, xenograft tumor in nude mice was used to evaluate the growth and metastasis of NCI-H446 cells in vivo. RESULTS: Our results demonstrated that Flot1 is highly expressed in SCLC samples and that its expression correlates strongly with clinical stage, distant metastasis, and poor survival. The knockdown of Flot1 decreased the growth, migration, and invasiveness of SCLC cells and reversed EMT phenotype in vitro and in vivo, while enhanced Flot1 expression exhibited the opposite behavior. Gene expression profile analysis demonstrated that Flot1-regulated genes frequently mapped to the AKT and TGF-ß-Smad2/3 pathways. Our results further revealed that Flot1 affected the progression of SCLC via regulation of EMT progression. CONCLUSIONS: These findings indicated an oncogenic role of Flot1 via promoting EMT in SCLC and suggested its potential as a tumor marker and prognostic indicator.

Expression of miR-133a-3p in gastric cancer tissues and plasma and its effect on proliferation of gastric cancercells / 中国肿瘤生物治疗杂志

@#Objective: To detect the expression of miR-133a-3p in gastric cancer (GC) tissues and plasma of GC patients, and to investigate its effect on the proliferation of GC cells as well as its correlation toprognosis of GC patients. Methods: 52 cases of cancertissues (non-necrosis part) and corresponding adjacent tissues as well as the pre-operative peripheral blood samples from GC patients, who underwent surgery at Department of General Surgery, the Forth Hospital of Hebei Medical University(Shijiazhuang, China) between May 2012 and May 2013, were collected for this study. The plasma sample (n=35) from healthy donors were obtained during their physical examination. RT-qPCR was adopted to detect the expression of miR-133a-3p in gastric cancer tissues, adjacent tissuesand plasma samples of GC patients and healthy volunteers. The relationships between miR-133a-3p expression and the median DFS as well as clinicopathological parameters were also analyzed. CCK-8 assay was adopted to detect the effect of miR-133a-3p silence or over-expression on proliferation of gastric cancer SGC7901 cells. Results: miR-133a-3p was dramatically decreased in gastric cancer tissues (P<0.01), and its expression was associated with TNM stage, tumor infiltration (T), lynphonode metastasis (N), and vascular tumor thrombus (all P<0.01); miR-133a-3p was significantly increased in the plasma of GC patients (P<0.01), and its expression was associated with TNM stage, lynphonode metastasis (N), and vascular tumor thrombus (all P<0.05). miR-133a-3p expression was positively correlated with serum CA199 level of GC patients (P<0.01). The median DFS of patients with high miR-133a-3pexpression in cancer tissues was significantly longer than that of the patients with low expression(20.8 vs 14.8 months, P<0.05); The median DFS of patients with high plasma miR-133a-3p expression was significantly shorter than that of the patients with low expression (14.4 vs 20.3 months, P<0.05). Over-expression of miR-133a-3p could significantly inhibit the proliferation of gastric cancer SGC7901 cells, while miR-133a-3p silence could significantly promote the proliferation (all P<0.05). Conclusion: miR-133a-3p could significantlyinhibit the proliferation of SGC7901 cells; miR-133a-3p aberrantlyexpressed in gastric cancer tissues and plasma, and obviously correlated with prognosis of gastric cancer patients, which may be used as a potential clinical bio-maker for early diagnosis and treatment as well as the prognosis prediction of gastric cancer.

Overexpression of wild-type p21Ras plays a prominent role in colorectal cancer.

Colorectal cancer (CRC) is the most common gastrointestinal type of cancer. The overexpression of Ras proteins, particularly p21Ras, are involved in the development of CRC. However, the subtypes of the p21Ras proteins that are overexpressed and the mutation status remain unknown restricting the development of therapeutic antibodies targeting p21Ras proteins. The present study aimed to investigate the mutation status of ras genes associated with Ras proteins that are overexpressed in CRC and explore whether or not wild-type p21Ras could be a target for CRC therapy. p21Ras expression was examined immunohistochemically in normal colorectal epithelium, benign lesions and malignant colorectal tumor tissues by monoclonal antibody (Mab) KGH-R1 which is able to react with three types of p21Ras proteins: H-p21Ras, N-p21Ras and K-p21Ras. Then, the expression levels of p21Ras subtypes were determined in CRC by a specific Mab for each p21Ras subtype. Mutation status of ras genes in p21Ras-overexpressing CRC was detected by DNA sequencing. There was rare p21Ras expression in normal colorectal epithelium but a high level of p21Ras expression in CRC, with a significant increase from normal colorectal epithelium to inflammatory polyps, low-grade intraepithelial neoplasia, high-grade intraepithelial neoplasia and invasive colorectal adenocarcinoma, respectively. Overexpression of K-p21Ras was found in all CRC tissues tested, overexpression of N-p21Ras was found in 85.7% of the CRC tissues, while H-p21Ras expression was not found in any CRC tissue. DNA sequencing showed that there were no K-ras mutations in 60% of the K-p21Ras-overexpressing CRC, while 40% of the CRC tissues harbored K-ras mutations. N-ras mutations were not found in any N-p21Ras-overexpressing CRC. Our findings indicate that overexpression of wild-type p21Ras may play a prominent role in the development of CRC in addition to ras mutations and could be a promising target for CRC therapy.

Effects of Combined Simultaneous and Sequential Endostar and Cisplatin Treatment in a Mice Model of Gastric Cancer Peritoneal Metastases.

Objective. Aimed to study the effects of endostar and cisplatin using an in vivo imaging system (IVIS) in a model of peritoneal metastasis of gastric cancer. Methods. NUGC-4 gastric cancer cells transfected with luciferase gene (NUGC-4-Luc) were injected i.p. into nude mice. One week later, mice were randomly injected i.p.: group 1, cisplatin (d1-3) + endostar (d4-7); group 2, endostar (d1-4) + cisplatin (d5-7); group 3, endostar + cisplatin d1, 4, and 7; group 4, saline for two weeks. One week after the final administration, mice were sacrificed. Bioluminescent data, microvessel density (MVD), and lymphatic vessel density (LVD) were analyzed. Results. Among the four groups, there were no significant differences in the weights and in the number of cancer cell photons on days 1 and 8 (P > 0.05). On day 15, the numbers in groups 3 and 1 were less than that in group 2 (P < 0.05). On day 21, group 3 was significantly less than group 2 (P < 0.05). MVD of group 4 was less than that of groups 1 and 2 (P < 0.01). There was no significant difference between groups 2 and 3 (P > 0.05) or in LVD number among the four groups (P > 0.05). Conclusions. IVIS® was more useful than weight, volume of ascites, and number of peritoneal nodules. The simultaneous group was superior to sequential groups in killing cancer cells and inhibiting vascular endothelium. Cisplatin-endostar was superior to endostar-cisplatin in killing cancer cells, while the latter in inhibiting peritoneal vascular endothelium.

Role of microRNA-141 in colorectal cancer with lymph node metastasis.

The present study aimed to investigate the role of microRNA (miR)-141 in the pathogenesis of colorectal cancer (CRC). In total, 58 CRC patients were included in the present study. The mRNA and protein expression levels of mitogen-activated protein kinase 4 (MAP4K4) were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis, respectively. The miRNA-141 expression was measured by RT-qPCR, while serum MAP4K4 content was detected by enzyme-linked immunosorbent assay. Natural killer (NK) cells and T cells in peripheral blood were detected by flow cytometry. The results indicated that the mRNA and protein expression levels of MAP4K4 were significantly elevated in the tumor tissues, lymph nodes (P<0.01) and serum (P<0.05) in CRC. Furthermore, the expression levels of MAP4K4 in CRC patients with lymph node metastasis were higher compared with those in patients without metastasis. Bioinformatics analysis revealed that MAP4K4 may be the target gene of miRNA-141. The expression levels of miRNA-141 in the tumor tissues, lymph nodes and serum were significantly decreased in CRC patients, with a more evident decline in cases with lymph node metastasis. In addition, the percentage of NK, CD3+ T and CD4+ T cells was significantly decreased, whilst the number of CD8+ T cells was significantly increased, in the peripheral blood in CRC. The present results showed that miRNA-141 was downregulated in CRC, which increased the expression levels of MAP4K4 and altered the anti-tumor response, further increasing the proliferation, invasion and metastasis of the tumors. These findings may contribute to improving the current understanding of the pathogenesis of CRC, and lead to the development of therapies involving miRNA-141.