bluuebunny/crossref_metadata_2025_split huggingface dataset:
DNMT1 facilitates proliferation and metastasis of breast cancer by inducing MEG3 promoter methylation in MEG3/miR-494-3p/OTUD4 regulatory axis
Authors: Zhu Xiaotao , Lv Lin , Wang Mingzheng , Fan Chen , Lu Xiaofeng , Jin Miaomiao , Li Shuguang , Wang Fan 
Published: December 2020

Abstract
Background: To explore the mechanism by which DNMT1 potentiates proliferation and metastasis of breast cancer by inducing MEG3 promoter methylation in MEG3/miR-494-3p/OTUD4 regulatory axis.Methods: Human breast cancer cell lines (MCF-7, MDA-MB-231, SKBR3) and human breast epithelial cell line MCF10A were selected for the experiments. The expression levels of DNMT1, MEG3, miR-494-3p and OTUD4 were detected by qRT-PCR. Western blot was used to detect the protein expression levels of DNMT1 and OTUD4. ChIP assay was adopted to verify the binding relationship between DNMT1 and MEG3 promoter region. MethPrimer software was applied to identify MEG3 promoter methylation while methylation-specific PCR was conducted to examine its methylation level. The targeted binding sites of miR-494-3p on MEG3/OTUD4 were predicted by bioinformatics and further verified by RNA binding protein immunoprecipitation assay plus dual-luciferase reporter gene assay. Cell proliferative, migratory and invasive abilities were measured by CCK-8, wound healing and Transwell assays.Results: DNMT1 was highly expressed while MEG3 was poorly expressed in breast cancer cells. Silencing DNMT1 inhibited the proliferation, migration and invasion of breast cancer cells by increasing gene expression of MEG3 through demethylation. MEG3 was identified as a ceRNA that regulated miR-494-3p expression via RNA sponging in breast cancer. In addition, miR-494-3p could bind to the 3’-UTR of OTUD4 mRNA, thus negatively regulating OTUD4 expression. A regulator axis formed by MEG3/miR-494-3p/OTUD4 was thus established and identified to have an impact on proliferation, invasion and migration of breast cancer. Moreover, overexpression of MEG3 could suppress tumor growth of breast cancer in vivo.Conclusion: Silencing DNMT1 induced demethylation of MEG3 promoter to promote MEG3 gene expression, in turn inhibiting the expression of miR-494-3p to elevate the expression of its downstream target OTUD4, thus weakening the proliferative, migratory and invasive abilities of breast cancer cells.
SKP1 promotes YAP-mediated colorectal cancer stemness via suppressing RASSF1
Authors: Tian Cong , Lang Tingyuan , Qiu Jiangfeng , Han Kun , Zhou Lei , Min Daliu , Zhang Zhiqi , Qi Dachuan 
Published: December 2020

Abstract
Background: Cancer stem cells (CSCs) have been recognized as an important drug target, however, the underlying mechanisms have not been fully understood. SKP1 is a traditional drug target for cancer therapy, while, whether SKP1 promotes colorectal cancer (CRC) stem cells (CRC-SCs) and the underlying mechanisms have remained elusive. Methods: Human CRC cell lines and primary human CRC cells were used in this study. Gene manipulation was performed by lentivirus system. The mRNA and protein levels of target genes were examined by qRT-PCR and western blot. The sphere-forming and in vitro migration capacities were determined by sphere formation and transwell assay. The self-renewal was determined by limiting dilution assay. The tumorigenicity and metastasis of cancer cells were examined by xenograft model. The promoter activity was examined by luciferase reporter assay. Nuclear run-on and Chromatin immunoprecipitation-PCR (ChIP-PCR) assay were employed to examine the transcription and protein-DNA interaction. Co-immunoprecipitation assay was used to test protein-protein interaction. The relationship between gene expression and survival was analyzed by Kaplan-meier analysis. The correlation between two genes was analyzed by Spearman analysis. Data are represented as mean ± s.d. and the significance was determined by Student’s t-test.Results: SKP1 was upregulated in CRC-SCs and predicted poor prognosis of colon cancer patients. Overexpression of SKP1 promoted the stemness of CRC cells reflected by increased sphere-forming, migration and self-renewal capacities as well as the expression of CSCs markers. In contrast, SKP1 depletion produced the opposite effects. SKP1 strengthened YAP activity and knockdown of YAP abolished the effect of SKP1 on the stemness of CRC cells. SKP1 suppressed RASSF1 at both mRNA and protein level. Overexpression of RASSF1 abolished the effect of SKP1 on YAP activity and CRC stemness. Conclusion: Our results demonstrated that SKP1 suppresses RASSF1 at both mRNA and protein level, attenuates Hippo signaling, activates YAP, and thereby promoting the stemness of CRC cells.
Assessment of mitochondrial genomes for heterobranch gastropod phylogenetics
Authors: Varney Rebecca M , Brenzinger Bastian , Malaquias Manuel António E. , Meyer Christopher P. , Schrödl Michael , Kocot Kevin 
Published: December 2020

Abstract
Background Heterobranchia is a diverse clade of marine, freshwater, and terrestrial gastropod molluscs. It includes such disparate taxa as nudibranchs, sea hares, bubble snails, pulmonate land snails and slugs, and a number of (mostly small-bodied) poorly known snails and slugs collectively referred to as the “lower heterobranchs.” Evolutionary relationships within Heterobranchia have been challenging to resolve and the group has been subject to frequent and significant taxonomic revision. Mitochondrial (mt) genomes can be a useful molecular marker for phylogenetics but, to date, sequences have been available for only a relatively small subset of Heterobranchia. Results To assess the utility of mitochondrial genomes for resolving evolutionary relationships within this clade, eleven new mt genomes were sequenced including representatives of several groups of “lower heterobranchs.” Maximum likelihood analyses of concatenated matrices of the thirteen protein coding genes found weak support for most higher-level relationships even after several taxa with extremely high rates of evolution were excluded. Bayesian inference with the CAT+GTR model resulted in a reconstruction that is much more consistent with the current understanding of heterobranch phylogeny. Notably, this analysis recovered Valvatoidea and Orbitestelloidea in a polytomy with a clade including all other heterobranchs, highlighting these taxa as important to understanding early heterobranch evolution. Also, dramatic gene rearrangements were detected within and between multiple clades. However, a single gene order is conserved across the majority of heterobranch clades.Conclusions Analysis of mitochondrial genomes in a Bayesian framework with the site heterogeneous CAT+GTR model resulted in a topology largely consistent with the current understanding of heterobranch phylogeny. However, mitochondrial genomes appear to be too variable to serve as good phylogenetic markers for robustly resolving a number of deeper splits within this clade.
Clinical features of Japanese patients with exacerbations of chronic obstructive pulmonary disease
Authors: Tashiro Hiroki , Kurihara Yuki , Takahashi Koichiro , Sadamatsu Hironori , Haraguchi Tetsuro , Tajiri Ryo , Takamori Ayako , Kimura Shinya , Sueoka-Aragane Naoko 
Published: December 2020

Abstract
BackgroundExacerbations are critical events in chronic pulmonary obstructive disease (COPD). The frequency of COPD exacerbations is associated with the prognosis, including mortality, but no useful biomarker has been established.MethodsThe present retrospective study investigated 481 COPD patients. Clinical features in the stable period were compared between patients who experienced severe exacerbation (n=88, 18.3%) and those who never experienced severe exacerbation (n=393, 81.7%). In the patients who experienced exacerbations, clinical features were also compared between frequent exacerbators (exacerbation rate ≥ 2 times/year, n=27, 30.7%) and infrequent exacerbators (1 time/year, n=61, 69.3%).ResultsCompared to COPD patients who never experienced exacerbations, body mass index (BMI), serum albumin, and pulmonary functions were significantly lower, and the cardiovascular disease comorbidity rate, COPD assessment test score, modified Medical Research Council dyspnea scale, and use of long-term oxygen therapy, long-acting β2 adrenergic agonist therapy, inhaled corticosteroid therapy, and macrolide therapy were significantly higher in COPD patients with exacerbations (all p < 0.01). In patients who experienced exacerbations, frequent exacerbators had significantly lower % forced expiratory volume in 1.0 second and a higher risk of critical exacerbations, percentage of blood eosinophils, history of mechanical ventilation use, and use of long-term oxygen therapy and of macrolide therapy than infrequent exacerbators (all p < 0.01). On multivariate analysis, the percentage of blood eosinophils was the parameter most correlated with exacerbation frequency (β value [95% confidence interval] 1.45 [1.12 – 1.88], p < 0.01).ConclusionBlood eosinophil in the stable period is the factor most correlated with the frequency of severe exacerbations.
Management of unexplained: poly-consultation and hyperfrequency on Primary Health Chilean Attention. A qualitative and exploratory study.
Authors: Sánchez Nicolás Fuster , López Diego Rivera , Retamales Hugo Sir , Pérez Constanza Gómez , Torres Magdalena Rodríguez 
Published: December 2020

Abstract
Background: In Europe, Latin-America, and Asia, poly-consultation has become a complex problem for the management of different healthcare systems. However, in the current literature, little attention has been paid to exploring perspectives of territorial and critical analysis to manage unexplained symptoms.The purpose of this study is to analyze the socio-structural elements that underlie the users’ phenomenon of poly-consultation or hyperfrequency in the Chilean primary healthcare system (PHCS). Methods: This paper represents qualitative data collected as part of an exploratory study that used mixed methods across three metropolitan areas of Santiago, Valparaíso, and Concepción, Chile. The study involved a sample of 24 subjects from administrative and management positions in PHC who were recruited from Family Health Care Centers, considering urban municipalities from the low, medium, and high stratum.The study collected data using one set of semi-standardized interviews during a year. Data analysis used an qualitative content analysis. Results: This article shows that poly-consultant patients provide a critical clinic category to management and mental health model (foreshadowing a social topology base that promotes a determinate unspecific and diffuse experience on users) that cannot be cover by current biomedical models. Data showed the strain of a somatoform clinic category, especially in the clinic and epistemological exercise, and, also, the relevance of particularities of Chile, a country with a mixed health system characteristics and their effects: the naturalization of collective problems managed as individual problems. Conclusions: Results from the study have the potential to inform healthcare professionals and managers of strategies for developing effective and territorially based. We conclude that hyperfrequency and poly-consultation in Chile reveal relevant stratification in the territory who has particularities than could be studied from a quantitative perspective.
Pilot Randomised Evaluation of Singing in Dementia (PRESIDE): protocol for a two-arm, parallel group randomised controlled feasibility study with waiting-list control
Authors: Dowson Becky , Schneider Justine , Guo Boliang , Bath Philip M , McDermott Orii , Haywood Lee J , Orrell Martin 
Published: December 2020

Abstract
BackgroundAs the number of people living with dementia grows, so does the need to provide them with adequate psychosocial support. Many people with dementia live at home with family carers, who also require social and emotional support to cope with their role. Community group singing has received attention for its potential to support people with dementia and their carers. It is postulated that singing can improve cognitive function, strengthen the bonds between care partners, and help to establish social support networks. However, there is a lack of rigorous evidence of singing’s benefits for this population. This study aims to test the feasibility of a randomised controlled trial of community singing in dementia, to pave the way for a larger, conclusive study.MethodsThe PRESIDE study is designed as a two-arm, parallel group randomised trial with a waiting list control. Dyads consisting of a person with dementia (n=80) and their carer (n=80) will be recruited. Each dyad will be randomised either to attend 10 weeks of community group singing sessions straight away, or to wait for three months before attending the sessions. The singing sessions will be led by experienced professional musicians and will last about 90 minutes, including time for socialising. The primary outcome of this study is the attainment of feasibility criteria around recruitment, retention and the acceptability of the waiting list control. Secondary outcomes include the quality of life, mood, cognition, and musical engagement of the person with dementia, and quality of life, mood, and experiences/challenges of the carer. These data will be collected during home visits at baseline, and three and six months post-baseline.DiscussionDespite growing public interest in the positive effects of singing, and encouraging findings from qualitative and non-randomised quantitative studies, there is a lack of rigorous evidence. This is the first randomised controlled trial of community group singing for people with dementia in Europe, to our knowledge. If the results favour a full trial, conclusively demonstrating the effectiveness of group singing could positively affect the opportunities available to community-dwelling people with dementia and their carers.Trial RegistrationUnique identification number in ISRCTN registry: ISRCTN10201482https://www.isrctn.com/ISRCTN10201482. Date registered: 12/05/2020
High-intensity focused ultrasound enhances the cisplatin sensitivity to human non-small cell lung cancer&nbsp;in vitro and&nbsp;in vivo
Authors: Wang Liwei , Fan Huijun , Wang Haiyan , Bai Yunlu , Zhou Wenyan 
Published: December 2020

Abstract
Background: The incidence and mortality of non-small cell lung cancer (NSCLC) rank first among malignant tumors worldwide. Cisplatin (CDDP) is currently the first-line chemotherapy drug used in clinical practice. However, the underlying mechanism that NSCLC cells are resistant to CDDP has not been fully elucidated. Therefore, it is urgent to explore the exact function of chemotherapy resistance and improve the anti-tumor effect of CDDP treatment. Method: We investigated the surviving fraction of H1299 and A549 cells treated with different High-intensity focused ultrasound (HIFU) and various CDDP concentrations. HIFU (400W/cm 2 ) and CDDP (10 μM) were selected to probe the further function in H1299 and A549 cells. Flow cytometry, MTT, and colony formation assays were performed to evaluate the apoptosis and proliferative effect of HIFU and CDDP on NSCLC cells. Transwell assays were used to analyze the migration and invasion abilities of NSCLC cells with HIFU and CDDP treatments. Platinum (Pt) accumulation was further measured in H1299 and A549 cells with different treatments. Finally, An NSCLC patient-derived xenograft model was used to explore the effect of HIFU and CDDP on NSCLC tumor growth. Results: HIFU combined with CDDP can markedly reduce the surviving rate and promote the apoptosis of NSCLC cells. Furthermore, co-treatment with HIFU and CDDP significantly inhibited the proliferation, colony formation, migration, and invasion of NSCLC cells compared to that with any single treatment. Moreover, the combined therapy can effectively promote Pt accumulation in NSCLC cells. Further functional analysis suggested that HIFU combined with CDDP can inhibit tumor growth in an NSCLC patient-derived xenograft model . Finally, the CDDP method effectively upregulated the expression level of apoptosis-related protein, cleaved-PARP, which could be further enhanced by the HIFU treatment. Conclusion: Our results revealed that HIFU enhances the anti-NSCLC effect of CDDP both in vitro and in vivo , providing a promising combination therapy for clinical NSCLC treatment.
Predominantly symplastic phloem unloading of photosynthates maintains efficient starch accumulation in the cassava storage roots (Manihot esculenta Crantz)
Authors: Pan Kun , Lu Cheng , Nie Peixian , Hu Meizhen , Zhou Xincheng , Chen Xin , Wang Wenquan 
Published: December 2020

Abstract
Background: It is well known that cassava (Manihot esculenta Crantz) highly efficiently accumulates starch in its storage roots, but how photosynthates are transported from the leaves to the phloem (especially how they are unloaded into parenchymal cells of storage roots) remains unclear. Methods: Here, we investigated the phloem unloading pattern of sucrose and its impact on the development of cassava storage roots through microstructural and physiological analyses and carboxyfluorescein (CF) and isotope C14 tracing. Expression profiling of genes involved in symplastic and apoplastic transport was performed, and their correlations with storage root yield were determined in populations according to enzyme activity, Western blotting analysis and transcriptome sequencing. Results: Carbohydrates are transported mainly in the form of sucrose, with more than 54.6% present in the stem phloem at any time. Sucrose was predominantly unloaded symplastically from the phloem into storage roots, but there was a shift from apoplastic to symplastic unloading accompanied by the onset of root swelling. Statistical data concerning the microstructure revealed an enrichment of plasmodesmata within sieve, companion and parenchyma cells in the developing storage roots of a cultivar but not in a wild ancestor. Tracing tests with CF verified the existence of a coplastid channel, and [14C]Suc demonstrated that the marked sucrose could rapidly diffuse into root parenchyma cells from phloem cells. The relatively high expression of genes encoding SuSy and associated proteins in the storage roots at the middle and late stages but not in the early stage of storage roots, primary fibrous roots or secondary fibrous roots and the inverse expression pattern of SUTs, CWI and SAI in these corresponding organs supported the presence of a symplastic sucrose unloading pathway. The transcriptome pattern of genes involved in symplastic unloading and their significantly positive correction with starch yield at the population level confirmed that symplastic sucrose transport is vitally important in the development of cassava storage roots. Conclusions: In this study, we revealed a predominantly symplastic phloem unloading pattern of sucrose in cassava storage roots. This pattern is essential for the efficient starch accumulation for sucrose transport between high-yielding varieties and low-yielding wild ancestors.
A Non-Synonymous Variant rs12614 of Complement Factor B Associated with Risk of Chronic Hepatitis B in a Korean Population
Authors: Seo Jung Yeon , Shin Joong Gon , Youn Byeong Ju , Namgoong Suhg , Cheong Hyun Sub , Kim Lyoung Hyo , Kim Ji On , Shin Hyoung Doo , Kim Yoon Jun 
Published: December 2020

Abstract
Background: Hepatitis B is known to cause several forms of liver diseases including chronic hepatitis B (CHB), and hepatocellular carcinoma. Previous genome-wide association study of CHB risk has demonstrated that rs12614 of complement factor B (CFB) was significantly associated with CHB risk. In this study, fine-mapping study of previously reported GWAS single nucleotide polymorphism (SNP; CFB rs12614) was performed to validate genetic effect of rs12614 on CHB susceptibility and identify possible additional causal variants around rs12614 in a Korean population. This association study was conducted in order to identify genetic effects of CFB single nucleotide polymorphisms (SNPs) and to identify additional independent CHB susceptible causal markers within a Korean population.Methods: A total of 10 CFB genetic polymorphisms were selected and genotyped in 1,716 study subjects comprised of 955 CHB patients and 761 population controls. Results: A non-synonymous variant, rs12614 (Arg32Trp) in exon2 of CFB, had significant associations with risk of CHB (odds ratio = 0.43, P = 5.91×10-10). Additional linkage disequilibrium and conditional analysis confirmed that rs12614 had independent genetic effect on CHB susceptibility with previously identified CHB markers. The genetic risk scores (GRSs) were calculated and the CHB patients had higher GRSs than the population controls. Moreover, OR was found to increase significantly with cumulative GRS. Conclusions: rs12614 showed significant genetic effect on CHB risk within the Korean population. As such rs12614 may be used as a possible causal genetic variant for CHB susceptibility.
Global transcriptome changes of elongating internode of sugarcane in response to mepiquat chloride
Authors: Chen Rongfa , Fan Yegeng , Zhou Huiwen , Mo Shanping , Zhou Zhongfeng , Yan Haifeng , Luo Ting , Huang Xing , Weng Mengling , Lakshmanan Prakash , Li Yangrui , Qiu Lihang , Wu Jianming 
Published: December 2020

Abstract
Background: Mepiquat chloride (DPC) is an extensively used chemical to control internode growth and compact canopies in cultured plants. Previous studies suggested that DPC could inhibit gibberellin biosynthesis in sugarcane. Unfortunately, the molecular mechanism underlying the suppressive effects of DPC on plant growth is still largely unknown. Results: In the present study, we first obtained the high-quality long transcripts from internode of sugarcane by PacBio Sequel System. A total of 72,671 isoforms with N50 as 3,073 were generated. These long isoforms were used for the following RNA-seq as reference. Subsequently, short reads generated from Illumina Hiseq 4000 platform were used for comparing the differentially expressed genes in DPC and control groups. The transcriptome profiling showed the 6 days post DPC treatment had the most significant changed genes. These genes were related to plant hormone signal transduction and biosynthesis of several metabolites, indicating DPC affected multiple pathways beside depressed gibberellin biosynthesis. The network of DPC on the key stage was illustrated by weighted gene co-expression network analysis (WGCNA). Among the constructed 36 modules, the top positive correlated module with stage of 6 day post spraying DPC was sienna3. Stf0 sulphotransferase, cyclin-like F-box and HOX12 were the hub genes in sienna3 that had high correlation with other genes in this module. The qPCR demonstrated the high accuracy of RNA-seq result. Conclusion: Taken together, we demonstrated the key role of these genes in DPC-induced growth inhibition in sugarcane.