MiR-183-5p promotes the progression of non-small cell lung cancer through targeted regulation of FOXO1

Objective To investigate miR-183-5p targeting to forkhead box protein O1(FOXO1)and its corresponding effect on the proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)of non-small cell lung cancer(NSCLC)cells.Methods NSCLC tissues and adjacent normal tissues from 60 patients with NSCLC adenocarcinoma were obtained via pathological biopsy or intraoperative resection.Several cell lines were cultured in vitro,including the human normal lung epithelial cell line BEAS-2B and human NSCLC cell lines A549,SPCA-1,PC-9,and 95-D.miR-183-5p and FOXO1 mRNA expression in tissues and cells were detected by qRT-PCR;the corresponding correlations in NSCLC tissues were analyzed using the Pearson test,and the relationship between miR-183-5p expression and clinicopathological parameters was analyzed.The miR-183-5p-mediated regulation of FOXO1 was verified by bioinformatics prediction alongside double luciferase,RNA-binding protein immunoprecipitation(RIP)assay,and pull-down experiments.A549 cells were divided into control,anti-miR-NC,anti-miR-183-5p,miR-NC,miR-183-5p,miR-183-5p+pcDNA3.1,and miR-183-5p+pcDNA3.1-FOXO1 groups.Cell proliferation,invasion,migration,apoptosis,and cell cycle distribution were detected using an MTT assay,clone formation assay,Transwell assay,scratch test,and flow cytometry,respectively.The expression of EMT-related proteins in the cells was analyzed by western blotting.The effect of miR-185-3p silencing on the development of transplanted tumors was detected by analyzing tumor formation in nude mice.Results miR-183-5p expression was significantly higher in NSCLC tissues and cells than in adjacent normal tissues,whereas FOXO1 mRNA expression was significantly down-regulated.There was a significant negative correlation between miR-183-5p and FOXO1 mRNA in NSCLC tissues(P<0.05).Additionally,the expression of miR-183-5p was significantly correlated with tumor size,tumor differentiation,and tumor-node-metastasis stage in patients with NSCLC(P<0.05).miR-183-5p targeted and inhibited FOXO1 expression.Compared to the anti-miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the anti-miR-183-5p group,whereas the protein expression of E-cadherin andα-catenin and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion were significantly lower in the anti-miR-183-5p group(P<0.05).Compared to the miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells in the miR-183-5p group were significantly higher,whereas the E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly lower;furthermore,the frequency of colony formation and invasion were significantly higher in the miR-183-5p group(P<0.05).Compared with the miR-183-5p+pcDNA3.1 group,the OD value,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group,whereas E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion was significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group(P<0.05).Overall,silencing miR-185-3p inhibited the growth of transplanted tumors and promoted FOXO1 expression.Conclusion Overexpression of miR-183-5p can inhibit apoptosis and promote the proliferation,migration,invasion,and EMT,of NSCLC cells by down-regulating FOXO1 expression.

Position of Village Regulations and Folk Conventions in the Villager Autonomy System

The Villagers' Charter of Self-government,as a senior form of village regulations and folk conventions,is the general charter of villagers' self-government. It is a small constitution of villagers,so it holds an important position in the villager autonomy system. Such position gives the credit to both historic and realistic objective factors. Rise of state power,vacancy of legislation,accumulation of excellent traditional resources,and democracy and contract spirit in village regulations and folk conventions will certainly accelerate development of village regulations and folk conventions and promote gradual improvement in the villager autonomy system.

Physiological roles of mitogen-activated-protein-kinase-activated p38-regulated/activated protein kinase

Mitogen-activated protein kinases(MAPKs)are a family of proteins that constitute signaling pathways involved in processes that control gene expression,cell division, cell survival,apoptosis,metabolism,differentiation and motility.The MAPK pathways can be divided into conventional and atypical MAPK pathways.The first group converts a signal into a cellular response through a relay of three consecutive phosphorylation events exerted by MAPK kinase kinases,MAPK kinase,and MAPK.Atypical MAPK pathways are not organized into this three-tiered cascade.MAPK that belongs to both conventional and atypical MAPK pathways can phosphorylate both non-protein kinase substrates and other protein kinases.The latter are referred to as MAPK-activated protein kinases.This review focuses on one such MAPK-activated protein kinase,MAPK-activated protein kinase 5(MK5)or p38-regulated/activated protein kinase(PRAK).This protein is highly conserved throughout the animal kingdom and seems to be the target of both conventional and atypical MAPK pathways.Recent findings on the regulation of the activity and subcellular localization,bona fide interaction partners and physiological roles of MK5/PRAK are discussed.

Long noncoding RNA negative regulator of antiviral response contributes to pancreatic ductal adenocarcinoma progression via targeting miR-299-3p

BACKGROUND Pancreatic ductal cancer(PDAC)has high malignancy and poor prognosis.Long noncoding RNAs(lncRNAs)are associated with high levels of malignancy,including PDAC.However,the biological and clinical significance of negative regulator of antiviral response(NRAV)in PDAC is unclear.AIM To study the regulatory role of lncRNA NRAV in PDAC.METHODS GEPIA analyzed lncRNA NRAV and miRNA(miR-299-3p)expression levels in PDAC tissues and measured them in PDAC cells by quantitative measurements in real time.The specific role of NRAV and miR-299-3p in cell proliferation and transfer potential was evaluated by cell formation analysis,Cell Counting Kit-8 and Transwell analysis.The relationship between NRAV and miR-299-3p was studied by predictive bioinformatics,RNA immunoassay,and fluorescence enzyme analysis.In vivo experiments included transplantation of simulated tumor cells under naked mice.RESULTS The expression level of lncRNA NRAV was higher in both tumor tissues and cell lines of PDAC and was negatively associated with the clinical survival of PDAC patients.Functionally,overexpression of NRAV promoted cell proliferation and metastasis of PDAC cells,while knockdown of NRAV reversed these effects.Finally,NRAV was performed as a molecular sponge of miR-299-3p.Moreover,overexpression of miR-299-3p could reverse the promoting effects of NRAV on cell proliferation and metastasis of PDAC cells.CONCLUSION NRAV facilitates progression of PDAC as a molecular sponge of miR-299-3p and may be a potential molecular marker for diagnosis and treatment of PDAC.

Synthesis,Crystal Structure and Plant Growth Regulatory Activity of 1-(3-Amino-[1,2,4]triazol-1-yl)-3,3-dimethyl-butan-2-one

The title compound 1-（3-amino-[1,2,4]triazol-1-yl）-3,3-dimethyl-butan-2-one（3） was synthesized by Hofmann-alkylation reaction of 1-chloro-3,3-dimethyl-butan-2-one（1） and ~1H-[1,2,4]triazol-3-ylamine（2） with equal amount of K_2CO_3 as acid acceptor. The structure of compound 3 was characterized by ~1H NMR, 13 C NMR, HRMS and single-crystal X-ray diffraction. The compound crystallizes in the monoclinic system, space group P21/n with a = 5.7227（8）, b = 27.924（4）, c = 6.2282（7） ？, β = 101.892（11）°, V = 973.9（2） ？~3, Z = 4, T = 180.00（10） K, μ（MoKα） = 0.087 mm^（-1）, Dc = 1.243 g/cm^3, 3832 reflections measured（3.648≤θ≤26.022°）, 1916 unique reflections（Rint = 0.0359, Rsigma = 0.0572） used in all calculations. The final R = 0.0557（I 〉 2σ（I）） and w R = 0.1276（all data）. Bioassay showed that 3 displayed excellent activity as plant growth regulator with inducing lateral root formation and enhancing primary root elongation at 0.27 mmol/L（50 ppm） in soybeen（He Feng-50）. Good water solubility was found with 50 mg in 1 m L of water. Therefore, application of 3 in agriculture is more environmentally friendly due to cosolvent-free condition, and results in improved abiotic-stress tolerance by affecting the root growth. And furthermore, it can be used as a precursor to investigate the function of regulating plant root growth.

circ_0003204 regulates the osteogenic differentiation of human adipose-derived stem cells via miR-370-3p/HDAC4 axis

Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.

A Novel Cellular Autoaggregative Developmentally CRP Regulated Behaviour Generates Massively Chondrule-Like Formations over Surface of Old <i>Escherichia coli</i>K-12 Macrocolony Biofilms

How Escherichia coli bacteria develop a particular colonial, 3-D biofilm morphological pattern is still a poorly understood process. Recently, we reported a new E. coli K-12 morphotype exhibited by old macrocolonies described as volcano-like. The formative developmental process of this morphotype has been presented as a suitable experimental model for the study of 3D patterning in macrocolony biofilms. Here, we report the optical microscopy observations and genetic analysis that have unveiled the existence of a novel autoaggregative behaviour which generates massive lumpiness over the surface of the volcano-like macrocolonies. These lumpy formations are generated by the autoaggregation and strong interaction of tightly packed bacterial cells in structures with a chondrule-like appearance which give the colony’s surface its characteristic microscopic lumpy phenotype. Furthermore, they exhibit different levels of maturation from the edge to the center of the colony. Hence, its generation appears to follow a spatiotemporal program of development during the macrocolony’s morphogenesis. Interestingly, the agar’s hardness influences the morphology exhibited by these formations, with high agar concentration (1.5%, 15 g/L) suppressing its development. This new auto-aggregative E. coli’s behaviour does not require the activity of the biofilm master regulator CsgD, the adhesiveness of flagella, pili type 1, adhesin Ag43, β-1,6-N-acetyl-D-glucosamine polymer-PGA, cellulose or colanic acid, but it is under glucose repression and the control of cAMP receptor protein (CRP). The possible physiological role of these chondrule-like formations in the adaptability of the colony to different stressful environmental conditions is discussed.

中国省域高等教育“放管服”改革政策的审视与优化

高等教育“放管服”改革是深化教育行政管理体制改革、转变政府职能的必由之路。以中国省域高等教育“放管服”改革政策文本为研究对象,应用内容分析方法,分别从简政放权、放管结合、优化服务三个方面对政策内容进行分析,将政策的典型特征提炼为:政策文本体例相似且均为联合发文、权力下放呈现一定梯度且强调试点先行、政策推进强化机制创新及营造良好环境。提出了不断完善政策体系、形成政策协同效应、强化政策执行合力等优化对策。

Ferroptosis mechanism and Alzheimer's disease

Regulated cell death is a genetically determined form of programmed cell death that commonly occurs during the development of living organisms.This process plays a crucial role in modulating homeostasis and is evolutionarily conserved across a diverse range of living organisms.Ferroptosis is a classic regulatory mode of cell death.Extensive studies of regulatory cell death in Alzheimer’s disease have yielded increasing evidence that fe rroptosis is closely related to the occurrence,development,and prognosis of Alzheimer’s disease.This review summarizes the molecular mechanisms of ferroptosis and recent research advances in the role of ferro ptosis in Alzheimer’s disease.Our findings are expected to serve as a theoretical and experimental foundation for clinical research and targeted therapy for Alzheimer’s disease.

规制的经济法属性及构成要素--《民法典》背景下的经济法理论变革

《民法典》背景下的经济法必须以尊重私权和私法自治为起点,但应从规制这一相反方向进行制度和理论构建。规制是以禁止、限制和强制方式限缩权利和自由的法律形式,源于并寓于经济法,由规制对象、目标、措施和机构等要素构成。作为一个枢纽型概念,规制集中反映了经济法的特质,并可将经济法与经济学及其他学科联结起来。规制是立法、行政和司法的共同行动,其源头是法律而非行政。行政规制的实质是执法,与规制完全不能等同。

父母自主支持与高中生学习投入:意向性自我调节和学校联结的作用

为探讨父母自主支持对高中生学习投入的影响及其内在作用机制,使用父母自主支持量表、意向性自我调节问卷、学校联结问卷和中文版学习投入量表对977名高中生进行施测。结果表明:(1)父母自主支持能够显著正向预测高中生的学习投入;(2)意向性自我调节在父母自主支持与学习投入关系中的中介作用显著;(3)父母自主支持影响学习投入的直接效应以及意向性自我调节中介作用的前半路径均受到学校联结的调节。研究结果凸显了家校协同在促进学生发展适应中的重要作用并揭示了其内在机制,对于推动家庭教育、提升学生学习投入和发展适应水平具有现实启示。

企业合规制度的建构逻辑:内力、外力与引力

企业合规的制度建设,发轫于企业内部的合规自治理需求,与此同时,企业外部法律责任的威慑力和市场声誉机制的约束持续向内部传动,不断改善企业治理方式和结构,增进企业合规治理的能动性。为克制企业内部自治无力与外部规制失焦形成的双向盲打和制度真空,国家务应提供规范化的制度导引,将内部合规治理的自治型制度融入国家对企业行为的规范制度体系,促成企业自治和法律规制的持续性互动以融合共治,制度化引导企业行为合规。由是,合规制度的完善重点在于:一是实体法上持续优化企业法律责任体系,程序法上继续完善刑事诉讼法、行政处罚法等以提供强规范;二是正确认识公司法的企业合规引导功能,从而在公司法体系内对企业提供内生性、整体性的软指引。

规则下乡何以促进村庄自主性——基于浙东张村的田野调查

增强村庄自主性是实现村庄治理能力现代化的有效途径。通过对浙东张村田野调查,基于规则下乡赋予的自主空间,进一步促进了村庄自主性,服务村庄有效完成了国家治理任务和回应群众诉求。规则下乡促进村庄自主性的形成,具体表现为创新执行上级实质性工作、灵活应对上级形式化工作、精准实施内生实质性工作和简约开展内生形式化工作。规则下乡促进村庄自主性的形成机制主要为:适度组织嵌入激活组织动员,推动村庄主动完成治理任务和回应群众诉求;灵活资源嵌入提升治理能力,赋予村庄有效增强治理能力和营造自主空间;科学考评嵌入推动政策执行,引导村庄科学配置治理资源和调试治理方式。

清代:中华民族命运共同体构建的重要历史时期--以清代基层社会组织重建和社会治理为视角的考察

以清代基层社会组织重建和社会治理为视角进行考察,清代是中华民族命运共同体构建的一个重要历史时期。清朝建立后,不仅继承了明朝的政治制度和国家治理体系,也继承了明朝的基层社会治理体系,重点对宗族、保甲、乡约以及会馆等民间组织的恢复与重建予以支持,并让渡部分公权力,使其统治触角向下延伸,“皇权”也由此渗入县以下的城乡基层社会。满族属于中华民族共同体的一员,而非域外之人,因此,他们一旦取得政权之后,除了保留满族的部分政治文化和风俗传统外,更多的是在大中华文化体系中寻找生存智慧与治国理政之道。在清代,不仅汉族、满族,也包括蒙古族、藏族或回族以及其他中国境内各民族,都主动或被动、自觉或不自觉地融入到中华民族命运共同体构建的历史进程之中。正因为如此,民族融会才升华为“中华民族”这一新概念。

裁量基准二阶构造的法理展开

现代社会的高阶立法主要是一种框架秩序,行政机关制定和适用裁量基准时享有二阶裁量空间,裁量基准兼具法制定与法适用的双重属性。但是,上位法规范和一般法原则仍对这种二阶裁量有框架拘束力,故行政裁量基准制度的统合法理是“行政自主与法律拘束的统一”。这一法理既是行政裁量之治的核心要义,也是司法监督执法裁量理由的基本准据。行政规章和规范性文件都是我国裁量基准的载体,但目前规范性文件趋同化程度较高,宜加强地方政府规章的立法表达,形成层级裁量基准体系。

Protein-spatiotemporal partition releasing gradient porous scaffolds and anti-inflammatory and antioxidant regulation remodel tissue engineered anisotropic meniscus

Meniscus is a wedge-shaped fibrocartilaginous tissue,playing important roles in maintaining joint stability and function.Meniscus injuries are difficult to heal and frequently progress into structural breakdown,which then leads to osteoarthritis.Regeneration of heterogeneous tissue engineering meniscus(TEM)continues to be a scientific and translational challenge.The morphology,tissue architecture,mechanical strength,and functional applications of the cultivated TEMs have not been able to meet clinical needs,which may due to the negligent attention on the importance of microenvironment in vitro and in vivo.Herein,we combined the 3D(three-dimensional)-printed gradient porous scaffolds,spatiotemporal partition release of growth factors,and anti-inflammatory and anti-oxidant microenvironment regulation of Ac2-26 peptide to prepare a versatile meniscus composite scaffold with heterogeneous bionic structures,excellent biomechanical properties and anti-inflammatory and anti-oxidant effects.By observing the results of cell activity and differentiation,and biomechanics under anti-inflammatory and anti-oxidant microenvironments in vitro,we explored the effects of anti-inflammatory and anti-oxidant microenvironments on construction of regional and functional heterogeneous TEM via the growth process regulation,with a view to cultivating a high-quality of TEM from bench to bedside.

lncRNACNN3-206 activates intestinal epithelial cell apoptosis and invasion by sponging miR-212, an implication for Crohn’s disease

BACKGROUND Statistics indicate that the incidence of Crohn’s disease(CD)is rising in many countries.The poor understanding on the pathological mechanism has limited the development of effective therapy against this disease.Previous studies showed that long noncoding RNAs(lncRNAs)could be involved in autoimmune diseases including CD,but the detailed molecular mechanisms remain unclear.AIM To identify the differentially expressed lncRNAs in the intestinal mucosa associated with CD,and to characterize their pathogenic role(s)and related mechanisms.METHODS The differential expression of lncRNAs was screened by high-throughput RNA sequencing,and the top candidate genes were validated in an expanded cohort by real-time PCR.The regulatory network was predicted by bioinformatic software and competitive endogenous RNA analysis,and was characterized in Caco-2 and HT-29 cell culture using methods of cell transfection,real-time PCR,Western blotting analysis,flow cytometry,and cell migration and invasion assays.Finally,these findings were confirmed in vivo using a CD animal model.RESULTS The 3'end of lncRNACNN3-206 and the 3’UTR of Caspase10 contain highaffinity miR212 binding sites.lncRNACNN3-206 expression was found to be significantly increased in intestinal lesions of CD patients.Activation of the lncRNACNN3-206-miR-212-Caspase10 regulatory network led to increased apoptosis,migration and invasion in intestinal epithelial cells.Knockdown of lncRNACNN3-206 expression alleviated intestinal mucosal inflammation and tissue damage in the CD mouse model.CONCLUSION lncRNACNN3-206 may play a key role in CD pathogenesis.lncRNACNN3-206 could be a therapeutic target for CD treatment.

职业放贷行为的认定与效力--基于122份判决的实证考察

职业放贷行为不同于一般民间借贷,涉嫌危害金融秩序与安全,故司法应平衡好“自治”和“管制”的双重目标,优化对职业放贷行为的司法规制。一方面,在职业放贷行为认定中,应将形式审查与实质审查相结合,严格认定职业放贷行为,以维护民事主体的意思自治;另一方面,在认定职业放贷行为后,可依《民法典》第153条,以规范目的探寻为进路,通过利益衡量判定职业放贷行为无效;并依《民法典》第157条来具体确定职业放贷行为无效的法律后果,否定职业放贷人利息请求权和资金占用使用费请求权,严格限定担保人的担保责任,最终实现“管制”目的,维护金融安全。

软法视野下村规运用实证考察与机理重塑--以构建情理法统一为中心视角展开

村规蕴含的公序良俗,不仅是《民法通则》的基本原则,也是"民间法"软治理的规范之一。村规在乡土社会发挥着重要的治理功能。然而现实却存在着大量异化的村规,其根源在于村规的制定缺失民意基础、制定程序随意。在我国法治建设进程中,制定法应理性引导村规条约制定、审查与备案,预留村规合理的规制空间;反之,村规也应恪守法治思维,体现基层民众真实意愿。为制定法与村规在乡土治理领域寻求一条协同共存之路。

博弈与权衡:智能教育算法的规制性与人的自主性

在智能教育中,算法的规制性能够辅助人做出科学的决策,而人的自主性又能够对教学情境做出自主的感知判断,两者各有所长。然而,在具体教学实践中常出现两种偏向:一是过度青睐算法的规制性,这种偏向存在限制人的自主性发展的嫌疑;二是过度强调人的自主性发展,这种偏向又存在遮蔽算法工具理性、影响教育效益的可能。为此,文章首先对算法的规制性与人的自主性展开分析,并提出智能教育教学应在动态博弈中权衡算法的规制性与人的自主性;然后文章从目的、过程及结果三层面提出算法的规制性与人的自主性的耦合方案,即在精准挖掘教育需求中促进人的自主性发展,在人机协同过程中合理设计算法的规制性,在厘清运用边界中实现教育价值旨归,旨在理性回归智能时代的教育本质,为算法在教育中的合理与向善发展提供有效路径与方法。