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关键词： 医德   细胞生物学   实验课   实验室   课程思政   安全教育  

摘要： 医学教育的主要目的是培养医术精湛、医德高尚的医务人员。思政教育需要载体,驱动医学教育课程思政落地,以实验课作为专业教育与思政教育的整合途径,深入挖掘思政元素,融入集体主义精神、生命价值观、责任心、同理心和安全意识等教育,以规范化操作训练、视频、图片和参与管理等方式实施,学生的实验习惯和道德品质不断提升。

关键词： 细胞生物学实验   教学改革   评价体系  

摘要： 目前,细胞生物学实验教学的主要问题在于实验课内容设置没有与时俱进,教学手段和方式比较僵化,评价体系比较单一。针对这几个问题,该文进行了一些探索,增加了综合设计实验占比;引入多样化教学手段,将传统讲授与多媒体相结合;改变考核方式,重视过程评价;通过这些改革,希望形成以学生为中心设计、开展实验教学的新教学理念,和基于学生全面发展的创新性实验教学模式,培养和提高学生的实验设计能力、实验动手能力、理论联系实际能力、探索创新能力。

关键词： 生物物理学   学术期刊   系统生物学   神经科学   细胞生物学   栏目设置   学科范围   基因组学  

摘要： (2020年12月修订)1征稿范围与栏目设置1.1征稿范围本刊为国内外公开发行的全国性学术期刊,月刊,每月20日发行.征稿的学科范围是生物化学、分子生物学、生物物理学、细胞生物学、神经科学、免疫学,以及基因组学、蛋白质组学、系统生物学等相关学科领域.

关键词： 高迁移率族蛋白1   胶质瘤   生物学功能  

摘要： 目的:检测高迁移率族蛋白1(high mobility group box-1,HMGB1)在胶质瘤组织和细胞中的表达情况,了解HMGB1对胶质瘤细胞增殖、侵袭迁移以及细胞周期的影响。方法:将HMGB1 siRNA真核表达质粒在脂质体介导下转染胶质瘤细胞U373、U87(HMGB1 siRNA组),以转染无关序列siRNA质粒的细胞(negative control,NC)和未转染的胶质瘤细胞(Control,Con)作为对照。采用RT-PCR和Western blot检测HMGB1 mRNA和蛋白表达,通过CCK-8实验、Transwell实验和细胞划痕实验分别检测胶质瘤细胞增殖、侵袭和迁移能力,用流式细胞仪检测胶质瘤细胞周期的变化。结果:HMGB1 mRNA在胶质瘤组织和细胞水平上都较非肿瘤性脑组织和正常胶质细胞明显高表达,差异有统计学意义(P<0.05),CCK-8和划痕实验提示HMGB1 siRNA组细胞增殖和迁移能力受到明显抑制(P<0.05),Transwell实验显示HMGB1 siRNA组穿膜侵袭到基质胶的细胞数明显低于Con组和NC组,细胞侵袭能力减弱(P<0.05),流式细胞分析提示HMGB1 siRNA组位于S期细胞比例较Con和NC组明显减少(P<0.05),提示细胞周期发生S期阻滞。结论:应用RNAi能有效抑制HMGB1基因表达,HMGB1低表达后能抑制胶质瘤细胞增殖、侵袭和迁移,并使得细胞周期中进入S期的细胞比例明显减少,细胞周期发生阻滞,这可能为胶质瘤的基因治疗提供了新思路。

关键词： Chronic stress   Social isolation   Anxiety- and depressive-like behaviour   Systemic inflammation   Neuroinflammation   Hippocampal neurogenesis  

摘要： Unpredictable chronic mild stress (UCMS) is one of the most commonly used, robust and translatable models for studying the neurobiological basis of major depression. Although the model currently has multiple advantages, it does not entirely follow the trajectory of the disorder, whereby depressive symptomology can often present months after exposure to stress. Furthermore, patients with depression are more likely to withdraw in response to their stressful experience, or as a symptom of their depression, and, in turn, this withdrawal/isolation can further exacerbate the stressful experience and the depressive symptomology. Therefore, we investigated the effect(s) of 6 weeks of UCMS followed by another 6 weeks of social isolation (referred to as UCMSI), on behaviour, corticosterone stress responsivity, immune system functioning, and hippocampal neurogenesis, in young adult male mice. We found that UCMSI induced several behavioural changes resembling depression but did not induce peripheral inflammation. However, UCMSI animals showed increased microglial activation in the ventral dentate gyrus (DG) of the hippocampus and astrocyte activation in both the dorsal and ventral DG, with increased GFAP-positive cell immunoreactivity, GFAP-positive cell hypertrophy and process extension, and increased s100 beta-positive cell density. Moreover, UCMSI animals had significantly reduced neurogenesis in the DG and reduced levels of peripheral vascular endothelial growth factor (VEGF) - a trophic factor produced by astrocytes and that stimulates neurogenesis. Finally, UCMSI mice also had normal baseline corticosterone levels but a smaller increase in corticosterone following acute stress, that is, the Porsolt Swim Test. Our work gives clinically relevant insights into the role that microglial and astrocyte functioning, and hippocampal neurogenesis may play in the context of stress, social isolation and depression, offering a potentially new avenue for therapeutic target.

关键词： MiR-20a-5p   miR-132   Novel miRs   Cardiac fibrosis   Autophagy   Reactive oxygen species  

摘要： Background: Myocardial fibrosis is a hallmark of the failing heart, contributing to the most common causes of deaths worldwide. Several microRNAs (miRNAs, miRs) controlling cardiac fibrosis were identified in recent years;however, a more global approach to identify miRNAs involved in fibrosis is missing. Methods and results: Functional miRNA mimic library screens were applied in human cardiac fibroblasts (HCFs) to identify annotated miRNAs inducing proliferation. In parallel, miRNA deep sequencing was performed after subjecting HCFs to proliferating and resting stimuli, additionally enabling discovery of novel miRNAs. In-depth in vitro analysis confirmed the pro-fibrotic nature of selected, highly conserved miRNAs miR-20a-5p and miR-1323p. To determine downstream cellular pathways and their role in the fibrotic response, targets of the annotated miRNA candidates were modulated by synthetic siRNA. We here provide evidence that repression of autophagy and detoxification of reactive oxygen species by miR-20a-5p and miR-132-3p explain some of their pro-fibrotic nature on a mechanistic level. Conclusion: We here identified both miR-20a-5p and miR-132-3p as crucial regulators of fibrotic pathways in an in vitro model of human cardiac fibroblast biology.

关键词： 肝癌   预后模型   肽基脯氨酰顺反异构酶基因   FKBP1A蛋白   细胞生物学  

摘要： 目的:　　1.构建基于肽基脯氨酰顺反异构酶(PPIase)基因特征的肝癌预后模型,探讨该模型在肝癌中的预后价值。　　2.研究FKBP1A在肝癌中的临床意义以及探讨其在肝癌细胞中的生物学作用。　　3.初步探讨FKBP1A调控肝癌细胞生物学功能的作用机制。　　方法:　　1.从TCGA和ICGC数据库下载肝癌患者的转录组测序数据和临床数据分别作为训练集和验证集。从分子特征数据库获取PPIase基因集。应用R软件对训练集中的肝癌和癌旁组织进行PPIase基因的差异表达分析，筛选出肝癌中差异表达的基因。采用Cox、Lasso回归等方法构建基于PPIase基因特征的预后模型。Kaplan-Meier生存分析评价模型的预后价值，并用独立的外部数据集对模型进行验证。最后，联合临床分期与预后风险评分绘制列线图和校准曲线。　　2.应用生物信息学方法在TCGA、GEPIA、HPA数据库分析FKBP1A在肝癌中的表达以及临床意义。应用western blot技术检测FKBP1A蛋白在肝癌和癌旁组织中的表达。采用siRNA技术敲低Huh7和HepG2细胞中FKBP1A基因的表达水平。应用CCK-8法检测肝癌细胞的增殖能力;流式细胞仪检测肝癌细胞的凋亡情况;Transwell检测肝癌细胞的迁移能力。GSEA富集分析探索与FKBP1A可能相关的KEGG通路，western blot技术检测相关信号通路分子的表达情况，探索其调控机制。　　结果:　　1.肝癌及癌旁组织的基因差异表达分析显示有16个表达上调的PPIase基因，下调的基因0个。通过Cox、Lasso和逐步回归分析最终得到基于3个基因特征（FKBP6，CWC27，FKBP1A）的肝癌预后模型。Kaplan-Meier曲线显示高风险评分组的肝癌患者预后更差（Plt;0.05）。ROC曲线显示，预测1、2、3年生存率的AUC值分别为0.725、0.680、0.644。此外，验证集对模型的评价结果与训练集的基本一致，并成功构建了整合TNM分期和预后风险评分相结合的列线图，校准曲线与对角线的匹配度较高。　　2.基于TCGA、GEPIA和HPA数据库的生物信息分析发现，与癌旁组织和正常肝组织比较，肝癌组织中FKBP1A mRNA和蛋白水平呈现高表达。本研究收集了23对肝癌和癌旁组织进行western blot检测，结果显示FKBP1A蛋白在肝癌组织中的表达显著高于癌旁组织（Plt;0.05）。来源于TCGA和GEPIA的数据显示，肝癌的病理分级和分期越高，FKBP1A mRNA的表达越高。Kaplan-Meier曲线显示高FKBP1A mRNA表达组的肝癌患者的总生存期和无病生存期更差（Plt;0.05）。CCK8和流式细胞仪结果显示敲低FKBP1A基因对Huh7和HepG2细胞的增殖和凋亡没有显著影响。Transwell实验结果显示敲低FKBP1A基因显著抑制了Huh7和HepG2细胞的迁移。GSEA富集分析显示，MAPK、NOTCH、WNT信号通路在FKBP1A高表达组中显著富集。敲低FKBP1A基因后，Huh7和HepG2细胞中β-catenin蛋白表达下调。　　结论:　　1.基于3个PPIase基因特征的肝癌预后模型具有良好的预后价值，有希望为肝癌患者的预后风险分层提供参考。　　***1A是肝癌患者独立的不良预后因子，并且在肝癌中可能起促癌的作用，有望作为肝癌治疗的潜在靶点以及预后的生物标志物。　　***1A可能通过激活WNT信号通路促进肝癌细胞的迁移。

关键词： Cellular biology   Molecular biology   Microbiology  

摘要： Methane levels have rapidly increased in concentration in the atmosphere due to anthropogenic inputs making it a crucial abatement target against global climate change. The aerobic methanotrophs are bacteria capable of growth solely on methane as substrate and are genetically tractable and amenable to large scale fermentation making them ideal model system for the engineering of C1- biocatalysts. In this dissertation I examine the effects of trace minerals and substrate on the cell performance of Methylotuvimicrobium alcaliphilum 20ZR by contextualizing multi-omics with structural characterization through electron microscopy. This reveals the development of the intracytoplasmic membrane network within the cell which is affected by metals and leads to the increased gas consumption. Occupying the cell volume with membranes leads to an overflow metabolic state in conditions with sufficient substrate and oxygen in which fermentation products such as formate are excreted.. The nature of structural features revealed by microscopy is explored to determine the genetic elements for expression and transport of the surface layer protein. Genes responsible for the large and small proteins of the surface layer protein matrix are identified and a Type 1 Secretion System is explored as the potential mechanism of export for this cell surface feature. Concluding with an exploration of a novel methane chemotactic phenotype, which was identified by structural modelling and confirmed through a methane-based chemotaxis assay. This work demonstrates the importance of tying systems-level data such as proteomics with structural information gathered by electron microscopy to explain and explore phenotypes of performance such as growth rate and substrate consumption to cell biology of a model organism used as a methane-based biotechnology strain.

关键词： 细胞生物学   开放性   实验教学   教学模式  

摘要： 细胞生物学是生物相关学科的基础,是高等学校生物科学相关专业学生必修的专业课程,在生物科学相关的各种专业人才的培养中具有非常重要的作用。随着当今社会对应用型人才需求的不断增长,传统的实验教学方式已经越来越不能适应现代社会的要求。本文对开放性实验教学模式的开放性表现以及它在应用型人才培养方面的意义和困难进行了阐述,并分析了开展半开放性实验教学的必要性和优势。

关键词： THBS1   乳腺癌   增殖   凋亡   侵袭  

摘要： 研究背景:乳腺癌是危害女性健康最常见的恶性肿瘤之一,其发病率在全球范围呈逐年上升趋势。临床治疗乳腺癌主要采取手术切除及术后化疗,但由于侵袭性强、复发率高的特点,乳腺癌的病死率位居女性恶性肿瘤之首。血小板凝血酶蛋白-1(thrombospondin-1,THBS1)是由多种细胞合成分泌的一种多功能糖蛋白,在肿瘤中的异常表达有致癌或抑癌功能,在肿瘤的发展过程中起到较为复杂的调控作用,其对乳腺癌进展过程起到的调控作用尚不明确。目的:1.分析THBS1m RNA在乳腺癌中的表达水平及其与年龄、性别、TNM分期、病理分期、预后等临床病理特征的相关性。2.探讨小干扰RNA(small interfering RNA,si RNA)介导的THBS1基因沉默对人乳腺癌MCF-7及MDA-MB-231细胞增殖、侵袭及凋亡的影响。方法:1.生物信息学分析:利用TCGA数据库及GEPIA数据库分析THBS1m RNA在乳腺癌中的表达情况,分析THBS1的表达与年龄、性别、TNM分期等临床病理特征是否存在相关性。2.研究利用si RNA介导的THBS1基因沉默对乳腺癌MCF-7及MDA-MB-231细胞生物学行为的影响:(1)通过脂质体转染法,将三条不同序列的特异性针对THBS1基因的si RNA(简称si RNA-THBS1)转染至MCF-7及MDA-MB-231细胞中。(2)通过实时荧光定量PCR技术(q RT-PCR)检测三个不同序列的si RNA-THBS1对THBS1的沉默效果,并筛选出沉默效果最佳的序列,后续实验均采用转染该序列si RNA进行THBS1基因沉默。(3)Cell Counting Kit-8(CCK-8)增殖实验检测THBS1基因沉默对乳腺癌细胞增殖能力的影响。(4)Transwell小室基质侵袭实验检测THBS1基因沉默对乳腺癌细胞侵袭能力的影响。(5)流式细胞学技术Annexin V-FITC/PI双染法检测THBS1基因沉默对乳腺癌细胞凋亡的影响。所有实验均重复三次,实验结果使用SPSS23.0统计软件进行统计学分析,P<0.05认为差异有统计学意义。结果:1.生物信息学分析:THBS1在乳腺癌组织中的表达高于正常组织(P<0.05);在乳腺癌中,THBS1的表达与年龄、性别、病理分期、T(肿瘤大小)、M(远处转移)均无关,(P>0.05);与N(淋巴结转移)有关,(P<0.05)。2.细胞实验:(1)q RT-PCR:三种序列的si RNA-THBS1均可沉默THBS1,其中序列-1(si RNA-THBS1-1)的沉默效率最高,该序列si RNA对MDA-MB-231细胞的沉默效率达到(62.9±2.9)%,对MCF-7细胞的沉默效率达到(94.98±0.37)%。(2)CCK-8细胞增殖实验:沉默THBS1基因后,MCF-7细胞的增殖能力较空白对照组、si RNA-NC组均明显降低(P<0.05),MDA-MB-231细胞的增殖能力较其他对照组无明差异(P>0.05)。(3)Transwell小室基质侵袭实验:经苏木素染色计数后发现,两种细胞的si RNA-THBS1组侵袭细胞数均少于si RNA-NC组(P<0.05),差异具有统计学意义。(4)流式细胞学技术检测细胞凋亡:两种细胞si RNA-THBS1组凋亡率均高于si RNA-NC组,P<0.05,差异具有统计学意义。结论***1在乳腺癌中表达高于正常组织,且THBS1的表达与淋巴结转移相关;***1基因促进乳腺癌细胞MCF-7细胞的增殖,促进MDA-MB-231及MCF-7细胞的侵袭,并抑制两种细胞凋亡。