Hsa-miR-5195-3p对人宫颈癌细胞SiHa增殖、迁移与侵袭的影响

目的:探讨mi R-5195-3p对人宫颈癌细胞系Si Ha增殖、迁移与侵袭的影响。方法:采用qRT-PCR检测人宫颈癌细胞SiHa和正常上皮细胞HaCaT中mi R-5195-3p的表达水平。将mi R-5195-3p mimic转染至Si Ha细胞中构建外源性过表达细胞株,阴性对照组中则转染NC mimic,并用q RT-PCR验证转染效率;通过MTT和集落形成实验检测细胞增殖能力;划痕愈合实验检测细胞横向迁移能力; Transwell小室实验检测细胞纵向迁移能力和侵袭能力;采用qRT-PCR和Western blot检测E-cadherin、Vimentin与snail m RNA转录水平及蛋白表达水平。结果:宫颈癌细胞Si Ha中的mi R-5195-3p表达水平较HaCaT偏低(P 0. 05)。与阴性对照组相比,转染mi R-5195-3p mimic的SiHa细胞中mi R-5195-3p水平显著增高(P 0. 01);并且其体外增殖(P 0. 001),迁移(P 0. 001)与侵袭能力(P 0. 001)明显减弱;同时E-cadherin表达水平上调而Vimentin、snail表达水平下调。结论:过表达mi R-5195-3p可能通过阻碍EMT通路抑制宫颈癌细胞Si Ha的增殖,迁移与侵袭。     

miR-223在结肠癌组织中的表达及促进结肠癌细胞迁移侵袭的机制研究

目的:探讨微小RNA-223 (mi R-223)在结肠癌组织中的表达及对结肠癌HT-29细胞侵袭、迁移能力的影响及机制。方法:检测mi R-223在结肠癌组织与癌旁组织中的表达。通过脂质体转染法将mi R-223模拟物(mi R-223 mimics,mi R-223 mimics组)及microRNA无关序列(mi R-223 NC,NC组)转染入结肠癌HT-29细胞。采用Real-time PCR检测转染后细胞中mi R-223和TWIST的表达,Western blot检测TWIST的蛋白表达,Tranwell检测细胞的迁移与侵袭能力。双荧光素酶报告基因检测mi R-223对TWIST基因启动子活性的影响。采用Transwell迁移与侵袭实验检测mi R-223 mimic及Twist si RNA共转染后人结肠癌细胞系HT-29迁移与侵袭能力的变化。结果:与癌旁结肠组织比较,mi R-223在结肠癌组织中呈现明显高表达(P0.05);与空白对照组和mi R-223 NC组比较,转染mi R-223 mimics后的HT-29细胞中的mi R-223表达显著增加(P0.05)。与阴性对照组和空载转染组相比较,mi R-223 mimics转染组穿透的细胞数目明显增加(P0.05),且mi R-223 mimics转染组的细胞侵袭能力显著增强(P0.05)。与mi R-223 NC组和空白对照组比较,转染mi R-223 mimics的HT-29细胞的TWIST基因m RNA和蛋白表达均显著增加(P0.05)。双荧光素酶检验结果显示TWIST为mi R-223的下游靶基因。共转染TWIST si RNA和mi R-223 mimics的结肠癌HT-29细胞的迁移与侵袭能力较单独转染mi R-223 mimics的HT-29细胞显著减弱(P0.05)。结论:mi R-223可能通过上调下游靶基因TWIST水平促进结肠癌HT-29细胞的迁移与侵袭。     

miR-345靶向调控 TGM1抑制膀胱癌的初步研究

目的:探讨mi R-345调控TGM1表达影响膀胱癌的分子生物学机制。方法:首先,采用RT-qPCR检测T24和RT4细胞中mi R-345、TGM1的表达;再采用mi RNA-NC、mi R-345 mimic、NC inhibitor、mi R-345 inhibitor、control si RNA、si TGM1和pc-DNA3.1/TGM1等转染膀胱癌细胞;然后,采用MTT实验检测细胞增殖,Transwell实验检测细胞侵袭,流式细胞仪检测细胞凋亡,双荧光报告酶检测mi R-345的靶基因;最后,采用Western blot检测TGM1在细胞中的表达。结果:mi R-345在T24和RT4细胞中表达低于正常细胞(P0.05)。mi R-345过表达时,T24和RT4细胞的增殖侵袭能力减弱,细胞凋亡率上升;mi R-345表达沉默时,细胞增殖和侵袭能力增强,细胞凋亡率下降。双荧光报告基因检测结果显示TGM1为mi R-345的靶基因,mi R-345过表达抑制TGM1的表达(P0.05);mi R-345表达沉默时则表达上调(P0.05)。当TGM1表达沉默时,T24和RT4细胞的增殖和侵袭能力减弱,细胞凋亡率上升;TGM1过表达时该细胞的增殖和侵袭能力增强,细胞凋亡率下降。结论:mi R-345通过下调靶基因TGM1的表达,抑制膀胱癌细胞的增殖、侵袭并促进细胞凋亡。     

RNA干扰HIF-1α影响宫颈癌HeLa细胞上皮细胞间质变标记蛋白的研究

目的 通过RNA干扰技术沉默人宫颈癌HeLa细胞缺氧诱导因子-1α（hypoxia inducible factor-1α,HIF-1α）的表达,在细胞水平研究HIF-1α对宫颈癌HeLa细胞上皮细胞-间质变标记蛋白（E-cadherin,N-cadherin,vimentin）表达的影响.方法 将人宫颈癌HeLa细胞株（H0细胞）、转染pGenesil-1空白质粒的HeLa细胞株（H1细胞）及转染HIF-1α-shRNA质粒的HeLa细胞株（H2细胞）分别行常氧及缺氧（150 μmol/L CoCl2培养12 h）培养,应用Western印迹、免疫细胞化学法检测每组HeLa细胞中HIF-1α、上皮标记蛋白E-cadherin、间质标记蛋白vimentin、N-cadherin的表达情况.结果 Western印迹分析各缺氧组HIF-1α、E-cadherin、vimentin及N-cadherin的平均光密度值,免疫细胞化学显示H0细胞、H1细胞缺氧时HIF-1α、N-cadherin、vimentin蛋白高表达,而E-cadherin低表达,H2细胞在乏氧时HIF-1α、N-cadherin、vimentin蛋白表达显著减弱,而E-cadherin高表达.结论 通过shRNA干扰抑制人宫颈癌HeLa细胞HIF-1α,可一定程度上抑制乏氧环境中宫颈癌HeLa细胞上皮细胞-间质变.     

DZNep通过上调miR-200c表达延缓MGC-803胃癌细胞侵袭和迁移过程

本文旨在研究组蛋白甲基化修饰调控对胃癌细胞mi R-200c的表达调节以及对癌细胞的侵袭和迁移的作用。组蛋白甲基转移酶抑制剂DZNep(2.5μmol/L)处理MGC-803胃癌细胞系,用实时定量PCR(q RT-PCR)检测细胞mi R-200c的表达变化,用Western blot检测上皮间质转化相关蛋白、EZH2、EED、SUZ12、H3K27me3及MMP9的蛋白表达变化,用细胞划痕实验和Transwell法检测细胞迁移和侵袭。结果显示,与对照(DMSO处理)组相比,DZNep(2.5μmol/L)处理的MGC-803肿瘤细胞mi R-200c基因的表达显著提高,ZEB1、ZEB2、N-cadherin的表达显著下调,E-cadherin的表达上调,EZH2、EED、SUZ12、H3K27me3及MMP9的表达均显著降低,细胞迁移、侵袭能力均减弱。以上结果提示,DZNep通过上调mi R-200c的表达延缓胃癌细胞侵袭、迁移过程,其机制涉及对上皮间质转化相关蛋白和PRC2(polycomb repressive complex 2)的表达调节。     

氧化应激诱导内皮细胞凋亡microRNA表达改变的研究

目的:研究氧化应激诱导的内皮细胞micro RNA的表达变化。方法:ECM(Endothelial Cell Medium)培养人脐静脉内皮细胞,利用不同浓度双氧水(0μmol/L,200μmol/L,500μmol/L,800μmol/L)刺激24小时后应用流式细胞术检测其凋亡水平。提取细胞总RNA,利用实时定量PCR(Quantitive real-time PCR;q RT-PCR)检测micro RNA表达量变化,并利用生物信息学软件预测可能的靶基因。结果:加入不同浓度双氧水处理24 h后的内皮细胞总凋亡率均显著高于对照组,200μmol/L、500μmol/L和800μmol/L组的凋亡率分别为(13.31%vs 4.75%,35.9%vs 4.75%,89.75%vs 4.75%,P0.01)。200μmol/L的双氧水处理内皮细胞后,micro RNA的表达出现了明显的改变。其中mi R-92a、mi R-126的表达明显下调(P0.05),mi R-181a、mi R-217、mi R-34a和mi R-320的表达明显上调(P0.05)。靶基因预测显示mi R-320、mi R-92a可能调控多个和内皮细胞凋亡相关的基因表达。结论:在氧化应激诱导的内皮细胞凋亡中,mi RNA表达发生改变并可能参与调控内皮细胞功能。     

miR-181a通过调控上皮间质转化过程影响卵巢癌细胞的迁移和侵袭

目的:探讨mi R-181a在卵巢癌细胞中的表达及对卵巢癌细胞的迁移和侵袭的影响和可能机制。方法:采用细胞免疫荧光检测卵巢癌细胞中抗波形蛋白和E-钙粘蛋白的表达,Western blotting检测mi R-181a对抗波形蛋白和E-钙粘蛋白的表达情况的调控;划痕愈合实验检测mi R-181a对卵巢癌细胞迁移能力的影响;Transwell侵袭实验检测mi R-181a对卵巢癌细胞侵袭能力的影响。结果:增加mi R-181a的表达后,EMT过程中的相关蛋白激活水平下调,mi R-181a一定程度上可以抑制卵巢癌细胞的EMT过程;过表达mi R-181a后可以明显影响Vimentin[D1(4.58±0.85)vs(0.29±0.02),P0.05;D5(4.16±0.79)vs(0.29±0.02),P0.05]和E-Cadherin[D1(4.75±0.41) vs.(4.56±0.38),P0.05;D5(2.19±0.18) vs.(4.56±0.38),P0.05]蛋白的表达;COC1细胞的迁移能力随mi R-181a的表达的增高而降低[(19.24±4.31)%vs.(25.95±6.02)%,P0.05;(51.25±8.75)%vs.(73.49±12.54)%,P0.05];过表达mi R-181a后可以一定程度上减弱卵巢癌COC1细胞的侵袭能力[(74.64±8.21)vs(231.98±21.72),P0.05]。结论:mi R-181a通过调控上皮间质转化过程影响卵巢癌细胞的迁移和侵袭行为。     

MiR-301b抑制转录因子Klf4影响肝癌细胞迁移

目的:探讨mi R-301b对肝癌细胞迁移能力的影响及其分子机制,为肝癌的分子靶向治疗研究提供新线索。方法:体外培养人肝癌细胞株SK-Hep-1、HCC-LM3和人永生化肝细胞株L02,采用RT-PCR方法检测mi R-301b表达。通过生物信息学软件Targetscan及mi Randa预测mi R-301b的靶基因,筛选出转录因子Klf4基因为mi R-301b的下游靶基因,通过双荧光素酶报告基因实验和Western Blot实验证明其调控作用。通过划痕和Transwell实验探究mi R-301b靶向Klf4基因对肝癌细胞迁移性的影响,Western Blot检测mi R-301b对上皮间质转化标记物E-cadherin、N-cadherin蛋白表达的影响。结果:与正常肝细胞相比,肝癌细胞株中mi R-301b表达水平明显升高。瞬时转染mi R-301b mimic后,实验组mi R-301b的表达显著高于对照组;瞬时转染mi R-301b inhibitor后,实验组mi R-301b的表达显著低于对照组。双荧光素酶报告基因实验显示:mi R-301b直接作用于Klf4基因的3'UTR区,并下调Klf4蛋白的表达,与软件预测结果相符合。划痕实验及Transwell迁移实验显示:mi R-301b通过下调Klf4基因,促进肝癌细胞的迁移。进一步实验显示:过表达mi R-301b显著下调E-cadherin的表达,而上调N-cadherin的表达。结论:mi R-301b在肝癌细胞SK-Hep-1、HCC-LM3中高表达,可能通过抑制靶基因Klf4的表达,促进肝癌的迁移,mi R-301b可能参与了肝癌细胞的上皮间质转化过程。     

miR-10a对类风湿性关节炎滑膜细胞侵袭、迁移能力的影响及机制

目的:以类风湿性关节炎(RA)患者滑膜细胞为研究对象,明确mi R-10a对滑膜细胞侵袭、迁移能力的影响及机制。方法:首先用q RT-PCR对筛选得到的异常表达mi RNA进行验证,进而利用生物信息学分析结合报告基因、Western印迹方法,明确mi R-10a的靶基因,最后采用Transwell、划痕实验考察mi R-10a对RA成纤维细胞样滑膜细胞(FLS细胞)侵袭能力的影响。结果:mi R-10a在RA患者滑膜组织及细胞中低表达;TAK1和BTRC是mi R-10a的靶基因;mi R-10a可促进IL-6、IL-8等炎症因子的表达;mi R-10a可促进FLS细胞的侵袭、迁移。结论:mi R-10a可通过调节NF-κB的活性影响RA FLS细胞的侵袭、迁移。     

miR-24对心脏成纤维细胞生长和迁移的影响及机制研究

目的:探讨miR-24对心脏成纤维细胞的生长和迁移的影响及机制。方法:采用RT-PCR检测心肌细胞和心脏成纤维细胞中mi R-24的表达水平。在心脏成纤维细胞中转染mi R-24 mimics、mimics control、mi R-24 inhibitors、inhibitors control后,通过Western blot检测细胞中Col-1、α-SMA的表达,MTT检测细胞增殖情况,流式细胞仪检测细胞凋亡情况,Transwell小室检测细胞迁移能力。通过靶基因预测库预测mi R-24的靶基因,荧光素酶鉴定靶基因的正确性,并通过RT-PCR和Western blot检测转染后细胞中Furin的表达。结果:心脏成纤维细胞HEH2中mi R-24的表达水平与心肌细胞H9C2相比差异显著(P0.01),心脏成纤维细胞中mi R-24表达上调。mi R-24 mimics组中Col-1、α-SMA的表达水平明显低于mimics control组(P0.01)。mi R-24 mimics组中细胞OD值较mimics control组显著降低(P0.01),mi R-24 inhibitors组中细胞OD值较inhibitors control组显著升高(P0.01)。mi R-24 mimics组、mimics control组、mi R-24 inhibitors组、inhibitors control组细胞凋亡无显著性差异(P0.05)。mi R-24 mimics组细胞迁移数目明显低于mimics control组,差异显著(P0.01),mi R-24 inhibitors组细胞迁移数目高于inhibitors control组,差异显著(P0.05)。mi R-24 mimics组细胞中Furin蛋白和m RNA水平明显降低,野生型Furin和mi R-24 mimics共转染的细胞中荧光素酶活性最低。结论:mi R-24在心脏成纤维细胞中高表达,通过靶基因Furin抑制心脏成纤维细胞合成Col-1、α-SMA,抑制心脏成纤维细胞增殖和迁移。     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

鸡传染性法氏囊病病毒研究进展

传染性法氏囊病(Infection bursal disease, IBD)是由鸡传染性法氏囊病毒(Infectious bursal disease virus, IBDV)引起的鸡和火鸡的一种高度接触性传染病,给世界各国的禽养殖业带来了巨大损失.自IBDV发现至今新的变异株不断出现,分子结构的改变导致病毒致病力的改变及宿主对疫苗应答的改变,使得传统的疫苗已不能控制其流行,因此各国学者对其基因组结构和功能进行了广泛深入的研究,并积极研制新型有效的疫苗以达到防治的目的.     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).