CD47基因对食道癌细胞增殖的影响

本研究将CD47-si RNA转染至食道癌细胞,采用蛋白免疫印迹检测食道癌细胞中CD47蛋白的表达,MTT法检测CD47-siRNA转染组和空质粒转染组(对照组)食道癌细胞增殖状态,蛋白免疫印迹检测CD47-siRNA转染组和空质粒转染组(对照组)食道癌细胞中PCNA蛋白表达,DCFDA染色流式细胞仪检测食道癌细胞CD47-siRNA转染组和空质粒转染组(对照组)中ROS水平,以探究CD47基因对食道癌发生发展的影响。研究结果表明,CD47-si RNA转染组食道癌细胞中CD47蛋白明显低于对照组;CD47-siRNA转染组食道癌细胞增殖率显著低于对照组(p<0.05);CD47-siRNA转染组细胞增殖相关蛋白PCNA低于对照组(p<0.01);CD47-siRNA转染组食道癌细胞中ROS水平明显高于对照组(p<0.05)。本研究初步认为:CD47-siRNA可降低食道癌细胞中CD47蛋白表达,抑制食道癌细胞的增殖并增加食道癌细胞中ROS水平。     

C-myc-siRNA通过调控ROS诱导子宫内膜癌细胞凋亡

为探讨原癌基因C-myc-siRNA对子宫内膜癌细胞凋亡的影响,本研究利用细胞计数盒(cell counting Kit-8, CCK-8)法分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞活力;蛋白免疫印迹法(Western blotting)分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞凋亡蛋白Bax以及抗凋亡蛋白Bcl-2的变化;流式细胞仪分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)活性氧簇(reactive oxygen species, ROS)的变化;蛋白免疫印迹法(Western blotting)分别检测预处理NAC (ROS抑制剂)后,C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞凋亡相关蛋白Bax以及Bcl-2的变化。结果表明:C-myc-siRNA转染子宫内膜癌细胞后,促凋亡相关蛋白Bax的表达显著高于空载体转染组,且抗凋亡相关蛋白Bcl-2的表达明显低于空载体转染组(p<0.05);C-myc-siRNA转染子宫内膜癌细胞后,细胞ROS水平明显增加(p<0.05);NAC预处理显著减弱C-myc-siRNA对子宫内膜癌细胞凋亡的促进作用(p<0.05)。本研究结论表明,C-myc-siRNA能够通过调控ROS诱导子宫内膜癌细胞凋亡。     

miR-25抑制结肠癌细胞凋亡

为了通过结肠癌细胞实验,探究miR-25对结肠癌细胞凋亡的影响,本研究通过qRT-PCR检测结肠癌细胞中miR-25的表达情况;通过CCK-8法检测miR-25抑制剂转染组细胞活力状态;将miR-25抑制剂转染至结肠癌细胞,通过蛋白免疫印迹法检测结肠癌细胞中Bax和Bcl-2蛋白的表达情况;通过Caspase-3活性检测试剂盒检测结肠癌细胞中Caspase-3表达情况;通过流式细胞仪检测结肠癌细胞中ROS水平;通过Elisa试剂盒检测4-HNE、GSH和MDA含量。结果表明:结肠癌细胞中miR-25表达明显高于正常结肠粘膜组;miR-25抑制剂转染结肠癌细胞后,结肠癌细胞活力明显低于空载体对照组;miR-25抑制剂转染后,结肠癌细胞中Bax蛋白显著高于空载体对照组(p0.05),且Bcl-2蛋白低于空载体对照组(p0.05),Caspase-3活性高于空载体对照组(p0.05);miR-25抑制剂转染后,结肠癌细胞中ROS水平明显高于空载体对照(p0.05);miR-25抑制剂转染后,结肠癌细胞中ROS水平明显高于对照组(p0.05);miR-25抑制剂转染后结肠癌细胞中4-HNE (p0.05)和MDA (p0.05)含量明显增加,GSH (p0.05)含量显著降低。miR-25抑制剂可促进结肠癌细胞凋亡,其机制可能与增加结肠癌细胞中ROS水平导致氧化应激水平增强有关。     

C-myc对子宫内膜细胞增殖和凋亡的影响

为了通过子宫内膜细胞实验检测原癌基因C-myc对子宫内膜癌发生发展的影响,本研究用蛋白免疫印迹法分别检测C-myc转染组和空白对照组中C-myc蛋白的表达情况;细胞计数盒(Cell Counting Kit-8,CCK-8)法分别检测C-myc转染组和空载体转染组的子宫内膜细胞活力;蛋白免疫印迹法分别检测C-myc转染组和空载体转染组的子宫内膜细胞增殖相关蛋白Ki67、凋亡相关蛋白Bax以及Bcl-2的变化;Annexin/PI流式分别检测C-myc转染组和空载体转染组的子宫内膜细胞凋亡率的变化。C-myc转染子宫内膜细胞后,细胞中C-myc蛋白表达明显高于空白对照组;C-myc转染子宫内膜细胞后,细胞活力显著低于空载体转染组(p0.05),Ki67蛋白的表达明显高于空载体转染组(p0.05);C-myc转染子宫内膜细胞后,促凋亡相关蛋白Bax的的表达显著高于空载体转染组(p0.05)以及抗凋亡相关蛋白Bcl-2的表达明显低于空载体转染组(p0.05);C-myc转染子宫内膜细胞后,细胞凋亡率明显高于空载体转染组(p0.05)。本研究表明,C-myc能够促进子宫内膜细胞增殖并诱导细胞凋亡,子宫内膜细胞的增殖过度和凋亡失衡在子宫内膜癌的发生发展中具有重要意义。     

miR-34a在幼鼠海马神经元细胞增殖和凋亡中的作用

目的:探讨miR-34a在幼鼠海马神经元细胞增殖凋亡中的作用。方法:分离幼鼠海马神经元细胞,转染miR-34a抑制剂(miR-34a inhibitor)、抑制剂对照(inhibitor control)、miR-34a模拟物(miR-34a mimics)、模拟物对照(mimics control),RT-PCR检测细胞中miR-34a表达水平。MTT检测转染后细胞增殖情况。流式细胞仪检测细胞凋亡情况。Western blot检测细胞中Cleaved-caspase-3、Bcl-2、Bax的表达水平。结果:转染miR-34a inhibitor可以抑制miR-34a的表达,miR-34a mimics可以促进miR-34a的表达。miR-34a mimics对细胞增殖抑制率明显高于mimics control组(P0.05),miR-34a inhibitor组抑制率明显低于inhibitor control组(P0.05)。miR-34a inhibitor组神经元细胞凋亡率明显低于inhibitor control组(P0.05),miR-34a mimics组神经元细胞凋亡率明显高于mimics control组(P0.01),inhibitor control组和mimics control组神经元细胞凋亡率差异不显著(P0.05)。miR-34a inhibitor组Cleaved-caspase-3、Bax蛋白表达量低于inhibitor control组,差异显著(P0.05);miR-34a inhibitor组Bcl-2蛋白表达量高于inhibitor control组,差异显著(P0.05);miR-34a mimics组Cleaved-caspase-3、Bax蛋白表达量高于mimics control,差异显著(P0.05);miR-34a mimics组Bcl-2蛋白表达量低于mimics control,差异显著(P0.05)。结论:miR-34a抑制海马神经元细胞增殖,促进细胞凋亡,其作用机制可能与调控Cleaved-caspase-3、Bcl-2、Bax表达有关。     

桃叶珊瑚苷对光损伤HaCaT细胞Bax, Bcl-2, Caspase-3表达的影响

目的:探讨桃叶珊瑚苷对紫外线B诱导的HaCaT细胞凋亡相关蛋白Bax、Bcl-2、Caspase-3表达的影响,进一步阐明桃叶珊瑚苷抗光老化的作用机制。方法:将指数生长期HaCaT细胞随机分为正常对照组、模型对照组、10~(-7)、10~(-6)、10~(-5) mol·L~(-1) AU处理组,采用64 m J·cm~(-2)的UVB照射建立细胞光损伤模型,以终浓度10~(-7)、10~(-6)、10~(-5)mol·L~(-1)AU处理光损伤细胞,试剂盒检测ROS含量、Caspase-3活性,流式细胞术检测细胞凋亡率,Western-Blot检测Bcl-2、Bax蛋白的表达量。结果:UV照射的HaCaT细胞ROS含量、细胞凋亡率、Caspase-3活性、Bcl-2、Bax蛋白表达量均升高(P0.01),Bcl-2/Bax值降低(P0.01);不同浓度桃叶珊瑚苷处理后,ROS含量、Caspase-3活性、Bax蛋白表达量降低,Bcl-2蛋白表达量、Bcl-2/Bax值升高,10~(-6)、10~(-5) mol·L~(-1)桃叶珊瑚苷组与模型对照组相比,差异具有统计学意义(P0.05,0.01)。结论:桃叶珊瑚苷通过清除ROS,下调Bax、Caspase-3表达,上调Bcl-2表达,抑制细胞凋亡,保护受损的HaCaT细胞,拮抗光老化。     

转录因子FOXM1在卵巢癌细胞中的表达及其对侵袭转移能力的影响

目的:研究上皮性卵巢癌细胞中FOXM1的表达,探讨FOXM1与MMP9之间的相关性以及与卵巢癌细胞侵袭、转移的关系。方法:采用Real-time RT-PCR、Western blotting技术检测pcDNA3.1-FOXM1和FOXM1-siRNA分别转染卵巢癌细胞株HO-8910(低转移)和HO-8910PM(高转移)前后FOXM1的表达水平,用Transwell方法检测转染该序列后HO-8910和HO-8910PM细胞侵袭能力的改变,并用荧光素酶双报告基因分析技术检测FOXM1对MMP9的调控作用。结果:与对照组和空载组相比,转染了pcDNA3.1-FOXM1的HO-8910细胞FOXM1 mRNA、蛋白表达显著升高,而转染了FOXM1-siRNA的高转移细胞株HO-8910PM FOXM1 mRNA、蛋白表达显著降低(P0.05);相对于空载体组和空白组,pcDNA3.1-FOXM1转染组细胞的侵袭能力明显增强,而FOXM1-siRNA转染细胞的侵袭能力明显降低(P0.05);FOXM1参与对MMP9的转录调控作用(P0.05)。结论:FOXM1可能是一个潜在的治疗靶点,通过下调FOXM1的表达,从而抑制卵巢癌的浸袭和转移。     

lncRNA PVT1沉默对高糖诱导的血管内皮细胞增殖、凋亡及氧化应激的影响*

目的: 探讨抑制lncRNA PVT1对高糖诱导的血管内皮细胞的增殖,凋亡和氧化应激的影响。方法: 体外培养人脐静脉内皮细胞(HUVECs),分为四组:对照组(5.5 mmol/L葡萄糖),高糖组(30 mmol/L葡萄糖),高糖+siNC组(30 mmol/L葡萄糖+siNC,细胞转染阴性对照组),高糖+siPVT1组(30 mmol/L葡萄糖+siPVT1,抑制lncRNA PVT1组)。采用荧光定量PCR的方法检测转染后PVT1的表达水平。MTT检测siPVT1(短片段干扰RNA PVT1)对高糖诱导的HUVECs细胞增殖能力的影响。流式细胞术检测siPVT1对高糖诱导的HUVECs细胞ROS和凋亡水平。Western blot检测HUVECs细胞中凋亡相关蛋白如Bax,Bcl-2和cleaved-caspase-3的表达水平。结果: 与对照组比较,转染siPVT1后,PVT1的表达水平显著降低(P＜0.05)。MTT结果显示,与对照组比较,培养24 h和48 h后高糖组中HUVECs细胞增殖活力均显著降低,与高糖+siNC组(阴性对照组)比较,培养24 h和48 h后,高糖+siPVT1组中的HUVECs细胞增殖活力显著增加(P＜0.05)。流式细胞术检测结果表明,与对照组比较,高糖组HUVECs细胞中ROS和凋亡率均显著增加;和高糖+siNC组比较,高糖+siPVT1组中HUVECs细胞中ROS和凋亡率均有减少(P＜0.05)。Western blot结果表明,与对照组比较,高糖组中cleaved-caspase-3和Bax表达水平均显著上调,Bcl-2的表达水平显著下调(P＜0.05,P＜0.01)。与高糖+siNC组比较,高糖+siPVT1组cleaved-caspase-3和Bax表达水平显著下调,Bcl-2的表达显著上调(P＜0.05,P＜0.01)。结论: 抑制lncRNA PVT1可以显著增加高糖诱导的HUVECs细胞增殖活力,减轻氧化应激,抑制细胞凋亡。     

UBE2T基因对结直肠细胞增殖和凋亡的影响

目的:探讨人类泛素结合酶E2T(Ubiquitin-conjugating enzyme E2T,UBE2T)基因对结肠细胞增殖和凋亡的影响。方法:体外培养人正常结直肠粘膜细胞FHC,采用将UBE2T基因慢病毒质粒转染至FHC细胞48 h后,通过MTT法检测细胞增殖情况,western blotting检测细胞中增殖相关蛋白UBE2T蛋白、Ki67、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达,流式细胞术检测细胞凋亡率。结果:与转染空质粒的FHC细胞相比,UBE2T基因慢病毒质粒转染FHC细胞48 h后,细胞增殖能力显著上调(P0.05),UBE2T蛋白明显增加,Ki67的表达明显增加(P0.05),细胞凋亡率显著降低(P0.05),且Bax的表达明显下调而Bcl-2的表达上调(P0.05)。结论:UBE2T基因能够促进正常结肠粘膜细胞的增殖,并抑制其凋亡。     

氧化槐果碱通过调控lncRNA DUXAP8/miR-519b-3p轴影响宫颈癌细胞增殖、凋亡、迁移及侵袭的实验研究

为了探讨氧化槐果碱(OSC)对宫颈癌HeLa细胞增殖、凋亡、迁移和侵袭的影响及作用机制,本研究将HeLa细胞分为对照组(Con组)、OSC组、转染DUXAP8小干扰RNA(si-DUXAP8)组、转染小干扰RNA阴性对照(si-NC)组、OSC+DUXAP8过表达载体(pcDNA-DUXAP8)组和OSC+空载体(pcDNA)组,然后用MTT法检测细胞增殖,Transwell检测细胞迁移和侵袭,流式细胞术检测细胞凋亡,蛋白印迹法检测细胞中CyclinD1、p21、Bcl-2、Bax、MMP-2和MMP-9蛋白水平,RT-qPCR检测细胞中DUXAP8和miR-519b-3p水平;双荧光素酶报告基因实验验证DUXAP8和miR-519b-3p调控关系。结果显示,与Con组比较,OSC组HeLa细胞抑制率、凋亡率、Bax和p21蛋白水平、miR-519b-3p水平升高(P0.05),细胞迁移数、侵袭数、CyclinD1、Bcl-2、MMP-2和MMP-9的蛋白水平和DUXAP8水平降低(P0.05),且呈剂量依赖性(P0.05);与si-NC组比较,si-DUXAP8组HeLa细胞抑制率、凋亡率和Bax、p21蛋白水平升高(P0.05),细胞迁移和侵袭数及CyclinD1、Bcl-2、MMP-2、MMP-9的蛋白水平降低(P0.05);DUXAP8在HeLa细胞中靶向负调控miR-519b-3p表达;与OSC+pcDNA组比较,OSC+pcDNA-DUXAP8组HeLa细胞抑制率、凋亡率和Bax、p21的蛋白水平降低(P0.05),细胞迁移和侵袭数及CyclinD1、Bcl-2、MMP-2、MMP-9的蛋白水平升高(P0.05)。这提示氧化槐果可能通过调DUXAP8/miR-519b-3p轴抑制宫颈癌HeLa细胞增殖、迁移和侵袭,并诱导细胞凋亡。     

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).