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1.
目的:探讨miR-194对乳腺癌细胞增殖、侵袭迁移和Wnt/β-catenin信号通路的影响及作用机制。方法:将MCF-7细胞分为空白对照组(Blank组)、阴性对照组(NC组)和miR-194组。miR-194组转染miR-125b mimic,NC组转染NC,Blank组不转染。RT-PCR测定细胞中miR-194水平,MTT法测定细胞增殖活性,Transwell小室测定细胞侵袭能力,划痕实验测定细胞迁移能力,Western blot测定细胞中糖原合成酶激酶3β(GSK-3β)、磷酸化GSK-3β(p-GSK-3β)、β-连环蛋白(β-catenin)、细胞周期蛋白D1(cyclin-D1)、C-Myc、基质金属蛋白酶7(MMP-7)蛋白水平。结果:乳腺癌MCF-7细胞中miR-194水平低于正常乳腺Hs578Bst细胞(P0.05)。与Blank组和NC组相比,miR-194组miR-194水平升高(P0.05),OD值、侵袭细胞数减少、迁移距离缩短(P0.05),p-GSK-3β、β-catenin、cyclin-D1、C-Myc、MMP-7蛋白水平降低(P0.05),Blank组和NC组各指标差异无统计学意义(P0.05)。结论:乳腺癌细胞中miR-194水平降低,过表达miR-194可通过抑制Wnt/β-catenin信号通路抑制乳腺癌细胞的增殖和侵袭迁移。  相似文献   

2.
目的:探讨microRNA-373在乳腺癌组织中表达与侵袭转移的相关性。方法:建立乳腺癌细胞MCF-7裸鼠移植瘤模型,采用qRT-PCR法检测癌组织和癌旁组织中miRNA-373的表达。分别转染空白试剂、miR-373阴性对照物、miR-373拟似物、miR-373阻遏物至MCF-7细胞,采用荧光实时定量PCR法检测细胞内miRNA-373含量,采用Transwell小室法检测MCF-7细胞侵袭能力。结果:qRT-PCR检测显示,乳腺癌组织miRNA-373表达显著低于癌旁组织,差异有统计学意义(P 0. 05)。qRT-PCR检测显示,转染后BC组和NC组miRNA-373表达差异无统计学意义(P0. 05),MT组miRNA-373表达显著高于BC组、NC组和RT组(P0. 05),RT组miRNA-373表达显著低于BC组、NC组和MT组(P0. 05)。MT组穿膜细胞数显著低于BC组、NC组和RT组,RT组穿膜细胞数显著高于BC组、NC组和MT组,差异均有统计学意义(P0. 05); BC组和NC组穿膜细胞数相比较差异无统计学意义(P 0. 05)。结论:乳腺癌MCF-7细胞的侵袭和迁移能力受miR-373调控,增强miRNA-373可降低MCF-7细胞侵袭和迁移能力,为乳腺癌治疗的新基因靶点药物的开发提供了思路。  相似文献   

3.
目的:探讨miR-610 对卵巢癌细胞侵袭和迁移能力的作用及机制。方法:通过RT-PCR 检测卵巢癌组织及细 胞、正常组织及细胞中miR-610 与鼠双微体基因2(MDM2)mRNA的表达;通过Lipofectamine 2000 将miR-NC、 miR-610 mimic、miR-6101 inhibitor、pc-MDM2质粒分别或联合转染进入HO8910细胞;划痕实验检测细胞迁移 能力,Transwell 法检测细胞侵袭能力,免疫印迹检测E- 钙黏着蛋白(E-cadherin)、N- 钙黏着蛋白(N-cadherin) 和MDM2蛋白表达水平,双荧光素酶报告基因检测miR-610 与MDM2靶向关系。 结果:与正常卵巢组织和上皮 细胞系HOSEpiC相比,人宫颈癌组织和细胞系中miR-610 表达明显下调,选择下调效果较明显的HO8910细胞系 进行后续实验。与Control 组相比,miR-610 mimic 组卵巢癌HO8910细胞划痕闭合率明显降低、侵袭细胞数目明 显减少、E-cadherin 明显上调、N-cadherin 表达明显下调,miR-610 inhibitor 组卵巢癌HO8910细胞划痕闭合率明显 升高、侵袭细胞数目明显增多、E-cadherin 明显下调、N-cadherin 表达明显上调。MiR-610 与MDM2 3'UTR 区存在 结合位点,且miR-610 靶向下调MDM2 表达。共转染pc-MDM2逆转了miR-610 mimic 对卵巢癌HO8910细胞迁移 和侵袭能力抑制作用。 结论:MiR-610 通过靶向下调MDM2抑制卵巢癌HO8910细胞迁移和侵袭能力。  相似文献   

4.
目的:探讨微小RNA-137(miR-137)对乳腺癌细胞侵袭、迁移及凋亡的影响和分子机制。方法:在乳腺癌MDA-MB-231细胞中转染miR-137模拟物(miR-137 mimics),RT-qPCR检测miR-137表达量,流式细胞术检测细胞凋亡,Transwell小室检测侵袭和迁移,Western blot检测基质金属蛋白酶9(MMP-9)、cleaved caspase-3(C-caspase-3)和Bax的蛋白水平。生物信息学软件预测TWIST1可能是miR-137的靶基因后用萤光素酶报告基因鉴定。Western blot检测miR-137 mimics对TWIST1蛋白表达影响。在乳腺癌细胞中共转染TWIST1过表达载体和miR-137 mimics,测定细胞凋亡、侵袭、迁移及MMP-9、C-caspase-3、Bax蛋白水平的变化。结果:转染miR-137 mimics后的乳腺癌细胞中miR-137水平升高,细胞凋亡率升高,细胞侵袭和迁移能力降低,C-caspase-3和Bax的蛋白水平增高,MMP-9蛋白表达减少(P0.05)。萤光素酶报告载体鉴定显示miR-137靶向调控TWIST1,并且miR-137 mimics能够抑制乳腺癌细胞中TWIST1表达。与共转染阴性对照载体和miR-137 mimics的细胞比较,TWIST1过表达载体和miR-137 mimics共转染后的乳腺癌细胞TWIST1和MMP-9蛋白表达水平升高,C-caspase-3和Bax蛋白水平减少,细胞凋亡率降低,细胞侵袭和迁移能力增强(P0.05)。结论:miR-137靶向TWIST1抑制乳腺癌细胞侵袭迁移并诱导细胞凋亡。  相似文献   

5.
目的 探讨miR-138对三阴型乳腺癌(triple-negative breast cancer, TNBC)MDA-MB-231细胞侵袭、转移的影响及分子机制。方法 将MDA-MB-231细胞转染miR-138 mimics及其阴性对照,Transwell实验检测细胞迁移和侵袭能力改变。生物信息学预测miR-138的潜在靶基因,采用双荧光素酶报告实验和Western blot法分析miR-138与FAK的靶向调控关系。回复实验使用Transwell及Western blot实验验证miR-138通过FAK抑制MDA-MB-231细胞侵袭、转移。结果 Transwell实验结果表明:过表达miR-138后,MDA-MB-231细胞的迁移和侵袭能力均显著降低(P<0.05)。生物信息学预测FAK为miR-138的潜在靶基因,双荧光素酶报告实验结果表明:FAK 3′UTR区域存在miR-138的互补结合位点。Western blot结果发现过表达miR-138后FAK蛋白表达降低。回复实验Transwell及Western blot结果表明:上调FAK可以部分恢复miR-138过表...  相似文献   

6.
目的:以人绒毛膜滋养层细胞系JEG-3 细胞为研究对象,结合该细胞侵袭和迁移能力的变化情况,研究转染miR-155 mimics 和miR-155 inhibitor 之后CXCR4 的表达变化及其对下游PI3K/ AKT 信号通路的影响作用,从而探讨miR-155参与子痫前期发生发展的分子机制。方法:设计miR-155mimics 和miR-155 inhibitor,对JEG-3 进行转染,通过Transwell 侵袭实验、划痕实验,观察转染后细胞的侵袭和迁移能力的变化;利用Real-time PCR 检测CXCR4 mRNA 的表达;利用Western blot检测CXCR4 及下游p鄄AKT 蛋白的表达水平。结果:Real-time PCR 结果显示,miR-155 mimics 转染组CXCR4 mRNA 相对表达量(0.589依0.096)明显低于空白对照组(1.503依0.090)和阴性对照组(1.146依0.153),差异有统计学意义(P<0.05);miR-155 inhibitor 转染组CXCR4 mRNA 相对表达量(1.739依0.083)与两组对照组相比差异也有统计学意义(P<0.05)。Western blot 结果显示miR-155 mimics 转染组CXCR4 蛋白和下游p-AKT 蛋白表达水平均明显降低,而miR-155 inhibitor 转染组CXCR4 和p-AKT 蛋白水平则升高,与空白对照组和阴性对照组相比差异均有统计学意义(P<0.05)。Transwell 侵袭实验结果显示,与空白对照组(63郾46依2郾37)和阴性对照组(49.29依5.81)侵袭细胞数相比,miR-155 mimics 转染组侵袭细胞数(22.89依9.42)明显减少,差异有统计学意义(P<0.05);同时,miR-155 inhibitor 转染组侵袭细胞数(81.50依11.25)明显升高,差异有统计学意义(P<0.05)。划痕实验结果显示,转染miR-155 mimics 后,JEG-3 细胞相对迁移距离(0.159依0.058)低于空白对照组(1.080依0.045)和阴性对照组(0.823依0.201),差异有统计学意义(P<0.05);而miR-155 inhibitor 转染组,JEG-3 细胞相对迁移距离(1.640依0.078)明显增加,差异有统计学意义(P<0.05)。结论:miR-155 可能通过抑制CXCR4 的表达进而抑制其下游PI3K/AKT 信号通路的活化,从而影响滋养细胞的侵袭及迁移能力,最终导致子痫前期的发生发展。  相似文献   

7.
目的 探讨微小RNA-425-5p(miR-425-5p)靶向FOXJ3对乳腺癌细胞的作用.方法 用Lipofiectamine2000分别将miR-425-5p inhibitor及NC转染至MDA-MB-231乳腺癌细胞作为miR-425-5p inhibitor组和NC组,不做任何处理的为对照组.通过RT-qPCR测乳腺上皮细胞MCF10A及miR-425-5p inhibitor组、NC组、对照组中miR-425-5p及FOXJ3mRNA表达水平.通过MTT法测miR-425-5p inhibitor组、NC组、对照组细胞增殖情况;通过流式细胞仪测3组细胞凋亡情况;Transwell小室实验测细胞侵袭情况;Western-blot测细胞中FOXJ3、CyclinD1、Bcl-2、MMP-2蛋白表达情况;双荧光素酶实验验证miR-425-5p与FOXJ3的靶向关系.结果 与乳腺上皮细胞MCF10A比,乳腺癌细胞MDA-MB-231中miR-425-5p表达水平显著升高(P<0.05),FOXJ3mRNA表达水平显著下降(P<0.05);与对照组和NC组比,miR-425-5p inhibitor组细胞增殖、迁移、侵袭能力受到抑制,细胞凋亡率升高(P<0.05);双荧光素酶实验结果显示抑制miR-425-5p表达显著增加野生型FOXJ33'UTR萤光素酶活性,对突变型FOXJ33'UTR的萤光素酶活性无影响.结论 下调miR-425-5p可能通过靶向FOXJ3抑制乳腺癌细胞增殖、迁移、侵袭,并促进细胞凋亡.  相似文献   

8.
目的研究七氟烷对人乳腺癌细胞(MCF-7)增殖、迁移和侵袭的影响及其机制。方法用Western blot检测MCF-7细胞中高迁徙率族蛋白1 HMGB1的表达; miR-34a组(转染miR-34a mimics)、miR-con组(转染miR-con)、七氟烷处理+anti-miR-con组(转染anti-miR-con)及七氟烷处理+anti-miR-34a组(转染anti-miR-34a),均用脂质体法转染至MCF-7细胞; RT-qPCR检测细胞miR-34a表达; MTT法检测细胞增殖; Transwell小室法检测细胞迁移和侵袭;双荧光素酶报告基因实验检测细胞荧光活性。结果与对照组相比,1. 7%的七氟烷处理的细胞中miR-34a表达显著升高(P0. 05),HMGB1表达显著降低(P0. 05),且细胞增殖、迁移和侵袭均显著下调(P0. 05);过表达miR-34a可抑制MCF-7细胞增殖、迁移和侵袭,敲减miR-34a可降低七氟烷对MCF-7细胞增殖、迁移和侵袭的抑制作用。miR-34a可明显降低野生型HMGB1的细胞荧光活性,且负向调控HMGB1的表达。结论七氟烷可抑制人乳腺癌细胞增殖、迁移和侵袭。  相似文献   

9.
目的研究miR-451a在膀胱癌中的表达及对膀胱癌发生发展的影响。方法通过实时定量PCR(real-time PCR)检测miR-451a在膀胱癌细胞系及组织标本中的表达。采用real-time PCR和Western blot检测MEF2D在膀胱癌组织及转染miR-451a mimics和inhibitor的T24细胞中的表达。双荧光素酶基因报告实验分析MEF2D是否为miR-451a直接的靶基因。通过MTT,Transwell迁移和侵袭实验检测T24细胞在转入miR-451a mimics和inhibitor后细胞增殖、迁移和侵袭能力的变化。结果 miR-451a在膀胱癌细胞及组织中的表达水平下调。miR-451a抑制了T24细胞的增殖、迁移和侵袭。双荧光素酶基因报告证实MEF2D为miR-451a的直接靶基因。miR-451a能够降低T24细胞MEF2DmRNA和蛋白表达水平。结论 miR-451a在膀胱癌中表达下调,通过靶向调节MDF2D起到了抑癌的作用,可能成为膀胱癌诊疗的潜在靶点。  相似文献   

10.
目的 探讨miR-27a对结肠癌SW480细胞增殖、凋亡、侵袭及迁移的影响及可能机制。方法 qRT-PCR方法检测人结肠癌细胞株SW480与人正常结肠黏膜上皮细胞NCM460中miR-27a的表达水平。体外培养SW480细胞,将SW480细胞随机分为空白对照组(不进行任何处理)、miR-27a抑制剂组(转染miR-27a inhibitor)与阴性对照组(转染miR-27a inhibitor NC),采用脂质体法进行转染,实时荧光定量PCR方法验证转染效率。采用四甲基偶氮唑蓝(MTT)法检测SW480细胞增殖;Transwell实验检测SW480细胞侵袭与迁移;流式细胞术检测SW480细胞凋亡;Western blot法检测转染后叉头框蛋白O1(FOXO1)、组织金属蛋白酶抑制剂2(TIMP-2)、泛素连接酶FBW7的表达。结果 miR-27a在SW480细胞中的表达水平显著高于NCM460细胞(P<0.01)。转染miR-27a抑制剂后,SW480细胞中miR-27a的表达水平明显降低,转染成功。抑制miR-27a表达后,SW480细胞的增殖、迁移与侵袭能力显著降低,凋亡率显...  相似文献   

11.
Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc2 (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.  相似文献   

12.
Renal dysplasia and asplenia in two sibs   总被引:2,自引:0,他引:2  
A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.  相似文献   

13.
About 1900, modern food selection and processing caused widespread epidemics of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a panhypovitaminosis B, i.e., a mixture of substrate beriberi and substrate pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse endemic disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for primates. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.  相似文献   

14.
15.
Newton H 《Medical history》2011,55(2):153-182
Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.  相似文献   

16.
Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.  相似文献   

17.
HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.  相似文献   

18.
The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.  相似文献   

19.
Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.  相似文献   

20.
Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.  相似文献   

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