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1.
目的:讨论姜黄素联合顺铂对卵巢癌耐药细胞增殖、迁移及凋亡能力的作用,并分析其与PI3K/Akt通路的关系。方法:首先用不同浓度的姜黄素及顺铂单独处理细胞株,根据各浓度细胞的存活率确定35µmol/L姜黄素、4.0 mg/L顺铂疗效最好,用于后续实验。将卵巢癌耐药细胞分为空白对照组、姜黄素单独处理组(35µmol/L)、顺铂单独处理组(4.0 mg/L)、姜黄素+顺铂联合处理组(35µmol/L+4.0 mg/L),采用MTT法、Transwell小室、流式细胞术检测四组细胞的生长抑制率、迁移细胞数及细胞凋亡率;Western blot检测四组细胞中p-PI3K及p-Akt蛋白表达水平;PCR检测四组细胞中PI3K、Akt、Ki67、Bcl-2、Bax mRNA表达水平。结果:与空白对照组比较,其余三组细胞生长抑制率及细胞凋亡率均显著升高,迁移细胞数显著减少(P<0.05),且姜黄素+顺铂联合组细胞生长抑制率及细胞凋亡率显著高于姜黄素组、顺铂组,迁移细胞数则低于姜黄素组、顺铂组(P<0.05);与空白对照组比较,姜黄素处理组及姜黄素+顺铂联合组细胞中p-PI3K、p-Akt蛋白及PI3K、Akt、Ki67、Bcl-2 mRNA表达水平均显著降低、Bax mRNA显著升高(P<0.05),且联合组细胞表达量明显低于姜黄素处理组(P<0.05),而顺铂单独处理组细胞中PI3K、p-Akt蛋白及PI3K、Akt、Ki67、Bcl-2、Bax mRNA表达水平无明显变化(P>0.05)。结论:姜黄素联合顺铂能够提高卵巢癌耐药细胞的敏感性,抑制细胞的增殖及迁移,同时诱导细胞凋亡,这一过程可能通过下调PI3K/Akt通路的表达实现。  相似文献   

2.
目的探讨HIPK2对顺铂诱导的人肾小管上皮细胞(HKC)凋亡的影响。方法构建顺铂诱导的HKC细胞凋亡模型,实时荧光定量PCR和Western blot检测HIPK2的表达;设计合成2条HIPK2 siRNA干扰片段,通过脂质体转染HKC细胞,建立HIPK2干扰的细胞株;实时荧光定量PCR和Western blot分别检测HIPK2 mRNA和蛋白的表达;再用顺铂处理细胞,Annexin V/PI检测细胞凋亡;Western blot检测Bax表达的影响。结果顺铂呈剂量依赖性诱导的HKC细胞凋亡过程中,HIPK2 mRNA和蛋白表达水平均明显下调(P0.05);转染siRNA后可显著降低HIPK2在HKC细胞内mRNA和蛋白的表达(P0.05),且HIPK2表达降低后会促进顺铂诱导的HKC细胞凋亡。结论HIPK2可抑制顺铂诱导的HKC细胞凋亡。  相似文献   

3.
目的研究虫草素与顺铂联合用药对食管癌细胞Eca109凋亡的影响及其作用机制。方法将食管癌细胞Eca109分为对照组、不同浓度虫草素处理组、不同浓度顺铂处理组和虫草素(70μg/m L)与顺铂(0.8μg/m L)联合处理组。MTS法检测Eca109细胞增殖;Hoechst 33258染色法及流式细胞术检测Eca109细胞凋亡;Western blot法检测细胞核内NF-κB P65及凋亡相关蛋白Bcl-2和Bax表达水平;ELISA法检测细胞核内NF-κB P65与DNA的结合活性。结果虫草素与顺铂联合应用使顺铂对Eca109细胞的抑制率由29.30%增加至70.41%(P0.05),增强了顺铂对Eca109的敏感性;与顺铂单独用药相比,联合用药能够显著增加顺铂诱导的细胞凋亡(P0.05);与对照组相比顺铂能够增加细胞核内NF-κB P65的活性,而虫草素却能抑制NF-κB P65的活性,联合用药后NF-κB P65的活性下调;与虫草素和顺铂单独用药相比,联合用药组能够使抗凋亡蛋白Bcl-2的表达显著降低(P0.05),而促凋亡蛋白Bax表达则显著增高(P0.05)。结论虫草素可能通过抑制NF-κB途径,调节下游信号分子Bcl-2和Bax的表达,增强顺铂对Eca109的凋亡诱导效应。  相似文献   

4.
目的探讨丙酮酸激酶2(PKM2)在脂多糖(LPS)诱导人肾小管上皮细胞(HK-2)凋亡中的作用及分子机制。方法采用不同质量浓度的脂多糖作用于HK-2细胞,利用CCK-8实验检测细胞存活率;利用Real-time PCR、Western blot检测HK-2细胞中PKM2的mRNA及蛋白表达水平;利用Western blot检测Bcl-2家族关键蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)以及细胞色素C(Cytochrome C)的表达;利用Annexin-PE/7AAD双染法分析细胞凋亡水平;利用流式细胞术PI染色法检测细胞周期,并结合PKM2抑制剂,深入探讨PKM2在其中的作用及机制。结果脂多糖可诱导HK-2细胞损伤,使其活性降低、周期阻滞、凋亡增加,且可检测到Bcl-2家族关键蛋白的表达改变:Bcl-2降低、Bax增高,同时伴Cytochrome C增加。脂多糖可诱导PKM2表达增加,而使用PKM2抑制剂后,进一步增加了HK-2细胞凋亡以及细胞G1-S期阻滞,伴Bcl-2显著降低,Bax、Cytochrome C显著增加。结论脂多糖诱导HK-2细胞活力下降,周期阻滞,凋亡增加,PKM2可以通过靶向Bcl-2家族参与脂多糖诱导的HK-2细胞凋亡。  相似文献   

5.
目的:观察沉默miR-21对子宫内膜癌顺铂耐药细胞株Ishikawa/DDP的影响.方法:以Lipofectamine 2000介导miR-21抑制剂转染Ishikawa/DDP细胞株,同时设置阴性组和耐药组.采用反转录PCR检测miR-21、多药耐药基因MDR1、促凋亡基因Bax和抗凋亡基因Bcl-2的表达.采用蛋白印迹法检测多药耐药蛋白P-gp、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达.采用噻唑蓝比色法检测细胞对顺铂的敏感性.采用流式细胞术检测细胞凋亡情况.结果:与耐药组和阴性组比较,抑制剂组miR-21,MDR1和Bcl-2 mRNA表达显著下调(P<0.0l),而Bax mRAN表达显著上调(P<0.00l);抑制剂组P-gp和Bcl-2蛋白表达显著低下调(p<0.05),而Bax蛋白表达显著上调(P<0.001).与耐药组和阴性组比较,顺铂对抑制剂组的IC50值显著(P<0.001);顺铂对抑制剂组细胞的诱导凋亡率显著增加(P<0.00l).结论:沉默miR-21可显著提高Ishikawa/DDP细胞株对顺铂的敏感性,并促进细胞凋亡,其具体机制可能与下调MDR1,P-gp和Bcl-2表达,以及上调Bax表达有关.  相似文献   

6.
目的:观察中药单体姜黄素对ActD/TNF-α协同诱导PC12细胞凋亡的影响,并探讨其机制。方法:采用MTT法确定实验药物的最佳浓度;Hoechst33258荧光染色法观察PC12细胞的凋亡;JC-1荧光分子探针检测线粒体膜电位;Real Time PCR检测凋亡基因Bcl-2/Bax的表达。结果:ActD/TNF-α协同作用可导致PC12细胞的活力降低(P<0.05);出现核固缩、核碎裂现象的细胞增多,细胞凋亡率增高(P<0.05);细胞线粒体膜电位下降;细胞内抗凋亡基因Bcl-2的表达降低(P<0.05)。经姜黄素(5μmol/L)处理后,PC12细胞的活力增强(P<0.05);细胞核固缩、核碎裂现象减少,细胞凋亡率下降(P<0.05);细胞线粒体膜电位上升;细胞内抗凋亡基因Bcl-2的表达增强(P<0.05)。结论:姜黄素可拮抗ActD/TNF-α引起的PC12细胞凋亡,可能与升高线粒体膜电位,促进抗凋亡基因Bcl-2的表达有关。  相似文献   

7.
目的:探讨姜黄素对缺氧条件下大鼠星型胶质细胞活力、细胞凋亡率及JAK2/STAT3信号通路的影响。方法:从大鼠大脑皮层分离培养星形胶质细胞,姜黄素干预缺氧条件星形胶质细胞,AG490作为JAK2/STAT3信号通路抑制剂,细胞缺氧处理24 h后,通过MTT法检测各组细胞活力;流式细胞术检测细胞凋亡率;Western blot检测凋亡蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、JAK2/STAT3信号通路磷酸化的蛋白酪氨酸激酶2(p-JAK2)及磷酸化的信号转导与转录因子3(p-STAT3)的蛋白表达。结果:与对照组比较,缺氧组细胞活力显著升高,凋亡率显著升高,Bcl-2蛋白表达显著降低,Bax、p-JAK2和p-STAT3的蛋白表达显著升高(P<0.05);与缺氧组比较,缺氧+姜黄素组胞活力显著降低,凋亡率显著降低,Bcl-2蛋白表达显著升高,Bax、p-JAK2和p-STAT3的蛋白表达显著降低(P<0.05);与缺氧+姜黄素组比较,缺氧+姜黄素组+AG490组细胞活力显著降低,细胞凋亡率显著降低,Bcl-2蛋白表达显著升高,Bax、p-JAK2和p-STAT3的蛋白表达显著降低(P<0.05)。结论:姜黄素可降低缺氧下大鼠星型胶质细胞活力,抑制细胞凋亡,其机制与JAK2/STAT3信号通路有关。  相似文献   

8.
目的探讨脑源性神经营养因子反义RNA(brain-derived neurotrophic factor-antisense, BDNF-AS)对高糖诱导的肾小管上皮细胞损伤的影响和可能机制。方法体外培养肾小管上皮细胞HK-2, 分别转染BDNF-AS小干扰RNA、miR-145-5p模拟物或共转染BDNF-AS小干扰RNA和miR-145-5p抑制剂, 之后采用30 mmol/L葡萄糖干预转染后的细胞24 h, 用RT-qPCR法检测细胞中BDNF-AS和miR-145-5p的表达, 用CCK-8法检测细胞增殖, 流式细胞术检测细胞凋亡, Western印迹法检测细胞中Bcl-2和Bax蛋白的表达, 酶联免疫吸附法检测细胞培养上清中IL-1β和IL-6的水平。用双荧光素酶报告基因实验验证BDNF-AS和miR-145-5p的调控关系。结果高糖处理促进了HK-2细胞中BDNF-AS的表达(P<0.05), 而抑制了miR-145-5p的表达(P<0.05)。干扰BDNF-AS或过表达miR-145-5p降低了高糖诱导的HK-2细胞抑制率、凋亡率及Bax蛋白、IL-1β和IL-6的表达(P<0.05), 促进了Bcl-2蛋白的表达(P<0.05)。干扰miR-145-5p逆转了干扰BDNF-AS对高糖诱导的HK-2细胞增殖、凋亡及IL-1β和IL-6表达的影响。BDNF-AS可靶向结合并负调控miR-145-5p。结论干扰BDNF-AS可能通过靶向负调控miR-145-5p促进高糖诱导的肾小管上皮细胞增殖, 并抑制细胞凋亡及炎症因子表达。  相似文献   

9.
目的探讨脑源性神经营养因子反义RNA(brain-derived neurotrophic factor-antisense, BDNF-AS)对高糖诱导的肾小管上皮细胞损伤的影响和可能机制。方法体外培养肾小管上皮细胞HK-2, 分别转染BDNF-AS小干扰RNA、miR-145-5p模拟物或共转染BDNF-AS小干扰RNA和miR-145-5p抑制剂, 之后采用30 mmol/L葡萄糖干预转染后的细胞24 h, 用RT-qPCR法检测细胞中BDNF-AS和miR-145-5p的表达, 用CCK-8法检测细胞增殖, 流式细胞术检测细胞凋亡, Western印迹法检测细胞中Bcl-2和Bax蛋白的表达, 酶联免疫吸附法检测细胞培养上清中IL-1β和IL-6的水平。用双荧光素酶报告基因实验验证BDNF-AS和miR-145-5p的调控关系。结果高糖处理促进了HK-2细胞中BDNF-AS的表达(P<0.05), 而抑制了miR-145-5p的表达(P<0.05)。干扰BDNF-AS或过表达miR-145-5p降低了高糖诱导的HK-2细胞抑制率、凋亡率及Bax蛋白、IL-1β和IL-6的表达(P<0.05), 促进了Bcl-2蛋白的表达(P<0.05)。干扰miR-145-5p逆转了干扰BDNF-AS对高糖诱导的HK-2细胞增殖、凋亡及IL-1β和IL-6表达的影响。BDNF-AS可靶向结合并负调控miR-145-5p。结论干扰BDNF-AS可能通过靶向负调控miR-145-5p促进高糖诱导的肾小管上皮细胞增殖, 并抑制细胞凋亡及炎症因子表达。  相似文献   

10.
 目的 研究赖氨大黄酸(RHL)、顺铂和两者联合对人肺癌A549细胞增殖和凋亡的影响及其作用机制,为RHL联合顺铂抗肺癌的临床实验研究提供理论依据。方法 肺癌细胞株A549随机分为对照组、顺铂组、RHL组和顺铂联合RHL组,用MTT法和流式细胞术分别检测细胞48 h增殖和凋亡。Western blot 法检测细胞48h后凋亡相关蛋白和MEK/ERK蛋白的表达。结果 细胞培养48 h后,顺铂组和RHL组的细胞增殖受到一定的抑制并有细胞凋亡发生,但两药联合后的增殖抑制作用和凋亡诱导作用显著高于单用顺铂和RHL组(P<0.05)。顺铂和RHL均能上调caspase-3、caspase-7和poly ADP-ribose polymerase (PARP)的切割片断蛋白表达,但两药联合后caspase-3、caspase-7和PARP的切割片断蛋白表达进一步增高(P<0.05),并显著下调Bcl-2蛋白的表达水平,上调Bax蛋白的表达水平,同时RHL还能降低顺铂上调的ERK蛋白磷酸化。结论 RHL能够通过抑制顺铂对ERK的激活、上调caspase-3、caspase-7和PARP的切割片断蛋白表达,增强顺铂对肺癌细胞增殖和凋亡诱导作用。  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   

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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.
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   

18.
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   

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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