MG132抑制TGF-β1诱导的人肺成纤维细胞活化

目的探讨泛素-蛋白酶体抑制剂MG132对转化生长因子β1(TGF-β1)诱导人肺成纤维细胞活化的影响及机制。方法体外培养人胚肺成纤维细胞(MRC-5),随机分为对照组、TGF-β1组(10μg/L)和MG132(0.5μmol/L)处理组。Western blot法检测细胞α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原α1(COL1A1)的表达;RT-PCR和Western blot法分别检测细胞Smad7、核转录共抑制因子SnoN、TGF-βI型受体(TβRI)、Smad2和Smad3 mRNA及蛋白的表达。结果与对照组比较,TGF-β1促进了α-SMA和COL1A1蛋白表达(P0.05),MG132抑制了TGF-β1的上述作用(P0.05)。TGF-β1组Smad7和SnoN mRNA表达较对照组增加(P0.05)。TGF-β1组Smad7和SnoN蛋白表达较对照组减少(P0.05),而MG132组Smad7和SnoN蛋白水平较TGF-β1组增加(P0.05)。结论 MG132可能通过阻止Smad7和SnoN蛋白泛素化降解,抑制TGF-β1诱导人肺成纤维细胞活化。     

Smad4的表达与胰腺癌淋巴管生成之间的关系

目的观察血管内皮生长因子(VEGF)-C和Smad 4在胰腺癌组织内的表达情况,分析VEGF-C和Smad 4的表达与胰腺癌淋巴管生成及淋巴结转移之间的关系。方法取胰腺癌病例52例,其中,无淋巴结转移组12例,淋巴结转移组40例。应用免疫组化和Western blot技术检测VEGF-C和Smad4在胰腺癌组织内的表达情况。以D2-40作为淋巴管内皮标记物,观察胰腺癌组织内淋巴管生成情况。结果 VEGF-C主要表达于胰腺癌细胞胞质内,在淋巴结转移组的表达水平明显高于无淋巴结转移组。Smad4表达于胰腺癌细胞胞质和胞核内,在无淋巴结转移组的表达水平明显高于淋巴结转移组,Smad4表达阳性组淋巴管数密度明显低于Smad4表达阴性组(=0.02)。Smad4的表达与VEGF-C的表达呈显著的负相关性。结论 VEGF-C在胰腺癌淋巴管的发生及淋巴结转移中发挥重要作用,Smad4可能通过调节VEGF-C蛋白的表达抑制胰腺癌淋巴管生成和淋巴结转移。     

EGCG联合盐酸吉西他滨通过下调PAR-2诱导胰腺癌细胞凋亡

目的评价表没食子儿茶素没食子酸酯(EGCG)联合盐酸吉西他滨对胰腺癌PANC-1细胞凋亡及对蛋白酶活化受体2(PAR-2)表达的影响。方法培养PANC-1细胞,分为不添加任何药物的对照组、60μg/mL EGCG处理组、20μg/mL吉西他滨处理组、60μg/mL EGCG联合20μg/mL吉西他滨处理组。Annexin V-FITC/PI联合染色结合流式细胞术检测各组PANC-1细胞凋亡情况,并采用实时荧光定量PCR(qRT-PCR)检测各组PANC-1细胞PAR-2 mRNA的表达。Western blot法检测各组PANC-1细胞PAR-2蛋白的表达。结果联合处理组对PANC-1细胞凋亡的诱导作用最强,依次为EGCG处理组、吉西他滨处理组。各组与对照组相比,差异均有统计学意义(P0.01)。qRT-PCR和Western blot法结果表明联合处理组的PAR-2 mRNA和蛋白的相对表达量低于对照组(P0.01或P0.05)。结论 EGCG可以增强盐酸吉西他滨对胰腺癌PANC-1细胞凋亡的诱导作用,该作用与下调细胞PAR-2表达有关。     

c-SKI对冠脉内皮细胞增殖及内皮-间充质转化的影响

目的:探讨核蛋白c-SKI对冠状动脉内皮细胞增殖能力及内皮-间充质转化的影响。方法:使用不同浓度的转化生长因子β1(TGF-β1)作用人冠脉内皮细胞不同时点,Western blot法检测各组细胞中c-SKI、间充质细胞标志物波形蛋白和α-平滑肌肌动蛋白(α-SMA)及内皮细胞标志物E-钙黏蛋白的表达。使用携带c-ski基因的慢病毒转染冠脉内皮细胞后采用RT-qPCR验证转染效率。将细胞分为4组:对照组、TGF-β1(5μg/L)组、c-ski基因感染+TGF-β1组及对照病毒感染+TGF-β1组。过表达c-SKI后,MTT实验和集落形成实验检测冠脉内皮细胞的增殖能力,Western blot检测波形蛋白、α-SMA、E-钙黏蛋白、Smad2、Smad3、p-Smad2和p-Smad3蛋白的水平。结果:TGF-β1处理冠脉内皮细胞后,c-SKI在冠脉内皮细胞中的表达呈剂量和时间依赖性下降(P0.01)。MTT及集落形成实验结果显示,过表达c-SKI可显著抑制冠脉内皮细胞增殖(P0.01)。Western blot检测结果显示,与病毒对照组相比,过表达c-SKI可显著下调冠脉内皮细胞中α-SMA和波形蛋白的表达量(P0.01),上调E-钙黏蛋白的表达量(P0.01),同时抑制Smad2和Smad3蛋白的磷酸化(P0.01),逆转TGF-β1诱导的内皮-间充质转化。结论:内皮-间充质转化过程中c-SKI表达下调,而过表达c-SKI能够抑制冠脉内皮细胞增殖及内皮-间充质转化,其机制可能与调控TGF-β1/Smad信号通路活性有关。     

慢病毒介导的SEMA3G过表达对人胰腺癌细胞系PANC-1的作用

目的通过体外实验探讨过表达SEMA3G对人胰腺癌细胞PANC-1细胞增殖、侵袭和迁移能力的影响。方法将携带SEMA3G的慢病毒载体转染人胰腺癌PANC-1细胞系,利用real-time PCR和Western blot分别检测mRNA和蛋白质水平。CCK-8法检测细胞增殖,transwell实验检测细胞侵袭和细胞划痕试验检测细胞的迁移。结果成功建立稳定转染SEMA3G胰腺癌PANC-1细胞系。SEMA3G病毒感染组与空白对照组和阴性对照组相比细胞的增殖能力显著降低(P0.05),SEMA3G病毒感染组的细胞侵袭和迁移能力明显低于两组对照组(P0.05)。结论 SEMA3G慢病毒载体能有效过表达人胰腺癌PANC-1细胞中内源性SEMA3G蛋白,进而抑制细胞增殖、侵袭和迁移。     

下调长链非编码RNA PVT1表达对胰腺癌BxPC-3细胞凋亡、侵袭及转移的影响

目的研究下调长链非编码RNA PVT1的表达对胰腺癌细胞增殖、凋亡及侵袭转移能力的影响。方法体外转染PVT1干扰序列si PVT1-1和si PVT1-2降低胰腺癌细胞系Bx PC-3 PVT1表达,同时转染阴性对照si PVT1-NC作为对照组。CCK-8实验检测细胞增殖,流式细胞术检测细胞凋亡,细胞划痕实验和Transwell实验检测细胞侵袭转移;qRT-PCR检测E-cadherin、N-cadherin、β-catenin、vimentin和PVT1 mRNA表达;Western blot检测凋亡相关蛋白(Bax、Bcl-2、caspase-3)和上皮间质转化相关蛋白(E-cadherin、N-cadherin、β-catenin、vimentin)表达。结果下调胰腺癌细胞Bx PC-3 PVT1表达能明显抑制细胞增殖、促进细胞凋亡和减少细胞侵袭转移数目(P0.05);E-cadherin蛋白和分子水平升高,N-cadherin、β-catenin、vimentin蛋白和分子水平降低(P0.05);Bax和caspase-3蛋白水平升高,而Bcl-2蛋白水平降低(P0.05)。结论下调长链非编码RNA PVT1表达能抑制胰腺癌细胞增殖和侵袭转移能力,促进胰腺癌细胞凋亡,且能够逆转胰腺癌细胞上皮间质转化。     

羧胺三唑乳清酸盐对人胰腺癌细胞系增殖及脂肪酸合成代谢的影响

目的 研究羧胺三唑乳清酸盐(CTO)对人胰腺癌细胞增殖影响及对其脂肪酸合成代谢的调控作用。方法 以人胰腺癌细胞系AsPC-1、AsPC-1/GEM(简称AR)、PANC-1、MiaPaCa-2为研究对象,用磺酰罗丹明B(SRB)检测细胞存活率,采用qPCR检测胰腺癌细胞系中脂肪酸合成关键基因mRNA水平,用Western blot检测细胞内腺苷单磷酸依赖蛋白激酶(AMPK)和乙酰辅酶A羧化酶(ACC)通路蛋白表达;利用液相色谱-质谱联用代谢组学技术检测细胞内脂质代谢物质差异。结果 与对照组相比,CTO显著降低AsPC-1、AR、PANC-1、MiaPaCa-2 4株人胰腺癌细胞活率(P<0.05); CTO下调细胞内脂肪酸合成关键基因的mRNA水平(P<0.05);CTO下调细胞中AMPK、ACC及c-Myc蛋白表达(P<0.05),而增加p-AMPK、p-ACC蛋白表达(P<0.05);CTO减少AR细胞中脂质代谢物含量(P<0.05)。结论 CTO通过抑制癌基因c-Myc蛋白表达及AMPK/ACC通路,下调脂肪酸合成相关基因的表达活性,减弱细胞脂肪酸合...     

miR-1247-5p对Smad2基因的靶向调控作用

目的利用miR-1247-5p过表达模拟物mimics,研究miR-1247-5p对TGF-β信号通路中关键蛋白Smad2的靶向调控作用。方法合成模拟miR-1247-5p过表达的模拟物mimics,及抑制miR-1247-5p表达的抑制物inhibitors;同时构建与miR-1247-5p互补靶基因Smad2的3'非翻译区,将其克隆到线性化的Report荧光报告载体中。将测序鉴定阳性的report-Smad2 3'-UTR重组质粒,与miR-1247-5p的mimics/inhibitors共转染至HEK-293T细胞,通过relative luciferase activity实验验证靶向关系,并通过Western blot实验进一步检测Smad2蛋白表达水平。结果成功构建report-Smad2 3'-UTR重组质粒,Western blot实验表明转染miR-1247-5p mimics组中Smad2蛋白表达下调(P0.05);而转染miR-1247-5p inhibitors组中,Smad2蛋白表达上调(P0.05)。结论证实miR-1247-5p直接靶向TGF-β信号通路中关键蛋白Smad2的表达,在蛋白水平对其进行调控,为进一步研究miRNA在细胞通路中的调控作用提供了依据。     

IL-9通过TGF-β/Smad通路诱导胃癌MKN-45细胞上皮-间充质转化

目的:研究白细胞介素9(IL-9)通过激活转化生长因子β(TGF-β)/Smad信号通路促进人胃癌MKN-45细胞上皮-间充质转化(EMT)的机制。方法:将常规培养的MKN-45细胞分为对照组和IL-9刺激组,利用CCK-8检测细胞活力;光镜下观察细胞形态的变化; RT-qPCR和Western blot检测EMT相关蛋白、TGF-β、Smad3和Smad5 mRNA和蛋白表达的变化; Transwell法检测细胞侵袭能力;免疫荧光染色检测细胞中波形蛋白(vimentin)的表达。结果:IL-9可以提高MKN-45细胞的存活率,且具有时间依赖性; IL-9刺激组关键转录因子Snail与Slug的表达上调,EMT相关蛋白N-cadherin、fibronectin、collagen I和vimentin表达上调,E-cadherin表达下调(P 0. 05)。Transwell结果显示IL-9组细胞侵袭能力增强,镜下观察迁移运动的细胞形态呈长梭形,RT-qPCR和Western blot结果显示TGF-β、Smad3和Smad5表达升高(P 0. 05)。结论:IL-9可以促进MKN-45细胞EMT,其作用与TGF-β/Smad信号的激活有关。     

RNA干扰环指蛋白2基因抑制胰腺癌细胞系PANC-1增殖和迁移

目的研究沉默环指蛋白2(RNF2)基因对人胰腺癌细胞系PANC-1增殖、迁移、周期和凋亡的影响及可能机制。方法用siRNA-RNF2转染PANC-1细胞沉默RNF2表达,同时设立转染无义序列(siRNA-NC)的空转染组及不进行任何处理的空白对照组(mock)。荧光定量PCR检测RNF2 mRNA表达;Western blot检测RNF2和p53表达;MTS实验和划痕实验分别检测细胞增殖和迁移能力;流式细胞计量术检测细胞转染率、凋亡率和细胞周期。结果相比正常胰腺导管上皮细胞,RNF2在胰腺癌细胞系中高表达(P<0.05)。转染siRNA-RNF2的PANC-1细胞与阴性对照组比较,RNF2 mRNA及蛋白表达下调;细胞增殖被抑制(P<0.05);细胞迁移能力下降(P<0.05);细胞凋亡率增高(P<0.05);G_0/G_1期细胞比例上升,S期和G2/M期细胞比例则降低(P<0.05)。此外,转染siRNA-RNF2显著下调PANC-1细胞中p53蛋白的表达。结论 siRNA-RNF2特异性下调胰腺癌细胞RNF2表达,显著抑制细胞增殖和迁移,结果提示RNF2可能成为胰腺癌基因治疗的一个新靶点。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

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.     

HLA in North Colombia Chimila Amerindians

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.     

Renal dysplasia and asplenia in two sibs

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.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

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.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

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.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.