沉默Bax基因对TNF-α诱导人肺泡Ⅱ型上皮细胞凋亡的影响

目的:运用RNA干扰(RNAi)技术沉默人肺泡Ⅱ型上皮细胞株A549的B细胞淋巴瘤-2相关基因(Bax),探讨Bax在肿瘤坏死因子-α(TNF-α)诱导A549细胞凋亡中的作用。方法:体外培养A549细胞,利用脂质体LipofectamineTM2000将化学合成的Bax小干扰RNA(siRNA)转染入A549细胞,给予10μg/LTNF-α刺激24h,RT-PCR检测bax mRNA的表达,Western blotting和免疫组化检测Bax蛋白的表达,流式细胞术检测细胞凋亡率的变化。结果:化学合成的Bax siRNA抑制了A549细胞Bax mRNA和蛋白的表达(P0.05),流式细胞术显示BaxsiRNA组的细胞凋亡率明显低于TNF-α组和阴性对照siRNA组(P0.05)。结论:体外实验证明Bax的高表达在TNF-α诱导A549细胞凋亡中发挥了重要的促凋亡作用,利用RNAi技术沉默Bax基因可以有效抑制由TNF-α介导的A549细胞凋亡。     

二氢杨梅素对肺癌A549细胞凋亡的作用

目的 探讨二氢杨梅素（DMY）对肺癌A549细胞的诱导凋亡作用。方法 以肺癌A549细胞为研究对象,采用MTT法研究DMY对A549细胞的抗肿瘤作用,并进一步通过流式细胞术检测DMY对A549细胞的凋亡作用,免疫印迹法（Western blotting）检测促凋亡因子Bax和抗凋亡因子Bcl-2表达水平,进一步探讨其机制。
结果 DMY浓度越大,对A549细胞的抑制作用越明显,48h的IC50为64.45g/L;流式细胞术显示,随着DMY浓度的增加A549细胞凋亡也逐渐增加;Western blotting结果显示,DMY可诱导A549细胞中凋亡因子Bax蛋白表达增加,同时抗凋亡因子Bcl-2蛋白表达减少,尤其高浓度组改变明显。
结论 DMY可以在体外诱导A549细胞凋亡。     

GeXP 检测rhIL-24 联合DDP 对人肺腺癌A549 / DDP 细胞凋亡相关基因的实验研究

目的:利用多基因遗传表达分析系统(GeXP)探讨重组人白细胞介素-24(rhIL-24)联合顺铂(DDP)诱导人肺腺癌顺铂耐药株A549/ DDP 细胞6 个凋亡相关基因的变化。方法:用rhIL-24、DDP 及rhIL-24+DDP 干预A549/ DDP 细胞,应用多基因遗传表达分析系统(GeXP)同时检测Bax、Bcl-2、Survivin、Caspase3、Rb 与P53 等6 个凋亡相关基因。结果:rhIL-24 蛋白可引起A549/ DDP 细胞Bax 基因、Caspase3 基因、Rb 基因转录上调;Bcl-2 基因、Survivin 基因转录下调,但抑癌基因P53 变化无规律,rhIL-24 联合DDP 后Bax、Survivin、Rb 表达较单独rhIL-24 组变化更加显著。结论:rhIL-24 蛋白可通过上调Bax,下调Bcl-2、Survivin,激活Caspase3,上调抑癌基因Rb 诱导人肺腺癌A549/ DDP 细胞凋亡。     

β-榄香烯通过抑制HIF-1α表达促进人肺癌细胞凋亡

目的探讨β-榄香烯是否通过抑制肺癌细胞中HIF-1α的高表达促进肺癌细胞的凋亡及相关机制。方法常规培养人肺癌A549细胞,实验分为对照组、乏氧组、β-榄香烯组、不同浓度β-榄香烯/乏氧处理组;通过转染含有HIF-1α基因的质粒过表达HIF-1α基因。应用Annexin V-FITC凋亡检测试剂盒及流式细胞仪、TUNEL检测细胞凋亡。并采用免疫荧光检测HIF-1α蛋白的活化、Westen blot方法检测Bax、caspase-3及Bclx L蛋白表达及转染效率。结果β-榄香烯能够抑制肺癌A549细胞的增殖力,呈时间和浓度依赖性,能够诱导乏氧状态下的肺癌A549细胞凋亡。β-榄香烯可通过上调A549细胞的Bax、caspase-3蛋白表达,降低Bcl-xL表达来诱导乏氧状态下肺癌A549的凋亡。此外,β-榄香烯可有效降低乏氧状态下HIF-1α蛋白的高表达。通过转染过表达HIF-1α基因,大大降低了β-榄香烯的诱导凋亡作用。结论β-榄香烯可以上调乏氧状态下A549细胞凋亡,可能与抑制HIF-1α的高表达相关。     

芹菜素对人肺癌A549细胞凋亡及相关蛋白Bax、Bcl-2表达的影响

目的研究芹菜素对人肺癌 A549细胞增殖抑制和凋亡诱导作用与 Bax、Bcl-2蛋白表达的影响。方法10~80μmol/L 不同浓度芹菜素作用 A549细胞,采用 MTT 法检测芹菜素对 A549细胞增殖抑制作用;Hoechst 33258细胞核染色法观察芹菜素诱导细胞凋亡形态学的变化;流式细胞仪 AnnexinV- FITC/PI 双染色法检测细胞凋亡率;Western blot 法检测凋亡相关蛋白 Bax 和 Bcl-2表达的变化。结果 MTT 法显示,芹菜素对 A549细胞有显著的增殖抑制作用（P <0.01）,且具浓度和时间依赖性;荧光显微镜下观察到芹菜素处理组细胞出现典型的凋亡形态学改变：细胞核固缩、染色质凝集和核碎片化等;流式细胞仪分析结果显示,芹菜素呈浓度依赖性诱导 A549细胞凋亡;Western blot结果显示,促凋亡蛋白 Bax 随着芹菜素浓度升高表达增加,抗凋亡蛋白 Bcl-2随着芹菜素浓度升高表达减少。结论芹菜素具有抑制人肺癌 A549细胞增殖,诱导其凋亡的作用,其机制可能与上调 Bax 蛋白表达和下调 Bcl-2蛋白表达有关。     

呼吸道合胞病毒感染对细胞凋亡及FasL、Fas、Bcl-2、Bax基因表达的研究

目的：研究呼吸道合胞病毒(RSV)感染与细胞凋亡的关系及对凋亡相关基因FasL、Fas、bcl-2和bax表达的影响。 方法： 采用A549细胞，在RSV感染后不同时点收集细胞，流式细胞仪和透射电镜检测细胞凋亡，免疫组织化学法检测凋亡相关基因FasL、Fas、Bcl-2和Bax表达情况。 结果： 流式细胞仪检测结果RSV感染后72 h（6.61%）、120 h（10.94%）的细胞凋亡指数明显高于对照组（4.32%、5.31%）；免疫组化法检测结果对照组FasL、Bax基因呈现无表达或局部弱表达；随着RSV感染时间的延长，bax、 Fas、 FasL基因的表达均高于对照细胞，bcl-2基因呈现弱表达或无表达。 结论： RSV在感染后期能诱导宿主细胞发生凋亡，促凋亡基因FasL、Fas 、Bax和抗凋亡基因Bcl-2表达水平的差异是RSV诱导凋亡的机制之一。     

pcEgr-hp53稳定转染联合X射线诱导人肺腺癌A549细胞凋亡及相关凋亡蛋白表达变化

目的：探讨辐射诱导表达载体pcEgr-hp53体外稳定转染人肺腺癌A549细胞后联合X射线照射,诱导细胞凋亡的作用及相关凋亡蛋白Bax、Bcl-2和caspase-3蛋白表达的变化。方法：以脂质体介导携有外源野生型p53基因的辐射诱导表达载体pcEgr-hp53和pcDNA3.1,体外转染A549细胞,筛选稳定转染的细胞克隆并扩增培养,所表达的A549-hp53和A549-vect分别接受0、0.5、2和5Gy X射线照射,即8个实验组,采用TUNEL法和流式细胞术检测稳定转染联合辐射对细胞凋亡和Bax、Bcl-2和caspase-3蛋白表达变化的影响。结果：A549-hp53组加不同剂量照射与0 Gy组比较,其百分数均明显增加（0.5-5Gy,P〈0.05-P〈0.01）,Bcl-2蛋白表达在0.5-5Gy时明显下降,而Bax蛋白明显增加（P〈0.05-P〈0.01）,caspase-3蛋白表达在2-5Gy时明显增加（P〈0.01）;A549-hp53照射组不同剂量照射,与A549-vect相应照射剂量组比较,其百分数在0.5Gy时无明显差异,在2-5 Gy时为（P〈0.01）,Bcl-2蛋白表达明显下降（0.5Gy,P〈0.01;2-5Gy,P〈0.05）,Bax和caspase-3蛋白表达0.5-5Gy时明显增加（P〈0.05-P〈0.01）。结论：体外p53基因转染联合X射线照射可诱导肿瘤细胞凋亡明显增多,凋亡相关蛋白Bcl-2表达下调,Bax、caspase-3表达上调,具有显著的肿瘤抑制作用。     

miR-221 靶向AdipoR1 基因对LPS 诱导的肺泡上皮细胞A549 炎症分泌及凋亡影响

目的:探讨miR-221是否靶向脂联素受体1 AdipoR1基因影响脂多糖(LPS)诱导的肺泡上皮细胞A549炎症分泌和凋亡。方法:随机将人肺泡上皮细胞A549分为对照组、LPS组和LPS+转染组,RT-qPCR法检测miR-221和AdipoR1 mRNA表达,Western blot法检测AdipoR1蛋白及凋亡相关因子Bax、Bcl-2蛋白表达,流式细胞仪检测细胞凋亡率,ELISA法检测细胞培养上清液中炎性因子IL-1β、IL-6和肿瘤坏死因子-α(TNF-α)浓度,双荧光素酶报告基因检测miR-221是否靶向AdipoR1。结果:与对照组比较,LPS组细胞中miR-221及Bax蛋白表达升高(P0.05),AdipoR1 miRNA和蛋白及Bcl-2蛋白表达降低(P0.05),细胞培养上清液中IL-1β、IL-6、TNF-α浓度升高(P0.05),细胞凋亡率升高(P0.05);双荧光素酶报告基因实验证实miR-221靶向负调控AdipoR1的表达;抑制miR-221和过表达AdipoR1均能抑制LPS诱导肺泡上皮细胞分泌炎症因子,抑制细胞凋亡;抑制AdipoR1逆转了抑制miR-221对LPS诱导的肺泡上皮细胞炎症分泌和凋亡的影响。结论:miR-221靶向AdipoR1影响LPS诱导的肺泡上皮细胞A549炎症分泌及凋亡。     

外源性FHIT基因表达对肺腺癌细胞株A549恶性表型的逆转作用

目的 探讨FHIT（fragile histidine triad)基因表达在肺癌细胞增殖、凋亡、成瘤性中的作用。方法 应用脂质体转染法构建能高效表达外源性FHIT基因的A549－FHIT细胞和作为空载体对照的A549－vector细胞，裸鼠皮下接种构建肺癌移植瘤动物模型，应用逆转录－PCR、Western杂交、免疫组化、流式细胞计数等技术检测外源性FHIT基因转录、表达及对肺癌细胞株增殖、凋亡、成瘤性等的影响。结果 A549－FHIT细胞的生长曲线、克隆形成率、移植瘤体积和重量均显著低于A549－vector和亲本A549细胞；A549－FHIT细胞凋亡水平显著高于A549－vector和亲本A549细胞；与两个对照组相比，A549－FHIT细胞G0／G1比例显著增高。结论 外源性FHIT表达基因能够显著抑制肺癌细胞增殖和分裂，诱导其凋亡，抑制其成瘤性，提示FHIT基因在肺癌中具有抑癌基因的功能。     

miR-29b通过调控Bax基因抑制LPS诱导的肺泡上皮细胞系A549凋亡

目的探讨miR-29b对脂多糖(LPS)诱导的肺泡上皮细胞系A549凋亡的影响及其机制。方法将体外培养的A549细胞分为对照组、LPS组(给予10 mg/L的LPS处理)、LPS+miR-NC组(转染miR-29b mimics阴性对照后给予LPS处理)、LPS+miR-29b组(转染miR-29b mimics后给予LPS处理);用RT-qPCR检测细胞中miR-29b的表达水平;MTT法检测细胞存活率;流式细胞仪检测细胞凋亡率;Western blot检测Bcl-2、Bax和cleaved caspase-3蛋白的表达;双荧光素酶报告基因实验检测Bax和miR-29b的靶向关系。结果与对照组相比,LPS组、LPS+miR-NC组和LPS+miR-29b组细胞中miR-29b、Bcl-2蛋白的表达水平和细胞存活率均明显降低,而细胞凋亡率和Bax、cleaved caspase-3蛋白的表达水平均明显升高(P<0.05);与LPS组相比,LPS+miR-29b组细胞中miR-29b、Bcl-2蛋白的表达水平和细胞存活率均明显升高,而细胞凋亡率和Bax、cleaved caspase-3蛋白的表达水平均明显降低(P<0.05)。双荧光素酶报告基因实验证实Bax是miR-29b的潜在靶基因。结论miR-29b可抑制LPS诱导的A549细胞凋亡,其作用机制可能与靶向调控Bax表达有关。     

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.     

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 dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ 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 $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ 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 $\text{primates}$. 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.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

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.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

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.     

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.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

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.     

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.