环氧合酶抑制剂NS-398增强放射诱导的肝癌细胞凋亡

目的：探讨环氧合酶-2（COX-2）选择性抑制剂NS-398对放射诱导的人肝癌细胞HepG2凋亡的影响及可能作用机制。 方法:应用MTT 法检测NS-398对细胞的抑制率;透射电子显微镜观察细胞凋亡的形态学变化,流式细胞术(FCM)定量检测细胞凋亡;实时荧光定量PCR检测凋亡相关基因bcl-2、bax及caspase-3 mRNA表达;Western blotting检测Bc1-2、Bax蛋白表达;比色法分析caspase-3酶活性变化。 结果:NS-398对HepG2细胞的生长抑制作用呈时间与剂量依赖性;电镜下处理组细胞呈现典型的凋亡形态学变化,NS-398显著增加放射诱导的细胞凋亡,上调bax mRNA、Bax蛋白及caspase-3mRNA 表达,并增强caspase-3酶活性,而Bcl-2 表达无明显变化（P>0.05）。 结论:NS-398能增加放射诱导的HepG2细胞凋亡,其机制可能与上调Bax、 caspase-3表达,上升Bax／Bcl-2比例,激活线粒体凋亡通路,活化caspase-3,最终诱导细胞凋亡相关。     

华蟾素诱导膀胱癌细胞凋亡的实验研究

目的:探讨华蟾素对人膀胱癌细胞株BIU87细胞凋亡的影响.方法:采用MTT法测定华蟾素对体外膀胱癌细胞的抑制作用,倒置显微镜、TUNEL法观察细胞形态变化,用流式细胞仪检测细胞凋亡率和细胞周期的变化;半定量RT-PCR法检测凋亡相关基础Bcl-2、bax、caspase-3mRNA表达的变化,比色法测定caspase-3活性变化.结果:MTT法显示,华蟾素对膀胱癌细胞有显著的增殖抑制作用与华蟾素的浓度和作用时间有关,0.2 mg/L华蟾素作用72 h可以杀死50%以上的细胞;可诱导BIU87细胞凋亡,细胞周期停滞在S-和G2期;凋亡相关基因Bcl-2表达减少,bax无明显变化,caspase-3表达及活性增高.结论:华蟾素在体外有较强的抗膀胱癌作用,细胞凋亡的发生可能与细胞周期阻滞,Bcl-2和caspase-3基因表达及活性变化有关.     

白藜芦醇诱导胃癌原代细胞凋亡(英文)

目的：探讨白藜芦醇诱导胃癌原代细胞发生凋亡的可能性，揭示该凋亡发生与Bcl-2和Bax之间的关系。 方法： 在体外实验中，采用MTT比色法测定白藜芦醇对胃癌原代细胞的生长抑制率；以透射电镜和TUNEL 染色法，定性、定量地研究白藜芦醇与胃癌原代细胞凋亡的关系；通过免疫组织化学法和RT-PCR检测凋亡相关基因bcl-2和bax的表达。 结果： 白藜芦醇对胃癌原代细胞生长有明显抑制作用，随白藜芦醇浓度增加和培养时间的延长而增强；白藜芦醇诱导胃癌原代细胞出现典型的凋亡细胞形态；TUNEL 染色法可见，经白藜芦醇处理24 h、48 h、72 h、96 h后，胃癌原代细胞凋亡数明显随时间增加。免疫组化发现经白藜芦醇处理24 h、48 h、72 h、96 h后，胃癌原代细胞的Bcl-2蛋白阳性率减少，Bax蛋白阳性率增加。经白藜芦醇处理24、48、72、96 h后，RT-PCR检测发现胃癌原代细胞bcl-2和bax的mRNA表达阳性，并显示bcl-2 mRNA条带密度随时间的延长而递减，bax mRNA条带密度随时间的延长而增强。 结论： 白藜芦醇可能通过下调bcl-2的表达和上调bax的表达而诱导胃癌原代细胞发生凋亡。     

氧化苦参碱对人卵巢癌HO-8910细胞的作用及其机制的初步研究

目的探讨氧化苦参碱（Oxymatrine,OM）在体外诱导人卵巢癌HO-8910细胞的凋亡作用及其可能的机制。方法以不同浓度的OM作用于人卵巢癌HO-8910细胞,应用MTT法检测细胞生长情况;荧光染色观察细胞核的形态学改变;琼脂糖凝胶电泳分析细胞DNA变化;Western blot检测细胞Bcl-2和Bax的蛋白表达情况。结果 OM能显著抑制HO-8910细胞的增殖作用,具有浓度和时间依赖性。荧光染色后可观察到典型的凋亡小体。细胞DNA电泳后呈现出凋亡细胞典型的DNA ladder。Bcl-2蛋白表达量逐渐降低,Bax蛋白表达量逐渐增加,呈明显的浓度依赖性。结论 OM能诱导体外培养的人肝癌HO-8910细胞凋亡。Bcl-2表达增多,Bax表达减少是OM诱导HO-8910细胞凋亡的可能机制之一。     

As2O3诱导胃癌细胞凋亡及细胞骨架改变病理学观察

目的观察As2O3对胃腺癌细胞株SGC-7901凋亡的诱导作用及凋亡细胞骨架的改变.方法选用不同剂量As2O3处理SGC-7901细胞,分不同时问组,应用MTT法、光镜、透射电镜、TUNEL法和FCM法检测As2O3对细胞生长影响及对凋亡的诱导作用,应用荧光染色在激光共聚焦显微镜下观察凋亡细胞F-actin和α-tubulin的结构.结果As2O3抑制该细胞株的生长,MTT法显示抑制程度具有时间和剂量效应关系.光镜及透射电镜观察发现2μmol/LAs2O3作用96 h后细胞数明显减少,并出现典型的凋亡形态.TUNEL法显示2 μmol/L As2O3作用后,凋亡细胞阳性率随时间延长而增加(P＜0.05),作用96h的阳性率为(7.33±1.83)%.FCA法显示2 μmol/LAs2O3作用96 h后细胞的凋亡率为8.48%,并出现凋亡峰.激光共聚焦显微镜观察显示在细胞凋亡过程中,F-actin和α-tubulin均发生解聚和重排.结论As2O3对胃癌细胞株SGC-7901具有抑制生长及诱导凋亡的作用.微丝和微管的改变在凋亡形态变化中起着重要的作用.     

低频超声对人恶性黑色素细胞A375和MV3增殖和凋亡的影响

目的研究低频超声对人恶性黑色素细胞A375和MV3增殖和凋亡及凋亡相关基因Bcl-2表达影响。方法低频超声辐照恶性黑色素细胞A375和MV3,MTT法检测A375和MV3细胞增殖,Annexin V-FITC/PI法检测细胞凋亡,Western blot法检测凋亡相关蛋白Bcl-2的表达。结果与正常对照组相比,A375细胞和MV3细胞低频超声照射组细胞增殖显著受到抑制(P0.05),细胞凋亡明显增加(P0.05),Bcl-2蛋白表达水平明显降低(P0.05);而A375低频超声照射组和MV3低频超声照射组之间没有显著性差异。结论低频超声可用于恶性黑色素细胞瘤的治疗。     

XAV939对卵巢癌细胞增殖的抑制作用

目的观察XAV939对卵巢癌细胞株OVCAR3增殖的作用。方法采用MTT法检测XAV939对卵巢癌细胞增殖的抑制作用,用流式细胞技术检测卵巢癌细胞株OVCAR3细胞周期和凋亡,Western blotting法检测Cyclin D1、Bcl-2的表达以及PARP剪切片段。结果 XAV939对卵巢癌细胞株增殖的抑制呈现浓度依赖性和时间依赖性,细胞周期检测表现为G1期阻滞,凋亡检测显示凋亡率增加。不同浓度的XAV939处理组的卵巢癌细胞Cyclin D1、Bcl-2蛋白表达水平下降,PARP剪切片段增多。结论 XAV939可以抑制卵巢癌细胞株OVCAR3的增殖。     

苦参素诱导卵巢癌HO8910细胞凋亡的实验研究

目的探讨苦参素对卵巢癌细胞株HO8910的抑制增殖效应及凋亡诱导作用。方法用不同浓度苦参素处理HO8910细胞,采用MTT法检测药物对细胞的抑制作用,倒置显微镜和电镜观察细胞形态学改变;以及应用流式细胞仪观察细胞凋亡过程中细胞周期的变化。结果随着药物浓度的升高,作用时间的延长,苦参素对HO8910细胞的生长抑制率逐渐增高;在苦参素作用下,HO8910细胞呈凋亡改变,出现凋亡小体。细胞凋亡的同时,细胞周期发生特定的改变,亚G1峰出现,S+G2～M期细胞所占比例组明显增高。结论苦参素能诱导卵巢癌细胞凋亡,抑制卵巢癌细胞增殖。     

乙型肝炎病毒I97L变异核壳蛋白对诱导HepG2细胞凋亡的影响

目的研究197L变异核壳蛋白对诱导HepG2细胞凋亡的影响。方法构建EGFP-野毒株核壳蛋白、197L变异核壳蛋白融合表达载体（pEGFP—WT和pEGFP—L97），酶切和测序鉴定；将pEGFP—WT、pEGFP—L97和对照质粒分别转染HepG2细胞．筛选阳性细胞株；荧光显微镜观察阳性克隆细胞荧光蛋白表达，Western—blot检测核壳蛋白表达：以TNF-α、Act-D诱导HepG2细胞株凋亡，0、16、32和48h采用流式细胞术检测细胞凋亡，32h时采用共聚焦检测细胞凋亡比例。结果成功建立了融合表达蛋白表达载体；荧光显微镜显示各细胞克隆有较好的荧光蛋白表达，Western-blot检测各细胞系核壳蛋白表达没有差异；16、32、48h时，pEGFP-WT、pEGFP-L97表达细胞株凋亡率均明显低于pEGFP-C1细胞株（P〈0．05）；32h和48h时，pEGFP-L97细胞株凋亡率明显高于pEGFP-WT细胞株（P〈0．05）；32h时激光共聚焦检测结果与流式细胞术检测结果一致。结论乙型肝炎病毒197L变异核壳蛋白对诱导HepG2细胞凋亡的影响与野毒株核壳蛋白不同，与野毒株核壳蛋白相比，197L变异核壳蛋白对凋亡因素可能更敏感。     

As2O3诱导喉鳞癌Hep-2细胞凋亡及其机制的研究

目的 探讨三氧化二砷( As2O3)诱导喉鳞癌Hep-2细胞凋亡及其作用机制.方法 体外培养喉鳞癌Hep-2细胞株,MTT法检测不同作用时间及不同浓度的As2O3对Hep-2细胞生长的作用;TUNEL染色、电镜观察Hep-2细胞凋亡的发生和形态学改变;Western印迹检测As2O3诱导Hep-2细胞凋亡时Survivin蛋白的表达改变.结果 As2O3可明显抑制喉鳞癌Hep-2细胞的生长(P＜0.05),并随药物浓度及作用时间的增加而增强.TUNEL、电镜结果显示As2O3作用后,Hep-2细胞发生凋亡.As2O3作用后,Survivin基因的表达降低.结论 As2O3可诱导喉鳞癌Hep-2细胞凋亡,其生长抑制作用呈时间及剂量依赖性;其机制可能是As2O3通过抑制凋亡抑制因子Survivin蛋白的表达,促进细胞凋亡,从而发挥抗肿瘤的作用.     

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.     

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.     

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.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'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.