齐墩果酸诱导人白血病HL-60细胞凋亡及细胞周期阻滞

目的：探讨齐墩果酸诱导HL-60细胞凋亡作用及其对细胞周期的影响。方法：以HL-60细胞为研究对象，以不同剂量的齐墩果酸处理HL-60细胞12 h、24 h、48 h、72 h后，用MTT法观察HL-60细胞的生长抑制率；琼脂糖凝胶电泳测定DNA梯形条带；流式细胞仪分析HL-60细胞细胞周期的变化；同时用Western blotting 检测凋亡途径最终执行者caspase-3的表达。结果：齐墩果酸对HL-60细胞生长具有明显的抑制作用，其中80 μmol/L齐墩果酸作用48 h对HL-60细胞的抑制率达到50%以上，齐墩果酸诱导细胞凋亡呈明显的时间和剂量依赖性； HL-60经齐墩果酸处理48 h出现典型的DNA梯形条带；以80 μmol/L的齐墩果酸处理HL-60细胞12 h即可以使caspase-3的前体procaspase-3断裂为有活性的caspase-3；流式细胞技术检测结果表明HL-60细胞经齐墩果酸处理后细胞周期阻滞于G₁期，其中48 h和72 h分别达到63.24%和67.90%。结论：齐墩果酸可以诱导HL-60细胞凋亡，并使细胞阻滞于G₁期。     

rhLIF与IL-24基因协同诱导HL-60细胞的凋亡

目的：构建能稳定表达白血病抑制因子（LIF）的转基因细胞,并研究所表达的LIF与IL-24基因在诱导HL-60细胞凋亡方面的协同作用。方法：用真核表达质粒pcDNA3-LIF转染ECV304细胞,G418筛选阳性细胞,收集阳性细胞和培养上清,RT-PCR检测LIFmRNA的表达。同时在已转化重组腺病毒质粒的大肠杆菌中抽提质粒pAdEasy-1-pTrack-CMV-IL-24,用PacI酶切使重组腺病毒质粒线性化,脂质体转染QBI-293A细胞,收获Ad-IL-24腺病毒子。将重组病毒子（Ad-IL-24）感染HL-60细胞,同时加入含LIF的培养上清,并设对照组,RT-PCR检测IL-24 mRNA表达,光镜下观察HL-60细胞形态的变化,激光扫描共聚焦显微镜观察细胞凋亡改变,流式细胞仪检测细胞凋亡率,免疫细胞化学分析凋亡因子的改变。结果：成功构建能稳定表达LIF蛋白的转基因细胞;获得高滴度的重组腺病毒Ad-IL-24,用它感染HL-60细胞后,能检测到IL-24 mRNA的表达。各种检测方法表明LIF、IL-24基因都能抑制HL-60细胞生长、诱导凋亡,且两者具有协同作用。结论：LIF、IL-24基因都能抑制HL-60细胞生长,诱导凋亡,两者具有协同作用。     

细胞色素C对HL-60细胞促凋亡作用及其对bcl-2、bcl-xl基因表达的影响

目的：探讨细胞色素C在体外作用于HL-60细胞时细胞发生的变化及其相关凋亡基因bcl-2、bcl-xl表达变化的机制。方法:用不同浓度的细胞色素C作用于HL-60细胞24 h，然后用MTT检测细胞色素C对HL-60细胞抑制率；用普通光镜、荧光显微镜检测HL-60细胞形态的变化；用流式细胞仪、DNA凝胶电泳对HL-60细胞凋亡的检测；用RT-PCR检测bcl-2、bcl-xl mRNA表达的变化。结果:细胞抑制率随着细胞色素C浓度的增加而增加；当细胞色素C浓度在0-37.5 mg/L作用HL-60细胞24 h，随着细胞色素C浓度的增加，HL-60细胞发生的凋亡逐渐增加，可见典型的凋亡细胞和明显的DNA梯度条带；同时，在该浓度范围内，bcl-2、bcl-xl mRNA表达逐渐减少；当细胞色素C浓度大于37.5 mg/L时，细胞凋亡率并不增加，而是下降，但是坏死细胞明显增加。结论：一定浓度细胞色素C能诱导HL-60细胞发生凋亡，并且细胞凋亡率、bcl-2、bcl-xl基因表达的变化与细胞色素C浓度呈一定的量效依赖关系，细胞色素C诱导HL-60细胞凋亡可能与抑制bcl-2、bcl-xl基因的表达有一定的关系。     

三七总皂甙诱导HL-60细胞凋亡的研究

目的 :观察三七总皂甙是否能诱导人白血病细胞株HL 60细胞凋亡及相关基因的关系。方法 :取 2 0 0～ 1 60 0ug/ml三七总皂甙处理HL 60细胞 ,光镜下观察细胞形态 ;MTT法测细胞生长抑制率 ;流式细胞仪检测细胞周期 ;免疫组化法检测 p5 3 ,bcl 2凋亡相关基因的表达 ,采用真彩色图像分析仪定量分析免疫组化的结果 ;其结果表明 2 0 0～ 1 60 0ug/ml三七总皂甙可抑制HL 60细胞生长 ,抑制率随着剂量的增加而增加。流式细胞仪检测凋亡峰随药物浓度增大而增加。80 0 ,1 60 0ug/毫升三七总皂甙可使 p5 3基因表达上调 ,bcl 2表达明显下降 ,这两种基…     

硫化砷诱导HL-60细胞凋亡中端粒酶活性和hTERT mRNA表达的改变

目的：探讨硫化砷在诱导急性非淋巴细胞白血病(APL)细胞株HL-60凋亡的同时，对端粒酶活性的影响及作用机制。方法：采用PCR—ELISA法测定端粒酶活性；半定量RT—PCR检测hTERT mRNA的表达；流式细胞仪分析细胞周期、细胞凋亡及CD11b表达。结果：用0．3～0．6mg/L的硫化砷作用72h，不能诱导HL-60细胞凋亡，将剂量增至1．5～3．0mg/L时，凋亡细胞的比率逐渐增高，同时伴随HL-60细胞端粒酶活性和hTERT mRNA表达受抑。随着硫化砷浓度的增高，HL—60细胞在G2／M期的比率渐增高。3．0mg/L的硫化砷作用72h，HL-60细胞CD11b的表达由1．27％升至6．07％，结论：硫化砷在诱导HL-60细胞凋亡的同时，下调其端粒酶活性。G2／M期细胞比率的增高可能与端粒酶活性降低相关。高浓度硫化砷72h可轻度诱导HL—60细胞分化。     

茶多酚对HL-60细胞株体外增殖及细胞周期的影响

目的 探讨茶多酚对人急性早幼粒白血病细胞株HL-60细胞增殖和细胞周期的影响.方法 采用四甲基偶氮唑盐微量酶反应比色法(MTT比色法)观察茶多酚对体外培养的HL-60细胞增殖活性的影响.采用HE染色、荧光染色观察用茶多酚后细胞形态变化.用流式细胞仪(FACS)检测细胞凋亡率及细胞周期.结果 (1)MTT比色法检测显示茶多酚能抑制HL-60细胞增殖,在一定范围内呈剂量和时间依赖性(P＜0.05).当茶多酚浓度达到400和800 mg/L时,48 h抑制率分别为(58.90±1.19)%和(72.57±0.70)%.(2)流式细胞仪分析,茶多酚处理组出现一特征性的亚二倍体凋亡峰.其凋亡率呈时间、剂量依赖性.(3)流式细胞术发现茶多酚可使HL-60细胞阻滞于S期,其阻滞细胞的数量与药物浓度呈正相剂量关系,以作用24 h对细胞周期的阻滞作用最强.结论 茶多酚能有效抑制HL-60细胞增殖,在一定范围内具有时间和剂量依赖性.茶多酚可体外诱导HL-60细胞凋亡,并使细胞周期阻滞于S期.     

一氧化氮诱导HL-60细胞凋亡及其机制探讨

目的：观察一氧化氮对人类白血病细胞是否具有致凋亡作用，并研究Bcl-2基因和P53基因在此过程中的作用。方法：将不同浓度的外源性一氧化氮供体亚硝基铁氰化钠与HL-60细胞作用，观察其作用的时间效应和剂量效应；用MTT法观察NO对细胞的抑制作用，用透射电镜观察细胞结构改变，用DNA凝胶电泳、细胞DNA含量、DNA末端标记、Annexin-V/PI法等分析细胞凋亡，并用流式细胞法进一步观察在NO作用过程中凋亡调控因子Bcl-2和P53蛋白及线粒体膜蛋白表达变化。结果：NO能抑制HL-60细胞生长，并在一定的剂量范围内显示作用时间和剂量的量效关系；典型的细胞形态改变、DNA片段化、亚G1峰检出、DNA末端标记、Annexin-V/PT^-表达增加等证实。NO能诱导白血病细胞凋亡；在此过程中，P53蛋白表达上调，而Bcl-2蛋白表达下调，线粒体膜蛋白表达增加。结论：NO对HL-60细胞有强的致凋亡作用，Bcl-2和P53参与NO诱导HL-60细胞凋亡的调控。     

SU11248对白血病细胞HL-60增殖及凋亡作用的实验研究

目的:探讨新型抗肿瘤药物苹果酸舒尼替尼(SU11248)对白血病细胞HL-60的生物学效应的影响及其作用机制。方法:应用MTT法检测SU11248对HL-60细胞增殖能力的影响;用AnnexinV/PI双染流式细胞术检测细胞凋亡;用流式细胞技术分析细胞的DNA倍体及细胞周期变化;用凝胶电泳分析DNA片段化;以Western blot法检测2.0μg/ml SU11248作用HL-60后bcl-2、bax、caspase-3蛋白水平的变化。结果:SU11248可明显抑制HL-60细胞增殖(P<0.05),呈剂量和时间依赖性,半数抑制浓度(IC50)约为2.00μg/ml。SU11248可促进细胞凋亡,并呈剂量依赖性;能将HL-60细胞阻滞于G0/G1期,并呈时间依赖性;诱导HL-60细胞呈典型的DNA梯带;SU11248作用后bcl-2蛋白表达随时间依赖性降低,caspase-3蛋白表达升高,bax蛋白表达无明显变化。结论:SU11248能抑制HL-60细胞增殖,诱导其凋亡,调节bcl-2家族蛋白的表达,并裂解caspase-3是其可能作用机制之一。     

干扰素对HL-60细胞与维甲酸耐药HL-60细胞作用的研究

目的:探讨干扰素α对HL-60细胞增殖分化与凋亡的作用及特点.方法:MTT法测定细胞增殖性变化.NBT方法测定细胞分化程度.流式细胞仪测定凋亡的发生程度.浓度递增法诱导HL-60细胞的维甲酸耐药性.结果:IFNα体外抑制HL-6 0细胞增殖,并与维甲酸产生协同抑制作用.IFNα可单独诱导HL-60细胞分化,又能增加维甲酸诱导分化作用.单独IFNα无明显的诱导HL-60细胞凋亡作用.维甲酸耐药HL-60细胞经IFNα作用3 d后,可明显恢复其对维甲酸的反应性.结论: IFNα既能促进维甲酸对HL-60细胞的诱导分化作用,又可逆转HL-60的维甲酸耐药性.     

白血病细胞TGF-β1含量减少及外源性TGF-β1基因对HL-60细胞的作用

目的：研究白血病细胞是否存在转化生长因子β1（TGF-β1）量的异常以及这种异常在白血病发病机制中的可能作用。 方法： 应用ELISA法检测白血病细胞株和原代白血病细胞培养上清TGF-β1水平，进一步应用脂质体介导基因转移法将TGF-β1基因转染HL-60细胞，采用有限稀释法及G418筛选得到抗性细胞，应用RT-PCR、白血病细胞集落培养、裸鼠移植瘤成瘤试验、片段化DNA分析、流式细胞术检测HL-60细胞内TGF-β1、bcl-2、端粒酶反转录酶（hTERT）mRNA的表达变化等方法了解外源性TGF-β1基因对HL-60细胞体内外增殖、凋亡的影响。 结果： 白血病细胞株和原代白血病细胞培养上清TGF-β1水平明显低于正常对照组(P<0.01)，转染TGF-β1基因后，HL-60细胞内TGF-β1 mRNA表达上调，bcl-2、hTERT mRNA表达下调，细胞体外增殖受抑制，集落形成抑制率达78.3%，裸鼠移植瘤生长受抑制，荷瘤鼠生存期延长，细胞呈现凋亡特征性的梯形条带，凋亡比例达73.4%。 结论： 内源性TGF-β1水平降低是白血病的发病机制之一,外源性TGF-β1基因能够抑制HL-60细胞增殖，诱导细胞凋亡,该基因通过下调bcl-2、hTERT mRNA的表达而发挥作用。     

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.