1型糖尿病、2型糖尿病及2型糖尿病伴动脉硬化患者外周血白细胞端粒长度的研究

目的:检测1型糖尿病(T1DM)、2型糖尿病(T2DM)、2型糖尿病伴动脉硬化(DAS)患者外周血白细胞端粒长度,分析糖尿病患者端粒长度变化的因素。方法:选择T1DM患者30例、T2DM患者60例、DAS患者40例和健康对照(NC组)40例,分别提取外周血白细胞,然后提取基因组DNA进行real-time PCR检测端粒长度。利用多元线性回归分析影响端粒长度变化的因素。结果:T1DM组、T2DM组和DAS组的端粒长度均小于NC组(P0.05);T1DM组端粒长度短于T2DM组和DAS组;DAS组较T2DM组更短。多元线性回归分析显示,T1DM组中,年龄与端粒长度呈负相关(P0.05);T2DM组中,年龄、体重指数(BMI)与端粒长度呈负相关(P0.05);DAS组中,患病时间、BMI与端粒长度呈负相关(P0.05)。结论:糖尿病患者的外周血白细胞端粒长度明显短于正常人,并且T1DM患者端粒长度短于T2DM;在2型糖尿病的对比中,DAS患者的端粒长度明显短于T2DM。患者的年龄、患病时间、BMI与端粒长度的缩短有密切的关系。     

大肠癌中端粒长度与DCC基因mRNA表达的研究

目的 分析端粒长度及DCC基因mRNA表达在大肠癌及腺瘤发生发展中的作用。方法 采用Sourthern印迹杂交及RT-PCR技术,分别检测46例大肠腺瘤及62例大肠癌组织中的端粒限制性片段（TRF）长度及DCCmRNA表达状态并观察它们与肿瘤临床病理的关系。结果 在大肠癌及大肠腺瘤中,TRF长度较正常组织明显缩短,其缩短者占53.2%和41.3%,而延长者仅占6.5%和4.4%,结肠癌的TRF长度也较直肠癌的TRF长度明显缩短,DCCmRNA表达缺失率在大肠癌及大肠腺瘤中则分别达62.9%和34.8%,显著高于正常组织（1.6%);同时,大肠癌患者的平均TRF长度还随患者年龄的增长而缩短,DCCmRNA表达缺失率则随肿瘤的分化程度下降及临床阶段的进展而升高,但DCCmRNA表达缺失与TRF长度缩短在大肠癌中未表现出明显的相关性。结论 端粒缩短与DCCmRNA表达缺失与TRF长度缩短在大肠癌中未表现出明显的相关性。结论 端粒缩短与DCCmRNA表达缺失可能是大肠腺瘤恶变及大肠癌形成过程中较具特征表现的生物学异常行为。     

6A8α-甘露糖苷酶表达对人鼻咽癌细胞CNE-2L2端粒酶活性和端粒长度的影响

探讨端粒长度与端粒酶活性在人鼻咽癌细胞CNE-2L2恶性行为改变前后的变化，建立研究恶性行为改变与端粒长度与端粒酶活性间关系的细胞模型。与6A8α-甘露糖苷酶表达正常的CNE-2L2细胞(野生型细胞W，转导空载体的细胞M及转导无关DNA片段的细胞S)相比，6A8α-甘露糖苷酶表达低下的细胞(AS)接种裸鼠皮下后的肿瘤性生长受抑。用Telo TAGGG Telomere Length Assay Kit及Telomerase PCR ELISA Kit分别测定端粒长度及端粒酶活性，用RT-PCR方法分析端粒重复序列结合因子(TRF)的转录水平。见AS细胞的端粒明显缩短(6．78Kb，W细胞为8．40Kb，M细胞为8．34kb，S细胞为9．56kb)，但端粒酶活性及端粒重复序列结合因子l和2(TRFl和2)的转录水平未见改变。实验表明，恶性行为降低的CNE-2L2细胞的端粒变短，但与端粒酶活性及TRF1／2无关，提示在CNE-2L2细胞中可能存在着端粒酶及TRF1／2以外的调节端粒长度的机制。这为研究肿瘤细胞恶性行为改变与端粒长度与端粒酶活性间关系提供了一个模型。     

人骨髓间质干细胞中的端粒调控机制

目的：〖HT5"SS〗研究人骨髓来源间充质干细胞(MSCs)端粒长度的调控机制。方法：以贴壁培养法从人骨髓中分离MSCs并用MSCs及造血干细胞相关表面抗体作表型鉴定,用Southern blotting检测MSCs的端粒长度;应用免疫荧光染色技术检测端粒重复序列结合因子1（TRF1）和早幼粒细胞白血病蛋白小体（PML）的定位;以端粒重复序列扩增法（TRAP）和/或Western blotting法检测传代及分化成脂肪细胞的MSCs和经同步化处理被阻断在S期的MSCs的端粒酶表达。结果：与端粒酶阴性ALT细胞株WI-38-2RA细胞相比,MSCs的端粒长度较短并且端粒长度变异度不大;端粒调控相关蛋白TRF1和PML在MSCs中的定位则与端粒酶阳性细胞HeLa细胞相同,两者呈非共定位,而在端粒酶阴性WI-38-2RA细胞中两者呈共定位状态。MSCs中不存在有染色体外端粒重复序列DNA（ECTR DNA）。TRAP法检测传代培养的MSCs端粒酶呈阴性表达,但分化成脂肪的MSCs端粒酶呈阳性表达。Western blotting 检测同步化处理前MSCs端粒酶呈微弱表达,经同步化处理被阻断在S期时,MSCs的端粒酶表达明显增高,并且与S期的细胞比例呈正相关。结论：MSCs中不存在ALT相关的早幼粒细胞白血病蛋白小体(APBs)、染色体外端粒重复序列DNA(ECTR DNA)和端粒长度较长、端粒长度变异度大等ALT机制相关分子特征;非同步化在S期处理的MSCs,端粒酶呈微弱表达,但诱导向脂肪细胞分化或处在S期时,MSCs的端粒酶表达明显增高,并且与S期的细胞比例呈正相关。本研究提示MSCs是通过端粒酶机制而不是端粒延长旁路途径（ALT）机制调控其端粒末端。     

乳腺良恶性病变组织端粒长度和端粒酶活性检测

目的　比较乳腺良恶性病变端粒长度改变及其在肿瘤发生发展中的意义 ;探讨端粒酶活性与临床病理参数的关系及其在乳腺癌诊断中的价值。方法　Southern印迹杂交检测TRF长度 ,端粒重复扩增分析 (TRAP)方法检测端粒酶活性。结果　乳腺癌组织平均TRF为 (5 2± 2 8)kb ,与正常组织比较明显缩短 (P <0 0 0 1) ,从正常乳腺组织到乳腺良性病变、乳腺原位癌及浸润性癌平均TRF呈递减趋势。 5 8例乳腺癌中 4 9例端粒酶阳性 (84 7% ) ,端粒酶活性与临床病理参数无相关性 ;癌旁组织端粒酶为阴性 ,而 7例乳腺增生症和 6例乳腺纤维腺瘤中分别有 1例端粒酶阳性 ,与乳腺癌比其差异有显著性 (P <0 0 0 1)。结论　端粒长度在肿瘤发生发展过程中渐进性缩短 ,并最终触发端粒酶的激活 ;端粒酶活性检测有望成为乳腺癌诊断的可靠标记物     

端粒长度与卵巢功能的相关性研究

目的探索端粒长度变化与女性卵巢功能的关系及其临床意义。方法在本院2006.3～2008.7住院治疗的各个年龄段的育龄女性及卵巢早衰患者正常卵巢组织标本66例,常规酚一氯仿法提取组织DNA后与端粒寡聚核苷酸探针进行Southern blot杂交、γ-32P检测及光密度扫描法测定端粒平均长度。结果各年龄段TRF值随着年龄的增长,其TRF值有下降的趋势,在20～35岁,36～55岁两年龄段之间无统计学差异,在35岁后其TRF值为6.64+0.21kb,在56岁后其TRF值为4.41+0.09kb,进行统计学分析后有统计学显著性差异。当FSH值小于30U/L时,其TRF值虽有下降趋势,无明显统计学差异,而FSH为31～40U/L时,其TRF值5.76±0.23kb,与前2组相比较有明显统计学差异;卵巢早衰组与正常绝组的TRF相比有明显统计学差异,卵巢早衰组女性的TRF值明显长于正常绝经组。结论对于正常女性来说,卵巢皮质TRF长度可以在一定程度上反映卵巢功能及生育能力,由于导致卵巢早衰的病因复杂及影响端粒长度的因素较多,导致端粒长度变化与卵巢功能相关性的临床意义尚未完全确定。     

自身免疫性甲状腺疾病患者外周血淋巴细胞端粒长度的研究

目的：研究自身免疫性甲状腺疾病（ATD）患者外周血淋巴细胞端粒长度的变化。方法：用流式原位杂交法检测38例ATD患者和48例健康对照者外周血淋巴细胞的端粒长度。结果：患者组外周血淋巴细胞端粒长度明显短于健康对照组（P＜0．001）,端粒长度与病程、发病时间、甲状腺激素水平不相关。健康对照组端粒长度随年龄增加而变短,病人组端粒长度与年龄不相关。结论：ATD病人的外周血淋巴细胞端粒长度比正常人短,且与年龄不相关,提示其外周血淋巴细胞复制、分裂增多及凋亡异常。     

端粒在心血管疾病研究中的意义

端粒是真核细胞染色体末端的特殊结构 ,其长度随细胞增殖不断缩短。近来通过对动脉粥样硬化 (AS)和高血压病的研究发现心血管系统的疾病也可出现组织细胞或外周血白细胞端粒长度的缩短。     

荧光定量PCR测定端粒长度

目的 应用实时荧光定量聚合酶链式反应(Q-PCR)方法测定端粒长度.方法 选取9种人类细胞株,提取基因组DNA,采用Q-PCR方法测定相对T/S比率,DNA印迹法测定末端限制性片段(TRF)长度,进行二者之间的相关性分析.结果 定量PCR测定端粒长度相对T/S比率为0.68±0.57,DNA印迹法测量平均TRF值为8.57±2.34,两种方法测定结果的相关性分析R2=0.7807(P＜0.01).结论 采用荧光定量PCR方法测量端粒长度具有重复性好、省时、简便、可靠的特点,可高通量处理大量样品.     

端粒在心血管疾病研究中的意义

端粒是真核细胞染色体末端的特殊结构，其长度随细胞增殖不断缩短。近来通过对动脉粥样硬化(AS)和高血压病的研究发现心血管系统的疾病也可出现组织细胞或外周血白细胞端粒长度的缩短。     

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.