长链非编码MALAT-1 调控miR-205 的表达影响非小细胞肺癌细胞的生长和迁移

目的:探讨在非小细胞肺癌中,LncRNA MALAT-1 与miR-205 的相互关系,以及影响肺癌细胞生物学行为的机制。方法:qPCR 检测不同非小细胞肺癌中LncRNA MALAT-1 的表达情况;双荧光素酶报告基因检测MALAT-1 与miR-205的相互作用;Transwell 侵袭实验和划痕实验检测抑制MALAT-1 后肺癌细胞侵袭能力的变化,以及抑制miR-205 的表达后肺癌细胞迁移和侵袭能力的恢复情况;裸鼠皮下成瘤检测抑制LncRNA MALAT-1 后肺癌细胞体外成瘤体积和质量变化。结果:与其他肺癌细胞株相比,A549 细胞中MALAT-1 表达最高,miR-205 的表达水平最低;双荧光素酶实验证实MALAT-1 能与miR-205 的3忆UTR 特异性结合,可以调控miR-205 的表达与活性;抑制MALAT-1 的表达后可以降低肺癌细胞的迁移和侵袭能力;抑制miR-205 的表达水平过后,肺癌细胞的迁移和侵袭能力相对增强;抑制MALAT-1 的表达后,荷瘤小鼠的肿瘤体积和重量都明显减小。结论:MALAT-1 可以调控miR-205 的表达影响肺癌细胞A549 的侵袭和迁移能力。     

肺腺癌转移相关转录因子1在口腔鳞状细胞癌组织中的表达及临床意义

目的分析口腔鳞状细胞癌组织中长链非编码RNA(lncRNA)肺腺癌转移相关转录因子1(MALAT1)的表达及其临床意义。方法收集口腔鳞癌组织标本81例和正常口腔黏膜组织20例,分别设为口腔鳞癌组和健康对照组。采用RT-PCR方法检测2组组织中lncRNA MALAT1表达情况,统计分析lncRNA MALAT1表达与病理资料特征的关系,Kaplan-Meier分析lncRNA MALAT1表达和生存预后间的关系。结果与健康对照组相比,lncRNA MALAT1在口腔鳞癌组中表达明显增高,差异有统计学意义(P 0. 05);口腔癌组织中lncRNA MALAT1的相对表达量在不同肿瘤分期、肿瘤分化程度和有无颈部淋巴结转移间比较,差异均有统计学意义(P 0. 05);而在不同年龄、性别、肿瘤大小及肿瘤类型间比较,差异均无统计学意义(P 0. 05); lncRNA MALAT1高表达组和低表达组患者的3年总体生存率(OS)分别为45. 9%(17/37)、65. 9%(29/44),Kaplan-Meier生存分析(log-rank检验)表明MALAT1高表达组患者3年OS显著低于MALAT1低表达组患者,差异有统计学意义(P 0. 05)。结论口腔鳞癌组织中lncRNA MALAT1表达增高,其表达水平与口腔鳞癌患者的不良预后有关,在口腔鳞癌组织中检测其表达可能有助于肿瘤的诊断及作为药物治疗的靶点。     

长链非编码RNA MALAT1靶向miR-129对前列腺癌PC3细胞增殖和侵袭的影响

目的 探讨长链非编码RNA MALAT1对前列腺癌PC3细胞增殖和侵袭的影响,初步分析其机制.方法 将lncRNA MALAT1抑制物转染前列腺癌PC3细胞,分为untreated组(对照组)、si-NC组(阴性对照组)及si-MALAT1组(沉默MALAT1),实时定量PCR检测各组PC3细胞lncRNA MALAT1和miR-129的表达,双荧光素酶报告基因实验分析lncRNA MALAT1和miR-129的靶向关系,CCK-8方法检测各组PC3细胞增殖能力,Transwell实验检测各组PC3细胞侵袭能力.结果 转染MALAT1抑制物能够降低PC3细胞lncRNA MALAT1的表达,上调miR-129的表达(P＜0.05);双荧光素酶实验表明lncRNA MALAT1可以靶向调控miR-129.lncRNA MALAT1表达抑制后,PC3细胞的增殖与侵袭能力下降(P＜0.05).结论 沉默lncRNA MALAT1可以抑制前列腺癌细胞PC3的增殖和侵袭,其机制可能与调控miR-129表达有关.     

非小细胞肺癌中微小卫星体不稳定和其它分子学异常

微小卫星体是高度多形的散在于基因组内短的前后排列的核苷酸重复序列。微小卫星体不稳定被认为是肿瘤发生的一个新的分子机制,它反映了误配修复基因包括hMSH_2和hMLH_1基因异常的多种复制错误。除遗传性非息肉型结直肠癌(HNPCC)和某些散发性的肿瘤外,小细胞肺癌(SCLC)也发现有45％的微小卫星体不稳定。为了研究非小细胞肺癌(NSCLC)中微小卫星体不稳定,作者用多个微小卫星体包括4个双核苷酸、1个四核苷酸和1个     

新型m6A阅读器IGF2BP1在非小细胞肺癌组织中的表达及临床意义

目的本研究拟分析新型m6A阅读器-胰岛素样生长因子2(IGF2)mRNA结合蛋白1(IGF2BP1)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达水平、临床意义及对人非小细胞肺癌细胞增殖水平的影响。方法利用癌症基因组图谱(TCGA)数据库下载NSCLC相关数据集,分析肿瘤组织与正常组织之间IGF2BP1表达的差异;利用Kaplan-Meier曲线进行患者生存分析;MTT方法检测细胞增殖水平。结果IGF2BP1蛋白主要表达于肿瘤细胞的胞浆和细胞膜。TCGA分析发现与癌旁组织比较,IGF2BP1在NSCLC组织中的表达明显增加(P<0.05);Kaplan-Meier分析发现,IGF2BP1表达与非小细胞肺癌患者OS时间相关,IGF2BP1高表达的非小细胞肺癌患者,OS时间明显缩短(P<0.05)。转染IGF2BP1 siRNA至非小细胞肺癌细胞后发现,与阴性质粒转染组比较:转染siRNA-IGF2BP1的非小细胞肺癌细胞,IGF2BP1表达明显降低;IGF2BP1表达下调后细胞增殖水平减慢。结论IGF2BP1是一个新的NSCLC生物标记物,可作为潜在的治疗新靶点。     

LncRNA MALAT-1 对弥漫性大B 细胞淋巴瘤增殖和耐药性的影响

目的:探讨长链非编码RNA(LncRNA)MALAT-1对弥漫性大B细胞淋巴瘤(DLBCL)增殖和耐药性的影响。方法:收集2015年3月至2018年11月我院81例DLBCL患者的淋巴组织标本作为研究对象,另选取60例健康淋巴组织标本作为对照组。采用实时荧光定量PCR检测MALAT-1在DLBCL组织和健康淋巴组织中的mRNA表达水平;构建NC-shRNA和MALAT-1-shRNA慢病毒稳定细胞系,CCK-8法检测MALAT-1下调对DLBCL细胞增殖的影响;在耐药性分析中,NC-shRNA和MALAT-1-shRNA细胞用200 ng/ml阿霉素分别处理48 h。采用Western blot检测NC-shRNA和MALAT-1-shRNA细胞耐药基因MDR1和MRP1的蛋白表达水平。结果:DLBCL组织中MALAT-1的mRNA相对表达水平显著高于健康淋巴组织,差异有统计学意义(1.46±0.11 vs 0.41±0.08,P0.05);CCK-8结果显示,与NC-shRNA组相比,MALAT-1-shRNA组细胞在24 h、36 h、48 h、72 h时的增殖能力均显著下降,差异有统计学意义(P0.05);耐药性检测结果表明,与NC-shRNA组、MALAT-1-shRNA组和NC-shRNA+Adriamycin组比较,MALAT-1-shRNA+Adriamycin组增殖曲线的平均斜率下降,差异有统计学意义(P0.05)。Western blot结果显示,与NC-shRNA组比较,MALAT-1-shRNA组细胞中MDR1和MRP1的蛋白表达水平显著下调(P0.05)。结论:LncRNA MALAT-1在DLBCL组织中呈高表达,下调MALAT-1的表达可显著抑制DLBCL细胞的增殖能力和耐药性。     

非编码基因MALAT1功能性序列真核表达载体的构建及其在SW620细胞中的表达

目的构建人肺腺癌转移相关转录本1（MALAT1）功能性序列重组真核荧光表达载体,并在人大肠癌SW620细胞株中表达。方法采用RT-touchdown PCR方法,从人正常大肠黏膜组织中提取总RNA,扩增MALAT1功能性序列,将扩增产物克隆至pTA2载体,测序证实后,克隆至pEGFP-C1质粒中,构建重组真核荧光表达载体;阳离子脂质体介导下转染人大肠癌SW620细胞,荧光显微镜检测细胞中增强型绿色荧光蛋白（EGFP）的表达。通过Real-time PCR检测质粒转染前后MALAT1/fgf mRNA表达水平的变化。结果经pTA2载体和pEGFP-C1载体克隆、酶切鉴定及序列分析后,证实人MALAT1功能性序列重组真核荧光表达载体构建成功。在荧光显微下观察到带绿色荧光的SW620细胞。pEGFP-C1-MALAT1/fgf转染前后SW620细胞MALAT1/fgf mRNA表达水平倍比关系为2.376±0.573,差异具有统计学意义（P〈0.05）。结论成功构建了人MALAT1功能性序列真核荧光表达载体pEGFP-C1-MALAT1/fgf,转染SW620细胞后可引起MALAT1/fgf mRNA表达上调,为进一步研究MALAT1的结构、功能及其与人大肠癌的关联奠定了基础。     

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.     

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.     

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.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.