果糖和二硫苏糖醇对骨髓间充质干细胞冻存后活性及多能性的影响

背景:冻存是保证干细胞临床应用的关键步骤之一,但现有冻存技术常导致细胞活性降低、多能性丧失及分化能力下降。目的:探究果糖及二硫苏糖醇是否有助于维持冻存后骨髓间充质干细胞多能性及成骨分化潜能。方法:分离培养SD大鼠骨髓间充质干细胞,在细胞冻存前分别用果糖(200μmol/L),二硫苏糖醇(500μmol/L)μmol/L)及果糖(200μmol/L)+二硫苏糖醇(500预处理1 h。冻存6个月后,复苏细胞并用倒置显微镜观察细胞形态,MTT实验检测细胞活性,定量PCR检测相关干性基因(Nanog,Oct4及Sox2)的表达,碱性磷酸酶活性测试及茜素红染色检测复苏骨髓间充质干细胞成骨分化能力。结果与结论:(1)复苏后各组细胞在形态上无明显差别;(2)果糖预处理及联合预处理有助于骨髓间充质干细胞活性维持;(3)二硫苏糖醇预处理可显著促进骨髓间充质干细胞多能性相关基因Nanog及Sox2的表达;(4)果糖、二硫苏糖醇及联合预处理皆有助于维持骨髓间充质干细胞成骨分化潜能,但以二硫苏糖醇及联合预处理组效果最佳;(5)结果表明,果糖预处理有助于维持冻存骨髓间充质干细胞活性,二硫苏糖醇有助于维持冻存骨髓间充质干细胞多能性及成骨分化能力。     

煅烧骨/壳聚糖复合材料促进SD大鼠骨髓间充质干细胞的成骨分化

背景:课题组前期研究已经证实煅烧骨/壳聚糖复合材料的微观结构与自然骨组织相似,安全无毒,抗压强度良好,并且能够促进成骨细胞的增殖及黏附,因此继续研究其对骨髓间充质干细胞成骨分化的作用具有重要意义。目的:探讨煅烧骨/壳聚糖复合材料对骨髓间充质干细胞成骨分化的影响。方法:将煅烧骨/壳聚糖复合材料与SD大鼠骨髓间充质干细胞体外共培养并进行成骨诱导,成骨诱导1,2,3周时,采用实时荧光定量PCR检测成骨分化相关基因的表达。结果与结论:煅烧骨/壳聚糖复合材料能促进骨髓间充质干细胞的增殖黏附,并促进成骨相关基因碱性磷酸酶、Ⅰ型胶原、骨钙素的表达,具有诱导骨髓间充质干细胞成骨分化的作用。     

C57小鼠脂肪及骨髓间充质干细胞生物学特性的比较

背景：研究发现,脂肪源干细胞具有和骨髓间充质干细胞一样的贴壁和形成成纤维样克隆特性,并具有向骨、脂肪、软骨等多系分化的能力。 目的：比较C57小鼠脂肪源间充质干细胞和骨髓间充质干细胞的生物学特点。 方法：在无菌的条件下分别从C57小鼠的脂肪和骨髓中获取脂肪间充质干细胞和骨髓间充质干细胞。体外分离、培养并将脂肪间充质干细胞和骨髓间充质干细胞传至第3代,进行细胞形态、表面标记、生长动力学分化潜能测定和Notch信号相关基因的检测。 结果与结论：脂肪间充质干细胞和骨髓间充质干细胞形态学相似,第3代的脂肪间充质干细胞和骨髓间充质干细胞均表达CD29、CD105、Sca-1,不表达CD34、CD133,但骨髓间充质干细胞还表达CD45;生长曲线和细胞克隆分析显示脂肪间充质干细胞的增殖速度明显比骨髓间充质干细胞快;脂肪间充质干细胞和骨髓间充质干细胞均可向成骨、成脂、成软骨诱导分化,脂肪间充质干细胞更易向成骨诱导;Notch相关基因检测显示脂肪源干细胞的Jagged-1表达水平明显比骨髓间充质干细胞低,而Hes-1的表达水平脂肪源干细胞明显高于骨髓间充质干细胞的表达水平。提示脂肪间充质干细胞比骨髓间充质干细胞扩增能力更强,更易向成骨分化,可能与Hes-1表达水平有关。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

胎儿真皮间充质干细胞中的全能性相关转录因子的表达

背景：人真皮纤维母细胞或间充质干细胞可形成诱导性多能干细胞,但不同研究者所用的转录因子组合却并不相同。 目的：分离人胎儿真皮间充质干细胞,检测其全能性相关转录因子的表达。 方法：水囊引产5月龄胎儿,按照既往分离培养间充质干细胞的方法,得到胎儿真皮间充质干细胞。 结果与结论：胎儿真皮间充质干细胞高表达Oct4和C-myc、中度表达Sox2,是形成诱导性多能干细胞较好的体细胞。     

敲降ZEB1基因表达对人脂肪间充质干细胞增殖、周期与凋亡的影响

目的探讨敲降ZEB1基因表达对人脂肪间充质干细胞(h ADSCs)增殖、周期和凋亡的影响。方法胶原酶消化法提取人脂肪组织中原代间充质干细胞,对其进行免疫学表型、成骨和成脂分化能力鉴定后用于实验。脂质体转染法将siRNA转入h ADSCs,实时定量PCR和Western blot检测ZEB1、细胞增殖、周期和凋亡相关基因mRNA和蛋白的表达。MTS法检测细胞增殖。流式细胞术检测细胞周期和细胞凋亡率。结果与si-NC转染组相比,si-ZEB1转染组可使ZEB1 mRNA和蛋白表达水平降低(P0.01),细胞增殖能力明显下降(P0.05),增殖相关基因CCND1、MKI67、MYC和PCNA表达显著下调(P0.05或P0.01);G1期细胞比例从50.71%增加到58.94%(P0.01),S期和G2期细胞比例分别减少了6.16%和2.07%;细胞凋亡率上升了10.2%,促凋亡相关基因TP53和BAX表达上调,抗凋亡基因MCL1和BCL2表达下调(P0.05或P0.01)。结论 ZEB1具有促进h ADSCs增殖和周期进展,抑制细胞凋亡的作用。     

人脐带间充质干细胞的生物学性质研究进展

人脐带中富含间充质干细胞(MSCs)。这些细胞能表达多种间充质干细胞标志物及多种干细胞相关基因,能分化为3个胚层衍生的多种成熟细胞,合成多种营养因子和细胞因子,支持造血干细胞等细胞的增殖和功能,并具有低免疫原性。     

Twist1、SIRT1、FGF2、TGF-β3基因在胎盘、脐带和乳牙三种间充质干细胞的表达及增殖调控作用研究

目的比较细胞增殖相关基因Twist1、SIRT1、FGF2、TGF-β3在胎盘、脐带和乳牙3种间充质干细胞中的表达差异,分析其对3种间充质干细胞的增殖调控作用。方法通过显微镜对不同传代次数的3种间充质干细胞的形态结构进行比较;经过体外传代培养,利用荧光定量PCR测定细胞增殖相关基因Twist1、SIRT1、FGF2、TGF-β3的表达水平;采用MTT法检测并比较分析3种间充质干细胞的增殖及促增殖能力。结果Twist1、TGF-β3基因在胎盘间充质干细胞中表达水平最高,FGF2基因的表达水平最低,SIRT1基因在脐带间充质干细胞中的表达水平较高。随着传代培养进行,4种基因表达水平在不同代次的间充质干细胞中的表达水平各不相同。Twist1、SIRT1、TGF-β3表达上调时3种间充质干细胞增殖能力增强,FGF2表达上调时3种间充质干细胞增殖能力减弱。结论Twist1、SIRT1、FGF2、TGF-β3在3种间充质干细胞中的表达水平存在一定差异;Twist1、SIRT1、TGF-β3对3种间充质干细胞的增殖表现为正调控作用;而FGF2基因则相反表现为负调控作用。     

沉默膜联蛋白A1基因影响兔骨髓间充质干细胞的增殖及成骨分化能力*☆

背景：膜联蛋白A1蛋白参与细胞的增殖和分化的调控。激素性股骨头坏死与骨髓间充质干细胞增殖和分化能力的改变相关。 目的：观察沉默膜联蛋白A1基因对兔骨髓间充质干细胞增殖及成骨分化能力的影响。 方法：将新西兰大白兔的骨髓间充质干细胞分离培养后分为RNA干扰组,用携带针对膜联蛋白A1基因的短发夹RNA慢病毒载体LV-shANXA1感染骨髓间充质干细胞下调膜联蛋白A1基因的表达;阴性对照组,用携带无义基因序列的慢病毒载体LV-shANXA1-NC感染骨髓间充质干细胞;空白对照组,不加任何干预措施。 结果与结论：纯化后的骨髓间充质干细胞CD44和CD105表达阳性率分别为97.2%和95.8%,而CD45阳性率为2.5%。经嘌呤霉素筛选后,感染复数为50时,LV-shANXA1对骨髓间充质干细胞感染效率为80%;感染复数为100时,感染效率为95%。实时 PCR 和Western blot检测显示感染复数为50时,LV-shANXA1对膜联蛋白A1基因的沉默效率效率可达72.4%以上。膜联蛋白A1表达下调后,骨髓间充质干细胞出现明显生长抑制,在随后成骨分化诱导后,细胞的碱性磷酸酶染色阳性率和碱性磷酸酶活性明显降低,且茜素红染色呈阴性反应。表明沉默膜联蛋白A1基因降低骨髓间充质干细胞增殖及成骨分化能力,这可能是激素性股骨头坏死发病机制之一。关键词：膜联蛋白A1;基因沉默;骨髓间充质干细胞;成骨分化;细胞增殖;激素性股骨头缺血性坏死 doi:10.3969/j.issn.1673-8225.2012.19.001     

低氧和氧化应激对不同来源间充质干细胞旁分泌的差异性影响

背景：间充质干细胞的生物学行为受所处生存微环境的影响，微环境条件预处理是调控间充质干细胞功能的重要手段。目的：对比氧化应激和低氧条件下脐带间充质干细胞、脂肪间充质干细胞旁分泌功能的差异，为治疗不同疾病选择适当的间充质干细胞预处理方式提供理论依据。方法：培养脐带间充质干细胞和脂肪间充质干细胞，分别加入不同浓度的H₂O₂或不同体积分数的O₂干预，检测细胞形态、增殖、活力和旁分泌因子表达。结果与结论：(1)普通培养环境下，脐带间充质干细胞中脑源性神经营养因子和转化生长因子β的表达水平显著高于脂肪间充质干细胞，而脂肪间充质干细胞中基质细胞衍生因子1α和肿瘤坏死因子刺激因子6的表达水平显著高于脐带间充质干细胞；(2)中低浓度水平(≤100μmol/L)H₂O₂对2种间充质干细胞活力的影响无显著差异，但H₂O₂浓度从50μmol/L增至100μmol/L使脐带间充质干细胞中血管内皮生长因子表达水平显著增高，脂肪间充质干细胞中碱性成纤维细胞生长因子...     

抑制外泌体释放对骨髓间充质干细胞生物学特性的影响

目的:探讨抑制骨髓间充质干细胞外泌体的释放对其生物学特性的影响及其机制。方法:利用TALEN技术靶向性敲除Rab27a基因构建外泌体释放缺失小鼠模型,随后分离、培养并鉴定骨髓间充质干细胞,用外泌体提取试剂盒提取培养基中的外泌体,并用纳米颗粒跟踪分析(NTA)计数外泌体的数量,透射电镜观察外泌体的大小及形态。利用Ed U实验及检测增殖细胞核抗原(PCNA)的表达来评价间充质干细胞的增殖能力。通过TUNEL染色及MTS实验检测间充质干细胞对缺氧的耐受能力。结果:敲除Rab27a的间充质干细胞释放外泌体的数量明显减少,抑制间充质干细胞外泌体的释放能显著降低其增殖及对缺氧的耐受能力。结论:抑制间充质干细胞外泌体的释放可导致其增殖能力及对缺氧的耐受能力明显降低。     

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.