bFGF对人关节软骨细胞增殖和凋亡的影响

目的：观察碱性成纤维细胞生长因子(bFGF)对体外单层培养的人负重关节软骨细胞的增殖和凋亡的影响。方法：对8例来自人负重关节软骨细胞系进行体外单层传代培养，从第2代起试验组培养液中加入bFGF，观察细胞的倍增时间(DT)、流式细胞分析细胞增殖周期和凋亡指数，并与无bFGF组相比较。结果：与未加bFGF相比较，加用10μg／ml bFGF后，DT缩短，S期细胞数升高，细胞凋亡无变化。结论：bFGF能促进培养的人关节软骨细胞生长，不引起细胞凋亡。     

大鼠肋生长板软骨细胞的分离、培养及生物学特性的研究

目的：建立大鼠肋生长板软骨细胞（RGC）分离、培养的方法，探讨其生物学特性，为软骨细胞增殖分化的调控和软骨组织工程修复的研究建立实验基础。 方法： 显微解剖出大鼠肋生长板软骨，酶消化法获得分散的单个RGC。观察单层培养的不同代数RGC的细胞形态变化,并进行细胞生长动力学分析；组织化学和免疫细胞化学方法检测不同代数RGC蛋白多糖和Ⅱ型胶原的表达。 结果： 本实验分离的RGC中活细胞比例大于98%；原代培养细胞呈多角形或圆形，第6代细胞仍保持多角的形态；前4代细胞的每日倍增指数随着传代次数的增加而增加，第5代后显著降低；原代培养的RGC中Ⅱ型胶原阳性染色细胞比例大于95%，阿尔新蓝染色也呈阳性；随着传代次数增加阳性细胞的比例下降。 结论： 本研究所建立的分离培养方法可以获得高纯度、高活性的软骨细胞，前3代RGC保持了在体软骨细胞的表型是研究软骨增殖分化调控的良好材料。     

自体血清与胎牛血清培养兔关节软骨细胞的生物学特性

背景：目前培养软骨细胞多使用胎牛血清。但近年异种血清培养组织向临床应用的安全性受到质疑,因此自体血清培养越来越受到重视。 目的：比较体积分数10%兔自体血清与体积分数10%胎牛血清培养兔关节软骨细胞生物学特性的差异。 方法：兔自体血清培养液制备后,分离培养兔关节软骨细胞,分别在体积分数10%自体血清和体积分数10%胎牛血清中进行单层传代培养至1,3,5代,采用光镜观察细胞形态变化,绘制生长曲线评估细胞增殖速度,观察甲苯胺蓝染色、Ⅰ,Ⅱ型胶原免疫组织化学染色结果,以流式细胞仪分析细胞Ⅰ,Ⅱ型胶原以及CD26,CD44的表达变化。 结果与结论：①在细胞形态上自体血清和胎牛血清培养的软骨细胞差异不大。②自体血清培养的软骨细胞较胎牛血清培养的软骨细胞生长速度更快。③甲苯胺蓝染色结果示,无论是自体血清还是胎牛血清所培养的细胞,染色随代龄的增加逐渐变浅,细胞传至第5代时两组几乎均无异染。对于1代和3代细胞而言,自体血清培养的软骨细胞较胎牛血清所培养的软骨细胞较为深染。④Ⅰ型胶原的表达随代数的增加而增加,而Ⅱ型胶原的表达则随代数的增加而减少。在3代时自体血清培养的软骨细胞Ⅰ型胶原的表达水平低于胎牛血清培养的软骨细胞(P < 0.05);在1代和3代时自体血清培养的软骨细胞Ⅱ型胶原的表达水平高于胎牛血清培养的软骨细胞(P < 0.05)。⑤CD26表达呈现先升高后降低的趋势,而CD44的表达不随传代数的增加而改变。提示同异体体积分数10%的胎牛血清相比,体积分数10%的自体血清培养的兔软骨细胞生长速度快,且在大部分指标上可以较好地保持细胞表型。     

Ⅱ型胶原酶消化法培养兔关节软骨细胞

背景：目前关节软骨细胞的分离培养技术已经比较成熟,但是研究中发现通过目前技术培养的软骨细胞生长周期慢,容易出现退变现象,不利于后续试验的进行。 目的：改进并探讨4周龄新西兰大白兔膝关节软骨细胞的分离与培养的方法。 方法：无菌条件下取4周龄新西兰大白兔双侧膝关节软骨,采用Ⅱ型胶原酶消化并机械吹打的方法,分离关节软骨细胞并进行原代、传代培养;采用形态学观察,甲苯胺蓝染色以及Ⅱ型胶原免疫组织化学方法对关节软骨细胞进行鉴定;MTT法检测关节软骨细胞增殖情况。 结果与结论：倒置显微镜下见膝关节软骨分离的原代软骨细胞6 h后开始贴壁,72 h可形成单层,96 h即可传代;前3代软骨细胞表型稳定,增殖力良好;第4,5代软骨细胞增殖能力减弱,绝大部分细胞变为长梭形和不规则形状。甲苯胺蓝染色显示培养的软骨细胞细胞质染成浅蓝色,细胞核染成深蓝色;免疫组织化学显示软骨细胞Ⅱ型胶原呈黄褐色阳性表达;MTT法检测表明前3代软骨细胞增殖差异无显著性意义(P > 0.05),且第1-3代与4代软骨细胞在培养第4-7天时,吸光度值差异有显著性意义(P < 0.05),与第5代软骨细胞在培养1-7天时,吸光度值差异有显著性意义(P < 0.05)。提示采用Ⅱ型胶原酶消化并机械吹打的方法能够获得大量生长速度快且不宜退变的新西兰大白兔膝关节软骨细胞,且培养的前3代新西兰兔膝关节软骨细胞最为适宜。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

碱性成纤维细胞生长因子对体外培养的乳兔软骨细胞中p53 mRNA表达的影响

目的 探讨在乳兔关节软骨细胞体外培养过程中,成纤维细胞生长因子（bFGF）对p53mRNA表达水平的影响及意义。方法 原代体外培养乳兔关节软骨细胞并传代,实验组加入10ng／ml的bFGF,光镜下观测各代细胞形态学变化,RT—PCR检测各代细胞中p53基因在mRNA水平上的表达。结果光镜观察体外培养的软骨细胞,对照组第4代培养细胞始出现凋亡细胞,bFGF组第7代出现少量凋亡细胞;RT—PCR检测bFGF组p53的目的基因指数（p53吸光光度值,β—actin吸光光度值）明显低于对照组（P〈0．05）。结论 在体外培养的兔关节软骨细胞中bFGF下调p53基因mRNA水平表达。     

体外诱导人骨髓间充质干细胞分化为软骨细胞的研究

目的：探讨胰岛素样生长因子-Ⅰ（IGF-Ⅰ）和软骨细胞培养液体外诱导骨髓间充质干细胞（MSCs）向软骨细胞分化的可能性。方法：采用Percoll分离液分离培养人胚MSCs，体外扩增，用流式细胞仪测定MSCs的CD44、CD71、CD34、CD45表达；在第4代MSCs中加入的条件培养基进行诱导，根据条件培养基的不同分为4组：①IGF-Ⅰ组：培养基中加入100ng／mL IGF-Ⅰ；②软骨细胞培养液组：加入软骨细胞培养液；③联合诱导组：培养液中加入100ng／mL IGF-Ⅰ和软骨细胞培养液；④对照组：培养基中不加入IGF-Ⅰ和软骨细胞培养液。采用倒置显微镜观察、Ⅱ型胶原免疫组化、细胞内蛋白多糖（PG）含量测定等方法判断诱导细胞的形态变化和表达软骨基质的能力。结果：体外培养的MSCs呈现成纤维细胞样形态，流式细胞仪检测结果显示，MSCs阳性表达CD44，阴性表达CD34，CD45；IGF-Ⅰ和软骨细胞培养液联合诱导15d后，MSCs开始呈现软骨细胞的形态特点，Ⅱ型胶原免疫组化呈阳性，而未诱导的MSCs组呈阴性；联合诱导组细胞内PG含量为8．92μg／ml，显著高于IGF-Ⅰ组、软骨细胞培养液组和对照组，但低于正常软骨细胞（P〈0．01）。结论：IGF-Ⅰ和软骨细胞培养液能诱导MSCs向软骨细胞分化。     

大骨节病关节软骨胶原表型表达和软骨细胞异常分化的研究

目的探讨大骨节病关节软骨胶原表型表达的变化特点和软骨细胞异常分化在发病中的意义。方法用单克隆免疫组化法测定５例大骨节病关节软骨Ⅰ、Ⅱ、Ⅲ、Ⅵ、Ⅹ型胶原表型的表达。结果（１）关节软骨表层的Ⅱ型胶原表型表达减少；（２）Ⅰ、Ⅲ和Ⅵ型胶原表型表达见于关节软骨全层，而Ⅹ型胶原仅限于关节软骨钙化层和深层软骨细胞团周围；（３）软骨细胞团有Ⅰ、Ⅱ、Ⅲ、Ⅵ型胶原表型表达，而软骨细胞坏死区内无任何胶原表型表达。结论大骨节病关节软骨胶原表型表达类似于原发性骨关节病的变化，但在关节软骨表层Ⅰ型胶原表型表达增强以及软骨坏死区内无任何胶原表型表达不同于原发性骨关节病。     

Ⅱ型胶原酶消化法可短时间获得大量纯化大鼠关节软骨细胞

背景：建立一种经济、快捷、切实可行的软骨细胞分离培养体系对于软骨体外实验研究有着重要的意义。 目的：探讨与改进大鼠关节软骨细胞的培养方法。 方法：无菌条件下取新生1周龄SD雄性大鼠双侧髋及膝关节软骨,采用Ⅱ型胶原酶消化法分离软骨细胞并进行原代、传代培养及鉴定。 结果与结论：倒置相差显微镜下见原代培养的软骨细胞12 h后开始贴壁,3 d左右可形成单层,4 d左右即可传代。传至第6代后,部分细胞变为梭形;第7代后,绝大部分细胞变为长梭形和不规则形状,增殖能力减弱。甲苯胺蓝染色显示培养的软骨细胞核呈异染性,免疫荧光染色显示培养的软骨细胞Ⅱ型胶原呈阳性表达。说明采用此方法可在短时间内获得大量纯化的大鼠软骨细胞。     

体外传代培养兔关节软骨细胞的去分化现象

背景：兔是软骨组织工程研究中应用非常广泛的实验动物模型,关节软骨细胞的去分化现象已经被广泛认可。 目的：观察体外传代培养过程中兔关节软骨细胞的去分化现象。 方法：将从新西兰大白兔膝关节分离获取的原代软骨细胞进行传代培养至第7代,对细胞生长、形态、基质分泌以及基因表达等方面分别采用细胞计数,显微镜观察,F机动蛋白染色、番红O染色,糖胺聚糖定量测定以及半定量聚合链式反应进行鉴定和比较。 结果与结论：光学显微镜下,兔关节软骨细胞在体外传代培养过程中细胞形态由小且圆形或多角形,逐步转变大且为成纤维细胞样的梭形形态,对F机动蛋白的染色进一步佐证了这样的形态变化。细胞计数结果表明,细胞增殖能力随代次增加显著下降,特别是第3代以后的软骨细胞基本无明显增殖;经过番红O染色以及定量测定糖胺聚糖的含量,发现软骨特性胞外基质分泌量从第2代细胞开始就呈现显著的降低。半定量聚合链式反应检测结果表明,随着传代次数的增加,特别是第3代以后,软骨相关特征分子(包括Ⅱ型胶原、聚集蛋白聚糖、软骨寡聚基质蛋白和SOX9等)基因表达水平下调;而与去分化相关的特征分子Ⅰ型胶原和多能蛋白聚糖基因表达水平上调,同时,细胞表面分子CD90基因表达上调,而CD14基因表达未见明显变化。结果证实,兔软骨细胞在体外传代过程中可出现快速地去分化现象,呈现出特征基因表达水平的变化,第3代以内的软骨细胞适合应用于软骨组织再生修复。     

自体血清培养新西兰大白兔关节软骨细胞

目的：改进与探讨新西兰大白兔软骨细胞体外培养的方法。方法：取3月龄新西兰大白兔自体血,制备自体血清备用,无菌条件下取4周龄新西兰大白兔双侧膝关节软骨,采用Ⅱ型胶原酶消化并机械吹打的方法,分离关节软骨细胞并应用新西兰大白兔自体血清进行原代、传代培养;采用形态学观察,甲苯胺蓝染色以及Ⅱ型胶原免疫组织化学方法对膝关节软骨细胞进行鉴定,MTT法检测软骨细胞的增值能力;结果：倒置显微镜下见原代软骨细胞2 h后开始贴壁,8 h可形成单层,24 h即可传代。第1~5代细胞表型稳定,增殖力良好。甲苯胺蓝染色显示培养的软骨细胞核呈深染色,细胞质呈淡蓝色;免疫组织化学显示软骨细胞Ⅱ型胶原呈黄褐色表达,MTT检测显示前5代软骨细胞增值能力强,无明显差异;结论：结果表明自体血培养的前5代新西兰大白兔节软骨细胞均可用于膝骨关节炎的研究。     

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.     

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.     

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 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.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.