体外诱导绵羊红细胞特异性抗体形成及单纯疱疹病毒感染对它的影响

本文建立了一种较稳定、理想的人淋巴细胞体外诱导绵羊红细胞(SRBC)特异性抗体生成的系统。用SRBC体外刺激人扁桃体淋巴细胞,用溶血空斑法计数针对SRBC特异性抗体形成细胞。发现极低量抗原可诱导其抗体形成,抗体形成量随抗原量呈规律性变化;在抗原刺激后的第4天特异性抗体开始出现,第6天达高峰,并稳定维持至第8天;在辅助刺激剂美洲商陆(PWM)存在下,抗体形成量显著高于无PWM的情况;除去人扁桃体细胞中粘附细胞(主要是巨噬细胞)才能诱导最适抗体形成。将具感染性的HSV-1与SRBC一起加入淋巴细胞培养中,可显著抑制SRBC诱导的特异性抗体形成,这一抑制效应与病毒的感染量有关。此系统中同时加入α-干扰素则可部分解除病毒的抑制效应,并且解除效果与α-干扰素的剂量有关。     

用Epstein-Barr病毒转化法获得分泌肾综合征出血热病毒抗体的B淋巴细胞系

人B淋巴细胞膜上带有Epstein-Barr(EB)病毒受体,用EB病毒转化人B淋巴细胞是获得人源性单克隆抗体的重要方法之一。最近David等报道,先用EB病毒转化感染巨细胞病毒(CMV)患者的脾脏B淋巴细胞,再将转化了的B淋巴细胞和人骨髓瘤细胞WL-L_2-727融合,获得迄今为止分泌抗体滴度最高的人—人杂交瘤312A和914,能稳定分泌抗体达12     

用体外免疫的人淋巴细胞建立抗破伤风类毒素的杂交瘤克隆

我们应用破伤风类毒素体外免疫的人扁桃体细胞和人-鼠异源骨髓瘤RF 系细胞进行融合,从中筛选到一株杂交瘤细胞89112—50,并通过克隆化筛选获得了两个亚克隆,其分泌的抗体是抗原特异的,和三种抗原(OVA、TC-S、FYG)都不交叉,分泌抗体的功能也比较稳定,在培养瓶内连续扩增传代13次后,仍维持相当高的抗体分泌能力,在常规传代培养过程中所收集的培养物上清液中抗体的含量平均为69.6μg/ml。     

EPSTEIN-BARR病毒血清抗体阳性健康人外周血体外培养“自发性”转化的研究

对35名Epstein-Barr病毒血清抗体阳性健康人的外周血淋巴细胞进行了体外培养,观察B淋巴细胞感染了EB病毒后所导致“自发性”转化的情况。由于在培养基中加入了免疫抑制剂环胞菌素A,使“自发性”转化的发生率由26.7％提高到74.3％,说明了在血清抗体阳性健康人的外周血液中,EB病毒感染的B淋巴细胞的数量远比既往文献报道的高。     

人NK细胞克隆化培养条件的初步探讨

为探讨NK细胞克隆化培养的条件,我们首先将人外周血单个核细胞经rhIL-2诱导,使NK细胞得以扩增后,去除T细胞,得到相对纯化的NK细胞.经有限稀释,在饲养细胞及rhIL-2、PHA及LCM等培养条件下,获得NK细胞的单个克隆并进行鉴定.结果表明,每96孔板可获4-16个CD3-CD56+NK克隆,每个克隆的细胞数最多可达2.35×104,存活约3-5周.以丝裂霉素C(25μg/m1)作为饲养细胞抑制剂,以rhIL-2 200u/ml、PHA 10μg/ml及10%LCM培养,可获得较多的克隆和细胞数.     

流行性出血热病毒(EHFV)在传代人淋巴细胞上的致病变作用增殖的研究

流行性出血热病毒陈株、L_99和SR_11株按种传代淋巴细胞克隆株Ly-A_6后第5～7日观察到CPE,开始时受感染细胞浆内颗粒增多,细胞圆缩、聚积成堆,随后细胞膜模糊不清,2～3天脱落,同时制片IFA检测到胞浆内特异性荧光颗粒,以上结果能被EHF免疫血清所中和。接种病毒后的细胞单层用l％甲基纤维素加维持液覆盖,第6日吸去覆盖液,加5％甲醛-1％结晶紫固定染色可见到明显的PFU。EHFV致病变细胞株的染得将为病原学研究提供了优越的基质细胞。     

一种适用于骨髓瘤细胞、淋巴细胞杂交瘤细胞生长的无血清培养液CBSF

本文报道CBSF是一种运用于骨髓瘤细胞、淋巴细胞杂交瘤细胞的无血清培养液。我们成功地培养了几株骨髓瘤和杂交瘤细胞,它们能长期在其中生长繁殖传代,并保持杂交瘤细胞能持续分泌抗原特异的抗体。     

玉米赤霉烯酮单克隆抗体和免疫酶技术研究

采用蛋白质连接技术合成玉米赤霉烯酮抗原,免疫Balb/c鼠,通过淋巴细胞杂交瘤技术建立六株分泌抗玉米赤霉烯酮的单克隆抗体杂交瘤细胞株。间接酶联免疫吸附试验测定细胞上清抗体效价为1:2084(4H8)、1:256(6H9、4H3、2H5、2C8)、1:16(3F10);腹水抗体效价为10~9(4H3、4H8)、10~8(2H5)、10~7(6H9)、10~5(3H10)。竞争间接酶联免疫吸附试验测定六株单克隆抗体对玉米赤霉烯酮的敏感度为0.3—0.8ng/ml。六株抗体与玉米亦霉烯醇的交叉反应率为1.3—9.0％。六株单克隆抗体均属IgG类。细胞体外传代培养和冻存复苏后分泌抗体稳定。纯化抗体在37℃保存12天稳定,-30℃保存90天抗体滴度不变。用该抗体建立竞争间接酶联免疫吸附试验检测掺合玉米赤霉烯酮的玉米、小麦、饲料,平均回收率分别为105％、90％、103％,平均批间变异系数为5.8％、2.8％、6.8％,批内变异系数为3.8％、12.7％、15.7％。样品中玉米赤霉烯酮掺合量与竞争间接酶联免疫吸附试验检出量有良好相关性(r≥0.9996)。     

用BRL-3A条件培养基培养鸡胚胎干细胞的初步研究

以CEF细胞、MEF细胞和STO细胞为饲养层,用:BRL-3A条件培养基培养寿光鸡X期胚盘细胞,并对获得的细胞克隆进行了鉴定,证明其具有ES细胞的克隆形态、PAS染色阳性、AKP染色阳性、能分化为多种类型的细胞、能参与受体胚胎的发育并形成羽色嵌合体.实验结果表明:BRL-3A条件培养基能促进鸡ES细胞克隆的形成,用饲养层比不用饲养层更有利于克隆的形成,且STO细胞和MEF细胞饲养层的培养效果要好于CEF细胞饲养层(P<0.05);挑克隆传代法比全消化法更有利于克隆的形成和未分化状态的维持(JP<0.05).     

小鼠白血病抑制因子基因的克隆、表达及其对维持鸡原始生殖细胞未分化状态的影响

为了探索鸡原始生殖细胞(Primordial germ cells,PGCs)适合的培养体系,我们在已构建的分泌型真核表达载体pSecTag-mlif(sp-)的基础上,通过脂质体介导将mlif转染到鸡PGCs和鸡胚胎成纤维(Chicken embryonic fibroblast, CEF)细胞中,48 h后收集细胞上清液,蛋白印迹均检测到小鼠白血病抑制因子(mLIF)的表达.以CEF细胞作饲养层,分七组来培养鸡PGCs,结果发现四组和五组培养的PGCs生长状态最好,三组传代后开始2-3天生长状况较好,3天后克隆周围有明显的分化现象.本实验将已构建了含mLIF基因的分泌型真核表达载体成功地瞬时转染到PGCs和CEF细胞中,且表达的mLIF具有维持鸡PGCs未分化状态的功能.     

人包皮传代细胞作为滋养细胞对人杂交瘤克隆生长的促进作用

用小鼠腹腔细胞等作为滋养细胞培养人杂交瘤,国内外均有报道。我们曾建立了人包皮传代细胞株。本文利用我们所建立的人包皮细胞做滋养细胞,并与小鼠腹腔细胞及不加滋养细胞的空白作对照,观察人包皮细胞对杂交瘤克隆生长的影响。 将人包皮细胞和小鼠腹腔细胞分别接种同一96孔板,次日将两株不同的人杂交瘤细胞1B4及1C8稀释至每毫升含10个或50个细胞,每孔接种0.1ml,使每孔分别含1个及5个细胞,经不同时间观察每孔杂交瘤克隆的生长情况,记录有杂交瘤生长的孔数。表1结果表明,     

Cerumen phenotypes in certain populations of Eurasia and Africa

Cerumen polymorphism was studied in several populations of Eurasia and Africa. The frequencies of dry cerumen were shown to be high in Mongoloid populations and low among Europoids. Intermediate frequencies were found among peoples of subequatorial Africa. Special attention is paid to the potential for using this marker in population and anthropological studies.     

人血清脂蛋白与糖胺聚糖及人主动脉蛋白聚糖的相互作用

本文报道了在[Ca~(2+)]=30mmol/L时,人血清或人血清脂蛋白与各种糖胺聚糖(GAG)及人主动脉两种蛋白聚糖(PG)的相互作用。GAG与血清的作用能力为6—硫酸软骨素(C6—S)>肝素(Hep)>4—硫酸软骨素(C4—S)>透明质酸(HA)>硫酸皮肤素(DS)。极低密度脂蛋白(VLDL)及低密度脂蛋白(LDL)可与肝素作用形成不溶性复合物,而高密度脂蛋白(HDL)则不能。人主动脉硫酸软骨素—PG(CS—PG)、硫酸皮肤素—硫酸软骨素—PG(DS—CS—PG)与血清形成不溶性复合物的曲线类型不同,后者的类型似有利于DS—CS—PG与血清脂蛋白结合从而使之在动脉壁沉积。     

基因治疗与人类健康

随着分子生物学及基因工程技术的迅猛发展 ,基因治疗已经成为治疗人类疾病的重要方法之一 ,同时也是维护人类健康最有发展前景的手段之一。诸如遗传病、肿瘤、和传染病与心血管病的基因治疗。遗传免疫方面 ,病毒性疾病和肿瘤的基因治疗 ,如将病毒抗原基因 (HBsAg)及一些肿瘤抗原基因 (CEA)直接注入人体内而产生抗体 ;人类亚健康状态 ,如肥胖、秃顶、疲劳、衰老等的基因治疗。然而基因治疗目前仍面临着许多困扰 ,如基因治疗的有效性、安全性、及社会伦理等诸多问题 ,因此在临床实际应用中要慎之又慎。只有对基因治疗合理规范和正确引导并遵循伦理原则 ,才能最终推动现代医学的发展。     

Generation of Induced Pluripotent Stem Cells with High Efficiency from Human Umbilical Cord Blood Mononuclear Cells

Human induced pluripotent stem cells （iPSCs） hold great promise for regenerative med- icine. Generating iPSCs from immunologically immature newborn umbilical cord blood mononu- clear cells （UCBMCs） is of great significance. Here we report generation of human iPSCs with great efficiency from UCBMCs using a dox-inducible lentiviral system carrying four Yamanaka factors. We generated these cells by optimizing the existing iPSC induction protocol. The UCBMC-derived iPSCs （UCB-iPSCs） have characteristics that are identical to pluripotent human embryonic stem cells （hESCs）. This study highlights the use of UCBMCs to generate highly functional human iPSCs that could accelerate the development of cell-based regenerative therapy for patients suffering from various diseases.     

Primers Designed for Amplification of Echinococcus multilocularis DNA Amplify the DNA of Encephalitozoon-Like Spores in the Polymerase Chain Reaction

ABSTRACT. Microsporidian spores were developed from cells which were grown in vitro from a human liver lesion which was due to larval Echinococcus multilocularis . The microsporidian spores developed in the same fashion as an Encephalitozoon cuniculi . The Encephalitozoon -like spores were completely separated on Percoll gradients. The separated spores contained DNA capable of amplification by two different primer sets designed for the polymerase chain reaction (PCR) of E. multilocularis DNA. However, the cell DNA from which microsporidium developed was thoroughly insensitive to the PCR using the E. multilocularis primer sets. The results strongly suggested that Encephalitozoon should be taken into consideration, when DNA isolated from larval E. multilocularis is analyzed.     

Assessment of the human sperm acrosome reaction using concanavalin A lectin

A method for assessment of the human sperm acrosome reaction is reported using fluorescein isothiocyanate (FITC)-conjugated Concanavalin A (ConA). The technique involved labelling prefixed spermatozoa, where only those spermatozoa that showed a complete loss of the acrosome bound FITC-ConA to the acrosomal region. Competitive sugar binding studies demonstrated that binding of ConA lectin to the acrosomal area of human spermatozoa was inhibited in the presence of 0.2 M D-mannose. Staining with the supravital stain Hoechst 33258 (H258) concomitantly with FITC-ConA allowed determination of only those spermatozoa that had undergone a true and not degenerative acrosomal loss. Incubation of human spermatozoa with 0, 1, 5, and 25 microM calcium ionophore, A23187, for 60 min demonstrated that changes in acrosomal status due to the different treatment protocols may be determined by the dual-staining method. Electron microscopy studies revealed that gold-conjugated ConA bound specifically to the surface of the inner acrosomal membrane of acrosome-reacted spermatozoa. A significant correlation (r = +.97) between transmission electron microscopy (TEM) and FITC-ConA labelling methods of acrosomal status assessment was achieved. The simple ConA labelling procedure reported here therefore provides a reliable method for quantitation of the physiological acrosome reaction of a population of human spermatozoa.     

Number of siblings and children of short and long living individuals

The work aimed at grasping eventual relations between the grandparents' life length and the number of their siblings and children. A poll of 2800 students of Wroclaw higher education institutions (1023 men and 1777 women) from 18 to 26 years of age was conducted. Information concerning the number of siblings and children of the examined students' grandparents, and in case of the dead—their age at the moment of death, were used. The material was divided into two categories (using death—rate tables and death age median: short living (SL) and long living (LL). The examined relations were analyzed separately for mothers' mothers (MM), fathers' mothers (FM), mothers' fathers (MF) and fathers' fathers (FF) of the students combined these groups were also analysed. Number of siblings in the families of grandparents of the same categories of longevity generally doesn't differ significantly for each group of grandparents. Short living grandparents have significantly fewer siblings than the long living ones. Number of children, similarly as the number of siblings, is also higher in case of long living individuals. The described relation shall probably be sought in the so—called protective role of the family.     

Reflections on the main events and changing factors in the evolution of human behaviour. Present problems of change in human beings from an evolutionary perspective

In this paper the authors analize the main physical and cultural changes of human species from its first moments to the future with an evolutionary perspective. They consider physical items like bipedalism, fire, language or tool production, but also some cultural ones like burials, art or changes on the prehistoric group’s economy, food production, etc.