用Epstein-Barr病毒转化技术建立分泌人单克隆抗体B淋巴细胞系的条件

用以去除T淋巴细胞的E—玫瑰花形成法中所用的羊红细胞(SRBC),其保鲜程度对EB病毒转化B淋巴细胞的结果有一定影响,用保存1天的SRBC者,转化细胞的抗体分泌时间较长(8～10周),并且能形成传代细胞系;用保存17天的SRBC,转化细胞的抗体分泌时间较短,未能无限传代。分离外周血单个核细胞时,保留自体粘附细胞作为饲养细胞,转化的细胞二个月的成活率可达75％,而不保留粘附细胞者,转化细胞成活率仅40％。能产生抗-HBc抗体的EBV转化细胞,自然分泌抗体时间一般不超过10周,高峰期在3周以内,第2周时最高。EB病毒感染后的转化细胞5天开始克隆,一般细胞难成活,在14天克隆的,成活率可达100％。21天开始克隆的转化细胞,免疫球蛋白自然分泌维持12周以上,而在35天开始克隆的细胞,维持分泌的时间都不超过10周。以未经γ线照射的小鼠腹腔渗出细胞作饲养细胞,可支持转化3周的细胞克隆扩增,克隆成活率可达100％,而对照组克隆成活率仅为64.7％。     

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。     

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.     

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。     

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.     

A highly sensitive fluorometric assay for determination of human coagulation factor XIII in plasma

Based on the iso-peptidase activity of human plasma FXIII, a novel fluorometric assay that determines FXIII concentrations in human plasma below 0.05 IU/ml is introduced. We considered a peptide sequence derived from alpha(2)-antiplasmin (n =12) to yield high sensitivity. Peptide Abz-NE(Cad-Dnp)EQVSPLTLLK exhibits a K(m) value of 19.8+/-2.8 microM and is used in a concentration of 50 microM. The assay design is suitable for measurements in cuvettes (1 ml volume) as well as for the microtiter plate (MTP) format (0.2 ml volume). It provides linear dose-response curves over a wide range of FXIII concentrations (0.05-8.8 IU/ml). The assay was validated with respect to precision, detection and quantitation limits, accuracy/specificity, linearity, and range. A comparison of the fluorometric assay with the photometric assay for FXIII determinations in plasma pools as well as single donor plasma revealed suitability of the fluorometric assay for FXIII determination in plasma of healthy individuals. FXIII concentrations in plasma samples of patients with severe FXIII deficiency are discussed in the context of FXIII antigen levels. These assays correlate well in the critical range below 0.1 IU/ml, whereas the photometric assay may overestimate residual FXIII activity in severe FXIII-deficient patients.     

Where the wild things are: environmental preservation and human nature

Environmental philosophers spend considerable time drawing the divide between humans and the rest of nature. Some argue that humans and our actions are unnatural. Others allow that humans are natural, but maintain that humans are nevertheless distinct. The motivation for distinguishing humans from the rest of nature is the desire to determine what aspects of the environment should be preserved. The standard view is that we should preserve those aspects of the environment outside of humans and our influence. This paper examines the standard view by asking two questions. First, are the suggested grounds for distinguishing humans from the rest of the environment viable? Second, is such a distinction even needed for determining what to preserve? The paper concludes that debates over whether humans are natural and whether humans are unique are unhelpful when deciding what to preserve.     

The prolonged half-lives of new erythropoietin derivatives via peptide addition

Erythropoietin, or Epo, is a hematopoietic cytokine that promotes erythropoiesis, and recombinant human Epo has been used in the treatment of anemia in various chronic diseases. Here, we have constructed novel Epo derivatives with prolonged half-lives by adding peptides to the carboxy terminus of Epo without using linkers. The fused peptides were selected from the carboxy terminal region of human chorionic gonadotropin (hCG) or human thrombopoietin (hTpo), which promote the proper folding, secretion, and stabilization of bioactive glycoproteins. Addition of these peptides did not interfere with secretion or receptor binding, and significantly increased the in vivo half-life of human Epo, as measured by intravenous administration in rats. The plasma half-life of the Epo constructs was longest when the carboxy terminal 28 aa of the beta subunit of hCG was added (Epo-CGC), a half-life that was slightly longer than NESP (Aranesp), which is the most effective Epo product in current clinical use. The transformation of four Ser glycosylation sites to Ala on the CGC sequence also lengthened the plasma half-life of Epo, indicating that the in vivo stabilizing effect of the hCG peptide was due to both structures within the peptide itself and its O-glycosylations. The application of the carboxy terminal half of hTpo also resulted in remarkably reduced elimination of the Epo chimera (Epo-TpC), possibly due to protection by the TpC sequence. The in vivo hematopoietic activity of Epo derivatives in mice was consistent with their pharmacokinetic profiles. Therefore, these derivatives with prolonged half-lives may provide opportunities for developing new Epo therapeutics with less frequent administration.     

用HLA基因频率计算人群间的遗传距离

本文用白细胞抗原(HLA)的基因频率按Nei氏法计算了中国11个民族和其他人种与国家的六个人群间的遗传距离,并按Sneath和Sokal氏聚类原则绘制了这些群体的系统树。得出的结果与用红细胞血型资料所计算的结果基本吻合,群体间遗传距离和群体分布的地理距离呈一定平行关系。     

The 2011 Human Liver Proteome Project (HLPP) Workshop September 5, 2011, Geneva, Switzerland

During the 10th HUPO Annual World Congress held in Geneva (Switzerland) from 4th to 7th September, a workshop on Human Liver Proteome Project (HLPP) Initiative took place. Four research groups presented their latest results from different ongoing projects. Later on, during the HLPP executive members' meeting, the status of current projects and the next possible steps to be taken were discussed.     

Presence of dUTPase in the Various Human Endogenous Retrovirus K (HERV-K) Families

Various retroviruses have been shown to encode dUTPase. The overall phylogeny of dUTPase is unclear, though. The human genome contains a significant amount of human endogenous retroviruses (HERV) representing fossilized sequences of ancient exogenous retroviruses. A few HERV families have been reported to harbor dUTPase domains. We surveyed the various HERV families for the presence of dUTPase and found that ancestors of all HERV-K families but one encoded dUTPase. With two exceptions phylogenetic analysis shows a monophyletic origin of dUTPase for the different HERV-K dUTPases. Sequences of consensus dUTPase domains suggest that the various exogenous ancestors of HERV-K once encoded active enzymes. Our analysis provides informations on dUTPase phylogeny and further shows that endogenous retroviruses provide important informations regarding retrovirus evolution.     

Report of the 9th HLPP Workshop October 2007, Seoul, Korea

The Human Liver Proteome Project is one of the Human Proteome Initiatives launched by Human Proteome Organization (HUPO). Major achievements of the project have been obtained under the efforts of international collaboration with all the participants since it was formally proposed in 2002. Its updated progresses were presented in the latest workshop held in conjunction with the sixth HUPO World Congress in October, 2007, Seoul, Korea. Furthermore, four topics related to the project as well as other initiatives were lively discussed among all the attendees.     

Human Phenol Sulfotransferase: Correlation of Brain and Platelet Activities

Phenol sulfotransferase (PST; EC 2.8.2.1) catalyzes the sulfate conjugation of phenolic and catechol neurotransmitters and drugs. The human blood platelet has been the most thoroughly studied source of PST because of the possibility that the regulation of the enzyme in this easily accessible tissue might reflect the regulation of PST in the CNS. The human brain and platelet contain at least two forms of PST, forms designated as thermostable (TS) and thermolabile (TL) PST. TS PST catalyzes the sulfate conjugation of micromolar concentrations of phenol and p-nitrophenol and TL PST catalyzes the sulfate conjugation of dopamine and other monoamines. This study was performed to determine whether individual variations in the activities of human platelet TS and TL PST reflect individual variations in cerebral cortical PST activities. PST activities were measured in platelets and in cerebral cortical tissue obtained from 15 patients with epilepsy during clinically indicated neurosurgery. There was a highly significant correlation between the activities of the TS form of PST in cerebral cortex and platelets of these patients (r = 0.940, p less than 0.001), but there was not a significant correlation between activities of the TL form of PST in the two tissues (r = 0.396, p greater than 0.14). In addition to variations in the level of enzyme activity, there are also wide individual variations in the thermal stability of platelet TS PST.(ABSTRACT TRUNCATED AT 250 WORDS)     

中国法医人类学三十年

法医人类学在上世纪80年代开始引起中国法医学界的重视。改革开放后, 经济发展人口流动增加, 全国范围内无名尸体案件增加, 对骨骼个体识别的需求日益迫切。为了满足案件侦察的需要, 国内学者开始引进国外的有关研究结果, 并开始使用国人的材料对中国人骨骼个体识别的方法进行了广泛深入的研究。本文对中国学者近30年来在中国人骨骼年龄、性别及身高等方面的研究进行了回顾, 对相关研究与国外情况进行了比较,对有关问题的未来研究方向进行了讨论。