中国蝴蝶新种记述（Ⅱ）（鳞翅目）

记载采自河南省、四川省和云南省的中国蝴蝶8新种,模式标本保藏在河南省科学院和河南师范大学生物系。     

基因多态性（下）（Polymorphisms PartⅡ）

以绘制人类基因单体型图为目标的国际Hapmap计划也接受尾声。 基于SNP信息的基因组药理学已经开始实际应用。 个性化医疗在现实医疗中正在生根发芽。 接着上回由东大医科研的中村祐辅教授继续进行讲解。[编者按]     

中国蝴蝶新种记述（Ⅰ）（鳞翅目）

记载采自湖南省的中国蝴蝶3新种,模式标本保存在湖南省东安县第一中学。     

正（误）、宜用（不宜用）的部分生理学名词（术语）

在杂志和书刊上经常见到一些误用或不宜用的生理学术语,自然科学专业名词(包括医学名词)一律以全国自然科学名词审定委员会公布的名词为标准。考虑到有些专业名词多年来未审定,根据近年来科学出版社、高等教育出版社的图书编写要求,中华医学杂志和中国药理学报的投稿要求,     

花之美（下）

“等闲识得东风面,万紫千红总是春”。无论是冰清玉洁的玉兰、奔放的玫瑰、还是悠然的紫罗兰,抑或清风傲骨的梅花,都是文人作品中频频出现的“座上客”。为什么对花朵的赞美早已成为一种追求真善美的文化作品而存在呢?毫无疑问,花朵所展现的那种健康之美以及那种超群的生命力不得不让人们为之赞叹。花谢之时,其风华就不在了吗?其实,生命的轮回往复正是一种自然规律的表现,逝去是一种必然。生如夏花之绚烂,死如秋叶之静美,当花朵不再娇艳欲滴,那正是果实成熟之时,恰似一个孕育着新生命的母亲,将自己的浮华美丽渐渐褪去,用自己的青春换来一个新生命的到来。这即是植物生命力的最直接表现,这就是花之美。     

贺信（英文）

<正>~~     

表观遗传学解析（上）（基因组概况之一）

由哺乳类所代表的多细胞生物，由于来自一个受精卵（也就是说只有一个细胞），对于体内的任何组织的任何细胞来说，除免疫细胞等特殊情况之外，其中的任何细胞的基因组序列都相同。[第一段]     

人类分子遗传学（原书第三版）（Human Molecular Genetics3）（含光盘）

人类基因组计划于2003年发表了人类基因组序列的“完成”版，这是人类科学史上的一个里程碑。根据人类基因组许多新的发现，英国Tom Strachan和Andrew P．Read两位教授对其编著的《人类分子遗传学》第三版进行了全新的修订。     

Immunochemical detection of cell cycle synchronization in a human erythroleukemia cell line, K562

Cells of the human erythroleukemic line K562 can be induced by manipulation of culture conditions to arrest within the G1 phase of the cell cycle, and subsequently to enter S phase synchronously after release from G1. Cell cultures subjected to serum deprivation and hydroxyurea (HU) treatment demonstrated less than 5% of the cells to be in S phase. Four hours after release from HU, 63% of the cells were in S phase, as detected by immunofluorescent staining. This protocol offers a method for synchronization of K562 cells at the G1/S border and a technique for detection of S-phase cells without the use of radioisotopes or flow cytometry instrumentation.     

IL——3和羟基脲协同诱导K562细胞凋亡

The effect of IL-3 and hydroxyurea on human erythroleukemia cell line (K562 cells) was demonstrated by using the electro-microscopy and flow cytometry. Our data showed that neither IL-3 nor hydroxyurea could induce the apoptosis of K562 cells alone. However, the IL-3 and hydroxyurea could induce the apoptosis of K562 cells cooperatively. Analysis with flow cytometry showed that the percentage of apoptotic cells was about 31.90% after K562 cells were induced by IL-3 and hydroxyurea cooperatively for 5 days, and the sub-G1 peak (apoptotic peak) was detected in the induced K562 cells. Meanwhile, the percentage of S-phase in the IL-3 and hydroxyurea induced K562 cells was increased, and the proliferation of the induced K562 cells was inhibited significantly. Furthermore, the IL-3 and hydroxyurea induced K562 cells showed chromatin condensation with regular crescents at the nuclear edges and apoptotic bodies. It suggested that IL-3 could enhance the sensitivity of K562 cells to hydroxyurea and the apoptosis of K562 cells could be induced by IL-3 and hydroxyurea cooperatively.     

IL-3和羟基脲协同诱导K562细胞凋亡

本文应用流式细胞分选仪和电子显微镜研究了IL-3和羟基脲对人红白血病细胞株(K562细胞)凋亡的影响.结果显示IL-3和羟基脲分别诱导K562细胞,不能引起细胞凋亡;而IL-3和羟基脲协同诱导K562细胞,可以引起细胞凋亡.用流式细胞仪检测到IL-3和羟基脲协同诱导K562细胞后,DNA含量低于二倍体的细胞数达31.90%,并产生明显的凋亡小峰.同时,IL-3和羟基脲协同诱导K562细胞,可抑制细胞周期中的S期,阻止细胞从S期进入G2/M期,使细胞周期延长,对K562细胞的生长和增殖具有抑制作用.在电镜下可观察到IL-3和羟基脲协同诱导的K562细胞,出现典型的凋亡细胞形态,细胞核内染色质浓缩、凝聚,紧靠在核膜边沿,形成新月形或环状的染色质结构,产生凋亡小体.提示IL-3和羟基脲具有协同效应,IL-3可提高K562细胞对羟基脲的敏感性,并可协同羟基脲诱导K562细胞凋亡.     

Effects of hydroxyurea on monoclonal antibody production induced by anti-mIgG and LPS stimulation on murine B cell hybridomas

Chemical treatment with hydroxyurea (HU) has been selected as a simple and low cost strategy to generate a cell population enriched for the G1 phase. After the chemical treatment with HU, cells were stimulated with anti-mIgG to test if the positive effects of anti-mIgG on CD40 expression and specific IgG2a production rate were improved upon a cell population with a higher percentage of cells in G1 phase at the beginning of the cell culture. In addition, other treatments assayed in this work were the cell stimulation with Lipopolysaccharide (LPS) both before and after the HU treatment. It has been observed that the use of HU under conditions able to maintain the cells in viable state (0.1 mM for 20 h), has a negative effect on CD40 expression and specific IgG2a production rate induced by anti-mIgG. The positive effect of LPS on cell stimulation induced by anti-mIgG is reduced on cells treated with HU.     

The role of a 70 kDa nuclear protein in fetal haemoglobin synthesis in K562 cells

Abstract. Previously, we reported that a 70 kDa nuclear protein may regulate fetal haemoglobin gene expression in haemin treated K562 cells. To obtain further evidence of the specific role of this 70 kDa nuclear protein, we compared the nuclear fractions isolated from phenylacetate, hydroxyurea and haemin treated K562 cells. Both phenylacetate and hydroxyurea have been used to induce fetal haemoglobin synthesis in K562 cells. Cell growth was measured by biochemical events including DNA, RNA and protein synthesis. Differentiation of K562 cells was determined by both [³H]-leucine incorporation into fetal haemoglobin and scoring benzidine-stained positive cells. Unlike the haemin treated cells, phenylacetate and hydroxyurea induced growth arrest and increased fetal haemoglobin gene expression in K562 cells. After four days of treatment with phenylacetate and hydroxyurea more than 50% of the cells stained positive with benzidine. The SDS-Polyacrylamide gel electrophoretic analysis of nuclear proteins isolated from phenylacetate and hydroxyurea treated K562 cells showed that the 70 kDa protein was reduced in nuclear protein extract in both groups similar to haemin treated cells. These results suggest that the loss of the 70 kDa protein from a nuclear protein extract is not restricted to only haemin treated cells but also occurs in hydroxyurea and phenylacetate treated cells. Our results provide further evidence that the 70 kDa nuclear protein may be involved in regulating fetal haemoglobin expression through a negative control mechanism.     

Induction of apoptosis in K562 cells by jolkinolide B

Jolkinolide B, a bioactive diterpene isolated from the roots of Euphorbia fischeriana Steud, has various biological and pharmacological properties. In this study, the cytotoxicity of highly purified jolkinolide B was tested in human chronic myeloid leukemia (K562) and 2 other cell lines (human esophageal carcinoma Eca-109 and human hepatoma HepG2). The results indicate a significant decrease in the proliferation of all the 3 cell lines when treated with jolkinolide B for 24 h; the IC50 value of cytotoxicity was 12.1 microg/mL (for K562 cells), >50.0 microg/mL (for HepG2 cells), and 23.7 microg/mL (for Eca-109 cells). Further study of K562 cells involving fluorescence and transmission electron microscopy revealed characteristic apoptotic features, such as cell shrinkage, membrane blebbing, loss of microvilli, and nuclear condensation. Agarose electrophoresis of genomic DNA showed a typical fragmentation pattern for apoptotic cells. A kinetic cell-cycle analysis demonstrated that the cell cycle was arrested in the G1 phase. All these results suggest that the anti-proliferation effect of jolkinolide B on K562 cells is achieved by arresting the cell cycle in the G1 phase and subsequently inducing apoptosis.     

Expression of Sara2 human gene in erythroid progenitors

A human homologue of Sar1, named Sara2, was shown to be preferentially expressed during erythropoiesis in a culture stimulated by EPO. Previous studies, in yeast, have shown that secretion-associated and Ras-related protein (Sar1p) plays an essential role in protein transport from the endoplasmic reticulum to the Golgi apparatus. Here, we report the molecular analysis of Sara2 in erythroid cell culture. A 1250 bp long cDNA, encoding a 198 amino-acid protein very similar to Sar1 proteins from other organisms, was obtained. Furthermore, we also report a functional study of Sara2 with Real-time quantitative PCR analysis, demonstrating that expression of Sara2 mRNA increases during the initial stages of erythroid differentiation with EPO and that a two-fold increase in expression occurs following the addition of hydroxyurea (HU). In K562 cells, Sara2 mRNA was observed to have a constant expression and the addition of HU also up-regulated the expression in these cells. Our results suggest that Sara2 is an important gene in processes involving proliferation and differentiation and could be valuable for understanding the vesicular transport system during erythropoiesis.     

Antizyme1基因转染对K562细胞增殖与凋亡的影响

研究鸟氨酸脱羧酶抗酶蛋白对人红白血病K562细胞增殖、三氧化二砷（ As2O3）诱导凋亡时的影响。方法: 定点突变技术构建缺失frameshift位点的pEGFP-N1-AZ1-mutation重组表达载体。脂质体法转染K562细胞,通过G418筛选获得稳定表达antizyme1的K562pAZ1m细胞系。采用不同浓度的As2O3处理细胞,通过MTT法检测细胞增殖,流式细胞术分析细胞周期及凋亡变化。并通过RT-PCR方法检测antiyme1转染对cyclin D1和survivin基因表达的影响。结果：获得稳定表达antizyme1的K562-AZ1m细胞株后,其增殖能力明显减慢。CyclinD1基因表达降低,细胞主要停滞于G0/G1期。在 As2O3的诱导作用下,细胞凋亡增多,survivin基因表达降低。结论：AZ1基因能够抑制K562细胞增殖,通过对cyclinD1的负调控使细胞周期停滞于G0/G1期。并可能通过下调survivin表达来加强 As2O3对其的诱导凋亡作用     

Synchronization of human leukemic cells: Relevance for high-resolution chromosome banding

Summary The cell-cycle kinetics of synchronized K562 human leukemic cells and bone marrow cells from adults with acute leukemia were studied in order to develop more reliable methods for producing increased numbers of mitoses, particularly those with elongated chromosomes suitable for high-resolution banding. Parameters examined included DNA content, mitotic index (MI), and chromosome preparations. K562 cells synchronized with methotrexate (MTX), thymidine (Tdr), or hydroxyurea (HU) showed two-fold increases in peak MI. Optimal harvesting times after release from block were approximately 10.5, 12.5, and 14.5 h for MTX, HU, and Tdr, respectively. MTX was selected for studies with cells from patients. Cells from 7 of the 10 patients studied showed 4.4-fold increases in peak MI. The optimal harvesting time was 9.5 to 11.5 h after release from block, considerably later than the 6 h time previously assumed in studies using stimulated lymphocytes. Cells from the three remaining patients showed no increase in MI after synchronization: and the lack of response may have been related to the high proportion of cells in G₀+G₁ prior to MTX exposure. For both the K562 cell line and most patient specimens, the combination of synchronization with appropriate release times and short Colcemid exposure (10 min) resulted in substantially improved chromosome preparations.