中国鄂温克、鄂伦春、达斡尔族永生细胞系的建立与保存

本采用ＥＢＶ（Ｅｐｓｔｅｉｎ－Ｂａｒｒ Ｖｉｒｕｓ）上清液转化Ｂ淋巴细胞,并加入环胞霉素Ａ（Ｃｙｃｌｏｓｐｏｒｉｎｅ Ａ）抑制Ｔ淋巴细胞,成功地对中国东北地区鄂温克族、鄂伦春族及达斡尔族的部分个体建立了永生细胞系,其中鄂温克族４９株,鄂伦春族４０株,达斡尔族５１例,总计１４０株。永久保存我国特有民族的基因组,为分析其遗传学差异奠定了基础。     

建立海南黎族永生细胞库

采用ＥＢ病毒转化外周血Ｂ淋巴细胞加环胞霉素Ａ 法,建立了含５４ 株海南黎族永生细胞库（男性５６％ ,女性４４％ ）,有亲缘关系者占１３％ 。供血者身体健康,三代无与其他民族或支系通婚史。     

中国鄂温克、鄂伦春、达斡尔族永生细胞系的建立与保存

本文采用EBV（Epstein?Barr Virus）上清液转化B淋巴细胞,并加入环胞霉素A（Cyclosporine A）抑制T淋巴细胞,成功地对中国东北地区鄂温克族、鄂伦春族及达斡尔族的部分个体建立了永生细胞系,其中鄂温克族49株,鄂伦春族40株,达斡尔族51株,总计140株。永久保存我国特有民族的基因组,为分析其遗传学差异奠定了基础。 Abstract：The immortal lymphoblastoid cell lines were established by EBV transformation of B cells and addition of cyclosporin A to inhabit the activity of T cells.In the present study ,140 immortal cell lines of the Ewenki,the Oroqen and the Daur ethnic groups in the Northeast China were established .This is an important part of the research of human genome diversity for the exploration of the origin and progression of different ethnic groups ,and also provide enough research materials for further studies.     

利用染色体G显带技术鉴定人永生细胞实验条件初探

目的:探索利用染色体G显带技术鉴定人永生淋巴细胞的最佳实验条件。方法:在常规染色体制片技术的方法和手段的基础上,分别观察不同PHA浓度和培养时间对永生淋巴细胞分裂增殖的影响;然后,选择不同浓度秋水仙素处理细胞,观察染色体分散程度和中期分裂相的形态。结果:在永生淋巴细胞培养48h,PHA终浓度为0.1mg/mL,秋水仙素终浓度为0.04μg/mL,且其作用时间为1.5~2h的条件下,所获得的细胞染色体标本分裂相较多、染色体较长、分散较好、利于带型分析。结论:利用染色体G显带技术鉴定人永生淋巴细胞提供了技术保证,降低了实验成本,并且缩短了实验时间。     

线粒体遗传疾病细胞模型的构建:永生淋巴细胞系和转线粒体细胞系

线粒体基因组突变会引发多种线粒体遗传疾病。永生淋巴细胞系和转线粒体细胞系是线粒体遗传疾病研究的重要工具之一。永生淋巴细胞系使特殊遗传信息在细胞水平得以永久保存。转线粒体细胞系技术的发展为线粒体遗传疾病的分子机制研究提供了体外细胞平台。早期转线粒体细胞系的胞质供体直接来源于未进行永生化的患者各组织细胞或血小板。本文结合永生化手段,以线粒体4401AG为例,详细介绍了从冻存全血构建永生化淋巴细胞,然后再与ρ0 206一起构建转线粒体细胞系的原理、方法和技术路线,并将两种细胞模型的构建归纳为4个步骤:(1)永生淋巴细胞构建;(2)转化;(3)筛选;(4)鉴定。为真实反应线粒体突变的功能,本文对构建的永生淋巴系和转线粒体细胞系的线粒体突变位点、拷贝数以及转线粒体细胞系的细胞核型进行了分析与鉴定,选取了各参数一致的细胞系用于贮存与分析,以减少实验误差对后续突变位点功能研究的影响。两种细胞模型在细胞水平上为诠释线粒体遗传疾病的分子机制发挥了重要作用,但在具体应用中需注意它们的局限性,尤其是组织特异性的线粒体疾病。     

含HTNV S基因重组痘苗病毒在HFRS患者B淋巴母细胞系中的表达

采用Ficoll密度梯度离心法(淋巴细胞分离液)分离肾综合征出血热(HFRS)患者外周血单个核细胞(PBMC),并用EB病毒(EBV)感染B淋巴细胞,建立永生化的B淋巴母细胞系(B-LCL).然后,用含汉滩病毒(Hantaan virus ,HTNV) S基因的重组痘苗病毒感染B-LCL,应用间接免疫荧光检测核衣壳蛋白的表达.结果表明,B淋巴细胞经EBV感染4周左右,可形成永生化B-LCL.成功转化后的B-LCL,体积增大,且增殖的淋巴细胞积聚成团.汉滩病毒S基因在B-LCL中能有效表达核衣壳蛋白.含S基因的重组痘苗病毒感染的B-LCL可用作HTNV核衣壳蛋白特异性CTL活性研究的靶细胞.     

人卵巢浆液性囊腺癌永生化细胞系BUPH∶OVCA-3的建立及其生物学特性

介绍人卵巢浆液性囊腺癌永生化细胞系的建立 ,研究其生物学特性 .以卵巢浆液性乳头状囊腺癌的腹水细胞为材料 ,进行体外培养 .将永生化基因———SV4 0T抗原基因转染第 2代细胞 ,得到永生化细胞系 .通过光学显微镜、生长曲线测定、染色体分析、双层软琼脂培养、裸鼠接种、免疫组化等 ,研究其生物学特性 ,并与其来源细胞的生物学特性进行比较 .建立了一株人卵巢浆液性囊腺癌永生化细胞系 ,命名为BUPH∶OVCA 3,现已传至 6 0余代 .其生物学特性为 ,细胞生长旺盛 ;具有人体恶性细胞的核型特征 ;细胞恶性度较低 ,不具有集落形成能力及裸鼠接种致瘤性 ;除较未永生化细胞生长速率增快 ,饱和密度增加外 ,仍保留上皮细胞的分化表型 .结果表明 ,BUPH∶OVCA 3为一株恶性度较低的人卵巢浆液性囊腺癌永生化细胞系 ,保留其来源细胞的生物学特性 ,可作为研究恶性度较低的卵巢上皮癌的体外模型     

含HTNV S基因重组痘苗病毒在HFRS患者B淋巴母细胞系中的表达

采用Ficoll密度梯度离心法(淋巴细胞分离液)分离肾综合征出血热(HFRS)患者外周血单个核细胞（PBMC),并用EB病毒(EBV)感染B淋巴细胞,建立永生化的B淋巴母细胞系(B—LCL)。然后,用含汉滩病毒(Hantaan virus,HTNV)S基因的重组痘苗病毒感染B—LCL,应用问接免疫荧光检测核衣壳蛋白的表达。结果表明,B淋巴细胞经EBV感染4周左右,可形成永生化B—LCL。成功转化后的B—LCL,体积增大,且增殖的淋巴细胞积聚成团。汉滩病毒S基因在B—LCL中能有效表达核衣壳蛋白。含S基因的重组痘苗病毒感染的B—LCL可用作HTNV核衣壳蛋白特异性CTL活性研究的靶细胞。     

带有逆转录病毒的恶性T淋巴细胞株的建立

从一例患神经系统疾病病人的外周血淋巴细胞中建立了一株恶性Ｔ淋巴细胞株ＣＭ－１并研究了它的生物学特性。用过滤的ＣＭ－１细胞的培养上清,可命名多发性血管硬化症病人的淋巴细胞转化恶性Ｔ淋巴细胞,由此建立ＣＭ－２细胞株。用ＣＭ－１和ＣＭ－２细胞皮下接种裸鼠,都能使裸鼠产生弥漫性恶性淋巴瘤。电镜下见到了类似于Ｃ型逆转录病毒的颗粒,逆转酶活性检测阳性。血清学和基因检测表明ＣＭ－１和ＣＭ－２中不存在本室常用的其     

人乳头瘤病毒16型长控制区上YY1结合位点的破坏可增强病毒诱导细胞永生化能力

细胞转录调节因子 Ｙ Ｙ１ 可抑制人乳头瘤病毒１６ 型（ Ｈ Ｐ Ｖ １６） 癌基因启动子 Ｐ９７ 的活性, Ｙ Ｙ１ 位点的突变和缺失不仅可诱导 Ｐ９７ 活性增强而且可在全基因组内增强 Ｅ６ 癌基因转录,同时使病毒对啮齿类动物纤维细胞的转化能力增强。为了观测人乳头瘤病毒１６ 型长控制区（ Ｈ Ｐ Ｖ１６ Ｌ Ｃ Ｒ） 序列上 Ｙ Ｙ１ 蛋白特异性结合位点破坏在完整基因组范围内对人原代包皮角源细胞永生化能力的影响,将 Ｈ Ｐ Ｖ １６ Ｙ Ｙ１ 位点突变株和野毒株转染至人原代包皮角源细胞。筛选结果表明,突变株可诱导形成永生化细胞,永生化能力明显高于野毒株。对４ 株永生化细胞系 Ｄ Ｎ Ａ检测发现,均含有呈整合状态的 Ｈ Ｐ Ｖ １６ Ｄ Ｎ Ａ,其中３ 株的 Ｅ１／ Ｅ２ 区域有缺失。 Ｒ Ｎ Ａ 检测显示,４株细胞内均有 Ｅ６／ Ｅ７ ｍ Ｒ Ｎ Ａ 的转录。这表明, Ｈ Ｐ Ｖ １６ Ｌ Ｃ Ｒ 上 Ｙ Ｙ１ 蛋白特异性结合位点的破坏,可在完整基因组范围内增强病毒使人原代包皮角源细胞永生化的能力。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.