EB病毒4种反义寡核苷酸片段对鼻咽癌SUNE—1细胞株的影响

用不同浓度的ＥＢ病毒４种（ＢＨＲＦ１、ＤＮＡ酶、核抗原－１及胸苷激酶）反义寡核苷酸片段,（ａｎｔｉｓｅｎｓｅ ｏｌｉｇｏｄｅｏｘｙｎｕｃｌｅｏｔｉｄｅ,ＡＳＯ）分别与含１０％胎牛血清或不含胎牛血清的人人低分化鼻咽癌细胞株（ＳＵＮＥ－１）共同培养４８ｈ。结果仅适当浓度的ＢＨＲＦ１－ＡＳＯ能引起无血清培养的ＳＵＮＥ－１细胞株增殖明显受抑制,透射电镜及琼脂糖凝胶电泳检测结果发现ＳＵＮＥ－１细胞发生调亡有     

磷酸化对人肝癌细胞株SMMC-7721转醛醇酶活性的影响

目的:研究人肝癌细胞株SMMC-7721中转醛醇酶(Transaldolase,TAL)活性变化与磷酸化的关系。方法:SMMC-7721细胞培养,细胞周期同步化后分组加不同浓度(10%、1%)胎牛血清孵育,分别测定不同孵育时间(1、2、4、8h)TAL酶活性;Western blot检测TAL蛋白表达水平。磷酸酶消化法检测TAL活性变化与磷酸化的关系。结果:①人肝癌细胞株SMMC-7721中TAL活性在10%胎牛血清培养时明显高于1%胎牛血清,2、4、8h显著性差异(P<0.05)。②不同浓度(10%.1%)胎牛血清孵育2、4、8hTAL蛋白表达水平无显著性差异。③磷酸化消化后TAL活性,与消化前相比呈显著性下降。结论:人肝癌细胞株SMMC-7721中增加血清浓度明显提高TAL活性,酶活性变化与TAL蛋白表达水平无关,提示活性调节发生在蛋白翻译后修饰,其中磷酸化起重要作用。     

EB病毒编码的潜伏膜蛋白1参与鼻咽癌恶性演进的转录调控实验研究

探讨EB病毒潜伏膜蛋白1(LMP1)激活激活蛋白1(AP1),和核转录因子(NF-κB)在鼻咽癌细胞SUNE-1及亚细胞株恶性演进中的作用.运用报告基因法和凝胶电泳迁移率法(EMSA)分析AP1和NF-κB反式激活活性和DNA结合活性,蛋白质印迹检测蛋白质表达;裸鼠致瘤实验结合组织制片研究瘤细胞的成瘤和转移能力. 结果显示恶性程度不同的SUNE-1亚细胞株的反式激活活性、DNA结合活性、LMP1蛋白表达及c-Jun氨基端激酶(JNK)活性均存在明显差异,且与细胞恶性程度正相关.这些结果提示LMP1活化AP1和NF-κB的信号通路参与了鼻咽癌细胞SUNE-1的恶性演进过程.     

旋毛虫肌幼虫细胞传代培养及超微结构观察

消化、分离观察旋毛虫(Trichinella spiralis)肌幼虫,获得肌幼虫细胞,用含10%胎牛血清的RPMI-1640培养液培养原代细胞,胰酶(含0.02?TA)消化法进行传代,透射电镜观察培养细胞超微结构,用多重PCR鉴定培养细胞。结果表明,在培养24～72h原代细胞开始贴壁,7～8d形成单层细胞,细胞间融合现象不明显,10～12d传一代。透射电镜显示旋毛虫细胞核为椭圆形,核膜、核仁清晰,核内染色质较丰富,胞浆含丰富的线粒体。细胞主要有两种类型:椭圆形和多角形,以椭圆形为主。多重PCR扩增培养细胞DNA,可见1条与旋毛虫肌幼虫DNA扩增产物相同的条带(173bp)。结果表明,旋毛虫肌幼虫细胞可在含10%胎牛血清的RPMI-1640培养液中传代培养。     

用于杂交瘤繁殖的血清补充物

胎牛血清(FBS)已成为用于动物细胞(包括杂交瘤)培养的精选的血清。较廉价的其它血清诸如成牛血清和马血清,它们所含的高浓度免疫球蛋白可能干扰某些筛选测定或单克隆抗体的纯化。为了提供胎牛血清的低廉替代物,研制了一种缺少免疫球蛋白 G 的成牛血清(MBS)。MBS 的免疫球蛋白量非常低(每毫升低于150微克),对血细胞凝集作用、酶标测定和免疫荧光筛选没有干扰。现已用它来进行杂交瘤的常规培养,并产生了与胎牛血清可比拟的结果。然而,结果表明:MBS 不适宜杂交瘤的克隆和融合,但有利于需要大量培养基的克隆繁     

大鼠椎间盘髓核细胞体外凋亡模型的建立

目的建立大鼠椎间盘髓核细胞体外凋亡模型。方法为了充分模拟退变椎间盘内营养缺乏的微环境,本实验分别采用含1%,3%,5%,8%,10%胎牛血清的DMEM培养基培养椎间盘髓核细胞,筛选最佳促凋亡浓度,分别检测髓核细胞凋亡率、凋亡相关蛋白Bax、bcl-2、caspase-3酶的表达、细胞增殖曲线及免疫荧光分析。结果流式细胞仪测得髓核细胞凋亡率随着胎牛血清(FBS)浓度降低而升高,3%FBS为最有效诱导凋亡浓度;Western blot示Bax、caspase-3酶表达在3%FBS组明显高于10%FBS组,同时bcl-2表达下降;CCK-8检测结果显示含3%FBS的培养基内,随着凋亡率的增长,髓核细胞增殖的速率越来越慢;免疫荧光分析3%FBS组FAS表达量明显比10%FBS组增高。结论 3%FBS能诱导髓核细胞发生凋亡,最终会导致细胞功能丧失,caspase家族参与并执行了这一过程。     

土壤可培养细菌DNA的提取及RAPD条件的优化*

以改进的CTAB 溶菌酶 蛋白酶K裂解法抽提土壤可培养细菌总DNA ,直接进行随机扩增多态DNA (RAPD)分析。分别测试了不同浓度镁离子、dNTP、模板DNA、引物、DNA聚合酶及牛血清白蛋白对反应结果的影响 ,通过各因子的组合研究 ,确定了土壤可培养细菌遗传多样性分析的稳定的RAPD反应体系。     

金霉素单克隆抗体的制备及检测方法的建立

采用羰基二咪唑法,将半抗原金霉素(AM)分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联制备金霉素免疫抗原AM-BSA和检测抗原AM-OVA,通过紫外光谱扫描检测偶联产物。采用细胞杂交瘤技术,制备抗金霉素单克隆抗体杂交瘤细胞株,建立了金霉素竞争ELISA检测方法,其灵敏度达到50ng/ml,且呈现良好的线性关系(r=0.9812),并且与其他抗生素无交叉反应。     

土壤可培养细菌DNA的提取及RAPD条件的优化

以改进的CTAB-溶菌酶-蛋白酶K裂解法抽提土壤可培养细菌总DNA,直接进行随机扩增多态DNA(RAPD)分析。分别测试了不同浓度镁离子、dNTP、模板DNA、引物、DNA聚合酶及牛血清白蛋白对反应结果的影响,通过各因子的组合研究,确定了土壤可培养细菌遗传多样性分析的稳定的RAPD反应体系。     

一种简单的人脐带间充质干细胞分离培养方法

目的 建立一种简单的人脐带间充质干细胞分离培养方法.方法 取新鲜脐带,剪成5 cm长的小段,直接剪碎为糊状,加入含10％胎牛血清的DMEM/F12在培养瓶中培养,光学显微镜下观察细胞的生长特征,运用流式细胞仪检测分析细胞的抗原标志表达,并检测其体外多向分化潜能.结果 运用不剥离血管组织、不用酶消化的组织贴块培养法可以从...     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism