多因素对昆明小鼠胚胎干细胞原代集落形成和后期增殖的影响

目的：研究细胞因子LIF、EGF、bFGF和肝素钠,以及氧分压、温度等多因素对昆明系小鼠胚胎干细胞（KM-ESC）原代集落形成和后期增殖的影响。方法：本研究选择LIF、EGF、bFGF、肝素钠、5%O2,20%O2和37℃、39℃作为研究条件。并检测不同情况下小鼠胚胎干细胞原代集落形成和后期增殖情况。结果：LIF对KM-ESC原代集落形成和后期增殖具有显著促进作用,极显著高于EGF、bFGF和肝素钠组（P〈0.01）。温度对KM-ESC原代集落形成和增殖具有显著影响,39℃条件下,原代集落形成率、直径和后期增殖显著高于37℃（P〈0.05）;而氧分压对KM-ESC原代集落形成无显著作用（P〉0.05）,但是对原代集落直径和后期增殖有一定促进作用,20%O2组显著高于5%O2组（P〈0.05）。结论：LIF、EGF、bFGF、肝素钠、39℃、20%O2对小鼠胚胎干细胞原代集落形成和后期增殖具有显著促进作用。     

小鼠孤雌胚胎干细胞集落的建立

ＥＳＴＡＢＬＩＳＨＭＥＮＴＯＦＳＴＥＭＣＥＬＬＣＯＬＯＮＩＥＳＦＲＯＭＰＡＲＴＨＥＮＯＧＥＮＥＴＩＣＡＬＬＹＤＥＲＩＶＥＤＢＬＡＳＴＯＣＹＳＴＳＯＦＭＯＵＳＥ小鼠孤雌胚胎干细胞集落的建立ＫｅｙｗｏｒｄｓＭｏｕｓｅ,Ｐａｒｔｈｅｎｏｇｅｎｅｔｉｃｅｍ...     

昆明小鼠胚胎干细胞滋养层制备条件的实验研究

目的:建立小鼠胚胎成纤维细胞(MEFS)滋养层,用于昆明小鼠胚胎干细胞的培养。方法:取妊娠13.5的胎鼠,采用组织消化法分离培养出原代成纤维细胞,对MEFs的生长形态、生长曲线及分裂指数进行观察;MTT法筛选丝裂霉素C(MMC)作用的最佳浓度和时间;取妊娠3.5d的囊胚在经MMC处理的饲养层上培养,观察胚胎干细胞集落生成情况。结果:MEFS为一种贴壁生长且增殖速度较快的细胞,第三代细胞增殖旺盛,第5代以后细胞开始变形并趋于衰老;MMC能抑制胚胎成纤维细胞的增殖,最佳的作用浓度和时间是10ug/ml作用2.5～4h,20ug/ml作用1-2.5h。妊娠3.5d小鼠囊胚在饲养层上培养能形成典型的"鸟巢"状干细胞集落,并可维持胚胎干细胞的正常形态且不发生分化。结论:这种方法制备的滋养细胞层适用于胚胎干细胞的培养。     

饲养层及细胞因子对胚胎干细胞的影响

不同动物胚胎干细胞的生长条件不同,因此筛选合适的饲养层细胞建立适宜的培养体系是胚胎干细胞培养中的关键环节。本文就不同的饲养层细胞及各种外源细胞因子的作用做了简要论述。     

昆明小鼠胚胎干细胞的体外分离培养

利用昆明小鼠13.5dpc的胚胎制备小鼠胚胎成纤维细胞,并以小鼠胚胎成纤维细胞作为饲养层;收集3.5dICR小鼠的囊胚和桑椹胚进行体外培养,筛选纯化ES细胞集落,使其稳定传代后,对其形态学和生物学性状进行初步鉴定,获得阳性细胞集落。     

小鼠胚胎干细胞的培养

目的:建立小鼠胚胎干细胞(embryonic stem cells,ES)的培养方法。方法:制备G418抗性的原代小鼠胚胎成纤维细胞,经丝裂霉素C处理后成滋养层细胞,将小鼠胚胎干细胞复苏后,应用含白血病抑制因子的ES细胞培养液,培养小鼠ES细胞,观察集落的生长情况,并在光镜下观察细胞形态。结果:小鼠胚胎成纤维细胞生长良好,ES细胞呈克隆状生长,且保持未分化状态。结论:建立了小鼠胚胎干细胞培养的有效方法,为下一步基因打靶奠定基础。     

小鼠胚胎干细胞与四倍体胚胎的嵌合

The oviducts of superovulated Kunming white females were flushed 44-46 hours after treatment with human chorionic gonadotropin to collect 1074 late two-cell-stage embryos.The embryos were placed twenty at a time between two platinum electrodes laid 1 mm apart in 0.3M mannitol in the electrode chamber.The blastomeres were fused by a short electric pulse(80V for 50μsec) applied by a pulse generator.Fusion of blastomeres was usually completed in 20-60minutes.After 25 hours of culture,most of the tetraploid embryos developed to the four-cell stage.Zonae pellucidae of 387 four-cell-stage tetraploid embryos were removed by treatment with acid Tyrode‘s buffer.The embryos were plated on an ES cell layer,After 40 hours of coculture,248 embryos aggregated with ES cells were collected and transferred into the uteri of twenty four 2.5-day pseudopregnant recipinets.Ten recipients were pregnant.but no live fetuses were born.Three pregnant recipients were routinely subject to a Caesarean section on day 18 of pregnancy and seven abnormal fetuses were obtained.The results demonstrate that ES cells derived from C57BL/6 mice are pluripotential to a certain extent.     

品系对小鼠胚胎干细胞分离效率的影响

为了充分利用小鼠胚胎干(ES)细胞,就必须从众多小鼠品系中分离ES细胞系。本研究通过传统的成纤维细胞饲养层法,从CD-1、129/Sv、C57BL/6J和129/Sv×C57BL/6J四种不同遗传背景的小鼠中分离得到12个ES细胞系,而从KM小鼠没有得到ES细胞系。所有的ES细胞系都具有典型的ES细胞特征,AKP染色呈阳性。从四种不同遗传背景的ES细胞系得到了包含多种组织的畸胎瘤;与桑椹胚聚合后,都得到了生殖系嵌合体。结果表明:品系对小鼠ES细胞的分离有显著影响,利用129小鼠以及包含129小鼠遗传背景的杂交小鼠都较容易分离ES细胞,由ES细胞得到生殖系嵌合体的效率在不同品系间有显著差异,从杂交ES细胞比近交ES细胞中更容易得到生殖系嵌合体。     

昆明白小鼠胚胎干细胞分离与体外培养

为探索昆明白小鼠胚胎干细胞建系方法,将受孕4.5天的昆明白小鼠囊胚用免疫手术法去除滋胚层,然后将内细胞团(ICM)接种于胎鼠成纤维细胞饲养层上培养,形成的胚胎干细胞样集落用胰蛋白酶-EDTA消化法传代,培养后进行相差显微镜观察及碱性磷酸酶染色。结果饲养层上生长的ICM细胞呈典型的ES样细胞集落,传至第8代碱性磷酸酶染色呈强阳性。实验表明免疫手术法适用于昆明白小鼠ES细胞建系,获得的细胞集落具有ES细胞的主要生物学性状。     

小鼠胚胎干细胞的培养

ＥＳ细胞的培养应满足促进细胞增殖和抑制细胞分化而保持其高度未分化潜能。我们用胚胎成纤维细胞作为饲养层,培养基ＤＭＥＭ中加白血病抑制因子的方法来培养ＥＳ细胞。培养的ＥＳ细胞呈集落状生长,细胞排列紧密,呈未分化状态。胚胎成纤维细胞作为饲养层是分离培养ＥＳ细胞最普遍使用而且有效的方法。本文还围绕ＥＳ细胞培养中的促进增殖和抑制分化这两个方面的影响因素进行了讨论。     

Effects of Murine Erythroleukemia Inducers on Mouse Erythroid Colony Formation in Culture

The effect of various agents which are known to increase the differentiation of Friend erythroleukemia cells was investigated in cultures of mouse bone marrow cells. N, N-dimethylacetamide (5 and 15 mM) and acetamide (60 mM) significantly increased the number of erythroid colonies observed. Tetramethylurea, dimethylformamide, pyridine N-oxide, and butyric acid were ineffective. Dimethylsulfoxide at a concentration of 1% significantly increased colony number in cultures of marrow cells obtained from male mice, but had no effect in cultures of female bone marrow cells.     

几种主要环境因子对布朗葡萄藻(Botryococcus braunii)光合作用的影响

利用测定净光合放氧速率的方法研究了光照强度、温度、 pH值、盐度对布朗葡萄藻Botryococcus braunii UTEX 572和B.braunii UTEX 2441两个品系的光合作用的影响.B.braunii UTEX 572的适宜光照强度范围400～1600 μmol·m-2·s-1,光饱和点在800 μmol·m-2·s-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH7.0;适宜盐度范围0～0.2 mol/L,最适盐度0.1mol/L.B.braunii UTEX 2441的适宜光照强度范围400～1600 μmol·m-2·s-1,光饱和点在400 μmol·m-2·s-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH 7.0;对盐度的适应范围较小,盐度升高,光合放氧速率明显下降.两个布朗葡萄藻净光合放氧速率随光照强度、温度、pH值和盐度变化的规律,表明布朗葡萄藻的基本生理生态学特征:适应于较强的光照强度、较高的温度、中性偏酸的环境和较低的盐度.对布朗葡萄藻基本生理生态学特征的了解,为培养条件的优化提供了依据.2个布朗葡萄藻品系对光强、温度、pH值和盐度变化的反应有所不同:与B.braunii UTEX 2441相比,B.braunii UTEX 572具有更高的光饱和点,适应更高的温度,对pH值变化有更宽的适应范围,适当提高盐度对其光合作用有促进作用,表明B.braunii UTEX 572在快速生长繁殖方面具有更大的潜力,这一研究结果为筛选适合于大量培养的优良藻种提供了依据.     

建立小鼠胚胎多能干细胞系（ES细胞系）的几个主要影响因素

本文系统地比较了影响建立小鼠胚胎多能干细胞系的几个主要因素。对825个胚的实验进行了分析,其结果表明:延迟囊胚的主要作用是有助于内细胞团(ICM)的增殖;高糖的DMEM有助于囊胚贴壁及ICM的增殖;不含丙酮酸钠的DMEM对ES细胞的集落形成和生长是有利的;饲养层对ES细胞的建系和培养是十分重要的,以新鲜制备的原代小鼠胚胎成纤维细胞作为饲养层对ES细胞的生长有明显的促进作用。     

几种主要环境因子对布朗葡萄藻(Botryococcus braunii)光合作用的影响

利用测定净光合放氧速率的方法研究了光照强度、温度、pH值、盐度对布朗葡萄藻Botryococcus braunii UTEX 572和B.braunii UTEX 2441两个品系的光合作用的影响。B.braunii UTEX 572的适宜光照强度范围400～1600μmol·m^-2·s^-1,光饱和点在800μmol·m^-2·s^-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH7.0;适宜盐度范围0～0.2mol/L,最适盐度0.1mol/L。B.braunii UTEX 2441的适宜光照强度范围400～1600μmol·m^-2·s^-1,光饱和点在400μmol·m^-2·s^-1附近;适宜温度范围25～35℃,最适温度30℃;适宜pH范围5.0～8.0,最适pH7.0;对盐度的适应范围较小,盐度升高,光合放氧速率明显下降。两个布朗葡萄藻净光合放氧速率随光照强度、温度、pH值和盐度变化的规律,表明布朗葡萄藻的基本生理生态学特征:适应于较强的光照强度、较高的温度、中性偏酸的环境和较低的盐度。对布朗葡萄藻基本生理生态学特征的了解,为培养条件的优化提供了依据。2个布朗葡萄藻品系对光强、温度、pH值和盐度变化的反应有所不同:与B.braunii UTEX 2441相比,B.braunii UTEX 572具有更高的光饱和点,适应更高的温度,对pH值变化有更宽的适应范围,适当提高盐度对其光合作用有促进作用,表明B.braunii UTEX 572在快速生长繁殖方面具有更大的潜力,这一研究结果为筛选适合于大量培养的优良藻种提供了依据。     

气象因素对心力衰竭的影响

目的:相关性分析近两年上海的气象因素及心力衰竭发病患者之间的关系,探讨影响心力衰竭高发的天气气候条件,为开展疾病预防和干预提供了理论和实践依据。方法:前瞻性统计2011年1月-2012年12月上海市胸科医院急诊的心力衰竭人数,并与同期气温,气压,湿度等气象资料进行相关性分析。结果:温度、气压、湿度对心衰的发生具有显著的影响。心力衰竭数与平均气温、最高气温、最低气温呈显著负相关,而与日平均气压呈显著正相关。前期相对湿度变化与心衰数显著相关。温度对心力衰竭数的影响具有滞后效应。结论:气温、气压及湿度与心力衰竭发病有明显相关,揭示了急性呼吸系统感染高发的天气气候条件,为开展疾病预防和干预提供了理论和实践依据。     

Growth and Colony Formation of the Phytoflagellate Prymnesium parvum Carter on Solid Media

SYNOPSIS. Colonies of Prymnesium parvum from single cells were obtained in an atmosphere of high humidity, in soft-agar overlayers on solid medium. Light intensity above 60 foot candles at the surface of the agar plates inhibited colonial development. Such colonies were used to obtain pure lines of the phytoflagellate.
A method for direct demonstration of hemolysin formation on agar plates was developed. The occurrence of hemolytic activity by P. parvum colonies was light-dependent.     

影响翅果油树种子萌发及幼苗生长的几种因素的比较研究

通过对翅果油树种子进行了适温处理、变温处理、激光处理、超声波处理、H₂O₂处理、PEG处理,对发芽实验进行研究,然后测定发芽率、发芽势、萌发指数、根长、芽长、株高、茎粗以及叶绿素含量。萌发实验的结果表明, 变温预处理和PEG预处理能最有效地解除抑制物对种子萌发的影响,对种子进行变温处理和PEG处理是解除种子休眠和提高萌发率的有效途径。本实验为促进濒危植物翅果油树的种群繁衍以及对翅果油树的可持续利用提供一定的科学依据。     

Effects of ambient PO2 and temperature on oxygen uptake in Nautilus pompilius

This study employs closed-circuit respirometry to evaluate the effect of declining ambient oxygen partial pressure (PO₂) and temperature on mass specific rates of oxygen uptake (V˙O₂) in Nautilus pompilius. At all temperatures investigated (11, 16, and 21 °C), V˙O₂ is relatively constant at high PO₂ (oxyregulation) but declines sharply at low PO₂ (oxyconformation). The critical PO₂ below which oxyconformation begins (P _c) is temperature dependent, higher at 21 °C (49 mmHg) than at 11 °C or 16 °C (21.7 mmHg and 30.8 mmHg respectively). In resting, post-absorptive animals, steady-state resting V˙O₂ increases significantly with temperature resulting in a Q₁₀ value of approximately 2.5. The metabolic strategy of N. pompilius appears well suited to its lifestyle, providing sufficient metabolic scope for its extensive daily vertical migrations, but allowing for metabolic suppression when PO₂ falls too low. The combination of low temperatures and low PO₂ may suppress metabolic rate 16-fold (assuming negligible contributions from anaerobic metabolism and internal O₂ stores), enhancing hypoxia tolerance. Accepted: 20 January 2000     

温度对橄榄蛏蚌耗氧和氨氮排泄的影响

The respiration metabolismand excretion of marinebivalves were studied by different researchers[1—6].Themetabolic rate of bivalves is influenced by a number ofvariables,includingtemperature,body size,oxygen ten-sion,food concentration,reproductive state,activityleveland physiological condition.The excreted metabolites ofbivalves include ammonia,urea,uric acid and others,with ammonia comprising70%of the total excretion.Solenaia oleivorais a proper freshwater bivalve in China.For the consumer it has the follo...