异烟肼对斑马鱼胚胎发育量-毒关系的研究

研究了一线抗结核药物异烟肼对斑马鱼发育的量-毒关系。用不同浓度的异烟肼分别处理发育至2 hpf(hour post fertilization)的斑马鱼胚胎,观察胚胎和幼鱼的死亡率、孵化率、畸形率、体长及形态学变化。急性毒性实验结果表明异烟肼对斑马鱼有明显的发育毒性,能导致斑马鱼发育迟缓和畸变。与对照组相比,低剂量（小于6 mmol/L）异烟肼对斑马鱼无发育毒性,高剂量（≥32 mmol/L）异烟肼显著抑制胚胎的孵化率,并有明显的胚胎致死效应和致畸效应。斑马鱼死亡率和畸形率均随给药剂量的增加和给药时间的延长而升高。本实验条件下,异烟肼的最大无毒性效应浓度为2 mmol/L。     

汞和铅的单一和复合作用对斑马鱼胚胎发育的毒性效应研究

基于斑马鱼胚胎技术,研究了汞(Hg~(2+))和铅(Pb~(2+))对斑马鱼胚胎发育的单一及联合毒性,主要观察和比较了96 h斑马鱼胚胎的死亡率、孵化率和畸形率,并采用相加指数法分析比较了Hg~(2+)和Pb~(2+)在96 h对斑马鱼胚胎发育的效应类型.结果表明,斑马鱼胚胎对Hg~(2+)和Pb~(2+)均表现出显著的浓度依赖性,造成胚胎孵化率下降,发育畸形甚至死亡;Hg~(2+)的毒性作用高于Pb~(2+),Hg~(2+)和Pb~(2+)的96 h-LC50分别为0. 081 mg/L和113. 978 mg/L; Hg~(2+)和Pb~(2+)组合在96 h表现为拮抗作用.     

石油烃分散液对马粪海胆浮游幼虫的急性毒性效应

为研究石油烃污染对海胆胚胎的急性毒性效应,采用胚胎发育毒性实验方法,分6个质量浓度组(1.0、2.0、4.0、8.0、16.0、32.0 mg/L)分别进行0#柴油和船用燃料油分散液对马粪海胆浮游幼虫发育的急性毒性实验.结果显示,两种油品分散液对胚胎发育至二腕幼虫和四腕幼虫阶段有明显的毒性作用.随着受试液质量浓度的增加,幼虫的体长变短,畸形率及死亡率显著增大.船用燃料油分散液48 h和72h的半数效应浓度(EC50)分别为4.57、2.71 mg/L;0#柴油分散液48 h和72 h的EC50分别为3.39、1.87 mg/L,故0#柴油的毒性大于船用燃料油.     

3种重金属对斑马鱼的联合毒性及迷迭香酸的保护作用研究

对氯化镉、硝酸铅、硫酸铜3种重金属单一以及联合情况下对早期斑马鱼胚胎发育毒性的影响进行考察,并采用不同浓度的迷迭香酸进行干预,同时观察96 hpf胚胎的死亡率和孵化率.Cd-Pb、Cd-Cu、Pb-Cu、Cd-Pb-Cu对斑马鱼胚胎的毒性作用类型分别为协同作用、协同作用、拮抗作用、协同作用.60μmol/L RA对斑马鱼胚胎死亡、幼鱼畸形及胚胎孵化率无明显影响.Cd~(2+)-Pb~(2+)-Cu~(2+)联合处理,胚胎死亡率明显上升,60μmol/L RA与Cd~(2+)-Pb~(2+)-Cu~(2+)联合处理,胚胎死亡率降低约20%(P<0.01);胚胎凋亡细胞数目明显减少,Bcl-2/Bax的基因表达水平明显上调(P<0.01),Caspase 3基因表达下调(P<0.05).表明RA对Cd~(2+)-Pb~(2+)-Cu~(2+)的联合毒性具有保护作用,其机制可能与上调Bcl-2/Bax,降低Caspase 3基因表达,减少胚胎细胞凋亡有关.     

利用人胚胎干细胞评价反式维甲酸的胚胎发育毒性

目前,药品安全、食品安全和环境污染等问题受到国人的广泛关注,也受到政府的特别重视。医用化学品、食品添加剂和环境污染物等通过各种途径进入体内,可能产生胚胎发育毒性;因此,建立新的高通量、高灵敏的胚胎发育毒性检测和评价十分重要。研究尝试利用全反式维甲酸(all-trans retinoic acid,RA)对人胚胎干细胞H9的细胞毒性和分化抑制,评价RA的胚胎发育毒性。通过CCK8检测不同浓度下RA对人胚胎干细胞H9和鼠胚胎成纤维细胞3T3的细胞存活百分数和半数增殖抑制浓度(50%inhibitory concentration,IC50),发现处理5 d和10 d,RA对人胚胎干细胞H9的IC50分别是9.23μg/mL和7.20μg/mL,明显低于3T3细胞。通过实时定量PCR检测不同RA浓度下,自然分化20 d的细胞中Nkx2.5、α-MHC、ACTC1和TNNT2基因的表达,发现0.3μg/mL及其以上浓度RA显著抑制H9细胞向心肌细胞分化。RA对H9细胞α-MHC、TNNT2和ACTC1的半数抑制分化浓度(ID50),分别为0.16、0.07和0.05μg/mL,ACTC1和TNNT2的ID50明显低于α-MHC的,提示ACTC1和TNNT2可能更适合作为人胚胎干细胞分化抑制的评价的指标。     

中国林蛙胚胎后期发育观察

首次对中国林蛙在华南地区的胚胎后期发育进行观察研究.结果表明:华南地区的环境条件基本适合中国林蛙的胚胎后期发育,较高的温度加快了中国林蛙的胚胎发育进程,但对其生长无明显影响.     

卫河水对太行隆肛蛙蝌蚪红细胞核异常的影响

运用遗传毒理学的方法,研究环磷酰胺和卫河水对太行隆肛蛙蝌蚪红细胞核异常的影响.结果表明:经10 mg.L-1、20 mg.L-1、40 mg.L-1环磷酰胺溶液和10%、30%、50%体积浓度的卫河水处理4、8、12和16 d,太行隆肛蛙蝌蚪红细胞的微核率和总核异常率明显升高;20 mg.L-1环磷酰胺溶液可作为阳性对照;每一处理浓度下,红细胞微核率、总核异常率均在第12 d达到高峰.说明,太行隆肛蛙蝌蚪红细胞微核和核异常实验对监测污染水体中的遗传毒性物质敏感可行;卫河水污染严重,含有较强的遗传毒性物质,应加快对卫河水的治理,以防止其对地下水源的污染.     

邻苯二甲酸酯对斑马鱼胚胎发育的联合毒性

通过48 h单一毒性试验和正交试验,研究了邻苯二甲酸二甲酯(DMP)、邻苯二甲酸二乙酯(DEP)、邻苯二甲酸二正丁酯(DBP)和邻苯二甲酸二异辛酯(DEHP)4种邻苯二甲酸酯类化合物暴露对斑马鱼胚胎发育的毒性效应。结果表明:这些化合物对斑马鱼胚胎发育有明显的抑制作用,可以造成胚胎发育畸形甚至死亡,具有特定最敏感的毒理学终点;4种邻苯二甲酸酯(PAEs)单独作用时毒性大小顺序依次为DBPDEHPDEPDMP;在4种PAEs对斑马鱼胚胎发育的混合暴露正交试验中,DBP和DEHP存在交互作用,对其致死毒性和发育毒性影响最大;DBP虽然对斑马鱼胚胎的致死毒性和发育毒性影响不明显,但DBP可导致胚胎发育的严重畸形。     

不同LED光谱对斑马鱼胚胎发育的影响

【目的】考察不同LED光谱对斑马鱼胚胎发育的影响。【方法】将斑马鱼胚胎随机平均分为4组分别置于白光、蓝光、绿光和红光共4种LED光谱环境中，每日光照时间段为7:00—21:00，持续7 d。在此期间每12 h统计胚胎孵化率和胚胎或仔鱼死亡率，记录胚胎在第48 h的心率、仔鱼在第168 h的畸形率及体长，并拍摄胚胎在第4 h、第12 h、第24 h和第48 h的发育情况。【结果】1) 与白光组相比，蓝光组和红光组在第60 h以及红光组在第84 h的胚胎累计孵化率有统计学意义上的下降(p<0.05)，而第96 h后各组胚胎累计孵化率无统计学意义上的差异，但绿光组的该项指标最低。2) 各组斑马鱼胚胎或仔鱼累计死亡率在各统计时间点均无统计学意义上的差异，但胚胎完全孵化后蓝光组的该项指标最低。3) 各组斑马鱼胚胎心率在第48 h无统计学意义上的差异；4) 各组斑马鱼仔鱼的体长和畸形率无统计学意义上的差异，但红光组仔鱼畸形率最高。【结论】最适宜斑马鱼胚胎发育的LED光谱为白光，红光和蓝光会延缓该鱼种的胚胎发育；红光和绿光会导致斑马鱼胚胎或仔鱼死亡率有一定程度上升，对该鱼种的胚胎发育较为不利。     

悬浮物对斑马鱼胚胎发育的影响

为探究水中悬浮物含量对斑马鱼(Danio rerio)胚胎发育的影响，挑选发育正常的斑马鱼胚胎分为CK组、T1组、T2组和T3组，分别在悬浮物质量浓度为0、10、100和1 000 mg·L -1的水体中培养168 h，每12 h记录胚胎孵化率和胚胎或仔鱼的死亡率，在实验进行72 h后统计仔鱼心率，在实验进行168 h后统计仔鱼畸形率，并于实验进行4、12、24和48 h后拍照观察胚胎的发育情况。结果显示：1) 与CK组相比，T1组、T2组和T3组斑马鱼胚胎的累计孵化率在实验进行60 h后有统计学意义上的下降(p<0.05)，且T3组的该项指标在实验进行72 h后直至实验结束均有统计学意义上的下降(p<0.05)；2) 在实验进行120 h后直至实验结束，T3组与CK组相比仔鱼的累计死亡率均有统计学意义上的上升(p<0.05)；3) 在实验进行72 h后，T1组、T2组和T3组与CK组相比仔鱼心率有统计学意义上的下降(p<0.05)；4) T3组与CK组相比仔鱼畸形率有统计学意义上的上升(p<0.05)；5) 与CK组相比，T1组、T2组和T3组的胚胎发育较为迟缓。研究结果提示水体中悬浮物质量浓度过高可导致斑马鱼胚胎发育迟缓、胚胎孵化率和仔鱼心率下降、仔鱼的死亡率和畸形率上升。     

日本沼虾胚胎发育时期消化酶活力和氨基酸含量

采用生物化学方法测定了日本沼虾胚胎发育过程中四种消化酶的比活力和氨基酸的含量.结果显示：卵内可溶性蛋白的含量随着胚胎的发育逐渐下降,但在原肠期的可溶性蛋白含量略高于囊胚期.四种消化酶中类胰蛋白酶、胃蛋白酶和淀粉酶活力在胚胎发育早期和后期较高,中期较低.纤维素酶活力较低且在胚胎发育过程中变化规律不明显.总氨基酸含量在胚胎发育早期下降较快,胚胎发育至后无节幼体期氨基酸含量下降幅度变缓.必需氨基酸中亮氨酸的含量高,非必需氨基酸中谷氨酸的含量最高.单个必需氨基酸含量与必需氨基酸总量的比值（A/E）在整个胚胎发育过程中的变化基本趋于一致.     

Ciliary neurotrophic factor prevents the degeneration of motor neurons after axotomy

The period of natural cell death in the development of rodent motor neurons is followed by a period of sensitivity to axonal injury. In the rat this early postnatal period of vulnerability coincides with that of very low ciliary neurotrophic factor (CNTF) levels in the sciatic nerve before CNTF increases to the high, adult levels. The developmental time course of CNTF expression, its regional tissue distribution and its cytosolic localization (as suggested by its primary structure) favour a role for CNTF as a lesion factor rather than a target-derived neurotrophic molecule like nerve growth factor. Nevertheless CNTF exhibits neurotrophic activity in vitro on different populations of embryonic neurons. To determine whether the vulnerability of motor neurons to axotomy in the early postnatal phase is due to insufficient availability of CNTF, we transected the axons of newborn rat motor neurons and demonstrated that local application of CNTF prevents the degeneration of the corresponding cell bodies.     

空心微珠保温冒口的研制与应用

在铸造生产中采用空心微珠制作保温冒口,在大、中、小型铸钢件上应用,保温效果良好,延长了冒口的凝固时间,不仅消除了铸件的缩孔及缩松等缺陷,而且冒口的补缩效率明显提高,由原来的20％,提高到30％-40％。铸件工艺出品率由原来的60％,提高到70％-73％。大量节约了金属液,节省了人力及能源。     

Gene expression divergence recapitulates the developmental hourglass model

The observation that animal morphology tends to be conserved during the embryonic phylotypic period (a period of maximal similarity between the species within each animal phylum) led to the proposition that embryogenesis diverges more extensively early and late than in the middle, known as the hourglass model. This pattern of conservation is thought to reflect a major constraint on the evolution of animal body plans. Despite a wealth of morphological data confirming that there is often remarkable divergence in the early and late embryos of species from the same phylum, it is not yet known to what extent gene expression evolution, which has a central role in the elaboration of different animal forms, underpins the morphological hourglass pattern. Here we address this question using species-specific microarrays designed from six sequenced Drosophila species separated by up to 40 million years. We quantify divergence at different times during embryogenesis, and show that expression is maximally conserved during the arthropod phylotypic period. By fitting different evolutionary models to each gene, we show that at each time point more than 80% of genes fit best to models incorporating stabilizing selection, and that for genes whose evolutionarily optimal expression level is the same across all species, selective constraint is maximized during the phylotypic period. The genes that conform most to the hourglass pattern are involved in key developmental processes. These results indicate that natural selection acts to conserve patterns of gene expression during mid-embryogenesis, and provide a genome-wide insight into the molecular basis of the hourglass pattern of developmental evolution.     

Evidence that a free-running oscillator drives G1 events in the budding yeast cell cycle.

In yeast and somatic cells, mechanisms ensure cell-cycle events are initiated only when preceding events have been completed. In contrast, interruption of specific cell-cycle processes in early embryonic cells of many organisms does not affect the timing of subsequent events, indicating that cell-cycle events are triggered by a free-running cell-cycle oscillator. Here we present evidence for an independent cell-cycle oscillator in the budding yeast Saccharomyces cerevisiae. We observed periodic activation of events normally restricted to the G1 phase of the cell cycle, in cells lacking mitotic cyclin-dependent kinase activities that are essential for cell-cycle progression. As in embryonic cells, G1 events cycled on schedule, in the absence of S phase or mitosis, with a period similar to the cell-cycle time of wild-type cells. Oscillations of similar periodicity were observed in cells responding to mating pheromone in the absence of G1 cyclin (Cln)- and mitotic cyclin (Clb)-associated kinase activity, indicating that the oscillator may function independently of cyclin-dependent kinase dynamics. We also show that Clb-associated kinase activity is essential for ensuring dependencies by preventing the initiation of new G1 events when cell-cycle progression is delayed.     

环磷酰胺对SD 大鼠生育力与早期胚胎发育毒性阳性模型的建立

摘要: 目的探讨生育力与早期胚胎发育毒性试验中,皮下给予不同剂量的环磷酰胺对SD 大鼠的影响,选择合适的剂量建立阳性模型,积累实验室背景数据。方法取7 ～ 8 周龄健康SD 大鼠176 只,雌雄各半。按体质量随机分为溶媒对照组( Veh) 、环磷酰胺低( L: 10 mg·kg － 1 ) 、中( M: 20 mg·kg － 1 ) 、高( H: 40 mg·kg － 1 ) 剂量组,每组44 只,雌性各半。雄鼠于交配前28 d 开始给药,每周一次,给药期持续整个交配期直至处死; 雌鼠于交配前14 d 开始给药,交配前每周1 次,交配成功后第0 天给药1 次,总共给药3 次。雄鼠及未交配成功雌鼠于交配结束后处死剖检,雌鼠于受孕后第15 d 剖检,统计各剂量组雄鼠生育率; 雌鼠性周期、妊娠率; 孕鼠死亡率和死胎率。结果环磷酰胺低、中、高各剂量组对SD 大鼠生育力与早期胚胎发育均有一定的毒性作用,且随给药剂量增加,毒性作用增强。结论皮下给予SD 大鼠不同剂量的环磷酰胺,高剂量组对大鼠生育力与早期胚胎发育的毒性影响较为明显,故选择此剂量作为本实验室的阳性模型。     

超声波对4种海水鱼类受精卵孵化的影响

采用复合式纵向压电换能器,谐振频率为21．5kHz,谐振阻抗为120Ω,施加电压值为80V的超声波辐射海水鱼类受精卵,研究不同的超声辐射时间和辐射次数对大弹涂鱼、中华乌幅鳢、真鲷和大黄鱼受精卵孵化的影响,结果表明,超声处理大弹涂鱼胚胎发育后期的受精卵,呆提高其孵化率,但对大弹涂鱼和中华乌塘鳢胚胎发育早期受精卵的孵化影响,每次辐射时间为1min、总幅射次数为4次的辐射剂量有利于真鲷受精卵的孵化;每次辐射时间超过1min,孵化率降低,以每次辐射时间为10s或30s处理大黄鱼受精卵,其孵化率分别比对照组高出10％和12％。     

中华蟾蜍指名亚种(Bufo gargarizan gargarizan)早期胚胎发育

2006年1—2月份对中华蟾蜍指名亚种的早期胚胎发育和产卵行为进行观察.结果表明,蟾蜍整个产卵时间为6.18±0.73 h(N=4),其中雌性产卵8.00±1.20次,雄性射精9.25±1.31次.产卵中,雌性产卵的时间间隔为0.75±0.06 h(N=39),抱对的蟾蜍要游到水面上呼吸,两次呼吸间隔为1.39±0.23 h(N=11).雌蟾蜍最后一次产卵到雌雄分开所需时间为0.96±0.19 h(N=7).从受精卵期至鳃盖完成期,胚胎发育可分为23个时期,在6℃～10℃的水温条件下,历时1111.23±21.64 h(N=7).     

Evidence from reversal of handedness in C. elegans embryos for early cell interactions determining cell fates

Many animals with overall bilateral symmetry also exhibit some left-right asymmetries with generally invariant handedness. Therefore, the left-right embryonic axis must have a consistent polarity, whose origins and subsequent effects on development are not understood. Caenorhabditis elegans exhibits such left-right asymmetries at all developmental stages. The embryonic cell lineage is asymmetric as well: although the animal is generally bilaterally symmetric, many of its contralaterally analogous cells arise from different lineages on the two sides of the embryo. I accomplished reversal of embryonic handedness by micromanipulation at the 6-cell stage, which resulted in mirror-image but otherwise normal development into healthy, fertile animals with all the usual left-right asymmetries reversed. This result demonstrates that in the 6-cell embryo the pair of anterior (AB) blastomeres on the right is equivalent to the pair on the left, and that the extensive differences in fates between lineally homologous derivatives of these cells on the two sides of the animal must be dictated by cell interactions, most of which are likely to occur early in embryogenesis.