麦洼牦牛乳腺上皮细胞系的建立及其生物学特性

为建立麦洼牦牛乳腺上皮细胞的体外培养方法,采用组织块贴壁法及Ⅰ型胶原酶消化法,从麦洼牦牛乳腺组织中成功分离并获得纯化的乳腺上皮细胞,并利用细胞角蛋白18的免疫荧光染色对乳腺上皮细胞进行鉴定。通过细胞生长曲线、群体倍增时间、细胞接种存活率、细胞活力等生物学特性的检测,建立并鉴定麦洼牦牛乳腺上皮细胞系。结果显示,乳腺细胞形态良好,细胞接种存活率达91%,群体倍增时间26.97h,细胞生长曲线呈典型"S"型,细胞传至25代以上仍保持旺盛的增殖活力。可见,通过对细胞传代及反复冻存和复苏已成功建立麦洼牦牛乳腺上皮细胞系,获得大量的乳腺上皮细胞,使麦洼牦牛物种在细胞水平上得以保存,为建立牦牛乳腺生物反应器及泌乳调控机制研究提供基础。     

秋水仙碱诱导银杏花粉染色体加倍的研究

该试验以北京地区银杏雄花芽为材料,探索利用不同浓度的秋水仙碱人工诱导花粉染色体加倍的方法和可能性.用浓度为0.1%、0.2%、0.3%、0.4%、0.5%、0.6%的秋水仙碱溶液,分别对水培和自然非离体条件下诱导雄配子染色体加倍的方法进行了探索,比较了两种情况下不同处理浓度和处理方法诱导效果的差异.结果表明:自然非离体状态下棉浸法处理得到了大花粉,但得率较低,最高为7%,若进一步加大药液处理浓度或适当延长处理时间,将有可能较大幅度提高大花粉得率;水培条件下的棉浸法和注射法处理均未得到大花粉,其原因和适宜的诱导方法还有待进一步探索.以正常花粉作对照,用激光扫描共聚焦显微镜(简称LSCM)对经秋水仙碱处理诱导出的大花粉进行了DNA含量的定量测定,初步确认该试验诱导的大花粉是一种二倍体花粉.      

不同加倍方法处理玉米单倍体的加倍效果研究

以不同基因型玉米材料诱导产生的单倍体幼苗为材料,在2~3叶期用0.06%浓度的秋水仙素配以2.0%的二甲基亚砜（DMSO）通过浸根、滴注心叶和针刺生长点3种不同的方法对单倍体进行人工化学加倍,并对不同加倍方法的加倍率和致死率进行方差分析和多重比较。结果表明,针刺生长点的方法加倍效果最好。平均加倍率为23%,结实率为21.4%,致死率为16.3%。与其他2种化学加倍方法比较,不仅加倍率显著提高,而且所用的秋水仙素剂量更低。     

Inhibition of endothelial regeneration by type-beta transforming growth factor from platelets

Damage to the vessel wall is a signal for endothelial migration and replication and for platelet release at the site of injury. Addition of transforming growth factor-beta (TGF-beta) purified from platelets to growing aortic endothelial cells inhibited [3H]thymidine incorporation in a concentration-dependent manner. A transient inhibition of DNA synthesis was also observed in response to wounding; cell migration and replication are inhibited during the first 24 hours after wounding. By 48 hours after wounding both TGF-beta-treated and -untreated cultures showed similar responses. Flow microfluorimetric analysis of cell cycle distribution indicated that after 24 hours of exposure to TGF-beta the cells were blocked from entering S phase, and the fraction of cells in G1 was increased. The inhibition of the initiation of regeneration by TGF-beta could allow time for recruitment of smooth muscle cells into the site of injury by other platelet components.     

Marc-145细胞低血清适应培养传代稳定性和无血清驯化研究

通过驯化建立无血清悬浮培养型Marc-145细胞系,分析驯化传代对细胞特性的影响,为大规模生产疫苗用的宿主细胞提供基础。采用缓降培养液中血清浓度的方法,连续传代培养50代,并用无血清培养基进行悬浮培养驯化,对传代和驯化过程中的P10、P20、P30、P40和P50代细胞的形态、倍增时间、生长曲线、染色体和对蓝耳病病毒敏感性等特性进行对比分析。研究发现,传代过程中Marc-145细胞均呈扁平多边形上皮样生长,P10代细胞倍增时间为34.15 h,P50代细胞倍增时间为36.16 h,P50代细胞倍增时间稍有延长但差异不显著。P10、P20、P30、P40和P50代的Marc-145细胞生长曲线均呈“S”形;P10代和P50代细胞染色体众数都集中在88~90条。接种病毒后,P10、P20、P30、P40和P50细胞TCID₅₀间无显著差异,表明细胞传50代后,细胞特性没有发生明显变化。P50细胞进行无血清悬浮培养48 h后,细胞密度可达到4.14×10⁶ mL^-1。     

用组织培养法进行马铃薯体细胞染色体的加倍

用马铃薯普通栽培种的２个一单倍体、２个纯合二倍体和５个双单倍体材料，以半叶、茎、叶柄作为外植体通过先诱导愈伤组织，再分化苗的两步法进行了染色体的加倍实验。结果表明：用组织培养二步法可以获得较高的加倍率；加倍率高达９３．３３％。成愈率、苗分化率和加倍率均以半叶为最高，其次为茎段和叶柄；苗分化率的高低直接影响加倍率，二者呈显著的正相关（ｒ＝０．７３３６）；适当地延长愈伤组织培养时间有利于提高加倍率，本实验结果以４０ｄ为宜；基因型对成愈率、苗分化率及加倍率均有影响，亲缘关系近的材料这三种频率也相近；倍性低的材料加倍率较高。     

SMEDWI-2 is a PIWI-like protein that regulates planarian stem cells

We have identified two genes, smedwi-1 and smedwi-2, expressed in the dividing adult stem cells (neoblasts) of the planarian Schmidtea mediterranea. Both genes encode proteins that belong to the Argonaute/PIWI protein family and that share highest homology with those proteins defined by Drosophila PIWI. RNA interference (RNAi) of smedwi-2 blocks regeneration, even though neoblasts are present, irradiation-sensitive, and capable of proliferating in response to wounding; smedwi-2(RNAi) neoblast progeny migrate to sites of cell turnover but, unlike normal cells, fail at replacing aged tissue. We suggest that SMEDWI-2 functions within dividing neoblasts to support the generation of cells that promote regeneration and homeostasis.     

噻虫嗪与氯虫苯甲酰胺对非靶标害虫土耳其斯坦叶螨生长发育的影响

【目的】 研究噻虫嗪与氯虫苯甲酰胺两种药剂,对非靶标害虫土耳其斯坦叶螨生长发育及种群参数的影响。【方法】 采用玻片浸渍法、叶碟法,研究土耳其斯坦叶螨对两种药剂的敏感性、生命参数变化。【结果】 经氯虫苯甲酰胺和噻虫嗪处理的土耳其斯坦叶螨成螨死亡率与对照均无明显差异;氯虫苯甲酰胺处理叶螨成螨后,次代除若Ⅱ期外,未成熟期的存活率均高于对照,而噻虫嗪处理时,次代若Ⅱ期的存活率高于对照,卵期、幼螨期和若Ⅰ期的存活率低于对照;经氯虫苯甲酰胺处理后,与对照和噻虫嗪处理相比,产卵前期明显缩短,世代显著延长,且日产卵量显著增多;氯虫苯甲酰胺处理的种群净增殖率R₀ 、内禀增长率r_m 、周限增长率λ比对照和噻虫嗪处理增大,而施用噻虫嗪的叶螨平均世代历期T、种群加倍时间t比对照和施用氯虫苯甲酰胺延长。【结论】 氯虫苯甲酰胺有刺激土耳其斯坦叶螨增长的作用,而噻虫嗪无影响。     

长白落叶松种子活力与染色体核型

于1986年在东北林业大学种苗室进行实验研究。利用人工老化法得到了两批生活力相近而活力不同的长白落叶松(Larix olgensis Hcnry)种子。应用玻板直立发芽并测定其幼苗生长势。同时,对萌发种子根尖细胞做了显微观察及核型分析。实验结果:种子活力发生递减时,幼苗生长势下降,发芽高峰明显推迟。此时,胚根细胞分裂相减少,分裂相细胞粘滞性增强。细胞染色体数目和核型构型没有发生变化,但核型类型由对照的2B变为2 C。可见,种子胚根细胞染色体的变化是种子劣变活力降低的明显指标与特征之一。     

Textural and crystal-fabric anisotropies and the flow of ice masses

Accurate modeling and prediction of glacier response requires a better understanding of the influence of physical anisotropies on creep. To investigate the effects of variations in the degree of preferred crystallographic orientation and ice crystal size on creep, 19 samples of anisotropic glacier ice were deformed in simple shear. Results indicate that the time required for ice samples to reach the minimum strain rate decreases as crystal size increases; an increase in crystal-fabric development from an isotropic fabric to one with a strong single maximum results in an enhancement of the minimum strain rate by a factor of 4; and a doubling of the crystal size results in about a ninefold increase in the minimum strain rate.     

甲基胺草磷加倍玉米单倍体效果的初步研究

以高诱1号诱导的不同基因型玉米单倍体为材料,在3～5叶期用不同浓度的甲基胺草磷溶液进行滴心处理,研究甲基胺草磷加倍单倍体的效果。结果表明：不同浓度的甲基胺草磷溶液的加倍效率存在一定差异,100μmol／L的甲基胺草磷溶液处理的单倍体加倍效率最高;不同遗传背景材料的加倍效果存在差异。     

Robotic observations of dust storm enhancement of carbon biomass in the North Pacific

Two autonomous robotic profiling floats deployed in the subarctic North Pacific on 10 April 2001 provided direct records of carbon biomass variability from surface to 1000 meters below surface at daily and diurnal time scales. Eight months of real-time data documented the marine biological response to natural events, including hydrographic changes, multiple storms, and the April 2001 dust event. High-frequency observations of upper ocean particulate organic carbon variability show a near doubling of biomass in the mixed layer over a 2-week period after the passage of a cloud of Gobi desert dust. The temporal evolution of particulate organic carbon enhancement and an increase in chlorophyll use efficiency after the dust storm suggest a biotic response to a natural iron fertilization by the dust.     

用胶原酶消化法培养德保矮马耳缘组织成纤维细胞初探

 将德保矮马耳缘组织经胶原酶消化法培养成原代细胞，并成功建立起该组织成纤维细胞系。这种细胞贴壁生长，具有密度接触性抑制性质；该方法获得的原代细胞经传代后接种，细胞群体倍增时间（PDT）为35.9 h；细胞染色体众数2n = 64的细胞数占91.3%～92.8%；乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）同工酶分析，没有其它细胞系污染；细菌、真菌、病毒、支原体检测阴性。该系符合ATCC要求的细胞系鉴定项目，成为保护矮马这一国家重要畜禽品种的宝贵遗传资源，并为相关遗传学研究提供了有效的试验材料。同时得出胶原酶消化培养法比组织块培养法在获得原代细胞速度快；组织块培养法的细胞群体倍增时间（PDT）为48 h。     

Tyrosine transaminase induction by dexamethasone in a new rat liver cell line

A cell line derived from normal, adult rat liver has been established; the cells are similar to hepatocytes, as shown by electron microscopy. The addition of dexamethasone to the culture medium induced a three- to sixfold increase in the specific activity of tyrosine alpha-ketoglutarate transaminase; this increase was inhibited by the simultaneous addition of cycloheximide or actinomycin D. The latter, when added to cells given prior treatment with dexamethasone, further enhanced the transaminase activity. Contact-inhibited cells showed a lower response to dexamethasone than exponentially growing cells.     

The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae

Cell division is arrested in many organisms in response to DNA damage. Examinations of the genetic basis for this response in the yeast Saccharomyces cerevisiae indicate that the RAD9 gene product is essential for arrest of cell division induced by DNA damage. Wild-type haploid cells irradiated with x-rays either arrest or delay cell division in the G2 phase of the cell cycle. Irradiated G1 and M phase haploid cells arrest irreversibly in G2 and die, whereas irradiated G2 phase haploid cells delay in G2 for a time proportional to the extent of damage before resuming cell division. In contrast, irradiated rad9 cells in any phase of the cycle do not delay cell division in G2, but continue to divide for several generations and die. However, efficient DNA repair can occur in irradiated rad9 cells if irradiated cells are blocked for several hours in G2 by treatment with a microtubule poison. The RAD9-dependent response detects potentially lethal DNA damage and causes arrest of cells in G2 until such damage is repaired.     

奈乙酸、2,4-D对马铃薯愈伤组织细胞染色体倍性的影响

以３种不同基因型的马铃薯试管苗茎段做材料，用奈乙酸（ＮＡＡ）和２，４ Ｄ的不同浓度进行愈伤组织的诱导并观察染色体的变异情况，试验表明：在同一浓度下２，４ Ｄ诱导愈伤组织的能力优于ＮＡＡ；且在愈伤组织生长过程中，２，４ Ｄ导致导致细胞染色体加倍的能力也明显强于ＮＡＡ；随２，４ Ｄ和ＮＡＡ浓度的增高，细胞染色体的加倍率明显提高，且细胞染色体的混倍现象也逐渐增高。不同的激素及不同激素浓度对愈伤组织的诱导率和细胞加倍率不同；随生长素浓度的增高，染色体的加倍率明显提高且细胞染色体的混倍现象也逐渐增高。     

花楸种子变温催芽处理的研究

花楸是一种重要的观赏树种,具有重要的经济价值。花楸种子具有生理休眠特性,导致催芽时间长,发芽率低,幼芽长势弱,通常种子处理需要70～90 d,发芽率仅在40%～50%。通过试验采用两段或三段的变温处理花楸种子,催芽时间缩短至60～65 d,发芽率提高到82%～84%。     

二斑叶螨实验种群生命表的组建与分析

在室温25℃、光照16 h、相对湿度20％条件下,组建了二斑叶螨实验种群生命表。结果表明,实验种群的内禀增长力rm为0．1817,周限增长率λ为1．1993d-1,净增殖率R0为25．863 2,平均世代周期T为 17．902 9 d,种群增长1倍需要3．814 5 d。     

雪松年内径向生长节律研究

研究树木年内径向生长规律(intra-annual radial growth)可以为树木生长对气候变化响应研究提供高分辨率基础数据。以雪松为研究对象,采用微树芯技术通过室内制作石蜡切片的方法观察雪松形成层和木质部细胞在生长季期间的活动过程,运用Gompertz函数模拟雪松径向生长的过程,探讨分析洛阳地区针叶树雪松树木年内径向生长节律,并结合气象数据,分析温度、降水量与年内径向生长的关系。结果表明:1）在生长季期间,形成层和扩大加厚期细胞活动呈现双峰分布,木质部细胞积累过程呈现“S”型曲线。2）通过Gompertz函数模拟雪松生长过程,发现在第162天,径向生长细胞个数和长度均出现最大值,分别为1.15个/d、36.7 μm/d。年内平均累计生长细胞个数达到178.3个,年内平均累计生长量达到5 276.7 μm。3)年内径向生长的进程受日均气温和日均降水量的共同影响,温度在生长季不同时期表现为不同的相关性,降水量在生长季期间表现为正相关。     

Lack of replicative senescence in cultured rat oligodendrocyte precursor cells

Most mammalian somatic cells are thought to have a limited proliferative capacity because they permanently stop dividing after a finite number of divisions in culture, a state termed replicative cell senescence. Here we show that most oligodendrocyte precursor cells purified from postnatal rat optic nerve can proliferate indefinitely in serum-free culture if prevented from differentiating; various cell cycle-inhibitory proteins increase, but the cells do not stop dividing. The cells maintain high telomerase activity and p53- and Rb-dependent cell cycle checkpoint responses, and serum or genotoxic drugs induce them to acquire a senescence-like phenotype. Our findings suggest that some normal rodent precursor cells have an unlimited proliferative capacity if cultured in conditions that avoid both differentiation and the activation of checkpoint responses that arrest the cell cycle.