小鼠胚胎GMP玻璃化超低温保存研究

本实验研究了GMP法(glass micropipette)冷冻保存小鼠胚胎的主要技术环节,并与目前常用的OPS(open pulled straws)法比较了冷冻后囊胚率.实验表明,EFS30(87.71%)保护作用比EFS40(81.67%)好,但无显著差异(P＞0.05);10%EG(91.37%)预处理优于5%EG(85.93%)、20%EG(88.70%),但差异不显著(P＞0.05);玻璃化时间10～30 s(89.47%、86.79%)显著优于(P＜0.05)60 s(64.00%).GMP法与OPS解冻后囊胚率分别为94.02%和92.06%,差异不显著(P＞0.05),但GMP法稍好于OPS法.同时,GMP法操作比OPS法更简便.     

昆明小鼠未成熟卵母细胞OPS和SSV冷冻保存的比较

目的寻找昆明小鼠卵母细胞冷冻保存效率的新途径。方法选用5~6周龄雌性昆明小鼠的未成熟卵母细胞,在不同前处理液(10%EG或10%EG+10%DMSO)中平衡5 min,然后在冷冻液(EFS30、EFS40、EDFS30或EDFS40)中平衡30 s后进行OPS(Open pulled straw)法和SSV(Solid-surface vitrification)法玻璃化冷冻保存。结果小鼠未成熟卵母细胞的OPS法冷冻保存中,用EFS液冷冻的卵母细胞解冻后形态正常率最高为84.4%,成熟率低于16.7%;而在SSV法冷冻保存中,用EFS液冷冻的卵母细胞解冻后形态正常率最高为86.0%,成熟率为46.5%。OPS法和SSV法冷冻小鼠未成熟卵母细胞形态正常率为91.6%和91.4%,与对照组间差异较显著(P>0.01);成熟率为42.9%和59.1%,与对照组差异极显著(P<0.01)。结论多种抗冻保护剂组合使用效果好于单种抗冻保护剂;EDFS冷冻液冷冻保存效果好于EFS,尤其是EDFS30;OPS法可以有效地冷冻保存小鼠未成熟卵母细胞,且新型玻璃化冷冻法SSV法冷冻保存效果优于OPS法。     

实验用小型猪胎儿成纤维细胞和骨髓间充质细胞为核供体生产转基因克隆胚胎

目的比较不同类型体细胞对生产转基因克隆胚胎效率的影响。方法利用脂质体介导的方法将质粒pEGFP-N1转染到五指山小型猪胎儿成纤维细胞和骨髓间充质细胞,经过G418筛选后均获得了阳性细胞株。然后分别以两种类型转基因细胞以及未转基因细胞为核供体进行体细胞核移植,比较不同类型供体细胞克隆胚胎的囊胚发育率。结果胎儿成纤维细胞和骨髓间充质细胞克隆胚的囊胚发育率差异不显著(P>0.05,8.3%vs.7.1%);转基因胎儿成纤维细胞(9.6%)和转基因骨髓间充质细胞(9.9%)克隆胚胎的囊胚发育率差异不显著(P>0.05,9.6%vs.9.9%);每一种类型供体细胞转基因与否对克隆胚的囊胚发育率无影响(P>0.05)。结论通过体细胞核移植技术,小型猪骨髓间充质细胞与胎儿成纤维细胞均可有效地生产转基因囊胚。     

小型猪超数排卵与胚胎回收方法的研究

目的本试验对小型猪进行超数排卵和早期胚胎回收技术进行试验研究。研究结果如下:PG PMSG HCG法对断奶母猪的同期发情率达100%,优于其对非哺乳期空怀母猪的72.73%的同期发情率,但二者平均排卵数显著性差异(P<0.05);屠宰冲胚和手术法冲胚,胚胎回收率分别为84.29%和76.92%,回收率差异显著(P<0.05),但头均回收胚数无显著性差异(P>0.05)。     

阴离子盐对人工瘤胃体外发酵的影响

应用体外产气法测定阴离子盐对人工瘤胃体外发酵产气量、pH值、干物质消失率及NH3-N浓度的影响。试验分为3个组,对照组和2个试验组饲粮干物质中自制阴离子盐分别占0%、1.87%、3%。结果显示,阴离子盐对人工瘤胃的最大产气量没有影响,但显著降低了产气速率(P<0.05);与对照组相比,2个试验组干物质消失率有所降低,但差异不显著(P>0.05)。发酵24h后,试验组pH值比对照组有所提高,但差异不显著(P>0.05);试验1组NH3-N浓度显著高于试验2组(P<0.05),但对照组和试验组间差异不显著(P>0.05)。     

不同冷冻方法和解冻液对猪精液冷冻效率的影响

采用三种不同的冷冻方法冷冻猪的新鲜精液,再分别用不同的解冻液在相同条件下同时解冻,对比解冻后精子的活力和顶体完整率,以优化猪精液冷冻方法.实验一:随机选取三头健康的有稳定受精能力的优良种公猪,采集新鲜精液,分别用干冰颗粒、液氮颗粒和液氮细管三种方法冷冻,用同样的方法解冻,对比解冻后精子的活力和顶体完整率;实验二:用不同的解冻液解冻经干冰颗粒法冷冻的精液,对比不同解冻液对精子活力的影响;实验三:以孤雌激活和鲜精受精为对照,验证经实验一和实验二选择出来的最优冷冻方法和解冻液处理后的精子的体外受精能力.实验一显示,干冰颗粒冷冻法冷冻的猪精液解冻后精子活力(0.23)略高于液氮颗粒法的精子活力(0.20),而二者均显著高于细管冻精法的精子活力(0.08)(P<0.05),液氮颗粒法的顶体完整率(54.7%)和细管法的顶体完整率(54.2%)无显著差异,但二者均显著低于干冰颗粒法的顶体完整率(58.0%)(P<0.05).实验二显示,Glucose-EDTA解冻液解冻后,精子活力(0.22)显著高于DPBS-BSA解冻液解冻后的精子活力(0.13)(P<0.05).实验三:经干冰颗粒法冷冻、Glucose-EDTA解冻液解冻的精液,体外受精后的囊胚发育率(17.6%)与鲜精体外受精囊胚率(17.9%)无显著差异,但二者均显著低于孤雌激活对照组的囊胚率(36.6%)(P<0.05).由此可知:干冰颗粒法冷冻结合Glucose-EDTA解冻液解冻是较为理想猪精液冷冻保存方法,经该方法冷冻、解冻的猪精液,体外受精后的胚胎发育率与鲜精没有显著差异.     

细胞松弛素B、温度、离心处理对猪卵母细胞OPS玻璃化冷冻的影响

主要探讨使用CB预处理和不同温度冷冻保护剂对猪卵母细OPS玻璃化冷冻保存的影响,并对使用不同温度冷冻保护剂下,探讨离心猪卵母细胞后的OPS冷冻、解冻的效果.结果表明,体外成熟培养12h后,将经7.5μg/mL细胞松弛素B(CB)处理的猪卵母细胞进行OPS玻璃化冷冻保存.解冻后继续体外成熟培养,猪卵母细胞的形态正常率和成熟率与对照组无显著差异(P>0.05),但是与对照组相比,经冷冻前经CB预处理的卵母细胞其体外受精胚胎的卵裂率、桑葚胚和囊胚率显著提高(P<0.05).此外,使用预平衡至39℃的冷冻保护剂能够达到较好的解冻效果,卵母细胞成熟率较高,但离心并没有促进冷冻后猪卵母细胞体外受精胚胎发育率.     

克隆绵羊遇到的问题及其分析

本研究以体外成熟不同时间的绵羊卵母细胞为胞质受体,对去核、融合、不同激活方法、卵裂率、桑/囊胚率和克隆胚受胎率等作了研究与分析.观察妊娠受体母羊的状态、胎儿体内发育情况,分析影响胎儿发育的原因.结果显示,成熟培养18h、22h、24h卵母细胞的盲吸去核率无显著性差异(84.9%,86.6%,84.3%,p>0.05),但显著高于26h组的去核率(71.7%).离子酶素联合cycloheximide(CHX)对重构胚进行激活,卵裂率显著高于A23187联合6-DMAP的激活方法(84.6%vs 63.2%,p<0.05),但二者处理对桑/囊胚率发育无显著差异(25.5%vs18.7%,p>0.05).作为胞质受体,成熟20h的卵母细胞与成熟24h和26h的卵母细胞相比,不利于重构胚的体外发育,桑/囊胚率差异显著(12.3%vs 25.3%,23.4%,p<0.05).分别以美丽奴、杜泊肉羊的体细胞为供体细胞进行克隆生产,90天以后的妊娠率为10%-17%,发育到期率为10%以上.出生羔羊的体重普遍较大.     

克隆牛过程中电融合条件对附植前胚胎发育的影响

实验比较了克隆牛过程中电融合条件的改善对附植前胚胎发育的影响. 电融合强度为140 V/mm,10 μs/次时的效率要较100 V/mm,20 μs/次和180 V/mm,10 μs/次时高,可以降低融合后重构胚死亡率,提高克隆牛囊胚发育率. 施加直流电脉冲前先给予5 V交流电场预处理数秒钟,然后在电极之间通以两个直流脉冲(140 V/mm, 10 μs/次,间隔0.1 s)与不用交流电预处理对融合率和重构胚的发育率无显著差异. 另外实验比较了甘露醇和山梨醇两种融合液对融合及重构胚发育的影响,结果表明囊胚发育率差异不显著,但山梨醇较甘露醇融合液更容易操作. 实验生出的4头克隆牛存活至今.     

瘦素对猪卵母细胞体外成熟及克隆猪妊娠率的影响

体细胞克隆猪在人类医学、基础科学研究和畜牧业生产方面均具有很大的应用潜力.为了提高体细胞克隆猪的效率,首先比较了两种基础成熟液NCSU-23和TCM199的体外成熟效果,然后在TCM199培养液的基础上,较系统地研究了添加不同浓度的瘦素对猪卵母细胞成熟、孤雌激活胚胎和克隆胚胎的体外发育以及克隆胚胎体内发育的影响.结果表明:成熟液中添加100ng/mL或200ng/mL瘦素对猪卵母细胞的核成熟没有显著影响(P>0.05);对孤雌激活卵裂率和克隆胚胎卵裂率也没有显著影响(P>0.05);但却显著提高了孤雌激活胚胎的囊胚率和囊胚细胞数,并且显著提高了克隆胚胎的囊胚率;当添加量为100ng/mL时,克隆囊胚的细胞数显著升高.另外,研究表明,瘦素很可能具有提高克隆胚胎移植妊娠率和妊娠到期率的作用.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

Quantitative trait loci （QTLs） for quality traits related to protein and starch in wheat

Quality traits in wheat （Triticum aestirum L.） were studied by quantitative trait locus （QTL） analysis in a recombinant inbred line （RIL） population, a set of 131 lines derived from Chuan 35050 × Shannong 483 cross （ChSh）. Grains from RILs were assayed for 21 quality traits related to protein and starch. A total of 35 putative QTLs for 19 traits with a single QTL explaining 7.99-40.52% of phenotypic variations were detected on 10 chromosomes, 1D, 2A, 2D, 3B, 3D, 5A, 6A, 6B, 6D, and 7B. The additive effects of 30 QTLs were positive, contributed by Chuan 35050, the remaining 5 QTLs were negative with the additive effect contributed by Shannong 483. For protein traits, 15 QTLs were obtained and most of them were located on chromosomes 1 D, 3B and 6D, while 20 QTLs for starch traits were detected and most of them were located on chromosomes 3D, 6B and 7B. Only 7 QTLs for protein and starch traits were co-located in three regions on chromosomes 1D, 2A and 2D. These protein and starch trait QTLs showed a distinct distribution pattern in certain regions and chromosomes. Twenty-two QTLs were clustered in 6 regions of 5 chromosomes. Two QTL clusters for protein traits were located on chromosomes 1D and 3B, respectively, three clusters for starch traits on chromosomes 3D, 6B and 7B, and one cluster including protein and starch traits on chromosome 1D.     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.