SD大鼠骨髓间充质干细胞的分离培养和鉴定

目的:探讨体外分离培养SD大鼠骨髓间充质干细胞rMSCs的方法,检测传代细胞的细胞周期,并分析部分细胞表型,对rMSCs进行初步的鉴定。方法:密度梯度离心结合贴壁培养法分离培养rMSCs,传代扩增,倒置显微镜进行细胞形态学观察,免疫组化检测细胞表面抗原,流式细胞仪检测细胞周期。结果:密度梯度离心结合贴壁培养法能有效分离纯化rMSCs,细胞呈均一的成纤维细胞样,细胞周期显示90.16%P1代细胞处于G0/G1期,免疫组化结果为:CD44、CD29阳性、CD34阴性,说明培养的细胞即非造血干细胞,也非成纤维细胞。结论:本实验分离培养的细胞群与间充质干细胞的生物学特性吻合,说明密度梯度离心结合贴壁培养法能有效分离纯化rMSCs,细胞稳定表达CD44、CD29,是实用、可行的方法,所培养的细胞可用于细胞移植等研究。     

Ⅰ型骨质疏松山羊骨髓基质细胞体外成脂、成骨分化的研究

观察体外定向诱导Ⅰ型骨质疏松山羊骨髓基质细胞（BMSCs）分化为脂肪细胞和成骨细胞.建立去卵巢骨质疏松山羊动物模型,全骨髓法分离培养其骨髓基质细胞.第二代细胞分别成脂、成骨诱导培养,油红O染色、碱性磷酸酶染色、茜素红染色判定其分化结果.成脂诱导21 d,油红O染色有脂滴形成;成骨诱导14 d,碱性磷酸酶染色阳性;连续诱导35 d,茜素红染色证实有钙化结节形成.Ⅰ型骨质疏松山羊骨髓基质细胞在体外可以定向诱导分化为脂肪细胞和成骨细胞.     

大鼠骨髓基质干细胞分离培养及向成骨、成脂诱导分化的研究

目的利用细胞原代及传代培养技术将大鼠骨髓基质干细胞诱导分化为成骨细胞和脂肪细胞,为其进一步应用奠定基础.方法全骨髓法分离大鼠骨髓基质干细胞,传代后分别在成骨、成脂诱导条件下继续培养,采用碱性磷酸酶染色、茜素红染色及油红"O"染色观察其成骨及成脂分化结果.结果第2代大鼠骨髓基质干细胞成骨诱导9 d后碱性磷酸酶染色呈阳性细胞,连续诱导14 d后可见矿化结节形成,成脂诱导14 d后可见脂肪细胞形成.结论随着诱导条件的不同,大鼠骨髓基质干细胞在体外可定向分化为成骨细胞和脂肪细胞.     

Ι型骨质疏松山羊骨髓基质细胞体外成脂、成骨分化的研究

观察体外定向诱导Ι型骨质疏松山羊骨髓基质细胞(BMSCs)分化为脂肪细胞和成骨细胞。建立去卵巢骨质疏松山羊动物模型,全骨髓法分离培养其骨髓基质细胞。第二代细胞分别成脂、成骨诱导培养,油红O染色、碱性磷酸酶染色、茜素红染色判定其分化结果。成脂诱导21d,油红O染色有脂滴形成;成骨诱导14d,碱性磷酸酶染色阳性;连续诱导35d,茜素红染色证实有钙化结节形成。Ι型骨质疏松山羊骨髓基质细胞在体外可以定向诱导分化为脂肪细胞和成骨细胞。     

成人骨髓基质细胞的体外培养

本研究从成人的髋骨抽取骨髓液，离心后将骨髓基质细胞悬液接种培养，待细胞贴壁融合后，进行传代扩增培养．并采用流式细胞术进行细胞周期分析，结果表明原代培养和传代培养的细胞均为贴壁、形态不一的骨髓基质细胞，且此体外培养的骨髓基质细胞具有很强的增殖能力．本研究成功地建立了一套完整、简单、可行的体外分离、长期培养扩增人骨髓基质细胞(Human Bone Marrow Stromal Cells hBMSCs)的系统，证明成人骨髓基质细胞能够在体外长期增殖，为人的骨髓基盾细胞的理论研究和临床应用奠定了基础.     

大鼠骨髓基质细胞培养的实验研究

目的探讨体外培养大鼠骨髓基质细胞的可行性,为细胞移植治疗疾病奠定基础.方法采用淋巴细胞分离液(1.077 g/L)离心分离MSCs.加入碱性成纤维生长因子(bFGF)进行培养,并用流式细胞仪进行荧光三标检测鉴定.结果分离后的MSCs在生长因子的作用下出现增殖性生长,经流式细胞仪检测显示CD90,CD106阳性,CD45阴性.结论可在体外培养出较大密度大鼠骨髓基质细胞.     

兔和牛角膜上皮、基质及内皮细胞体外培养和增殖的研究

目的：研究兔和牛角膜上皮、基质及内皮细胞等3种细胞体外培养的生物学特性，探索和改进这3种细胞体外培养的方法，为组织工程角膜奠定基础。方法：采用消化法分离培养兔和牛角膜上皮、内皮和基质细胞，观察细胞贴壁生长情况，同时对生长良好的原代细胞进行消化传代，进一步观察传代细胞的形态和生长情况。结果：角膜上皮、内皮和基质细胞均在培养48～72h贴壁生长，培养6～9d能形成良好单层，细胞形态典型，进行多次传代后细胞仍维持原有形态和功能，获得的细胞能进行长期培养。结论：为兔和牛角膜上皮、内皮和基质细胞体外培养，提供了简单高效经济的方法。     

大鼠骨髓基质细胞体外定向诱导成骨

将大鼠骨髓单细胞悬液静置培养36h，利用骨髓基质细胞贴壁能力强的特点对其进行纯化和扩增培养。采用爬片培养、HE染色、组化染色以及碱性磷酸酶活性和钙含量测定等手段研究培养骨髓基质细胞的形态、分化和分泌基质情况。结果表明，非诱导培养条件下骨髓基质细胞呈梭形，部分传代细胞中可观察到脂肪细胞和肌细胞。经成骨性诱导培养后，骨髓基质细胞发生明显的形态学变化，碱性磷酸酶活性上升，钙含量增加，最终形成典型的矿化结节。提示培养大鼠骨髓基质细胞具有分化成脂肪细胞和肌细胞的能力，但其分化成骨的潜能最为强大。本实验诱导骨髓基质细胞分化为成骨细胞的模式有可能适用于骨组织工程研究。     

精原干细胞体外培养的方法改进

探讨精原干细胞体外培养优化方法,为进一步探讨其生物特征、生殖干细胞移植及生殖损伤及药物保护等研究提供实验基础.制备骨髓基质饲养层,将生后5 d~7 d雄性小鼠生精细胞接种在饲养层上,并于IMDM培养基及37 ℃下共培养,在倒置显微镜下观察细胞生长行为,并对培养后的细胞进行组织学、细胞化学、免疫组化、遗传学鉴定.精原干细胞能在以骨髓基质细胞饲养层上进行增殖和生长,且增殖后的表现出呈簇、团状生长,细胞间可表现出明显的胞质间桥,细胞核大,核/质比高,碱性磷酸酶及C-KIT受体阳性,染色体核型为20对(40条)等精原干细胞的生物学行为特征.原代培养的骨髓基质细胞作饲养层具有取材及制作简便等优点,且能在不添加外源性生长因子的前提下能通过自分泌足够的细胞生长因子,满足精原干细胞体外增殖和抑制分化的需要,是精原干细胞培养的一种良好的饲养层.IMDM培养基、血清及适宜的环境温度是精原干细胞体外培养的重要条件.     

体外诱导人骨髓间充质干细胞向胰岛β样细胞分化的研究

目的：探讨体外诱导人骨髓间充质干细胞（ＭＳＣs）向胰岛β样细胞分化。方法：在无菌条件下从正常成人骨髓中分离间充质干细胞,体外培养传3代后用表皮生长因子（epidermal growth factor,EGF）、β-巯基乙醇和高糖培养基诱导MSCs向胰岛β样细胞分化。观察MSCs在诱导前后的形态变化;用胰岛素免疫细胞化学染色检测胰岛素的表达;用双硫腙染色鉴定胰岛β样细胞。结果：未经诱导的MSCs在培养体系中呈贴壁生长,长梭形,经诱导分化后,细胞逐渐变圆,并聚集成团;胰岛素免疫细胞化学表明细胞团内的细胞呈胰岛素染色强阳性反应：双硫腙染色阳性。结论：人骨髓间充质干细胞可在体外被定向诱导分化为胰岛β样细胞。     

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.     

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.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

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。     

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.     

海南岛地体及其毗邻陆缘晚中生代—新生代古地磁研究和构造演化

本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

Jane Eyre——A Feminist In The 19th Century

In the 19th century the society was controlled by men, and women were just appendants of them, they had not any rights and freedom. But Jane was an exception, she showed some characteristics of early feminist. Jane showed her characteristics of feminism in three aspects: rebellion, equality, and independence. These characteristics were helpful to her success, and feminism is the only way out for women of that time.     

《Chinese Science Bulletin》SUBJECT INDEX TO VOLUME 59(2014)

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