组织块法培养大鼠肺微血管内皮细胞的方法研究

实验应用SD雄性大鼠的外周肺组织,进行大鼠肺微血管内皮细胞(RPMVEC)的原代培养和传代培养,并从实验动物、培养基、操作条件、剪块方法、植块方法等细节方面对组织块法培养RPMVEC的方法进行探讨改进.同时,对培养细胞进行Ⅷ因子相关抗原免疫细胞化学鉴定,通过光镜和透射电镜观察细胞形态和超微结构,通过MTT观察细胞生长情况.体外培养的原代RPMVEC在光镜下呈多角形或梭形,胞核清晰,呈卵圆形.细胞单层呈典型的铺路石样排列.细胞生长状态良好、抗Ⅷ因子相关抗原染色阳性.透射电镜可见Weibel-palade小体.改进后的组织块培养法获取RPMVEC的成功率高,污染几率低,细胞活力好,纯度高,数量大,同时保有了PMVEC的结构和功能,可用于进一步的实验研究.     

自体细胞来源的组织工程皮肤及其实验研究

利用患者自体来源的皮肤细胞,在经过短时间的体外培养后构建组织工程皮肤,并对其移植后的转归进行动物实验观察.分离培养头皮中的角质形成细胞及创面来源的成纤维细胞,并在体外进行传代扩增.利用三维立体培养系统构建复合细胞膜片,移植于裸鼠背部全层缺损创面,以单层细胞膜片为对照,对其转归进行大体及病理学观察.体外分离培养的皮肤细胞在体外扩增2周后即可用于构建复合细胞膜片,在移植覆盖裸鼠全层皮肤创面后,伤口愈合良好,移植2月后的病理结果显示皮肤结构明显优于单层膜片移植.证明利用患者自体来源的皮肤细胞可快速构建组织工程皮肤,有望成为治疗大面积深度烧伤患者的途径之一.     

牙髓干细胞在修复牙槽骨缺损的组织学观察

为探索牙髓干细胞(DPSCs)在兔牙槽骨缺损再生修复中的成骨效果,为临床上骨缺损的修复提供一定的理论基础。采用从新西兰幼兔前牙及磨牙牙髓组织中分离DPSCs进行体外培养;16只实验新西兰兔随机分为空白组,实验组两组,在动物下颌无牙区牙槽骨人工制备10 mm×4 mm×4 mm骨缺损,其中空白组骨缺损区填入浸有PBS溶液的0.25 g Bio-oss骨粉,实验组加入1×108/L的DPSCs和0.25 g Bio-oss骨粉;于术后第6周同期处死动物,无菌获取牙槽骨组织,制作石蜡切片,HE染色及免疫组化检测骨涎蛋白(BSP)。结果显示:光镜下传代培养的细胞与原代培养的细胞形态基本一样,幼兔牙髓细胞具有体外多次传代增殖能力;术后6周,空白组纤维结缔组织较稀疏,牙槽嵴塌陷,唇舌侧牙槽骨凹陷吸收,HE染色可见少量红细胞,未见明显新骨生成,骨涎蛋白免疫组化成弱阳性表现。实验组牙槽骨缺损区愈合良好,牙槽嵴饱满,可见部分未吸收骨粉颗粒,被新生组织包裹,新生组织与骨缺损周围嵌入良好,上附有部分纤维结缔组织,唇舌侧牙槽骨未见明显凹陷,大部分牙槽嵴宽度及高度与邻牙基本相平,骨缺损区质地较韧,HE染色提示炎性渗出明显吸收,可见新生骨组织,骨缺损区周围有大量成骨细胞,骨小梁较空白组致密,骨涎蛋白免疫组化成强阳性表达,与空白组有明显差异。由此可知,牙髓干细胞有向成骨分化能力,促进牙槽骨缺损的修复。     

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。     

草鱼前体脂肪细胞原代、传代培养及鉴定

建立草鱼(Ctenopharyngodon idellus)前体脂肪细胞的培养体系,体外重现草鱼脂肪细胞的增殖分化过程.实验采用油红0染色法对脂肪细胞进行鉴定;采用倒置显微镜(NIKON)和CCD对细胞进行观察摄像;选用的健康草鱼体重为800～900 g;利用体外组织培养技术在温度为28℃,CO2浓度为5%,血清浓度为20%,pH值为7.0～7.2的条件下,以DMEM/F12培养基对来源于草鱼腹腔的脂肪组织进行原代培养;单层的原代培养细胞达到瓶底面积的70%～80%且汇合时.对细胞进行传代培养.研究发现在本实验条件下,培养48 h后组织块周围有梭形细胞迁出;3 d后细胞数量增多,多为梭形和多边形;培养8 d后大部分细胞融合;细胞内脂滴可被亲脂的油红0着色,证明为脂肪细胞.对融合后的细胞进行传代培养,可传至第4代;传代第3 d可形成致密单层;第8 d胞质内含有大量脂滴.研究初步确立了较为适宜的草鱼前体脂肪细胞离体培养体系.     

中华鳖心肌有限细胞系的建立及其生物学特性

首次对中华鳖的心肌细胞进行了体外原代和传代培养获得成功。并对其细胞形态,生长速率等细胞生物学特性以及温度和ＰＨ值对细胞生长的影响进行了观察研究。     

胚胎干细胞向心肌细胞分化研究进展(综述)

胚胎干细胞(embryonic stem cells,ES)在体外分化培养条件下可以分化出各种组织细胞，其中包括心肌细胞。ES细胞在体外向心肌细胞分化与体内完整胚胎心肌发育过程相符合。该细胞在体外分化过程中顺序表达心肌细胞特有结构蛋白和离子通道，如肌球蛋白轻链和重链、特异性肌动蛋白、电压依赖性Ca^2 通道、K^ 通道等。ES细胞分化来源的心肌细胞具有体内心肌细胞的生理学特点，如产生的动作电位、表现自发性收缩等。因此，ES细胞是研究心肌细胞发育分化机制及鉴定其关键基因的理想模型。     

胎鼠心肌与肝脏细胞的体外生长特性

用M199和DMEM两种培养液分别培养取自14～16日龄胎鼠的心肌、肝脏组织细胞.通过观察细胞生长情况、进行细胞计数和绘制细胞生长曲线,对不同组织细胞的生长特性进行了比较.结果表明:在相同的体外培养条件下,胎鼠心肌细胞的整体长势良好,贴壁情况和增殖速度都较佳,有望成为胚胎干细胞饲养层的首选材料.     

DiI荧光标记示踪兔皮肤成纤维细胞修复前交叉韧带损伤的研究

体内采用组织工程方法构建前交叉韧带种子细胞筛选研究,提供了一种理想的细胞标记的方法.体外分离培养兔皮肤成纤维细胞(SF),比较皮肤成纤维细胞使用荧光染料CM-DiI体外标记前后细胞的生长状态、增殖活性以及检测SF复合纤维生物蛋白胶植入前交叉韧带上12周后的荧光表达情况.结果表明CM-Dil标记SF效果良好,标记阳性率高于95%.而标记前后SF增殖活性无明显变化,荧光标记在体内实验12周后仍有表达.从而证实CM-DiI是筛选前交叉韧带组织工程种子细胞筛选研究中细胞标记、示踪的良好标记物.     

Ⅱ型胶原诱导的关节炎大鼠滑膜成纤维细胞的培养、纯化和鉴定

类风湿关节炎(Rheumatoid arthritis,RA)病人滑膜成纤维细胞(Fibroblast-like synoviocytes,FLS)是研究RA发病机制和进行RA药物筛选的最理想的细胞来源.但是对于众多非医疗机构的高校和科研院所而言,由于RA病人滑膜组织不易获取而难以进行细胞水平的操作.因此,建立胶原诱导的关节炎大鼠(Collagen-induced arthritis,CIA)模型,成功分离膝关节滑膜组织并进行滑膜成纤维细胞的原代培养和纯化,将有效解决RA FLS的来源问题,为后续实验奠定基础.本实验通过皮内和皮下两次注射鸡Ⅱ型胶原诱导Wistar大鼠罹患关节炎,通过关节炎指数评分和膝关节HE染色明确CIA模型的建立;分离膝关节滑膜组织,采用Ⅱ型胶原酶和无EDTA胰酶消化法分离细胞并进行传代培养,传至第三代时,细胞免疫荧光检测细胞中VCAM-1蛋白含量以鉴定细胞纯度.结果显示,滑膜组织经过消化和培养后得到大量大鼠滑膜成纤维细胞;在显微镜下观察所得细胞呈梭形树突状,具有成纤维样细胞特征;VCAM-1蛋白阳性细胞达98%以上,确证所分离细胞为滑膜成纤维样细胞,且纯度极高.本研究建立了较简便快捷地获得大量滑膜成纤维细胞的方法,为RA的发病机制研究和药物防治提供了丰富的实验材料.     

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.     

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

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Achieving Unusual States of Matter Under High Pressure

The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to