栉孔扇贝血细胞的显微和超微结构

通过光镜、扫描电镜和透射电镜观察栉孔扇贝Chlamys farreri血细胞的形态、结构特征,对其分类进行了初步研究。根据细胞的大小、形态和结构特征可将血细胞分成5种类型:Ⅰ型小透明细胞,大小约2.44±0.11μm,约占45%—50%;Ⅱ型大透明细胞,大小约4.83±0.28μm,约占15%—20%;Ⅲ型小颗粒细胞,大小约4.07±0.15μm,约占15%—20%;Ⅳ型中颗粒细胞,大小约7.20±0.26μm,约占20%—25%和Ⅴ型大颗粒细胞,大小约13.87±0.73μm,约占3%—5%。血细胞在血淋巴中的平均密度为(3.03±0.11)×107cell.ml—1,其中颗粒细胞占42.6%,透明细胞占57.4%。透射电镜观察结果表明,颗粒细胞的细胞质颗粒可区分成3种类型:Ⅰ型高电子密度颗粒,Ⅱ型低电子密度颗粒和Ⅲ型中等电子密度颗粒。

神经垂体激素颗粒释放形式和转运途径的结构基础(英文)

背景:神经垂体多肽激素分泌颗粒的释放形式和在细胞外正常转运途径的结构基础是以胞吐分泌方式还是浓度差因素影响而进入脑脊液的。目的:通过观察大鼠神经垂体的结构特征,为神经垂体多肽激素分泌颗粒的释放形式和在细胞外正常转运途径的结构基础提供形态学依据。设计:随机对照实验。单位:一所大学的人体解剖学教研室。材料:实验于2003-05/2004-01在河北医科大学人体解剖学教研室完成。选择健康成年雄性SD大鼠12只,体质量约300g,清洁级,由河北省实验动物中心提供。方法:将大鼠随机分成3组,每组4只,分别进行神经垂体光镜、透射和扫描电镜观察。主要观察指标:神经垂体的显微和超微结构。结果:12只大鼠全部进入结果分析。在光镜下观察,大鼠垂体冠状切面上,可显示垂体3部分的结构关系,远侧部,中间部及神经部清晰可辨。在透射电镜下观察,神经垂体主要由无髓神经纤维、垂体细胞及富含毛细血管的结缔组织组成。毛细血管内皮为有孔型(50nm),外有基膜与血管周隙分隔。完整的大型膜包分泌颗粒(100～300nm)不仅大量存在于无髓神经纤维内,而且也见于血管周隙内。在扫描电镜下观察,垂体囊由单层扁平上皮细胞和上皮下结缔组织构成,囊上皮细胞间存在许多不规则的囊上皮孔(2～5μm),上皮孔附近经常可见分泌颗粒。结论:神?

IPP5对高糖高胰岛素诱导的心肌细胞肥大的影响

目的:探讨体外培养中I型磷酸酶的新型抑制亚基(IPP5)对高糖高胰岛素诱导的心肌细胞的肥大是否具有抑制作用。方法:新生SD乳鼠心肌细胞在含有10%胎牛血清的杜尔伯科改良伊格尔培养基(DMEM)中培养72h后,分别将IPP5野生型和短片段活化突变型基因通过脂质体介导转染入心肌细胞,48h后,加入高糖高胰岛素作用48h。利用图像分析软件分析心肌细胞表面积,并检测心肌细胞总蛋白含量、肌酸激酶(CK)和乳酸脱氢酶(LH)漏出量及心肌损伤标志物心肌肌钙蛋白I(TnI)的变化。结果:高糖高胰岛素可增加心肌细胞表面积,IPP5突变活化型基因可部分抑制心肌细胞肥大;转染IPP5突变活化型基因组的心肌细胞总蛋白含量较转染空载体对照组高,而CK和LH漏出量及TnI的含量却比转染空载体对照组明显减少。此外,转染IPP5突变活化型基因组的心肌细胞形态轮廓清楚,细胞质颗粒少,而转染空载体对照组的心肌细胞轮廓不清,细胞质颗粒粗大且多。结论:IPP5突变活化型对高糖高胰岛素诱导的心肌细胞肥大、损伤具有一定的抑制保护作用。

虹鳟鱼血细胞的观察

虹鳟(Salmo gairdneri irideus)是鲑科鱼类中适应性很广的冷水性鱼种,常年生活在低温的流水中,它原产于美国,后来通过人工驯化,现已遍布世界各地。我国于1959年由朝鲜引进,相继在十多个省市养殖成功。由于虹鳟鱼的营养丰富、肉质细嫩、味道鲜美,而成为一种供不应求的高档鱼种,具有很高的经济价值。但在养殖过程中若是饲养及管理条件不良。

氦-氖激光对人大颗粒淋巴细胞内嗜天青颗粒影响的图像分析研究

目的探讨氦－氖（ＨｅＮｅ）激光对人体的免疫刺激作用的机理。方法用３．１Ｊ／ｃｍ２、９．４Ｊ／ｃｍ２、２５．０Ｊ／ｃｍ２和４９．９Ｊ／ｃｍ２剂量的ＨｅＮｅ激光分别在体外辐照人外周血，细胞化学方法显示大颗粒淋巴细胞内嗜天青颗粒，并用图像分析系统对嗜天青颗粒作定量分析。结果３．１Ｊ／ｃｍ２辐照组的嗜天青颗粒数目、颗粒面积、颗粒面积与细胞面积比值和颗粒积分光密度四项参数值与对照组比较无明显变化；９．４Ｊ／ｃｍ２辐照组的四项参数值明显增高；２５．０Ｊ／ｃｍ２辐照组的四项参数值进一步增高；４９．９Ｊ／ｃｍ２辐照组的四项参数值较２５．０Ｊ／ｃｍ２辐照组明显降低。结论适当剂量的ＨｅＮｅ激光能活化人大颗粒淋巴细胞内嗜天青颗粒，提高机体的免疫功能；最适激光剂量的选择是临床应用中应考虑的一个重要因素。

Improvement of in vitro Cytoplasmic Maturation of Denuded Porcine Oocytes by Coculture with Follicular Mural Granulosa Cells

[Objective] This study aimed to improve the in vitro maturation quality of denuded porcine oocytes and provide scientific basis for establishing a stable and efficient denuded oocyte culture system. [Method] The first polar body extrusion rate, oocyte glutathione （GSH） content, positive rate of brilliant cresyl blue （BCB） staining and development potential of activated oocytes or fertilized oocytes were employed as main indicators to investigate the effects of follicular mural granulosa cell （MGC） coculture on cytoplasmic maturation of cumulus cell-removal oocytes （Denuded Oocyte, DO）. [Result] According to in vitro maturation results, compared with DO group, the first polar body extrusion rate of porcine oocytes in DO＋MGC group was not significantly different, but the nuclear maturation process was improved and was more similar to that in COC （cumulus-oocyte complex） group. Detection of GSH content in mature oocytes showed that there was no significant difference between DO＋ MGC group （optical density of 1 053.67） and COC group （optical density of 1 426.00） or between DO＋MGC group and COC＋GC group （optical density of 1 541.00）, however, GSH content in mature oocytes of DO group （optical density of 724.67） was significantly lower than that of COC group and COC＋GC group （P0.05）. Detection of glucose-6-phosphate dehydrogenase （G6PDH） activity showed that there was no significant difference in BCB positive oocyte rate between DO ＋MGC group （88.26% ） and COC group （92.75%） or between DO＋MGC group and DO group （82.86% ）, however, BCB positive oocyte rate of DO group was significantly lower than that of COC group （P0.05）. Furthermore, the cleavage rate and blastocyst rate of activated mature oocytes derived from DO ＋MGC group （94.98% and 43.67% , respectively） were significantly higher than those from DO group （52.54% and 8.97%, respectively） （P0.05）, and were not significantly different compared with those from COC group （97.11% and 38.30%, respectively）. In addition, the cleavage rate of fertilized oocytes derived from DO＋MGC group （72.65%） showed no significant difference compared with that from DO group （63.59%）, but the blastocyst rate of DO＋MGC group was significantly higher than that of DO group （9.88%） （P0.05）. [Conclusion] MGC coculture can significantly improve the in vitro cytoplasmic maturation quality of denuded porcine oocytes, thereby enhancing the subsequent developmental potential.

The structure of the nucleosome core particle of chromatin in chicken erythrocytes visualized by using atomicforce microscopy

The structure of the nuclosome core particle of chromatin in chicken erythrocytes has been examined by usingAFM. The 146 hp of DNA wrapped twice around the corehistone octamer are clearly visualized. Both the ends ofentry/exit of linker DNA are also demonstrated. The dimension of the nucleosome core particles is- 1-4 um inheight and ～13-22 um in width. In addition, superbeads(width of - 48-57 urn, beight of-2-3 nm) are occasionallyrevealed, two turns of DNA around the core particles arealso detected.

弓形虫GRA6可溶性蛋白在大肠埃希菌中的表达及纯化

目的在大肠埃希菌中高效表达及纯化弓形虫GRA6抗原,为制作弓形虫感染基因工程诊断试剂盒奠定基础。方法将重组pGEX-GRA6表达载体转化大肠埃希菌BL21-Codon Plus (DE3)-RP菌株,在异丙基硫代-β-D半乳糖苷(IPTG)诱导下表达。超声破壁后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表达产物的表达形式,并对表达产物以硫酸铵沉淀、Sephedax G50脱盐和GST亲和层析柱进行目的蛋白的纯化。通过Western blotting检测其纯化的重组抗原的免疫反应性。结果GRA6以融合蛋白(GST-GRA6)的形式在大肠埃希菌中得到了高效表达。对表达产物可溶性分析表明,表达蛋白在上清液中和包涵体中均有表达。在上清液中表达的可溶性蛋白经纯化后蛋白纯度可达90%以上。Western blotting分析表明纯化蛋白能被弓形虫感染的人血清所识别。结论GRA6在大肠埃希菌中得到了高效表达,重组抗原经纯化后能特异性地被弓形虫感染者血清所识别。

Uptake of magnetic nanoparticles for adipose-derived stem cells with multiple passage numbers

With the increasingly promising role of nanomaterials in tissue engineering and regenerative medicine, the interaction between stem cells and nanoparticles has become a critical focus. The entry of nanoparticles into cells has become a primary issue for effectively regulating the subsequent safety and performance of nanomaterials in vivo. Although the influence of nanomaterials on endocytosis has been extensively studied, reports on the influence of stem cells are rare.Moreover, the effect of nanomaterials on stem cells is also dependent upon the action mode. Unfortunately, the interaction between stem cells and assembled nanoparticles is often neglected. In this paper, we explore for the first time the uptake of γ-Fe2O3 nanoparticles by adipose-derived stem cells with different passage numbers. The results demonstrate that cellular viability decreases and cell senescence level increases with the extension of the passage number. We found the surface appearance of cellular membranes to become increasingly rough and uneven with increasing passage numbers. The iron content in the dissociative nanoparticles was also significantly reduced with increases in the passage number. However, we observed multiple-passaged stem cells cultured on assembled nanoparticles to have similarly low iron content levels. The mechanism may lie in the magnetic effect of γ-Fe2O3 nanoparticles resulting from the field-directed assembly. The results of this work will facilitate the understanding and translation of nanomaterials in the clinical application of stem cells.