Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes     

JIANG Xun  ZENG Yao-ying  HE Xian-hui  XU Li-hui  DI Jing-fang  FENG Zheng  ZHAO Jing-xian  WANG Qing  WANG Tong  SHI Jian-bo 《园艺学报》2004,20(6):924-928

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.  相似文献   

Meiotic competence of mouse oocytes reconstructed by replacing the germinal vesicle with a male pronucleus and somatic nucleus     

Honglin LIU  Fugaku AOKI 《Animal Science Journal》2004,75(2):125-129

The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes.  相似文献   

Significance of local immunity in hen reproductive organs     

Yukinori YOSHIMURA 《Animal Science Journal》2004,75(3):183-191

The current paper describes aspects of local immunity in the ovary and oviduct, and the significance of immunity to reproductive functions in hens. The immunocompetent cell populations in the ovary and oviduct change with a positive correlation to sexual activity, and gonadal steroid is one of the key factors in the increase. Local immune responses mediated by major histocompatibility complex class II and T cell subsets occur in response to infection by Salmonella enteritidis, which may contaminate eggs. In the ovary, immunocompetent cells are also suggested to play roles in the regulation of ovarian functions. Macrophages and T cells are likely to enhance the regression of atretic follicles to maintain the ovarian tissue microenvironment. Autoantibodies to ovarian tissues appeared in the hens with low egg laying frequency, suggesting that the auto‐antibodies may be one of the factors in the decline of egg production. In the oviduct, local immunity possibly has a role in the selection of sperm, though the immunoreactions may also affect sperm survival leading to the decline in fertility. The concentration of yolk IgY, which plays a role in maternal immunity transmission, significantly decreases with the aging of birds, whereas it is significantly increased by estrogen. Therefore, the immune system plays significant roles not only in defense against infection, but also in the functions of reproductive organs. Investigations on the local immune system in the reproductive organs and factors affecting it are of importance for the production of sterile eggs and improvement of reproductive functions.  相似文献   

贺兰山西坡不同海拔梯度草地土壤氮特征   总被引：9，自引：1，他引：9  

傅华  裴世芳  张洪荣 《草业学报》2005,14(6):50-56

以阿拉善左旗境内贺兰山中段(西坡)及山前地带不同海拔下的草地为研究对象,分析不同海拔梯度下草地土壤全氮的分布特征及其与气候因子、植被特征和土壤特性的关系.结果表明,土壤全氮含量随海拔的降低而逐渐降低,在典型草原中沿土壤垂直剖面依次降低,在高寒草甸、荒漠化草原和草原化荒漠0～20与20～40 cm土层中差异不显著,但高于40～60 cm土层.在海拔1 360～1 380 m草原化荒漠的5个样地上,由于放牧利用程度和植被类型的不同,0～20 cm土层土壤氮的含量变幅在0.022 8%～0.034 7%.0～20,20～40和40～60 cm土层土壤全氮含量与年降水量、植被盖度、草地生产力、土壤含水量、有机碳含量和<0.05 mm颗粒含量呈极显著正相关,与年均温、土壤pH值呈极显著负相关(P<0.001).偏相关分析显示,影响0～20 cm土层全氮(TN)含量最主要的因素是植被盖度(C)和年降水量(AP),相关方程是TN%=0.002 040 C 0.000 585 AP-0.076 800;而影响20～40 cm土层全氮含量的最主要因素是草地生产力(B),与土壤全氮回归方程为TN%=0.001 6 B 0.024 7.而各环境因素中影响土壤全氮含量最主要的因素,0～20 cm土层为土壤有机碳和年降水量,20～40 cm土层主要受土壤有机碳的影响.  相似文献   

光氮互作对草地早熟禾碳氮代谢的影响     

姜丹  陈雅君  刘丹  胡海辉 《中国草地学报》2005,27(6):49-53

以草地早熟禾的3个品种为试验材料,研究了高低光照下不同的施氮水平对植株体内碳氮代谢的影响。对可溶性糖和硝酸还原酶的含量测定结果显示:高低光强下早熟禾的氮素同化能力均随着氮水平的升高呈现先升高后降低的趋势,碳代谢的中间产物可溶性糖随着氮水平的升高呈现先降低后升高的趋势,高低光下糖的含量随着氮水平的升高其品种间差异逐渐减小;在低氮条件下,植物的氮素同化能力较弱,光合碳代谢相对较强,糖含量较高,随着氮水平的增大,光合产物逐渐流向氮素代谢,硝酸还原酶含量升高。  相似文献   

光照和褪黑激素对内蒙古绒山羊氮分配的影响   总被引：1，自引：0，他引：1  

王林枫  卢德勋  孙海洲  赵秀英  珊丹 《畜牧兽医学报》2005,36(12):1300-1306

从氮分配的角度研究了光照和褪黑激素对内蒙古白绒山羊营养分配的影响。结果表明:光照时间和埋植褪黑激素显著影响绒山羊体内氮物质分配,短光照或埋植褪黑激素显著提高绒山羊血液中的褪黑激素水平,并使其他相关激素如催乳素(PRL)、胰岛素(INS)、类胰岛素生长因子-I(IGF-I)、瘦素(LEP)的含量发生显著变化,结果使毛绒氮的沉积增加,体氮的沉积减少。短光照条件下毛绒氮和体氮的沉积分别为33.7%±0.64%和66.3%±0.64%;而长光照条件下则减少毛绒氮的分配量,增加体氮分配量,毛绒氮和体氮的沉积分别为23.6%±0.46%和76.4%±0.46%。短光照和褪黑激素之间有强烈的互作效应,短光照埋植褪黑激素组毛绒氮和体氮的分配比例分别为36.1%±0.79%和63.9%±0.79%。试验期绒山羊的产绒量平均增加(338.83±72)g,比普通绒山羊提高73.86%,新生羊绒的品质符合纺织工业标准的要求。  相似文献   

母牛一胎双犊技术的研究     

张居农  蒋学国  许斌  郭成宽  仇其山  李强  牛志宏  王安江  王凤娇  张建军  张选珠 《乳业科学与技术》2005,27(3):137-138

选择9头母牛先行人工授精、7天后再移植冷冻胚胎以生产双犊。试验结果表明,母牛产双犊达44.44%(P<0.01)。提示:采用A(I人工授精)+ET(胚胎移植)是提高母牛双犊率的有效技术措施之一。  相似文献   

Likelihood of pregnancy after embryo transfer is reduced in recipient mares with a short preceding oestrus          下载免费PDF全文

J. Cuervo‐Arango  A. N. Claes  M. Ruijter‐Villani  T. A. Stout 《Equine veterinary journal》2018,50(3):386-390

  相似文献   

不同施肥模式对玉米各器官碳氮累积和分配的影响   总被引：3，自引：0，他引：3  

谢军  徐春丽  陈轩敬  王珂  李丹萍  张跃强  石孝均 《草业学报》2018,27(8):50-58

以长期定位试验为依托,选取:1)不施肥(CK);2)农民习惯施肥(FP);3)推荐施肥(OP);4)有机肥氮替代100%化肥氮(OM);5)有机肥氮替代50%化肥氮(MF)5个处理,研究不同施肥模式对玉米植株及各器官碳氮含量、碳氮分配比例及C/N的影响,为西南紫色土地区合理施肥、作物增产提供科学依据。结果表明,与FP处理相比,MF处理显著增加了玉米植株生物量,达26.2%。相比OM和OP处理,MF处理显著增加玉米苞叶和根茬中碳浓度,分别增加5.4、4.2 g·kg^-1和7.4、21.3 g·kg^-1,同时增加玉米苞叶、根茬、穗轴和籽粒中的碳储量,玉米茎秆和籽粒中的氮储量也有增加。此外,相比FP处理,MF处理能显著增加玉米整株的碳储量和氮储量,达29.1%和16.9%。等氮水平下,MF、OP处理均能增加玉米苞叶和籽粒中碳同化物的分配比例,MF处理玉米籽粒中氮素的分配比例较OP和OM处理分别增加1.7%和3.6%,同时MF处理能使玉米维持较高的C/N。综上,有机肥氮替代50%化肥氮能增加玉米植株的生物量,同时提高玉米对氮素的吸收和碳素的累积,增加玉米籽粒中碳同化物和氮素的分配比例,同时,有机肥氮替代50%化肥氮能使玉米植株维持较高的C/N,有利于产量的形成,该施肥方式不仅能够促进氮素的高效利用,减少化肥的投入,还能够减少化肥损失,降低氮素损失引发的环境风险。  相似文献   

水氮供应对地下滴灌紫花苜蓿生长特征及草地小气候的影响     

胡伟  张亚红  李鹏  刘瑞  蔡伟  王小菊 《草业学报》2018,27(12):122-132

为探讨不同水氮处理条件下紫花苜蓿生长状况与草地小气候特征的关系,以2年生紫花苜蓿“巨能7号”为研究对象,采用田间试验和室内分析相结合的方法,研究了宁夏引黄灌区地下滴灌条件下不同滴灌量和施氮量处理下紫花苜蓿生长特征和草地小气候的变化。结果表明:1)滴灌量和施氮量对紫花苜蓿的株高、叶面积和鲜草产量都有显著的影响,表现为紫花苜蓿的株高、叶面积和鲜草产量均随滴灌量和施氮量的增加而增加,当施氮量增加到一定值时,继续增施氮肥,其鲜草产量增产效果在不同滴灌量处理下表现出不同的趋势。2)与不施氮处理相比,增施氮肥降低了紫花苜蓿株间空气温度、浅层土层温度和株间光照强度,而增加了群体内部空气相对湿度。3)不同滴灌量对紫花苜蓿的生长微环境的调节作用不同,随着滴灌量的增加,紫花苜蓿群体相对湿度逐渐提高,而紫花苜蓿株间气温和浅层土层温度降温效应越明显。4)紫花苜蓿生育期间株高与叶面积、草产量和群体内部相对湿度呈极显著正相关(P<0.01),与群体内光照强度、株间气温、浅层土壤温度呈极显著负相关(P<0.01)。合理减少滴灌量和施氮量不仅能维持紫花苜蓿良好的生长特征,而且能提高鲜草产量和改善草地生态环境条件。本研究旨在为紫花苜蓿群体微环境生态因子的改善及高产优质栽培措施提供科学依据。  相似文献