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

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.     

Expression of DNA repair gene ERCC1 and its relationship with PAH-DNA adducts in lung cancer tissues

AIM: To investigate the expression of nucleotide excision repair gene ERCC1 and its relationship with PAH (polycyclic aromatic hydrocarbons)-DNA adducts in lung cancer tissues. METHODS: ERCC1 mRNA expression and the PAH-induced DNA adducts were detected in 150 lung cancer tissues, 120 adjacent lung tissues without cancer cells, 40 benign lung lesions and 40 normal lung tissues. The effects of some exposure factors on the expression of ERCC1 gene and the connection between ERCC1 and PAH-DNA adduct was analyzed. RESULTS: Reduced expression levels of ERCC1 were observed in 46 of 150 (30.7%) lung cancer specimens and 1 of 40 (2.5%) normal lung tissues. Smoking may suppress the expression of ERCC1 gene. The level of PAH-DNA adduct was negatively correlated with the expression of ERCC1 gene, the Spearman coefficient was -0.648, P<0.01. CONCLUSION: ERCC1 is an important nucleotide excision repair gene and may participate in the repair of DNA damage, such as PAH-DNA adduct. Low expression of ERCC1 may play an important role in the development of human lung cancer.     

Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta

AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.     

浅论优质牧草在肉牛饲养中的利用

针对我国肉牛饲养在饲料利用中的误区,结合肉牛的消化及代谢特点,分析了牧草的鲜喂、青贮、干草饲喂以及综合利用4种不同的利用方式及其营养特点,指出优质牧草用于肉牛饲养能充分发挥牧草的优良特性,而且符合我国畜牧业发展要求,值得广泛推广.     

川西北高寒沙地不同年限高山柳林下优势植物碳、氮、磷生态化学计量特征

通过采集川西北高寒沙地不同年限(6、18、24年)高山柳林下3种优势植被藏沙蒿、裂叶独活和镰荚棘豆,分别测定分析3种植被叶片、根部C、N、P化学计量特征变化特征。结果表明:不同年限高山柳林下植被C、N、P含量及其比值间存在显著差异且呈现出不同变化趋势。林下植被C含量整体下降;叶N含量呈上升趋势,根N含量随年限增长而下降;除藏沙蒿外,林下植被P含量变化不显著;C∶N变化范围为1.92~12.86;C∶P为29.18~196.88;不同年限高山柳林下植被N∶P间虽存在差异,但均表现出主要受到P限制,表明该区域植被生长主要受P限制,应注意P养分的适当补充。     

"三高两低"杂交油菜油研1220的选育与技术研究

油研1220是以黄籽双低隐性核不育系8078A与双低恢复系0202R配制成功的黄籽双低杂交油菜新组合。在贵州省2004~2006两年15点次试验中,增产点和增产8%以上的点分别为11个和8个,分别占总点数的73.3%和53.3%,两年平均产量达167.88 kg/667m2,比CK增产7.0%;产油量达70.72 kg/667m2,比CK增产13.87%。芥酸含量0.6%,硫甙含量28.28μmol/g.饼,含油量达44.03%,种子蛋白质含量达26.85%。选择含油率和蛋白质含量高的转育亲本,在后代分离世代中加强亲本含油率和蛋白质含量的定向选择是提高品种含油率和蛋白质含量的重要选育技术。     

植物化感作用与害虫综合治理

对植物化感作用概念的发展、植物化感物质、植物间和植物与昆虫间的化感作用进行了概述,论述了植物化感作用在害虫综合治理中的重要作用及其应用前景。     

城市草坪土壤呼吸冬季日动态特征研究

以福州市区的城市草坪为研究对象,采用Li-8100对其土壤呼吸速率进行3 d的连续观测,研究草坪土壤呼吸昼夜变化特征及其影响因子.结果表明:1)草坪土壤呼吸有明显的日变化特征;2)草坪土壤呼吸速率昼夜变化呈单峰曲线模式,雨天最大值出现在21:30,阴天最大值出现在11:30,晴天最大值出现在15:30;3)阴雨天气时土壤温度和含水量与土壤呼吸速率的双因素回归关系不显著,晴天时土壤温度与土壤呼吸速率的回归性极显著(p＜0.01),土壤温度可以解释该日土壤呼吸速率昼夜变化的70.13%,土壤含水量与土壤呼吸速率的线性关系仍不显著;土壤温度和含水量两个因素共同解释该日土壤呼吸昼夜变化的74.10%.     

热风循环智能化烤房试验

为了达到烤烟烤房操作简单,提高烤烟质量等级和烘烤自动化程度,进而提高烟农经济效益的目的,对热风循环智能化烤房的烘烤成本、初烤烟质量、效益状况等进行了试验研究。结果表明,与常规热风式烤房比较,智能化烤房在节约用煤、烘烤时间、提高烟叶质量等方面都较优;操作过程简便安全,增质节能明显,综合效益增加153.24元/667m2,经济效益明显。     

中华鳖GHITM cDNA基因克隆及表达分析

为研究GHITM基因在中华鳖(Trionyx sinensis)胚胎发育及生长中的作用,本研究通过RT-PCR和RACE方法获得了中华鳖GHITM全长cDNA序列;采用实时荧光定量PCR对GHITM mRNA组织表达及不同温度孵化胚胎发育过程中的表达特性进行分析。结果显示,中华鳖GHITM cDNA序列长度为2650 bp,开放阅读框为1050 bp,5?非编码区为123 bp,3?非编码区为1477 bp,编码349个氨基酸,编码蛋白的等电点为10.01,分子量为37.12 kDa。GHITM编码氨基酸序列由胞外区、跨膜区和胞内区组成,7个跨膜域组成跨膜区。GHITM编码氨基酸序列的同源性分析显示,中华鳖与锦龟(Chrysemys picta bellii)和绿海龟(Chelonia mydas)同属一个分支,3种鳄鱼构成一个分支,7种鸟类形成一个分支。实时荧光定量检测显示,GHITM基因在肝脏、肌肉和脑垂体中的表达水平较高,显著高于心脏、性腺、肠、肾脏和脾脏组织(P<0.05);50 g左右和500 g左右雄性个体肝脏中的GHITM基因表达量显著高于雌性个体(P<0.05);低温胁迫孵化能显著抑制胚胎肝脏中GHITM基因的表达(P<0.05)。上述研究结果表明,GHITM基因与中华鳖生长和胚胎发育密切相关,其在中华鳖胚胎中的表达受孵化温度调节。本研究为探讨中华鳖胚胎发育和生长提供理论依据。