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.     

草地生态系统碳循环研究进展

随着全球气候变化和陆地生态系统碳循环研究的发展,碳循环正成为草地生态系统研究的热点.本文在介绍其研究发展过程、主体内容和方向的同时,对目前的研究动态和研究方法进行系统的概述,重点分析了目前国内外草地生态系统碳循环过程研究、机制研究、研究方法及模型的发展动态,并对今后草地生态系统碳循环和相关研究提出了建议.     

乙烯对匍匐翦股颖ISR抗病反应中AsA-GSH循环及胼胝质沉积的影响

以匍匐翦股颖品种“Penn-A4”为试验材料,侵染立枯丝核菌后,经丁二醇(BDO)诱导产生系统抗性(ISR),喷施不同浓度的乙烯合成抑制剂氯化钴(CoCl₂)和促进剂1-氨基环丙烷羧酸(ACC)后,检测抗坏血酸-谷胱甘肽(AsA-GSH)循环中抗坏血酸(AsA)和还原型谷胱甘肽(GSH)含量及相关酶活性的变化,并观察匍匐翦股颖ISR反应中胼胝质沉积的变化。结果表明,乙烯抑制剂处理下,匍匐翦股颖幼苗中AsA含量较低,抗坏血酸过氧化物酶(APX)活性下降,大量GSH被催化还原为氧化型谷胱甘肽(GSSG),GSSG大量积累,同时谷胱甘肽还原酶(GR)活性较低,GSSG经GR少量还原为GSH。乙烯促进剂处理下,AsA含量较高,APX活性升高,GSSG在高活性GR作用下催化还原为GSH,使得GSH大量积累。因此在匍匐翦股颖ISR抗病反应中,高浓度乙烯促进AsA和GSH的大量积累,它们不仅参与了活性氧的代谢平衡,同时也作为信号分子在ISR抗病反应中起着重要作用。匍匐翦股颖感染褐斑病后,胼胝质主要沉积在厚壁细胞、韧皮部、木质部和表皮组织,其中厚壁细胞胼胝质沉积最多,表皮组织沉积最少。此外,胼胝质沉积面积在不同乙烯信号分子处理间存在差异,但随着处理时间的延长差异不显著。一定浓度的乙烯对ISR反应中胼胝质的沉积具有一定的影响,随着处理时间的延长显著增加,在100 μmol·L^-1ACC处理5 d后,胼胝质沉积总面积仅为9.916 mm²,处理10 d后升至最高,为38.396 mm²,但在病害侵染后期,胼胝质数量减少,在100 μmol·L^-1ACC处理15 d后,胼胝质沉积总面积降至20.052 mm²,反映了乙烯信号分子对胼胝质沉积的影响是一种短期效应,短期内可提高匍匐翦股颖植株抗病性,具有信号分子的短时效特点。研究结果为探清匍匐翦股颖ISR抗病响应中ET信号分子如何调控抗病生理特性提供了理论基础。     

The localization and expression of epidermal growth factor and epidermal growth factor receptor in bovine ovary during oestrous cycle

Epidermal growth factor (EGF) is one of the important regulatory factors of EGF family. EGF has been indicated to effectively inhibit the apoptosis of follicular cells, to promote the proliferation of granulosa cells and the maturation of oocytes, and to induce ovulation process via binding to epidermal growth factor receptor (EGFR). However, little is known about the distribution and expression of EGF and EGFR in cattle ovary especially during oestrous cycle. In this study, the localization and expression rule of EGF and EGFR in cattle ovaries of follicular phase and luteal phase at different time points in oestrous cycle were investigated by using IHC and real-time qPCR. The results showed that EGF and EGFR in cattle ovary were mainly expressed in granulosa cells, cumulus cells, oocytes, zona pellucida, follicular fluid and theca folliculi externa of follicles. The protein and mRNA expression of EGF/EGFR in follicles changed regularly with the follicular growth wave both in follicular and in luteal phase ovaries. In follicular phase ovaries, the protein expression of EGF and EGFR was higher in antral follicles than that of those in other follicles during follicular growth stage, and the mRNA expression of EGFR was also increased in stage of dominant follicle selection. However, in luteal phase ovaries, the growth of follicles was impeded during corpus luteum development under the action of progesterone secreted by granular lutein cell. The mRNA and protein expressions of EGF and EGFR in ovarian follicles during oestrous cycle indicate that they play a role in promoting follicular development in follicular growth waves and mediating the selection process of dominant follicles.     

Environmental performances of giant reed (Arundo donax L.) cultivated in fertile and marginal lands: A case study in the Mediterranean

Perennial rhizomatous grasses (PRGs) tend to have a high yield combined with a low environmental impact. Cultivation in marginal or poorly cultivated land is recommended in order not to compromise food security and to overcome land use controversies. However, the environmental impacts of using different types of soil are still unclear. We thus assessed the environmental impact of two giant reed (GR) systems cultivated in a fertile soil (FS) and in a marginal soil (MS) through a cradle-to-plant gate LCA. We analyzed energy balance, GHG emissions (including LUC, not including iLUC), and the main impacts on air, water and soil quality. In both systems the annualized soil carbon sequestration was more than twofold the total GHG emitted, equal to −6464 kg CO₂eq ha⁻¹ in FS and −5757 kg CO₂eq ha⁻¹ in MS. Overall, soil characteristics affected not only GR yield level, but also its environmental impact, which seems to be higher in the MS system both on a hectare and tonne basis. The production of GR biomass in marginal soil could thus lead to higher environmental impacts and a more extensive land requirement.     

铲头堆砂白蚁原始繁殖蚁形成的周期

铲头堆砂白蚁Cryptotermes declivis Tsai et Chen是破坏木材及木制品的重要干木白蚁,其有翅成虫通过分飞配对后建立新群体,是其重要繁殖扩散途径。本文对铲头堆砂白蚁原始繁殖蚁形成周期进行了研究。以似桂木Paratocarpus sp.、团花Anthocephalus chinensis(Lam.) Rich. et Walp.和球花豆Parkia sp.作为该种白蚁食料,研究其原始繁殖蚁配对后从新建群体的形成到产生下一代的原始繁殖蚁,完成1个世代生活周期。结果表明,铲头堆砂白蚁在室温下,新建群体的形成、发展、发育至下一代原始繁殖蚁形成的周期约5 a,不同寄主材料对铲头堆砂白蚁生活周期没有显著性影响,但与群体数量多少显著相关。本研究首次明确了铲头堆砂白蚁群体发育成熟的周期,对该害虫的检疫与防控研究具有重要参考价值。     

Energy use and greenhouse gas emissions from turf management of two Swedish golf courses

Turf management on golf courses entails frequent maintenance activities, such as mowing, irrigation and fertilisation, and relies on purchased inputs for optimal performance and aesthetic quality. Using life cycle assessment (LCA) methodology, this study evaluated energy use and greenhouse gas (GHG) emissions from management of two Swedish golf courses, divided into green, tee, fairway and rough, and identified options for improved management. Energy use and GHG emissions per unit area were highest for greens, followed by tees, fairways and roughs. However, when considering the entire golf course, both energy use and GHG emissions were mainly related to fairway and rough maintenance due to their larger area. Emissions of GHG for the two golf courses were 1.0 and 1.6 Mg CO₂e ha⁻¹ year⁻¹ as an area-weighted average, while the energy use was 14 and 19 GJ ha⁻¹ year⁻¹. Mowing was the most energy-consuming activity, contributing 21 and 27% of the primary energy use for the two golf courses. In addition, irrigation and manufacturing of mineral fertiliser and machinery resulted in considerable energy use. Mowing and emissions associated with fertilisation (manufacturing of N fertiliser and soil emissions of N₂O occurring after application) contributed most to GHG emissions. Including the estimated mean annual soil C sequestration rate for fairway and rough in the assessment considerably reduced the carbon footprint for fairway and turned the rough into a sink for GHG. Emissions of N₂O from decomposition of grass clippings may be a potential hotspot for GHG emissions, but the high spatial and temporal variability of values reported in the literature makes it difficult to estimate these emissions for specific management regimes. Lowering the application rate of N mineral fertiliser, particularly on fairways, should be a high priority for golf courses trying to reduce their carbon footprint. However, measures must be adapted to the prevailing conditions at the specific golf course and the requirements set by golfers.     

补光对兰云杉和红皮云杉生长的影响

云杉苗期生长缓慢是生产中的主要问题,补光可促进苗木生长,缩短育苗周期,提高育苗效率.研究不同光源(阳光灯、镝灯、碘钨灯)、不同光照时间对兰云杉和红皮云杉1年生苗苗高、地径、侧枝数、根系、生物量等生长性状的影响,结果表明:光照处理可以抑制苗木休眠,促进其持续生长,对2种云杉的各生长指标均有显著的促进作用；不同光源、不同光...     

战后美国经济周期再考察

二战后美国经济周期波动发生显著变化。相比二战之前，美国经济周期扩张期拉长，衰退期缩短，二者之间形成剪刀差。经济周期长度、扩张期跨度和紧缩期跨度的离散度远远大于战前。二战后至今，美国经济中周期出现（朱格拉周期）呈拉长的趋势，扩张期远远长于衰退期，中周期（朱格拉周期）和短周期（基钦周期）的波幅经历了从小到大的阶段性变化。     

日粮鱼油对高脂日粮饲喂小鼠发情周期和机体产热的影响

本研究旨在探讨鱼油对高脂日粮饲喂小鼠发情周期和机体代谢产热的影响。试验选用36只4周龄C57BL/6 J雌性小鼠,随机分成3组（n=12）：对照组、高脂组和高脂+鱼油组。对照组饲喂标准啮齿动物饲料（AIN-93G）,高脂组和高脂+鱼油组分别饲喂高脂日粮（脂肪提供60%能量）和添加5%鱼油（等能替代猪油）的高脂日粮。试验期间,对小鼠体组成（12周龄）、整体代谢（16周龄）、褐色脂肪温度（18周龄）、体核温度（直肠温度,18周龄）和发情周期（20周龄）等进行检测。试验结束后,眼球采血分离血清,检测促卵泡激素（follicle-stimulating hormone,FSH）和雌二醇（estradiol,E2）的水平。此外,采集皮下脂肪、腹部脂肪和肩胛间褐色脂肪,称重并使用Western blot检测脂肪组织中产热相关基因的蛋白表达（UCP1、Cyto C）,使用实时荧光定量PCR检测褐色脂肪组织中产热基因的mRNA表达（UCP1, PRDM16,PGC1α,Cidea,Elovl3）。结果显示,与对照组相比,高脂日粮显著增加了小鼠的体脂含量（12周龄）及皮下和腹部脂肪的沉积量（21周龄）（P<0.05）,而添加鱼油显著降低了高脂饮食引起的体脂含量增加（P<0.05）。另外,高脂日粮导致小鼠的发情周期紊乱,伴随着周期延长、发情期缩短,以及血清中FSH和E2的水平降低（P<0.05）,而添加鱼油可缓解高脂日粮导致的小鼠发情周期紊乱,提高血清中FSH和E2的水平（P<0.05）。同时,添加鱼油可增加高脂饲喂小鼠肩胛间褐色脂肪（interscapular brown adipose tissue,iBAT）和腹股沟白色脂肪（inguinal white adipose tissue,iWAT）中产热相关基因的表达（P<0.05）,进而促进iBAT激活/产热和iWAT褐色化。结果提示,日粮鱼油可缓解高脂日粮导致的发情周期紊乱,可能与BAT激活和WAT褐色化造成的机体代谢产热增强有关。