不同贮藏年限老芒麦种胚线粒体抗氧化功能的变化规律

种子在自然贮藏过程中常常伴随着内部生理机能的恶化，线粒体作为种子内活性氧（reactive oxide species，ROS）产生的主要位点是最先遭到破坏的细胞器。为探讨不同贮藏年限对老芒麦种胚线粒体抗氧化功能的影响，本试验以室温贮藏0～4年的老芒麦种子为材料，分析比较其老化规律及种胚线粒体抗氧化特性的变化规律。结果表明：随着贮藏年限的延长，老芒麦种子发芽势、发芽率和种苗鲜重逐渐下降，死种子逐渐增多，种胚线粒体苹果酸脱氢酶（mitochondria malate dehydrogenase，MDH）、谷胱甘肽还原酶（glutathione reductase，GR）、单脱氢抗坏血酸还原酶（monodehydroascorbate reductase，MDHAR）活性逐渐下降，但在死种子中超氧化物歧化酶（superoxide dismutase，SOD）活性显著升高。此外，在贮藏过程中老芒麦种胚线粒体O₂^·-产生速率不断上升，而H₂O₂含量则呈现逐渐降低的趋势，表明线粒体中O₂^·-的积累与细胞氧化损伤密切相关。     

猪胚胎干细胞的分离克隆

本文从分化抑制物、培养基的选择内细胞团的分离,胚胎干细胞的分离及鉴定方法等方面系统地综述了猪胚胎干细胞分离克隆的研究进展;并对其未来的应用前景进行展望。     

长寿花胚性愈伤组织的诱导及胚状体再生

 研究了长寿花胚性愈伤组织的筛选，胚状体的诱导发生、发育过程及植株再生。经过对外观及细胞学观察，看出外观质地疏松、颗粒状的淡黄色愈伤组织细胞圆形且形状规则，是胚性愈伤组织。诱导胚性愈伤组织的最适培养基为：MS+2，4-D 2 mg·L^-1 +BA 0.2 mg·L^-1；胚状体的诱导培养基为MS+BA 2 mg·L^-1 +NAA 0.2 mg·L^-1 +活性炭4 g·L^-1+蔗糖30 g·L^-1，胚状体诱导率可达185个胚状体；胚状体的再生培养基为MS+蔗糖30 g·L^-1。利用石蜡切片对胚状体的发生过程进行了观察。     

Comparison of different conditions inducing embryonic stem cells

AIM: To evaluate the different conditions inducing mouse embryonic stem cells (ESC) in vitro to differentiate into cardiomyocytes. METHODS: BRL conditioned medium was used to promote the growth of ESC and maintain them in an undifferentiated state. During the inducing process, retinoic acid (RA), DMSO, activin-A and TGF-β₁ were used as inducing reagents, and made up six kinds of differentiating medium. Then a three-step method inducing ESC cultured in hanging drops, in suspension and in plating was used to induce the differentiation of ESC. RESULTS: ESC were induced in vitro to differentiate into cardiomyocytes. Of all groups, the highest differentiating rate was observed in the group induced by activin-A (20 μg/L) and TGF-β₁ (2 μg/L). CONCLUSION: The inducing conditions including activin-A (20 μg/L) and TGF-β₁ (2 μg/L) is very valuable in inducing ESC differentiation into cardiomyocytes.     

Construction of mouse ESC line stably expressing Pdx1 by Tet-On system

AIM To construct the mouse embryonic stem cell （ESC） line with stable pancreatic and duodenal homeobox 1 （Pdx1） expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells. METHODS The Pdx1-overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups： blank control group （ESC group）, empty lentivirus control group （PDX1^- ESC group） and Pdx1 lentivirus transfection group （PDX1⁺ ESC group）. Flow cytometry was used to detect the transfected cells after screening by doxycycline （DOX）. The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS （1） The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. （2） With DOX, green fluorescence was observed in PDX1^- ESC group and PDX1⁺ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1⁺ ESC group （P<0.05）. Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed （P>0.05）. （3） After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells.     

Analysis of chromosome aberration due to ethidium bromide using AFM

AIM: To study chromosome aberration due to ethidium bromide (EB),a heterocyclic organic compound and an organic fluorescence dye commonly used in biochemical experiment,and to help further understanding the molecular mechanism of tumor or cancer induced by EB and other heterocyclic organic compounds.METHODS: The toxicity action of EB was evaluated from three aspects including DNA,chromosome and embryo stem cells (ESCs) using atomic force microscopy (AFM),and thereinto,the morphology structural difference of ESCs treated with two EB doses was also valuated.RESULTS: The morphological structures of DNA,chromosome and ESCs were dramatically damaged.The average height of DNA decreased 0.5 nm;chromosomal arms were ruptured from centromere location;molecules of cellular membrane congregated and loop-like structure formed,and ES cell masses were collapsed and became dead after large EB doses treatment and mesh-like morphological structure was discernable.CONCLUSION: The toxicity action of EB is strong and destroys the surface structure of DNA and chromosome.EB induces structural aberration of ES cellular membrane and cell death.The results indicate that the action of EB is externalized at gene level and cell level,which is important to study the carcinogenicity of EB.     

家蚕胚胎伴性温敏性的遗传研究

试验用新九与伴1等4个温敏性的品种杂交，将其F1代蚕种用高温干燥条件催青，结果雄蚕正常孵化，雌蚕几乎不孵化，表明伴1等品种具有控制催青温敏性表达的基因存在。用华1与伴1组配的F1、F2及F1与亲本回交的14个组合作遗传分析，认为催青敏感性状由位于Z性染色体上的一个主基因控制，呈隐性的伴性遗传。     

Na?ve-like conversion enhances the difference in innate in vitro differentiation capacity between rabbit ES cells and iPS cells

Quality evaluation of pluripotent stem cells using appropriate animal models needs to be improved for human regenerative medicine. Previously, we demonstrated that although the in vitro neural differentiating capacity of rabbit induced pluripotent stem cells (iPSCs) can be mitigated by improving their baseline level of pluripotency, i.e., by converting them into the so-called “naïve-like” state, the effect after such conversion of rabbit embryonic stem cells (ESCs) remains to be elucidated. Here we found that naïve-like conversion enhanced the differences in innate in vitro differentiation capacity between ESCs and iPSCs. Naïve-like rabbit ESCs exhibited several features indicating pluripotency, including the capacity for teratoma formation. They differentiated into mature oligodendrocytes much more effectively (3.3–7.2 times) than naïve-like iPSCs. This suggests an inherent variation in differentiation potential in vitro among PSC lines. When naïve-like ESCs were injected into preimplantation rabbit embryos, although they contributed efficiently to forming the inner cell mass of blastocysts, no chimeric pups were obtained. Thus, in vitro neural differentiation following naïve-like conversion is a promising option for determining the quality of PSCs without the need to demonstrate chimeric contribution. These results provide an opportunity to evaluate which pluripotent stem cells or treatments are best suited for therapeutic use.     

Status of Biological Studies and Aquaculture Development of the Mud Crab, <Emphasis Type="Italic">Scylla serrata</Emphasis>, in China: An Experimental Ecological Studies

This is an initial paper in a series of overviews of biological research and aquaculture development of the mud crab, Scylla serrata, in China. Results of experimental ecological studies on mud crabs are reported here. As a result of these experimental studies, results that are important for mud crab culture were also discovered and these include, methods to condition and manage broodstock, determination of ecological conditions that are suitable for embryonic development, and the influence of temperature, salinity, diet and larval density on development and survival of larvae. Results of this work will be useful in establishing a good method for artificial mass culture of larvae.     

鲶繁殖生物学的研究

嘉陵江鲶的卵母细胞划分为６个时相。第Ⅱ时相卵母细腻外不仅具有质膜，而且还有滤泡膜的结缔组织膜，第Ⅳ时相卵终细腻具漏斗状和精孔及椭圆形的精孔细腻。嘉陵江鲶的成熟年龄大多为１龄。生殖期雌雄经为２：１。繁殖季节中雌鱼的成熟系数高达１４．５％。对鲶进行人工催产，成功地获得了鲶的受精卵。鲶的受精卵圆形、绿色，膨大后其直径为．０５－４．５７ｍｍ。温度在２７．５－３１℃，幼鱼孵出需要２９个小时３０分钟。初孵仔鱼