小鼠胎肝细胞透明质酸3D培养体系的建立

目的 利用透明质酸建立小鼠胎肝细胞3D培养体系。 方法 分离获得胚胎12-14天胎肝细胞,利用KM培养基进行初步2D肝干/祖细胞的筛选培养,并利用透明质酸及KM培养基配制水凝胶建立3D细胞培养体系。 结果 胎肝细胞在2D体系中呈现克隆状生长。分离培养获得的肝干/祖细胞克隆在透明质酸建立的3D培养体系保持增殖活性,并进一步获得肝细胞功能特性,表现为3D培养上清中白蛋白合成和尿素水平显著增加。Q-PCR结果显示随着3D培养时间的延长,其肝细胞干性标志如AFP、CK19、EpCAM、Prox1等表达水平都大幅度降低且接近成年小鼠肝脏表达水平。 结论 本研究成功建立基于透明质酸的小鼠胎肝细胞的3D无血清培养体系,并可促进小鼠胎肝细胞肝细胞功能进一步成熟。     

The use of spun columns in the purification of double-stranded cDNA appropriate for tailing

A reliable estimation of the efficiency of adding homopolymeric tails to double-stranded cDNA molecules by terminal deoxynucleotidyl transferase is extremely important in the construction of cDNA libraries. Appreciable differences in transformation efficiency result when the homopolymer tails to be annealed are of inadequate length. We report here that the use of a Sephadex G-50 spun column to remove oligo(dT)12-18 frequently coprecipitating in ethanol with the cDNA results in a more dependable estimation of tail lengths.     

北京地区红隼的迁徙路线和活动区域

红隼(Falco tinnunculus)被列为国家二级重点保护野生动物,是能同时适应农村和城市环境的小型猛禽,对维持城市生态系统稳定具有重要意义。2022年4月至7月,为在北京救助的7只红隼佩戴了卫星追踪器,追踪其活动轨迹,依据追踪的动物活动位点数据,采用净平方位移-时间曲线依次对各红隼的迁徙模式进行了判别,深入分析了迁徙红隼的迁徙时间、距离和路线等,并采用核心密度法分别计算了迁徙及留居型红隼95%及50%活动区面积。研究结果表明,在北京地区红隼的迁徙模式为部分迁徙,追踪的7只红隼个体(N01 ~ N07)中,4只为留鸟,1只为迁徙鸟,2只居留类型无法准确判断。N01为迁徙红隼,其度夏地和越冬地分别在内蒙古锡林郭勒盟和河北廊坊,此红隼秋季迁徙速度明显高于春季,其春季迁徙距离551 km,历时25 d,平均迁徙速度为22 km/d,而秋季迁徙距离412 km,历时2 d,平均迁徙速度为203 km/d,河北承德滦平县是其春季迁徙的重要中途停歇地。不同红隼个体间95%及50%活动区面积均存在较大差异,迁徙红隼N01 95%、50%活动区面积在度夏区分别为93.10 km²、17.50 km²,在越冬区分别为7.03 km²、0.99 km²;留居型红隼95%、50%活动区面积均值分别为1 165.34 km²、178.71 km²(n = 4),其中最大95%、50%活动区面积分别为4 320.26 km²(N02)、648.22 km²(N02),最小95%、50%活动区面积分别为2.80 km²(N03)、0.29 km²(N03)。本研究揭示了北京地区红隼的迁徙模式、迁徙路线、重要停歇地及活动区状况,为红隼的针对性保护和管理提供了科学依据。     

中国东、南部的3个腔蚓属蚯蚓新物种

本文报道了寡毛纲(Oligochaeta)巨蚓科(Megascolecidae)腔蚓属(Metaphire)3个新发现物种,分别是象头山腔蚓(M. xiangtoumontis Dong & Jiang sp. nov.),韩摆渡腔蚓(M. hanbaiduensis Dong & Sun sp. nov.)和长白山腔蚓(M. changbaimontis Dong & Shen sp. nov.)。象头山腔蚓受精囊孔2对,位于7/8 ~ 8/9节间,属于M. insulana物种群。韩摆渡腔蚓受精囊孔3对,位于6/7 ~ 8/9节间,属于M. houlleti物种群。长白山腔蚓受精囊孔2对,位于6/7 ~ 7/8节间,属于M. glandularis物种群。所有新物种均附有形态学描述、图片、与相似物种的形态学比较及与GenBank上亲缘关系相近物种的遗传距离计算分析。以上结果丰富了我国腔蚓属蚯蚓的物种多样性,并首次记录了采集于长白山国家级自然保护区的巨蚓科蚯蚓新物种。     

荷叶铁线蕨自然居群的遗传多样性研究

荷叶铁线蕨(Adiantumreniformevar. sinense)为我国特有植物,具有重要的经济价值,目前仅分布于重庆的少数地区。近几十年来,由于过度开发,该种的分布范围日益缩小,已处于灭绝的边缘。本研究利用等位酶标记检测了荷叶铁线蕨 6个自然居群共 136个个体的遗传多样性,共检测到了 5个酶系统的 14个位点,获得了 7个多态位点。结果表明:与其他蕨类植物相比,荷叶铁线蕨居群内的遗传多样性水平比较低。平均每位点的有效等位基因数(Ae)为 1. 778,多态位点百分率(P)为 0. 441,期望杂合度(He)为 0. 199,观察杂合度 (Ho)为 0. 235。其居群间的遗传分化也很低,居群间的遗传变异仅占总变异的 1. 49%,而 98. 51%的变异存在于居群内部。采用Hardy Wein berg平衡和固定指数F对荷叶铁线蕨的居群遗传结构进行了分析,结果表明其种群可能是以配子体间异交为主的混合交配体系。导致荷叶铁线蕨濒危的主要原因是生境的破坏以及过度开采所导致的生境片断化使其居群变小、近交率加大、遗传变异趋低,降低了其生存以及进化的潜力。     

The expression of a novel estrogen receptor, GPR30, in epithelial ovarian carcinoma and its correlation with MMP-9

The aim of the present study is to investigate the expression of a novel estrogen receptor, G protein-coupled receptor 30 (GPR30) and its correlation with matrix metalloproteinases-9 (MMP-9) in epithelial ovarian cancer (EOC). Ovary tissues were obtained from 39 female patients, including 30 cases of EOC and 9 cases of benign ovarian tumor. Four normal ovary tissues were used as control. Immunohistochemical staining was used to detect the expressions of GPR30 and MMP-9. Chi square test, Fisher's exact test and Spearman's rank correlation analysis were used for statistical analysis. The results showed that GPR30 overexpression rate in EOC cases was significantly higher than those in benign ovarian tumor and normal ovary cases. Whereas MMP-9 overexpression rate in EOC cases was significantly higher than that in normal ovary cases, without any difference to that in benign ovarian tumor cases. To demonstrate the relationship between GPR30 and clinicopathological variables of EOC, we further analyzed the pathology type, FIGO stage and age of patients sampled in our study. The analysis showed there were significant differences of GPR30 overexpression rate among various pathology types and different FIGO stages (P<0.05), and no significant difference of both GPR30 and MMP-9 among three age groups (P>0.05). Moreover, GPR30 expression was positively correlated with MMP-9 (r(s)=1.000, P=0.002). These results suggest that GPR30 may be involved in the invasion and metastasis of EOC, being a potential index of EOC early diagnosis and malignancy grade prediction.     

A sensitive signal-on assay for MTase activity based on methylation-responsive hairpin-capture DNA probe

This work develops a simple, sensitive and signal-on electrochemical sensor for methyltransferase (MTase) activity analysis. The sensor is composed of a methylene blue-modi?ed "signaling DNA probe" and a "capture DNA probe" tethered methylation-responsive hairpin DNA (hairpin-capture DNA probe). The thiol- modified hairpin-capture DNA probe at 5' end was firstly self-assembled on gold electrode via Au-S bonding. Methylation-induced scission of hairpin-capture DNA probe would displace the hairpin section and remain the "capture DNA probe" section on the gold electrode. Subsequently, the remained "capture DNA probe" on the gold electrode can hybridize with the methylene blue-modi?ed "signaling DNA probe", mediating methylene blue onto the gold electrode surface to generate redox current. It was eT on state. The developed facile signal-on electrochemical sensing system showed a linear response to concentration of Dam MTase range from 0.1 to 1.0 U/mL. The detection limit of Dam MTase activity was determined to be 0.07 U/mL and the total detection time is 7h. The sensor also has the ability to provide information about the dynamics of methylation process. Furthermore, we demonstrated that this sensor could be utilized to screen inhibitors or drugs for Dam MTase.     

基于MuSIASEM理论的大连经济技术开发区可持续发展评价

基于社会生态系统代谢多尺度综合评估（MuSIASEM）理论,对2000-2007年大连经济技术开发区人类活动时间、体外能投入量和增加值进行综合评价.结果表明：2000-2007年,大连开发区居民生活水平逐年提高;农业逐渐萎缩;第二产业仍为区域支柱产业,能耗较大;第三产业发展落后于第二产业;区域整体和各个产业的体外能代谢率明显提高,能源密度不断下降;人类活动时间与体外能投入量不断减少的情况下,经济增加值稳步增长,区域整体发展呈可持续态势.     

Spindlin1, a novel nuclear protein with a role in the transformation of NIH3T3 cells

spindlin1, a novel human gene recently isolated by our laboratory, is highly homologous to mouse spindlin gene. In this study, we cloned cDNA full-length of this novel gene and send it to GenBank database as spindlin1 (Homo sapiens spindlin1) with Accession No. AF317228. In order to investigate the function of spindlin1, we studied further the subcellular localization of Spindlin1 protein and the effects of spindlin1 overexpression in NIH3T3 cells. The results showed that the fusion protein pEGFP-N1-spindlin1 was located in the nucleus and the C-terminal is correlated with nuclear localization of Spindlin1 protein. NIH3T3 cells which could stably express spindlin1 as a result of RT-PCR analysis compared with the control cells displayed a complete morphological change; made cell growth faster; and increased the percentage of cells in G2/M and S phase. Furthermore, overexpressed spindlin1 cells formed colonies in soft agar in vitro and formed tumors in nude mice. Our findings provide direct evidence that spindlin1 gene may contribute to tumorigenesis.