人血小板生成素基因的克隆及CHO稳定细胞系的建立

 以人肝cDNA为模板克隆了人血小板生成素（Human Thrombopoietin,hTPO）基因,利用基因重组技术构建了带有新霉素抗性基因（neo）筛选标记的pcDNA3.1(+)-hTPO真核表达载体,将重组质粒瞬时转染293T细胞,用鼠抗人TPO 单抗Western blot 检测 TPO蛋白的瞬时表达;再将重组质粒转染中国仓鼠卵巢细胞(Chinese Hamster Ovary,CHO),应用400 μg/mL的G418筛选克隆,经PCR及Western blot验证,获得了3株hTPO蛋白表达水平不同的CHO细胞系,为获得大量蛋白并进行活性功能试验及临床应用奠定基础。     

黄瓜单体异附加系的筛选与观察

 将甜瓜属异源三倍体与栽培黄瓜‘北京截头’杂交，经胚胎拯救获得了两个2n=15 (14C+1H)的单体异附加系，编号分别为02—17和02—39。02一l7植株叶形为戟形，02—39为深陷的掌状形，并且果刺均为白色，不同于异源三倍体的掌状心脏形和黑刺，果形比异源三倍体黄瓜长而更接近普通黄瓜。细胞学研究发现，单体异附加系在终变期和中期I染色体构型主要是由7个二价体和1个单价体构成，出现三价体的频率为4％ ～5％ ，中期Ⅱ不均等分裂形成不同的两极，一极染色体为8条，另一极为7条。RAPD分析证明它们分刖附加了野生种Cucum／s hystrix的染色体。     

Doubled-haploid versus single-seed descent and S1-family variation for testcross performance in a maize population

Progress made in the in situ gynogenesis technique since 1990 now allows production of a high number of maize (Zea mays L.) doubled-haploid (DH) lines. The aim of the study was to compare DH lines versus selfing lines for testcross performance. DH and single-seed descent (SSD) lines were produced from random S₁ progenies of a broad-base population. For grain yield, kernel moisture, plant height, ear height and leaf length, the three population means were similar. Except for kernel moisture, the genetic variance of DH lines was nearly twice as high as the genetic variance of S₁ families, as expected. On the other hand, genetic variance among SSD lines was only 1.5 times higher than the genetic variance of S₁ families. This lower variance could be due to a selection bias in the method of production of SSD lines. However, for all traits, heritability of SSD or DH lines was higher than heritability of S₁ families. Epistasis effects in DH progenies were not significant. The consequence was a high correlation between S₁ testcross progenies and DH or SSD testcross progenies, meaning that the S₁ testcross value can be used to select the best families from which DH lines will be extracted. As a whole, the observed variation in DH lines appeared to be more in accordance with the observed variation among S₁ families than with the observed variation among SSD lines.     

Inhibition of telomerase in canine cancer cells following telomestatin treatment

Telomere shortening in normal somatic cells has been proposed as a major barrier to unlimited cellular proliferation. Telomerase is an enzyme capable of maintaining telomere length, and thus bypassing this barrier. In human beings, telomerase activity is restricted to cancer cells and cells of stem or germ cell lineages. Dogs represent a potentially useful clinical model for the development of telomerase‐based therapies because telomerase activity is also restricted to cancer cells and stem cells in this species. We examined the ability of telomestatin to inhibit telomerase activity in telomerase‐positive D17 and CMT7 canine cancer cell lines. At a concentration of 2 μM, telomestatin treatment resulted in a decrease in telomerase activity, telomere shortening, growth inhibition and apoptosis in telomerase‐positive cancer cells. These effects were not seen in telomerase‐negative skin fibroblasts or negative controls. These results confirm that telomestatin specifically inhibits telomerase activity in canine cancer cells and strengthens the usefulness of dogs as a model for testing telomerase‐based therapies.     

Inhibitory effects of LDL-ACM complex on subcutaneous implanted tumors in nude mice()

AIM: In order to evaluate the applicable value of LDL as a targeted vehicle for chemotherapeutic agents, we investigated and compared the inhibitory effects of LDL-ACM complex and free ACM on nude mice's subcutaneous implanted tumors derived from gastric cancer cell lines, SGC-7901 and NKM-45. METHODS: LDL-ACM complex was prepared and the tumor model of nude mice was established by subcutaneous implantation of SGC-7901 and NKM-45. Then, the groups of nude mice developed subcutaneous implanted tumors were received either LDL-ACM complex or free ACM. Subsequently, the tumor size, weight and leukemia cell counts were measured and the rates of tumor-inhibition and the survival were compared among the groups. RESULTS: The inhibitory effects of LDL-ACM complex on the tumors, especially on SGC-7901 implanted tumors were much more obvious than that of free ACM. It was also indicated that the action of LDL-ACM complex was mediated by LDL receptor. CONCLUSION: These results showed that LDL-ACM complex had significant inhibitory effects on the implanted tumors and the effect might be mediated by LDL receptor.     

天水旱作农业区膜侧小麦不同施肥水平增产效应分析

通过不同施肥条件对膜侧小麦水分利用、农艺性状、增产效应、经济效益等方面分析,研究干旱地区膜侧小麦的施肥问题。结果表明:施肥可以提高水分利用率1.1~3.1 kg/(mm.hm2),促进小麦生长发育;不同施肥水平对小麦的增产效应不同:低肥、中肥、高肥3种施肥水平分别比不施肥的对照增产731.5 kg/hm2,1461.0kg/hm2和1 648.5 kg/hm2,增产率分别为25.5%、50.9%和57.4%;经济效益以中肥水平(折合施纯N118.1kg/hm2,P2O594.5 kg/hm2和K2O 60.0 kg/hm2)最高,产值可达2 045.4元/hm2,产投比为2.7∶1。因此,在低、中、高3个施肥水平中,中等施肥最为有效、合理,有利于增产增收。     

Identification of QTLs related to sugar and organic acid composition in melon using near-isogenic lines

The soluble sugar and organic acid composition of melon (Cucumis melo L.) fruit flesh has become a major focus of plant breeding worldwide in an attempt to improve taste. Thus, sugar and organic acid profiles were characterized using near-isogenic lines (NILs) derived from the Spanish cultivar Piel de Sapo (PS) and the exotic Korean accession Shongwan Charmi (PI 161375). Fruits were cultivated in only one environment. These data were used to map 60 quantitative trait loci (QTLs): 18 for individual sugars, eight for sucrose equivalents, five for the glucose-to-fructose ratio, seven for the total sugar content and 21 for organic acids. Within the QTLs that were associated with the sugar profile, 27 defined the sugar content: eight for fructose, six for glucose, four for sucrose, and nine for sucrose equivalents. Although increased sweetness of selected NILs compared with the parental PS was achieved by an increased glucose or fructose content, only glucose heritability was above 0.5. A total of 21 QTLs (two with positive effects and nineteen with detrimental effects compared with the PS levels) controlled the organic acid profile: l-glutamic, ascorbic and succinic acids (the principal ones) and fumaric, citric, oxalacetic, and isocitric acids. The levels of sugars imparted by the PI 161375 introgression frequently decreased the score grades given to NILs by consumers. Within the 32 QTLs mapped for sensory traits, 27 were associated with lower scores in taste (nine QTLs), sweetness (eight QTLs) or global quality appreciation (nine QTLs); two with increased fruit sourness or sweetness and three with increased fruit bitterness. The QTLs defined herein may assist breeders to understand the overall organoleptic balance (sweetness, sourness, and umami taste) in melon fruit, particularly those located within linkage groups III, V, VI, and VIII to XI.     

The suitability of some blood gas and biochemical parameters as diagnostic tools or early indicators of ascites syndrome in broiler sire lines

In recent few years, there have been some attempts to find a reliable indicator trait as a selection criterion against susceptibility to ascites syndrome (AS). Blood parameters were of great interest as they could be measured in live animals without implementing an ascites‐inducing challenge (AIC). In this work, the suitability of some blood parameters was evaluated for diagnosing AS‐susceptible chicks in later steps of the disease in trial 1 as well as their early predictive ability in trial 2. In the first trial, one hundred 1‐day‐old chicks from two pure broiler lines namely S₁ and S₂ and, in the second trial, 226 1‐day‐old chicks from line S₂ were subjected to AIC. Saline drinking water (1200 mg/l) and lower‐than‐standard ambient temperatures were the implemented AICs in trials 1 and 2 respectively. The blood parameters including pH, partial pressure of O₂ (pO₂), partial pressure of CO₂ (pCO₂), bicarbonate ion concentration (BIC), percentage of haematocrit (HCT) and saturated haemoglobin (SaO₂) were measured twice per each bird at days 28 and 35 in trial 1 and once in trial 2 at day 21. The results of the first trial revealed that in line S₂ some of the blood parameters differed significantly between the ascitic and non‐ascitic groups following exposure to AIC. In this line, the incidence of AS was accompanied by a lower pO₂, SaO₂ and BIC, while with higher pCO₂ and HCT values. In the second trial, however, although almost all of the parameters showed meaningful differences between the ascitic and non‐ascitic broilers, only mean difference of BIC parameter was statistically significant. The general conclusion of this study is that the blood parameters can somewhat have diagnostic ability in the condition in which the AIC is already present, whereas the results did not approve their usefulness as early predictors of AS.     

鸡胚及禽类细胞系在生物医药领域的应用及研究进展

鸡胚因发育过程清楚,长久以来作为基础和应用科学研究领域重要的实验模型,尤其在鸡胚发育早期绒毛尿囊膜阶段,因其血管丰富,是天然的免疫缺陷宿主,是病理学、药理学和肿瘤学等研究领域的理想实验模型。作者简述了发育早期的鸡胚组织结构,并介绍了鸡胚绒毛尿囊膜在肿瘤研究、血管生成、器官移植、烧伤等疾病机理研究中的应用,以及在鸡胚病理模型基础上进行的抗肿瘤药物筛选的应用。重点介绍了鸡胚及禽类细胞系在病毒繁殖和疫苗生产、治疗性蛋白和单抗生产方面的应用研究进展。多种人源病毒、禽源病毒、支原体等可在鸡胚及禽类细胞上增殖,并用于疫苗生产。作者对常用的禽类纤维原细胞和多能干细胞的发展和特点进行了阐述,并总结了商业化的禽类细胞系来源以及部分易感病毒。鸡胚表达系统能够在目的蛋白特定位点产生人源化糖基,减少目的蛋白对人的过敏反应,且禽蛋廉价易得,可作为生产人用单克隆抗体和治疗性蛋白的合适供体。作者介绍了鸡胚及禽类细胞系在生物医药领域应用的最新进展,并对鸡胚作为动物模型在未来的应用进行了展望。     

不同玉米自交系耐寒性评价及差异分析

在室内低温条件下,对44份玉米自交系进行发芽期性状鉴定,并通过隶属函数法进行耐寒性综合评价,根据评价结果对耐寒玉米自交系和寒敏感玉米自交系进行苗期生长和生理生化特性分析,探索不同玉米自交系耐寒性差异生理机理。结果表明,利用发芽期发芽率、胚芽长、胚根长、根数为隶属函数法综合评价指标筛选出了3份强耐寒玉米自交系,包括玉米自交系Va35-2、Va102、ND246,中等耐寒玉米自交系1份,耐寒玉米自交系2份,寒敏感自交系16份,高寒敏感自交系22份。另外,结果显示随着温度逐渐降低(25℃、15℃、10℃、6℃),不同玉米自交系的发芽率、胚芽长、胚根长和根数呈下降趋势,但下降幅度不同,强耐寒玉米自交系各项指标下降的速度较慢;但是,从15℃低温开始,高寒敏感自交系的各项发芽指标下降很快,甚至为0。根据耐寒性综合评价结果,分别选取高耐寒玉米自交系Va35-2、Va102、ND246和高寒敏感玉米自交系K22、B68和H105W进行苗期耐寒性差异分析。6℃低温胁迫处理0、2、4、6 d后,高耐寒自交系Va35-2、Va102、ND246和高敏感自交系K22、B68和H105W的生长和生理生化指标都有不同程度的变化,但是Va35-2、Va102和ND246各项指标变化幅度较小,而K22、B68和H105W的变化幅度较大,不同品种间差异显著。总体来看,随着低温胁迫时间的延长,生长指标(株高、根长、地上部鲜重、地上部干重、地下部鲜重和地下部干重)和生理生化指标(相对含水量、脯氨酸、可溶性糖含量和保护酶活性(SOD、POD、CAT))呈下降趋势,高耐寒玉米自交系下降幅度较小,而高寒敏感玉米自交系下降幅度较大。相对电导率和丙二醛含量呈上升趋势,耐寒玉米自交系上升幅度较小,而寒敏感玉米自交系上升幅度较大。苗期性状的相关性分析表明各项生长和生理指标之间联系紧密、互相影响,在低温胁迫条件下调节植物的生长。