猪体细胞核移植电融合参数的研究

试验旨在研究不同电融合参数(场强、脉冲时程)对猪核移植胚融合率及发育能力的影响。采用44～48h成熟培养的去核猪卵母细胞作为受体细胞,颗粒细胞为供体。结果表明,在20μs脉冲时程条件下,0.8kV·cm-1组融合率显著高于0.4、1.2、1.6kV·cm-1组(P0.05)。0.8、1.2kV·cm-1组卵裂率显著高于0.4、1.6kV·cm-1组(P0.05)。场强为0.8kV·cm-1条件下,20μs组融合率显著高于30、40、50μs组(P0.05)。卵裂率20、30μs组显著高于40、50μs组。根据试验数据,本试验室最佳融合电参数为:0.8kV·cm-1、20μs,1次脉冲。     

用单层分化法进行胚胎干细胞神经分化的研究

用单层分化法研究了 ES细胞从 0～ 316 h(第 14天 )的神经分化过程。无饲养层培养的 ES细胞在无血清的N2 B2 7培养基中开始神经分化。从 72 h开始 ,神经外胚层的特异性转录因子 Sox1表达 ,在克隆的边缘逐渐出现向外迁移的细胞 ,形状呈圆形或椭圆形 ,有 1～ 2个细胞突起。在 191h(第 8天 )之后 ,细胞可迁移至克隆边缘的 2 0 0 μm之外 ,这些有细长突起的细胞表达神经元特异性蛋白 Tau。以后 ,迁移细胞的突起开始交叠成网状。 316 h(第 14天 )时 ,细长的纤维可跨越在不同的克隆之间 ,长达上千微米。此时 ,免疫染色的结果表明 ,在克隆的中央有较多的圆形 Nestin阳性细胞 ,而克隆的边缘有大量的 β- tubulin 阳性细胞 ,它们交织成复杂的网状图像。阶段性表达基因的表达时序研究表明 ,ES细胞的特异性转录因子 Oct4开始减弱时 ,Sox1表达开始 ,此时细胞由全能状态进入早期神经分化状态 ;Tau的表达是 Sox1表达的下游事件。这一时序与胚胎发育中神经发生的时序相符。单层分化法能直观地展示 ES细胞的神经分化过程 ,可作为研究细胞分化、胚胎发育及药物筛选的体外模型     

Record keeping,genetic selection,educational experience and farm management effects on average milk yield per cow,milk fat percentage,bacterial score and bulk tank somatic cell count of dairy farms in the Central region of Thailand

A study was conducted to estimate the record keeping, genetic selection, educational, and farm management effects on average milk yield per cow (AYC), milk fat percentage, bacterial score, and bulk tank somatic cell count (BTSCC) of dairy farms in the central region of Thailand. Farms were located in the provinces of Saraburi and Nakhon Ratchisima and were members of the Muaklek dairy cooperative. Records from individual animals were unavailable. Thus, farm records of milk yield, milk fat percentage, bacterial score, and BTCCC were collected from July 1, 2003 through June 30, 2006. Additional record keeping, genetic selection, education, and farm management information was collected through a questionnaire in May of 2006. Data from the Muaklek dairy cooperative and the questionnaire were then merged by a farm identification number. A single trait mixed model was used to analyze AYC, milk fat percentage, and BTSCC, while a log linear model was used to analyze bacterial score. Results showed that farms that kept records on individual animals had higher (P < 0.05) milk fat percentages and lower bacterial scores than farms that did not. Farms that used genetic information (EBV) and phenotypes when selecting sires were higher (P < 0.05) for milk fat percentage than farms that used only phenotypes and personal opinion. Farms milking cows with a single unit milking machine and by hand, had higher (P < 0.05) bacterial scores and BTSCC than farms using only a single or multi unit machine. Overall farms that kept individual animal records, used EBV when selecting sires, used a single method for collecting milk, and used family labor achieved higher performance from their herds than farms that did not.     

南方红豆衫细胞体系建立的优化

[目的]优化南方红豆杉愈伤组织诱导及训化条件。[方法]以南方红豆杉幼芽、幼茎以及幼叶为外植体,比较不同外植体诱导愈伤组织的效率,并对愈伤组织的增殖条件及悬浮细胞培养条件进行了研究。[结果]以幼芽作为外植体的诱导效果最佳;南方红豆杉外植体消毒的最佳方法为链霉素浸泡2 h＋洗洁精浸泡3 h＋75%酒精消毒30 s＋10%次氯酸钠浸泡25 min＋1‰氯化汞10 min;愈伤继代的最佳配方为B5＋4.0 mg/LNAA＋0.5 mg/L2.4-D＋0.2 mg/LGA＋0.5 mg/L6-BA＋2 g/L活性碳。[结论]该研究建立了南方红豆杉的高效再生体系。     

The second-generation curcumin analogue RL71 elicits G2/M cell cycle arrest and apoptosis in canine osteosarcoma cells

Canine osteosarcoma is an aggressive cancer, comprising 85% of canine bone neoplasms. Current treatment practices of surgery and chemotherapy increase 1-year survival by only 45%. The curcumin analogue RL71, has demonstrated potent in vitro and in vivo efficacy in several models of human breast cancer through increased apoptosis and cell cycle arrest. Thus, the present study aimed to investigate efficacy of curcumin analogues in two canine osteosarcoma cell lines. Osteosarcoma cell viability was assessed using the sulforhodamine B assay and mechanisms of action were determined by analysing the levels of cell cycle and apoptotic regulatory proteins via Western blotting. Further evidence was obtained using flow cytometry to detect cell cycle distribution and the number of apoptotic cells. RL71 was the most potent curcumin analogue with EC₅₀ values of 0.64 ± 0.04 and 0.38 ± 0.009 μM (n = 3) in D-17 (commercial) and Gracie canine osteosarcoma cells, respectively. RL71 significantly increased the ratio of cleaved-caspase 3 to pro-caspase 3 and the level of apoptotic cells at the 2× and 5× EC₅₀ concentration (p < 0.001, n = 3). Furthermore, at the same concentration, RL71 significantly increased the number of cells in the G2/M phase. In conclusion, RL71 has potent cytotoxic activity in canine osteosarcoma cells triggering G2/M arrest and apoptosis at concentrations achievable in vivo. Future research should further investigate molecular mechanisms for these changes in other canine osteosarcoma cell lines prior to in vivo investigation.     

人乳腺癌细胞、结肠癌细胞和宫颈癌细胞生长曲线测定

为检测比较人乳腺细胞(MDA-MB-231、MCF-7、HBL-100)、人结肠癌细胞(HCT-8)和人宫颈癌细胞(HeLa)生长曲线的差异性.本实验首先常规培养MDA-MB-231、MCF-7、HBL-100、HCT-8和HeLa细胞,然后应用MTT方法测定这几种细胞的生长曲线.在本实验条件下,这几种细胞生长速度从高...     

A Cathepsin B Inhibitor,E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos

Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.     

管状分子分化离体培养系统的建立及影响因素

管状分子是植物木质部的主要组成成分,具有复杂的形态学特征,在植物的生长发育过程中发挥重要作用。在离体培养条件下,植物细胞可以通过分化或转分化的方式形成管状分子,为此建立了百日草、拟南芥、杨树等离体培养模式系统,用于观测管状分子分化细胞形态结构特征的变化以及相关影响因子的调节作用,并取得较大的进展。研究资料表明：离体条件下不同植物细胞分化形成管状分子培养条件不同,其中植物激素、木质形成素、渗透压、Ca2+、起始细胞密度、pH等均参与管状分子的分化调控。迄今为止,百日草系统是研究管状分子分化较为成熟的系统,而杨树系统是研究管状分子分化最具前景的林木模式系统。这些模式系统的建立可为单细胞水平上研究管状分子的分化提供极好的平台,从而可在分子水平上认识细胞分化进程及其调节机制,最终达到木材改良的目的。     

三种有机磷农药对大麦毒性效应的研究

使用浸种和浸种加喷施两种方法,研究了3种有机磷农药———马拉硫磷、对硫磷和久效磷在不同浓度条件下对大麦幼苗生长发育、根细胞分裂及叶绿素含量的影响。结果表明,3种有机磷农药对大麦种子萌发和根伸长均有一定的抑制作用,抑制程度随浓度增大而加强。幼苗根伸长比种子萌发对有机磷农药胁迫更为敏感。不同浓度的有机磷农药处理后均能抑制根细胞分裂,使有丝分裂指数下降。浸种法和浸种加喷施法都能使大麦叶片叶绿素含量降低,但浸种加喷施法对叶绿素合成的影响明显大于浸种法。3种有机磷农药对大麦幼苗的毒性程度依次为马拉硫磷>对硫磷>久效磷。     

昆明地区四种蛙血红细胞微核与环境质量关系

用采自昆明金殿—世博园周围、松花坝水库、西郊花红洞、马街和安宁温泉5个点的滇蛙、昭觉林蛙、华西蟾蜍和大蹊铃蟾的红细胞微核实验检测采样点水样的致突变性。研究结果表明,同一采集点中,不同种类蛙对相同环境污染的敏感性不同。滇蛙对环境的敏感性较高。滇蛙较其他种类分布广、数量多,易于采集,因此它可以作为监测水体污染较为理想的指示生物。对不同采集点蛙的微核率差异显著性(x^2)检验表明,昆明金殿—世博园和花红洞的生境质量较好。森林、农田和鱼塘3种不同小生境中几种蛙的平均微核率分别为1．06‰、4．26‰和4．80‰,该结果与3种生境污染程度的高低有关。最后就各生境采集到的蛙的种类、数量及生境状况等综合指标探讨了昆明地区的环境质量。