胚胎冷冻技术及其在转基因小鼠中的应用

随着生物工程技术的发展,出现了许多生物工程小鼠品系,促使胚胎冷冻技术得到了广泛应用。胚胎冷冻技术不仅为转基因动物提供可用胚胎,还使胚胎移植不受时间、地域限制,简化引种过程,防止疾病传播,为建立优良动物的胚胎库提供了条件。综述了几种常用的胚胎冷冻方法及其在转基因小鼠中的应用研究,指出了胚胎冷冻方法存在的主要问题,并展望了其发展前景。     

Meiotic Response of In vitro Matured Canine Oocytes under Different Proteins and Heterologous Hormone Supplementation

The impact of TCM‐199 supplemented with different proteins and heterologous hormones on the in vitro maturation (IVM) rate of bitch oocytes was evaluated by nuclear staining under fluorescence microscopy. Oocytes were recovered by slicing of ovaries from bitches presented at various stages of oestrous cycle to ovariohysterectomy. The basic culture medium was TCM‐199 supplemented with 25 mM Hepes/l, with 10% heat‐inactivated oestrous cow serum (ECS), 50 μg/ml gentamicin, 2.2 mg/ml sodium bicarbonate and 22‐μg/ml pyruvic acid, 1.0‐μg/ml oestradiol (E 8875; Sigma), 0.5‐μg/ml follicle‐stimulating hormone (FSH) (Folltropin‐V; Vetrepharm Inc., Ontario, Canada) and 0.03 IU/ml human gonadotropin (hCG) (Profasi HP; Serono, Aubonne, Switzerland). Oocytes were distributed randomly between basic culture medium (control) and the corresponding experimental treatment. Hormone treatments were: oocytes cultured in; (1) medium without FSH, (2) control medium supplemented with 20 μg/ml oestradiol, or (3) medium supplemented with 1 μg/ml human somatotropin (hST; Humatrope, Lilly, Saint Cloud, France). The second experiment consisted of oocytes cultured in medium supplemented with 0.4% (w/v) bovine serum albumin (BSA, fraction V; Gibco Grand Island, NY, USA) instead of ECS, or oocytes cultured in medium with 10% inactivated oestrous bitch serum (EBS) instead of ECS. Oocytes were cultured in 100 μl droplets (up to 25 oocytes per drop) under mineral oil at 37°C in a 100% humidified atmosphere containing 5% CO₂ in air. After 72 h of IVM, the highest rates (p < 0.05) of meiotic resumption were achieved with the 0.4% BSA supplementation. A positive influence on the metaphase II (MII) acquisition rate was observed with hST supplement. Oocytes cultured with 10% EBS supplementation did not develop to the MII stage. The results in this study show that the protein and hormone supplements to TCM‐199 culture medium tested did not promote the final steps of IVM of bitch oocytes.     

X-Chromosomal Monosomy (77, XO) in a Doberman Pinscher With Gonadal Dysgenesis

A 6-month-old stunted female Doberman Pinscher was found to have a 77,X0 chromosomal complement. The ovaries were small, consisting primarily of interstitial-type cells and solid epithelial cords. The dam, sire, and a male littermate had normal karyotypes.     

Comparison of the properties of hyaluronate products used in treatment of joint dysfunction in horses

A standardized investigation was conducted to compare the biophysical properties of hyaluronate products used in the treatment of joint dysfunction in horses. Results showed that two of the five products, Hyvisc® and Hylartin®V, contained higher-weight molecular hyaluronate (greater than 1.5×10⁶). Other investigations have shown that higher molecular weight hyaluronate is effective in providing physical resistance to shear, in stimulating endogenous hyaluronate synthesis, and in inhibiting excess cell proliferation which may be associated with synovial pannus formation. It is suggested that the therapeutic benefits of hyaluronate products may be dependent on the molecular size of the hyaluronate molecules. Higher molecular weight hyaluronate products would be expected to be more effective than lower molecular hyaluronate products in providing therapeutic benefits.     

外源物质对辣椒种子发芽的影响研究

用一定浓度的赤霉素(GA3)、生长素(IAA)、过氧化氢(H2O2)、硝酸钾(KNO3)、萘乙酸(NAA)溶液处理可显著提高辣椒砧木品种肩车种子的发芽率,并对其发芽势、发芽指数有一定的促进作用。比较几种药剂对辣椒种子发芽的影响发现,GA3和IAA对辣椒种子的发芽影响效果最佳,最适宜浓度分别为200 mg/L和1.5 mg/L。     

冬枣树皮疱斑病防治药剂筛选试验

冬枣树皮疱斑病Phoma persicae是近年在冬枣上新发生的一种重要病害,2009～2011年对其开展了室内抑菌测定和田间防效试验。结果表明,室内抑菌效果较好的药剂为苯醚甲环唑、丙环唑、氟硅唑、苯醚甲环唑.丙环唑、恶唑菌酮.氟硅唑,抑菌率分别为98.7%、96.2%、94.9%、93.6%、92.3%;田间防治效果较好的药剂有苯醚甲环唑、嘧菌酯、氟硅唑、中生菌素,防治效果分别为90.2%、87.1%、80.4%、69.1%,可以在防治冬枣树皮疱斑病生产中推广使用。     

用荷斯坦奶牛性控精液生产体内性控胚胎

采用荷斯坦奶牛X性控冻精和超数排卵技术结合生产奶牛性控胚胎,以建立高效、低成本的体内性控胚胎生产方法。研究发现,使用CIDR和Cue-Mate进行奶牛同期发情处理,发情率分别为90.9%和100%(P<0.05),且Cue-Mate的发情时间更加集中;加拿大的FOLLTROPIN-V和中国科学院动物研究所的FSH对奶牛超排效果影响不显著,分别为头均胚胎数10.08枚±8.08枚、头均可用胚胎数2.38枚±3.07枚和头均胚胎数6.57枚±4.31枚、头均可用胚胎数2.00枚±1.88枚,无显著差异(P>0.05);奶牛发情后12h～14h和16h～19h人工授精,分别获得头均胚胎数10.25枚±5.53枚、头均可用胚胎数5.33枚±4.52枚和头均胚胎数10.13枚±6.93枚、头均可用胚胎数5.88枚±6.26枚,无显著差异(P>0.05)。但是未受精胚胎数差异显著(27对7,P<0.05)。     

济源市森林城市景观格局分析

以济源市2007年森林资源调查数据和2008年绿化数据以及卫星遥感图、城市绿地现状图等相关信息作为数据源,选取多样性指数、景观均匀度指数、城市绿地破碎度指数等3个景观生态格局评价指标,对济源市城区森林和市域森林景观格局进行了分析。结果表明：济源市城区森林绿地景观的均匀度指数较低,景观多样性整体程度有待进一步提高;市域森林景观格局分布不均匀,少数森林景观类型控制着整个景观格局。