牛MEN1基因真核表达载体的构建及其表达分析

多发性内分泌腺瘤致病因子1 (multiple endocrine neoplasia Ⅰ,MEN1)参与乳腺发育与泌乳行为的调控.本研究克隆了牛(Bos taurus) MEN1基因(bMEN1)的全长cDNA,并在不同细胞中检测bMEN1mRNA及其编码蛋白menin的表达情况.根据GenBank中bMEN1基因序列,设计特异性引物,用qRT-PCR方法得到附带EcoR Ⅰ和HindⅢ酶切位点的bMEN1片段,并将其克隆到真核表达载体pcDNA3.1-myc-his(A)中.体外转染物种同源性细胞牛乳腺上皮细胞(bovine mammary gland epithelial cell,MAC-T)和异源性细胞中国仓鼠(Cricetulus barabensis)卵巢细胞(Chinese hamster ovary cells,CHO)、小鼠(Mus musculus)成肌细胞(mouse myoblast cells,C2C12),利用qRT-PCR和Westem blot技术检测转染前后bMEN1 mRNA和蛋白menin的表达.双酶切和基因测序结果表明,成功构建了bMEN1基因的真核表达载体pcDNA3.1-myc-his-bMEN1.所建立的转染体系可以使目的基因bMEN1在3种不同细胞中成功表达mRNA和目的蛋白,转染后24 h都达到最高表达量,并达到极显著水平(P＜0.01),之后逐渐降低.其中,CHO细胞中的转染24 h后bMEN1 mRNA和menin蛋白的表达量分别是对照的28 415倍和5.65倍.本研究建立了bMEN1基因的真核表达载体及其转染体系,能够在不同细胞中成功表达,为体外研究MEN1基因对于乳腺的调节功能及其对于机体代谢的调节机制提供了一种工具和技术体系.     

激素对蝴蝶兰快速繁殖的影响

将蝴蝶兰原球茎在培养期间置于MS附加 6 BA ,NAA ,IBA ,2 .4 D等激素的培养基 ,诱导分化、增殖。结果表明 ,无菌母株在MS附加高浓度 6 BA(5 .0mg·L-1)和NAA(2 .5mg·L-1)条件下培养 2 5～ 30d后 ,转接到单独使用 6 BA(2 .0mg·L-1)的培养基中 ,并继代 2～ 3次 ,可连续不断地分化、增殖原球茎 ;若 6 BA浓度继续下调并与适当浓度NAA相配合 ,则有利于原球茎生长 ;低浓度 6 BA与IBA配合使用有利于分化苗生根。     

酮病对泌乳早期乳牛体内代谢和内分泌的影响

随机选取某奶牛场10头患临床酮病乳牛和10头同期健康对照组乳牛，检测了2组乳牛血液10项指标，阐明了酮病对泌乳早期乳牛体内代谢和内分泌的影响。结果显示，酮病乳牛血糖浓度极显著降低（P〈O．01），血浆NEFA和BHBA的浓度明显增高（P〈O．01）；酮病乳牛血浆Ins、LP、NPY、E2的浓度和Ins／Gn比值均明显降低（P〈O．05），而血浆Gn浓度未明显升高，P4未明显降低（P〉0．05）。表明，酮病乳牛体内某些激素协调作用紊乱会妨碍酮病乳牛能量负平衡的缓解，并将对产后生殖机能产生不良影响。     

Evaluation of success rate of laparoscopic castration without orchidectomy in 32 mature horses

The surgical success of laparoscopic castration without orchidectomy results from avascular necrosis of the testes. However, failures and subsequent production of testosterone and stallion‐like behaviour have been previously identified. Laparoscopic castration without removal of the testes was performed in 32 horses with 2 normal descended scrotal testes between July 2006 and October 2012. The objectives of our study were to evaluate the success rate of laparoscopic castration without orchidectomy on descended testes in our population and assess complications and recovery time. Endocrine tests were performed after surgery and owners also asked to report on their horse's behaviour and ability to resume exercise. Castration was deemed successful if endocrine test results were in accordance with gelding values. Failed horses were castrated using a conventional technique and histology performed when possible. Basal testosterone levels decreased below 3 nmol/l after surgery in 30/32 cases. However, in 2 horses, stallion behaviour persisted and histological evaluation after inguinal castration showed residual viable tissue. Two horses maintained high testosterone levels after surgery associated with stallion‐like behaviour. Based on these results, 12.5% of horses (4/32 horses) failed to be considered a gelding based on either endocrine tests and/or histology. All horses, with one exception, were able to resume exercise less than a week after the procedure. Laparoscopic castration without orchidectomy appears to be a safe procedure. When assessing the success of laparoscopic castration, our total failure rate was 12.5% (4/32 horses) which is significantly higher than previously reported.     

Pollution of Environmental Endocrine Disrupting Chemicals(EDCs) in Water and Its Adverse Reproductive Effect on Fish

越来越多的环境内分泌干扰物（ EDCs）不断释放到环境,并通过大气沉降、地表径流、土壤淋溶和直接排放等方式进入水体,从而使水体成为 EDCs 存在的主要场所之一。为引起社会对水体 EDCs 污染的广泛关注,并积极采取EDCs 危害防治措施,保护鱼类资源和水生态系统,本文介绍了 EDCs 的分类,详述了自然水体、饮用水源水以及自来水中EDCs 污染情况,阐明了 EDCs 对鱼类的生殖危害。资料分析显示,EDCs 能够扰乱生物体内分泌功能,导致生殖器官、生殖机能和生殖行为异常,引起生育力下降,甚至生物繁殖机能损害,并最终导致种群数量下降,以至物种灭绝。虽然国内外已经开展了一些关于 EDCs 对生物危害等方面的研究,但大都处于起步阶段,存在着基础研究薄弱、识别和鉴定困难或代价太大等问题,有关工作亟需全面、深入开展。     

Endocrine and gonadal changes during the annual reproductive cycle of the freshwater teleost,Stizostedion vitreum

The annual reproductive cycle of walleye (Stizostedion vitreum) was characterized by documenting changes in gonadal development and serum levels of estradiol-17β (E₂), testosterone (T), 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P), and 11-ketotestosterone (11-KT) in wild fish captured from upper midwestern lakes and rivers throughout the year. Fish from the populations used in this study spawn annually in early- to mid-April. Walleye showed group synchronous ovarian development with exogenous vitellogenesis beginning in autumn. Oocyte diameters increased rapidly from ∼ 200 μm in October to ∼ 1,000 μm in November, and reached a maximum of 1,500 μm just prior to spawning. Changes in gonadosomatic indices (GSIs) paralleled changes in oocyte diameters. Serum E₂ levels in females increased rapidly from low values in October (< 0.1 ng ml⁻¹) to peak levels of 3.7 ng ml⁻¹ in November, coinciding with the period of the most rapid ovarian growth. Subsequently, E₂ levels decreased from December through spawning. Serum T levels exhibited a bimodal pattern, increasing to 1.6 ng ml⁻¹ in November, and peaking again at 3.3 ng ml⁻¹ just prior to spawning. We detected 11-KT in the serum of some females at concentrations up to 5.6 ng ml⁻¹, but no seasonal pattern was apparent. In this study (unlike our results in a related study) 17,20-P was not detected. In males, differentiation of spermatogonia began in late August, and by January the testes were filled (> 95% of germ cells) with spermatozoa. Mature spermatozoa could be expressed from males from January through April. GSIs ranged from 0.2% (post-spawn) to 3.2% (pre-spawn). Serum T levels rose from undetectable levels in post-spawn males to 1.6 ng ml⁻¹ by November, remained elevated throughout the winter, and peaked at 2.8 ng ml⁻¹ I prior to spawning. Levels of 11-KT in males remained low (< 10 ng ml⁻¹, from post-spawning through January, then increased significantly by March and peaked just prior to spawning at 39.7 ng ml⁻¹. Our results indicate that vitellogenesis and spermatogenesis are complete or nearly so, in walleye by early winter, and suggest that it may be possible to induce spawning in this species several months prior to the normal spawning season by subjecting fish to relatively simple environmental and hormonal treatments.     

昌潍白猪内分泌腺的生长发育规律

系统研究了1～8月龄昌潍白猪(192头)内分泌腺的性状指标,并由此得出了其内分泌腺的生长发育规律及其与体重变化的回归方程.结果表明,肾上腺在1～3、5～6月龄生长速度最快(P＜0.01);甲状腺在1～8月龄有2个生长高峰期,即4月龄和6月龄;胸腺的生长高峰期集中于3～4月龄(P＜0.01),颈部胸腺在1～3月龄生长速度显著快于胸部胸腺,胸部胸腺左侧生长发育明显快于右侧;脑垂体在2～6月龄生长速度较快.肾上腺、甲状腺、胸腺、脑垂体随体重变化的回归方程分别是y=0.019 9x+0.801 6(r=0.975 9,P＜0.01);y=0.163 4x+1.286 8(r=0.976 1,P＜0.01);y=1.760 8x+26.995 0(r=0.946 2,P＜0.01);y=0.002 4x+0.062 0(r=0.908 3,P＜0.01).     

全蚕粉对鹌鹑生长、营养、免疫及内分泌的影响

在日粮中添加 1 g· kg- 1 全蚕粉对鹌鹑的试验结果表明 :1饲喂 1 0、1 5、2 1 d后 ,体重分别增加 5 .0 0 %、6.67%和 1 1 .1 2 % ;2饲喂 2 1 d后 ,试验组料重比低于对照组 ;3血清总蛋白、白蛋白含量分别降低 1 2 .76%和 2 0 .84% ,血清葡萄糖、总胆固醇含量及白蛋白 /总蛋白比值差异不显著 ,但血清α-氨基氮含量试验组提高 1 8.84% ;4小肠粘膜麦芽糖酶和蔗糖酶活力分别下降 2 8.5 0 %和 3 8.67% ;5 T淋巴细胞转化率试验组比对照组高 2 0 .87% ;6血清胰岛素样生长因子含量提高 1 2 .1 4% ,生长激素和甲状腺素 T4水平分别提高 3 0 .0 8%和 2 0 .81 % ,T3差异不显著     

日粮中添加大豆黄酮对大鼠生长性能和有关内分泌的影响

将 2 4只断乳雄性大鼠随机等分为实验组和对照组 ,实验组在基础日粮 (对照组 )中添加大豆黄酮 3mg·kg-1,持续 30d。结果表明 ,与对照组相比 ,实验组大鼠日增重和采食量分别提高 14 90 % (P <0 0 1)和 18 31% (P <0 0 5 ) ;胴体重、腰胁部脂肪重、腓肠肌重、股骨重分别提高 16 6 % (P <0 0 5 ) ,11 5 % ,8 8%和 14 5 % (P <0 0 5 ) ;骨密度、血钙分别提高 9 0 9%和 2 4 1% ,而碱性磷酸酶 (AKP)活性降低 14 6 1% (P <0 0 5 ) ;血液胰岛素样生长因子 I (IGF I)和睾酮含量分别提高 36 86 % (P <0 0 5 )和 17 30 % (P <0 0 5 ) ,而雌二醇含量降低 2 4 11% (P<0 0 1)。提示 :大豆黄酮能显著增强雄性动物的骨骼代谢 ,促进生长 ,并与性激素和生长激素分泌系统有密切关系     

影响分娩母猪母性行为因素的探讨

分娩母猪母性行为的好坏直接影响到仔猪的成活率,尤其是一些母性行为失常的母猪直接咬死、踩死、压死仔猪,给养猪业造成巨大的经济损失。母猪分娩是受多种因素如神经、内分泌和胎儿等多方面影响和调控的一个复杂过程。近几年,虽然许多专家对分娩的机理进行研究,但到目前为止对造成母猪母性行为失常原因及机理的了解还不是很清楚。就分娩环境、机体内分泌、母猪遗传背景及产仔经验等影响分娩母猪母性行为的4个主要因素进行综述和讨论。