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1.
旋毛虫肌幼虫可溶性抗原,排汇分泌抗原的电泳分析   总被引:2,自引:0,他引:2  
枉文报道了应用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),聚丙烯酰胺凝胶等电聚焦(IEF)电泳及二维电泳(IEF/SDS-PAGE)对旋毛虫肌幼虫排汇分泌(ES)抗原和肌幼虫可溶性抗原的分析结果。肌幼虫ES抗原经SDS-PAGE后用考马斯亮蓝染色蛋白质,结果显示16条蛋白带,分子量范围21 ̄80KD,其中主带9条。IEF电泳后分别用PAS染多糖、考马斯亮蓝R-250染蛋白质、Nile'  相似文献   

2.
弓形虫GJS株速殖子通过传代细胞(BHK-21)培养,制备排泄分泌抗原(E/SA),蛋白含量为2.095mg/ml;SDS-PAGE凝胶电泳分析显示21条区带,分子量在18~120KD,其中68KD为主带,次带7条。用E/SA与佐剂(M-206)制备E/SA疫苗,对30只(2公,28母)弓形虫血检抗体阴性,混群自然交配50天的适龄健康绵羊随机分组进行免疫,间隔20天再加强免疫1次。第1组于混群后8  相似文献   

3.
IGF-Ⅰ对山羊睾丸间质细胞分泌睾酮的调节机理   总被引:2,自引:0,他引:2  
以30~40日龄山羊的睾丸间质细胞为研究对象,采用体外细胞培养的方法,用胰岛素样生长因子-Ⅰ(IGF-Ⅰ)和hCG进行不同的处理,用放射免疫分析法(RIA)测定了睾酮和cAMP的含量,用荧光分光光度分析法测定了DNA含量。结果表明,IGF-Ⅰ能促进体外培养的山羊睾丸间质细胞cAMP生成、睾酮分泌及DNA合成,以上作用随IGF-Ⅰ剂量的增大而加强,并且与hCG有协同作用。根据以上结果首次提出IGF-Ⅰ调节山羊睾丸间质细胞分泌睾酮机理的假设:IGF-Ⅰ与其膜受体结合引起cAMP增多,cAMP通过激活一系列合成DNA和睾酮所需酶类而使DNA和睾酮的合成增加。这与催乳素(PRL)和白细胞介素-1α(IL-1α)等含氮激素(因子)调节间质细胞分泌睾酮的机理不同,而与FSH、LH等含氮激素调节性腺分泌类固醇激素的作用机理相似。  相似文献   

4.
旋毛虫肌幼虫可溶性抗原、排泄分泌抗原的电泳分析   总被引:2,自引:0,他引:2  
本文报道了应用十二烷基硫酸钠一聚丙烯酰胺凝胶电泳(SDS-PAGE),聚丙烯酰胺凝胶等电聚焦(IEF)电泳及二维电泳(IEF/SDS-PAGE)对旋毛虫肌动虫排泄分泌(ES)抗原和肌幼虫可溶性抗原的分析结果。肌幼虫ES抗原经SDS-PAGE后用考马斯亮蓝染色蛋白质,结果显示16条蛋白带,分子量范围21~80KD,其中主带9条。IEF电泳后分别用PAS染多糖、考马斯亮蓝R-250染蛋白质、Nile's蓝染脂、醋酸a-萘酯/坚固蓝染酪酶同工酶,结果肌幼虫ES抗原分别显示16,26、7及0条带;肌幼虫可溶性抗原分别显示21、38、4及11条带。二维电泳后用考马斯亮蓝G-250染色多肤斑点,结果ES抗原显示多肽斑点61个;肌幼虫可溶抗原显示122个多肽斑点。  相似文献   

5.
旋毛虫肌幼虫排泄-分泌(ES)抗原产来源于体外培养的旋毛虫肌幼虫的排泄-分泌物。为研制具有ES抗在功能的旋毛虫基因重组抗原,首先用AGPC法提取旋毛虫肌幼虫总RNA并进行变性琼脂糖凝胶电泳分析,发现它缺少真核生物所具有的28SrRNA。根据编码45000和49000蛋白的基因结构,设计并合成2对PCR引物,用RT-PCR方法分别扩增出了2个目的基因(TSPG和PSPGⅡ)。序列分析发现,TSPG在  相似文献   

6.
用几种电泳方法分析牦牛肝片吸虫不同部位抗原   总被引:1,自引:0,他引:1  
乔代蓉  赵坚 《四川草原》1997,(2):48-51,64
应用SDS-PAGE、等电聚焦电泳(IEF)及双向电泳三种电泳方法分析牦牛肝片吸虫成虫四种抗原(头抗原-HA、体抗原-BA、体表抗原SA、分泌排泄抗原-ES)。SDS-PAGE结果表明,BA、HA、SA分子量在12~100kD之间,ES在12.3~26kD之间,蛋白质染色BA、HA、SA、ES分别显示25、24、18、9条带;IEF分析肝片吸虫分别得34、33、28、22条带,主要由酸性蛋白质组成,主要谱带在pI4.20~6.55内;双向电泳分析BA、ES多肽斑点为69、30个  相似文献   

7.
PCR扩增猪ESR基因一个外显子DNA片段的研究   总被引:4,自引:0,他引:4  
根据文献资料,参考人,鸡,鼠等动物的DNA序列,结合本研究的需要,我们设计了一对引物,其上游正向引物中P1为5′-GATACGAAAAGACCGCCGAG-3′(对应于鼠ESR基因第4外显子1010-1029)下游反向引物P2为5′-GCACTCTCTTTGCCCAGTTG-3′(对应于鼠ESR的基因第4外显子1322-1341)利用该引物对,按照首轮反应94℃5分钟,58℃1分钟,72℃1分钟,  相似文献   

8.
促卵泡素3号(LRH-A3)、人绒毛膜促性腺激素(hCG)和孕马血清促性腺激素(PMSG)等3种外源性激素均可增加体外培养条件下牦牛垂体组织分泌LH和FSH的能力。培养液中LH和FSH含量与加入的LRH-A3量呈正相关,与加入的PMSG和hCG的量无显著关系  相似文献   

9.
用赭曲霉素A(OA)-BSA合成抗原免疫Balb/c小鼠,取免疫鼠脾细胞与sp2/0细胞融合,通过克隆和ELISA法筛选,建立三株泌抗OA单克隆抗体杂交瘤细胞株(3C12,1D12和2G7)。用间接ELISA法测定,细胞上清液抗体效价为128^×(3C12)和64^×(1D12和2G7)腹水抗体效价为10^-7(3C12,1D12)和10^-6(2G7)。三株单抗(McAb)均属IgG类,分泌抗体  相似文献   

10.
用肝片吸虫排泄分泌抗原(ESAg)的酶联免疫吸附试验(ELISA)检测人工感染肝 吸虫的山羊血清中特异性IgG抗体动态变化。ESAg用量为13.8ug/孔,抗体稀释1 000倍,二抗1:2000稀释(3.5ug/孔),HRP标记的葡萄球菌A蛋白(SPA*)工作浓度为1:40。检测结果表明,2组实验山羊(第1组每只羊口服200 个 囊蚴,第2组每只羊口服500个囊蚴)在感染后第3周血清中的持异IgG即开始升高最  相似文献   

11.
Synchronization of development between the embryo and uterus is required for successful pregnancy establishment.Transfer of early embryos requires synchrony with the recipient uterus of 2 days or less in sheep,because asynchrony of 3 days or more results in failure of pregnancy recognition signaling for maintenance of corpus luteum (CL) and progesterone (P4) production and/or uterine support of the embryo.The objective was to determine if P4 treatment of recipient ewes would obviate the need for pregnancy recognition signaling and maintain a uterine environment conducive to embryo survival after asynchronous transfer,thereby establishing a universal recipient.Embryos (morulae/blastocysts) were recovered on day 6 from super-ovulated donor ewes.Recipient ewes received 25 mg P4 daily from day 6 post-estrus until 60 days after embryo transfer.Embryos were transferred into recipients on day 6,9,12,18,or 30 post-estrus.The pregnancy rate on day 22 post-transfer was 60% for synchronous transfers to day 6 ewes,44% and 22% for asynchronous transfers to day 9 and 12 ewes,and 0% for asynchronous transfers to day 18 and 30 ewes.On day 39 post-transfer,pregnancy rates remained 60% for day 6 ewes,33% for day 9 ewes,and 0% for day 12,18,and 30 ewes.The P4 treatment did extend the window of uterine receptivity to early embryos in ewes by one day,but did not create a universal recipient.Available results support the idea that a window of uterine receptivity to the conceptus exists in sheep that is independent of pregnancy recognition signaling.  相似文献   

12.
为探讨促孕散治疗持久黄体的作用机理,本研究通过直肠检查结合B超直肠检查对持久黄体奶牛做出诊断后口服促孕散,应用B超每3 d对黄体直径、卵泡数量、卵巢长度、卵巢宽度、子宫角纵径和子宫颈纵径进行测量,并统计1次,与用药前进行对比。30头持久黄体奶牛口服促孕散后,治疗有效头数25头,有效率为83.3%。停药后第1天卵巢出现小卵泡和中等卵泡,停药后分别在第10和19天大卵泡数量最多,部分奶牛出现发情并排卵,与用药前卵巢相比,结果发现左、右侧卵巢长在停药后均降低,但差异不显著(P>0.05);左、右卵巢宽在停药后均降低,但差异不显著(P>0.05);子宫颈纵径和子宫角纵径均在停药后升高,但差异不显著(P>0.05)。结果表明,B超是诊断奶牛持久黄体的有效手段,中药促孕散对持久黄体奶牛卵巢和子宫形态等指标具有一定的影响,可以促进黄体的溶解。  相似文献   

13.
In 1977 Bazer and Thatcher proposed that maternal recognition of pregnancy in the pig involves the secretion of PGF(2alpha) towards the uterine lumen (exocrine) rather than towards the uterine venous drainage (endocrine) as occurs in the non-pregnant pig during the mid to late stages of the estrous cycle. The retrograde transfer of PGF(2alpha) from the venous blood and uterine lymph into the uterus and the ability of the uterine vein and artery wall to accumulate PGF(2alpha) could constitute a part of putative mechanism of corpus luteum protection during early pregnancy. A luteotropic/anti-luteolytic effect of PGE(2) in the pig also has been frequently demonstrated and it seems that the most effective agent in changing PGE(2):PGF(2alpha) secretion is estradiol. The role for oxytocin during luteolysis and early pregnancy is controversial. It appears, however, that the main function of this hormone is autocrine and/or paracrine stimulation of PGF(2alpha) secretion. Pig trophoblastic interferons, unlike those of ruminants, do not themselves exert an anti-luteolytic effect in pigs. It is likely, that cytokines and angiogenic growth factors are involved in the initiation of luteolysis and/or maintenance of corpora lutea (CL).A discovery of functional LH receptors in porcine endometrium opened a new possibility for this hormone in luteolysis and perhaps in recognition of pregnancy in pigs. The endogenous LH pulses can provoke prostaglandin secretion from endometrium in pigs. On the other hand prolongation of up-regulation of LH receptors in endometrium of early pregnant gilts can additionally increase angiogenic factor production before the process of implantation is completed. Finally new integrated concepts of luteolysis and inhibition of luteolysis in pigs based on selectively reviewed information are presented.  相似文献   

14.
The effect of homogenized embryos on IUD-induced luteal regression in the ewe was studied. Plastic spiral IUDs were inserted in an uterine horn adjacent to an ovary containing a corpus luteum during various times of the estrous cycle. The ewes were sacrificed on Day 6 for examination of the corpus luteum. Estrous Cycle Day 3 was the last day in which the IUD insertion induced luteal regression. Corpus luteum was maintained in ewes when the IUD was inserted on Day 4, 5, or 6. Corpus luteum regression was not prevented when homogenized 14- or 15-day-old embryos were injected into the uterus at the time of IUD insertion. Ewes which received an IUD, a cannula, and an infusion of saline or homogenized embryos on Days 3, 4, and 5 maintained corpora lutea in 4 of 7 embryo-infused ewes and in none of 7 saline-infused ewes (p less than .05). It was determined that the IUD must remain in the uterine horn for longer than 1 day to induce corpus luteum regression and that embryos can sometimes prevent the IUD-induced regression.  相似文献   

15.
本研究旨在探明水牛唾液生殖激素、唾液结晶和卵泡发育变化的规律。应用ELISA试剂盒测定水牛唾液和血清中的生殖激素,制作唾液结晶,并分析生殖激素和唾液结晶与卵泡变化的关系。运用唾液结晶法、直肠检查法、唾液结晶+B超法、直肠检查+B超法4种方法在生产中进行验证。结果表明,水牛发情当天唾液雌激素(E2)的水平为233.51 pg/mL,达到一个峰值,随后开始缓慢降低。水牛唾液中孕激素(P4)水平在发情前2 d达到16.17 ng/mL,发情当天降到8.91 ng/mL。促卵泡素(FSH)在水牛发情前2 d逐渐下降,从37.91 ng/mL降到34.64 ng/mL,但在发情2 d后逐渐升高,达到61.20 ng/mL。促黄体素(LH)在发情前1 d逐渐下降,从5.15 ng/mL降至发情当天4.12 ng/mL,但在发情1 d后升至5.77 ng/mL。B超监测卵巢卵泡从发情前2 d迅速生长,到发情当天达到20 mm大小,直到破裂排卵卵泡的大小变化不显著。排卵后形成黄体,黄体期一直维持到下次发情前4 d左右,期间在发情后14~17 d的黄体最大,与水牛唾液中LH分泌峰一致。唾液结晶+B超鉴定方法判定的发情率最高(86.67%),与其他几种方法比较差异显著(P<0.05);妊娠诊断结果也表明,唾液结晶+B超鉴定方法判定的妊娠率(61.53%)显著高于直肠检查法、唾液结晶法和直肠检查+B超法3种方法鉴定的结果(P<0.05)。综上所述,卵巢上卵泡的发育与唾液中生殖激素的浓度显著相关,发情当天唾液结晶呈现典型的树枝状,唾液结晶+B超法的发情鉴定准确率最高。  相似文献   

16.
Maternal recognition of pregnancy refers to the requirement for the conceptus (embryo and its associated extra-embryonic membranes) to produce a hormone that acts on the uterus and/or corpus luteum (CL) to ensure maintenance of a functional CL for production of progesterone; the hormone required for pregnancy in most mammals. The pregnancy recognition signal in primates is chorionic gonadotrophin which acts directly on the CL via luteinizing hormone receptors to ensure maintenance of functional CL during pregnancy. In ruminants, interferon tau (IFNT) is the pregnancy recognition signal. IFNT is secreted during the peri-implantation period of pregnancy and acts on uterine epithelia to silence expression of estrogen receptor alpha and oxytocin receptor which abrogates the oxytocin-dependent release of luteolytic pulses of prostaglandin F2-alpha (PGF) by uterine epithelia; therefore, the CL continues to produce progesterone required for pregnancy. Pig conceptuses secrete interferon delta and interferon gamma during the peri-implantation period of pregnancy, but there is no evidence that they are involved in pregnancy recognition signaling. Rather, pig conceptuses secrete abundant amounts of estrogens between Days 11 to 15 of pregnancy required for maternal recognition of pregnancy. Estrogen, likely in concert with prolactin, prevents secretion of PGF into the uterine venous drainage (endocrine secretion), but maintains secretion of PGF into the uterine lumen (exocrine secretion) where it is metabolized to a form that is not luteolytic. Since PGF is sequestered within the uterine lumen and unavailable to induce luteolysis, functional CL are maintained for production of progesterone. In addition to effects of chorionic gonadotrophin, IFNT and estrogens to signal pregnancy recognition, these hormones act on uterine epithelia to enhance expression of genes critical for growth and development of the conceptus.  相似文献   

17.
18.
黄体释放的黄体酮对母畜发情周期、启动发情行为和维持妊娠起着至关重要的作用,但目前国内外对黄体是否影响胚胎移植效果有着不同报道。本文以探究黄体对牛胚胎移植效果的影响为主线,查阅大量文献,归纳黄体的检查和级别判断方法,分别分析黄体大小、黄体位置、黄体质量和黄体生产溶解过程激素水平与胚胎移植受胎率的关系,为提高胚胎移植效果提供科学的参考依据。生产上常通过B超扫描结合直肠检查法判断受体牛黄体发育和位置作为能否移植的依据;由各学者研究结果可见,不同直径黄体的胚胎移植受胎率差异不显著,为提高受体牛的利用,在对受体牛黄体大小选择时可适当放宽标准;胚胎移植到黄体同侧的受胎率比移植到黄体对侧的高,另外,可能因为操作者左右手操作习惯不同,发现左侧黄体移植受胎率比右侧的要高;较多学者认为质量好的胚胎选择黄体质量好的受体移植才能得到更高的受胎率;胚胎移植后能否成功受孕,还与黄体生成溶解过程中孕酮含量及孕酮与雌二醇的比例等有关。  相似文献   

19.
不同等级黄体与水牛胚胎移植受胎率关系的研究   总被引:1,自引:0,他引:1  
[目的]为了解胚胎移植过程中不同等级黄体与水牛胚胎移植受胎率之间的关系,为提高水牛受胎率提供依据.[方法]在水牛胚胎移植过程中,利用B超对受体卵巢进行扫描,确定黄体的有无并根据黄体大小对其分级.黄体直径在1.5 cm以上的定为A级,直径在0.8~1.5 cm之间的定为B级,直径在0.8 cm以下的定为C级.测定受体移植...  相似文献   

20.
莱芜黑山羊发情周期中FSH、LH、E2和P的分泌规律   总被引:6,自引:0,他引:6  
莱芜黑山羊在发情期和间情期,血浆内的卵泡刺激素(FSH)和黄体生成素(LH)均呈脉冲式分泌,雌激素(E2)和孕嗣(P)为波动式分泌。发情期FSH的脉冲周期较间情期长,两者之间显著差异(P〈0.05)。发情期LH的脉冲周期短于间情期,两者之间差异不显著(P〉0.05)。在整个发情周期中,FSH和LH均先后出现4个分泌峰,FSH和LH的第1个分泌峰分别出现在第7天和第5天,其余3个分泌峰均同时出现(分别为第10、15和20天)。E2和FSH、LH均在发情周期的第20天迭最高峰。P在间情期一直维持在一个较高水平。  相似文献   

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