双拷贝抑制素基因免疫对大鼠卵泡发育、产仔及生殖激素的影响

将90只大鼠随机分为5组,分别注射10μg.100μL-1 pcISI(T1)、50μg.100μL-1 pcISI(T2)、100μg.100μL-1(T3)pcISI、100μg空载体pcMV-S(V)和100μL生理盐水(S),以探讨在不使用免疫佐剂的情况下,不同剂量的双拷贝抑制素pcISI基因免疫对大鼠卵泡发育、产仔和生殖激素的影响。结果表明,加强免疫能提高抗体P/N值,加强免疫后各剂量组平均抗体P/N值大于2,T3组抗体P/N值显著高于T1和T2组(P<0.05)。T3组成熟卵泡数显著高于对照组(P<0.05),且抗体阳性鼠成熟卵泡数比阴性鼠多12.45个(P<0.05)。除了T1组外,其他2个剂量组产仔数和胎盘数均高于对照组(P<0.05)。抗体阳性鼠胎盘数比阴性鼠多5.09个(P<0.05),产仔数比阴性鼠多5.39个(P<0.05)。抑制素pcISI基因免疫大鼠后促卵泡素(FSH)、雌二醇(E2)和孕酮(P4)含量均高于对照组,T3组的FSH含量与对照组差异显著(P<0.05),T3组的E2含量与对照组差异极显著(P<0.01),各剂量组P4含量与对照组差异均不显著(P>0.05)。这些结果说明,在没有免疫佐剂的前提下,双拷贝抑制素pcISI基因免疫可产生抗体,促进FSH分泌和卵泡发育,增加产仔数。本试验条件下100μg.100μL-1是最佳免疫剂量。     

Granulosa cell tumour: An interesting case in a pregnant mare

This report describes the successful management of a pregnant 14‐year‐old seven‐eighths Thoroughbred mare with an ovarian granulosa cell tumour. The mare initially presented with unilateral ovarian enlargement whilst being managed for artificial insemination, demonstrating normal ovarian function with ovulation from the contralateral ovary leading to conception. The mare subsequently re‐presented with stallion‐like behaviour at 3.5 months gestation and ovarian suppression was evident. The mare maintained her pregnancy and delivered a live colt foal at term. Ovariectomy was performed 3 months post foaling and the mare regained cyclic activity 9 months post surgery. The mare then conceived and became pregnant once more. The diagnostic and therapeutic challenges during pregnancy are discussed.     

Establishment and Optimization of INH Epitope Peptide Vaccine for Detection of Antibody

In this study, an indirect ELISA method was established to detect inhibin hormone (INH) epitope peptide vaccine antibody, it would provide oretical reference for the determination of Fine-wool sheep after active immune body INH epitope peptide vaccine antibody. On the basis of the predecessors, using indirect ELISA method to determinate serum INH epitope peptide antibody levels of sheep, and through control different experimental conditions to look for the best experimental conditions. Through explorating the experimental conditions, finally, the testing experiment conditions were determined, which was blocked solution with skimmed milk powder, INH and GnIH synthetic peptides dilution degrees for 20 000 times, the optimum reaction time was 60 min, the best color action time was 15 min. In this experiment, a kind of method to detection antibody in the body after INH active immune sheep was built, it would provide a reference for future research.     

Downregulation of the expression of inhibin α subunit and betaglycan in porcine cystic follicles

Inhibins, as members of the transforming growth factor beta (TGF-β) superfamily, downregulate the synthesis and secretion of follicle-stimulating hormone (FSH) in an endocrine manner. The role of inhibin/betaglycan in the ovary regulation recently gained attention. To date, no data exist on the function of inhibin α subunit and betaglycan in cystic follicles. In this study, the expressions of inhibin α subunit and betaglycan in cystic follicles were investigated using immunohistochemistry, real-time PCR and Western blot analysis. Both inhibin α subunit and betaglycan immunoreactivities were mainly localized in the granulosa cells of follicles. Expression of inhibin α subunit and betaglycan was inferior in cystic follicles compared with that in normal large follicles. However, the result of enzyme-linked immunosorbent assay showed no significant difference in the decreasing in concentration of inhibin α subunit in cystic follicular fluid compared with the control (P>0.05). In this study, we explored the effects of FSH on betaglycan expression in granulosa cells in vitro. As expected, a significant increase in the expressions of betaglycan mRNA and protein in granulosa cells was observed in response to exogenous FSH (30 ng/ml) (P<0.05) compared with the control. Consequently, this study provides evidence that the expressions of inhibin α subunit and betaglycan are inferior in cystic follicles, and this may be caused by the decrease in FSH in the presence of a cystic follicle.     

抑制素免疫及其基因工程疫苗的构建

本文叙述了抑制素的特性、免疫原性及来源途径,并论述了其对提高家畜繁殖力的作用,同时提出了抑制素基因疫苗构建的方法及基因疫苗的优势。     

Induction of oestrus by administering Inhibin antiserum along with equine chorionic gonadotropin in anoestrous bitches

As dogs experience oestrus only once or twice a year, it is necessary to establish an effective method of oestrous induction for efficient breeding. In the present study, we evaluated inhibin antiserum (IAS) on oestrous induction in anoestrous females. Bitches were administered 0.5 ml/kg IAS or a mixture of 50 IU/kg equine chorionic gonadotropin (eCG) and 0.5 ml/kg IAS and 500 IU human chorionic gonadotropin (hCG) administered 7 days after the mixture injection. As a control, bitches received 50 IU/kg eCG, with 500 IU hCG administered 7 days after eCG injection. Blood-tinged vaginal discharge, vulvar swelling, plasma progesterone concentrations and ovarian follicular development were assessed from day 0 to day 14. IAS alone injection did not induce oestrus in bitches at the anoestrous stage. Conversely, vulvar swelling, blood-tinged vaginal discharge and an estimated luteinizing hormone (LH) surge appeared on days 3–7, days 3–6 and days 7–9 after the IAS+eCG mixture injection, respectively, in all five bitches at the anoestrous stage. The average number of developing and ovulated follicles in bitches administered IAS+eCG was 8.8 and 9.6 respectively. A single eCG injection followed by hCG induced oestrous signs, with an average of 8.3 developing follicles and 4.5 ovulated follicles. This study revealed that IAS alone did not induce oestrus, but when IAS was used in combination with eCG, it induced oestrus and promoted a considerable number of ovulations in anoestrous dogs.     

卵泡抑制素与GFP融合基因疫苗免疫对大鼠卵巢和生殖激素的影响

将50只大鼠随机分为5组,分别注射0.2（T1）、0.6（T2）、1g/L（T3）pEGISI,100μg空载体pEGFP-N1（C1）和100μL生理盐水（C2）,以探讨在没有使用免疫佐剂的情况下,抑制素与GFP融合基因疫苗pEGISI免疫对大鼠卵巢和生殖激素的影响。研究结果,加强免疫能提高抗体P/N值,T3组在加强免疫期与对照组差异显著（P〈0.05）;加强免疫第2周时T3组抗体阳性鼠比例达40%,与T1和T2组差异均显著（P〈0.05）。T3组卵巢大小和卵泡数均显著高于对照组（P〈0.05）,但抗体阳性鼠大卵泡数与阴性鼠差异不显著（P〉0.05）。各剂量组促卵泡素和雌二醇均高于对照组,且在加强免疫第2周时T3组与对照组差异显著（P〈0.05）;各剂量组孕酮含量与对照组差异均不显著（P〉0.05）。结果表明,在没有使用免疫佐剂的前提下,抑制素pEGISI基因免疫可产生抗体,促进卵巢发育和FSH分泌。本试验条件下100μg是最佳免疫剂量。     

抑制素基因免疫对南阳黄牛生殖激素的影响

选择40头南阳黄牛,用3.0 mg抑制素基因重组质粒(pCIS,免疫组,n=30)或同剂量的生理盐水(对照组,n=10)间隔21 d注射2次,分别在首次免疫当天、首次免疫后第10天和第21天、加强免疫后第10天和第45天收集血清,应用放射免疫法(RIA)测定处理后不同时期血清中激素水平.结果表明:在5个采血时间点内,加强免疫后第10天抗体阳性牛比率最高,为37.5%,与其他采血时间点有显著差异.免疫组促卵泡素(FSH)平均含量比对照组高,但差异不显著;17-β雌二醇(E2)和孕酮(P4)水平在加强免疫后2组有显著性差异.另外.抑制素抗体阳性牛的FSH、E2和P4比阴性牛高,但差异不显著.这些结果说明抑制素基因重组质粒可促进肉牛产生特异性抗体,且对FSH、E2和P4水平有影响.     

应用猪精液抑制素主动免疫法诱导黄牛孪生

 2 70头经产母黄牛分成 3组 ,每组 90头 ,分别在产后 13个月用 2mg(Th组 )或 1mg(Tl组 )精液抑制素和福氏完全佐剂进行主动免疫 ,34周后进行加强免疫。加强免疫所用抑制素剂量减半 ,佐剂为福氏不完全剂。另一组牛用不含抑制素生物活性的精液提取物和上述佐剂进行主动免疫和加强免疫 ,用作对照组 (C组 )。母牛发情时 ,间隔 8 12h用冷冻精液进行人工授精。在主动免疫和加强免疫后 812d采集颈静脉血 ,收集血清 ,分别用双扩散凝胶沉淀法和酶联免疫吸附测定法测定抗抑制素抗体的效价。根据记录完整的 2 4 7头牛资料 ,发现Th组 87头母牛中有 73头(83.9% )发情 ,其中 5 1头排单卵 ,19头排双卵。妊娠期末 ,15头产双胎 (孪生 2 6 .3% ,15 / 5 7)。而在T1组 (72头 )和C组 (88头 )中 ,分别只有 4 4头 (6 1.1% )和 5 5头 (6 2 .5 % )母牛发情 :其中排双卵的母牛分别为 4头和 0。妊娠期末 ,孪生率分别为 3.1% (1/ 32 )和 0 (0 / 34)。分析血清抗体水平 ,证明排双卵的母牛血清抗抑制素水平显著高于排单卵和不排卵的母牛 ,排卵数与抗抑制素抗体水平呈正相关 (r=0 .75 0 7,P <0 .0 1)。     

抑制素免疫对牛、羊繁殖力作用的研究进展

文章综述了天然提纯,化学合成及重组等不同种类抑制素抗原免疫对牛、羊繁殖力的影响.近年来抑制素疫苗作为多胎疫苗的角色已越来越为人们所重视,其免疫原性同抗原本身一些因素如纯度、免疫原性、异原性、免疫剂量有关.