Cortisol Concentrations in Well‐Regulated Dogs with Hyperadrenocorticism Treated with Trilostane

Background

There are no clear treatment guidelines for dogs with clinically well‐regulated hyperadrenocorticism in which serum cortisol concentrations before and after an ACTH stimulation test performed 3–6 hours after trilostane administration are < 2.0 μg/dL.

Objective

To determine if serum cortisol concentrations measured before (Pre1) and after (Post1) ACTH stimulation at 3–6 hours after trilostane administration are significantly lower than cortisol concentrations measured before (Pre2) and after (Post2) ACTH stimulation 9–12 hours after trilostane administration, in a specific population of dogs with clinically well‐regulated hyperadrenocorticism and Pre1 and Post1 <2 μg/dL.

Animals

Thirteen client‐owned dogs with clinically well‐regulated hyperadrenocorticism and Pre1 and Post1 serum cortisol concentrations <2.0 μg/dL 3–6 hours after trilostane administration.

Methods

Prospective study. Dogs had a second ACTH stimulation test performed 9–12 hours after trilostane administration, on the same day of the first ACTH stimulation test. Cortisol concentrations before and after ACTH stimulation were compared using a paired t‐test.

Results

Cortisol concentrations before (1.4 ± 0.3 μg/dL) and after the first stimulation (1.5 ± 0.3 μg/dL, mean ± SD) were significantly lower than cortisol concentration before the second stimulation (3.3 ± 1.6 μg/dL, P = .0012 each). Cortisol concentration before the first stimulation was also significantly lower than cortisol concentration after the second stimulation (5.3 ± 2.4 μg/dL, P = .0001).

Conclusions and clinical importance

In dogs with clinically well‐regulated, trilostane‐treated, hyperadrenocorticism, and cortisol concentrations <2 μg/dL before and after the first stimulation, a second ACTH stimulation test performed 9–12 hours after treatment can result in higher cortisol concentrations that could support continued trilostane treatment.     

白羽瘤头鸭不同生长阶段的甲状腺活动

本文报道了用放射免疫分析法测定５７例不同日龄的白羽瘤头鸭（Ｃａｉｒｉｎａｍｏｓｈａｔａ）外用血清中Ｔ４的含量和用组织学方法检查甲状腺滤泡的发育作为甲状腺活动的指标，研究不同日龄的生长速度与甲状腺活动的关系。５７例白羽瘤头鸭的血清Ｔ４含量平均为１．３６±０．３７μｇ／１００ｍｌ，其中１—４０日龄甲状腺的分泌活动除２０日龄外，其他的均随着日龄增长而升高，分泌的高峰出现在４０日龄，以后随日龄增长而下降。４０日龄组血清Ｔ４的含量与１、１０、２０、６０、和９０日龄组比较，差异极显著（Ｐ＜０．０１），血清Ｔ４的波动与相对生长率之间有统计学强的正相关，差异极显著（Ｐ＜０．０１）。组织切片检查，刚出壳的雏鸭甲状腺滤泡小。１０—４０日龄甲状腺滤泡随日龄的增长而增大，滤泡腔被甲状腺球蛋白所组成的蛋白质凝胶的均质液所充满、着色深而均匀。６０—９０日龄鸭甲状腺滤泡虽然增大，但其内部凝胶的均质液充满不完全或空虚，着色浅而不均匀。各日龄组甲状腺滤泡的变化和甲状腺分泌的含量变化与Ｔ４的波动相一致。     

甲状腺素对猪小肠上皮细胞miR-let-7a基因表达及细胞增殖的影响

miR-let-7a在动物细胞的分化、增殖与凋亡等方面发挥越来越重要的作用。甲状腺激素(TH)作用非常广泛,机体的每个细胞几乎都是TH作用的靶细胞,其可以促进组织分化、生长和成熟。本实验用甲状腺素(T4)浓度分别为(0、0.02、0.03、0.05、0.075、0.1、0.2μmol/L)在体外培养猪的小肠上皮细胞。结果表明:T4处理组的细胞体积形态相对于空白对照组没有明显变化;当T4添加浓度为0.03μmol/L时,细胞的增殖率显著低于其他组(P0.05);当T4浓度为0~0.03μmol/L时,let-7a的表达随着添加剂量的增加而升高,浓度从0.03~0.2μmol/L变化时,let-7a的表达呈现降低趋势,浓度为0.03μmol/L时表达量极显著高于其他组(P0.01)。let-7a的表达量与细胞增殖呈负相关。     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

GnIH与INH表位多肽疫苗主动免疫对甘肃高山细毛羊生殖激素的影响

本试验通过合成促性腺激素类的抑制激素——抑制素(inhibit hormone,INH)与促性腺激素抑制激素(gonadotropin inhibiting hormone,GnIH)的INH表位多肽疫苗和GnIH表位多肽疫苗,主动免疫甘肃高山细毛羊并对促卵泡素(follicle stimulating hormone,FSH)和促黄体素(luteinizing hormone,LH)激素水平进行比较分析。试验选用15只处在非发情季节的3.5岁的甘肃高山细毛羊,注射INH表位多肽疫苗与GnIH表位多肽疫苗分别为表位多肽疫苗A组和表位多肽疫苗B组,对照组注射等量生理盐水。采血后进行免疫,每隔7 d免疫和采血,同时在发情后每隔15 min采集一次血液,分离血清,用ELISA试剂盒检测抗体水平及FSH和LH激素变化。结果表明,表位多肽疫苗A组和表位多肽疫苗B组在初次免疫后都产生了相应的抗体,且表位多肽疫苗B组较表位多肽疫苗A组产生的抗体浓度高,75 、90、105 min时,表位多肽疫苗B组相对表位多肽疫苗A组显著促进了FSH的分泌水平(P<0.05)。在45~105 min时,表位多肽疫苗B组相对表位多肽疫苗A组显著促进了LH的分泌水平(P<0.05),且在90到105 min时表位多肽疫苗A组和表位多肽疫苗B组血清中FSH和LH的水平均显著高于对照组(P<0.05)。本试验结果表明,INH表位多肽疫苗和GnIH表位多肽疫苗主动免疫促进了甘肃高山细毛羊的FSH、LH的分泌,GnIH表位多肽疫苗在甘肃高山细毛羊上具有更好的免疫作用,本研究为该表位多肽疫苗在绵羊上的运用奠定了基础。     

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.     

西藏牦牛和黑白花奶牛生长激素基因AluⅠ多态性的比较研究

利用聚合酶链式反应 (PCR)技术 ,测定了西藏牦牛和黑白花奶牛生长激素基因的AluⅠ RFLP (限制性内切酶片段长度多态性 ) ,并比较了2牛群的基因频率。在西藏牦牛和黑白花奶牛中 ,等位基因A的频率分别为 0 935和 0 90 5 ,等位基因B的频率分别为 0 0 65和 0 0 65 ,卡方检验差异不显著 (P >0 0 5)。     

蓝塘仔猪IGF-1水平与组织IGF-1、GHR基因的表达

32头不同日龄(出生、3、21、35d)蓝塘仔猪,由前腔静脉采血后剖杀,取肝脏、背最长肌样品。用RIA法测血液、组织中IGF 1浓度,用放射受体法(RBA)检测肝脏、肌肉组织中GHR结合活性,用实时荧光定量PCR法检测IGF 1、GHRmRNA的表达水平。结果表明:(1)血液中IGF 1在出生日显著高于其它时期(P<0 05)。肌肉组织中IGF 1含量高于肝脏组织,肌肉组织IGF 1含量在3、21、35日龄时都显著高于出生日(P<0 05)。(2)肝脏细胞膜GHR结合活性在出生日显著高于3、21日龄(P<0 05),肝脏细胞膜GHR结合活性高于肌肉组织。(3)肝脏组织IGF 1、GHRmRNA的表达量均显著高于肌肉组织(P<0 05)。肝脏IGF 1mRNA的表达在出生日、21日龄时显著高于3、35日龄(P<0 05),GHRmRNA的表达在出生日显著高于其它日龄(P<0 05)。肌肉IGF 1、GHRmRNA的表达在出生日均显著高于其它日龄(P<0 05)。     

表达猪生长激素基因的重组腺病毒的构建及对猪生长的促进效果观察

利用复制缺陷型重组腺病毒作为基因表达载体,将猪生长激素（pGH）基因直接转导到猪体内细胞,研究猪生长激素基因在猪体内的促生长作用.用同源重组的方法,构建含pGH基因的重组腺病毒（Ad-pGH）.将其感染293细胞和PK-15细胞,均可检测到pGH.用1 mL此重组腺病毒（1010TCID50）一次性肌肉注射60日龄的杂交长白猪.结果表明,试验组的猪平均增重比对照组的猪增加37%（注射后第4周）和19%（注射后第6周）;饲料报酬提高28%（注射后第4周）和18%（注射后第6周）.试验结果证明,由重组腺病毒表达的pGH具有生物学活性,在猪体内起到明显的促进生长的作用.重组腺病毒介导的pGH基因在猪体内的表达可维持约4周.     

内源激素变化及外源生长素对紫花苜蓿种子萌发过程的影响

为了探索紫花苜蓿萌发过程内源激素的变化趋势及生长素对其萌发的影响,本研究以‘新牧4号’紫花苜蓿（Medicago sativa L.‘Xinmu No.4’）为试验材料,采用酶联免疫吸附法（ELISA）测定未萌发种子（M1）、露白种子（M2）、肧根长度1.0~1.2 cm种子（M3）、苗期种子（M4）这四个萌发时期种子中内源激素的含量;并研究不同浓度IAA浸种处理对种子萌发及子叶中内源激素的影响。结果表明：种子萌发过程中,赤霉素含量逐渐增多,脱落酸含量逐渐减少,但在M4时期含量略有回升;生长素含量先减少随后逐渐增多。不同浓度IAA浸种处理后,10 mg·L^-1 IAA可显著提高种子的萌发势、萌发率,促进幼苗的生长,与对照相比,子叶中脱落酸含量显著减少（P<0.05）,生长素、赤霉素含量显著增多（P<0.05）。综上所述,10 mg·L^-1 IAA浸种处理能促进苜蓿种子萌发,并促进子叶中生长素、赤霉素的合成。