Expression of Steroid Hormone Receptors in the Genital Structures of a True Hermaphrodite Pug Dog

Hermaphroditism is a rare and a not well‐understood disordered sexual development (DSD) in dogs. The objective of the study was to analyse the sex steroid hormone receptor (STHR) expression patterns in the internal genital structures, because the responsiveness of the different tissue types to the steroid hormones may have a key role in pathological alterations based on DSDs. Furthermore, the adhesion molecule β‐catenin was investigated by means of immunohistochemistry because of its important role in development, tissue integrity and disease. Molecular sexing was performed via PCR targeting DBX/DBY genes to identify the pug dog as a true XX hermaphrodite. The portions of uterine tissue revealed comparable expression patterns for STHRs as investigated in normal female reproductive tissue. In the male parts, β‐catenin showed strong expression in the Sertoli cells of the seminiferous tubules; this was in contrast to normal testicular tissue. Likewise, the layers of smooth muscle actin‐positive cells surrounding the seminiferous tubules were reduced in the hermaphrodite. The results of this study deepen the knowledge of tissue characteristics in a hermaphrodite dog and highlight the importance of early diagnosis because the STH responsiveness in maldeveloped reproductive tissue might lead to serious problems for the dog.     

Serum and Tissue Steroid Hormone Levels in Canine Mammary Tumours: Clinical and Prognostic Implications

Hormonal dependency of canine mammary tumours (CMT) has been studied over the last few decades. However, studies assessing the prognostic and predictive potential of serum and/or tissue steroid hormone levels are still scarce in CMT. To the best of our knowledge, this is the first report relating serum and tissue levels of steroid hormones and prognosis in dogs. Serum and tumour tissue from 45 female dogs with spontaneous CMT were included in the study. Moreover, serum and normal mammary tissue from 13 healthy female dogs were also included as controls. Steroid hormones were determined by competitive enzyme immunoassay. Overall, levels of steroid hormones in serum and tissue homogenates were significantly different between malignant and benign mammary tumours (p < 0.01), except for progesterone (P4) serum levels that revealed no statistical differences between groups. In malignant tumours, oestrone sulphate (SO4E1), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T) and P4 elevated tissue concentrations were significantly associated with tumour relapse and/or distant metastasis during follow‐up. A significant association was found between elevated tissue SO4E1 (p = 0.003), 17β‐oestradiol (E2) (p = 0.036), DHEA (p = 0.022), A4 (p = 0.001) and P4 (p = 0.013) concentrations and shorter disease‐free survival and overall survival in female dogs with malignant mammary tumours. The high levels of tissue steroids found in cases of poor prognosis open the possibility of additional new therapeutic approaches. Future clinical trials will be needed to clarify the usefulness of targeting steroid hormones in the treatment of this neoplastic disease.     

Presence and Distribution of Urocortin and Corticotrophin‐Releasing Hormone Receptors in the Bovine Thyroid Gland

Urocortin (UCN), a 40 amino acid peptide, is a corticotrophin‐releasing hormone (CRH)‐related peptide. The biological actions of CRH family peptides are mediated via two types of G‐protein‐coupled receptors, CRH type 1 (CRHR1) and CRH type 2 (CRHR2). The aim of this study was to investigate the expression of UCN, CRHR1 and CRHR2 by immunoprecipitation, Western blot, immunohistochemistry and RT‐PCR in the bovine thyroid gland. Immunoprecipitation and Western blot analysis showed that tissue extracts reacted with the anti‐UCN, anti‐CRHR1 and anti‐CRHR2 antibodies. RT‐PCR experiments demonstrated that mRNAs of UCN, CRHR1 and CRHR2 were expressed. UCN immunoreactivity (IR) and CRHR2–IR were found in the thyroid follicular and parafollicular cells and CRHR1‐IR in the smooth muscle of the blood vessels. These results suggest that a regulatory system exists in the bovine thyroid gland based on UCN, CRHR1 and CRHR2 and that UCN plays a role in the regulation of thyroid physiological functions through an autocrine/paracrine mechanism.     

Profile of Steroid Receptors and Increased Aromatase Immunoexpression in Canine Inflammatory Mammary Cancer as a Potential Therapeutic Target

Canine inflammatory mammary cancer (IMC) has been proposed as a model for the study of human inflammatory breast cancer (IBC). The aims of this study were to compare the immunohistochemical expression of aromatase (Arom) and several hormone receptors [estrogen receptor α (ERα), estrogen receptor β (ERβ), progesterone receptor (PR) and androgen receptor (AR)], in 21 IMC cases vs 19 non‐IMC; and to study the possible effect of letrozole on canine IMC and human inflammatory breast cancer (IBC) in vitro using IPC‐366 and SUM‐149 cell lines. Significant elevations of the means of Arom Total Score (TS), ERβ TS and PR TS were found in the IMC group (p = 0.025, p = 0.038 and p = 0.037, respectively). Secondary IMC tumours expressed higher levels of Arom than primary IMC (p = 0.029). Non‐IMC PR‐ tumours contained higher levels of Arom than non‐IMC PR+ tumours (p = 0.007). After the addition of letrozole, the number of IMC and IBC cells dropped drastically. The overexpression of Arom found and the results obtained in vitro further support canine IMC as a model for the study of IBC and future approaches to the treatment of dogs with mammary cancer, and especially IMC, using Arom inhibitors.     

Canine Tumour Suppressor Gene p53 - Mutation in a Case of Adenoma of Circumanal Glands

Highly conserved regions of the tumour suppressor gene p53, including the typical human tumour hot spots (codons 175, 245, 248, 249, 273 and 282), were investigated in various canine neoplasms. A mutation CGG TGG (arginine tryptophan) was detected in codon 249 in an adenoma of the circumanal gland.     

Histological Study of Canine Mammary Gland During the Oestrous Cycle

The development of the mammary gland is a puzzling phenomenon and the research on this field has been focussed mostly on the carcinogenesis, with a less goal‐oriented concern in basic histology. In order to determine the histological features of normal mammary gland in the different oestrous phases we used 39 non‐pregnant female dogs of various breeds and ages. The animals were grouped in: pre‐pubertal, pro‐oestrous, oestrous, early and late dioestrous, early and late anoestrous phases. Major changes of the canine mammary histology throughout the oestrous cycle were identified in this study. A rudimentary gland with few ducts in the base of the nipple was observed in pre‐pubertal female individuals and pubertal pro‐oestrous female ones. In the oestrus, small inactive lobules associated with ductal branching and inconspicuous regressive changes were observed, while in early dioestrus, a ductal arborization was present. In late dioestrus, a complete lobuloalveolar differentiation and secretory capacity was achieved. The regressive histological features were abundant on early anoestrus, and markedly generalized on late anoestrus. The regressive process was longer in the more caudal gland pairs. This work provides baseline knowledge of canine mammary gland that may be relevant for interspecies comparative purposes and for pathologists dealing with mammary gland tumours.     

Growth Hormone Receptors in the Porcine Testis During Prepuberty

Supplementation of exogenous growth hormone (GH) during prepuberty advances onset of spermatogenesis in boars, but the mechanism of action is unknown. The present study is an investigation of the presence and characteristics of testicular growth hormone receptors (GHR). A total of 36 boars were castrated, three boars every 10 days, between the ages of 10 and 120 days. Testicular membrane preparations of 10, 20, 30, 50, 70, 100 and 120‐day‐old boars were used to determine ¹²⁵I‐bGH binding and Scatchard analysis. Liver from a 60‐kg barrow was used for comparison. Specific ¹²⁵I‐bGH binding to testicular membrane preparations occurred in all age groups with the exception of 20‐day‐old boars at levels of 30–40% of liver binding. At 30 days of age the unlabelled bGH at 1.1 ng/tube achieved half maximal inhibition (ID₅₀). Results of Scatchard analysis indicated a single class of binding sites. Binding affinity was 2.89 × 10⁹m with a binding capacity of 12 fmole/mg membrane protein. The results from this study suggest that GH may act directly on the cells of the prepubertal boar testis.     

家禽睾丸主要生殖激素受体的研究进展

睾丸的主要功能是制造精子,而睾丸中生殖激素与其受体的结合又影响着精子的发生和成熟,因此家禽睾丸中生殖激素受体的研究对提高其受精率具有十分重要的意义。本文主要阐述四类生殖激素受体——促卵泡激素受体、促黄体激素受体、雄激素受体和雌激素受体的研究进展,为提高家禽繁殖力,推进家禽养殖业发展提供一定的参考。     

Histochemical Detection of Glycoconjugates in the Canine Epididymis

A histochemical study using fluorescein isothiocyanate (FITC)-labelled lectins to identify glycoconjugates present in the efferent ductules and the three segments of the ductus epididymis (initial, middle and terminal segment) of dogs was carried out. The lectins used were: mannose-binding lectins (Con A, LCA and PSA), galactose-binding lectins (PNA, RCA), N -acetylgalactosamine-binding lectins (DBA, SBA, SJA and GSL I), N -acetylglucosamine-binding lectins (WGA and WGAs), fucose-binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E and PHA L). The lectin-binding pattern in the canine epididymis presents similarities and differences to those observed in other mammalian species. The ductuli efferentes distinctly stained with most of the lectins used, whereas in the ductus epididymis a segment specific staining pattern was observed. Whereas principal cells of the ductus epididymis stained clearly with several FITC-labelled lectins (WGA, UEA and PHA-L), basal cells showed only a significant binding of Con A.     

犬乳腺肿瘤的病理组织学观察及p63蛋白的表达

通过对35例犬乳腺肿瘤的临床病史调查、光学显微镜观察和免疫组化染色,探讨了犬乳腺肿瘤的临床病理形态学特点、诊断和鉴别诊断.结果显示34例发生在5岁以上,6岁～10岁为发病高峰期;51.4％的病例为恶性肿瘤,其中44.4％的乳腺癌发生转移.免疫组化染色结果显示,在正常乳腺和良性肿瘤组织中,p63蛋白100％表达,并且仅仅...     

犬场蚊子与犬细小病毒

２０００年４月，应用套式ＰＣＲ技术从犬场蚊子的血液样品检测出犬细小病毒。将分离到的犬细小病毒ＶＰ２－Ｙ３４基因片段克隆到ＰＭＤ１８－Ｔ载全，其病毒基因组ＣＰＶ－ＰＶ２－Ｙ３４序列测定结果显示与作者在ＧＥＮＥＢＡＮＫ发表的肺病变犬细小病毒ＣＰＶ－ＨＮ－１株有９９％同源性。     

促黄体素受体在母畜生殖道的分布与表达

在多种动物包括灵长类动物、猪、牛、火鸡和人,子宫内都发现有促黄体素(LH)或人绒毛膜促性腺激素(hCG)及其mRNA的结合位点.牛、猪在整个发情周期子宫内都存在LH受体(LHR)蛋白及其mRNA,并在黄体期表达最高.LH通过激活腺苷酸环化酶和磷脂酶C信号途径,增加前列腺素合成酶(PGSH),也称环加氧酶(COX)的活性,从而促进前列腺素(PG)的合成.牛、猪、羊在发情周期的15 d左右PGF2α升高,这与子宫内LHR的活化有关.在不同种动物的输卵管、子宫内膜、子宫肌层和子宫颈,LHR的释放呈现出动态特征,表明LH在发情周期和胚胎附植的分子自分泌-旁分泌调节中可能具有重要的作用.     

乳制品中类固醇激素类化合物检测研究进展

类固醇激素又称甾体激素,是一种亲脂性的小分子物质,常作为促生长剂被违法添加,用于畜牧业和养殖业。乳制品中残留激素可通过食物链进入人体并产生一系列健康危害。乳制品中类固醇激素的残留在国内外已成为兽药残留问题关注的重点。我国在乳制品中的类固醇激素含量限制方面已颁布相关法规。本文综述类固醇激素的结构、分类及功能,以及乳制品中类固醇激素限量标准、检测方法、含量及健康风险研究进展,旨在为乳制品中的类固醇激素研究提供借鉴和参考。     

Thyroid Hormone Concentrations In Critically III Canine Intensive Care Patients

Alterations in thyroid indices in critically ill dogs were studied retrospectively to determine the incidence of the euthyroid sick syndrome and to assess its prognostic relevance to survival. Sixty-seven dogs were classified as euthyroid bases on a TSH stimulation test. Forty-one of 67 (61%) of the euthyroid dogs had low baseline serum T₄ concentrations (<1.5 ug/dl) and 38 of 67 (56%) of the euthyroid dogs had low baseline serum T₃ concentrations (<75 ng/dl). There was no significant difference between either the baseline or the post-stimulation serum T₄ concentrations when values of dogs that survived were compared with those that died. There was, however, a significant difference (P<0.05) between both the baseline and the post-stimulation values for T3 when survivors (100.3± 81 ng/dl, 143± 66 ng/dl) and non-survivors (64.2 ± 17.1 ng/dl, 96.6 ± 38.3 ng/dl) were compared. The euthyroid sick syndrome occurs commonly in critically ill dogs. Further, the extent of depression of serum concentrations of T₃ may be correlated to mortality.     

Heat Shock Protein 70 and Sex Steroid Receptors in the Follicular Structures of Induced Ovarian Cysts

The purpose of this study was to estimate the expression and relative amounts of estrogen (ER) and progesterone receptors (PR) and their isoforms as well as heat shock protein 70 (HSP70) in ovaries of rats with induced cystic ovarian disease (COD). Primary, secondary, tertiary, atretic and cystic follicles were evaluated by immunohistochemistry and total ovarian proteins were analyzed by Western blot. In the granulosa layer, growing and cystic follicles in the treated group have a higher expression of ERα than growing follicles of control individuals. In the theca interna layer, tertiary follicles presented a significantly higher expression of ERα in the treated group. An increase in total ERα protein was detected in the treated group. Granulosa cells of all growing, atretic and cystic follicles show a lower expression of ERβ in animals with COD, and the total protein expression of ERβ was lower in this group. The expression of PR was lower in the granulosa cell layer of tertiary and cystic follicles in treated animals, and theca interna layer had less intense immunostaining in this group. Although there were no differences in the expression of PR-B by Western blotting, the expression of PR-A was higher and the expression of PR-C was smaller in the treated group. An intense HSP70 immunostaining was observed in the cells of cystic follicles. By Western blotting, higher protein expression of HSP70 was detected in the ovarian samples of the control group than those of the treated ones. Ovaries of animals with COD exhibited an altered steroid receptor expression and subtype balance as compared with control animals, and an increase in HSP70 immunoexpression.     

犬巴贝斯虫病检测研究

犬巴贝斯虫病是经蜱传播的血液寄生虫病,其临床症状是高热、贫血、血红蛋白尿和脾肿大。通过血涂片检验,血常规检验并结合临床特征,对西安市103只宠物犬进行了巴贝斯虫的感染情况检测。结果显示,犬血液中巴贝斯虫的感染数为16例,占总数的15.53%,其中5月份的感染犬最多,为13例,占感染数的81.25%(13/16)。     

核酸水平检测犬副流感病毒方法的建立

根据GenBank中与犬副流感(CPIV)同源性较近的SV5病毒N基因保守序列,利用DNAStar软件设计了一对特异性引物,能扩增265bp大小的片段,并以此建立了检测CPIV的RT-PCR方法,实验证明该对引物特异扩增CPIV;不扩增犬瘟热病毒、犬细小病毒、犬腺病毒和狂犬病病毒犬的四种病原的核酸。检测临床病料20份,其中2分为CPIV阳性。此法敏感性较高。是检测犬急性传染性呼吸道疾病(CIRD)中CPIV的有效的方法。     

Detection and Sequence Analysis of Canine Circovirus in Chongqing

The aim of this study was to investigate the prevalence of canine circovirus (CCV) in Chongqing in recent years,and to explore the characteristics of the field strains in Chongqing.In this study,100 dog serum samples collected in Chongqing in 2017 were tested for CCV nucleic acid by PCR,and the obtained CCV positive samples were amplified and sequenced with the full length genome,and the Chongqing CCV strain isolated was analyzed by MegAlign,Mega 6.0 and RDP4 software.Four positive sample were found,with a positive rate of 4%,and three full length CCV genomes (CQ76,CQ79,and CQ82) were obtained,with a total length of 2 062 nt.Compared with the previously reported genome of CCV with a length of 2 063 nt,one base was missing,which was located downstream of the stem ring structure in the 5'-intergenic region.The length of the 5'-intergenic region and 3'-intergenic region between the two ORFs of the three strains were 134 (1 929-2 062 nt) and 203 nt (913-1 115 nt),respectively.The homology of CQ76,CQ79,and CQ82 ranged from 99.8% to 100%,among which,CQ76 and CQ82 only had synonymous mutations at the 2 019 locus.The homology of genome sequences of the three Chongqing strains and other strains was 82.7% to 97.1%,among which,the degree of variation of ORF2 was greater than that of ORF1.Based on virus ORF2 sequences and genome-wide respectively build Neighbor-Joining in the evolutionary tree,CQ76,CQ79 and CQ82 all belonged to the same subgroup,which was close to the Chinese strain 204,which belonged to genotype Ⅱ,and the homology was 96.5% to 96.7%.RDP4 recombinant analysis indicated that the genomes of CQ76,CQ79 and CQ82 strains were all the recombinant sequences of 204 and 388 strains from Guangxi,and the recombinant region was located in ORF2.Above results enriched the epidemiological and genetic evolution information of CCV and provided further basic data for prevention and control research.     

重庆地区犬圆环病毒检测及序列分析

本试验旨在调查近年来新发的犬圆环病毒（canine circovirus,CCV）在重庆地区的流行情况,探索重庆流行毒株的特性。本研究利用PCR方法对2017年采集于重庆地区的100份犬血清样品进行CCV核酸检测,对所得CCV阳性样品进行全基因组扩增与测序,并利用MegAlign、Mega 6.0和RDP4等软件对分离到的重庆CCV毒株进行分析。结果显示,重庆地区100份犬血清中共有4份为CCV阳性,阳性率为4%,从4份阳性样品中共获得3株不同的CCV基因组（CQ76、CQ79和CQ82）,全长基因组序列均为2 062 nt,与此前报道长度为2 063 nt的CCV基因组相比缺少了一个碱基,该碱基位于5'-基因间隔区内茎环结构的下游。3个毒株的两个ORF之间5'末端间隔区和3'末端间隔区长度分别为134（1 929-2 062 nt）和203 nt（913-1 115 nt）。CQ76、CQ79和CQ82的同源性在99.8%~100%之间,其中CQ76与CQ82仅在第2 019位点存在同义突变;所获得的3个重庆毒株与国内外报道的其他CCV基因组序列同源性为82.7%~97.1%,其中,ORF2的变异程度大于ORF1。基于病毒ORF2序列和全基因组分别构建NJ进化树中,CQ76、CQ79和CQ82均属于同一亚群,与属于基因Ⅱ型的中国毒株204株遗传距离较近,同源性为96.5%~96.7%。RDP4重组分析发现,CQ76、CQ79和CQ82毒株基因组均为广西毒株204株和388株的重组序列,重组区域坐落于ORF2。本研究为丰富CCV流行病学和遗传进化信息,以及进一步防控研究提供基础数据。