A Screening for the Occurrence of Autoreactive Antibodies in Sera of Pregnant and Non-pregnant Bitches

Sera of healthy pregnant (group I, n = 11) and non-pregnant (group II, n = 11) bitches were screened for autoantibodies (AAb). In both groups, blood samples were drawn every fifth day between days 5 and 55 after mating. Serum was analysed via indirect immunofluorescence (IIF) with the Canine ANA HEp-2 Screening Kit. In all animals, anticytoplasmic AAb were detected. Utilizing primate-heart substrate slides AAb against contractile proteins of the cytoplasm could be observed. The predominating fluorescence pattern in pregnant animals resembled above all desmin, which was proven via Western blot. The sera were then pre-incubated with tropomyosin, actin, vimentin, vinculin and keratin solutions, and assessed on HEp-2 slides and on human and canine fibroblasts as well. The latter substrate was used to verify whether the detected Ab were in fact AAb. Utilizing tropomyosin, revealed elimination of the cytoplasmic fluorescences on all three substrates. It is therefore assumed, that in sera of healthy dogs, AAb against contractile structure proteins of the cytoplasm are present regularly. The majority of pregnant bitches presented with higher end-point titres (EPT), than to be found in non-pregnant dogs. AAb against desmin played the key role in those patterns. In addition, sera were screened for thyroid specific AAb, namely thyroglobulin, thyroid peroxidase (TPO), T3 and T4, and for AAb against insulin by ELISA or Western blot (TPO). Only in two of the pregnant bitches a weak positive reaction (1:100) for T3-AAb was detected.     

Changes in Blood Glucose,Plasma Non-esterified Fatty Acids and Insulin in Pregnant and Non-pregnant Goats

The blood glucose and the plasma non-esterified fatty acids (NEFA) and insulin concentrations were estimated in jugular blood samples from 18 Alpine×Beetal and Sannen×Beetal goats during pregnancy and compared with samples from non-pregnant goats and from goats during the periparturient period. The blood glucose levels in the pregnant goats rose to a peak of about 60±1.36 mg/ml at 42–56 days and then declined to about 46±2.37 mg/ml at 112–126 days. In non-pregnant goats, the blood glucose levels were significantly (p<0.01) higher than in pregnant goats, except between days 42 and 70 (59±1.36 mg/ml). On the day of kidding, the levels declined significantly (p<0.01), increasing again thereafter. The plasma NEFA concentrations were significantly higher in pregnant than in non-pregnant goats from days 56 to 126. The NEFA concentration increased on the day of kidding, followed by a transient fall by day 3. The plasma insulin concentration was usually higher in pregnant than in non-pregnant goats, except between days 56 and 70 and from day 126 onwards. The insulin concentration fell late in pregnancy, but there was a transient increase 2 days after parturition. The blood glucose and plasma NEFA concentrations can be used as indices of nutritional status during pregnancy in goats.     

The Activity of Matrix Metalloproteinase-2 and -9 in Serum of Pregnant and Non-Pregnant Bitches

The aim of this study was to investigate, whether the activity of matrix metalloproteinase (MMP)-2 and -9 in the serum of pregnant and non-pregnant bitches differs significantly. For this purpose, 81 blood samples were taken from pregnant bitches at days 5-13, 15-21, 24-31, 34-40 and 41-50 after mating, and 51 samples from non-pregnant animals at corresponding times. Relative enzyme activity, calculated as the percentage of serum enzyme activity on enzyme activity in a control sample, was determined with a commercially available assay after activation of serum MMPs with p-aminophenylmercuric acetate (APMA). In addition, serum oestradiol (E(2)) and progesterone (P(4)) concentrations were measured with an enzymeimmunoassay (EIA). In the pregnant bitches, at days 5-13 and 15-21 after mating, the mean activity of both MMPs was significantly higher than in non-pregnant animals (28.5% vs 24.5% and 27.7% vs 22.6%; p < 0.01). Moreover, in the pregnant bitches, significant correlations were detected between the serum enzyme activity and the serum concentrations of E(2) (-0.900; p < 0.05) and P(4) (+0.667; p < 0.05).     

Expression of MHC-I and -II in Uterine Tissue from Early Pregnant Bitches

The aim of the study was to investigate the expression of major histocompatibility complex (MHC)-I and -II in uterine tissues from pregnant and non-pregnant bitches, taken at different time periods after mating. The pregnant bitches were ovariohysterectomized during the pre-implantation (group 1, n = 4), implantation (group 2, n = 7) and placentation stage (group 3, n = 7). Non-pregnant animals in diestrus served as controls (group 4, n = 7). The expression of MHC- I and -II in salpinx, apex, middle horn, corpus uteri and at implantation sites was investigated by immunohistochemistry as well as qualitative and quantitative RT-PCR; MHC-I mRNA was detected in all tissues and with quantitative RT-PCR, and no significant changes were detected until placentation. Immunohistologically, at the apex and corpus site, the average number of MHC-II positive cells increased from the pre-implantation to the post-implantation stage (apex: 1.54 ± 1.21 to 3.82 ± 2.93; corpus: 1.62 ± 1.9 to 5.04 ± 4.95; p < 0.05). The greatest numbers of MHC-II positive cells were observed at placentation sites (6.64 ± 5.9). In parallel, a marked increase in the relative mRNA expression of MHC-II in uterine tissues was assessed from the pre-implantation to the placentation stage (relative to Glycerinaldehyd-3-phosphate-Dehydrogenase (GAPDH): 6.9 ± 9.5, 8.4 ± 5.8, p > 0.05). Immunohistologically, in the salpinx, significantly greater numbers of MHC-II positive cells were found in the tissues of pregnant animals than in the control group (p < 0.05). It is proposed that the increase in MHC-II is pregnancy-related, even though the impact on maintenance of canine pregnancy is still unclear.     

Expression and Activity of Matrix Metalloproteinases in the Uterus of Bitches After Spontaneous and Induced Abortion

Aim of this study was to determine the intrauterine activity of matrix metalloproteinases (MMP)‐2 and ‐9 after cessation of the local effect of progesterone. For this purpose, pregnancy was terminated in 10 bitches at mid‐gestation with the progesterone receptor antagonist aglepristone (10 mg/kg body weight, sc, Alizine^®; Virbac, France) at two subsequent days (group IRA = induced resorption/abortion). The IRA group was divided into two subgroups (Group I, n = 5, days 25–35 of pregnancy; group II, n = 5, days 36–45). Five further bitches were introduced with beginning abortion (group SRA = spontaneous resorption/abortion). Seven healthy bitches between day 25 and 45 of gestation served as controls. After ovariohysterectomy at the end of abortion and between days 25 and 45 of gestation, respectively, the distribution and activity of collagenases were investigated by immunohistochemistry and gelatin zymography. At placental sites, MMP‐2 activity in the endometrium was significantly lower in IRA groups than in the SRA group (33.7 ± 11.8% and 39.3 ± 5.4% vs 52.2 ± 10.2%, p < 0.05); however, MMP‐2 expression was lowest in the control group (control: 21.4 ± 6.3%; p < 0.01) and similarly in the myometrium (controls: 13.1 ± 2.5%; p < 0.05). MMP‐9 activity was also lower in the endometrium and myometrium of the control group in comparison to SRA and IRA groups (11.8 ± 3.2%; p < 0.01 and 28.4 ± 32.8%; p < 0.05). At interplacental sites, the amount of active collagenases in the myometrium was significantly lower in the control group. It is concluded that the blockade of the biological progesterone effect was associated with an increase in activity of both collagenases.     

超声断层扫描观察苏北犬胚胎和胎儿发育

用5MHz线阵探头对14只母犬从配种后18d开始，每2d用B超探查1次；配种后36～48d，每3d探查1次；48d以后，于52、57d各探查1次。结果表明，于配种后18～22d确诊7只犬妊娠，7只空怀。有3只犬分剐在配种后24、36、39d发生流产。观察胎囊、胎儿、胎心、胎动、胎儿脏器、胎儿骨骼出现时间和测量胎囊大小、冠臀长、胎儿身长、头长、双顶径、胸深和心区大小，证明胎囊大小、胎儿冠臀长、身长、头长、双顶径、胸深与胎龄呈显著正相关。在配种后18～30d，观察子宫暗区大小可预测胎龄。在配种32d以后，观测胸深、双顶径或头长。可精确预测胎龄。     

妊娠早期绵羊子宫和胎儿实测与超声探查比较

用Ｂ超跟踪探查的９头妊娠母羊，分别于妊娠５７、５８、５９、６０、８６、８７、８８、９６和１００ｄ剖检，实测其于叶直径、胎儿冠一臀长、头长和胎儿主要内脏。结果发现，直径最大的子叶在妊娠８６ｄ，说明以测子叶直径来预测胎龄，只适用于妊娠８６ｄ以前；妊娠５９ｄ观察到有胃液，妊娠８６、９６ｄ先后在直肠和结肠观察到有胎粪蓄积，９６和１００ｄ在羊水和胃内发现有小粪球，说明此时已有排粪和吞咽反射；妊娠８７和８８ｄ观察到膀胱有尿蓄积和睾丸降入阴囊。     

母猪妊娠早期血清和胎儿胎水中硫酸雌酮含量

应用放射免疫法测定枫泾（ＦＪ）和长白（Ｌ）青年母猪妊娠２８ｄ左右外周血清和胎儿羊水、尿囊液中硫酸雌酮（Ｅ１Ｓ）的含量；观察了两品种母猪胚胎存活数。结果，ＦＪ和Ｌ母猪血清、胎儿羊水及尿囊液中Ｅ１Ｓ含量（μｇ／Ｌ）分别为５．３±０．９９、８．０±１．０、３８．０±６．７和３．６±０．４６、５．６±０．９９、６１．４±１１．７，均处于较高水平，且两品种间无显著差异。尿囊液Ｅ１Ｓ含量极显著高于羊水和母体血清含量（Ｐ＜０．０１）；胚胎存活数（个），ＦＪ和Ｌ分别为１４．５±１．３和１０．４±１．３；血清Ｅ１Ｓ浓度与其胚胎存活数呈正相关：ｒＦＪ＝０．７４２６（Ｐ＜０．０１），ｒＬ＝０．４７４７（Ｐ＜０．０５）。上述结果表明，母猪妊娠早期孕体雌激素代谢功能很强，且以Ｅ１Ｓ形式存在于血清和胎水中。母猪血清Ｅ１Ｓ浓度不仅反映早期孕体分泌雌激素的功能，而且是预测早期胚胎存活数的有价值指标     

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.     

神经生长因子(NGF)在母牛性腺及性腺外生殖器官中的表达研究

本研究旨在探讨神经生长因子(nerve growth factor,NGF)在母牛卵泡期及黄体期子宫、卵巢、输卵管3种组织中的表达情况。采用FQ-PCR方法和Western blotting技术对成年母牛卵泡期及黄体期的子宫、卵巢、输卵管中的NGF mRNA和蛋白质表达进行了检测。结果表明,NGF mRNA和蛋白质在卵泡期和黄体期的子宫、卵巢、输卵管中均有表达,且卵泡期子宫中NGF mRNA表达量显著高于卵泡期卵巢和黄体期输卵管中NGF mRNA表达量(P0.05),其它组织间无显著差异。     

Lectin‐binding Sites on the Normal and Pathologic Uterus of Sows

The lectin‐binding pattern was compared in the normal and pathological uterus of sows during the ovarian cycle. The following biotinylated lectins were used: Con A, DBA, SBA, PNA, RCA‐I, UEA‐I and WGA. Glycoconjugate labelling showed differences between phases of ovarian cycle and presence of morphologic lesions. Cystic endometrial hyperplasia increased the RCA‐I reaction in the apical region of the glandular epithelium. There was higher intensity of labelling of WGA in the glandular epithelium in uteri with endometritis. In addition, increased Con A binding in the glandular epithelium and mild reduction of UEA‐I reactivity in the glycocalyx of the glandular epithelium were detected in the cases of endometritis. The results of this study show that morphologic alterations modify the sugar pattern in the porcine uterus. These modifications in glycoconjugates may be one of the reasons for decreased fertility in sows.     

烟碱对LPS诱导的兔子宫内膜上皮细胞炎症因子及前列腺素分泌的影响

试验旨在探索α7烟碱乙酰胆碱受体（α7 nicotinic acetylcholine receptor,α7nAChR）的高亲和力激动剂烟碱对脂多糖（lipopolysaccharides,LPS）诱导的兔子宫内膜上皮炎症的作用机制。分离发情后期兔的子宫内膜上皮细胞,用100 ng/mL LPS对细胞进行炎性刺激12 h。用CCK8法检测不同浓度烟碱（5、10和20 μg/mL）对细胞存活率的影响,筛选合适浓度的烟碱进行后续试验。将细胞分为对照组（CON）、LPS、LPS+烟碱、LPS+烟碱+甲基牛扁碱（MLA）（α7nAChR的特异性颉颃剂）组,通过ELISA法检测细胞培养上清液中白细胞介素-1β（interleukin 1β,IL-1β）、IL-6、IL-8、肿瘤坏死因子-α（tumor necrosis factor,TNF-α）、前列腺素E₂（prostaglandin E₂,PGE₂）和前列腺素F_2α（PGF_2α）的含量。试验成功分离兔子宫内膜上皮细胞,且传至第5代仍保持良好的生长状态。CCK8检测结果显示,20 μg/mL烟碱组细胞存活率显著降低（P<0.05）,10 μg/mL烟碱对细胞存活率无显著影响（P>0.05）,所以选择10 μg/mL烟碱进行后续试验。ELISA结果显示,与对照组相比,LPS组IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量显著增加（P<0.05）;与LPS组相比,LPS+烟碱组显著降低IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α的含量（P<0.05）,LPS+烟碱+MLA组炎性因子和前列腺素的含量差异不显著（P<0.05）。以上结果表明,烟碱对LPS诱导的兔子宫内膜上皮细胞分泌IL-1β、IL-6、IL-8、TNF-α、PGE₂和PGF_2α具有下调作用,推测烟碱通过α7nAChR介导炎性因子和前列腺素分泌下调而发挥抗炎作用,该结果可为研究α7nAChR作为子宫内膜炎治疗靶点的药物选择提供参考。     

Post-Partum Involution of the Canine Uterus – Gross Anatomical and Histological Features

We aimed to study the normal puerperium in the bitch. Ovariohysterectomy was performed in nine bitches, each at a different day after normal whelping; their genital tract was subject to gross anatomical examination, as well as to histological examination and electron microscopy scanning. Corpora albicans were evenly distributed in the left and right ovaries and placental sites were evenly distributed among left and right uterine horns. Placental sites were initially of dark green to grey colour, later becoming dark brown; their length and height progressively decreased. Height of the myometrium and diameter of the uterine glands progressively decreased. Trophoblast-like cells were consistently observed at the placental sites and on the surface of the interplacental areas, at all time points where hysterectomy had been performed. It is suggested that involution of the canine genital tract can last up to 3 months and is slow. Continuous (up to D84 post-partum) presence of prominent placental sites should be considered a normal feature of canine uterine post-partum involution.     

Successful Treatment for Subinvolution of Placental Sites in the Bitch with Low Oral Doses of Progestagen

Subinvolution of placental sites (SIPS) is the major cause of persistent sanguineous vaginal discharge after parturition in the bitch. Spontaneous remission is common but may take several months, and hence, medical therapy to end the discharge is often requested. In this retrospective study, we evaluated the effect of treatment for SIPS with low oral doses of a progestagen. Nine bitches with SIPS, but otherwise clinically healthy, were found in the computer database of the Department of Clinical Sciences of Companion Animals. Seven of these bitches were treated with low oral doses of a progestagen (megestrol acetate, 0.1 mg/kg body weight (bw) once daily for the 1st week, then 0.05 mg/kg bw once daily for the 2nd week). The other two bitches were untreated. Treatment results were evaluated by a telephone questionnaire. Progestagen treatment was successful in all of the treated dogs; sanguineous vaginal discharge stopped within the treatment period. One of the two untreated dogs remained symptomatic until the next oestrus, approximately 120 days after parturition, and the other remained symptomatic until 6 weeks before the start of the next pro‐oestrus, 270 days after parturition. No side effects of the progestagen treatment were observed. Subsequent gestations, parturitions and puerperal periods of 5 mated bitches were uneventful. One bitch did not become pregnant after mating. In conclusion, the results of this study indicate that oral administration of low doses of progestagen for 2 weeks is effective in stopping persistent sanguineous vaginal discharge in bitches with SIPS, with neither side effects nor reduced subsequent fertility.     

Comparison of Two Doses of the GnRH Antagonist, Acyline, for Pregnancy Termination in Bitches

Gonadotropin-releasing hormone (GnRH) antagonists are particularly useful when a rapid inhibitory effect on the gonadal axis is required. The aim of this study was to test the efficacy and clinical safety of a low and high dose of the third generation GnRH antagonist, acyline, on pregnancy termination in female dogs. The effect of the antagonist on the progesterone (P₄) serum concentration was also described. Twenty-one mid-pregnant bitches were randomly assigned to a single subcutaneous (SC) dose of a placebo (PLACE; n = 7), a low (ACY-L; 110 μg/kg; n = 6) or high (ACY-H; 330 μg/kg; n = 8) dose of acyline. The animals were followed up for 15 days. All ACY treated but no placebo-treated animals terminated their pregnancy by abortion (p < 0.01). The ACY-L and ACY-H groups interrupted their pregnancy 7 ± 1.9 and 6.4 ± 1.3   days after treatment, respectively (p = 0.7). A significant interaction between treatment and day was found (p < 0.01) for P₄ serum concentrations when PLACE was compared with both ACY groups. No difference was found for this hormone between both ACY groups (p > 0.05) where P₄ diminished throughout the study. The decreasing rate varied among animals and was closely related to the time of abortion when P₄ reached basal concentrations. In PLACE animals, gestation progressed normally and P₄ did not change throughout the study (p > 0.05). None of the bitches presented side effects. It was concluded that acyline safely terminated mid-pregnancy by permanently decreasing P₄ serum concentrations.     

小鼠子宫壁内自然杀伤细胞和T细胞亚群与自发性胚胎丢失相关性研究

免疫组化研究显示，小鼠妊娠早期子宫壁内CD57^ 细胞数量由孕0天时16.3增至孕2天时62.3,孕4天时已下降，孕6天时降至9.7，而发生胚胎丢失的个体孕6天和孕8天时细胞数量均维持较多，约是孕0天时的4倍。CD4^ 细胞在妊娠后数量由孕0天时29.7增至孕4天时65.6，孕6天时降至7.2，发生胚胎丢失的个体孕6天及孕8天时数量均较多，约为孕0天时的6倍。CD8^ 细胞妊娠后数量由孕0天时39.8增至孕2天时185.0，孕4天时已下降，孕6天时降至3.6，而发生胚胎丢失的个体孕6天和孕8天时细胞数量均维持较多，约是孕0天时的3倍。研究结果提示小鼠妊娠早期子宫壁内CD57^ 细胞、CD4^ 细胞和CD8^ 细胞数量在胚胎着床前均出现先增后减的波动，早期胚胎丢失与孕6天及孕8天时子宫壁内所含大量CD57^ 细胞、CD4^ 细胞和CD8^ 细胞之间存在相关。     

功能性氨基酸改善妊娠和哺乳母猪及新生仔猪的生长与健康状况的研究进展

胎儿及新生仔猪的生长和健康直接受到母猪营养生理状况的影响.妊娠期的限饲和哺乳期采食量的降低均可导致母体处于分解代谢状态,从而引起胎儿生长及乳腺发育中饲粮蛋白质供应不足.由于功能性氨基酸(精氨酸、支链氨基酸、谷氨酸、色氨酸和脯氨酸)能提高受孕率,促进胚胎形成,加速血液流动,增强抗氧化活性,促进食欲,加快蛋白质翻译启动,促...