腧穴电特性与生殖内分泌激素关系研究的初步报告

经络学说认为,经穴是机体脏腑之气输注并散发之部位。脏腑的机能状态,通过经络反映到体表经穴。实验证明,根据经穴电特性的变化,可判断脏腑的机能状态。本试验应用XXH-ⅡA型经穴诊断治疗仪,通过测记70例发情正常马属动物卵泡发育过程中经穴阻抗的变化,对与卵巢机能相关的左右同名经穴的阻抗失衡情况及其与生殖内分泌激素的相互关系进行了研究。试验观察到在卵泡发育前期(1、2、3期),雁旁穴阻抗失衡,卵泡发育后期(4、5期)至排卵后24小时内肾旁穴阴抗失衡。且随卵泡发育成熟的排卵,雁旁穴阻抗失衡率递减、而肾旁穴阻抗失衡率递增。这种变化趋势与血浆中雌二醇、孕酮含量变化趋势相一致。在相关经穴低阻敏感点上给予无针电脉冲刺激,可迅速促使动物自身调节释放孕激素。 试验结果表明:卵巢机能的变化可引起相关经穴阻抗失衡;相关经穴与生殖内分泌激素有着内在联系。这将为经穴与生殖生理、繁殖育种等提供相关的科学依据和研究手段。     

The Effect of PGF2α-Induction of Estrus on Pregnancy Rates in Mares

In mares, the onset of estrus is routinely induced after a luteolytic dose of a prostaglandin F2alpha analogue. Mares in diestrus with a mature, functional corpus luteum will respond by coming into estrus 3 to 4 days after induction¹. Shortening the interestrous interval in mares has an important economic impact on the equine breeding industry. Because the breeding season of the mare is short, timing of insemination and appropriate coordination of endocrine events is critical to pregnancy success and ultimate foaling rates. Several recent studies have reported that the use of prostaglandin analogues is associated with lower pregnancy rates. In this study the induction of estrus with cloprostonol (125-250 ug IM) did not affect pregnancy rates in mares that have ovulatory cycles (n = 461).     

蒙古利亚矮种母马垂体促性腺激素的血清含量与妊娠状态和年龄的相关性

选用 10匹 4～ 16岁怀孕和 2 2匹 0 5～ 17岁非怀孕的蒙古利亚矮种母马 ,采用放射免疫法分别测定了其血清中FSH和LH的浓度。其结果显示 ,妊娠母马的血清FSH和LH含量分别为 0 3 0 83mIU /ml± 0 1611mIU/ml和 0 65 67mIU /ml± 0 40 61mIU/ml,显著低于非妊娠母马的 2 40 5 9mIU/ml± 0 64 46mIU /ml和 2 3 85 5mIU /ml± 0 3 5 2 9mIU/ml(P <0 0 5 ) ;此外 ,3～ 8岁组、10～ 14岁组和 15岁以上组非妊娠母马血清FSH和LH含量分别为 2 44 0 0mIU /ml±0 780 0mIU /ml、2 40 0 0mIU/ml± 1 3 0 78mIU /ml、5 3 75 0mIU/ml± 0 885 0mIU /ml和 1 840 0mIU/ml± 0 80 0 0mIU /ml、2 5 92 9mIU/ml± 0 5 93 0mIU/ml、3 975 0mIU /ml± 0 485 0mIU/ml。以上结果表明 ,在妊娠阶段 ,母马血清中的FSH和LH显著降低 ,而繁殖季节的非妊娠母马血清FSH和LH的浓度却随着母马的老年化而呈现增加的趋势。     

Characterization of Glucose Response Curves after Insulin Injection in Sensitive versus Insensitive Mares

The glucose responses to intravenous injections of a range of doses of recombinant human insulin were determined for six mares known to be insulin sensitive and six mares known to be insulin insensitive, with the goal of better characterizing the regression lines resulting from the two categories of mares. Insulin doses between 8 and 198 mU of insulin per kg of body weight (mU/kg BW) were administered intravenously between September 13 and 26, 2010, starting with 50 mU/kg BW on the first day. Higher and lower doses were administered on alternate days to obtain percentages of decreases in blood glucose concentrations between 10% and 70%. Linear regression analysis revealed that insulin-insensitive mares have glucose response curves with higher y intercepts (P = .066), less steep slopes (P = .0003), and less goodness of fit (P = .053) in addition to the expected greater dose required to produce a 50% reduction in blood glucose concentrations (ED50; P = .006), despite the similarities between their body weights and those of insulin-sensitive mares. Linear and nonlinear regression of responses to the 32, 50, and 79 mU/kg BW insulin doses with the overall estimates of ED50 and the natural log of ED50 indicated that the 50 mU/kg BW dose had the greatest coefficient of determination (>0.95). Generally, it appears that estimates of insulin sensitivity based on a single injection of insulin or on multiple injections of insulin are least variable for insulin-sensitive mares.     

Inhibitory Effects of Pergolide and Cabergoline Formulations on Daily Plasma Prolactin Concentrations in Geldings and on the Daily Prolactin Responses to a Small Dose of Sulpiride in Mares

Two experiments were conducted to assess the efficacy and duration of action of two dopaminergic compounds, pergolide and cabergoline, on daily prolactin secretion in geldings and on prolactin responses to a small dose of sulpiride over 10 days. In the first experiment, oral administration of 2 mg of pergolide was compared to a single injection of 2 mg of pergolide in a slow-release vehicle and a single injection of 5 mg of cabergoline in slow-release vehicle. Controls received vehicle only. All drug treatments reduced (P < .05) prolactin concentrations relative to that in controls but differed substantially in duration of action (oral pergolide approximately 6 hours or less, injected pergolide 6 to 24 hours, and injected cabergoline at least 6 days). In the second experiment, repeated small doses of sulpiride (2 μg/kg of body weight intravenously) were used to stimulate prolactin release in mares, and the ability of seven daily injections of pergolide (2 mg each) and a single injection of cabergoline (5 mg) in slow-release vehicle to suppress this release were compared. Control mares receiving vehicle injections had robust prolactin responses to the sulpiride injections on all days of injection (days 1, 0, 1, 2, 3, 4, 6, 8, and 10 relative to treatment). Prolactin responses were muted (P < .05) by pergolide and cabergoline treatments on the first day of injection (day 0, 30 min after treatment) and were basically absent on days 1 to 8. The single injection of cabergoline continued to be suppressive through day 10, whereas mares previously treated with pergolide (through day 6) had begun to recover a prolactin response by day 10. We conclude that either daily 2-mg pergolide injections in slow-release vehicle or a single injection of 5 mg of cabergoline in slow-release vehicle is an effective way to apply dopaminergic activity to horses for approximately 7 to 10 days and may have application in the treatment of pituitary pars intermedia dysfunction in affected horses.     

Repeatability of Prolactin Responses to a Small Dose of Sulpiride in Estrogen-Primed Geldings in Spring and in Mares During the Estrous Cycle in Summer

Two experiments were conducted to assess the repeatability of prolactin responses to a small dose of sulpiride in estrogen-primed geldings in spring and in mares during the estrous cycle in summer. Six long-term geldings each received a single intramuscular injection of 100 mg of estradiol cypionate on March 31, 2011, and were then challenged with an intravenous injection of dl-sulpiride (5 μg/kg of body weight of the racemic mixture) every other day for a total of 8 days. Jugular blood was collected at 0, 10, 20, 40, and 60 minutes after the injection of sulpiride for prolactin measurement. The experiment was repeated with six mares during the summer (July), except that the number of challenges was extended to 15 over 30 days so that any effect of estrous cycle stage could be assessed. Prolactin responses in geldings during April were robust and were varied in a quadratic manner (P < .003) over the eight sulpiride injections, increasing linearly to a plateau by the fourth injection. Mares also displayed robust prolactin responses to sulpiride injections in July, and there was no effect (P > .1) of day of injection and no effect of stage of estrous cycle (follicular phase, early diestrus, or late diestrus). We concluded that prolactin responses to this dose of sulpiride were sufficiently robust and repeatable for use as a paradigm for studies of the relative competitive efficacy and duration of action of various dopaminergic compounds and their vehicular formulations.     

不同胚胎移植方式的移植效率比较

在人源化抗体转基因猪的研究工作中,将所设计的转基因胚胎成功植入能繁发情受体母猪的输卵管内是推进后续研究工作的基础。所以,本研究对比了两种胚胎移植方法对62例受体母猪的手术效果,结果表明,采用输卵管伞部移植方式的移植效率（60%）显著高于刺破输卵管移植方式（31.8%）。     

Stimulation of Ovulation in Seasonally Anovulatory and Vernal Transitional Mares with Estradiol and Domperidone: Dose and Combination Studies

Three experiments were conducted to test the efficacy of different doses of estradiol cypionate (ECP) and domperidone for inducing ovulation in seasonally anovulatory (January; Experiments 1 and 2) and transitional period (March; Experiment 3) mares. In the first two experiments, mares in Kentucky and Louisiana were administered domperidone (3 g in biodegradable particles) alone or after pretreatment with 100 or 150 mg of ECP; another group received ECP, domperidone, and progesterone, and a fifth group received ECP and progesterone only (the latter two in Kentucky). Control mares in both states received no treatment. The proportion of mares ovulating within 35 days (for mares treated in January) was greater (P = .0002) for those receiving any combination of ECP plus domperidone relative to mares not receiving the combined treatment. Addition of progesterone to ECP plus domperidone did not enhance (P = .7) the response relative to the combination alone; domperidone by itself, or ECP plus progesterone, did not alter the response relative to controls (P > .24). Experiment 3 was conducted in Louisiana as a 2 × 3 factorial, with two doses of domperidone (1.5 or 3 g) and three doses of ECP (0, 75, or 150 mg). There was no main effect of domperidone or ECP dose; a greater proportion (P = .055) of mares receiving any combination of ECP plus domperidone ovulated in 21 days compared with those receiving no ECP. In conclusion, pretreatment with ECP before injection of domperidone 10 days later can be used to increase the proportion of mares ovulating early in the year; within the limits of the present experiments, there appears to be no difference in doses of ECP of 75, 100, or 150 mg, and no difference in doses of domperidone of 1.5 or 3 g.     

Hyperleptinemia in Mares: Prevalence in Lactating Mares and Effect on Rebreeding Success

Previous research from our laboratory showed that approximately one third of obese, nonfoaling mares displayed a condition of hyperleptinemia coupled with hyperinsulinemia that resembled type 2 diabetes in humans. The current study was performed to evaluate the prevalence of the hyperleptinemic syndrome in lactating mares and its possible impact on their rebreeding success. Additionally, we investigated possible relationships between leptin levels in lactating versus nonlactating mares. In experiment 1, jugular blood samples were collected from 198 lactating mares on two occasions approximately 2 weeks apart. The mares resided on eight farms in Louisiana; breeds included Thoroughbred (n = 86), Quarter Horse (n = 71), Warmblood (n = 24), and draft-type (n = 17). Body condition scores (BCS) were measured at the time of blood sampling; plasma samples were assessed for leptin and progesterone concentrations. Reproductive and medical histories, as well as feeding regimens, were compiled on each mare. Based on our previous reports and examination of the current data, a mare was considered hyperleptinemic if her plasma samples contained greater than10 ng/mL leptin; normal was considered 6.0 ng/mL or less; mares with levels above 6.0 and 10 ng/mL or greater were classified as intermediate. Overall mean leptin concentration was 4.7 ng/mL, and average BCS was 5.5. After analysis, 24 mares were classified as hyperleptinemic (12%), 138 were classified as normal (70%), and 36 were classified as intermediate (18%). Leptin concentrations were affected by BCS (P = .08), with higher concentrations in mares with higher body condition; however, there were hyperleptinemic mares with BCS of 4 to 5.5. Feeding regimen affected leptin concentrations (P < .01), with mares on pasture full-time having the highest concentrations. There was no effect of breed, mare age, number of years the mare had been bred, number of live foals, progesterone concentrations, or last foaling date on leptin concentrations. Rebreeding success averaged 81% overall and was not affected by leptin classification. In experiment 2, nonfoaling mares kept on pasture had mean leptin concentrations of 7.0 ng/mL; 8 of 31 mares (26%) displayed hyperleptinemia. Mean leptin concentration was correlated with BCS (R² = 0.65; P < .02) but was not affected by age of the mare. It was concluded that the hyperleptinemic condition occurs in lactating broodmares, even at BCS as low as 4. The overall incidence appears to be lower in broodmares than in nonfoaling mares, likely because of their lower BCS in general and the energy demands of lactation. Hyperleptinemia did not affect rebreeding success at the end of the breeding season.     

The influence of late pregnancy and lactation on bone metabolism in mares

Pregnancy and lactation are periods of significant influence on bone metabolism that has not been investigated in equines. To examine the influence of late pregnancy and lactation on bone metabolism in mares, the changes in the blood serum/plasma total calcium (Ca), inorganic phosphates (Pi), pyridinoline (Pyd) and 17β-estradiol (E2) concentration and the bone alkaline phosphatase (BAP) activity were investigated. The samples were taken from 11 mares on 60 ± 10 and 20 ± 10 days before foaling, and 20 ± 10 and 60 ± 10 days after foaling. The concentration/activity of Ca, Pi, Pyd and BAP increased significantly in early lactation, but the Pyd than decreased in the 4th period. A significant correlation was observed between the E2 and bone metabolism parameters. The results indicate low maintenance of normocalcaemia with reduced bone synthesis in late pregnancy and prove the role of estradiol in bone metabolism in mares during pregnancy and lactation.     

The influence of age on bone metabolism in mares during late pregnancy and lactation

The concentrations of carboxyterminal cross-linked telopeptide of type I collagen (CTx), osteocalcin (OCN), parathyroid hormone (PTH) and insulin-like growth factor (IGF-I) were measured in the blood serum of two age groups of mares during late pregnancy (70–50 days before foaling), during early lactation and at the peak of lactation (10–30 and 55–80 days after foaling, respectively). During late pregnancy, the PTH was higher in older (8–19 years old), compared to younger animals (3.5–4 years old) (P < 0.05). The OCN was higher in younger group during late pregnancy (P < 0.05). IGF-I was higher in the younger group during early lactation, in comparison to late pregnancy and the peak of lactation (P < 0.05). IGF-I did not differ between two age groups of mares. The results indicate on the differences in adaptation of bone metabolism to late pregnancy and lactation in older animals, in comparison to younger animals, reflected by elevated PTH secretion.     

Incidence, Endocrinology, Vascularity, and Morphology of Hemorrhagic Anovulatory Follicles in Mares

The incidence of hemorrhagic anovulatory follicles (HAFs) is approximately 5% and 20% of estrous cycles during the early and late ovulatory season, respectively. The structures are more common in old mares (eg, >20 years), tend to occur repeatedly in individuals, and occur most frequently during the late follicular phase. In a recent study, the day of ovulation in controls and the first day of HAF formation, as indicated by cloudiness of follicular fluid, were defined as day 0. On day -1, future ovulating and HAF groups did not differ in follicle diameter or in the frequency of discrete gray-scale ultrasonic indicators of impending ovulation; however, in future HAFs, a greater percentage of the circumference of the follicle exhibited color-Doppler signals of blood flow. No differences were found between the two groups in systemic concentrations of progesterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) on days -4 to 2, but estradiol was elevated in the HAF group on day -3. The wall of the HAFs developed well-vascularized luteal tissue as indicated by echotexture and color Doppler signals and by the production of near normal levels of progesterone. In conclusion, HAFs formed from viable preovulatory follicles that did not differ from ovulatory follicles in diameter or gray-scale echotexture. Estradiol concentrations were elevated a few days before the failure of ovulation, and the wall of the follicle was more extensively vascularized on day -1.