妊娠高血压外周血中促Th2 的细胞因子水平及IL-2/IL-10 平衡 的临床意义

目的:检测妊娠高血压患者外周血中促Th2的分子IL-4、IL-2与IL-10的水平,探讨IL-2/IL-10在妊高症中的临床意义。方法:选择40例未妊娠妇女为对照组,30例正常妊娠妇女为妊娠组,28例妊娠高血压患者为妊娠高血压组,ELISA检测血清中IL-4、IL-2和IL-10的水平。结果:与对照组外周血中IL-4水平(0.53±0.04)pg/ml相比:正常妊娠组IL-4水平升高至(0.91±0.03)pg/ml(P<0.05),妊娠高血压组IL-4水平(0.67±0.35)pg/ml升高但明显低于正常妊娠组(P<0.01)。与对照组外周血中IL-2水平(0.41±0.05)pg/ml相比:正常妊娠组IL-2水平升高至(0.82±0.11)pg/ml(P<0.01);妊娠高血压组IL-2水平高达1.57±0.22(pg/ml)明显高于其它两组(P<0.01)。妊娠高血压组外周血中IL-10水平明显低于正常妊娠组IL-10水平(P<0.01);妊娠高血压组外周血中IL-2/IL-10比值明显高于于对照组及正常妊娠组的比值。结论:妊娠高血压患者外周血中细胞因子IL-2和IL-10分泌异常且诱导Th2细胞产生的IL-4降低,打破Th1/Th2平衡,致使Th1型免疫反应增强,使早孕期滋养细胞受到免疫损伤以致侵入能力下降,导致妊娠期高血压疾病的发生。     

妊娠高血压外周血中促Th2的细胞因子水平及IL-2／IL-10平衡的临床意义

目的：检测妊娠高血压患者外周血中促Th2的分子IL-4、IL-2与IL-10的水平,探讨IL-2／IL-10在妊高症中的临床意义。方法：选择40例未妊娠妇女为对照组,30例正常妊娠妇女为妊娠组,28例妊娠高血压患者为妊娠高血压组,ELISA检测血清中IL-4、IL-2和IL-10的水平。结果：与对照组外周血中IL-4水平（0．53±0．04）pg／ml相比：正常妊娠组IL-4水平升高至（0．91±0．03）pg／ml（P〈0．05）,妊娠高血压组IL-4水平（0．67±0．35）pg／ml升高但明显低于正常妊娠组（P〈0．01）。与对照组外周血中IL-2水平（0．41±0．05）pg／ml相比：正常妊娠组IL-2水平升高至（0．82±0．11）pg／ml（P〈0．01）;妊娠高血压组IL-2水平高达1．57±0．22（pg／m1）明显高于其它两组（P〈0．01）。妊娠高血压组外周血中IL-10水平明显低于正常妊娠组IL-10水平（P〈0．01）;妊娠高血压组外周血中IL-2／IL-10比值明显高于于对照组及正常妊娠组的比值。结论：妊娠高血压患者外周血中细胞因子IL-2和IL-10分泌异常且诱导Th2细胞产生的IL-4降低,打破Th1／Th2平衡,致使Th1型免疫反应增强,使早孕期滋养细胞受到免疫损伤以致侵入能力下降,导致妊娠期高血压疾病的发生。     

猪蛔虫感染对小鼠肺部免疫病理反应的动态观察

目的:动态观察感染猪蛔虫后,小鼠肺组织的病理变化及肺泡灌洗液中相关细胞因子的变化,从而了解蛔虫感染对肺脏的侵害过程。方法:温箱孵育猪蛔虫受精卵至含蚴卵,用灌胃法感染小鼠,分别在感染后第0、7、14、45天处死小鼠,观察肺组织的病理改变,并通过ELISA方法检测肺泡灌洗液（bronchoalveolar lavage fluid,BALF）中IL-4、IFN-γ、IL-10和TGF-β₁含量的动态变化。结果:感染14天组,肺组织的病理改变最为严重:大量炎症细胞浸润,嗜酸性粒细胞增多,严重者支气管狭窄,肺泡塌陷;感染45天组,小鼠肺组织病理变化比前者显著减轻。BALF中IL-4含量在感染7天、14天后分别为（169.20±34.61pg/ml）,（381.33±57.39pg/ml）,均明显高于未感染组（38.03±6.09 pg/ml）;在感染45天后降低（98.49±25.33 pg/ml）,但仍高于未感染组。IL-10水平在感染后降低;但感染45天组,IL-10水平却有所增高（179.78±21.33 pg/ml）,并高于感染前。IFN-γ、TGF-β₁含量在感染后也有明显降低。结论:猪蛔虫感染初期,小鼠肺部损伤严重,小鼠BALF中促炎症细胞因子占主导地位;但在感染后期,BALF中促炎症细胞因子含量降低,炎症抑制因子IL-10有所增高,炎症缓和。     

孕康口服液对肾虚-黄体抑制型先兆流产大鼠的安胎作用

目的:孕康口服液为已上市中成药,为进一步评价其药效,本实验通过建立肾虚-黄体抑制型先兆流产模型,观察孕康口服液的安胎作用。方法:60只妊娠大鼠随机分为正常对照组(NC),模型组(MG),地屈孕酮组(DT,3.02 mg/kg),孕康口服液低剂量组(YK-L,4 ml/kg)、中剂量组(YK-M,6 ml/kg)、高剂量组(YK-H,9 ml/kg),每组10只。自妊娠第1日,每日上午各给药组按规定剂量灌予受试药,NC组、MG组给予等体积的纯化水,连续10 d;每天下午灌胃造模,除NC组给予纯化水外,其余各组按450 mg/kg体质量灌胃羟基脲,连续9 d,第10日按4.0 mg/kg体质量灌胃米非司酮。妊娠第9日,测定各组大鼠背温、抓力、痛阈、自主活动等行为体征;妊娠第11日,各组腹主动脉取血,测定血清雌二醇(E₂)、孕酮(P)、血栓素B₂(TXB₂)水平;摘取卵巢、连胎子宫,观察胚胎个数和直径,计算卵巢、连胎子宫指数。结果:与NC组比较,MG组背温、抓力、痛阈、自主活动次数、胚胎个数、胚胎直径、连胎子宫指数和血清E₂、P、TXB₂水平均显著降低(P＜0.05,0.01)。与MG组比较,孕康口服液各剂量组背温、抓力、胚胎个数、胚胎直径和血清E₂、P水平均显著升高(P＜0.05,0.01);YK-M、YK-H组痛阈、自主活动、连胎子宫指数显著升高(P＜0.05);YK-H组血清TXB₂水平明显升高(P＜0.05)。结论:孕康口服液对肾虚-黄体抑制导致的先兆流产大鼠具有明确的补肾安胎作用,其机制可能与升高血清E₂、P、TXB₂水平,改善肾虚体征和提高胚胎质量有关。     

孕中期巨细胞病毒宫内感染与血Free-β-HCG水平的相关性研究

目的:通过检测孕中期妇女人巨细胞病毒(HCMV)活动性感染、宫内感染以及血Free-β-HCG水平,分析血Free-β-HCG水平与HCMV宫内感染的相关性,初步探讨其可能影响机制.方法:通过酶免法测孕妇血HCMV-IgM和定荧光定量PCR对孕中期妇女血清和羊水中HCMV-DNA的检测以及DILFIA法(时间分辨免疫荧光法)定量检测孕中期妊娠妇女血Free-β-HCG的含量,研究HCMV感染状况与Free-β-HCG之间的相关性,探讨HCMV对母血Free-β-HCG水平的影响.结果:718例样本中共检出HCMV-IgM阳性和(或)HCMV-DNA阳性者共43例并进一步行产前诊断羊水HCMV-DNA阳性14例.孕中期HCMV活动性感染率为5.98％,HCMV宫内感染率为2.22％.孕中期感染组Free-β-HCG含量:7.32± 2.25 ng/ml,非感染组:8.47± 3.17 ng/ml,对照组:10.10± 3.67 ng/ml.结论:HCMV宫内感染组比非感染组孕中期外周血Free-β-hcG的测定水平降低,两者结果之间的差异具有统计学意义.HCMV可通过损伤胎盘绒毛滋养层细胞,使Free-β-hcG的分泌减少.     

复合益生菌活菌制剂对肝硬化患者肠道菌群的影响

目的:观察肝硬化患者口服三联活菌后肠道菌群、血氧及血浆内毒素的变化.方法:选择肠道菌群中具有代表性的细菌进行培养和计数.42例肝硬化患者给予三联活菌治疗21 d.测定治疗前后肠道菌群菌落计数、血氨(干片法)、血浆内毒素(改良鲎试验法).结果:与正常对照组相比,肝硬化组患者存在不同程度的肠道菌群失调,主要表现为双歧杆菌减少(10.12±0.71比9.27±1.25,P＜0.05).治疗后,双歧杆菌由9.27±1.25增至10.43±1.25,差异有显著性(P＜0.01).且血氨水平降低,并可降低肝硬化患者血浆内毒素水平[(109.21±12.23)pg/ml比(71.46±9.12)pg/ml,P＜0.05].结论:肝硬化患者存在肠道菌群失调.益生菌制剂可有效改善肝硬化患者肠道菌群失调,并降低血氨及血浆内毒素.     

人脐带间充质干细胞对小鼠实验性溃疡性结肠炎的疗效

目的探讨人脐带间充质干细胞移植术对溃疡性结肠炎(UC)小鼠的疗效,以及对血清中肿瘤坏死因子α(TNF-α)、白细胞介素-8(IL-8)水平的影响。方法选用雄性小鼠48只,完全随机分为正常对照组、模型对照组、脐带间充质干细胞(hUC-MSCs)治疗组,每组16只。模型对照组与hUC-MSCs治疗组通过口服5﹪葡聚糖硫酸钠诱导UC模型。对hUC-MSCs治疗组小鼠经尾静脉注射脐带间充质干细胞。观察各组疾病活动指数评分;采用酶联免疫吸附(ELISA)法测量各组小鼠血清中TNF-α、IL-8的表达水平,各组数据两组比较采用t检验,多组比较使用重复测量资料的方差分析对结果进行统计分析。结果模型对照组与hUC-MSCs治疗组DAI评分从第2天开始上升,直至实验结束分别为8.72±0.71、4.01±0.35均高于正常对照组0.21±0.20,且差异有统计学意义(F=1 300.66,P0.01);模型对照组与hUCMSCs治疗组比较,在造模期间及治疗的第4天DAI评分分别为7.95±0.71、8.01±0.73,差异无统计学意义(t=-0.24,P=0.08),在治疗的第5天hUC-MSCs治疗组DAI评分开始下降为7.23±0.82,且与模型对照组8.01±0.81相比,差异有统计学意义(t=-2.71,P=0.01)。造模第7天,模型对照组与hU C-MSCs治疗组两组TNF-α分别为(363.32±15.84)pg/ml、(375.52±16.21)pg/ml,均高于空白对照组(112.14±11.23)pg/ml(F=624.12,P0.01);模型对照组与hUC-MSCs治疗组两组IL-8分别为(65.32±10.12)pg/ml、(57.21±8.71)pg/ml均高于空白对照组(25.02±7.81)pg/ml(F=34.29,P0.01),但两组比较差异无统计学意义(P0.05);第2次干细胞注射7 d后即实验结束时,模型对照组与hUC-MSCs治疗组TNF-α分别为(453.23±15.63)pg/ml、(276.84±17.91)pg/ml,仍高于空白对照组(134.32±12.31)pg/ml(F=642.41,P0.01);模型对照组与hUC-MSCs治疗组两组IL-8分别为(75.23±6.72)pg/ml、(39.12±4.42)pg/ml仍高于空白对照组(27.12±5.91)pg/ml(F=113.90,P0.01),但hUC-MSCs治疗组水平较造模结束时有所下降,低于模型对照组(P0.05)。结论间充质干细胞移植术对UC小鼠有良好的治疗作用,且能降低血清中TNF-α、IL-8的水平。     

rhBNP治疗射血分数保留急性心力衰竭的临床研究

目的:探讨重组人脑利钠肽(rh BNP)对急性射血分数保留心力衰竭的临床治疗作用。方法:选择2008年6月至2014年6月间我院收治的20例急性射血分数保留心力衰竭患者为研究对象,其中10例予以硝普钠静脉注射48小时,10例予以rh BNP静脉注射48小时。48小时后,评估和比较两组患者心力衰竭的临床改善情况;72小时后,复查和比较两组患者血浆NT-pro BNP的下降值。结果:硝普钠组患者呼吸困难缓解积分(1.4±1.35)明显低于rh BNP组(3.5±0.71),rh BNP组呼吸困难缓解时间(3.68±1.69 h)显著短于硝普钠组(21.50±18.43 h),血浆NT-pro BNP浓度下降值(864.60±585.92 pg/ml)明显高于硝普钠组(423.30±284.06 pg/ml)。结论:rh BNP治疗急性射血分数保留心力衰竭的临床疗效优于硝普钠。     

游离睾酮指数联合血清GnSAF、SHBG对多囊卵巢综合征不孕患者IVF-ET治疗妊娠结局的预测价值

摘要 目的：探讨游离睾酮指数（FAI）联合血清促性腺激素平抑因子（GnSAF）、性激素结合球蛋白（SHBG）对多囊卵巢综合征（PCOS）不孕患者体外受精-胚胎移植（IVF-ET）治疗妊娠结局的预测价值。方法：选取2020年1月～2022年6月湖南省妇幼保健院生殖医学中心收治的197例PCOS不孕患者为PCOS组,根据IVF-ET治疗妊娠结局分为妊娠失败组和妊娠成功组,另选取同期68名体检健康妇女为对照组。收集PCOS不孕患者临床资料,计算FAI并检测血清GnSAF、SHBG水平。采用单因素和多因素Logistic回归分析PCOS不孕患者IVF-ET治疗妊娠结局的影响因素,采用受试者工作特征（ROC）曲线分析FAI和血清GnSAF、SHBG对PCOS不孕患者IVF-ET治疗妊娠结局的预测价值。结果：PCOS组FAI和血清GnSAF水平高于对照组,SHBG水平低于对照组（P＜0.05）。197例PCOS不孕患者IVF-ET治疗妊娠成功率为51.27%（101/197）。单因素分析显示,妊娠失败组体质指数、黄体生成素（LH）、LH/促卵泡生成素（FSH）、睾酮、抗苗勒管激素（AMH）、FAI、GnSAF高于妊娠成功组,FSH、受精率、优胚率、SHBG低于妊娠成功组（P＜0.05）。多因素Logistic回归分析显示,体质指数增加和LH、LH/FSH、AMH、FAI、GnSAF升高为PCOS不孕患者IVF-ET治疗妊娠失败的独立危险因素,SHBG升高为其独立保护因素（P＜0.05）。ROC曲线分析显示,FAI和血清GnSAF、SHBG联合预测PCOS不孕患者IVF-ET治疗妊娠结局的曲线下面积大于FAI、GnSAF、SHBG单独预测。结论：FAI和血清GnSAF、SHBG水平联合预测PCOS不孕患者IVF-ET治疗妊娠结局的价值较高。     

AMI 患者血浆NT-proBNP 水平与心肌缺血及预后关系的研究

目的:探讨急性心肌梗死(AMI)患者血浆N末端B型尿钠肽前体(NT-proBNP)的水平与心肌缺血及预后的关系。方法:98例急性心肌梗死患者根据患者是否行直接PCI手术治疗,分为PCI手术治疗组和非PCI治疗组,观察缺血改善情况与NT-proBNP水平的关系,同时根据治疗后NT-proBNP的水平分为三组,A组NT-proBNP<125pg/ml、B组125pg/ml≤NT-proBNP<450pg/ml、C组NT-proBNP≥450pg/ml,观察NT-proBNP的水平与预后的关系。结果:行PCI组NT-proBNP的水平下降程度(438.3±134.5)明显高于未行PCI组者(158.6±146.1,P<0.05),MACE的发生情况C组明显高于A组(P=0.006<0.01),也高于B组(P=0.028<0.05),A组与B组相比,B组的MACE发生率有上升的趋势,但是无统计学意义(P=0.432>0.05)。结论:急性心肌梗死患者早期血浆NT-proBNP的水平在一定程度上可以反应心肌的缺血程度,且与患者的预后成明显的负相关。     

多原核受精与HCG日雌孕激素、获卵数、授精密度、受精率、妊娠率的关系

回顾性分析体外受精-胚胎移植(IVF-ET)的686个周期,比较多原核受精组(348例)和非多原核受精组(338例)两组的一般资料、HCG日雌孕激素、获卵数、授精密度及活力、受精率、卵裂率、妊娠率.结果显示两组HCG日雌孕激素、获卵数、成熟卵率、受精率、卵裂率、妊娠率之间有显著差异(P0.05),授精密度、受精活力之间无差异.表明HCG日雌孕激素较高,获卵数多及未成熟和过熟卵子比例高,预示着发生多原核受精的可能性大,而多原核受精的发生可能有较好的临床结局.     

Control of luteolysis in the one-humped camel (Camelus dromedarius)

Blood plasma concentrations of 13,14-dihydro-15-keto PGF2 alpha (PGFM) were measured in groups of mature non-pregnant and pregnant camels to study PGF2 alpha release patterns around the time of luteolysis and the timing of the signal for pregnancy recognition. Injection of each of four camels with 10 and 50 mg of PGF2 alpha showed clearly that five times the dose of exogenous hormone produced five times the amount of PGFM in peripheral plasma, thereby indicating that, as in other animal species, PGFM is the principal metabolite of PGF2 alpha in the camel. Serial sampling of three non-pregnant camels on each of days 8, 10 and 12, and three pregnant camels on day 10, after ovulation for 8 h showed a significant (P < 0.05) rise in mean plasma PGFM concentrations only on day 10 in the non-pregnant, but not the pregnant, animals. A single intravenous injection of 20, 50 or 100 iu oxytocin given to three groups of three non-pregnant camels on day 10 after ovulation did not increase their basal serum PGFM concentrations. However, daily treatment of six non-pregnant camels between days 6 and 15 (n = 3) or 20 (n = 3) after ovulation with 1-2 g of the prostaglandin synthetase inhibitor, meclofenamic acid, inhibited PGF2 alpha release and thereby resulted in continued progesterone secretion throughout the period of meclofenamic acid administration. These results showed that, as in other large domestic animal species, release of PGF2 alpha from, presumably, the endometrium controls luteolysis in the dromedary camel. Furthermore, reduction in the amount of PGF2 alpha released is associated with luteal maintenance and the embryonic signal for maternal recognition of pregnancy must be transmitted before day 10 after ovulation if luteostasis is to be achieved. However, the results also indicate that, in contrast to ruminants, the release of endometrial PGF2 alpha in the non-pregnant camel may not be controlled by the release of oxytocin.     

Follicular fluid levels of vascular endothelial growth factor and leptin are associated with pregnancy outcome of normal women participating in intracytoplasmic sperm injection cycles

Cytokines play a critical and multifarious role in follicular maturation. Consequently, they may influence the pregnancy outcome in cycles of assisted reproduction. The aim of this study was to measure the levels of tumor necrosis factor-alpha (TNFalpha), vascular endothelial growth factor (VEGF) and leptin in serum and follicular fluids (FFs) from women undergoing controlled ovarian hyperstimulation (COH) for intracytoplasmic sperm injection cycles (ICSI). We tried to investigate their interrelationships and to evaluate them as predictive markers for the cycle's outcome. Seventeen women participated in this study. Male factor infertility was the only indication for ICSI cycles. For COH, the long agonist protocol with triptorelin and recombinant FSH was employed. Cytokines levels were evaluated by ELISA. Serum cytokine levels did not differ between pregnant and non-pregnant women. FF-VEGF levels were significantly elevated in non-pregnant women (722.2+/-1093.2 pg/ml) as compared to pregnant women (290.3+/-259.8 pg/ml). Leptin concentrations were also significantly higher in FFs of non-pregnant women (682.6+/-625.1 ng/ml) than those of pregnant women (231.6+/-286.5 ng/ml). There were significant positive correlations between FF-leptin and age, as well as between FF-leptin and FF-VEGF concentrations. It was concluded that elevated FF-leptin and VEGF levels are associated with failure of conception in IVF cycles and may serve as markers in clinical practice.     

Elevated Serum Level of IL-35 Associated with the Maintenance of Maternal-Fetal Immune Tolerance in Normal Pregnancy

Objectives

IL-35 is a novel inhibitory cytokine. In this study, we investigate the serum levels of inhibitory cytokines IL-35, IL-10 and TGF-β in both normal pregnancies and non-pregnant females, and whether IL-35 is associated with the pathogenesis of recurrent spontaneous abortion. We also try to elucidate the relationships of IL-35 with estrogen and alpha-fetoprotein (AFP).

Methods

The levels of IL-35, IL-10, TGF-β, estradiol (E2), unconjugated estriol (uE3) and AFP were analyzed in 120 normal pregnancies, 40 women suffering recurrent spontaneous abortion, 40 postpartum healthy women and 40 non-pregnant women by enzyme-linked immunosorbent assay (ELISA). The correlations between inhibitory cytokines, estrogen and AFP were assessed with the Spearman rank correlation coefficient.

Results

Data are expressed as median and percentiles (Q1, Q3).The level of serum IL-35 in normal pregnancies was significantly higher than that in non-pregnant women [333.6 (59.32, 1391) pg/mL vs. 123.9 (8.763, 471.7) pg/mL; P < 0.001]. A significantly higher level of TGF-β was observed in the first trimester only as compared to non-pregnant women [473.4 (398.0, 580.5) pg/mL vs. 379.7 (311.0, 441.3) pg/mL, P < 0.01]. The difference in serum IL-10 level between pregnant women and non-pregnant women was not significant [8.602 (5.854, 12.89) pg/mL vs. 9.339 (5.691, 12.07) pg/mL; P > 0.05]. The level of serum IL-35 in recurrent spontaneous abortion was significantly lower than that in normal early pregnancy [220.4 (4.951, 702.0) pg/mL vs. 386.5 (64.37, 1355) pg/mL; P < 0.05]. The higher IL-35 level in first trimester pregnant women correlated with E2 (r = 0.3062, P < 0.01) and AFP (r = 0.3179, P < 0.01).

Conclusion

Serum levels of IL-35 increased in normal pregnancy and decreased in recurrent spontaneous abortion. Increased IL-35 correlated with estrogen and AFP levels in early pregnancy. IL-35 is becoming recognized as an active player in the maintenance of a successful pregnancy, but this is not the case for IL-10 or TGF-β.     

Influence of pregnancy on the course of malaria in mice infected with Plasmodium berghei

The course of malarial infection was compared in pregnant mice inoculated with Plasmodium berghei at different stages of gestation. When 12-14 wk old, pregnant BALB/c mice were inoculated with 1 x 10(6) of P. berghei NK65-infected red cells at gestation day 0, 2, 4, 6, 8, 10, 12, 14 or 16, the mice inoculated on gestation days 6-12 expired 6.5 days after inoculation compared to 9.5 days in non-pregnant mice. Parasitemia in these pregnant mice increased rapidly on day 4 after inoculation and anemia also developed earlier on day 5. However, the degree of parasitemia and anemia in the terminal stage of infection in these pregnant mice was milder than that of non-pregnant controls. Blood urea nitrogen increased at the terminal stage although the degree of increase in mice inoculated on gestation days 6-10 was comparatively small. Pregnant malarial mice died earlier with less physiological changes than non-pregnant controls. It was concluded that pregnancy makes the host susceptible to physiological changes caused by malaria.     

Estrone concentration in the peripheral plasma of pregnant and non-pregnant gilts

A rapid radioimmunoassay for estrone (total unconjugated and sulfated) was developed to determine plasma estrone (E1) concentrations in inseminated gilts that conceived and those that had not. Thirty-one 160 day-old prepuberal gilts were induced to ovulate with gonadotropins and were artificially inseminated 10 hr before the expected time of ovulation (Day 1 - day of insemination). Unconjugated E1 and E1SO4 were extracted from 20 to 500 microl of plasma twice with 5 ml of tetrahydrafuran:ethyl acetate (1:1). Aliquots of a standard E1SO4 preparation were dissolved in 500 microl of distilled water and extracted at the same time as the plasma samples. The dried extracts were solvolyzed for 1 hr at 50 degrees C in 0.6 ml of glacial acetic acid:ethyl acetate (1:1), and the dried residue was redissolved in 0.2 ml of distilled water and extracted once with 2 ml of diethyl ether. Twenty of 31 gilts were pregnant at Days 29 to 31 of the induced cycle. Plasma E1 in pregnant gilts increased from 85 pg/ml on Day 18 to 702, 1879 and 2793 pg/ml, respectively, on Days 22, 25 and 29 to 31. Three of the non-pregnant gilts had plasma progesterone secretion maintained until Day 22; they also had a transitory increase in plasma E1 on Day 22 (215 pg/ml). Some blastocysts may have been present to exert a temporary luteotropic effect, but not enough blastocysts to completely overcome the luteolytic effect of the uterus. Quantification of plasma E1SO4 could be used as a pregnancy test in the pig.     

An investigation of the uterine luminal environment of non-pregnant and pregnant pony mares.

Uterine flushings were collected from 30 non-pregnant Pony mares on Days 8, 12, 14, 16, 18 or 20 after oculation. Mares were allowed a recovery period of one oestrous cycle and were mated at the next oestrus. They were then ovario-hysterectomized on days which corresponded to the day of the oestrous cycle to which they were assigned. Uterine flushings were analysed for total recoverable protein and acid phosphatase activity. Least squares analysis indicated a status X day interaction for total protein (P less than 0.10) and acid phosphatase activity (P less than 0.005) in which the latter was higher in uterine flushings during pregnancy. Peripheral plasma oestrone and oestradiol concentrations were measured by radioimmunoassay and results indicated that plasma oestrone concentrations in pregnant and non-pregnant mares were not different, and oestradiol was lower (P less then 0.005) in the peripheral plasma during pregnancy. conceptus membranes were incubated in vitro for 120 min in a chemically defined medium. Incubation medium was then assayed to assess oestrone and oestradiol production capacilities at Days 8, 12, 14, 16, 18 and 20 of pregnancy. Conceptus membrane production of oestradios (pg/5 ml/h) increased (P less than 0.05) from Day 8 (243 pg/5 ml) to Day 20 (108 763 pg/5 ml). A similar trend, but of lower magnitude, existed for oestrone production.     

Changes in cumulus-oocyte complexes of pregnant and non-pregnant camels (Camelus dromedarius) during maturation in vitro

The aim of the present study was to examine the cumulus morphology and the oocyte chromatin quality of camel cumulus-oocyte complexes (COCs) at the time of recovery, and to monitor changes in oocyte chromatin configuration and apoptosis in cumulus cells from camel COCs during in vitro maturation (IVM) (0, 12, 24, 32, 36, 42, and 48 p.IVM) depending on pregnancy of donors. A total of 1023 COCs were isolated from sliced ovaries after slaughtering of 47 pregnant and 43 non-pregnant camels in an abattoir. The mean number of COCs per donor was 10.3 in pregnant and 12.5 in non-pregnant donors. The cumulus morphology of COCs was independent of the type of donor and was divided in COCs with compact (26.9 and 28%), dispersed (39.3 and 46%), corona radiata cumulus investment (27.9 and 21.7%) and without cumulus (6 and 4.2%), respectively for pregnant and non-pregnant donors. The highest proportion of COCs exhibited dispersed cumulus (P<0.05). Oocytes with meiotic stages of diplotene >50% were found only in compact (55 and 56.5%) and in dispersed COCs (58.4 and 60%), respectively for pregnant and non-pregnant donors. During IVM (0-48h) the first significant onset of specific meiotic stages were different in oocytes from pregnant donors: metaphase 1 (24-32h), metaphase 2 (36-42h), versus oocytes from non-pregnant donors: metaphase 1 (24h), metaphase 2 (32-48h) (P<0.05). The level of apoptotic cells in cumuli of matured COCs increased during IVM and was higher in matured COCs from non-pregnant donors for each time point during IVM (P<0.01). Camel oocytes meiosis during IVM is accompanied by a drastic increase of apoptosis in the surrounding cumulus cells 0-32 and 0-24h during IVM, respectively for pregnant and non-pregnant donors. The oocytes of pregnant camels require 36h of maturation to reach levels of >50% metaphase 2 stage in comparison to oocytes from non-pregnant donors where 32h are sufficient. The earlier onset of apoptosis in the COCs derived from non-pregnant donors possibly determines the faster progression of the oocytes through the final stages of meiosis.     

Hormonal contraception of feral mares with Silastic rods.

Homogeneous Silastic rods containing ethinylestradiol (EE) (1.5 or 4 g), estradiol-17 beta (E) (4 g) or progesterone (P) (6 g) were implanted into feral mares (Equus caballus) between 4- and 10-yr-old. Six treatment groups (greater than or equal to 10 mares/group) of non-pregnant mares received 36 g P and 12 g E (P+E), 36 g P and 8 g EE (P+HEE), 1.5 g EE (LEE), 3 g EE (MEE, 8 g EE (HEE) or control-implanted mares (CI). CI received implants containing no steroid. Two groups of pregnant mares received P+HEE or HEE. Stallions were placed with the mares 15 to 26 mo after implanting. Blood was collected biweekly for up to 28 mo after implanting and serum analyzed for P by radioimmunoassay. A single P value greater than or equal to 2.5 ng/ml indicated ovulation and 2 consecutive values greater than or equal to 2.5 ng/ml indicated pregnancy. Serum from blood collected before and at 4, 12, 24, 50, 64 and 89 wk after implanting was analyzed for EE concentrations. All animals pregnant at the time of contraceptive placement delivered normal foals. Contraceptive efficacy for groups LEE, MEE, HEE and P+HEE were 75, 75, 100, and 100%, respectively after two breeding seasons. Suppression of ovulation appeared to be inversely related to the concentration of EE used in the implant. The percent of animals ovulating after 2 yr of contraception in each group was 100, 100, 88, 62, 20, and 12 for groups CI, P+E, LEE, MEE, HEE and P+HEE, respectively. The pregnancy rate for the same groups was 100, 78, 25, 25, 0 and 0%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Circulating oestrogen concentrations during pregnancy in the African elephant (Loxodonta africana)

Oestrone, oestradiol-17 beta and oestriol were measured in plasma samples from non-pregnant and pregnant African elephants shot in the wild. Enzymic hydrolysis of plasma showed that approximately 90 and 96% of the total (i.e. conjugated plus unconjugated) concentrations of oestrone and oestradiol-17 beta, respectively were represented by conjugated hormones. Unconjugated oestrogens remained low (less than 50 pg ml) in all samples, with no distinction between non-pregnant and pregnant animals. Levels of total oestrone during pregnancy varied between 160 and 594 pg/ml but were not significantly different from non-pregnant values. Total oestradiol-17 beta concentrations were significantly elevated during pregnancy (P less than 0 X 01) and, despite considerable individual variation (193-1428 pg/ml), were consistently higher than non-pregnant values after 6 months of gestation. The elevated levels of oestradiol-17 beta resulted in a reversal of the total oestradiol-17 beta: oestrone concentration ratio at about 6 months of pregnancy. Concentrations of total oestriol did not exceed 103 pg/ml. An indirect method of measurement indicated that oestradiol-17 beta sulphate was probably the most abundant circulating oestrogen during pregnancy in the African elephant.