Plasminogen Activator and Plasminogen Activator Inhibitor in Mouse Ovaries during Periovulatory Periods

Changes of ovarian tPA,uPA and PA inhibitor activities were examined in PMSG-and hCG-treatedimmature mice during periovulatory periods.The results show that 15% of the gonadotropin-treatedanimals ovulated 8 hrs after hCG administration,about 6-8 hrs earlier than in rat.It is also shownthat not only tPA activity,but also uPA activity,was regulated by gonadotropins in ovarianhomogenates and granulosa cells,and they reached maximum prior to ovulation.No measurableamount of PAI-1 activity could be detected in mouse granulosa cell conditioned medium andfollicular fluid,but considerable amount of α_2-antiplasmin,a specific inhibitor for plasmin,wasfound in follicular fluid.Cumulus-oocyte complexes contain mainly tPA.Since the ovulated eggsstill have high tPA activity,it is thought that the enzyme in the oocyte may play an important rolein implantation.

Sperm hyaluronidase is critical to mammals'fertilization for its ability to disperse cumulus-oocyte complex layer

Glycosylphosphatidylinositol-anchored sperm hyaluronidases have long been believed to assist in sperm penetration through the cumulus-oocyte complex(COC);however,their role in mammalian fertilization remains unclear.Previously,we have shown that hyaluronidase 5(Hyal5)/Hyal7 double-knockout(dKO)mice produce significantly fewer offspring than their wild-type(WT)counterparts because of defective COC dispersal.Male infertility is mainly caused by a low sperm count.It can be further exacerbated by the deficiency of sperm hyaluronidase,which disperses the cumulus cells of the outer layer of the COC.In the current study,we evaluated the effects of a low count of Hyal-deficient sperm and conditions of ovulated oocytes on the fertilization rate using a mouse model.Our results demonstrated that a low sperm count further decreases the in vitro fertilization(IVF)rate of Hyal-deficient dKO spermatozoa.In addition,the dKO spermatozoa resulted in a fertilization rate of 12.5%upon fertilizing COCs with a thick cumulus layer,whereas the IVF rate was comparable to that of WT spermatozoa when oocytes with a thin or no cumulus layer were fertilized.Finally,we proved that the IVF rate of dKO spermatozoa could be recovered by adding rat spermatozoa as a source of sperm hyal.Our results suggest that a deficiency of proteins involved in fertilization,such as sperm hyal,has a vital role in fertilization.

小鼠排卵前后卵巢纤蛋白溶酶原激活因子活性的变化

给幼龄小鼠注射PMSG刺激滤泡生长,随后注射hCG以诱发排卵。在激素处理的不同时间取出卵巢,制备卵巢匀浆液或从卵巢中分离颗粒细胞和卵丘-卵母细胞复合体,并做离体培养。样品中组织型(tPA)和尿激酶型(uPA)纤蛋白溶酶原激活因子经SDS-凝胶电泳分离,用纤蛋白铺盖技术测定。实验结果表明,注射hCG 8h后15％的受试动物排卵,而卵巢匀浆液和颗粒细胞中tPA和uPA活性分别也在hCG注射后4和8h达到高峰。排卵后酶活性下降。卵丘-卵母细胞复合体主要含tPA,注射hCG 12—24h达到高峰。上述资料证明,tPA和uPA都参入小鼠排卵过程。因为排出的卵子中仍含有大量tPA,卵细胞的tPA除参与排卵外,可能对排卵后的一些生理过程也起重要作用。

基于复杂网络分析中药人工周期治疗排卵障碍性不孕症的用药规律

目的通过现代文献研究分析中药人工周期治疗排卵障碍性不孕症的用药特点,为提高临床疗效提供依据。方法对2006-2015年的相关临床研究文献进行检索,应用Excel软件对不同周期所用中药建立数据库,运用中医复杂分析系统软件对所用中药进行复杂网络分析。结果纳入文献62篇,其中48篇分4期治疗,14篇分3期治疗。经后期治疗的核心药物有:菟丝子、熟地黄、山药、枸杞子、当归、山茱萸、白芍、女贞子、淫羊藿、何首乌;经间期治疗的核心药物有:当归、菟丝子、熟地黄、续断、淫羊藿、川芎、桃仁、红花、赤芍、丹参、香附;经前期治疗的核心药物有:菟丝子、淫羊藿、仙茅、巴戟天、肉苁蓉、当归、白芍、熟地黄、续断、枸杞子、山药;行经期治疗的核心药物有:当归、益母草、川芎、赤芍、香附、桃仁、泽兰、红花、熟地黄。结论通过复杂网络分析中药人工周期治疗排卵障碍性不孕症的用药规律,说明排卵障碍性不孕症病机是肝肾亏虚、血虚和血瘀。分期治疗:经后期补肝肾,滋阴养血;经间期补肝肾,行气活血;经前期补肾温阳养血;行经期以活血化瘀为主。为临床治疗提供了一定依据。

辽宁省牛寄生虫区系调查及其防治的研究

1999年1～12月 ,在辽宁省7市、县设立了31个调查点 ,进行牛寄生虫的区系调查 ,调查了辽宁省牛体内外寄生虫的种类、优势虫种及其消长规律。牛体内寄生虫的优势虫种是肝片形吸虫和消化道圆形线虫 ,体外寄生虫的优势虫种是蜱和螨 ,肝片形吸虫和消化道圆形线虫的排卵高峰期在3月、7月和11月。在排卵高峰期前的2月末、6月末和10月末 ,用阿维菌素以0.2mg·kg-1 体重 剂量口服 ,7d后 ,再用丙硫苯咪唑以15mg·kg-1 体重 剂量口服。用这两种药物复合应用进行驱虫 。

Higher Tolerance of Min Pigs Under Cold Stress:An Assessment from Reproduction Perspective

Raising Min pigs(MPs)in northern China is challenging,because of harsh environmental conditions in winter.There is a hypothesis that MPs can tolerate cold stress better than Large White pigs(LWPs)and their oocyte competence is particularly protected.Over a period of 3 years,multiple cohorts of MPs and LWPs under different environmental conditions were studied in this paper.Follistatin(FST)serum level was used as a marker for swine oocyte competence.Oocytes were collected from the swine and their maturation rate,cleavage rate and blastocyst rate were measured.Lastly,mRNA expression in ovary and longissimus dorsi for the genes FST,luteinizing hormone receptor(LHR)and cold-inducible RNA-binding protein(CIRP)was measured.Under cold stress,serum follistatin concentrations of both LWPs and MPs were markedly increased,and they were more in LWPs than in MPs.However,oocyte competence was found to be unaffected by cold stress in MPs.It was found that in a separate group of LWPs,oocyte competence was lower in samples collected in winter than that in other seasons.Interestingly,follistatin levels in the follicular fluid of MPs showed no change with cold stress,and serum follistatin levels were increased in sows.Gene expression analysis showed a significant reduction in FST,LHR and CIRP expressions under cold stress in longissimus dorsi,but not in the ovaries.It was found that MPs were better able to maintain oocyte competence under cold stress,which might be due to improved regulation of follistatin in follicular fluid,and MPs appeared likely an adaptation to the harsh conditions that they faced under domestication in northern China.

ICR小鼠卵母细胞通过旁分泌机制促进排卵关键过程

目的:利用国内医学研究常用ICR品系小鼠,在构建用于研究卵母细胞-卵丘细胞交互作用培养模型的基础上,研究卵母细胞及其产生的旁分泌因子生长分化因子9(GDF9)和骨形态发生蛋白15(BMP15)对排卵过程中的关键事件——卵丘扩展的调控作用。方法:通过显微手术制备摘除卵母细胞后的卵丘细胞复合体(OOX),并通过与卵母细胞共培养或GDF9和BMP15处理研究卵母细胞及其特异释放的旁分泌因子对表皮生长因子(EGF)诱导卵丘扩展过程的调控。结果 :成功构建了ICR小鼠OOX培养模型,发现卵母细胞及其释放的旁分泌因子GDF9和BMP15为EGF诱导卵丘扩展所必需。结论:ICR小鼠卵母细胞通过旁分泌机制促进排卵关键过程。