超促排卵周期中巨噬细胞集落刺激因子的测定

目的　探讨巨噬细胞集落刺激因子 (M CSF)对卵泡发育、排卵、卵子受精及胚胎质量的影响。方法　用酶联免疫吸附试验方法测定 3 6例超促排卵患者的卵泡早期、卵泡中期和取卵日血清中M CSF水平 ,以及取卵日卵泡液中M CSF水平。结果　超促排卵周期中 ,血清M CSF水平呈逐渐升高趋势 ,取卵日达高峰。取卵日卵泡液M CSF水平 [(12 4 5 .7± 4 3 .6)kU/L]显著高于同日血清M CSF水平 [(983 .6± 2 9.6)kU/L]。在找到卵子、受精卵及直径≥ 16mm和容积≥ 2ml的卵泡中 ,其卵泡液M CSF水平分别为 (13 2 7.3± 4 2 .8)kU/L、(13 5 6.2± 3 4.7)kU/L和 (12 97.6± 3 3 .7)kU/L ;而未找到卵子、未受精及直径 <16mm和容积 <2ml卵泡的卵泡液M CSF水平分别为 (10 86.7± 2 8.3 )kU/L、(1175 .4± 3 7.3 )kU/L和 (10 3 8.4± 2 5 .9)kU/L ,3者间比较 ,差异均有极显著性 (P <0 .0 1,P <0 .0 5 ,P<0 .0 5 )。但是 ,胚胎分级 3～ 4分的卵泡液M CSF水平 [(12 71.8± 3 7.3 )kU/L]与胚胎分级 1～ 2分的卵泡液M CSF水平 [(13 12 .6± 5 1.2 )kU/L]比较 ,差异无显著性 (P >0 .0 5 )。结论　M CSF通过参与卵泡发育、成熟而影响卵子受精 ,但不影响卵裂及胚胎质量     

巨噬细胞集落刺激因子对卵泡颗粒黄体细胞性激素分泌的影响及其受体的测定

目的　了解人卵泡颗粒黄体细胞中巨噬细胞集落刺激因子 (M CSF)受体的表达 ,M CSF在体外对人卵泡颗粒黄体细胞产生雌、孕激素的影响。方法　采用免疫细胞化学染色法 ,测定 2 0例行卵母细胞浆内单精子注射治疗患者的卵泡颗粒黄体细胞的M CSF受体 ;采用酶免疫分析法 ,测定卵泡颗粒黄体细胞在M CSF(浓度分别为 0、10、2 5、5 0、10 0、2 5 0ng/ml)单独及与促卵泡激素 (FSH ,浓度75IU/ml)联合作用 72h后的上清液中雌二醇 (E2 )和孕酮 (P)的浓度。结果　约 80 %的颗粒黄体细胞膜上M CSF受体阳性。在无FSH存在时 ,M CSF空白对照颗粒黄体细胞培养上清液中E2 浓度为(2 185± 189)pmol/L ,P浓度为 (315 7± 4 0 1)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (2 789± 36 5 )～ (42 82± 318)pmol/L之间 ,P浓度在 (42 5 6± 5 95 )～ (7789± 82 8)nmol/L之间。在有FSH作用时 ,M CSF空白对照的E2 和P浓度分别为 (5 0 4 5± 4 86 )pmol/L和 (86 6 7± 92 3)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (6 5 6 7± 6 73)～ (8373± 935 )pmol/L之间 ,P浓度在 (10 999± 985 )～ (14 990±115 8)nmol/L之间。M CSF促颗粒黄体细胞E2 和P分泌的作用随浓度增加而增强 (P均 <0 0 5 ) ;M CSF与FSH共同作用下颗粒黄体     

巨噬细胞集落刺激因子在卵泡黄素化颗粒细胞中的表达

目的　了解巨噬细胞集落刺激因子 (M CSF)在人卵泡黄素化颗粒细胞中的表达 ,探讨促卵泡激素 (FSH)在体外对人卵泡黄素化颗粒细胞合成分泌M CSF的调节作用。方法　取接受卵母细胞单精子显微注射治疗的 2 0例妇女的卵泡黄素化颗粒细胞 ,采用免疫细胞化学染色法测定黄素化颗粒细胞中M CSF的表达 ,并将黄素化颗粒细胞单独培养及在不同浓度的FSH(2、5、15、2 5IU/ml)作用下培养 72h ,采用酶联免疫吸附法测定黄素化颗粒细胞培养液中的M CSF含量。结果　约 80 %的黄素化颗粒细胞胞浆有M CSF表达 ;黄素化颗粒细胞在培养 72h后 ,基础状态下M CSF分泌量为(5 9± 13)ng/L ,浓度为 2、5、15、2 5IU/ml的FSH作用下M CSF分泌量分别为 (16 5± 32 )、(2 10± 5 8)、(2 99± 6 6 )、(390± 78)ng/L ,在不同浓度的FSH作用下黄素化颗粒细胞培养液中M CSF的含量与基础状态下相比 ,差异均有极显著性 (P <0 0 1)。结论　卵泡黄素化颗粒细胞能合成分泌M CSF ;FSH能促进黄素化颗粒细胞M CSF的分泌 ,FSH对卵泡发育和成熟的调节 ,可能部分通过M CSF及其受体通道完成     

雌孕激素对体外卵泡黄素化颗粒细胞分泌巨噬细胞集落刺激因子的影响

目的　探讨雌、孕激素对卵泡黄素化颗粒细胞 (颗粒细胞 )分泌巨噬细胞集落刺激因子(M CSF)的调节作用。方法　抽取接受卵母细胞单精子显微注射 (ICSI)的 12例妇女的颗粒细胞 ,在不同浓度 ( 0、1× 10 - 7、1× 10 - 6 、1× 10 - 5 、1× 10 - 4、1× 10 - 3 mol L)的雌二醇或孕酮作用下培养 72h ,采用酶联免疫吸附法测定颗粒细胞培养液中的M CSF含量。结果　颗粒细胞在雌二醇或孕酮为 0mol L(即无雌二醇或孕酮作用———基础状态 )的情况下培养 72h后 ,分泌一定量的雌二醇 [( 1 0 2± 0 2 3 )×10 - 7mol L]、孕酮 [( 1 0 0± 0 12 )× 10 - 5 mol L]和少量的M CSF[( 4 7± 15)ng L] ;在 1× 10 - 6 ～ 1× 10 - 3mol L浓度的雌二醇作用下 ,M CSF分泌增加 ,其分泌随雌二醇浓度的增加而增加 ,较基础状态分别增加 2 3、4 5、6 9和 7 9倍 ;在≥ 1× 10 - 6 mol L各浓度的雌二醇作用下 ,M CSF分泌与基础状态比较 ,差异均有显著性 (P均 <0 0 5) ;在 1× 10 - 7mol L浓度的雌二醇作用下 ,M CSF分泌与基础状态比较 ,差异无显著性 (P >0 0 5) ;在 1× 10 - 7～ 1× 10 - 3 mol L浓度的孕酮作用下 ,M CSF分泌与基础状态比较 ,差异均无显著性 (P >0 0 5)。结论　雌二醇能促进颗粒细胞M CSF的分泌 ,孕酮对颗     

血管内皮生长因子在预测卵巢过度刺激综合征中的作用

目的探讨在体外受精-胚胎移植(IVF-ET)周期中血清及卵泡液中血管内皮生长因子(VEGF)水平的变化,及其在预测卵巢过度刺激综合征(OHSS)中的作用.方法收集42例行IVF-ET患者的血清和卵泡液标本,采用定量酶联免疫方法检测血清、卵泡液中VEGF水平,按是否发生OHSS分为OHSS组(10例)及对照组(32例),对两组临床资料、性激素和VEGF水平进行回顾性相关分析.结果 OHSS组血清VEGF水平在月经周期的各个时期均高于对照组,但差异无显著性(P>0.05).OHSS组卵泡液VEGF水平为(1 487.7±365.8) ng/L,较对照组的(1 025.8±474.7) ng/L明显升高;取卵时卵泡数目增多,OHSS组为(10.0±5.9)个,对照组为(6.1±2.3)个;基础黄体生成素(LH),人绒毛膜促性腺激素(hCG)、注射日雌二醇(E2)水平均比对照组高,差异有显著性(P<0.05).结论 OHSS组卵泡液VEGF水平明显高于对照组,提示VEGF可能参与了OHSS的发病;卵泡液VEGF水平可以作为预测OHSS发生的检测指标.     

移动抑制因子与体外受精-胚胎移植妊娠结局的相关性研究

目的:探讨人卵泡液中是否存在移动抑制因子(MIF)及其卵泡液和血液中的浓度对IVF-ET 妊娠结局的影响。方法:选择2004．08-2005．08行IVF-ET助孕的不孕妇女共37例,按妊娠结局分 为妊娠和非妊娠两组。收集取卵时的血液及卵泡液,定量酶联免疫法(ELISA)分别测定血及卵泡液 中MIF的浓度。并检测取卵前血中LH、P、E2值。结果:两组在获卵数、受精率、卵裂率、移植 胚胎数和激素水平无显著差异。妊娠组血清及卵泡液中MIF浓度分别为1．79±0．98 ng／ml和 22．97±10．62 ng／ml;非妊娠组分别为1．91±1．71 ng／ml和13．44±8．5ng/ml。两组卵泡液中 MIF浓度比较有显著差异(P<0．05);血清中浓度差异不明显(P>0．05)。卵泡液MIF浓度与优质胚 胎率呈正相关(r=0．3277,P<0．05),优质胚胎率在两组间差异显著(P<0．02)。结论:卵泡液中MIF可 能与卵泡发育及卵细胞质量有关,进而影响IVF-ET的结局。     

血管内皮生长因子在卵巢过度刺激综合征发病机理中作用的初步研究

目的通过对卵巢过度刺激综合征(OHSS)患者血管内皮生长因子(VEGF)水平和表达的测定,初步探讨VEGF在OHSS发病机理中的作用.方法采用酶联免疫吸附试验方法对14例中、重度OHSS患者(OHSS组)和同期13例非OHSS患者(对照组)取卵日卵泡液、人绒毛膜促性腺激素(hCG)注射日、胚胎移植(ET)日血清VEGF水平进行测定;采用免疫组织化学方法和逆转录聚合酶链反应技术检测取卵日卵巢黄素化颗粒细胞VEGF及其mRNA的表达.结果 OHSS组和对照组的卵泡液、hCG注射日血清VEGF水平分别为(1 257.2±648.0)与(1 745.1±802.4) ng/L,(250.1±109.5)与(196.7±81.7) ng/L,两组比较,差异无显著性(P>0.05);OHSS组ET日血清VEGF水平及注射hCG后血清VEGF水平变化程度,显著高于对照组[分别为(342.9±158.5)与(222.2±84.6) ng/L,(92.8±106.7)与(25.9±21.8) ng/L,P<0.05].OHSS组取卵日卵巢黄素化颗粒细胞VEGF mRNA的表达,显著高于对照组,VEGF/β肌动蛋白比值分别为1.50±0.60、0.96±0.56(P<0.05).两组取卵日卵巢黄素化颗粒细胞有较活跃的VEGF表达.结论 VEGF在OHSS的发病机理中可能起一定的作用.     

卵泡液中白血病抑制因子(LIF)和血清中E_2在早期胚胎发育中的作用

目的:阐述卵泡液中白血病抑制因子(LIF)和血清中E2在早期胚胎发育中的作用。方法:利用临床62例控制性超促排卵(COH)IVF-ET患者治疗过程中测定的血清E2数据和取卵时收集到的卵泡液,采用ELISA法测定卵泡液中LIF含量,分析LIF在取卵日不同E2组中的差异,并进一步分析其在卵母细胞、胚胎发育中的作用。结果:LIF在E2=1500-4000pg/ml组最高,在E2<1500pg/ml组中表达最低;在不同年龄组表达无差异。LIF<15pmol/L和≥15pmol/L组间受精率、可用胚胎率均存在显著性差异。LIF浓度与受精率、可用胚胎率成正相关。结论:受精率及早期胚胎的质量可能与卵泡液中LIF含量有关。卵泡液中LIF的浓度受周围血E2的影响;合适的E2浓度有利于卵泡液中LIF的提高。     

卵泡液中雌孕激素水平及卵泡大小与卵母细胞成熟度的关系

目的:探讨卵胞浆内单精子显微注射(ICSI)周期取卵日卵泡大小、卵泡液中雌孕激素水平和卵母细胞成熟度的相关性。方法:选择因男方因素在本中心接受ICSI治疗的51例不孕患者作为研究对象,收集取卵日卵泡液共140例,取卵前B超测量卵泡平均直径,采用电化学发光法(ECLI)检测卵泡液中雌二醇(E2)、孕酮(P)水平,分析取卵日卵泡大小、卵母细胞成熟度及卵泡液中雌孕激素水平三者间的相互关系。结果:卵泡平均直径与卵泡液E2水平呈正相关(r=0.724,P<0.05),与P水平亦呈正相关(r=0.798,P<0.05);按卵母细胞成熟度分为MⅡ组、MⅠ组、GV组和退化组,4组卵泡液中P水平无统计学差异(P>0.05),但退化组卵泡液中E2水平显著低于MⅡ组(P<0.05);所有样本按P/E2值分为1(<15)、2(15～25)、3(>25)组,2组卵母细胞成熟率最高,与1、3组的差异均有统计学意义(P<0.017),1组与3组的卵母细胞成熟率无统计学差异(P>0.017)。根据卵泡平均直径分组,直径>1.6cm为大卵泡组,1.6～1.2cm为中卵泡组,<1.2cm为小卵泡组,大卵泡组卵母细胞成熟率最高,与中、小卵泡组的差异有统计学意义(P<0.017),中卵泡组卵母细胞成熟率显著高于小卵泡组(P<0.017)。结论:卵泡大小与卵泡液雌孕激素水平呈正相关。卵泡液中P/E2在特定范围内时卵母细胞成熟率较高,其水平过高或过低均影响卵母细胞成熟;大卵泡组与中、小卵泡组在卵母细胞成熟率方面有显著统计学差异。     

绝经过渡期妇女血清抑制素水平的研究

目的　分析绝经过渡期妇女血清抑制素A(Inh A)、抑制素B(Inh B)水平的变化及其与其他生殖激素水平变化的时间关系。方法　测定 10例正常育龄妇女月经周期各期血清Inh A、Inh B水平 ;测定 40例绝经过渡期妇女月经周期第 3天的血清Inh B、促卵泡激素 (FSH)、促黄体生成素(LH)、雌二醇 (E2 )水平 ,经前 5～ 9d(经前期 )血清Inh A、孕酮 (P)水平 ;测定 10例绝经后妇女的血清Inh A、Inh B、FSH、E2 水平。分析各项激素水平变化之间的时间关系。结果　育龄妇女月经周期中Inh A、Inh B水平的变化曲线各不相同。绝经过渡期妇女中 ,黄体功能正常者占 48% ,经前期Inh A水平低于育龄妇女 ,分别为 ( 2 4 7± 13 0 )及 ( 42 9± 12 1)ng/L ,两者比较 ,差异有显著性 (P =0 0 17) ,Inh B水平改变差异无显著性 (P >0 0 5 ) ;黄体功能不足与无排卵者的Inh A水平进一步显著下降 ,分别为 ( 12 4± 10 2 )及 ( 5 3± 3 8)ng/L(P分别为 0 0 3 3及 <0 0 0 0 1) ,绝经后妇女则皆未检出。与育龄妇女比较 ,月经周期第 3天Inh B水平的下降仅在无排卵与绝经后妇女差异有极显著性 (P =0 0 0 1)。月经周期第 3天 ,FSH≥ 10IU/L者Inh B水平显著低于FSH <10IU/L者 ,分别为 ( 16 2± 4 0 )及 ( 62 0±43 8)ng/L(P <0 0 0     

人宫颈粘液葡萄糖、果糖变化规律的研究

本文对月经周期规律的１５例育龄妇女２０个月经周期宫颈粘液及血清葡萄糖、果糖水平进行测定，同时对血清促黄体激素（ＬＨ）、促卵泡激素（ＦＳＨ）、雌二醇（Ｅ２）和孕酮（Ｐ）放免测定；阴道Ｂ超、宫颈粘液Ｉｎｓｌｅｒ评分；尿ＬＨ酶联免疫测定和基础体温（ＢＢＴ）测定，综合评价预测和确定排卵日。２０个周期均为有排卵周期。结果：宫颈粘液葡萄糖、果糖水平均表现为卵泡期稍高，排卵前最低，排卵后逐渐升高，黄体中期达高峰；卵泡期果糖水平与Ｅ２呈负相关（ｒ＝－０．７３，Ｐ＜０．０１）；黄体期葡萄糖、果糖水平均与Ｐ呈正相关（ｒ＝０．９９，Ｐ＜０．０１；ｒ＝０．９８，Ｐ＜０．０１）；血葡萄糖、果糖水平无周期性变化且与宫颈粘液葡萄糖、果糖水平无明显相关性（ｒ＝０．２３，Ｐ＞０．０５）。结论：宫颈粘液葡萄糖、果糖水平变化具有明显规律性，可能与雌孕激素调节有关。     

超促排卵对不孕女性子宫内膜蠕动波的影响

目的:研究不孕女性自然周期与控制性超促排卵(COH)周期子宫内膜蠕动波的特点。方法:64名排卵正常的不孕女性分别于自然周期LH峰日、排卵日、排卵后2 d和COH周期hCG注射后1 d、采卵日、采卵后2 d阴道超声监测子宫内膜蠕动波,且同时测定血清雌、孕激素水平。结果:自然周期子宫内膜蠕动波频率是COH周期的1.31倍;COH周期与自然周期各个观测日的子宫内膜蠕动波类型分布不同;子宫内膜蠕动波频率与生理水平血清雌二醇(E2)呈正相关,与孕酮(P)呈负相关,与超生理剂量的雌、孕激素无相关性。结论:COH治疗显著地改变了子宫内膜蠕动波的自然运动模式,在胚胎移植前仍表现为较强烈的子宫内膜运动。     

Altered follicular development in clomiphene citrate versus human menopausal gonadotropin-stimulated cycles for in vitro fertilization

The pattern of periovulatory and luteal phase serum estradiol (E2) and progesterone (P) as well as follicular fluid (FF) E2, P, androgen, gonadotropin, and prolactin concentrations of eight women undergoing clomiphene citrate (CC)/human chorionic gonadotropin (hCG) stimulation and eight women undergoing human menopausal gonadotropin (hMG)/hCG stimulation of follicular development for the purpose of in vitro fertilization were compared. Ovulation was induced with either a 5-day course of CC (100 mg/day beginning on day 5 of the cycle) or an individualized hMG regimen, and laparoscopy was performed 36 hours after hCG administration. The length of the luteal phase was significantly longer (P less than 0.05) in the CC-treated group as compared with the hMG-treated group. The pattern of serum E2 levels differed significantly (P less than 0.01) in that E2 levels were lower in the early and midluteal phase in CC-stimulated cycles; in addition, a delayed second E2 peak was observed in the late luteal phase in these women. Serum P levels, however, were lower in the hMG-stimulated group. Analysis of FF hormone concentrations revealed significantly (P less than 0.05) higher concentrations of E2 and androsterone in the FF of hMG-treated patients. It is concluded that follicular development in CC-stimulated cycles differs markedly from that in hMG-stimulated cycles. These differences may reflect either an altered follicular maturational process or may represent a direct inhibitory effect of CC on follicular steroidogenesis.     

Expression of Integrin Fraction and Adhesion Molecules on Human Granulosa Cells and Its Relation with Oocyte Maturity and Follicular Steroidogenesis

PURPOSE: To study the correlation between the expression of integrin fractions and adhesion molecules on granulosa cells (GC) and follicular development. METHODS: GC and follicular fluid (FF) were obtained at oocyte retrieval for ICSI. Expression of adhesion molecules on GC was studied by flow cytometry. Statistics were evaluated using the Student t test and simple linear regression analysis. RESULTS: alpha5 integrin fraction was significantly (p < 0.01) higher, while alpha6 fraction and CD9 were significantly (p < 0.01 and p < 0.001, respectively) lower in GC from FF with metaphase II oocytes. A direct significant correlation was observed between FF progesterone and the alpha5 expression on GC (r = 0.54). In contrast, an inverse significant correlation was observed between FF progesterone level and the expression of alpha6 and CD9 (r = -0.40 and -0.41, respectively). CONCLUSIONS: The expression pattern for integrin fractions and adhesion molecules may be of predictive value in assessing the state of differentiation of the human follicle.     

血清抑制素含量在月经周期中的变化及其与卵泡刺激素的相关性

目的了解育龄妇女在月经周期中血清抑制素(INH)含量的变化规律及其与卵泡刺激素(FSH)、黄 体生成素(LH)、雌二醇(E2)及孕酮(P)的相关性。方法建立一种改良的INH放射免疫测定法(RIA),对育龄妇 女正常月经周期中及绝经后妇女血清INH含量的变化进行监测;同时测定血清FSH、LH、E     

Renin-like activity in ovarian follicular fluid

Angiogenesis is a feature of ovarian follicle and corpus luteum development. Ovarian homogenates and follicular fluid (FF) contain factors that stimulate new vessel formation. Because the renin-angiotensin system has been shown to facilitate angiogenesis, renin activity was measured in FF and plasma from 20 normal, ovulatory women who were undergoing in vitro fertilization and were therefore stimulated with human menopausal gonadotropin/human chorionic gonadotropin. Serum estradiol (E2) and FF E2 and progesterone (P) were also determined and correlated with FF and plasma renin activity. FF renin activity was significantly higher than plasma renin activity (55.8 +/- 7.9 versus 3.8 +/- 0.7 ng of angiotensin I/ml/hour, P less than 0.001). Positive correlations were found between FF E2 and renin activity (r = 0.53, P less than 0.05). There was also a correlation between plasma renin activity and serum E2 (r = 0.69, P less than 0.05). No correlation was present between FF P and renin activity. The high renin-like activity present in FF after stimulation with gonadotropins could be involved in the mechanism of angiogenesis and may play an important role in events related to reproductive processes.     

High progesterone/estradiol ratio in follicular fluid at oocyte aspiration for in vitro fertilization as a predictor of possible pregnancy

Eighteen women undergoing in vitro fertilization (IVF) procedures were studied. All had optimal (900 to 1600 pg/ml) peak serum estradiol (E2) response to the same stimulation regimen with clomiphene citrate and menotropins; fertilization rate was above 64%; and two to four embryos in two to eight cell stages were replaced in each patient. All were considered to have optimal chances for conception. The authors compared progesterone (P), E2, and P/E2 ratio in serum and follicular fluid (FF) at the time of oocyte aspiration in eight patients who conceived (group I) and ten who did not (group II). Mean serum P and E2 levels and serum P/E2 ratio were not significantly different between the groups. In contrast, mean FF P concentrations (ng/ml) were significantly (P less than 0.05) higher in group I (9721 versus 5385), as was FF P/E2 ratio (19.0 versus 11.8; P less than 0.02). There was no significant difference in mean FF E2 concentrations between the groups. These data indicate that in IVF cycles with optimal serum E2 response to the stimulation protocol, FF P and P/E2 ratio at the time of oocyte aspiration may be predictive of subsequent implantation and pregnancy.     

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. I. Sex steroids

The steroid content of 72 follicular fluids (FF) obtained from 42 women subjected to ovulation induction with the luteinizing hormone-releasing hormone analogue D-Trp6 and human menopausal gonadotropin was studied in terms of the evolution of in vitro fertilization (IVF) and embryo transfer (ET) results. The FF were classified into several categories based on oocyte evolution. Individual values of FF estrone and estradiol (E2), as well as androstenedione and testosterone could not be correlated with ET outcome. However, FF progesterone (P) levels for follicles leading to pregnancy were significantly lower when compared with those in the other categories. The correlation between the E2/P ratio and E2 permitted the definition of a band wherein IVF-ET outcome was successful and enabled the characterization of different functional follicular maturational states.