多囊卵巢综合征卵泡发育障碍及滋阴补阳中药序贯干预的机理研究
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摘要
目的：通过研究滋阴补阳中药序贯对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞(GC)分泌功能和细胞因子基因表达的调控,以及滋阴补阳中药序贯对PCOS不孕患者促排卵周期、体外受精-胚胎移植(IVF-ET)中血清、卵泡液中干细胞因子(SCF)水平的影响及其与优势卵泡、卵细胞质量的关系,探讨滋阴补阳中药序贯对PCOS干预的作用机理。
方法：1.实验研究：将造模成功的PCOS大鼠,按实验要求分离与培养卵巢颗粒细胞,并制备空白血清、滋阴方、补阳方含药血清,观察滋阴方、补阳方对PCOS大鼠卵巢颗粒细胞分泌E2、P功能的影响,以及对干细胞因子(SCF)、雄激素受体(AR)、类固醇合成急性调控蛋白(StAR)基因表达调控的影响。2.临床研究：选取PCOS患者60例,其中经典PCOS30例,无高雄PCOS30例；并选取非PCOS患者30例作为正常组对照,分析基础状态下SCF与PCOS发病机制的相关性。另选取62例行促排卵治疗的PCOS患者随机分为2组,治疗组(滋阴补阳中药序贯／克罗米芬(CC)／果纳芬(r-FSH),30个周期)和对照组(CC/r-FSH,32个周期)。观察两组患者HCG注射天数、子宫内膜厚度、优势卵泡个数、HCG日血清E2、LH、SCF水平、比较两组在排卵率、妊娠率、卵巢过度刺激综合征(OHSS)以及卵泡未破裂黄素化综合征(LUFS)等方面的效果。40例将行IVF-ET的PCOS患者随机分为2组,治疗组(滋阴补阳中药序贯联合西药组)和对照组(西药组),每组各20例。观察两组患者HCG日血清E2、LH、P水平、取卵日卵泡液中SCF水平以及取卵个数、受精率、卵裂率、优质胚胎率、妊娠率。
结果：1.造模后,PCOS大鼠卵巢大体解剖以及HE染色后卵巢光镜均有明显不同。2.卵泡期中,滋阴方10%、20%组均可有效提高PCOS大鼠颗粒细胞上清液中的E2、SCF含量；黄体期中,补阳方10%组刺激黄素化颗粒细胞分泌P的作用最强。当补阳方浓度继续增大,其促进P分泌的作用并没有增强。3.模型组PCOS大鼠颗粒细胞分泌E2、P水平显著降低。10%、20%滋阴方、20%补阳方均可有效促进PCOS大鼠卵巢颗粒细胞分泌E2、P,滋阴方的浓度与颗粒细胞分泌E2、P成一定的量效关系。4.模型组大鼠卵巢GC中ARmRNA表达显著升高,SCF、StARmRNA表达显著降低。10%、20%滋阴方组、20%补阳方组可有效下调ARmRNA表达,上调SCF、StARmRNA表达。5.无高雄PCOS AFC明显多于经典PCOS组、正常组。无高雄PCOS组与正常组T水平显著低于经典PCOS组。无高雄PCOS组血清SCF水平明显升高。无高雄PCOS组SCF水平与AFC呈正相关。6.PCOS患者促排卵治疗后,试验组治愈率36.7%,总有效率86.7%；对照组治愈率12.5%,总有效率87.5%,两组治愈率有显著差异。试验组HCG日子宫内膜厚度、HCG日≥14mm卵泡数、HCG日≥18mm卵泡数均有显著差异。试验组HCG日血清E2、SCF明显高于对照组。7. IVF-ET周期治疗组取卵日卵泡液中SCF水平、受精率、优质胚胎率明显高于对照组。妊娠组取卵日卵泡液SCF水平显著高于未妊娠组。
结论：1.SCF与PCOS卵泡发育障碍的发病机制相关。2.滋阴、补阳方可明显改善PCOS大鼠卵巢颗粒细胞分泌功能,下调AR基因表达,上调SCF、StAR基因表达。3.滋阴补阳中药序贯联合西药促排卵可以增加子宫内膜厚度及HCG日E2、SCF水平,提高妊娠率。4.对PCOS患者IVF-ET中进行滋阴补阳中药序贯干预,可改善卵巢内微环境,提高卵泡液内SCF水平,从而提高受精率、优质胚胎率、妊娠率。
Objective:Through sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention to study the mechanism of the therapy controlling secretion regulation function and expression of cytokine genes in ovarian granulosa cells in PCOS rats.Furthermore, monitor stem cell factor(SCF) level in serum,follicular fluid by ovulation induction cycle or in vitro fertilization and embryo transfer (IVF-ET) cycle on infertile patients with PCOS,besides its relationship with the influence of dominant follicle and oocyte quality.
Methods:1. Experiment Study:According to the experimental requirements establish PCOS rats model,then, isolate and culture ovarian granulosa cells in different periods.Prepare three different medicated serum namely blank serum, nourishing Yin serum, supplementing Yang serum, to observe their effects on PCOS rats'ovarian granulosa cell secretion function of E2, P,and the influence of stem cell factor, androgen receptor, steroidogenic acute regulatory protein gene expression.2. Clinical research:30cases of classic PCOS,30cases PCOS without hyperandrogenism,30patients without PCOS were selected to analysis correlation of SCF and PCOS pathogenesis in basic state. Another62cases PCOS patients are randomly divided into the treatment group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine/clomiphene citrate/Gonal-F,30cycles) and control group (clomiphene citrate/,Gonal-F,32cycles).Two groups were observed with treatment days, endometrial thickness, the dominant follicles number,serum E2, LH, SCF levels in the HCG injection day and compared ovulation rate, pregnancy rate,ovarian hyperstimulation syndrome (OHSS) and luteinized unruptured follicle syndrome (LUFS) etcetera.40cases of PCOS patients of IVF-ET were randomly divided into into2groups, experimental group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine group) and control group (Western medicine group), each group of20cases. Observation serum E2, LH, P levels in HCG injection day,SCF levels in follicular fluid and the difference of retrieved oocyte, fertilization rate, cleavage rate, high quality embryo rate and pregnancy rate.
Results:1. PCOS rat ovarian anatomy and HE staining light microscope ovarian are obviously different.2. In follicular phase, nourishing Yin Decoction10%,20%groups can improve PCOS rats'E2, SCF content in granulosa cell,similarly in luteal phase, supplementing Yang Decoction20%group displayed the strongest function of stimulating granulosa cells to secret P,but no enhancement when concentration increasing. No statistical significance among the groups of SCF level.3. There were significantly reduced the granulosa cells secretion of E2and P in model group.Both nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can promote PCOS rats' ovarian granulosa cells to secret E2, P.And that nourishing Yin Decoction concentration has a certain dose-effect relationship with granulosa cell secretion level of E2, P.4. Expression of AR mRNA in model group was significantly increased, but SCF, StAR mRNA expression decreased obviously.while,nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can effectively reduce the granulosa cell AR mRNA expression, increase SCF, StAR mRNA expression.5. The without hyperandrogenism PCOS group AFC was significantly more than the classical PCOS group, normal group,and had a significantly elevated SCF levels,show a positive correlation with AFC. But,its T level,besides the normal group,was significantly lower than the classical PCOS group.6. The treatment group's cure rate was36.7%and total efficiency of86.7%,meanwhile,control group, the cure rate was12.5%, the total efficiency of87.5%, two group had significant difference in the cure rate, endometrial thickness,above14mm and18mm follicles number in HCG injection day. Treatment group's serum E2, SCF level in HCG injection day were significantly higher than those in control group. The test group was higher than that of the control group pregnancy.7. In IVF-ET cycles, the follicular fluid levels of SCF in retrieved oocyte day, fertilization rate, high quality embryo rate was significantly higher in treatment group han that of control group. In either group,pregnant patients'follicular fluid SCF levels were significantly higher than those not pregnant in retrieved oocyte day.
Conclusion1. SCF relates to the pathogenesis of PCOS follicles developmental disorders.2. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention can significantly improve the PCOS rats' ovarian granulosa cells secretion function, down-regulate AR gene expression, up-regulate SCF, StAR gene expression.3. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine to induce ovulation can increase the endometrial thickness, serum E2, SCF levels in HCG injection day, improve the pregnancy rate.4. IVF-ET patients with PCOS,intervent by sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine, can improve the ovarian microenvironment, improve the SCF level in follicular fluid, so as to improve the fertilization rate, high quality embryo rate, pregnancy rate.
方法：1.实验研究：将造模成功的PCOS大鼠,按实验要求分离与培养卵巢颗粒细胞,并制备空白血清、滋阴方、补阳方含药血清,观察滋阴方、补阳方对PCOS大鼠卵巢颗粒细胞分泌E2、P功能的影响,以及对干细胞因子(SCF)、雄激素受体(AR)、类固醇合成急性调控蛋白(StAR)基因表达调控的影响。2.临床研究：选取PCOS患者60例,其中经典PCOS30例,无高雄PCOS30例；并选取非PCOS患者30例作为正常组对照,分析基础状态下SCF与PCOS发病机制的相关性。另选取62例行促排卵治疗的PCOS患者随机分为2组,治疗组(滋阴补阳中药序贯／克罗米芬(CC)／果纳芬(r-FSH),30个周期)和对照组(CC/r-FSH,32个周期)。观察两组患者HCG注射天数、子宫内膜厚度、优势卵泡个数、HCG日血清E2、LH、SCF水平、比较两组在排卵率、妊娠率、卵巢过度刺激综合征(OHSS)以及卵泡未破裂黄素化综合征(LUFS)等方面的效果。40例将行IVF-ET的PCOS患者随机分为2组,治疗组(滋阴补阳中药序贯联合西药组)和对照组(西药组),每组各20例。观察两组患者HCG日血清E2、LH、P水平、取卵日卵泡液中SCF水平以及取卵个数、受精率、卵裂率、优质胚胎率、妊娠率。
结果：1.造模后,PCOS大鼠卵巢大体解剖以及HE染色后卵巢光镜均有明显不同。2.卵泡期中,滋阴方10%、20%组均可有效提高PCOS大鼠颗粒细胞上清液中的E2、SCF含量；黄体期中,补阳方10%组刺激黄素化颗粒细胞分泌P的作用最强。当补阳方浓度继续增大,其促进P分泌的作用并没有增强。3.模型组PCOS大鼠颗粒细胞分泌E2、P水平显著降低。10%、20%滋阴方、20%补阳方均可有效促进PCOS大鼠卵巢颗粒细胞分泌E2、P,滋阴方的浓度与颗粒细胞分泌E2、P成一定的量效关系。4.模型组大鼠卵巢GC中ARmRNA表达显著升高,SCF、StARmRNA表达显著降低。10%、20%滋阴方组、20%补阳方组可有效下调ARmRNA表达,上调SCF、StARmRNA表达。5.无高雄PCOS AFC明显多于经典PCOS组、正常组。无高雄PCOS组与正常组T水平显著低于经典PCOS组。无高雄PCOS组血清SCF水平明显升高。无高雄PCOS组SCF水平与AFC呈正相关。6.PCOS患者促排卵治疗后,试验组治愈率36.7%,总有效率86.7%；对照组治愈率12.5%,总有效率87.5%,两组治愈率有显著差异。试验组HCG日子宫内膜厚度、HCG日≥14mm卵泡数、HCG日≥18mm卵泡数均有显著差异。试验组HCG日血清E2、SCF明显高于对照组。7. IVF-ET周期治疗组取卵日卵泡液中SCF水平、受精率、优质胚胎率明显高于对照组。妊娠组取卵日卵泡液SCF水平显著高于未妊娠组。
结论：1.SCF与PCOS卵泡发育障碍的发病机制相关。2.滋阴、补阳方可明显改善PCOS大鼠卵巢颗粒细胞分泌功能,下调AR基因表达,上调SCF、StAR基因表达。3.滋阴补阳中药序贯联合西药促排卵可以增加子宫内膜厚度及HCG日E2、SCF水平,提高妊娠率。4.对PCOS患者IVF-ET中进行滋阴补阳中药序贯干预,可改善卵巢内微环境,提高卵泡液内SCF水平,从而提高受精率、优质胚胎率、妊娠率。
Objective:Through sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention to study the mechanism of the therapy controlling secretion regulation function and expression of cytokine genes in ovarian granulosa cells in PCOS rats.Furthermore, monitor stem cell factor(SCF) level in serum,follicular fluid by ovulation induction cycle or in vitro fertilization and embryo transfer (IVF-ET) cycle on infertile patients with PCOS,besides its relationship with the influence of dominant follicle and oocyte quality.
Methods:1. Experiment Study:According to the experimental requirements establish PCOS rats model,then, isolate and culture ovarian granulosa cells in different periods.Prepare three different medicated serum namely blank serum, nourishing Yin serum, supplementing Yang serum, to observe their effects on PCOS rats'ovarian granulosa cell secretion function of E2, P,and the influence of stem cell factor, androgen receptor, steroidogenic acute regulatory protein gene expression.2. Clinical research:30cases of classic PCOS,30cases PCOS without hyperandrogenism,30patients without PCOS were selected to analysis correlation of SCF and PCOS pathogenesis in basic state. Another62cases PCOS patients are randomly divided into the treatment group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine/clomiphene citrate/Gonal-F,30cycles) and control group (clomiphene citrate/,Gonal-F,32cycles).Two groups were observed with treatment days, endometrial thickness, the dominant follicles number,serum E2, LH, SCF levels in the HCG injection day and compared ovulation rate, pregnancy rate,ovarian hyperstimulation syndrome (OHSS) and luteinized unruptured follicle syndrome (LUFS) etcetera.40cases of PCOS patients of IVF-ET were randomly divided into into2groups, experimental group (sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine group) and control group (Western medicine group), each group of20cases. Observation serum E2, LH, P levels in HCG injection day,SCF levels in follicular fluid and the difference of retrieved oocyte, fertilization rate, cleavage rate, high quality embryo rate and pregnancy rate.
Results:1. PCOS rat ovarian anatomy and HE staining light microscope ovarian are obviously different.2. In follicular phase, nourishing Yin Decoction10%,20%groups can improve PCOS rats'E2, SCF content in granulosa cell,similarly in luteal phase, supplementing Yang Decoction20%group displayed the strongest function of stimulating granulosa cells to secret P,but no enhancement when concentration increasing. No statistical significance among the groups of SCF level.3. There were significantly reduced the granulosa cells secretion of E2and P in model group.Both nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can promote PCOS rats' ovarian granulosa cells to secret E2, P.And that nourishing Yin Decoction concentration has a certain dose-effect relationship with granulosa cell secretion level of E2, P.4. Expression of AR mRNA in model group was significantly increased, but SCF, StAR mRNA expression decreased obviously.while,nourishing Yin Decoction10%,20%groups and supplementing Yang Decoction20%group can effectively reduce the granulosa cell AR mRNA expression, increase SCF, StAR mRNA expression.5. The without hyperandrogenism PCOS group AFC was significantly more than the classical PCOS group, normal group,and had a significantly elevated SCF levels,show a positive correlation with AFC. But,its T level,besides the normal group,was significantly lower than the classical PCOS group.6. The treatment group's cure rate was36.7%and total efficiency of86.7%,meanwhile,control group, the cure rate was12.5%, the total efficiency of87.5%, two group had significant difference in the cure rate, endometrial thickness,above14mm and18mm follicles number in HCG injection day. Treatment group's serum E2, SCF level in HCG injection day were significantly higher than those in control group. The test group was higher than that of the control group pregnancy.7. In IVF-ET cycles, the follicular fluid levels of SCF in retrieved oocyte day, fertilization rate, high quality embryo rate was significantly higher in treatment group han that of control group. In either group,pregnant patients'follicular fluid SCF levels were significantly higher than those not pregnant in retrieved oocyte day.
Conclusion1. SCF relates to the pathogenesis of PCOS follicles developmental disorders.2. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine intervention can significantly improve the PCOS rats' ovarian granulosa cells secretion function, down-regulate AR gene expression, up-regulate SCF, StAR gene expression.3. Sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine with western medicine to induce ovulation can increase the endometrial thickness, serum E2, SCF levels in HCG injection day, improve the pregnancy rate.4. IVF-ET patients with PCOS,intervent by sequential therapy of nourishing Yin and supplementing Yang of traditional Chinese medicine, can improve the ovarian microenvironment, improve the SCF level in follicular fluid, so as to improve the fertilization rate, high quality embryo rate, pregnancy rate.
引文
[1]Polycystic ovary syndrome writing committee.American Association of Clinical Endocrinologists position statement on metabolic and cardio-vascular consequences of polycystic ovary syndrome. Endocr Pract,2005,11:126-134.
[2]Kahsar-MillerMD, Nixon C, Boots LR,et a.1 Prevalence of polycystlc ovary syndrome (PCOS) in first-degree relatives of patients with PCOS.FertilSteri,l 2001,75(1):53-58.
[3]Cooper HE,Spellacy WN,Prem KA,et al.Hereditary factors in the stein-leventhal syndrome.Am J Obstet Gynecol,1968,10:371-387.
[4]曹云霞,庄广伦.多囊卵巢综合征的染色体分析[J].中国优生与遗传杂志,2001,9(4)：37-38.
[5]Norman RJ, Dewailly D, Legro RS, et a.1 Polycystic ovary syndrome.Lancet,2007,370(9588); 685-697.
[6]Diamanti-Kandarakis E, Piperi C. Genetics of polycystic ovary syndrome:searching for the way out of the labyrinth [J]. Hum ReprodUpdate,2005,11(6):631-643.
[7]LegroRS, Strauss JF. Molecular progress in infertility:polycystic ovary syndrome [J]. FertilSteri,l 2002, 78(3):569-576.
[8]JakubowskiL.Genetic aspects ofpolycystic ovary syndrome.Endokry-nolPo,l 2005,56(3):285-293.
[9]李琳PCOS与青春期胰岛素抵抗.实用妇产科杂志,2005,21(9)：522-524.
[10]胡燕宁.少年多囊卵巢综合征与其父母代谢综合征的相关性研究[J].现代中西医结合杂志,2008,17(3)：335-337.
[11]KumarA,WoodsKS, BartolucciA, et a.1 Prevalence of adrenal androgen excess in patientswith the polycystic ovary syndrome(PCOS)[J].Clin Endocrino,l 2005,62(6):644-649.
[12]Gonzalez F,ChangL,HorabT,et al.Evidence for heterogeneous etiologies of adrenal dysfunction in polycystic ovary syndrome[J]. Fertil Steril,1996,66(3):354-361.
[13]Poretsky L, Piper B. Insulin resistance, hypersecretion of LH, and adual defect hypothesis for the pathogenesis of polycystic ovary syndrome[J]. Obstet Gynecol,1994,84(4):613-621.
[14]王蔼明,李美芝.多囊卵巢综合征患者血雄激素与胰岛素、黄体生成素水平关系的探讨[J].生殖医学杂志,2004,13(1)：47-49.
[15]Ludwig AK, Weiss JM, Tauchert S, et al. Influence of hypo-and hyperglycaemia on plasma leptin concentrations in healthy women and in women with polycystic ovary syndrome [J]. Hum Reprod,2007, 22(6):1555-1556.
[16]Fedorcsak P, Raki M, Storeng R. Characterization and depletion of leukocytes from cells isolated from the pre-ovulatory ovarian follicle [J]. Hum Reprod,2007,22(4):989-994.
[17]Stubbs SA, Stark J, Dilworth SM, et al. Abnormal preantral folliculogenesis in polycystic ovaries is associated with increased granulosa cell division [J]. J Clin Endocrinol Metab,2007,92(11):4418-4426.
[18]Webber LJ, Stubbs S, Stark J, et al. Prolonged survival in culture of preantral follicles form polycystic ovaries [J]. J Clin EndocrinolMetab,2007,92(5):1975-1978.
[19]GuthrieHD, GarrettWM, CooperBS. Follicle-stimula-ting hormone and insulin-like growth factor-Ⅰ attenuate apoptosis in cultured porcine granulosa cells [J].Biol Reprod,1998,58(2):390-396
[20]Coffey S,Mason H.The effect of polycystic ovary syndrome on health-related quality of life[J].Gynecol Endocrinol,2003,17(5):379-386.
[21]Elsenbruch S,Hahn S,Kowalsky D,et al.Quality of life,psychosocial well-being,and sexual satisfaction in women with polycystic ovary syndrome[J].J Clin Endocrinol Metab,2003,88(12):5801-5807.
[22]王玉蓉,曹云霞,丛林,等.多囊卵巢综合征患者的调查及心理分析[J].安徽医科大学学报,2005,40(3)：277-279.
[23]Eggerd S,Kirchengast S.The polycystic ovary syndrome a medical condition but also an important psychosocial problem[J].Coll Antropol,2001,25(2):673-685.
[24]Solomon CG.The epidemiology of polycystic ovary syndrome.Prevalence and associated disease risks.EndocrinolMetab Clin North Am,1999,28(2):247.
[25]Dumesic DA,Abbott DH.Implications of polycystic ovary syndrome on oocyte development[J].Semin Reprod Med,2008,26(1):53-61.
[26]Steckler T,Wang J,Bartol FF,et al.Fetal programming:prenatal testosterone treatment causes intrauterine growth retardation,reduces ovarian reserve and increases ovarian follicular recruitment[J]Endocrinology, 2005,146(7):3185-3193.
[27]Gougeon A.Regulation of ovarian follicular development in primates:facts and hypothesis[J].Endocr Rev,1996,17(2):121-155.
[28]kJakimiuk AJ,Weitsman SR,Brzechffa PR,et al.Aromatase mRNA expression in individual follicles from polycystic ovaries[J].MolHum Reprod,1998,4(1):1-8.
[29]D.S.Guzick,Polycystic ovary syndrome[J].Obstet Gynecol,2004,103:181-193.
[30]De Leo VDE, Marca ALA.Petiaglia F. Insulin-lowering agents in the management of polycystic ovary syndrome[J].Endocrine REV,2003,24(5):633-667.
[31]Kurzawa R,Ciepiela P,Baczkowski T, et al.Comparison of embryological and clinical outcome in GnRH antagonist vs.GnRH agonist protocols for in vitro fertilization in PCOS non—besepatients.Aprospective randomized study[J].J Assist Reprod Genet,2008,25(8):365-374.
[32]Vendola KA, Zhou J, Adesanya OO et al.Androgen stimulate early stages of follicular growth in the primate ovary.J Clin Invest,1998,101 (12):2622.
[33]乐杰,谢幸,林仲秋,等.妇产科学[M].第7版.北京：人民卫生出版社,2008：315.
[34]Greisen S,Ledet T,Ovesen P.Lack of association between polycystic ovary syndrome and embryonic aneuploidy[J].Hum Reprod,2001,16(10):2061-2065.
[35]Franks S,Stark J,Hardy K.Follicle dynamics and anovulation in polycystic ovary syndrome[J].Hum Reprod Update,2008,14(4):367-378.
[36]Picton HM,Harris SE,Muruvi W,et al.The in vitro growth and maturation of follicles[J] Reproduction, 2008,136 (6):703-715.
[37]Hillier SG.Current concepts of the roles of follicle stimulating hormone and luteinizing hormone in folliculogenesis[J].Hum Reprod,1994,9(2):188-191.
[38]Balen AH,ConwayGS,HomburgR,LegroRS. Polycystic Ovary Syndrome. A Guide to Clinical Management[J].London,UK:Taylor&Francis,2005,47-67.
[39]Kezele PR,Nilsson EE,Skinner MK.Insulin but not insulin-like growth factor-1 promotes the primordial to primary follicle transition[J].Mol Cell Endocrinol,2002,192(1-2):37-43.
[40]Balen AH,Conway GS,Homburg R,et al.Polycystic Ovary Syndrome-A Guide to Clinical Management [M].London:Taylor and Francis,2005:47-67.
[41]Zhang Y,Proenca R,Maffei M,et al. Positional cloning of the mouse obese gene and its human homologue [J]. Nature,1994,372(6505):425-432.
[42]Erturk E, Kurn N, Savci V et all Serum lep tin levels correlate with pbesity parameters but not with hyperin sulinism in women with polycystic ovary syndromelFertil Steril,2004,82 (5):1364.
[43]Li MG,Ding GL,Chen XJ,et al.Association of serum and follicular fluid leptin concentrations with granulosa cell phosphorylated signal transducer and activator of transcription 3 expression in fertile patients with polycystic ovarian syndrome[J].J Clin Endocrinol Metab,2007,92(12):4771-4776.
[44]Agarwal SK,Vogel K,Weitsman SR,et al.Leptin antagonizes the insulin-like growth factor-1 augmentation of steroidogenesis in granulosa and theca cells of the human ovary[J].J Clin EndocrinolMetab,1999,84 (3):1072-1076.
[45]Magoffin DA. Ovarian enzyme activities in women with Polycystic ovary syndrome. FertilSteril, 2006,86(7):9-11.
[46]NelsonVL,QinKnKN,RosenfieldRL,etal.The biochemical basis for increased testosterone production in Theca cells Propagated from patients with polycystic ovary syndrome[J],ClinEndocrinol Metab,2001, 86(12):5925-33.
[47]Stubbs SA, Hardy K, Da Silva-Buttkus P, et al. Anti-Mullerian hormone protein expression is reduced during the initial stages of follicle development in human polycystic ovaries. J Clin Endocrinol Metab, 2005,90(10):5536-43.
[48]Ehrmann DA,Liljenquist DR,Kasza K,et al.Prevalence and predictors of the metabolic syndrome in women with polycystic ovary syndrome[J].J Clin Endocrinol Metab.2006,91:48.
[49]林琳,刘复权,张晓莉,等Diane-35对多囊卵巢综合征患者肾素-血管紧张素系统及促排卵的影响[J].河北医科大学学报,2005,26(5)：370-372.
[50]Khattab S,Mohsen IA,Foutouh IA,et al.Metformin reduces abortion in pregnant women with polycystic ovary syndrome[J].GynecolEndcrinol,2006,22(12):680.
[51]赵军招,叶碧绿,林金菊,等.二甲双胍在多囊卵巢综合征促排卵治疗中的作用[J].中华妇产科杂志,2003,38(9)：545-548.
[52]The Rotterdam ESHRE/ASRM-Sponsored PCOSWorkshop Group.Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome[J]. Fertil Steri,l 2004,81(1): 19-25.
[53]Thessaloniki ESHRE/ASRM-Sponsored PCOS ConsensusWorkshop Group.Consensus on infertility treatment related to polycystic ovary syndrome[J]. Fertil Steri,l 2008,89(3):505-522.
[54]HolzerH,Casper R,Tulandi T.A new era in ovulation Induction[J].Ferti-lS teri,l 2006,85(2):277-284.
[55]Rossing MAIClomifene is used in women with Polycystic ovary syndrome[J].NEngl J Med,1994,331: 335-339.
[56]乔杰.多囊卵巢综合征的促排卵治疗[J].北京医科大学学报.1997,29(2)：190-192.
[57]张娜,等.氯米芬联合促卵泡生长激素低量缓增方案应用于多囊卵巢综合征效果观察[J].郑州大学学报(医学版).2007,42(4)：682-684.
[58]Tiboni G M.Aromatase inhibitors and teratogenesis[J].Fertility and Sterility,2004,81(4):1158-1159.
[59]Robert F,Mohamed F,Mitwally M,et al.Aromatase inhibitors for ovulation induction[J].J Clin Endocrinol Metabol,2005,91(3):760-761.
[60]MitwallyMF,CasperRF.Single dose administration of an aromatase Inhibitor for ovarian stimulation.Fertil Steril,2005,83(1):229-31.
[61]BarrosoG, MenocalG, FelixH, eta.l Comparison of the efficacy of the aromatase inhibitor letrozole andclomiphene citrate as adjuvants to recombinant follicle stimulating hormone in controlled ovarian hyperstimu-lation:a prospective, randomized,blinded clinical trial [J]. Fertil Steri,l 2006,86 (5): 1428-1431.
[62]薛白,王秀英.来曲唑用于氯米芬治疗失败的不孕症76例临床疗效观察[J].徐州医学院学报,2008,28(2)：105.
[63]徐凤琴,张云山.多囊卵巢综合征患者临床促排卵转体外受精-胚胎移植分析[J].天津医药,2008,4(36)：261-263.
[64]吴成平,谷玉,李静,赵芳.体外受精-胚胎移植治疗PCOS不孕分析[J].中国优生与遗传杂志.2009,17(1)：91-92.
[65]杨正望,尤昭玲,冯光荣.多囊卵巢综合征与肾虚血瘀关系浅探[J].湖南中医药导报,2004,10(12)：4-5.
[66]杨悦娅.朱南孙治疗多囊卵巢综合征的思路与方法[J].上海中医药杂志,2006,(40)：1.
[67]夏桂成.用动静观指导滋阴补肾调治多囊卵巢综合征[J].江苏中医药,2006,27(3)：12-13.
[68]韩宁.多囊卵巢综合征临证探微[J].江苏中医药,2009,41(4)：31-32.
[69]付灵梅,马红霞.尤昭玲教授论治多囊卵巢综合征的经验[J].中医药信息,2003,20(1)：39-40.
[70]肖承惊.多囊卵巢综合征所致不孕的认识[J].江苏中医药,2006,27(3)：8-9.
[71]董联玲.柴胡疏肝散临床应用体会[J].山西中医,2004,20(1)：40.
[72]刘丽清,蔡平平.戴德英治疗多囊卵巢综合征经验[J].中医杂志,2002,43(4)：261-262.
[73]赵薇,张晓甦,范旭阳,等.肝脾肾失调与多囊卵巢综合征胰岛素抵抗所致不孕的关系[J].陕西中医,2007, 28(7)：855-857.
[74]王晓静.杨鉴冰教授治疗多囊卵巢综合征经验介绍[J].陕西中医学院学报,2009,32(3)：18-19.
[75]尤昭玲.补肾化瘀法治疗多囊卵巢综合征心得[J].江苏中医药,2006,27(3)：11.
[761常文艺.中医辨证治疗多囊卵巢综合症[J].内蒙古中医药,2008.19-20.
[77]赵薇,谈勇.谈勇教授治疗多囊卵巢综合征经验介绍[J].新中医,2010,42(2)：99-100.
[78]王娜,薛秀伟.多囊卵巢综合征不孕的中医治疗[J].中国实用乡村医生杂志,2004,11(4)：35-36.
[79]徐莲薇,倪晓容,叶玉妹,等.补肾活血调周法治疗多囊卵巢综合征征疗效观察[J].上海中医药杂志,2008,42(12)：37-39.
[80]徐佳,曲惠卿,方海琳.电针配合耳穴贴压对肥胖伴多囊卵巢综合征患者血清胰岛素及睾酮的影响[J].中国针灸,2009,29(6)：441-443.
[81]刘桂英.穴位埋线配合健脾补肾化痰中药对肥胖型多囊卵巢综合征患者高雄激素血症的影响[1D].广州：广州中医药大学,2008.
[1]Lee MT, Anderson E, Lee GY. Changes in ovarian morphology and serum hormones in the rat after treatment with dehydroepiandrosterone. Anat Rec,1991,231:185-192.
[2]Anderson E, Lee GY, O.brien K. Polycystic ovarian condition in the dehydroepiandrosterone treated rat model:hyperandrogenism and the resumption of meiosis are major initial events associated with cystogenesis of antral follicles. Anat Rec,1997,249:44-53.
[3]Anderson E, Lee GY. The polycystic ovarian (PCO)conditionrapoptosis and epithelialization of the ovarian antral follicles are aspects of cystogenesis in the dehydroepiandrosterone (DHEA)-treated rat model. Tissue Cell,1997,29:171-189.
[4]徐海波,吴清和.中药血清药理学实验方法的探讨[J].中国中医药科技,2000,7(1)：43-44.
[5]王力倩,李仪奎,符胜光,等.血清药理学方法研究探索[J].中药药理与临床,1997,13(3)：29-31.
[6]李仪奎.中药血清药理学实验方法的若干问题[J].中药新药与临床药理,1999,10(2)：95-98.
[7]韩林.中药血清药理学研究方法探讨与思考[J].浙江中西医结合杂志,2009,19(5)：319-321.
[8]Chadha S,Pache T D,Huikeshoven F J M,et al.Androgen receptors expression in human ovarian and uterine tissue of long term androgen-treated transsexual women.Hum Pathol,1994,25:1198-1204.
[9]Zlotinik A, Yoshie O. Che mokines:A new classification system and their role in immunity [J].Immunity, 2000,12(1):121-127.
[10]Parrott JA, SkinnerMK. Kit ligand/s tern cell factor induces primordial fol licle devel opment and init i at es foil ic ulogenes is [J].Endocrinology,1999,140(9):4262-4271.
[11]Zhao HB, Luo LL, Li u Y. The effec t of insulin like growth factor-I onDNA s ynthesis of c ultured human granulosa cells in vitro [J].Di si Junyi Daxue Xuebao (J Fourth Mil Med Univ),2000,21(3):282-285.
[12]George BJ, Meredith JS,Peter B, et al. Kit signaling via PI3K promotes ovarian follicle maturation but is dispensable for primordial follicle activation.Dev Biol,2009,331(2):292-299.
[13]Parrott JA, Skinner MK.Kit-ligand/stem cell factor induces primordial follicle development and initiates folliculogenesis [J].Endocrinology,1999,140(9):4262-4271.
[14]Tanikawa M, Harada T, Ito M, et al. Presence of stem cell factor in follicular fluid and its expression in the human ovary[J].Fertil Steril,2000,73(6):1259-1260.
[15]Ronen-Fuhrmann T,Timberg R,King SR,et al.Spatio-tern-poral expression patterns of steroidogenic acute regulatory protein during follicular development in the rat ovary. Endocrin,1998,139:303-315.
[16]Bao B,Calder MD,Xie S,et al.Expression of steroidogenic acute regulatory protein messenger ribonucleic acid is limited to theca of healty bovine follicles collected during recruitment selection,and dominance of follicles of the first follicular wave.Bro Repro,1998,59:953-959.
[1]乐杰主编.妇产科学[M].第7版,北京：人民卫生出版社,2008：351.
[2]中华人民共和卫生部.中药新药临床研究指导原则[S].北京：人民卫生出版社,1993：276-279.
[3]夏桂成.用动静观指导滋阴补肾调治多囊卵巢综合征[J].江苏中医药,2006,27(3)：12-13.
[4]尤昭玲,杨正望,傅灵梅.多囊卵巢综合征从肾虚血瘀调治的探讨[J].湖南中医学院学报,2005,25(1)：25-26.
[5]梁瑞宁.女性“任通冲盛”客观指征初探[J].实用中医药杂志.2010.(26)：504-505.
[6]任青玲,谈勇.夏桂成教授阴阳太极八卦图与月经周期演变规律诠释[J].浙江中医药大学报,2006,30(4)：331-334.
[7]张丽梅,侯丽辉.自拟补肾汤配合电针治疗PCOS临床观察.临床杂志,2003,19(4)：23-4.
[8]李炳如.补肾药对下丘脑-垂体-性腺轴功能的影响[J].中医杂志,1984,7：63
[9]张月萍,俞瑾.雄激素致不孕大鼠发病机制及滋肾阴药对其促排卵的作用[J].中华内分泌代谢杂志,1994,10(2)：98-101.
[10]杜惠兰,宋翠淼,马惠荣,等.补肾固冲系列方诱发小鼠排卵的实验研究[J].河北中医药学报,2001,16(3)：8-9.
[11]吕雪梅.补肾对卵泡液代谢组学及ca2+浓度影响的研究[D].山东：山东中医药大学,2007.
[12]连方,孙振高,张建伟等.二至天癸方对小鼠卵细胞质量影啊的实验研究.中国中西医结合杂志,2004,24(7)：625-627.
[13]张艳萍.补肾调冲对PCOS血清、卵泡液TNF-a、IL-6的影响及其与卵细胞质量的相关性研究[D].山东：山东中医药大学,2008.
[14]耿白霞,任伟慧.SCF在PCOS患者卵巢组织中的表达[J].中国误诊学杂志,2009,9(22)：5329-5330.
[15]王桂红,赵海波,辛晓燕.人卵泡液中干细胞因子浓度及其对卵泡发育的影响.第四军医大学报,2003；24(6)：543-545.
[16]赵海波,刘义.人卵泡液中TNF-a和IL-6浓度及其对卵巢功能的影响.第四军医大学学报,2001；22(5)：429-431.
[17]李小平.益肾清肝活血治疗多囊卵巢综合征所致不孕.福建中医学院报,2005,15(4)：9-11.
[18]邵明霞,张学玲,孙娟娟,等.中西医结合治疗多囊卵巢综合征所致不孕的临床观察.四川中医,2006,24(1)：90-1.
[19]黄玉华,赵子厚,柴松岩,等.益肾健脾养血通利方对Poresky PCOS大鼠模型的观察.中国中医基础医学杂志,2004,10(01)：38-41.
[20]Yan W, Suominen J, Toppari J. Stem cell factor protects germ cells from apoptosis in vitro[J]. J Cell Sci, 2000,113:161-168.
[21]Mitsunari M, Harada T, Tanikawa M, et al. The potential role of stem cell factor and its receptor c-kit in the mouse blastocyst implantation[J]. Mol HumReprod,1999,5(9):874-879.
[22]Kauma S, Huff T, Krystal G, et al. The expression of stem cell factor and its receptor, c-kit in human endometrium and placental tissues during pregnancy[J]. Clin Endocrinol Metab,1996,81(3):1261-1266.
[2]Kahsar-MillerMD, Nixon C, Boots LR,et a.1 Prevalence of polycystlc ovary syndrome (PCOS) in first-degree relatives of patients with PCOS.FertilSteri,l 2001,75(1):53-58.
[3]Cooper HE,Spellacy WN,Prem KA,et al.Hereditary factors in the stein-leventhal syndrome.Am J Obstet Gynecol,1968,10:371-387.
[4]曹云霞,庄广伦.多囊卵巢综合征的染色体分析[J].中国优生与遗传杂志,2001,9(4)：37-38.
[5]Norman RJ, Dewailly D, Legro RS, et a.1 Polycystic ovary syndrome.Lancet,2007,370(9588); 685-697.
[6]Diamanti-Kandarakis E, Piperi C. Genetics of polycystic ovary syndrome:searching for the way out of the labyrinth [J]. Hum ReprodUpdate,2005,11(6):631-643.
[7]LegroRS, Strauss JF. Molecular progress in infertility:polycystic ovary syndrome [J]. FertilSteri,l 2002, 78(3):569-576.
[8]JakubowskiL.Genetic aspects ofpolycystic ovary syndrome.Endokry-nolPo,l 2005,56(3):285-293.
[9]李琳PCOS与青春期胰岛素抵抗.实用妇产科杂志,2005,21(9)：522-524.
[10]胡燕宁.少年多囊卵巢综合征与其父母代谢综合征的相关性研究[J].现代中西医结合杂志,2008,17(3)：335-337.
[11]KumarA,WoodsKS, BartolucciA, et a.1 Prevalence of adrenal androgen excess in patientswith the polycystic ovary syndrome(PCOS)[J].Clin Endocrino,l 2005,62(6):644-649.
[12]Gonzalez F,ChangL,HorabT,et al.Evidence for heterogeneous etiologies of adrenal dysfunction in polycystic ovary syndrome[J]. Fertil Steril,1996,66(3):354-361.
[13]Poretsky L, Piper B. Insulin resistance, hypersecretion of LH, and adual defect hypothesis for the pathogenesis of polycystic ovary syndrome[J]. Obstet Gynecol,1994,84(4):613-621.
[14]王蔼明,李美芝.多囊卵巢综合征患者血雄激素与胰岛素、黄体生成素水平关系的探讨[J].生殖医学杂志,2004,13(1)：47-49.
[15]Ludwig AK, Weiss JM, Tauchert S, et al. Influence of hypo-and hyperglycaemia on plasma leptin concentrations in healthy women and in women with polycystic ovary syndrome [J]. Hum Reprod,2007, 22(6):1555-1556.
[16]Fedorcsak P, Raki M, Storeng R. Characterization and depletion of leukocytes from cells isolated from the pre-ovulatory ovarian follicle [J]. Hum Reprod,2007,22(4):989-994.
[17]Stubbs SA, Stark J, Dilworth SM, et al. Abnormal preantral folliculogenesis in polycystic ovaries is associated with increased granulosa cell division [J]. J Clin Endocrinol Metab,2007,92(11):4418-4426.
[18]Webber LJ, Stubbs S, Stark J, et al. Prolonged survival in culture of preantral follicles form polycystic ovaries [J]. J Clin EndocrinolMetab,2007,92(5):1975-1978.
[19]GuthrieHD, GarrettWM, CooperBS. Follicle-stimula-ting hormone and insulin-like growth factor-Ⅰ attenuate apoptosis in cultured porcine granulosa cells [J].Biol Reprod,1998,58(2):390-396
[20]Coffey S,Mason H.The effect of polycystic ovary syndrome on health-related quality of life[J].Gynecol Endocrinol,2003,17(5):379-386.
[21]Elsenbruch S,Hahn S,Kowalsky D,et al.Quality of life,psychosocial well-being,and sexual satisfaction in women with polycystic ovary syndrome[J].J Clin Endocrinol Metab,2003,88(12):5801-5807.
[22]王玉蓉,曹云霞,丛林,等.多囊卵巢综合征患者的调查及心理分析[J].安徽医科大学学报,2005,40(3)：277-279.
[23]Eggerd S,Kirchengast S.The polycystic ovary syndrome a medical condition but also an important psychosocial problem[J].Coll Antropol,2001,25(2):673-685.
[24]Solomon CG.The epidemiology of polycystic ovary syndrome.Prevalence and associated disease risks.EndocrinolMetab Clin North Am,1999,28(2):247.
[25]Dumesic DA,Abbott DH.Implications of polycystic ovary syndrome on oocyte development[J].Semin Reprod Med,2008,26(1):53-61.
[26]Steckler T,Wang J,Bartol FF,et al.Fetal programming:prenatal testosterone treatment causes intrauterine growth retardation,reduces ovarian reserve and increases ovarian follicular recruitment[J]Endocrinology, 2005,146(7):3185-3193.
[27]Gougeon A.Regulation of ovarian follicular development in primates:facts and hypothesis[J].Endocr Rev,1996,17(2):121-155.
[28]kJakimiuk AJ,Weitsman SR,Brzechffa PR,et al.Aromatase mRNA expression in individual follicles from polycystic ovaries[J].MolHum Reprod,1998,4(1):1-8.
[29]D.S.Guzick,Polycystic ovary syndrome[J].Obstet Gynecol,2004,103:181-193.
[30]De Leo VDE, Marca ALA.Petiaglia F. Insulin-lowering agents in the management of polycystic ovary syndrome[J].Endocrine REV,2003,24(5):633-667.
[31]Kurzawa R,Ciepiela P,Baczkowski T, et al.Comparison of embryological and clinical outcome in GnRH antagonist vs.GnRH agonist protocols for in vitro fertilization in PCOS non—besepatients.Aprospective randomized study[J].J Assist Reprod Genet,2008,25(8):365-374.
[32]Vendola KA, Zhou J, Adesanya OO et al.Androgen stimulate early stages of follicular growth in the primate ovary.J Clin Invest,1998,101 (12):2622.
[33]乐杰,谢幸,林仲秋,等.妇产科学[M].第7版.北京：人民卫生出版社,2008：315.
[34]Greisen S,Ledet T,Ovesen P.Lack of association between polycystic ovary syndrome and embryonic aneuploidy[J].Hum Reprod,2001,16(10):2061-2065.
[35]Franks S,Stark J,Hardy K.Follicle dynamics and anovulation in polycystic ovary syndrome[J].Hum Reprod Update,2008,14(4):367-378.
[36]Picton HM,Harris SE,Muruvi W,et al.The in vitro growth and maturation of follicles[J] Reproduction, 2008,136 (6):703-715.
[37]Hillier SG.Current concepts of the roles of follicle stimulating hormone and luteinizing hormone in folliculogenesis[J].Hum Reprod,1994,9(2):188-191.
[38]Balen AH,ConwayGS,HomburgR,LegroRS. Polycystic Ovary Syndrome. A Guide to Clinical Management[J].London,UK:Taylor&Francis,2005,47-67.
[39]Kezele PR,Nilsson EE,Skinner MK.Insulin but not insulin-like growth factor-1 promotes the primordial to primary follicle transition[J].Mol Cell Endocrinol,2002,192(1-2):37-43.
[40]Balen AH,Conway GS,Homburg R,et al.Polycystic Ovary Syndrome-A Guide to Clinical Management [M].London:Taylor and Francis,2005:47-67.
[41]Zhang Y,Proenca R,Maffei M,et al. Positional cloning of the mouse obese gene and its human homologue [J]. Nature,1994,372(6505):425-432.
[42]Erturk E, Kurn N, Savci V et all Serum lep tin levels correlate with pbesity parameters but not with hyperin sulinism in women with polycystic ovary syndromelFertil Steril,2004,82 (5):1364.
[43]Li MG,Ding GL,Chen XJ,et al.Association of serum and follicular fluid leptin concentrations with granulosa cell phosphorylated signal transducer and activator of transcription 3 expression in fertile patients with polycystic ovarian syndrome[J].J Clin Endocrinol Metab,2007,92(12):4771-4776.
[44]Agarwal SK,Vogel K,Weitsman SR,et al.Leptin antagonizes the insulin-like growth factor-1 augmentation of steroidogenesis in granulosa and theca cells of the human ovary[J].J Clin EndocrinolMetab,1999,84 (3):1072-1076.
[45]Magoffin DA. Ovarian enzyme activities in women with Polycystic ovary syndrome. FertilSteril, 2006,86(7):9-11.
[46]NelsonVL,QinKnKN,RosenfieldRL,etal.The biochemical basis for increased testosterone production in Theca cells Propagated from patients with polycystic ovary syndrome[J],ClinEndocrinol Metab,2001, 86(12):5925-33.
[47]Stubbs SA, Hardy K, Da Silva-Buttkus P, et al. Anti-Mullerian hormone protein expression is reduced during the initial stages of follicle development in human polycystic ovaries. J Clin Endocrinol Metab, 2005,90(10):5536-43.
[48]Ehrmann DA,Liljenquist DR,Kasza K,et al.Prevalence and predictors of the metabolic syndrome in women with polycystic ovary syndrome[J].J Clin Endocrinol Metab.2006,91:48.
[49]林琳,刘复权,张晓莉,等Diane-35对多囊卵巢综合征患者肾素-血管紧张素系统及促排卵的影响[J].河北医科大学学报,2005,26(5)：370-372.
[50]Khattab S,Mohsen IA,Foutouh IA,et al.Metformin reduces abortion in pregnant women with polycystic ovary syndrome[J].GynecolEndcrinol,2006,22(12):680.
[51]赵军招,叶碧绿,林金菊,等.二甲双胍在多囊卵巢综合征促排卵治疗中的作用[J].中华妇产科杂志,2003,38(9)：545-548.
[52]The Rotterdam ESHRE/ASRM-Sponsored PCOSWorkshop Group.Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome[J]. Fertil Steri,l 2004,81(1): 19-25.
[53]Thessaloniki ESHRE/ASRM-Sponsored PCOS ConsensusWorkshop Group.Consensus on infertility treatment related to polycystic ovary syndrome[J]. Fertil Steri,l 2008,89(3):505-522.
[54]HolzerH,Casper R,Tulandi T.A new era in ovulation Induction[J].Ferti-lS teri,l 2006,85(2):277-284.
[55]Rossing MAIClomifene is used in women with Polycystic ovary syndrome[J].NEngl J Med,1994,331: 335-339.
[56]乔杰.多囊卵巢综合征的促排卵治疗[J].北京医科大学学报.1997,29(2)：190-192.
[57]张娜,等.氯米芬联合促卵泡生长激素低量缓增方案应用于多囊卵巢综合征效果观察[J].郑州大学学报(医学版).2007,42(4)：682-684.
[58]Tiboni G M.Aromatase inhibitors and teratogenesis[J].Fertility and Sterility,2004,81(4):1158-1159.
[59]Robert F,Mohamed F,Mitwally M,et al.Aromatase inhibitors for ovulation induction[J].J Clin Endocrinol Metabol,2005,91(3):760-761.
[60]MitwallyMF,CasperRF.Single dose administration of an aromatase Inhibitor for ovarian stimulation.Fertil Steril,2005,83(1):229-31.
[61]BarrosoG, MenocalG, FelixH, eta.l Comparison of the efficacy of the aromatase inhibitor letrozole andclomiphene citrate as adjuvants to recombinant follicle stimulating hormone in controlled ovarian hyperstimu-lation:a prospective, randomized,blinded clinical trial [J]. Fertil Steri,l 2006,86 (5): 1428-1431.
[62]薛白,王秀英.来曲唑用于氯米芬治疗失败的不孕症76例临床疗效观察[J].徐州医学院学报,2008,28(2)：105.
[63]徐凤琴,张云山.多囊卵巢综合征患者临床促排卵转体外受精-胚胎移植分析[J].天津医药,2008,4(36)：261-263.
[64]吴成平,谷玉,李静,赵芳.体外受精-胚胎移植治疗PCOS不孕分析[J].中国优生与遗传杂志.2009,17(1)：91-92.
[65]杨正望,尤昭玲,冯光荣.多囊卵巢综合征与肾虚血瘀关系浅探[J].湖南中医药导报,2004,10(12)：4-5.
[66]杨悦娅.朱南孙治疗多囊卵巢综合征的思路与方法[J].上海中医药杂志,2006,(40)：1.
[67]夏桂成.用动静观指导滋阴补肾调治多囊卵巢综合征[J].江苏中医药,2006,27(3)：12-13.
[68]韩宁.多囊卵巢综合征临证探微[J].江苏中医药,2009,41(4)：31-32.
[69]付灵梅,马红霞.尤昭玲教授论治多囊卵巢综合征的经验[J].中医药信息,2003,20(1)：39-40.
[70]肖承惊.多囊卵巢综合征所致不孕的认识[J].江苏中医药,2006,27(3)：8-9.
[71]董联玲.柴胡疏肝散临床应用体会[J].山西中医,2004,20(1)：40.
[72]刘丽清,蔡平平.戴德英治疗多囊卵巢综合征经验[J].中医杂志,2002,43(4)：261-262.
[73]赵薇,张晓甦,范旭阳,等.肝脾肾失调与多囊卵巢综合征胰岛素抵抗所致不孕的关系[J].陕西中医,2007, 28(7)：855-857.
[74]王晓静.杨鉴冰教授治疗多囊卵巢综合征经验介绍[J].陕西中医学院学报,2009,32(3)：18-19.
[75]尤昭玲.补肾化瘀法治疗多囊卵巢综合征心得[J].江苏中医药,2006,27(3)：11.
[761常文艺.中医辨证治疗多囊卵巢综合症[J].内蒙古中医药,2008.19-20.
[77]赵薇,谈勇.谈勇教授治疗多囊卵巢综合征经验介绍[J].新中医,2010,42(2)：99-100.
[78]王娜,薛秀伟.多囊卵巢综合征不孕的中医治疗[J].中国实用乡村医生杂志,2004,11(4)：35-36.
[79]徐莲薇,倪晓容,叶玉妹,等.补肾活血调周法治疗多囊卵巢综合征征疗效观察[J].上海中医药杂志,2008,42(12)：37-39.
[80]徐佳,曲惠卿,方海琳.电针配合耳穴贴压对肥胖伴多囊卵巢综合征患者血清胰岛素及睾酮的影响[J].中国针灸,2009,29(6)：441-443.
[81]刘桂英.穴位埋线配合健脾补肾化痰中药对肥胖型多囊卵巢综合征患者高雄激素血症的影响[1D].广州：广州中医药大学,2008.
[1]Lee MT, Anderson E, Lee GY. Changes in ovarian morphology and serum hormones in the rat after treatment with dehydroepiandrosterone. Anat Rec,1991,231:185-192.
[2]Anderson E, Lee GY, O.brien K. Polycystic ovarian condition in the dehydroepiandrosterone treated rat model:hyperandrogenism and the resumption of meiosis are major initial events associated with cystogenesis of antral follicles. Anat Rec,1997,249:44-53.
[3]Anderson E, Lee GY. The polycystic ovarian (PCO)conditionrapoptosis and epithelialization of the ovarian antral follicles are aspects of cystogenesis in the dehydroepiandrosterone (DHEA)-treated rat model. Tissue Cell,1997,29:171-189.
[4]徐海波,吴清和.中药血清药理学实验方法的探讨[J].中国中医药科技,2000,7(1)：43-44.
[5]王力倩,李仪奎,符胜光,等.血清药理学方法研究探索[J].中药药理与临床,1997,13(3)：29-31.
[6]李仪奎.中药血清药理学实验方法的若干问题[J].中药新药与临床药理,1999,10(2)：95-98.
[7]韩林.中药血清药理学研究方法探讨与思考[J].浙江中西医结合杂志,2009,19(5)：319-321.
[8]Chadha S,Pache T D,Huikeshoven F J M,et al.Androgen receptors expression in human ovarian and uterine tissue of long term androgen-treated transsexual women.Hum Pathol,1994,25:1198-1204.
[9]Zlotinik A, Yoshie O. Che mokines:A new classification system and their role in immunity [J].Immunity, 2000,12(1):121-127.
[10]Parrott JA, SkinnerMK. Kit ligand/s tern cell factor induces primordial fol licle devel opment and init i at es foil ic ulogenes is [J].Endocrinology,1999,140(9):4262-4271.
[11]Zhao HB, Luo LL, Li u Y. The effec t of insulin like growth factor-I onDNA s ynthesis of c ultured human granulosa cells in vitro [J].Di si Junyi Daxue Xuebao (J Fourth Mil Med Univ),2000,21(3):282-285.
[12]George BJ, Meredith JS,Peter B, et al. Kit signaling via PI3K promotes ovarian follicle maturation but is dispensable for primordial follicle activation.Dev Biol,2009,331(2):292-299.
[13]Parrott JA, Skinner MK.Kit-ligand/stem cell factor induces primordial follicle development and initiates folliculogenesis [J].Endocrinology,1999,140(9):4262-4271.
[14]Tanikawa M, Harada T, Ito M, et al. Presence of stem cell factor in follicular fluid and its expression in the human ovary[J].Fertil Steril,2000,73(6):1259-1260.
[15]Ronen-Fuhrmann T,Timberg R,King SR,et al.Spatio-tern-poral expression patterns of steroidogenic acute regulatory protein during follicular development in the rat ovary. Endocrin,1998,139:303-315.
[16]Bao B,Calder MD,Xie S,et al.Expression of steroidogenic acute regulatory protein messenger ribonucleic acid is limited to theca of healty bovine follicles collected during recruitment selection,and dominance of follicles of the first follicular wave.Bro Repro,1998,59:953-959.
[1]乐杰主编.妇产科学[M].第7版,北京：人民卫生出版社,2008：351.
[2]中华人民共和卫生部.中药新药临床研究指导原则[S].北京：人民卫生出版社,1993：276-279.
[3]夏桂成.用动静观指导滋阴补肾调治多囊卵巢综合征[J].江苏中医药,2006,27(3)：12-13.
[4]尤昭玲,杨正望,傅灵梅.多囊卵巢综合征从肾虚血瘀调治的探讨[J].湖南中医学院学报,2005,25(1)：25-26.
[5]梁瑞宁.女性“任通冲盛”客观指征初探[J].实用中医药杂志.2010.(26)：504-505.
[6]任青玲,谈勇.夏桂成教授阴阳太极八卦图与月经周期演变规律诠释[J].浙江中医药大学报,2006,30(4)：331-334.
[7]张丽梅,侯丽辉.自拟补肾汤配合电针治疗PCOS临床观察.临床杂志,2003,19(4)：23-4.
[8]李炳如.补肾药对下丘脑-垂体-性腺轴功能的影响[J].中医杂志,1984,7：63
[9]张月萍,俞瑾.雄激素致不孕大鼠发病机制及滋肾阴药对其促排卵的作用[J].中华内分泌代谢杂志,1994,10(2)：98-101.
[10]杜惠兰,宋翠淼,马惠荣,等.补肾固冲系列方诱发小鼠排卵的实验研究[J].河北中医药学报,2001,16(3)：8-9.
[11]吕雪梅.补肾对卵泡液代谢组学及ca2+浓度影响的研究[D].山东：山东中医药大学,2007.
[12]连方,孙振高,张建伟等.二至天癸方对小鼠卵细胞质量影啊的实验研究.中国中西医结合杂志,2004,24(7)：625-627.
[13]张艳萍.补肾调冲对PCOS血清、卵泡液TNF-a、IL-6的影响及其与卵细胞质量的相关性研究[D].山东：山东中医药大学,2008.
[14]耿白霞,任伟慧.SCF在PCOS患者卵巢组织中的表达[J].中国误诊学杂志,2009,9(22)：5329-5330.
[15]王桂红,赵海波,辛晓燕.人卵泡液中干细胞因子浓度及其对卵泡发育的影响.第四军医大学报,2003；24(6)：543-545.
[16]赵海波,刘义.人卵泡液中TNF-a和IL-6浓度及其对卵巢功能的影响.第四军医大学学报,2001；22(5)：429-431.
[17]李小平.益肾清肝活血治疗多囊卵巢综合征所致不孕.福建中医学院报,2005,15(4)：9-11.
[18]邵明霞,张学玲,孙娟娟,等.中西医结合治疗多囊卵巢综合征所致不孕的临床观察.四川中医,2006,24(1)：90-1.
[19]黄玉华,赵子厚,柴松岩,等.益肾健脾养血通利方对Poresky PCOS大鼠模型的观察.中国中医基础医学杂志,2004,10(01)：38-41.
[20]Yan W, Suominen J, Toppari J. Stem cell factor protects germ cells from apoptosis in vitro[J]. J Cell Sci, 2000,113:161-168.
[21]Mitsunari M, Harada T, Tanikawa M, et al. The potential role of stem cell factor and its receptor c-kit in the mouse blastocyst implantation[J]. Mol HumReprod,1999,5(9):874-879.
[22]Kauma S, Huff T, Krystal G, et al. The expression of stem cell factor and its receptor, c-kit in human endometrium and placental tissues during pregnancy[J]. Clin Endocrinol Metab,1996,81(3):1261-1266.