摘要
目的:探讨体外受精-胚胎移植(IVF-ET)降调节后的证候特点,及证候与子宫内膜容受性的相关性。
方法:临床研究对200例行IVF-ET的患者,采用流行病学调查方法,结合聚类分析总结IVF-ET降调节后的中医证候特点;同时应用以药测证的方法,对60例行IVF-ET的患者进行系统的临床观察,其中中药治疗组30例,对照组30例。分别对两组患者中医证候积分变化、子宫内膜血流、优质卵率、受精率、优质胚胎率、妊娠率及临床妊娠率进行评定。实验研究分别采用GnRH-a降调节小鼠模型和GnRH-a超排卵小鼠模型,与正常小鼠及加用二至天癸颗粒小鼠对照,观察对阴道涂片、子宫、卵巢指数、子宫、卵巢组织形态学、血浆cAMP、cGMP含量、子宫内膜HOXA10mRNA表达的影响。
结果:聚类分析结果表明IVF-ET降调节后主要表现为肾虚证,以肾阴虚为主,兼有肾阳虚症状。经补肾中药二至天癸颗粒治疗后,可明显改善患者肾虚症状。GnRH-a降调节后,小鼠体重下降;阴道涂片表现动情间期;子宫、卵巢指数下降;子宫内膜厚度明显变薄,腺体显著减少,部分腺体萎缩消失;卵泡层次紊乱,部分卵泡闭锁,部分卵泡细胞坏死,炎细胞浸润,部分间质纤维组织增生;cAMP含量及cAMP/cGMP比值明显升高;HOXA10基因表达下调;二至天癸颗粒可改善上述指标;从微观角度证实其肾虚本质。二至天癸颗粒还可通过改善子宫内膜血流及GnRH-a超排卵小鼠着床期子宫内膜组织形态学指标、提高cAMP含量、上调HOXA10基因发挥改善子宫内膜容受性的作用。
结论:体外受精-胚胎移植降调节后主要表现为肾虚证,以肾阴虚为主,补肾中药二至天癸颗粒可提高子宫内膜容受性,表明了肾虚与子宫内膜容受性的相关性。为中医药在IVF-ET的应用提供依据,丰富了中医肾主生殖的理论内涵。
Objective: To study the characteristics of Traditional Chinese Medical syndrome after in vitro fertilization and embryo transfer (IVF-ET) down regulation and the relationship between Traditional Chinese Medical syndrome and endometrial receptivity.
Methods: In clinical study, 200 infertile patients undergoing IVF-ET were studied using epidemiologic method and cluster analysis, and the method of syndrome differentiation by effects of drug was also employed to determine the characteristics of Traditional Chinese Medical syndrome after IVF-ET down regulation. In experimental study, the mice model of GnRH-a down regulation and GnRH-a assisted controlled ovarian hyperstimulation were adopted, compared with the normal group and the TCM group. Vaginal smear, the index of uterine and ovary were observed; the endometrial and ovarian morphology were investigated by light microscope; the concentration of cAMP and cGMP were measured by radioimmunoassay (RIA) and the expression of HOXA10 in the endometrium was detected by in situ hybridization method.
Results: According to the cluster analysis, the characteristics of Traditional Chinese Medical Syndrome of IVF-ET down regulation were identified as deficiency of kidney which were mainly yin deficiency of kidney combined with the symptom of yang deficiency of kidney. And symptoms of Shenxu had been significantly improved by taking Erzhi Tiangui Granules. In experimental study, the characteristics were also proved. And Erzhi Tiangui Granules can improve the endometrial receptivity by enhancing the blood perfusion in the endometrium, ameliorating morphological index in mice endometrial tissue and increasing the concentration of cAMP and expression of HOXA10.
Conclusion: The characteristics of Traditional Chinese Medical syndrome of IVF-ET down regulation are identified as deficiency of kidney which are mainly yin deficiency of kidney, and the endometrial receptivity can be improved by tonifying kidney, which indicated the relationship between the deficiency of kidney and the endometrial receptivity.
引文
[1] 史小林.人类生殖学,北京:科学出版社,2002:299.
[2] 黄荷凤.现代辅助生育技术,北京:人民军医出版社,2003:341.
[3] Pantos K, Meimeth-Damianaki T, Vaxevanoglou T, et al. Prospective study of a modified gonadotropin-releasing hormone agonist long protocol in an in vitro fertilization program. Fertil Steril, 1994;61(4):709-713.
[4] Lopata A. Implantation of the human embryo. Hum Reprod, 1996;11:175-184.
[5] Mirkin S, Nikas G, Hsim JG , et al. Gene.expression profiles and structural/functional features of the pri-implantation endometrium in natural and gonadotropin-simulated cycles. J Clin Endocrinol Metab,2004;89(11):5742-5752.
[6] 金方.何嘉琳谈在辅助生育技术中中医药的应用.北京中医,2004;23(6):329-330.
[7] 谈勇,石川睦男.补肾调周法在体外受精-胚胎移植前应用的临床观察.中国中医药信息杂志,2001;8(12):56-57.
[8] 连方,张建伟,张宁,等.中西医结合疗法在试管婴儿技术中的应用.山东中医药大学学报,2002;26(3):182-183.
[9] 张明敏,黄光英,陆付耳,等.补肾益气活血汤对多次助孕技术失败患者结局的影响.微循环学杂志,2002;12 (2):10-12.
[10] 马景兰,马建新,王旭初,等.促孕丸在“试管婴儿”中的应用.中国中西医结合杂志,2005;25 (6):547-548.
[11] 朱文杰,李雪梅,陈秀敏,等.滋肾育胎丸对体外受精-胚胎移植患者胚胎种植率的影响.中国中西医结合杂志,2002;22(10):729-730.
[12] 连方,张宁,张建伟,等.中药配合体外受精及胚胎移植治疗不孕症2例.中国中西医结合杂志,2002;26(3)∶627-628.
[13] 连方,孙振高,张建伟,等.二至天癸方对小鼠卵细胞质量影响的实验研究.中国中西医结合杂志,2004;24(7):625-627.
[14] 连方,孙振高,穆琳,等.二至天癸颗粒提高卵细胞质量与小鼠卵巢内IGF-1R mRNA表达量关系的研究.中国中西医结合杂志,2006;26(5):431-434.
[15] 连方,滕依丽,张建伟,等.二至天癸颗粒联合体外受精-胚胎移植治疗不孕症61例临床研究.中医杂志,2006;47(6):439-441.
[16] 连方,王琳,张建伟,等.二至天癸方对高龄不孕妇女卵巢反应性的影响.中国中西医结合杂志,2006;26(8):685-688.
[17] 刘平,童瑶.中医药科学研究思路与方法,上海:上海中医药大学出版社,2003:156-158.
[18] 乐杰.妇产科学,北京:人民卫生出版社,2004,第六版:381.
[19] 张玉珍.中医妇科学,北京:中国中医药出版社,2005:320.
[20] 郑筱萸.中药新药临床研究指导原则,北京:中国医药科技出版社,2002:385-390.
[21] 沈自尹,王文健.中医虚证辨证参考标准.中国中西医结合杂志,1986;6(10):598.
[22] 胡立胜,周强.中医临床研究设计与SAS编程统计分析.北京:学苑出版社,2004:68.
[23] 梁茂新.中医症状量化的方法初探.中国医药学报,1999;(3):37.
[24] 金丕焕.医用统计方法,上海:上海医科大学出版社,1993:290-298.
[25] 潘永苗,石一复.GnRH-a超排卵对小鼠子宫内膜形态学和雌、孕激素受体表达的影响.生殖与避孕,2002;22(4):195-198.
[26] 章汉旺,高娟.控制性超排卵小鼠围着床期子宫内膜容受性的研究.中国妇幼保健,2005;20(11):1324-1326.
[27] 高娟,章汉旺.GnRH-a 超排卵对小鼠着床期子宫内膜厚度及整合素αv、β1表达的影响.现代妇产科进展,2005;14(2):135-137.
[28] 邹移海,徐志伟,苏钢强.实验动物学,北京:科学出版社,2002,第二版:72,194.
[29] 何清波,苏炳华,钱元.医学统计学及其软件包[M],上海:上海科学技术文献出版社,2003:285.
[30] 胡镜清,赖世隆.中医临床研究方法指南—概论,国家中医药管理局科技教育司,1999:1.
[31] 何裕民.体质聚类研究与辨证体系重建[J].中医研究,1996;9(5):1.
[32] 李秀昌,张红,周喆，等．月经过多证候学的系统聚类分析[J].长春中医学院学报,1999;15(3):33.
[33] 李国春,史欣德.半夏泻心汤临床案例用药量的聚类分析.中医药学刊,2005;23(5):836-838.
[34] 周铭心.用计量化方法探讨《傅青主女科》方药特点及作者真伪[J].中国中医基础医学杂志,1996;2(6):55.
[35] 陈凯佳,肖莹.近现代名老中医治疗“闭经”用药规律初探.中医药学刊,2005;23(10):1831.
[36] 高为民,柳长峰,吕金风,等.模糊聚类法在人参各部位成分鉴别分类中的应用[J].哈尔滨医药,1995;15(1):62.
[37] 汪学昭,毖鹤鸣.女贞子微量元素的模糊聚类分析[J].第二军医大学学报,1995;16(2):183.
[38] 李水福,胡清宇.紫外光谱聚类分析在中药研究中的应用[J].时珍国药研究,1997;8(1):31.
[39] 同[17]:149.
[40] 宓鹤鸣,郭澄,宋宏涛,等.三种菟丝子补肾壮阳作用的比较.中草药,1991;22(12):547-549.
[41] 朱金凤,余运初,周楚华.寿胎丸加味治疗先兆流产的临床观察及实验研究.中西医结合杂志,1987;7(7):407-409.
[42] 李炳如,余运初.补肾药对下丘脑-垂体-性腺轴功能的影响.中医杂志,1984;25(7):63-65.
[43] 王玲,杜守英,钱玉昆.枸杞多糖的免疫调节研究进展.上海免疫学杂志,1995;15(2):118-120.
[44] 周金黄,王筠默.中药药理学,上海:上海科学技术出版社,1986:272.
[45] 同[44]:268.
[46] 同[44]:265.
[47] 同[44]:190.
[48] 同[44]:179.
[49] 陈小野.实用中医证候动物模型学,北京:北京医科大学中国协和医科大学联合出版社,1993:100-144.
[50] 张鹏,洪宝源.人体内 cAMP 与 cGMP 动态平衡机理的探讨.生物化学与生物物理进展,1985;(3):46-48.
[51] 邝安堃等．阴虚病人内分泌、免疫和环核苷酸变化的初步观察．中华内科杂志，1979;18(2):105.
[52] 邝安堃等．阳虚（甲减）和阴虚（甲亢）病人血浆核苷酸的对比．中医杂志，1979;20(7):21.
[53] 邝安堃等．中医分型病人的血浆环核苷酸变化．上篤幸揭┰又荆?980;(3):2.
[54] 夏宗勤,朱文,胡雅儿,等.中医“虚证”理论的初步探讨.中医杂志,1979;20(11):2-9.
[55] 廖菡,张秋俊,马骏,等.葆春方对老年大鼠器官及脑组织中环核苷酸含量的影响.标记物分析与临床,1997;4(4):213-216.
[56] 白润江,马端端,王嘉军,等.香菇多糖对小鼠血浆、胸腺、脾脏 cAMP、cGMP含量的影响.西安医科大学学报,1997;18(1):58-59.
[57] 李震,仇祯绪,李军山,等.自拟补气养血方抗肿瘤机理的研究.实用中西医结合杂志,1997;10(3):257-259.
[58] Benson GV, Lim H, Paria BC, et al. Mechanisms of reduced fertility in Hoxa-10 mutant mice: uterine homeosis and loss of maternal Hoxa-10 expression [J]. Development, 1996; 122(9): 2687-2696.
[59] Bagot CN, Kliman HJ, Taylor HS. Maternal Hoxa10 is required for pinopod formation in the development of mouse uterine receptivity to embryo implantation [J]. Dev Dyn, 2001; 222 (3):538-544.
[60] Bagot CN, Troy PJ, Taylor HS. Alteration of maternal Hoxa10 expression by in vivo gene transfection affects implantation [J]. Gene Ther, 2000; 7(16):1378-1384.
[61] Daftary GS, Taylor HS. Pleiotropic effects of Hoxa10 on the functional development of peri-implantation endometrium[J]. Mol Reprod Dev, 2004; 67(1):8-14.
[62] 高天祥,田竟生.医学分子生物学,北京:科学出版社,2002:145.
[63] 常宇.中医证候基因组学研究的思路—访第一军医大学罗仁教授[J].中国中医药报,2002;1:9.
[64] 罗仁,成玉斌,薛耀明,等.小四五汤治疗 DN 疗效与 ACE 基因相关性研究[J].上海中医药大学学报,2001;15(1):24-26.
[65] 孙伟正,赵淑英,孙岸,等.慢性再生障碍性贫血中医辨证分型与 HLA 基因多肽性相关性的研究[J].中医药信息杂志,2000;3:60-61.
[66] 吴志奎.益髓生血灵治疗β地中海贫血症 29 例临床观察.中医杂志,2000;41(2):90-91.
[67] 王建华,王晓贞.中医证候学研究的思路和方法.北京中医药大学学报,1999;22(4):13-15.
[68] 王红玉.中医证候临床研究进展.中国医学学报,2003;18(12):741-743.
[69] 同[17]:148.
[70] 陈小野.证候动物模型诊断依据的设想与评价.中国医药学报,1987;2(1):50.
[71] 张家庆,杜茹竹.中西医结合研究思路与方法学.上海:上海科技出版社,1985:430.
[72] 贡雪灏,乐桂蓉,张青萍,等.经阴道彩色多普勒超声在体外授精-胚胎移植中的应用.中华超声影像学杂志,2000;9(8):499-500.
[73] Goswamy RK, Steptoe PC. Doppler ultrasound studies of the uterine artery in spontaneous ovarian cycles. Human Reprod, 1988; 3:721-726.
[74] Steer CV, Mason BA, Tan SL, et al. Vaginal color Doppler assessment of uterine artery impedance correlates with immunohistochemical markers of endometrial receptivity required for the implantation of an embryo. Fetil Steril, 1995; 63:101-108.
[75] Kupesic S, urjak A. Uterine and ovarian perfusion during theperiovulatory period assessed by transvaginal color Doppler. Fertil Steril, 1993; 60:439-443.
[76] Kupesic S, Bekavac I, Bjelos D, et al. Assessment of endometrial receptivity by transvaginal color Doppler and three dimensional power Doppler ultrasonography in patients undergoing in vitro fertileization procedures. J Ultrasound Med, 2001; 20(2):125-134.
[77] 谭季春,李守柔,曲文玉,等.彩超观察子宫动脉及螺旋动脉能量图与体外受精的关系.中国超声诊断杂志,2002;3(12):945-947.
[78] Chui DK, Pugh ND, Walker SM, et al. Follicular vascularity-the predictive value of transvaginal power Doppler ultrasonography in an IVF programme: a preliminary study. Hum Reprod, 1997;12:191-196.
[79] 张奕民.肾虚型不孕症和宫内灌注不良的相关性研究.江苏中医,1999;20(1):14-15.
[80] 陈秋梅,张树成,沈明秀.调经孕育方药对排卵障碍性不孕者卵巢、子宫组织血流动力学参数的影响.中国中医药信息杂志,2003;10(12):6-9.
[81] 陈秋梅,张树成,沈明秀.调经孕育方药对排卵障碍性不孕者同步测试的卵泡和子宫内膜生长发育、血流特性的影响.中国实验方剂学杂志,2004;10(2):59-61.
[82] 张扬.控制性超促排卵影响子宫内膜接受性的研究进展.国外医学·妇产科学分册,2003;30(5):295-297.
[83] 栾少红,孙宝治,戴淑真.促排卵药物对大白鼠子宫内膜形态影响的研究.中原医刊,2003;30(12):1-3.
[84] 潘永苗.促排卵药物对子宫内膜形态学的影响.国外医学·妇产科学分册,1999;26(6):340-343.
[85] 罗颂平.中医临床家·罗元恺.北京:中国中医药出版社,2001:86.
[86] 张树成,张志洲,刘效群等.补肾调经方药促进人着床期子宫内膜同步化的组织形态学观察.中国中医基础医学杂志,2002;8(4):48-49.
[87] 张树成,张志洲,刘彬,等.补肾调经方调经促排卵健内膜作用的临床实验研究.中医药学刊,2002;20(6):720-721.
[88] 张树成,沈明秀,吴志奎.补肾生血和补肾调经方药对老龄雌性金黄地鼠生殖器官组织形态的影响.中国民间疗法,1998;6(5):56-57.
[89] 陈秋梅,张树成,贺彬等.调经孕育方药对排卵障碍性不孕者子宫内膜感受态的影响—组织形态学分析.中医药学刊,2004;22(3):464-467.
[90] 王素霞,孙玉英.安胎合剂对 GnRHa 超排卵小鼠子宫内膜形态学的影响.中医药学报,2006;34(5):45-47.
[91] 同[2]:330.
[92] Benson GV, Nguyen THE, Maas RL. The expression pattern of the murine Hoxa-10 gene and the sequence recognition of its homeodomain reveal specific properties of Abdominal B-like genes. Mol Cell Biol.1995; 15(3): 1591-1601.
[93] 李红,陈士岭.HOXA10 基因在子宫内膜组织中的表达及与不明原因不孕的关系[J].中华妇产科杂志,2002;37∶30-32.
[94] Cermik D, Karaca M, Taylor HS. HOXA10 expression is repressed by progesterone in the myometrium: differential tissue-specific regulation of HOX gene expression in the reproductive tract [J]. J Clin Endocrinol Metab,2001;86∶3387-3392.
[95] Yao MW,Lim H,Schust DJ,el al.Gene expression profiling reveals progesterone-mediated cell cycle and immunoregulatory roles of Hoxa-10 in the preimplantation uterus.Mol Endocrinol,2002;17(4):6l0-627.
[96] Taylor HS,Bagot C,Kardana A,et al. HOX gene expression is altered in the endometrium of women with endometriosis.Hum Reprod,1999;14(5):1328-1331.
[97] Cermik D, Selam B, Taylor HS. Regulation of HOXA10 expression by testosterone in vitro and in the endometrium of patients with polycystic ovary syndrome[J]. J Clin Endocrinol Metab, 2003;88∶238-243.
[98] Lessey BA,Castelbaum AJ,Sawin SW,et a1.Aberrant integrin expression in the endometrium of women with endometriosis.Clin Endocrinol Metab,1994;79(2):643-649.
[99] Lim H,Ma L,Ma WQ,etal.HOXA10 regulates uterine stromal cell responsi-veness to progesterone during implantation and decidualization in the mouse[J].Mol Endocrinol,1999;13∶1005-1017.
[100] Qian K,Chen H & Zhu GJ.Differentiation of endometrial stromal cells in vitro:down-regulation of suppression of the cell cycle inhibitor p57 by HOXA10. Mol Hum Reprod, 2005;11(4):245-251.
[101] Gui Y, Zhang J, Yuan L. Regulation of HOXA10 and its expression in normal and abnormal endometrium [J].Mol Hum Reprod,1999;5∶866-873.
[102] Daftary GS, Taylor HS. Pleiotropic effects of HOXA10 on the functional development of peri-implantation endometrium[J]. Mol Reprod Dev, 2004; 67∶8.
[103] Lessey BA, Gui Y, Apparao KB, et al. Regulated expression of heparin-binding EGF-like growth factor (HB-EGF) in the human endometrium: a potential paracrine role during implantation [J]. Mol Reprod Dev, 2002; 62∶446-455.
[104] 张翠珍,王天芳.cAMP、cGMP 拮抗性代谢调节与中医证候关系研究进展.北京中医药大学学报,1999;22(6):51-53.
[1] Lessey BA. Two pathways of progesterone action in the human endometrium: implications for implantation and contraception. Steroids, 2003; 68 (10-13): 809-815.
[2] Tesarik J, Hazout A, Mendoza C. Luteinizing hormone affects uterine receptivity independently of ovarian function. Reprod Biomed Online, 2003; 7(1):59-64.
[3] Stewart CL, Kaspar P, Brunt LJ, et al. Blastocyst implantation depends on maternal expression of leukemia inhibitory factor. Nature, 1992; 359:76-79.
[4] Cheng JG, Rodriguez CI, Stewart CL. Control of Uterine Receptivity and Embryo Implantation by Steroid Hormone Regulation of LIF Production and LIF Receptor Activity: Towards a Molecular Understanding of The Window of Implantation. Rev Endocr Metab Disord, 2002; 3:119-126.
[5] Boucher A, Kharfi A, Al-Akoum M, et al.Cycle-dependent expression of interleukin-1 receptor type II in the human endometrium.Biol Reprod, 2001; 65:890-898.
[6] Song H, Lim H, Das SK, et al. Dysregulation of EGF family of growth factorsand COX-2 in the uterus during the preattachment and attachment reactions of the blastocyst with the luminal epithelium correlates with implantation failure in LIF-deficient mice. Mol EndocrinolV 2000; 14 (8) :1147-1161.
[7] Han VK, Carter AM. Spatial and temporal patterns of expression of messenger RNA for insulin -like growth factors and their binding proteins in the placenta of man and laboratory animals. Placenta, 2000; 21 (4) :289 -305.
[8] Dominguez F, Avila S, Cervero A, et al. A combined approach for gene discovery identifies insulin-like growth factor-binding protein related protein-1 as a new gene implicated in human endometrial receptivity. J Clin Endocrinol Metab, 2003; 88(4): 1849-1857.
[9] Smith SK. Angiogenesis and implantation. Hum Reprod, 2000; 15 [suppl]: 16:59-66.
[10] Kariya M, Kanzaki H, Hanamura T,et al. Progesterone-dependent secretion of macrophage colony-stimulating factor by human endometrial stromal cells of nonpregnant uterus in culture. J Clin Endocrind Metab, 1994; 79(1):86-90.
[11] Tabibzadeh S, Mason JM, Shea W, et al. Dysregulated expression of ebaf, a novel molecular defect in the endometria of patients with infertility. J Clin Endocrinol Metab, 2000; 85(7):2526-2536.
[12] Thomas K. Endometrial integrin expression in women undergoing in vitro fertilization and the association with subsequent treatment outcome. Fertil Steril, 2003; 80 (3):502-507.
[13] Lessey BA. Implantation defects in inferile women with endometriosis.Ann NY Acad Sci, 2002; 955 (3): 265-280.
[14] PalominoWA, Fuentes A, Gonzalez RR, et al. Differential expression of endometrial integrins and progesterone receptor during the window of implantation in normo-ovulatory women treated with clomiphene citrate. Fertil Steril, 2005; 83 (3) : 587-593.
[15] Kumar S, Zhu LJ, Polihronis MK, et al. Progesterone induces calcitonin gene expression in human endometrium within the putative window ofimplantation. J Clin Endocrinol Metab, 1998; 83(12):4443-4450.
[16] Genbacev OD, Prakobphol A, Foulk RA, et al. Trophoblast L-selectin -mediated adhesion at the maternal-fetal interface. Science, 2003;299 (5605)405-408.
[17] Lindhard A,Bentin-Ley U,Ravn V, et al. Biochemical evaluation of endometrial function at the time of implantation. Fertil Steril, 2002; 78 (2):221-233.
[18] Damario MA,Lesnick TG,Lessey BA,et al. Endometrial markers of uterine receptivity utilizing the donor oocyte model. Hum Reprod, 2001; 16 (9):1893-1899.
[19] Spencer TE, Bazer FW. Biology of progesterone action during pregnancy recognition and maintenance of pregnancy. Front Biosci, 2002; 7:d1879-1898.
[20] Bagot CN, Kliman HJ, Taylor HS. Maternal Hoxa10 is required for pinopod formation in the development of mouse uterine receptivity to embryo implantation. Dev Dyn, 2001; 222(3):538-544.
[21] Daftary GS, Troy PJ, Bagot CN, ET al. Direct regulation of beta3-integrin subunit gene expression by HOXA10 in endometrial cells. Mol Endocrinol, 2002; 16(3):571-579.
[22] Troy PJ, Daftary GS, Bagot CN, et a1.Transcriptional repression of peri-Implantation EMX2 expression in mammalian reproduction by HOXA 10.Mol Cell Biol, 2003; 23(1):1-13.
[23] Taylor HS, Fei XL.Emx2 regulates mammalian reproduction by altering endometrial cell proliferation. Mol Endocrinol, 2005; 19(11):2839-2846.
[24] Nikas G, Aghajanova L. Endometrial pinopodes: some more understanding on human implantation[J]. Reprod Biomed Online, 2002; 4(3):18-23.
[25] U sadi R S, Murray M J, Bagnell R C, et al. Temporal and morphologic characteristics of pinopod expression across the secretory phase of the endometrial cycle in normally cycling women with proven fertility[J]. Fertil Steril, 2003; 79(4):970-974.
[26] Garcia-Velasco JA, Nikas G, Remohi J, et al. Endometrial receptivity interms of pinopode expression is not impaired in women with endometriosis in artificially prepared cycles. Fertil Steril, 2001; 75(6):1231-1233.
[27] Stavreus E, Nikas G, Sahlin L, et al. Formation of pinopodes in human endometrium is associated with the concentrations of progesterone and progesterone receptors[J].Fertil Steril, 2001; 76(4):782-791.
[28] Nikas G.. Pinopodes as markers of endometrial receptivity in clinical practice. Hum Reprod, 1999; 14[suppl]:2:99-106.
[29] Bentin Ley U, Sjogren A, Nilsson L, et al. Presence of Uterine pinopodes at the embryo-endometrial interface during human implantation in vitro. Hum Reprod, 1999; 14(2):515-520.
[30] Bentin Ley U. Relevance of endometrial pinopodes for human blastocyst implantation[J]. HumReprod, 2000; 15[Suppl]:6:67-73.
[31] Schild RL, Knobloch C, Dorn C, et al. Endometrial receptivity in an in vitro fertilization program as assessed by spiral artery blood flow, endometrial thickness, endometrial volume, and uterine artery blood flow. Fertil Steril, 2001; 75(2):361-366.
[32] Higuchi T, Kanzaki H, Nakayama H, et al. Induction of tissue inhibitor of metalloproteinase 3 gene expression during in vitro decidualization of human endometrial stromal cells.Endocrinology, 1995; 36:4973-4981.
[33] Salamonsen LA. Role of proteases in implantation. Rev Reprod, 1999; 4:11-22.
[34] Marions L, Danielsson KG. Expression of cyclo-oxygenase in human Endometrium during the implantation period. Mol Hum Reprod, 1999; 5:961-965.
[35] Chwalisz K,Garfield RE.Role of nitric oxide in implantation and menstruation. Hum Reprod, 2000; 15 [Suppl]3:96-111.
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[2] 刘艳娟, 黄光英, 杨明炜, 等.健胎液对胚泡着床障碍小鼠雌、孕激素及其受体的影响.中国中药杂志,2005;30(5):373-376.
[3] 吴瑞瑾,周馥贞.养精种玉汤对原因不明不孕患者子宫内膜基质金属蛋白酶 9表达及性激素调节的影响.中国中西医结合杂志,2004;24(4):294-298.
[4] 贺丽娟.健胎液对小鼠围着床期子宫内膜整合素β3表达的影响.云南中医学院学报,2004;27(1):13-15.
[5] 陈秋梅,张树成,贺斌等.调经孕育方药对排卵障碍性不孕者子宫内膜感受态的影响.中国中医药信息杂志,2004;11(7):576-579.
[6] 蒋晓芳,贺丽娟.健胎液对小鼠子宫内膜LIF表达的影响.湖南中医药导报,2004;10(10):46-48.
[7] 田葱,王文辉,李娟,等.益肾种子丸对不明原因不孕症患者黄体中期子宫内膜E-钙黏素表达的影响.中医杂志,2006;47(11):834-835.
[8] 王冰洁,黄海红.补肾养血中药对无排卵性不孕的治疗作用及机理探讨.广州中医药大学学报,2002;19(3):189-191.
[9] 张树成,张志洲,刘效群等.补肾调经方药促进人着床期子宫内膜同步化的组织形态学观察.中国中医基础医学杂志,2002;8(4):48-49.
[10] 张树成,张志洲,刘彬,等.补肾调经方调经促排卵健内膜作用的临床实验研究.中医药学刊,2002;20(6):720-721.
[11] 张树成,沈明秀,吴志奎.补肾生血和补肾调经方药对老龄雌性金黄地鼠生殖器官组织形态的影响.中国民间疗法,1998;6(5):56-57.
[12] 陈秋梅,张树成,贺彬等.调经孕育方药对排卵障碍性不孕者子宫内膜感受态的影响—组织形态学分析.中医药学刊,2004;22(3):464-467.
[13] 刘艳娟,黄光英,陆付耳等.健胎液对胚泡着床障碍小鼠胚泡着床作用的研究.上海中医药杂志,2004;38(4):46-48.
[14] 龚萍,黄光英,刘艳娟等.健胎液改善胚泡着床障碍模型小鼠实验研究.山东中医杂志,2005;24(12):739-741.
[15] 黄冬梅,黄光英,陆付耳.补肾益气和血方中药对胚泡着床障碍小鼠子宫内膜表面胞饮突表达的影响.中华妇产科杂志,2004;39(4):230-233.
[16] 王素霞,孙玉英.安胎合剂对GnRH-a超排卵小鼠子宫内膜形态学的影响.中医药学报,2006;34(5):45-47.
[17] 张奕民.肾虚型不孕症和宫内灌注不良的相关性研究.江苏中医,1999;20(1):14-15.
[18] 张明敏,黄光英,陆付耳.补肾益气活血汤对多次助孕技术失败患者结局的影响[J] .微循环学杂志,2002;12(2):10-12.
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[20] 夏誉薇,张树成,蔡连香.女贞孕育汤对不孕症子宫内膜组织形态、雌孕激素受体和血管生成因子表达的影响.中国中医药信息杂志,2004;11(1):26-28.
[21] 夏誉薇,蔡连香,张树成.女贞孕育汤对鸡胚绒毛尿囊膜血管生成模型血清药理学研究.中医药学刊,2003;21(4):531-534.
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[23] 陈秋梅,张树成,沈明秀.调经孕育方药对排卵障碍性不孕者同步测试的卵泡和子宫内膜生长发育、血流特性的影响.中国实验方剂学杂志,2004;10(2):59-61.
[24] 张明敏,黄光英,陆付耳等.针刺对胚胎移植怀孕率的影响及其机理:随机安慰对照研究.中国针灸,2003;23(1):3-5.
[25] 张明敏,黄光英,陆付耳等.针刺对胚胎移植怀孕率的影响.中国针灸,2002;22(8):507-509.
[26] 黄冬梅,黄光英,陆付耳.补肾益气和血方对胚泡着床障碍小鼠子宫内膜腺体凋亡的影响.中国中西医结合杂志,2005;25(6)(基础理论研究特集):139-142.