干预“石门”穴对着床期小鼠子宫内膜容受性影响的实验研究
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摘要
目的:通过观察干预“石门”穴对着床期小鼠子宫内膜胞饮突、白血病抑制因子(LIF)表达及血清中孕酮(P)含量的影响,初步探讨干预“石门”穴对小鼠胚胎着床的影响及其作用机制、可能途径和相关靶点,为针灸疗法用于抗着床提供科学依据。
材料与方法:将昆明种雌性小鼠60只按体重分层随机分为空白对照组(A组)、电针“石门”组(B组)、手针“石门”组(C组)、埋线“石门”组(D组)、西药对照组(E组)、非穴电针组(F组),每组10只,本实验以妊娠第一天(Pd1)小鼠为研究对象。B、C、D组选取“石门”穴进行电针、手针、埋线刺激;E组皮下注射米非司酮;F组在非穴位点进行电针刺激。疗程:B、C、F组自Pd1上午8:00开始针刺,一天一次,连续5天;D组Pd1上午8:00施治一次;E组于Pd4上午9:00施治一次。以上各组小鼠均在Pd5上午9:30末次干预后处死,迅速剖腹取出子宫,用扫描电镜观察小鼠子宫内膜表面胞饮突的表达;光镜下观察小鼠子宫内膜形态变化(HE染色);用免疫组化SABC法检测小鼠子宫内膜LIF的表达;用放射免疫法检测小鼠血清中P的含量。
结果:
1.各组小鼠Pd5子宫内膜形态变化:A组小鼠腺腔内充盈着丰富的被染成淡红色的分泌物,腺腔扩大呈锯齿状,间质内血窦多。F组子宫内膜发育与A组相一致。B、C、D、E组小鼠子宫内膜比A组发育滞后。
2.各组小鼠Pd5子宫内膜表面胞饮突的表达:A组小鼠子宫内膜表面均匀分布着丰富的胞饮突,大小一致,胞饮突之间界限清楚,此即发育成熟的胞饮突;F组亦可见发育成熟的胞饮突;B、C、D组小鼠子宫内膜表面膜状突起明显低于A组,且大小不均,边界欠清,表面覆盖为微绒毛,子宫内膜表面胞饮突呈局灶性表达,且与胚胎发育不同步,胞饮突从形态上看较A组发育受到抑制,从发生时相上看延迟于A组;E组小鼠子宫内膜表面无胞饮突,可见呈多边形的上皮细胞,上皮细胞顶端膜覆以细的微绒毛,细胞间界限清楚,表现为增生早期的子宫内膜变化,其发育亦明显延后于A组。
3.各组小鼠Pd5血清中P含量结果和变化:B、C、D、E组小鼠血清中P含量与A组相比均有不同程度下降,比较差异显著(P＜0.05),有统计学意义。
4.各组小鼠Pd5子宫内膜LIF的表达:各组小鼠LIF表达部位均位于子宫内膜腔上皮和腺上皮细胞的胞浆;A、F组小鼠子宫内膜LIF的表达强度以“++”为主;B、C、D、E组小鼠子宫内膜LIF的表达强度以“+”为主。图像分析结果显示B、C、D、E组小鼠子宫内膜LIF的表达与A组比较差异显著(P＜0.05),有统计学意义。
结论:
1.干预“石门”穴在整体水平上对着床期小鼠有抗着床作用,进一步证实石门穴具有这一特异性;
2.干预“石门”穴影响小鼠子宫内膜容受性建立和着床窗开放,使子宫内膜和胚胎发育不同步,从而使得胚胎不能正常植入着床;
3.在抗着床过程中,干预“石门”穴的作用模式可能为“一点触发,多点调控,整合调节”,这种作用模式来源于孕酮的后续连锁反应;
4.电针、手针、埋线三种方法干预“石门”穴都能影响胚胎着床,其中以电针作用最明显,都是通过调节孕酮来实现的。
Purpose
To object the effects of intervening at "Shimen" on the expression of pinopodes and LIF on the mouse endometrium and progesterone during implantation period,so as to study the effect of intervening at "Shimen" on the implantation of mouse embryos and action mechanism and pathway and relative target and explain action character of acupuncture.
Material and methods
Sixty female mice were randomly divided into blank control group(group A,n=10),electro-acupuncture "Shimen" group(group B,n=10),hand-acupuncture "Shimen" group(group C,n=10),catgut embedding "Shimen" group(group D,n=10), positive medicine group(group E,n=10),and non-acupoint group(group F,n=10). Pregnance day 1 was determined on the vaginal plug day."Shimen" were treated with electroacupuncture and acupuncture;Catgut embedding group choose "Shimen";On Pd4,group E were injected subcutaneously with mifepristone diluted in propylene glycol;Group F were treated with electroacupuncture.Course of treatment:group B,C and D treated once on the first day,being treated 5 times insuccession;Group D treated one on first day;Group E infused with medicine on the fourth day.The animals used as experiment of every the above-mentioned groups put to death at 9:30 a.m on Pd5 after the end treatment.Progesterone was determined by radioimmunoassay;The morphologic structure of endometrium was observed by light microscope and scanning electron microscope;The expression of LIF was detected by immunohistochemistry techniques.
Results
1.The development of endometrium in group B,C,D and E was inhibited.
2.The expression of pinopodes in mouse endometrium.
In group A,day 5 of vaginal Plug,the endometrial surface is covered with fully developed pinopodes which are larger,completely smooth with well defined border of the cells,resembling a mushroom;group F is covered with fully developed pinopodes,too;In group B and C and D,day 5 of vaginal plug,the endometrial surface is covered with small projections without distinguished borders of cells.
The endometrial surface expresses developing pinopodes locally which are asynchronous;In group E,day 5 of vaginal plug,the endometrial surface is covered with hexagonal and flat appeal cells without any pinopodes formation.Compared with group A,the development of pinopodes lagged behind obviously and was repressed.
3.The resuIts and enchangings of the levels of P in the serums of different group mouses.
After treating,there are various decreases of comparing treat group(B,C,D and E) to group A(P＜0.05).
4.The expression of LIF in mouse endometrium of different group.
Immunostaining in mouse endometrial epithelial cell and glandular epithelial cell is observed on the implantation day with different level in different group.The results of Semi-quantitative assay show:compared with group A,the level of LIF is significantly decreased in groups of B,C,Dand E(P＜0.05).
Conclusion
1.Interventing at "Shimen" could inhibit blastocyst implantation in vivo. Specificity of acupoint was further confirmed;
2.Intervening at "Shimen" is not beneficial to establish endometrial respectively and to open the implantation window,thus affecting the establishment of embryo implantation;
3.In the circumstance of anti-implantation,intervention Shimen possesses the effect character of 'key point trigger-multidrop control-integrate regulation' This kind of character maybe derives from chain reaction after P is decreased;
4.Different needling methods have different effects on anti-implantation.Their mechanism may be by way of progesterone.
材料与方法:将昆明种雌性小鼠60只按体重分层随机分为空白对照组(A组)、电针“石门”组(B组)、手针“石门”组(C组)、埋线“石门”组(D组)、西药对照组(E组)、非穴电针组(F组),每组10只,本实验以妊娠第一天(Pd1)小鼠为研究对象。B、C、D组选取“石门”穴进行电针、手针、埋线刺激;E组皮下注射米非司酮;F组在非穴位点进行电针刺激。疗程:B、C、F组自Pd1上午8:00开始针刺,一天一次,连续5天;D组Pd1上午8:00施治一次;E组于Pd4上午9:00施治一次。以上各组小鼠均在Pd5上午9:30末次干预后处死,迅速剖腹取出子宫,用扫描电镜观察小鼠子宫内膜表面胞饮突的表达;光镜下观察小鼠子宫内膜形态变化(HE染色);用免疫组化SABC法检测小鼠子宫内膜LIF的表达;用放射免疫法检测小鼠血清中P的含量。
结果:
1.各组小鼠Pd5子宫内膜形态变化:A组小鼠腺腔内充盈着丰富的被染成淡红色的分泌物,腺腔扩大呈锯齿状,间质内血窦多。F组子宫内膜发育与A组相一致。B、C、D、E组小鼠子宫内膜比A组发育滞后。
2.各组小鼠Pd5子宫内膜表面胞饮突的表达:A组小鼠子宫内膜表面均匀分布着丰富的胞饮突,大小一致,胞饮突之间界限清楚,此即发育成熟的胞饮突;F组亦可见发育成熟的胞饮突;B、C、D组小鼠子宫内膜表面膜状突起明显低于A组,且大小不均,边界欠清,表面覆盖为微绒毛,子宫内膜表面胞饮突呈局灶性表达,且与胚胎发育不同步,胞饮突从形态上看较A组发育受到抑制,从发生时相上看延迟于A组;E组小鼠子宫内膜表面无胞饮突,可见呈多边形的上皮细胞,上皮细胞顶端膜覆以细的微绒毛,细胞间界限清楚,表现为增生早期的子宫内膜变化,其发育亦明显延后于A组。
3.各组小鼠Pd5血清中P含量结果和变化:B、C、D、E组小鼠血清中P含量与A组相比均有不同程度下降,比较差异显著(P＜0.05),有统计学意义。
4.各组小鼠Pd5子宫内膜LIF的表达:各组小鼠LIF表达部位均位于子宫内膜腔上皮和腺上皮细胞的胞浆;A、F组小鼠子宫内膜LIF的表达强度以“++”为主;B、C、D、E组小鼠子宫内膜LIF的表达强度以“+”为主。图像分析结果显示B、C、D、E组小鼠子宫内膜LIF的表达与A组比较差异显著(P＜0.05),有统计学意义。
结论:
1.干预“石门”穴在整体水平上对着床期小鼠有抗着床作用,进一步证实石门穴具有这一特异性;
2.干预“石门”穴影响小鼠子宫内膜容受性建立和着床窗开放,使子宫内膜和胚胎发育不同步,从而使得胚胎不能正常植入着床;
3.在抗着床过程中,干预“石门”穴的作用模式可能为“一点触发,多点调控,整合调节”,这种作用模式来源于孕酮的后续连锁反应;
4.电针、手针、埋线三种方法干预“石门”穴都能影响胚胎着床,其中以电针作用最明显,都是通过调节孕酮来实现的。
Purpose
To object the effects of intervening at "Shimen" on the expression of pinopodes and LIF on the mouse endometrium and progesterone during implantation period,so as to study the effect of intervening at "Shimen" on the implantation of mouse embryos and action mechanism and pathway and relative target and explain action character of acupuncture.
Material and methods
Sixty female mice were randomly divided into blank control group(group A,n=10),electro-acupuncture "Shimen" group(group B,n=10),hand-acupuncture "Shimen" group(group C,n=10),catgut embedding "Shimen" group(group D,n=10), positive medicine group(group E,n=10),and non-acupoint group(group F,n=10). Pregnance day 1 was determined on the vaginal plug day."Shimen" were treated with electroacupuncture and acupuncture;Catgut embedding group choose "Shimen";On Pd4,group E were injected subcutaneously with mifepristone diluted in propylene glycol;Group F were treated with electroacupuncture.Course of treatment:group B,C and D treated once on the first day,being treated 5 times insuccession;Group D treated one on first day;Group E infused with medicine on the fourth day.The animals used as experiment of every the above-mentioned groups put to death at 9:30 a.m on Pd5 after the end treatment.Progesterone was determined by radioimmunoassay;The morphologic structure of endometrium was observed by light microscope and scanning electron microscope;The expression of LIF was detected by immunohistochemistry techniques.
Results
1.The development of endometrium in group B,C,D and E was inhibited.
2.The expression of pinopodes in mouse endometrium.
In group A,day 5 of vaginal Plug,the endometrial surface is covered with fully developed pinopodes which are larger,completely smooth with well defined border of the cells,resembling a mushroom;group F is covered with fully developed pinopodes,too;In group B and C and D,day 5 of vaginal plug,the endometrial surface is covered with small projections without distinguished borders of cells.
The endometrial surface expresses developing pinopodes locally which are asynchronous;In group E,day 5 of vaginal plug,the endometrial surface is covered with hexagonal and flat appeal cells without any pinopodes formation.Compared with group A,the development of pinopodes lagged behind obviously and was repressed.
3.The resuIts and enchangings of the levels of P in the serums of different group mouses.
After treating,there are various decreases of comparing treat group(B,C,D and E) to group A(P＜0.05).
4.The expression of LIF in mouse endometrium of different group.
Immunostaining in mouse endometrial epithelial cell and glandular epithelial cell is observed on the implantation day with different level in different group.The results of Semi-quantitative assay show:compared with group A,the level of LIF is significantly decreased in groups of B,C,Dand E(P＜0.05).
Conclusion
1.Interventing at "Shimen" could inhibit blastocyst implantation in vivo. Specificity of acupoint was further confirmed;
2.Intervening at "Shimen" is not beneficial to establish endometrial respectively and to open the implantation window,thus affecting the establishment of embryo implantation;
3.In the circumstance of anti-implantation,intervention Shimen possesses the effect character of 'key point trigger-multidrop control-integrate regulation' This kind of character maybe derives from chain reaction after P is decreased;
4.Different needling methods have different effects on anti-implantation.Their mechanism may be by way of progesterone.
引文
[1]Jerome F,Strauss Ⅲ,Robert L,et al.Yen&Jaffe生殖内分泌[M].第5版.北京:人民卫生出版社,2006:324
[2]李沛,黄攸玉,王训立,等.针刺对雌性恒河猴生殖内分泌抑制性影响的实验研究[J].中国针灸,1998,8:486-488
[3]陈瑛,李晓文,耿军.人胚胎植入与激素-免疫-细胞因子及粘附分子网络系统[J].中华妇产科杂志,1999,34(6):382
[4]陈少宗,刘晶.从传统针灸学到现代针灸学-针灸现代化50年之回顾[J].医学与哲学,2006,27(9):57-59
[5]陈杰中,陈玉明.针灸石门-避孕[J].新中医药,1956,8(11):25
[6]赵柯.针灸石门节制生育32例[J].中国针灸,1991,1:20
[7]武子玉,陈藻琳.针灸石门避孕对子宫的影响[J].新中医药,1957,8(12):33
[8]张鸥,韩红.石门穴临床应用举隅[J].上海中医药杂志,2003,37(1):44-45
[9]李沛,黄秀兰,吴昌杰,等.针刺对雌性恒河猴月经周期雌二醇、睾酮分泌水平影响的实验研究[J].福建中医学院学报,1991,(2):18
[10]李沛,黄攸玉,徐文倩,等.针刺对女性生殖内分泌影响的实验研究[J].针刺研究,2002,4(27):277-280
[11]李沛,黄攸玉,王训立,等.针刺对雌性恒河猴生殖内分泌抑制性影响的实验研究[J].中国针灸,1998,8:486-488
[12]李沛,安晓英,徐文倩,等.针刺不同穴位对雌性恒河猴月经周期E2、T分泌的影响[J].福建中医学院学报,2000,10(2):19-22
[13]李沛,徐文倩,安晓英,等.不同时间针刺对雌性恒河猴生殖内分泌影响的实验研究[J].中国针灸,2002,22(3):189-192
[14]陈泽斌,柯晖,邹峰.大鼠脑缺血再灌注后不同时段中枢杏仁核CRF的表达变化[J].湖北中医学院学报,2006,8(3):5-6
[15]Jonathan S,Berk.Novak妇科学[M].第13版.北京:人民卫生出版社,2005:120
[1]杨甲三,曹一鸣.腧穴学[M].上海:上海科学技术出版社,1992:140
[2]李忠仁.实验针灸学[M].第2版.北京:中国中医药出版社,2007:255
[3]刘艳娟,黄光英,陆付耳,等.小鼠胚泡着床障碍模型的建立[J].中国药理学通报,2003,19(11):1315
[4]中村章.冲任新解[J].上海中医院杂志,1999,11:24-25
[5]Finn CA,Martin L.The control of implantation.J Reprod Fertil,1974,39:195-206
[6]Paria BC,Reese J,DasSK,et al.Deciphering the cross-talk of implantation:Advances and challenges.Science,2002,296:2185-2188
[7]Psychoyos A.Hormonal control of ovoinmplantation.Vitam Horm,1973,31:201
[8]Mclaren A.Blastocyst avtivation.In Segal SJ,Crozier R,Lorfman PA,Condliffe PG(eds).The Regulation of Mammalian Reprouction.London:Thomas Spsringfield,1973:321
[9]Wiliox AJ,Baird DD,Weiinberg CR.Time of implantation of the conceptus and loss of pregnancy.N Engl J Med,1999,340:1796-1799
[10]Jerome F,Strauss Ⅲ,Robert L,et al.Yen&Jaffe生殖内分泌[M].第5版.北京:人民卫生出版社,2006:324
[11]Psychoyos A,Mandon P.Study of the surface of the uterine epithelium by scanning electron microscope.Observations in.the rat at the 4th and 5th day of pregnancy.C R Acad Sci Hebd Seances Acad Sci D,1971,272:2723-2725
[12]Enders AC,Nelson DM.Pinocytotic activity of the uterus of the rat,Am J Anat,1973,138:277-299
[13]Nikas G.Endometrial recepticity:Changes in cell-surface morphology.Semin Reprod Med,2000,18:229
[14]NikasG,Aghajianova L.Endometrial pinopodes:somemore understanding on human implantation?.Reprod Biomed Online,2002,4[Supp13]:18-23
[15]Nikas G,Makrigiannakis A,Hovatta O,et al.Surface mor-phology of the human endometrium:basic and clinical aspects.Ann N Y hcad Sci,2000,900:316-324
[16]Nikas G.Uterine pinopodes as markers of uterine receptivityin clinical practice.Hun Reprod,1999,14(Supp12):99-106
[17]Bentin-Ley U,Sjogren A,Nilsson L,et al.Presence of uter-ine pinopodes at the embryo-endometrial interface during human implantationin vitro.Hum Reprod,1999,14(2)
[18]Nawroth F,LudwigM.What is the 'ideal' duration of progesterone supplementation before the transfer of cryopreserved-thawed embryos in estrogen/progesterone replacement protocols?[J].Hum Reprod,2005,20(5):1127-1134
[19]Bourgain C.Endometrial biopsy in the evaluation of endometrial receptivity [J].JGynecol Obstet BiolReprol(Paris),2004,33(l pt2):S13-17
[20]Younis JS,MatilskyM,RadinO,et al.Increased progesterone/estradiol ratio in the late follicular phase could be related to low ovarian reserve in invitro fertilization embryo transfer cycles with a long gonadotropin releasing horm one agonist protocol[J].Fertil Steri,2001,76(3):294-299
[21]罗国群,符爱珍,陈丽.GnRHa长方案促超排卵对胞饮突发育的影响[J].中国优生与遗传杂志,2007,15(6):103-105
[22]Aghajanova L,StavreusEversA,NikasY,et al.Coexpression of pinopodes and leukemia inhibitory factor,aswell as its receptor,in human endometrium[J].FertilSteri,2003,79(3)[Suppl1]:808-814
[23]NikasG,OsmanHD,JamesPT,et al.Endometrial pinopodes indicate a shift in the window of receptivity in IVF cycles[J]Hum Reprod,1999,14(3):787-792
[24]Stavreus-Evers A,Nikas G,Sahlin L,et al.Formation of pinopodes in human endometrium is associated with the concentration of progesterone and progesterone receptors.Fertil Steril,2001,76(4):782-791
[25]Jerome F.Strauss Ⅲ,Robert L.Barbieri.Yen&Jaffe生殖内分泌[M].第5版.北京:人民卫生出版社,2006:324
[26]祝彼得,司徒亮.中华妇产科学[M].第1版.北京:人民卫生出版社,1999:174-175
[27]董方莉,谭丽,郑英.种植窗期血清孕激素水平及子宫内膜整合素表达对IVF-ET结局的预测价值[J].生殖与避孕,2007,27(1):42-44
[28]谭宗建,李尚为,邱东升,等.大鼠血清孕激素水平对胚胎“植入窗”的预测作用[J].华西医大学报,2002,33(3):353-356
[29]Mlilligan SR,Finn CA.Minimal progesterone support required for the maintenance of pregnancy in mice.Hum Reprod,1997,12(3):602
[30]吴熙瑞.生殖内分泌学[M].第1版.北京:人民卫生出版社,1998:38-39
[31]Senturk LM,Arici A.Leukemia inhibitory factor in human reproduction.Am J Reprod Immu-nol,1998,39(2):144-151
[32]Laird SM,Tuckerman EM,Daoton CF,et al.The production of leukaemia inhibitory factor by human endometrium:presence in uterine flushing and production by cells in culture.Hum Reprod,1997,12(30):569
[33]BhattH,BrunetLJ,StewartCL.Uterine expression of leuKemia inhibitory facto concides with the onset of blastocyst implantation.Pro Natl Acad Sei USA,1991,88(24):11408-11412
[34]张炜,周剑萍,刘银坤,等.白血病抑制因子在小鼠围着床期子宫内膜表达[J].生殖医学杂志,2000,9:100-103
[35]Stewart CL,Kaspar P,Brunet LJ,et al.Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor.Nature,1992,359:76
[36]Vogiagis D,Marsh MM,Frey RC,et al.Leukemia inhibitory factor in human endomentrium throughout the menstrual cycle.J Endocrinol,1996,148(1):95-102.
[37]Chen DB,Hilsenrath R,Yang ZM,et al.Leukemia inhibitory factor in human endomentrium during the menstrual cycle:cellula(?) origin and action on production of glandular epithelial cell prostaglandin in vitro.Hum Reprod,1995,10(4):911-8
[38]Nachtigall MJ,Kliman HJ,Feinberg RF,et al.The effect of leukemia inhibitory factor(LIF)on trophoblast differentiation:a potential role in human implantation.J Clin Endorinol Metab,1996,81(2):801-806
[39]Sengupta J,Dhawan L&Ghosh D.Immunohistochemical localization of leukemia inhibitory factor,interleukins 1 and 6 at the primary implantation site in the rhesus monkey.Cytokine,2003,24(6):277-285
[40]吴焕淦,姚伟,周恩华,等.经穴特异性研究的思考[J].中国针灸,2007,27(1):59-62
[41]余安胜,赵英侠,严振国.足三里穴的显微结构[J].中国针灸,1999,19(1):27-28
[42]明彩荣,蔡朔,蔡玉文,等.电针“足三里”穴区深筋膜肥大细胞的荧光显微镜观察[J].针刺研究,2000,25(1):52
[43]赵宁侠,高巍,史恒军,等.电针足三里及非经非穴点对大鼠细胞免疫的影响[J].云南中医学院学报,2001,24(1):37-39
[44]马海峰,吴瑛,杨华元.穴位和非穴点刺激对大鼠骨骼肌能量代谢的影响[J].上海体育学院学报,2004,28(6):45-48
[45]张俊海,冯晓源,李霁,等.穴位和非穴位电针镇痛的脑功能磁共振对照研究[J].中国医学计算机成像杂志,2005,11(1):10-16
[46]游振铨,胡翔龙,张伟,等.以压迫阻滞针效定位的方法测试人体体表心包经的循行路线[J].中国针灸,1999,24(5):281-284
[47]许云祥,张家维,邓倩萍.穴位埋线疗法及其在癫痫治疗中的应用[J].中医药信息杂志[J],2003,20(1):35
[48]张燕华.针灸与免疫的新概念[J].华西医学,1998,13(1):38
[49]张燕华.经络-神经-内分泌-免疫网络[J].华西医学,1998,13(3):316-317
[50]骆永珍,张燕华,周荣幸.针灸与免疫[M].第1版.北京:人民卫生出社,2003:302-303
[51]王琳,史常旭,常青,等.围着床期子宫内膜表面形态超微结构的观察[J].中华妇产科杂志,2001,36(8):500
[52]Stavreus Evers A,Nikas G,Sahlin L,et al.Formation of Pinop-odes in Human Endometrium is Associated with the Concentration of Progesterone and Progesterone receptors[J].Fertil Steril,2001,76(4):782-791
[53]Piccinni MP,Scaletti C,Maggi E,et al,Role of hormone-controlled Th1-and Th2-type cytokines in successful pregnancy.J Neuroimmunol,2000,109(1):30-33
[2]李沛,黄攸玉,王训立,等.针刺对雌性恒河猴生殖内分泌抑制性影响的实验研究[J].中国针灸,1998,8:486-488
[3]陈瑛,李晓文,耿军.人胚胎植入与激素-免疫-细胞因子及粘附分子网络系统[J].中华妇产科杂志,1999,34(6):382
[4]陈少宗,刘晶.从传统针灸学到现代针灸学-针灸现代化50年之回顾[J].医学与哲学,2006,27(9):57-59
[5]陈杰中,陈玉明.针灸石门-避孕[J].新中医药,1956,8(11):25
[6]赵柯.针灸石门节制生育32例[J].中国针灸,1991,1:20
[7]武子玉,陈藻琳.针灸石门避孕对子宫的影响[J].新中医药,1957,8(12):33
[8]张鸥,韩红.石门穴临床应用举隅[J].上海中医药杂志,2003,37(1):44-45
[9]李沛,黄秀兰,吴昌杰,等.针刺对雌性恒河猴月经周期雌二醇、睾酮分泌水平影响的实验研究[J].福建中医学院学报,1991,(2):18
[10]李沛,黄攸玉,徐文倩,等.针刺对女性生殖内分泌影响的实验研究[J].针刺研究,2002,4(27):277-280
[11]李沛,黄攸玉,王训立,等.针刺对雌性恒河猴生殖内分泌抑制性影响的实验研究[J].中国针灸,1998,8:486-488
[12]李沛,安晓英,徐文倩,等.针刺不同穴位对雌性恒河猴月经周期E2、T分泌的影响[J].福建中医学院学报,2000,10(2):19-22
[13]李沛,徐文倩,安晓英,等.不同时间针刺对雌性恒河猴生殖内分泌影响的实验研究[J].中国针灸,2002,22(3):189-192
[14]陈泽斌,柯晖,邹峰.大鼠脑缺血再灌注后不同时段中枢杏仁核CRF的表达变化[J].湖北中医学院学报,2006,8(3):5-6
[15]Jonathan S,Berk.Novak妇科学[M].第13版.北京:人民卫生出版社,2005:120
[1]杨甲三,曹一鸣.腧穴学[M].上海:上海科学技术出版社,1992:140
[2]李忠仁.实验针灸学[M].第2版.北京:中国中医药出版社,2007:255
[3]刘艳娟,黄光英,陆付耳,等.小鼠胚泡着床障碍模型的建立[J].中国药理学通报,2003,19(11):1315
[4]中村章.冲任新解[J].上海中医院杂志,1999,11:24-25
[5]Finn CA,Martin L.The control of implantation.J Reprod Fertil,1974,39:195-206
[6]Paria BC,Reese J,DasSK,et al.Deciphering the cross-talk of implantation:Advances and challenges.Science,2002,296:2185-2188
[7]Psychoyos A.Hormonal control of ovoinmplantation.Vitam Horm,1973,31:201
[8]Mclaren A.Blastocyst avtivation.In Segal SJ,Crozier R,Lorfman PA,Condliffe PG(eds).The Regulation of Mammalian Reprouction.London:Thomas Spsringfield,1973:321
[9]Wiliox AJ,Baird DD,Weiinberg CR.Time of implantation of the conceptus and loss of pregnancy.N Engl J Med,1999,340:1796-1799
[10]Jerome F,Strauss Ⅲ,Robert L,et al.Yen&Jaffe生殖内分泌[M].第5版.北京:人民卫生出版社,2006:324
[11]Psychoyos A,Mandon P.Study of the surface of the uterine epithelium by scanning electron microscope.Observations in.the rat at the 4th and 5th day of pregnancy.C R Acad Sci Hebd Seances Acad Sci D,1971,272:2723-2725
[12]Enders AC,Nelson DM.Pinocytotic activity of the uterus of the rat,Am J Anat,1973,138:277-299
[13]Nikas G.Endometrial recepticity:Changes in cell-surface morphology.Semin Reprod Med,2000,18:229
[14]NikasG,Aghajianova L.Endometrial pinopodes:somemore understanding on human implantation?.Reprod Biomed Online,2002,4[Supp13]:18-23
[15]Nikas G,Makrigiannakis A,Hovatta O,et al.Surface mor-phology of the human endometrium:basic and clinical aspects.Ann N Y hcad Sci,2000,900:316-324
[16]Nikas G.Uterine pinopodes as markers of uterine receptivityin clinical practice.Hun Reprod,1999,14(Supp12):99-106
[17]Bentin-Ley U,Sjogren A,Nilsson L,et al.Presence of uter-ine pinopodes at the embryo-endometrial interface during human implantationin vitro.Hum Reprod,1999,14(2)
[18]Nawroth F,LudwigM.What is the 'ideal' duration of progesterone supplementation before the transfer of cryopreserved-thawed embryos in estrogen/progesterone replacement protocols?[J].Hum Reprod,2005,20(5):1127-1134
[19]Bourgain C.Endometrial biopsy in the evaluation of endometrial receptivity [J].JGynecol Obstet BiolReprol(Paris),2004,33(l pt2):S13-17
[20]Younis JS,MatilskyM,RadinO,et al.Increased progesterone/estradiol ratio in the late follicular phase could be related to low ovarian reserve in invitro fertilization embryo transfer cycles with a long gonadotropin releasing horm one agonist protocol[J].Fertil Steri,2001,76(3):294-299
[21]罗国群,符爱珍,陈丽.GnRHa长方案促超排卵对胞饮突发育的影响[J].中国优生与遗传杂志,2007,15(6):103-105
[22]Aghajanova L,StavreusEversA,NikasY,et al.Coexpression of pinopodes and leukemia inhibitory factor,aswell as its receptor,in human endometrium[J].FertilSteri,2003,79(3)[Suppl1]:808-814
[23]NikasG,OsmanHD,JamesPT,et al.Endometrial pinopodes indicate a shift in the window of receptivity in IVF cycles[J]Hum Reprod,1999,14(3):787-792
[24]Stavreus-Evers A,Nikas G,Sahlin L,et al.Formation of pinopodes in human endometrium is associated with the concentration of progesterone and progesterone receptors.Fertil Steril,2001,76(4):782-791
[25]Jerome F.Strauss Ⅲ,Robert L.Barbieri.Yen&Jaffe生殖内分泌[M].第5版.北京:人民卫生出版社,2006:324
[26]祝彼得,司徒亮.中华妇产科学[M].第1版.北京:人民卫生出版社,1999:174-175
[27]董方莉,谭丽,郑英.种植窗期血清孕激素水平及子宫内膜整合素表达对IVF-ET结局的预测价值[J].生殖与避孕,2007,27(1):42-44
[28]谭宗建,李尚为,邱东升,等.大鼠血清孕激素水平对胚胎“植入窗”的预测作用[J].华西医大学报,2002,33(3):353-356
[29]Mlilligan SR,Finn CA.Minimal progesterone support required for the maintenance of pregnancy in mice.Hum Reprod,1997,12(3):602
[30]吴熙瑞.生殖内分泌学[M].第1版.北京:人民卫生出版社,1998:38-39
[31]Senturk LM,Arici A.Leukemia inhibitory factor in human reproduction.Am J Reprod Immu-nol,1998,39(2):144-151
[32]Laird SM,Tuckerman EM,Daoton CF,et al.The production of leukaemia inhibitory factor by human endometrium:presence in uterine flushing and production by cells in culture.Hum Reprod,1997,12(30):569
[33]BhattH,BrunetLJ,StewartCL.Uterine expression of leuKemia inhibitory facto concides with the onset of blastocyst implantation.Pro Natl Acad Sei USA,1991,88(24):11408-11412
[34]张炜,周剑萍,刘银坤,等.白血病抑制因子在小鼠围着床期子宫内膜表达[J].生殖医学杂志,2000,9:100-103
[35]Stewart CL,Kaspar P,Brunet LJ,et al.Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor.Nature,1992,359:76
[36]Vogiagis D,Marsh MM,Frey RC,et al.Leukemia inhibitory factor in human endomentrium throughout the menstrual cycle.J Endocrinol,1996,148(1):95-102.
[37]Chen DB,Hilsenrath R,Yang ZM,et al.Leukemia inhibitory factor in human endomentrium during the menstrual cycle:cellula(?) origin and action on production of glandular epithelial cell prostaglandin in vitro.Hum Reprod,1995,10(4):911-8
[38]Nachtigall MJ,Kliman HJ,Feinberg RF,et al.The effect of leukemia inhibitory factor(LIF)on trophoblast differentiation:a potential role in human implantation.J Clin Endorinol Metab,1996,81(2):801-806
[39]Sengupta J,Dhawan L&Ghosh D.Immunohistochemical localization of leukemia inhibitory factor,interleukins 1 and 6 at the primary implantation site in the rhesus monkey.Cytokine,2003,24(6):277-285
[40]吴焕淦,姚伟,周恩华,等.经穴特异性研究的思考[J].中国针灸,2007,27(1):59-62
[41]余安胜,赵英侠,严振国.足三里穴的显微结构[J].中国针灸,1999,19(1):27-28
[42]明彩荣,蔡朔,蔡玉文,等.电针“足三里”穴区深筋膜肥大细胞的荧光显微镜观察[J].针刺研究,2000,25(1):52
[43]赵宁侠,高巍,史恒军,等.电针足三里及非经非穴点对大鼠细胞免疫的影响[J].云南中医学院学报,2001,24(1):37-39
[44]马海峰,吴瑛,杨华元.穴位和非穴点刺激对大鼠骨骼肌能量代谢的影响[J].上海体育学院学报,2004,28(6):45-48
[45]张俊海,冯晓源,李霁,等.穴位和非穴位电针镇痛的脑功能磁共振对照研究[J].中国医学计算机成像杂志,2005,11(1):10-16
[46]游振铨,胡翔龙,张伟,等.以压迫阻滞针效定位的方法测试人体体表心包经的循行路线[J].中国针灸,1999,24(5):281-284
[47]许云祥,张家维,邓倩萍.穴位埋线疗法及其在癫痫治疗中的应用[J].中医药信息杂志[J],2003,20(1):35
[48]张燕华.针灸与免疫的新概念[J].华西医学,1998,13(1):38
[49]张燕华.经络-神经-内分泌-免疫网络[J].华西医学,1998,13(3):316-317
[50]骆永珍,张燕华,周荣幸.针灸与免疫[M].第1版.北京:人民卫生出社,2003:302-303
[51]王琳,史常旭,常青,等.围着床期子宫内膜表面形态超微结构的观察[J].中华妇产科杂志,2001,36(8):500
[52]Stavreus Evers A,Nikas G,Sahlin L,et al.Formation of Pinop-odes in Human Endometrium is Associated with the Concentration of Progesterone and Progesterone receptors[J].Fertil Steril,2001,76(4):782-791
[53]Piccinni MP,Scaletti C,Maggi E,et al,Role of hormone-controlled Th1-and Th2-type cytokines in successful pregnancy.J Neuroimmunol,2000,109(1):30-33