三种中药有效成分对小鼠2-细胞胚胎体外培养抗氧化作用及冷冻胚胎移植效果研究
|
摘要
本研究主要探讨mCZB分别添加黄芩苷(Baicalin,Bai)、川芎嗪(Ligustrazine,Lig)、小檗碱(Berberine,BR)对小鼠2-细胞胚胎体外发育抗氧化作用的影响,并比较各组胚胎冷冻解冻效果及冷冻胚胎移植妊娠率和产仔率等,以进一步探讨利用三种中药有效成分改善小鼠早期胚胎体外培养微环境、提高体外培养胚胎质量和移植胚胎的发育潜力,为更好地改进哺乳动物胚胎培养液成分和培养条件提供可借鉴的资料和途径。
试验一:以mCZB添加抗生素为对照组,分别添加4μg/mlBai、0.5μg/mlLig和0.1μg/mlBR为试验组,比较各组胚胎孵化率、微滴中一氧化氮(Nitric oxide,NO)和丙二醛(malondialdehyde,MDA)含量及孵化胚胎的细胞数量。结果:各试验组胚胎孵化率,Bai组(80.2%,81.3%)、Lig组(78.9%,77.6%)及BR组(76.7%,71.6%)极显著高于对照组(50.7%,47.8%)(P<0.01)。各组间NO含量,72 h较24 h均有减少,而120 h又均有增加且最高。各组间MDA含量,72 h和120 h与24 h比较呈现增加趋势,除120hBai组,对照组均高于试验组。孵化胚胎细胞计数结果显示,BR组、Lig组和Bai组(87.2±8.6,83.9±7.7,81.9±6.2)与对照组(77.4±5.6)差异极显著(P<0.01)。表明,三种中药有效成分均能显著促进小鼠早期胚胎体外发育及胚胎细胞增殖,并对NO和MDA含量有一定影响。
试验二:以试验一为基础,2-细胞胚胎体外培养至桑椹、囊胚期进行常规冷冻,比较各组胚胎冷冻解冻效果及两种程序对胚胎冷冻解冻的影响。结果:冷冻解冻桑椹胚体外培养8～14 h囊胚发育率,程序一中Bai组(65.8%)显著高于对照组(48.6%)和BR组(46.7%)(P<0.05),Lig组(81.1%)极显著高于对照组(48.6%)和BR组(46.7%)(P<0.01);程序二中Bai组(80.6%)、Lig组(78.3%)极显著高于对照组(55.2%)和BR组(47.6%)(P<0.01)。冷冻解冻囊胚体外培养6～8 h囊胚发育率,程序一中各组间无显著差异(P>0.05);程序二中,中药有效成分各组间差异不显著(P>0.05),但均显著高于对照组(61.2%)(P<0.05)。两种冷冻程序冷冻效果差异不显著(P>0.05)。表明,添加Bai、Lig对提高胚胎体外发育质量效果明显,显著提高了冷冻胚胎发育率。
试验三:以试验二为基础,将各组冷冻解冻胚胎培养发育至囊胚期进行子宫移植。结果:移植妊娠率以Bai组(56.3%)、Lig组(52.6%)和BR组(55.0%)均高于对照组(46.2%);产仔率以Bai组(63.3%)、Lig组(67.3%)和BR组(60.5%),均高于对照组(52.8%),但差异不显著(P>0.05)。
综上结果表明:三种中药有效成份Bai、Lig、BR能显著提高小鼠2-细胞胚胎体外发育质量,并有利于提高冷冻解冻后移植胚胎的发育潜力。
This study was mainly approached the antioxidation effect of adding Baicalin(Bai), Ligustrazine(Lig)and Berberine(BR)in mCZB respectively on 2-cell mouse embryos in vitro culture ,and comparedthe effect of froze-thaw, pregnancy rate and litter rate after embryos transfer etc. in order to furtherinvestigate the value of these three Chinese Medicine effective constituents on improving themicro-environment of preimplantation embryos in vitro culture on mouse and enhancing the embryoquality and the developmental potential of embryos after embryos transfer , which can provide valuabledata and access to enhance the mammalian embryo culture medium composition and culture condition.
The group added antibiotics to mCZB was regarded as the control, while the others supplemented with0.1μg/ml BR, 0.5μg/ml Lig and 4μg/ml Bai respectively as experiment groups in the first experiment tocompare the rates of hatched embryos, the contents of nitric oxide(NO) and malondialdehyde(MDA) in themicrodrop, and the cell amount of hatched embryos. As a result, the rates of hatched embryos from thethree groups of Bai ( 80.2% , 81.3% ), Lig (78.9%,77.6% )and BR (76.7% ,71.6%) was significantly higher(P<0.01) than control group (50.7%、47.8%) . The content of NO decreased at 72h as compared with that of24h in every experiment group, but it increased to the highest point at 120h. As a whole, raise trend wasevident for the content of MDA in every group at 72h and 120h, and the content of MDA in the control washigher than that in experiment groups at 120h except for Bai group. According to the cell amount ofhatched embryos, the difference between experiment groups(BR(87.2±8.6), Lig(83.9±7.7) ,Bai(81.9±6.2))and control group(77.4±5.6)were extremely obvious (P<0.01). The result showed that these three kinds ofChinese Medicine effective constituents could efficiently promote early embryos development and increasethe cell amount of hatched embryo, and the contents of NO and MDA were affected as well in vitro cultureon mouse.
Based on the first experiment, the second one in which each group were conserved throughconventional freezing process from 2-cell embryo in vitro culture to the stage of morula and blastocyst wasoriented to compare the effects of froze-thaw in each group and the difference between the two methods.The results of morula and blastocyst showed that in the first procedure the development rate of blastocystcultured for 8～14 hours after froze-thaw in the Bai(65.8%)was superior(P<0.05)to that of the control(48.6%)and the BR(46.7%), and the Lig(81.1%)was significantly higher (P<0.01) than that ofthe control and the BR(46.7%). In the second procedure, the Ba(i80.6%)was significantly higher (P<0.01)than the control(55.2%)and the BR(47.6%)in vitro culture. Through analysis of the development rate ofblastocyst cultured for 6～8 hours in the first procedure, there was no significant difference(P>0.05)ineach group. In the second procedure, there was no significant difference(P>0.05)in each group, but theeffect of Chinese Medicine was superior(P<0.05)to that of control group(61.2%)in vitro culture. Theresults indicated there was no significant difference(P>0.05)in the different procedures. Bai and Lig cansignificantly enhance the development quality of embryos and the development rate of freezing embryos invitro culture.
Based on Experiment two, froze-thaw embryos were cultured to the stage of blastula and thentransferred to uterus in experiment three. As a result, the pregnancy rate from the three groups of Bai(56.3% ),Lig (52.6% )and BR (55.0%) was higher than that of control group (46.2%),but there was notobvious difference between experiment groups and control group;the birth rate from the three groups ofBai (63.3% ), Lig(67.3% )and BR (60.5%) was higher than control group (52.8%),but there was notsignificant difference either.
These results indicated the three Chinese Medicine effective constituents(Bai, Lig , BR)can distinctlyimprove the quality of 2-cell mouse embryo in vitro culture and be propitious to the developmentalpotential of embryos following embryos transfer after froze-thaw embryos.
试验一:以mCZB添加抗生素为对照组,分别添加4μg/mlBai、0.5μg/mlLig和0.1μg/mlBR为试验组,比较各组胚胎孵化率、微滴中一氧化氮(Nitric oxide,NO)和丙二醛(malondialdehyde,MDA)含量及孵化胚胎的细胞数量。结果:各试验组胚胎孵化率,Bai组(80.2%,81.3%)、Lig组(78.9%,77.6%)及BR组(76.7%,71.6%)极显著高于对照组(50.7%,47.8%)(P<0.01)。各组间NO含量,72 h较24 h均有减少,而120 h又均有增加且最高。各组间MDA含量,72 h和120 h与24 h比较呈现增加趋势,除120hBai组,对照组均高于试验组。孵化胚胎细胞计数结果显示,BR组、Lig组和Bai组(87.2±8.6,83.9±7.7,81.9±6.2)与对照组(77.4±5.6)差异极显著(P<0.01)。表明,三种中药有效成分均能显著促进小鼠早期胚胎体外发育及胚胎细胞增殖,并对NO和MDA含量有一定影响。
试验二:以试验一为基础,2-细胞胚胎体外培养至桑椹、囊胚期进行常规冷冻,比较各组胚胎冷冻解冻效果及两种程序对胚胎冷冻解冻的影响。结果:冷冻解冻桑椹胚体外培养8～14 h囊胚发育率,程序一中Bai组(65.8%)显著高于对照组(48.6%)和BR组(46.7%)(P<0.05),Lig组(81.1%)极显著高于对照组(48.6%)和BR组(46.7%)(P<0.01);程序二中Bai组(80.6%)、Lig组(78.3%)极显著高于对照组(55.2%)和BR组(47.6%)(P<0.01)。冷冻解冻囊胚体外培养6～8 h囊胚发育率,程序一中各组间无显著差异(P>0.05);程序二中,中药有效成分各组间差异不显著(P>0.05),但均显著高于对照组(61.2%)(P<0.05)。两种冷冻程序冷冻效果差异不显著(P>0.05)。表明,添加Bai、Lig对提高胚胎体外发育质量效果明显,显著提高了冷冻胚胎发育率。
试验三:以试验二为基础,将各组冷冻解冻胚胎培养发育至囊胚期进行子宫移植。结果:移植妊娠率以Bai组(56.3%)、Lig组(52.6%)和BR组(55.0%)均高于对照组(46.2%);产仔率以Bai组(63.3%)、Lig组(67.3%)和BR组(60.5%),均高于对照组(52.8%),但差异不显著(P>0.05)。
综上结果表明:三种中药有效成份Bai、Lig、BR能显著提高小鼠2-细胞胚胎体外发育质量,并有利于提高冷冻解冻后移植胚胎的发育潜力。
This study was mainly approached the antioxidation effect of adding Baicalin(Bai), Ligustrazine(Lig)and Berberine(BR)in mCZB respectively on 2-cell mouse embryos in vitro culture ,and comparedthe effect of froze-thaw, pregnancy rate and litter rate after embryos transfer etc. in order to furtherinvestigate the value of these three Chinese Medicine effective constituents on improving themicro-environment of preimplantation embryos in vitro culture on mouse and enhancing the embryoquality and the developmental potential of embryos after embryos transfer , which can provide valuabledata and access to enhance the mammalian embryo culture medium composition and culture condition.
The group added antibiotics to mCZB was regarded as the control, while the others supplemented with0.1μg/ml BR, 0.5μg/ml Lig and 4μg/ml Bai respectively as experiment groups in the first experiment tocompare the rates of hatched embryos, the contents of nitric oxide(NO) and malondialdehyde(MDA) in themicrodrop, and the cell amount of hatched embryos. As a result, the rates of hatched embryos from thethree groups of Bai ( 80.2% , 81.3% ), Lig (78.9%,77.6% )and BR (76.7% ,71.6%) was significantly higher(P<0.01) than control group (50.7%、47.8%) . The content of NO decreased at 72h as compared with that of24h in every experiment group, but it increased to the highest point at 120h. As a whole, raise trend wasevident for the content of MDA in every group at 72h and 120h, and the content of MDA in the control washigher than that in experiment groups at 120h except for Bai group. According to the cell amount ofhatched embryos, the difference between experiment groups(BR(87.2±8.6), Lig(83.9±7.7) ,Bai(81.9±6.2))and control group(77.4±5.6)were extremely obvious (P<0.01). The result showed that these three kinds ofChinese Medicine effective constituents could efficiently promote early embryos development and increasethe cell amount of hatched embryo, and the contents of NO and MDA were affected as well in vitro cultureon mouse.
Based on the first experiment, the second one in which each group were conserved throughconventional freezing process from 2-cell embryo in vitro culture to the stage of morula and blastocyst wasoriented to compare the effects of froze-thaw in each group and the difference between the two methods.The results of morula and blastocyst showed that in the first procedure the development rate of blastocystcultured for 8～14 hours after froze-thaw in the Bai(65.8%)was superior(P<0.05)to that of the control(48.6%)and the BR(46.7%), and the Lig(81.1%)was significantly higher (P<0.01) than that ofthe control and the BR(46.7%). In the second procedure, the Ba(i80.6%)was significantly higher (P<0.01)than the control(55.2%)and the BR(47.6%)in vitro culture. Through analysis of the development rate ofblastocyst cultured for 6～8 hours in the first procedure, there was no significant difference(P>0.05)ineach group. In the second procedure, there was no significant difference(P>0.05)in each group, but theeffect of Chinese Medicine was superior(P<0.05)to that of control group(61.2%)in vitro culture. Theresults indicated there was no significant difference(P>0.05)in the different procedures. Bai and Lig cansignificantly enhance the development quality of embryos and the development rate of freezing embryos invitro culture.
Based on Experiment two, froze-thaw embryos were cultured to the stage of blastula and thentransferred to uterus in experiment three. As a result, the pregnancy rate from the three groups of Bai(56.3% ),Lig (52.6% )and BR (55.0%) was higher than that of control group (46.2%),but there was notobvious difference between experiment groups and control group;the birth rate from the three groups ofBai (63.3% ), Lig(67.3% )and BR (60.5%) was higher than control group (52.8%),but there was notsignificant difference either.
These results indicated the three Chinese Medicine effective constituents(Bai, Lig , BR)can distinctlyimprove the quality of 2-cell mouse embryo in vitro culture and be propitious to the developmentalpotential of embryos following embryos transfer after froze-thaw embryos.
引文
[1]郑荣梁,黄中洋.自由基医学与农学基础[M].北京:高等教育出版社&施普林格出版社,2001:1～27
[2]赵保路.氧自由基和天然抗氧化剂[M].北京:科学技术出版社,1999
[3]刘建国,王宪,陈明哲.一氧化氮和细胞因子之间的相互调节作用[J].生理科学进展,2000,31(1):61～64
[4]王雁.一氧化氮对女性生殖系统的影响[ J].首都医药,2003,10(4):44～45
[5]Rossellli M,Keller PJ,Dubey PK.Role of nitric oxide in biology,physiology andpathophysiology of reproduction[J].Hum Reprod Up date,1998,4(1):3～24
[6]郑荣梁,张红.自由基对人精子功能的调控[J].扬州大学学报(自然科学版),1999,35(3):135～138
[7]Dey SK,Komura F,Mukherjee A,et al.Cyclic AMP and Cyclic GMP in rabbitblastocysts[J].ReProd Fertil,1978,52(2):235～237
[8]王元兴,郎介金.动物繁殖学[M].南京:江苏科学技术出版社,1993,166～174
[9]Brief S,Chew BP.effect of vitamin A and β-carotene on reproductive performance ingilts[J].J Anim Sci,1985,60(4):998～1004
[10]Quinn P,Harlow GM.The effect of oxygen on the development of preimplantationmouse embryos in vitro[J].J Exp Zool,1978,206(1):73～80
[11]Flood MR,Wiebold JL.Glucose metabolsm by preimplantation pig embryos[J],JReprod Fert,1988,84(1):7～12
[12]Laloraya M,Kumar GP,Laloraya MM.A possible role of superoxide anion radicalin the process of blastocyst implantation in musculus.Biochem Biophys Res Commun,1989,161(2):762～770
[13]Nasr-Esfahani MH,Aitken JR,Johnson MH.hydrogen peroxide levels in mouseoocytes and early cleavage stage embryos developed in vitro or in vivo[J].Development,1990,109(2):501～507
[14]Noda Y,Watsumoto H,Umadka Y,et al.Involvement of superoxide radicals in themouse two-cell block[J].Mol Reprod Dev.1991,28(4):356～360
[15]Li JM,Foote RH,Simkin M.Development of rabbit zygotes cultured in protein-freemedium with catalase,taurine or superoxide dismutase[J].Biol Reprod,1993,49(1):33～37
[16]Payne SR,Munday R,Thompson JG.Addition of superoxide dismutase and catalasedoes not necessarily overcome developmental retardation of one-cell mouse embryo duringin-vitro culture[J].Reprod Fertil Dev,1992,4(2):167～174
[17]Thomas M,Jain S,Kumar GP,et al.A programmed oxyradical burst causes hatchingof mouse blastocysts[J].Journal of cell science,1997,110(14):1597～1602
[18]Kuo RC,Baxter GT,Thompson SH,et al.NO is necessary and sufficient for eggactivation at fertilization[J].Nature,2000,406(6796):633～636
[19]Novaro V,Gonzalez E,Rettori V,et al.Nitric oxide synthase regulation duringembryonic implantation[J].Reprod Fertil Dev,1997,9(5):557～564
[20]Duran-Reyes G,Gomez-Melendez MR,Brena G.Nitric oxide Synthase inhibitionsuppresses implantation and decreases cGMP concentration and protein peroxidation[J].LifeScience,1999,65(21):2259～2268
[21]Gouge RC,Marshburn P,Gordon BE,et al.Nitric oxide as a regulator embryonicdevelopment[J].Biol Reprod,1998,58(4):875～879
[22]Abe K,Matsuoka K,Inoue N,et al.Messenger RNA of neuronal nitric oxidesynthase is expressed and possibly functions in mouse oocytes and embryos duringpreimplantation development[J].Biomed Res,1999,20(1):61～65
[23]Athanassakis I,Aifantis I,Baritakis S,et al.Nitric Oxide production bypre-implantation embryo in response to embryotoxic factors.Cellular Physiology andBiochemistry,2000,10(3):169～176
[24]解启发,邓泽沛.NO 在小老鼠早期胚胎吸收过程中的作用[J].中国兽医学报,2002,22(1):90～93
[25]周思畅,周剑涛.一氧化氮介导细胞凋亡的分子基础[J].生命科学,2002,14(3):135～138
[26]Purcell TL,Given R,Chwalisz K,et al.Nitric oxide synthase distribution duringimplantation in the mouse[J].Mol Hum Reprod,1999,5(5):467～475
[27]Farina M,Ribeiro ML,Ogando D,et al.IL-1 alpha augments prostaglandinsynthesis in pregnant rat uteriby a nitric oxide mediated mechanism[J].Prostaglandins LeukotEssent Fatty Acids ,2000,62(4):243～247
[28]王建辰,章孝荣.动物生殖调控[M].安徽:安徽科学技术出版社.1998:102
[29]Biswas S,Kabir SN.Therole of Nitric oxide in the process of implantation inrats[J].Journal of Reproduction and Fertility,1998,114(1):157～161
[30]朱猛进,丁家酮.一氧化氮(NO)对动物妊娠生理的调节[J].贵州畜牧兽医,2000,24(1):10～11
[31]林毅宏.自由基与中药 BRM 效应[J].福建中医药,1992,(4):41
[32]Robak J . Flavonids are scavengers of super oxideanions Biochemical [J].Pharmacology,1988,37:837
[33]Husain SR.Hydroxyl radical scavenging activity of flavonoids[J].Phytochemistry,1987,26(9):2489
[34]吴文林,胡天喜.几种黄酮类化合物对羟自由基引起的损伤的保护作用.自由基生命科学进展(第 5 集)[M].北京:原子能出版社,1997,101
[35]康鑫,方允中,刘学英.茶多酚清除超氧化物自由基的效能及其对肿瘤细胞 DNA生物合成的影响.自由基生命科学进展(第 1 集)[M].北京:原子能出版社,1993,163
[36]宋洪涛,郭涛,宓鹤鸣.天然药物中的抗氧化剂[J].中草药,1991,22(7):331
[37]詹皓,沈竟民,姜桂荣.中药抗氧化作用的研究概况[J].中国中药杂志,1990,15(10):54
[38]姚文兵,高向东,周慧萍,等.大黄的生化学研究 XXXVI 波叶大黄多糖抗衰老作用的实验研究[J].中国药科大学学报,1991,22(3):167
[39]劳汉昌,张宝华,胡侦明.脊髓损伤早期三七总皂苷抗氧化作用的实验研究[J].云南医药,1995,16(5):355
[40]何凤慈,唐汝愚,姚丹帆.粉防已碱对急性炎症血管通透性和嗜中性白细胞功能的影响[J].中国药理学报,1989,10(3):249.
[41]王利津,徐强.黄连解毒汤的抗氧化作用研究[J].中国药科大学学报,2001,32(1):51～53
[42]龚跃新,孙云,林安平.补气法、补血法抗自由基操作的比较研究[J].中国中药杂志,1993,18(7):438
[43]张其兰,孙志福.复方何首乌抗氧化作用的研究[J].自由基生命科学进展(第 1集)[M].北京:原子能出版社,1993:189
[44]Guerin P,Mouatassim S,Menezo Y.Oxidative stress and protection against reactiveoxygen species in the preimplantation embryo and its surroundings[J].HumReprod Update,2001,7:175～189
[45]Noda Y,Goto Y,Umaoka Y,et al.Culture of Human Embryos in AlphaModification of Eagle's Mdium under Low Oxygen Tensionan Low Illumination[J].FertilSteril,1994,62:1022～1027
[46]Umaoka Y,Noda Y,Narimoto K,et al.Developmental Potentiality Of EmbryosCultured under Low Oxygen Tension with Superoxide Dismutase[J].J In Vitro Fert EmbryoTransf,1991,8(5):245～249
[47]Herrick JB.Injectable β-carotene Appears to Improve Reproduction[J].Feed stuff,1993,7:13～17
[48]吴师光,董孝国.一种古老的生物信息分子-退黑素[J].商丘师范学院学报,2003,19(2):102～103
[49]关钧.环核苷酸与中药效功能综述[J].浙江中医杂志,1989,244(5):234～236
[50]孙忠亲.中草药对环核苷酸影响的研究进展[J].中草药,1990,21(12):31～32
[51]阮娜,宋晓平.补益药对体外培养细胞的影响[A].中兽医药研究进展,2003,7:158～160
[52]房俊芳,钟秀会.黄芩的药理研究概述[A].中药药理研究进展,2003,7:187～189
[53]房俊芳.中药黄芩、白术在胚胎着床、胚胎移植过程中的作用及机制研究:[硕士学位论文].保定:河北农业大学,2004
[54]梁培育.黄芪注射液体外对人精子运动参数的影响.中华男科学,2001,10(5):335～337
[55]邸冉,高建明.中药成分对小鼠 2-细胞胚胎体外生长发育的影响初探[J].北京农学院学报,2004,19(增):61～65
[56]傅文栋.小檗碱、黄芩苷、黄芪多糖对小鼠 2-细胞胚胎体外培养及移植效果的研究:[硕士学位论文].乌鲁木齐:新疆农业大学,2005
[57]徐平,川野佳代,刘丽均,等.实验大鼠、小鼠胚胎的缓慢冷冻和玻璃化冷冻的比较[J].上海交通大学学报(农业科学版),2002,20(4):261～265
[58]冯涛.动物胚胎冷冻保存技术的研究进展[J].中国草食动物,2004 专辑:58～60
[59]常万存,贾青,路兴中.以冷冻胚子和胚胎保存动物遗传资源的可行性[J].中国育牧杂志,1998, 34( 1) : 49～50
[60]包华琼,李跃民.哺乳动物胚胎冷冻的研究进展[J]. 草食家畜,2001, 4(113):1～5
[61]付永伦.人类卵母细胞冷冻保存[J].生殖与避孕,1996,16( 3):163～166
[62]李善国.人类胚胎和卵子的冷冻保存[J].生殖与避孕,1997,17(1):3～7
[63]Rall WF, Fahy GM.Ice-free cryopreservation of mouse embryos at -196℃ byvitrification[J].Nature,1985,313(6003):573～575
[64]张爱民编译.牛胚胎移植的现状及展望[J].草食家畜,1994,2:27～29
[65]王新压,窦忠英.影响哺乳动物胚胎冷冻效果的因素分析[J].黄牛杂志,1996,22( 2):40～42
[66]谭瑛,孙振权.家畜胚胎移植技术研究历史及应用现状[J].呼伦贝尔学院学报,2001,2(9):25～28
[67]黄莉,戴立军.小鼠胚胎移植初试[J].广州医学院学报,2001,4(29):36～38
[68]刘丽均,徐平.小鼠桑椹期、囊胚期胚胎移植方法的探讨[J].上海实验动物科学,2000,2(20):109～112
[69]Bolton VN, Wren ME, Parsons JH. Pregnancies after in vitro fertilization andtransfer of human blastocysts[J]. Fertil Steril,1991,55:830～832
[70]Marek D, Langley M, Gardner DK, et al. Introduction of blastocyst culture andtransfer for ail patients in vitro fertilization program[J]. Fertil Steril, 1999, 72:1035～1040
[71]Gardner DK, Phil D, Vella P, et al.Culture and transfer of human blastocystsimplantation rates and reduces the need for multiple embryo transfers[J]. Fertil Steril,1998, 69:84～88
[72]Toledo AA, Wright G, Jones AE, et al. Blatocyst transfer:a useful tool for reductionof high-order multiple gestations in a human assisted reproduction program[J]. Am J ObstetGynecol, 2000, 183: 377～379
[73]Barroso RP,Osuamkpe C,Nagamana M,et al.Nitric oxide inhibits developmentof embryos and implantation in mice[J].Mol Hum Reprod,1998,4(5):503～507
[74]Huang R,Zhu GJ. The Role Nitric Oxide in the Process of implantation inmice[J].Acta Univ Med Tongji,2001,30(1):42～45
[75]彭守静,陆仁康,俞丽华,等.菟丝子、仙茅、巴戟天对人精子体外运动和膜功能影响的研究[J].中国中西医结合杂志,1997,17(3):145
[76]曾金雄,戴西湖,刘建华,等.中药还精方对人精子运动及相关参数的影响[J].中国男科学杂志,2002,16(4):319
[77]姚石安.中药促排卵的研究[J].中国药业,1996,2:12
[78]刘玉鹏,刘梅,刘俊英,等.30 种中草药的抗氧化活性研究[J].烟台大学学报,2000,1(1):70～73
[79]先宏,吴可,孙存普.中药抗氧化活性的主要成分及其自由基清除作用[J].国外医学中医中药分册,2003,25(3):150～153
[80]阮娜,宋晓平. 补益药对体外培养细胞的影响.中兽医药研究进展(论文集) [C].陕西杨陵:西北农林科技大学出版社,2003,158～160
[81]邸冉,高建明.中药成分对小鼠 2-细胞胚胎体外生长发育的影响初探[J].北京农学院学报,2004,19(增):61～65
[82]傅文栋,高建明,陈武,等.小檗碱及黄芩苷对小鼠 2-细胞胚胎体外培养效果的初步研究[A].中国兽医医药杂志,2005(专辑):24:146～150
[83]傅文栋,孙玉成,索伦,等.小鼠发情周期观察与最佳超排时期的确定[J].北京农学院学报,2005,20(2):19～21
[84]Chatot CL,Lewis JL,Torris I,et al.One-minute exposure of 4-cell mouse embryosto glucose overcomes morula block in CZB medium[J].Mol Reprod Dev,1994,37(4):407～412
[85]庞战军,周玫,陈瑗.自由基医学研究方法[M].北京:人民生出版社,2000:224～226
[86]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[J].中国中西医结合杂志,2004,24(7):627～629
[87]杨桂云,王佩娟,贾晓斌,等.补肾活血汤对小鼠体外受精及其早期胚胎发育的影响[J].中国中西医结合杂志,2001,21(7):522～524
[88]陈骞,李尚为,唐海峰,等.瘦素对小鼠着床前胚胎发育影响的体外研究[J].四川大学学报(医学版),2004,35(6):806～808
[89]白照岱,刘凯,邴鲁军.表皮生长因子对小鼠早期胚胎体外发育影响的研究[J].山东大学学报(医学版),2004,42(2):150～158
[90]黄吴键,袁进,邓星,等.KSOM 培养基营养成分调整对小鼠植入前期胚体外发育的影响[J].第一军事医科大学学报[J],2005,25(3):256～261
[91]Berh B, Mooney S, Wen Y,et al. Preliminary experience with low concentration ofgranulocyte-macrophage colony-stimulating factor:a potential regulator in preimplantationmouse embryo development and apoptosis[J]. J Assist Reprod Genet,2005, 22(1):25～32
[92]Horng-Der TSAI,Chi-Chen Chang,Yao-Yuan HSIEH, et al.Effect of DifferentConcentrations of Recombinant Leukemia Inhibitory Factor on Different Development Stageof Mouse Embryo In Vitro[J].J Assist Reprod Genet,2000,17(6):352～355
[93]Manejwala FM,Kaji E,Sehultz RM.Development of activatable adenylate cyclasein the preimplantation mouse embryo and a role for camb in blastocoel formation[J].cell,1986,46(1):99～103
[94]Nonogaki T, NodaY, Goto Y, et al. Developmental block age of mouse embryoscaused by fatty acids[J].J Assist Reprod Genet,1994,11(9):482～488
[95]Shiimizu I, Ma Y R, Mizobuchi Y, et al.Effects of Sho-saiko-to, a Japanese herbalmedicine, on hepatic fibrosis in rats[J]. Hepatology, 1999, 29(1):149～160
[96]王砚宁,毕新岭,顾军,等.黄芩甙影响人成纤维细胞诱导型一氧化氮合酶蛋白表达研究[J].中国中西医结合皮肤性病学杂志,2003,2(3):152～154
[97]黎玉,万福生,李少华.川芎嗪对大鼠缺血心肌线粒体 NO 及自由基的影响[J].中医药学报,2004,32(2):47～49
[98]施月欢,邹秀兰,赵松滨.川芎嗪对大鼠缺血在灌注视网膜 SOD、MDA 和 NO水平及细胞凋亡的影响[J].眼科研究,2001,19(4):301～303
[99]晏雪生,彭亚琴,明安萍.川芎嗪注射液对体外培养软骨细胞影响的实验研究[J].中国中医骨伤科杂志,2002,10(1):15～17
[100]张兆辉,余绍祖,赵保路.四甲吡嗪对人类白细胞呼吸爆发氧代谢的影响[J].中国临床药理学与治疗学,2001,6(1):8～11
[101]Jiang JY, Geng DS,Tursonjan T,et al.Anti-inflammatory effect and mechanismof berberine[J].chin pharmacolBull,1998,14(5):434～437
[102]王志红,林菁.盐酸小檗碱对 HL-60 细胞增殖与分化的影响[J].中国药理学通报,2004,20(11):1305～1308
[103]杨青,薛立群,刘斌,等.维生素 E 对附植前小鼠胚胎发育与一氧化氮生成的影响[J].湖南农业大学学报(自然科学版),2003,29(4):288～290
[104]朱孝荣,彭长凌,袁红花.小鼠胚胎冷冻研究[J].徐州医学院学报,2005,25(1):45～47
[105]Sinclair KP,Young LE,Wilmut I,et al.In-utero overgrowth in ruminantsfollowing embryo culture:lessons from mice and a warning to men[J].Hum Reprod, 2000,15(Suppl 5):68～86
[106]Lane M . Mechanisms for managing cellular and homeostatic stress invitro[J].Theriogenology,2001,55:225～236
[107]Farin PW,Stockburger EM,Rodriguez KF,et al.Placental morphology is alteredfollowing transfer of bovine embryos produced invitro[J].Theriogenol,2000b,53:474～482
[108]Papadopoulos S,Rizos D,Duffy P,et al.Embryo survival and recipient pregnancyrates after transfer of fresh or vitrified ,in vivo or in vitro produced ovine blastocysts[J]. AnimReprod Sci,2002,74:35～44
[109]许建平,马庭元,闻良珍.川芎嗪治疗胎儿宫内生长迟缓中氧自由基与血栓素β2、6-酮-前列腺素 1α的相关性研究[J].中国中西医结合杂志,1998,18(5):265~268
[110]Cernakova M,Kostalova D.Antimicrobial activity of berberine a constituent ofMahonia aquifolium[J].Folia Microbiol,2002,47(4):375～378
[111]宋淑娟.黄连素治疗妊娠合并心律失常 16 例临床观察[J].吉林中医药,1997,5:19
[112]高雨农,高晓山.妊娠晚期服用盐酸小檗碱对新生儿血清胆红素影响的回顾性调查简报[J].中国中药杂志,2000,9(25):564～565
[113]Kevin S, Richter D, Dee C, et al.Quantitative grading of a human blastocyst:optimal inner cell mass size and shape[J].Fern Steri, 2001, 76:1157～1167
[114]孙艳香,姜国诚,雷俊英.小鼠囊胚玻璃化冷冻保存研究[J].广西师范大学学报(自然科学版),2002,20(3):73～75
[2]赵保路.氧自由基和天然抗氧化剂[M].北京:科学技术出版社,1999
[3]刘建国,王宪,陈明哲.一氧化氮和细胞因子之间的相互调节作用[J].生理科学进展,2000,31(1):61～64
[4]王雁.一氧化氮对女性生殖系统的影响[ J].首都医药,2003,10(4):44～45
[5]Rossellli M,Keller PJ,Dubey PK.Role of nitric oxide in biology,physiology andpathophysiology of reproduction[J].Hum Reprod Up date,1998,4(1):3～24
[6]郑荣梁,张红.自由基对人精子功能的调控[J].扬州大学学报(自然科学版),1999,35(3):135～138
[7]Dey SK,Komura F,Mukherjee A,et al.Cyclic AMP and Cyclic GMP in rabbitblastocysts[J].ReProd Fertil,1978,52(2):235～237
[8]王元兴,郎介金.动物繁殖学[M].南京:江苏科学技术出版社,1993,166～174
[9]Brief S,Chew BP.effect of vitamin A and β-carotene on reproductive performance ingilts[J].J Anim Sci,1985,60(4):998～1004
[10]Quinn P,Harlow GM.The effect of oxygen on the development of preimplantationmouse embryos in vitro[J].J Exp Zool,1978,206(1):73～80
[11]Flood MR,Wiebold JL.Glucose metabolsm by preimplantation pig embryos[J],JReprod Fert,1988,84(1):7～12
[12]Laloraya M,Kumar GP,Laloraya MM.A possible role of superoxide anion radicalin the process of blastocyst implantation in musculus.Biochem Biophys Res Commun,1989,161(2):762～770
[13]Nasr-Esfahani MH,Aitken JR,Johnson MH.hydrogen peroxide levels in mouseoocytes and early cleavage stage embryos developed in vitro or in vivo[J].Development,1990,109(2):501～507
[14]Noda Y,Watsumoto H,Umadka Y,et al.Involvement of superoxide radicals in themouse two-cell block[J].Mol Reprod Dev.1991,28(4):356～360
[15]Li JM,Foote RH,Simkin M.Development of rabbit zygotes cultured in protein-freemedium with catalase,taurine or superoxide dismutase[J].Biol Reprod,1993,49(1):33～37
[16]Payne SR,Munday R,Thompson JG.Addition of superoxide dismutase and catalasedoes not necessarily overcome developmental retardation of one-cell mouse embryo duringin-vitro culture[J].Reprod Fertil Dev,1992,4(2):167～174
[17]Thomas M,Jain S,Kumar GP,et al.A programmed oxyradical burst causes hatchingof mouse blastocysts[J].Journal of cell science,1997,110(14):1597～1602
[18]Kuo RC,Baxter GT,Thompson SH,et al.NO is necessary and sufficient for eggactivation at fertilization[J].Nature,2000,406(6796):633～636
[19]Novaro V,Gonzalez E,Rettori V,et al.Nitric oxide synthase regulation duringembryonic implantation[J].Reprod Fertil Dev,1997,9(5):557～564
[20]Duran-Reyes G,Gomez-Melendez MR,Brena G.Nitric oxide Synthase inhibitionsuppresses implantation and decreases cGMP concentration and protein peroxidation[J].LifeScience,1999,65(21):2259～2268
[21]Gouge RC,Marshburn P,Gordon BE,et al.Nitric oxide as a regulator embryonicdevelopment[J].Biol Reprod,1998,58(4):875～879
[22]Abe K,Matsuoka K,Inoue N,et al.Messenger RNA of neuronal nitric oxidesynthase is expressed and possibly functions in mouse oocytes and embryos duringpreimplantation development[J].Biomed Res,1999,20(1):61～65
[23]Athanassakis I,Aifantis I,Baritakis S,et al.Nitric Oxide production bypre-implantation embryo in response to embryotoxic factors.Cellular Physiology andBiochemistry,2000,10(3):169～176
[24]解启发,邓泽沛.NO 在小老鼠早期胚胎吸收过程中的作用[J].中国兽医学报,2002,22(1):90～93
[25]周思畅,周剑涛.一氧化氮介导细胞凋亡的分子基础[J].生命科学,2002,14(3):135～138
[26]Purcell TL,Given R,Chwalisz K,et al.Nitric oxide synthase distribution duringimplantation in the mouse[J].Mol Hum Reprod,1999,5(5):467～475
[27]Farina M,Ribeiro ML,Ogando D,et al.IL-1 alpha augments prostaglandinsynthesis in pregnant rat uteriby a nitric oxide mediated mechanism[J].Prostaglandins LeukotEssent Fatty Acids ,2000,62(4):243～247
[28]王建辰,章孝荣.动物生殖调控[M].安徽:安徽科学技术出版社.1998:102
[29]Biswas S,Kabir SN.Therole of Nitric oxide in the process of implantation inrats[J].Journal of Reproduction and Fertility,1998,114(1):157～161
[30]朱猛进,丁家酮.一氧化氮(NO)对动物妊娠生理的调节[J].贵州畜牧兽医,2000,24(1):10～11
[31]林毅宏.自由基与中药 BRM 效应[J].福建中医药,1992,(4):41
[32]Robak J . Flavonids are scavengers of super oxideanions Biochemical [J].Pharmacology,1988,37:837
[33]Husain SR.Hydroxyl radical scavenging activity of flavonoids[J].Phytochemistry,1987,26(9):2489
[34]吴文林,胡天喜.几种黄酮类化合物对羟自由基引起的损伤的保护作用.自由基生命科学进展(第 5 集)[M].北京:原子能出版社,1997,101
[35]康鑫,方允中,刘学英.茶多酚清除超氧化物自由基的效能及其对肿瘤细胞 DNA生物合成的影响.自由基生命科学进展(第 1 集)[M].北京:原子能出版社,1993,163
[36]宋洪涛,郭涛,宓鹤鸣.天然药物中的抗氧化剂[J].中草药,1991,22(7):331
[37]詹皓,沈竟民,姜桂荣.中药抗氧化作用的研究概况[J].中国中药杂志,1990,15(10):54
[38]姚文兵,高向东,周慧萍,等.大黄的生化学研究 XXXVI 波叶大黄多糖抗衰老作用的实验研究[J].中国药科大学学报,1991,22(3):167
[39]劳汉昌,张宝华,胡侦明.脊髓损伤早期三七总皂苷抗氧化作用的实验研究[J].云南医药,1995,16(5):355
[40]何凤慈,唐汝愚,姚丹帆.粉防已碱对急性炎症血管通透性和嗜中性白细胞功能的影响[J].中国药理学报,1989,10(3):249.
[41]王利津,徐强.黄连解毒汤的抗氧化作用研究[J].中国药科大学学报,2001,32(1):51～53
[42]龚跃新,孙云,林安平.补气法、补血法抗自由基操作的比较研究[J].中国中药杂志,1993,18(7):438
[43]张其兰,孙志福.复方何首乌抗氧化作用的研究[J].自由基生命科学进展(第 1集)[M].北京:原子能出版社,1993:189
[44]Guerin P,Mouatassim S,Menezo Y.Oxidative stress and protection against reactiveoxygen species in the preimplantation embryo and its surroundings[J].HumReprod Update,2001,7:175～189
[45]Noda Y,Goto Y,Umaoka Y,et al.Culture of Human Embryos in AlphaModification of Eagle's Mdium under Low Oxygen Tensionan Low Illumination[J].FertilSteril,1994,62:1022～1027
[46]Umaoka Y,Noda Y,Narimoto K,et al.Developmental Potentiality Of EmbryosCultured under Low Oxygen Tension with Superoxide Dismutase[J].J In Vitro Fert EmbryoTransf,1991,8(5):245～249
[47]Herrick JB.Injectable β-carotene Appears to Improve Reproduction[J].Feed stuff,1993,7:13～17
[48]吴师光,董孝国.一种古老的生物信息分子-退黑素[J].商丘师范学院学报,2003,19(2):102～103
[49]关钧.环核苷酸与中药效功能综述[J].浙江中医杂志,1989,244(5):234～236
[50]孙忠亲.中草药对环核苷酸影响的研究进展[J].中草药,1990,21(12):31～32
[51]阮娜,宋晓平.补益药对体外培养细胞的影响[A].中兽医药研究进展,2003,7:158～160
[52]房俊芳,钟秀会.黄芩的药理研究概述[A].中药药理研究进展,2003,7:187～189
[53]房俊芳.中药黄芩、白术在胚胎着床、胚胎移植过程中的作用及机制研究:[硕士学位论文].保定:河北农业大学,2004
[54]梁培育.黄芪注射液体外对人精子运动参数的影响.中华男科学,2001,10(5):335～337
[55]邸冉,高建明.中药成分对小鼠 2-细胞胚胎体外生长发育的影响初探[J].北京农学院学报,2004,19(增):61～65
[56]傅文栋.小檗碱、黄芩苷、黄芪多糖对小鼠 2-细胞胚胎体外培养及移植效果的研究:[硕士学位论文].乌鲁木齐:新疆农业大学,2005
[57]徐平,川野佳代,刘丽均,等.实验大鼠、小鼠胚胎的缓慢冷冻和玻璃化冷冻的比较[J].上海交通大学学报(农业科学版),2002,20(4):261～265
[58]冯涛.动物胚胎冷冻保存技术的研究进展[J].中国草食动物,2004 专辑:58～60
[59]常万存,贾青,路兴中.以冷冻胚子和胚胎保存动物遗传资源的可行性[J].中国育牧杂志,1998, 34( 1) : 49～50
[60]包华琼,李跃民.哺乳动物胚胎冷冻的研究进展[J]. 草食家畜,2001, 4(113):1～5
[61]付永伦.人类卵母细胞冷冻保存[J].生殖与避孕,1996,16( 3):163～166
[62]李善国.人类胚胎和卵子的冷冻保存[J].生殖与避孕,1997,17(1):3～7
[63]Rall WF, Fahy GM.Ice-free cryopreservation of mouse embryos at -196℃ byvitrification[J].Nature,1985,313(6003):573～575
[64]张爱民编译.牛胚胎移植的现状及展望[J].草食家畜,1994,2:27～29
[65]王新压,窦忠英.影响哺乳动物胚胎冷冻效果的因素分析[J].黄牛杂志,1996,22( 2):40～42
[66]谭瑛,孙振权.家畜胚胎移植技术研究历史及应用现状[J].呼伦贝尔学院学报,2001,2(9):25～28
[67]黄莉,戴立军.小鼠胚胎移植初试[J].广州医学院学报,2001,4(29):36～38
[68]刘丽均,徐平.小鼠桑椹期、囊胚期胚胎移植方法的探讨[J].上海实验动物科学,2000,2(20):109～112
[69]Bolton VN, Wren ME, Parsons JH. Pregnancies after in vitro fertilization andtransfer of human blastocysts[J]. Fertil Steril,1991,55:830～832
[70]Marek D, Langley M, Gardner DK, et al. Introduction of blastocyst culture andtransfer for ail patients in vitro fertilization program[J]. Fertil Steril, 1999, 72:1035～1040
[71]Gardner DK, Phil D, Vella P, et al.Culture and transfer of human blastocystsimplantation rates and reduces the need for multiple embryo transfers[J]. Fertil Steril,1998, 69:84～88
[72]Toledo AA, Wright G, Jones AE, et al. Blatocyst transfer:a useful tool for reductionof high-order multiple gestations in a human assisted reproduction program[J]. Am J ObstetGynecol, 2000, 183: 377～379
[73]Barroso RP,Osuamkpe C,Nagamana M,et al.Nitric oxide inhibits developmentof embryos and implantation in mice[J].Mol Hum Reprod,1998,4(5):503～507
[74]Huang R,Zhu GJ. The Role Nitric Oxide in the Process of implantation inmice[J].Acta Univ Med Tongji,2001,30(1):42～45
[75]彭守静,陆仁康,俞丽华,等.菟丝子、仙茅、巴戟天对人精子体外运动和膜功能影响的研究[J].中国中西医结合杂志,1997,17(3):145
[76]曾金雄,戴西湖,刘建华,等.中药还精方对人精子运动及相关参数的影响[J].中国男科学杂志,2002,16(4):319
[77]姚石安.中药促排卵的研究[J].中国药业,1996,2:12
[78]刘玉鹏,刘梅,刘俊英,等.30 种中草药的抗氧化活性研究[J].烟台大学学报,2000,1(1):70～73
[79]先宏,吴可,孙存普.中药抗氧化活性的主要成分及其自由基清除作用[J].国外医学中医中药分册,2003,25(3):150～153
[80]阮娜,宋晓平. 补益药对体外培养细胞的影响.中兽医药研究进展(论文集) [C].陕西杨陵:西北农林科技大学出版社,2003,158～160
[81]邸冉,高建明.中药成分对小鼠 2-细胞胚胎体外生长发育的影响初探[J].北京农学院学报,2004,19(增):61～65
[82]傅文栋,高建明,陈武,等.小檗碱及黄芩苷对小鼠 2-细胞胚胎体外培养效果的初步研究[A].中国兽医医药杂志,2005(专辑):24:146～150
[83]傅文栋,孙玉成,索伦,等.小鼠发情周期观察与最佳超排时期的确定[J].北京农学院学报,2005,20(2):19～21
[84]Chatot CL,Lewis JL,Torris I,et al.One-minute exposure of 4-cell mouse embryosto glucose overcomes morula block in CZB medium[J].Mol Reprod Dev,1994,37(4):407～412
[85]庞战军,周玫,陈瑗.自由基医学研究方法[M].北京:人民生出版社,2000:224～226
[86]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[J].中国中西医结合杂志,2004,24(7):627～629
[87]杨桂云,王佩娟,贾晓斌,等.补肾活血汤对小鼠体外受精及其早期胚胎发育的影响[J].中国中西医结合杂志,2001,21(7):522～524
[88]陈骞,李尚为,唐海峰,等.瘦素对小鼠着床前胚胎发育影响的体外研究[J].四川大学学报(医学版),2004,35(6):806～808
[89]白照岱,刘凯,邴鲁军.表皮生长因子对小鼠早期胚胎体外发育影响的研究[J].山东大学学报(医学版),2004,42(2):150～158
[90]黄吴键,袁进,邓星,等.KSOM 培养基营养成分调整对小鼠植入前期胚体外发育的影响[J].第一军事医科大学学报[J],2005,25(3):256～261
[91]Berh B, Mooney S, Wen Y,et al. Preliminary experience with low concentration ofgranulocyte-macrophage colony-stimulating factor:a potential regulator in preimplantationmouse embryo development and apoptosis[J]. J Assist Reprod Genet,2005, 22(1):25～32
[92]Horng-Der TSAI,Chi-Chen Chang,Yao-Yuan HSIEH, et al.Effect of DifferentConcentrations of Recombinant Leukemia Inhibitory Factor on Different Development Stageof Mouse Embryo In Vitro[J].J Assist Reprod Genet,2000,17(6):352～355
[93]Manejwala FM,Kaji E,Sehultz RM.Development of activatable adenylate cyclasein the preimplantation mouse embryo and a role for camb in blastocoel formation[J].cell,1986,46(1):99～103
[94]Nonogaki T, NodaY, Goto Y, et al. Developmental block age of mouse embryoscaused by fatty acids[J].J Assist Reprod Genet,1994,11(9):482～488
[95]Shiimizu I, Ma Y R, Mizobuchi Y, et al.Effects of Sho-saiko-to, a Japanese herbalmedicine, on hepatic fibrosis in rats[J]. Hepatology, 1999, 29(1):149～160
[96]王砚宁,毕新岭,顾军,等.黄芩甙影响人成纤维细胞诱导型一氧化氮合酶蛋白表达研究[J].中国中西医结合皮肤性病学杂志,2003,2(3):152～154
[97]黎玉,万福生,李少华.川芎嗪对大鼠缺血心肌线粒体 NO 及自由基的影响[J].中医药学报,2004,32(2):47～49
[98]施月欢,邹秀兰,赵松滨.川芎嗪对大鼠缺血在灌注视网膜 SOD、MDA 和 NO水平及细胞凋亡的影响[J].眼科研究,2001,19(4):301～303
[99]晏雪生,彭亚琴,明安萍.川芎嗪注射液对体外培养软骨细胞影响的实验研究[J].中国中医骨伤科杂志,2002,10(1):15～17
[100]张兆辉,余绍祖,赵保路.四甲吡嗪对人类白细胞呼吸爆发氧代谢的影响[J].中国临床药理学与治疗学,2001,6(1):8～11
[101]Jiang JY, Geng DS,Tursonjan T,et al.Anti-inflammatory effect and mechanismof berberine[J].chin pharmacolBull,1998,14(5):434～437
[102]王志红,林菁.盐酸小檗碱对 HL-60 细胞增殖与分化的影响[J].中国药理学通报,2004,20(11):1305～1308
[103]杨青,薛立群,刘斌,等.维生素 E 对附植前小鼠胚胎发育与一氧化氮生成的影响[J].湖南农业大学学报(自然科学版),2003,29(4):288～290
[104]朱孝荣,彭长凌,袁红花.小鼠胚胎冷冻研究[J].徐州医学院学报,2005,25(1):45～47
[105]Sinclair KP,Young LE,Wilmut I,et al.In-utero overgrowth in ruminantsfollowing embryo culture:lessons from mice and a warning to men[J].Hum Reprod, 2000,15(Suppl 5):68～86
[106]Lane M . Mechanisms for managing cellular and homeostatic stress invitro[J].Theriogenology,2001,55:225～236
[107]Farin PW,Stockburger EM,Rodriguez KF,et al.Placental morphology is alteredfollowing transfer of bovine embryos produced invitro[J].Theriogenol,2000b,53:474～482
[108]Papadopoulos S,Rizos D,Duffy P,et al.Embryo survival and recipient pregnancyrates after transfer of fresh or vitrified ,in vivo or in vitro produced ovine blastocysts[J]. AnimReprod Sci,2002,74:35～44
[109]许建平,马庭元,闻良珍.川芎嗪治疗胎儿宫内生长迟缓中氧自由基与血栓素β2、6-酮-前列腺素 1α的相关性研究[J].中国中西医结合杂志,1998,18(5):265~268
[110]Cernakova M,Kostalova D.Antimicrobial activity of berberine a constituent ofMahonia aquifolium[J].Folia Microbiol,2002,47(4):375～378
[111]宋淑娟.黄连素治疗妊娠合并心律失常 16 例临床观察[J].吉林中医药,1997,5:19
[112]高雨农,高晓山.妊娠晚期服用盐酸小檗碱对新生儿血清胆红素影响的回顾性调查简报[J].中国中药杂志,2000,9(25):564～565
[113]Kevin S, Richter D, Dee C, et al.Quantitative grading of a human blastocyst:optimal inner cell mass size and shape[J].Fern Steri, 2001, 76:1157～1167
[114]孙艳香,姜国诚,雷俊英.小鼠囊胚玻璃化冷冻保存研究[J].广西师范大学学报(自然科学版),2002,20(3):73～75