卵母细胞玻璃化冷冻对胚胎发育及妊娠结局的影响

目的探讨玻璃化冷冻对卵母细胞体外受精、胚胎发育以及妊娠结局的影响。方法将193枚玻璃化冷冻卵母细胞解冻复苏,采用卵胞浆内单精子注射法受精,观察胚胎发育形态,经72h体外培养后移植。结果成熟卵母细胞解冻复苏率为75．6％,正常受精率为72．6％,卵裂率为88．7％,优质胚胎率为48．9％。共移植19例,9例妊娠,已分娩2例,临床妊娠率为47．3％。新鲜周期与冻卵解冻周期相比,卵裂率、可移植胚胎率和优质胚胎率均无显著性差异（P〉0．05）。结论玻璃化冷冻方法对成熟卵母细胞的胚胎发育及妊娠结局无显著影响。     

35岁以下患者长方案促排卵获卵数对体外受精-胚胎移植结局的影响

目的探讨体外受精-胚胎移植（IVF-ET）周期最佳获卵数及其对妊娠结局的影响。方法回顾性分析我院2009年9月至2010年8月35岁以下行长方案IVF-ET治疗的544个获卵周期的资料。比较不同获卵数的受精率、卵裂率、优胚率、临床妊娠率及并发症等指标的差异。结果不同获卵数各组间促性腺激素（Gn）刺激天数、Gn用量、卵裂率、优胚率无明显差别（P〉0．05）,获卵数〉25个组受精率低于其他各组（P〈0．05）。随获卵数的增加,冷冻胚胎数、注射人绒毛膜促性腺激素（hCG）日血雌二醇（E2）值及卵巢过度刺激综合征（OHSS）发生率增加,差异有统计学意义（P〈0．05）。临床妊娠率随获卵数增加逐渐增加,当获卵数〉15个,临床妊娠率较前下降,累积妊娠率随获卵数增加而增加,当获卵数〉25个,临床妊娠率和累积妊娠率明显下降。结论获得适量的卵有利于获得较满意的妊娠结局。     

卵子玻璃化冷冻在体外受精-胚胎移植中的初步应用

目的:探讨体外受精-胚胎移植(IVF-ET)治疗中的卵子冷冻保存临床应用的可行性及其指征和方法,以提高卵子利用率和减少卵子的废弃。方法:在我院生殖医学中心进行IVF-ET治疗的患者,在取卵日因男方因素无法进行受精者或获卵数超过25枚,进行知情同意后对部分卵子进行玻璃化冷冻保存。将冷冻保存卵子解冻后继续使用,或捐赠给需要受卵患者。结果:2009年10月至2011年6月,已有7例患者的卵子进行了复苏。共复苏卵子53枚,存活44枚,复苏率83.02%;有41枚MⅡ卵子行ICSI,受精32枚,受精率72.73%;形成30枚胚胎,卵裂率93.75%,获有效胚胎23枚。9个移植周期移植胚胎16枚,临床妊娠2例,分娩3个健康婴儿,胚胎种植率18.75%,活产率22.22%。剩余冷冻胚胎7枚。结论:卵子的玻璃化冷冻保存,可以获得比较满意的复苏率和受精率;将获卵较多的患者进行部分卵子的冷冻保存可以提高卵子的利用率,可以作为临床治疗的一个可选择的方法。     

褪黑素对老化卵母细胞体外发育能力的改善效果评价

目的探讨体外培养阶段(IVC)添加褪黑素(MT)对卵母细胞老化引起发育受损的改善效果。方法采用不同浓度的过氧化氢(H_2O_2)处理小鼠MII期卵母细胞诱导老化,浓度分别为0(对照组)、10、50、100、150μmol/L,体外受精(IVF)后统计各组二细胞率、囊胚形成率,检测老化卵母细胞的线粒体活性及丰度(Mitotracker Red、JC-1)、活性氧(ROS)水平、线粒体拷贝数等指标;在100μmol/L H_2O_2处理条件下,老化卵母细胞在IVF后的培养阶段分别添加不同浓度褪黑素(10-5、10-7、10-9 mol/L),统计二细胞率、囊胚率,并且分别检测各组获得囊胚的细胞数及凋亡率。结果不同浓度H_2O_2诱导卵母细胞老化后,囊胚发育率随H_2O_2浓度的升高而降低,50μmol/L和100μmol/L H_2O_2组囊胚形成率分别为(26.27±0.06)%和(28.46±3.45)%,相比对照组的(34.90±1.77)%显著降低,差异有统计学意义(P0.05);在H_2O_2诱导老化卵母细胞中,100μmol/L、150μmol/L H_2O_2浓度时,ROS水平相比对照组显著升高,差异具有统计学意义(P0.05);而活性线粒体的丰度及拷贝数呈现下降趋势,膜电位呈上升趋势,但相比对照组无显著性差异(P0.05);100μmol/L H_2O_2处理诱导卵母细胞老化后,在培养液中添加10-9 mol/L褪黑素组与老化对照组相比,囊胚率[(29.42±2.39)%vs.(20.87±4.12)%]、囊胚细胞数(39.36±9.78vs.37.91±4.25)均显著升高,凋亡率显著下降(2.57%vs.3.18%),差异均有统计学意义(P0.05);并且这些指标均达到与未经老化处理组的相似发育水平。结论老化卵母细胞体外受精后发育率和发育质量偏低的现象,可以通过在受精后体外培养阶段添加褪黑素得到改善。     

Effects of paternal cigarette smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation in rats

We evaluated the effects of paternal smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation. Rats of group A were exposed to cigarette smoke for 10 weeks. Rats of group B were exposed to the smoke of incense sticks for 10 weeks. Rats of group C served as a control group. Rats of group D were exposed to cigarette smoke for 7 weeks only. Experimental period was 10 weeks in all groups. At the end of the experimental period serum testosterone responses to human chorionic gonadotropin stimulation, androgen-binding protein activity in testicular cytosols, epididymal sperm motility, and oocyte fertilization rate, oocyte cleavage rate, and blastocyst development rate after in vitro fertilization (IVF) trials were significantly smaller in group A compared with groups B and C. In contrast, fertilization rate, cleavage rate, and blastocyst development rate after intracytoplasmic sperm injection (ICSI) procedures were not significantly different among groups A, B, C, and D. Both after IVF trials and ICSI techniques, the proportion of the alive offspring to the number of transferred oocytes was significantly smaller in group A than in groups B and C. Cigarette smoke-exposure results in a secretory deficiency of Leydig and Sertoli cells leading to an impaired epididymal sperm maturation process and diminished capacity of spermatozoa to penetrate oocytes. In addition paternal cigarette smoke exposure affects the embryonic ability for implantation.     

人体外与体内成熟卵母细胞发育潜能及妊娠结局的观察

目的观察体外成熟(IVM)与体内成熟人卵母细胞的发育潜能和妊娠结局。方法69例多囊卵巢或多囊卵巢综合征不育患者,分别行IVM治疗49个周期(IVM组)和体外受精-胚胎移植(IVF-ET)治疗29个周期(IVF组)。结果IVM组卵母细胞成熟率为65.4%。IVM组和IVF组受精率和卵裂率分别为62.1%和77.2%,60.2%和91.2%,两组受精率无显著性差异(P>0·05),IVM组较IVF组卵裂率降低非常显著(P<0.01)。IVM组和IVF组临床妊娠率和种植率分别为26.5%和12.6%,41.4%和22.9%,IVM组临床妊娠率和种植率较低,但两组无显著性差异(P>0.05)。IVM组和IVF组卵母细胞受精后D2≥4细胞和D3≥6细胞胚胎数占受精卵数(2PN)的比例分别为24.5%和19.4%,67.0%和66.4%,IVM组较IVF组胚胎发育速度慢,两者有非常显著性差异(P<0.01)。IVM组D3优质胚胎形成率(23.7%)较IVF组(58.2%)为低,亦有非常显著性差异(P<0.01)。结论人卵母细胞IVM将成为一种选择性的辅助生殖技术。人卵IVM较体内成熟卵母细胞受精后2PN至卵裂期发育阻滞发生率高,胚胎发育速度慢和优质胚胎形成率低。     

Intracytoplasmic sperm injection in cases with history of in vitro fertilization failure

Aim: To evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure. Methods: Two groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase Ⅱ oocytes: 0~30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35~37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase Ⅱ oocytes were selected for microinjection. Results: Fertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 %±21.1 % vs. 73.9 %±19.2 %), cleavage rate (85.1 %±19.3 % vs. 82.7 %±22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups. Conclusion: ICSI is a valuable method for     

卵子激活剂对无精子症患者冻融睾丸精子ICSI后妊娠结局的影响

目的研究卵子激活剂CultActive在临床中能否提高无精子症患者冻融睾丸精子行卵胞浆内单精子显微注射(ICSI)后的妊娠结局。方法选择2015年1月~2019年12月我院收治的188例行冻融睾丸精子ICSI助孕治疗的无精子症患者作为研究对象,按照患者ICSI后是否进行应用卵子激活剂CultActive随机分为冻融睾丸精子组(n=107)和卵子激活组(n=81),对照组为新鲜睾丸精子组(n=129),分别比较了三组患者在年龄、不孕年限、促性腺激素用量、促性腺激素使用天数等临床资料的差异以及获卵数、受精率、2PN率、分裂率、优胚率、可利用胚胎率、临床妊娠率等胚胎发育情况的差异。结果三组患者在年龄、不孕年限、促性腺激素用量、促性腺激素使用天数、获卵数上均无统计学差异(P>0.05)。B组在2PN率、优胚率、可利用胚胎率上均高于A、C组,但无统计学差异(P>0.05)。三组患者在分裂率上也无统计学差异(P>0.05)。B组在受精率上明显高于C组(P<0.05),且显著高于A组(p<0.01)。B组在临床妊娠率上显著高于A、C组(p<0.01)。结论卵子激活剂CultActive可以提高冻融睾丸精子ICSI效率,值得在临床中推广。     

两步极体活检对小鼠卵母细胞体外受精和胚胎发育的影响

目的评价两步活检一、二极体对小鼠卵母细胞体外受精(IVF)和相应胚胎发育的影响。方法8～12周龄雌、雄昆明小鼠。雌鼠促排卵,取成熟卵母细胞行卵胞浆内单精子注射(ICSI),序贯培养胚胎至囊胚孵出。分别于ICSI后激光透明带打孔活检第一极体,两步活检一、二极体,在受精后打孔同时活检一、二极体。比较各处理组与对照组受精、卵裂、优质胚胎、囊胚形成、扩张和孵出情况。结果两步活检一、二极体和同时活检一、二极体受精率、卵裂率、优质胚胎率和囊胚形成率与对照组均无明显差异。启动孵出率明显高于对照组;完全孵出率虽高于对照组,但无统计学意义。结论一、二极体活检不影响小鼠卵母细胞IVF和相应胚胎发育,而且可能有助于胚胎孵出,是安全可行的植入前遗传学诊断(PGD)活检取材方法,以两步活检法为最佳极体活检方案。     

显微授精穿刺方式与胚胎受精率及闭锁率相关性研究

目的探讨显微授精时由于卵膜弹性不同产生的不同破膜方式对卵母细胞受精率及卵母细胞闭锁率的影响。方法回顾性分析在本中心行显微授精治疗的患者共1589个卵母细胞，根据显微注射针穿刺卵膜的方式分为两组：一组1250例为卵膜弹性正常组，另一组339例为卵膜无弹性组，比较不同分组间的受精率差异，及穿刺后卵母细胞闭锁率。结果根据卵膜弹性分成的两组受精率无差异，卵膜有弹性组受精率为88．2％，卵膜无弹性组受精率87、3％，P〉0.05；穿刺后卵细胞闭锁的比率在卵膜无弹性组为17.1％，而在卵膜有弹性组的此比率为5.4％，两者有明显的统计学差异，P〈0.001。结论显微授精的受精率与穿刺时卵子的质量密切相关，所以选择恰当的取卵时机获取成熟度适当的卵子是提高显微授精受精率的关键。     

Human sperm movement assessed with the Hamilton-Thorn motility analyzer and in vitro fertilization

Summary. The relationship between sperm movement characteristics obtained by computerized analysis and the in vitro fertilization rates of human oocytes was studied. In 144 consecutive in vitro fertilization treatments a sample of prepared semen was analysed by a Hamilton-Thorn Motility Analyzer. In addition a visual estimation of sperm count and motility was made. Significant correlations with the fertilization rate were found for all visual parameters. Of the computerized measurements, the mean velocities of motile spermatozoa and the concentration of motile cells were significantly correlated. The average path velocity correlated best ( r = 0.42, P < 0.001). There was no relationship between the percentage of motile sperm showing hyperactivated movement and the fertilization rate. A forward stepwise logistic regression analysis selected the following variables of predictive value for fertilization: average path velocity, male factor infertility as indication for in vitro fertilization, motility and concentration, as measured by the Hamilton-Thorn analyzer. A logistic regression model to predict the cases with low (< 0.2) or high fertilization rates, included the average path velocity as a significant variable and classified the samples with 90% overall accuracy. In conclusion: movement characteristics of spermatozoa in culture medium, especially the average path velocity are of prognostic value in prediction of human oocyte fertilization rates.     

Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection （ICSI） using ejaculated and testicular spermatozoa

AIM: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). METHODS: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. RESULTS: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. CONCLUSION: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.     

常规体外受精失败卵母细胞行早期补救卵胞浆内显微注射的临床应用价值

目的探讨常规体外受精-胚胎移植（IVF—ET）中受精障碍患者行早期补救卵胞浆内单精子注射（ICSI）的可行性。方法回顾性分析我院生殖中心2007年1月至2009年7月常规IVF—ET治疗877周期,其中2008年3月至2009年7月期间开展短时受精并对受精障碍周期行早期补救ICSI的546周期作为研究组,2007年1月至2008年2月期间未开展短时受精、早期补救ICSI的331周期作为对照组。研究组通过IVF后6h观察卵母细胞是否排出第2极体评估受精,对完全未受精和低受精周期中未见第二极体排出的成熟卵母细胞立即行早期补救ICSI,比较两组临床及实验室指标。研究组中行早期补救ICSI的70周期实验室指标和临床指标与同期179个常规ICSI周期相比较。结果研究组通过早期补救ICSI,受精率、周期冷冻率、优质胚胎率均比对照组显著提高（P〈O．01）,因受精失败取消移植率显著降低（P〈O．01）。早期补救ICSI周期受精率、可移植胚胎率、临床妊娠率及胚胎种植率与常规ICSI相似,正常受精率、卵裂率和优质胚胎率显著低于常规ICSI,≥3原核（PN）异常受精的比率较常规ICSI略有升高,但无显著性差异。结论IVF后6h行早期补救ICSI能提高常规IVF卵的利用率,并获得与常规ICSI相似的临床妊娠结局。     

Minimal requirements for ejaculates used for in vitro fertilization

Minimal requirements for ejaculates to be used for in vitro fertilization of human oocytes are defined. Experimental results obtained a total concentration of at least 5 X 10(6) spermatozoa per ml are required, with at least 30% of the sperm showing normal motility and normal morphology in the differential spermiocytogram. The total volume of ejaculate should not be less than 1 ml. A total of 140 ejaculates from patients taking part in the in vitro fertilization program were examined. In 60 ejaculates (42.9%) bacteria concentrations of greater than 10(5)/ml were found. Eighty (57.1%) of the probes were negative or had bacteria concentrations less than 10(4)/ml. Mycoplasma was found in 30 of the examined ejaculates (21.4%) and the second most frequent group of microorganisms were enterococci found in 15 patients (10.7%). Anaerobic organisms in concentrations of more than 10(5)/ml were not detected. After insemination of one or more pelviscopically obtained oocytes with sperm of the husband in 14 out of 22 cases (63.6%) with bacteria concentrations greater than 10(5)/ml the cleavage of at least one oocyte occurred leading to an embryo-transfer rate of 63.6%. In 14 of 31 ejaculates with negative bacteriological findings (45.2%) at least one of the inseminated oocytes was fertilized. Direct effect of microorganism contaminated sperm on the human in vitro fertilization system resulting in reduced fertilization rates cannot be demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Oocyte membrane maturation and oocyte-sperm relationship: transmission electron microscopy study

The success of in vitro maturation (IVM) depends greatly on the acquisition of immature oocytes. Immature oocytes in prophase I (PI) and metaphase I (MI), aspirated after controlled ovarian hyperstimulation, were incapable of fertilization, leading to a lower fertilization rate. Therefore, they must be evaluated on a fine structure level for their in vitro maturation (IVM) processes and their relationship with sperm. Oocyte membrane maturation and oocyte-sperm relationship were studied using transmission electron microscopy. A total of 55 human oocytes obtained from 20 patients at various times and 83 oocytes obtained from the dissected ovarians of female Wistar rats were used for transmission electron microscopy (TEM) evaluation. Despite being in either prophase I and metaphase I or in metaphase II, the oocytes were not fertilized after 48 h of incubation. At the various stages of maturation between PI and MII, the number and the size of microvilluses on the oocyte membrane increased as MII approached and decreased after full maturation. Oocyte activation was related to oocyte membrane maturation and has an effect on the oocyte sperm penetration.     

常规体外受精失败周期早补救卵胞浆内单精子注射与部分-卵胞浆内单精子注射受精方式的回顾性分析

目的比较早补救卵胞浆内单精子注射（ICSI）与部分（Half）-ICSI两种授精方式在体外受精（IVF）助孕中的应用。方法回顾性分析76个常规IVF发生受精失败的移植周期。其中30个周期为早补救ICSI授精方式（早补救ICSI组）,46个周期为Half-ICSI授精方式（Half-ICSI组）。分析受精情况、胚胎质量、种植率、妊娠率、抱婴回家率、出生缺陷和卵母细胞利用情况。结果早补救ICSI组与Half—ICSI组的双原核（2PN）受精率、多PN受精率、1PN受精率、优质胚胎率、胚胎种植率、妊娠率、抱婴回家率、出生缺陷无显著差异（P〉0．05）。但早补救ICSI组卵母细胞利用率高于Half-ICSI组,平均受精卵数分别为（6．30±2．96）和（4．20±1．75）（P〈0．001）,胚胎数分别为（6．20±2．89）和（4．11±1．68）（P〈0．001）,冷冻胚胎数分别为（3．20±2．76）和（1．56±1．68）（P〈0．01）、冷冻周期率分别为76．67％和50．00％（P〈0．05）。结论在常规IVF完全受精失败的周期,早补救ICSI授精方式与Half-ICSI授精方式的受精率、胚胎质量和妊娠结局类似,但前者有更好的卵母细胞利用率和更多可利用胚胎。     

卵母细胞发育潜能与颗粒细胞凋亡关系的初步研究

目的　探讨卵母细胞发育潜能与颗粒细胞凋亡之间的关系。　方法　将卵母细胞在体外培养 28~30 h后,取其颗粒细胞,采用TUNEL法进行凋亡率的分析,并与卵子的成熟情况和卵子的评分及形成胚胎结果进行对照。　结果　经培养后,随着卵母细胞成熟度的降低,凋亡率升高;而且随着卵母细胞评分和发育潜能的降低,颗粒细胞凋亡率逐渐升高。　结论　卵子发育潜能与颗粒细胞凋亡密切相关,通过测定颗粒细胞的凋亡率可预测未成熟卵母细胞的发育潜能。     

The effect of nitrous oxide on in vitro fertilization success rate

The authors studied the effect of nitrous oxide on success rates for in vitro fertilization and pregnancy in women undergoing laparoscopy for oocyte retrieval. Ninety-eight patients in an in vitro fertilization program were randomly assigned to an anesthetic regimen including either 0.7% (end-tidal) isoflurane with 60% nitrous oxide in oxygen, or 1.4% (end-tidal) isoflurane in oxygen. Success rates for fertilization and pregnancy in 44 additional patients who declined randomization were also studied. Among the 51 randomized patients who did not receive nitrous oxide, 192 oocytes were obtained and 122 fertilized (63.5%), resulting in eight pregnancies (16.3%). From the 47 randomized patients given nitrous oxide, 168 oocytes were retrieved and 114 fertilized (67.9%), resulting in nine pregnancies (19.1%). No significant differences between rates of fertilization or pregnancy emerged between groups. Such differences would have been found with an 80% probability had nitrous oxide had a 20% effect on oocyte fertilization.     

ICSI treatment of severe male infertility can achieve prospective embryo quality compared with IVF of fertile donor sperm on sibling oocytes

Azoospermia, cryptozoospermia and necrospermia can markedly decrease the ability of males to achieve pregnancy in fertile females. However, patients with these severe conditions still have the option to be treated by intracytoplasmic sperm injection (ICSI) to become biological fathers. This study analyzed the fertilization ability and the developmental viabilities of the derived embryos after ICSI treatment of the sperm from these patients compared with in vitro fertilization (IVF) treatment of the proven-fertile donor sperm on sibling oocytes as a control. On the day of oocyte retrieval, the number of sperm suitable for ICSI collected from two ejaculates or testicular sperm extraction was lower than the oocytes, and therefore, excess sibling oocytes were treated by IVF with donor sperm. From 72 couples (73 cycles), 1117 metaphase II oocytes were divided into 512 for ICSI and 605 for IVF. Compared with the control, husbands’ sperm produced a lower fertilization rate in nonobstructive azoospermia (65.4% vs 83.2%; P < 0.001), crytozoospermia (68.8% vs 75.5%; P < 0.05) and necrospermia (65.0% vs 85.2%; P < 0.05). The zygotes derived in nonobstructive azoospermia had a lower cleavage rate (96.4% vs 99.4%; P < 0.05), but the rate of resultant good-quality embryos was not different. Analysis of the rates of cleaved and good-quality embryos in crytozoospermia and necrospermia did not exhibit a significant difference from the control. In conclusion, although the sperm from severe male infertility reduced the fertilization ability, the derived embryos had potential developmental viabilities that might be predictive for the expected clinical outcomes.     

罗比卡因和利多卡因对小鼠体外受精和早期胚胎发育的影响

目的 :应用昆明小鼠胚胎体外培养模式和小鼠体外受精模型探讨罗比卡因与利多卡因对体外受精和早期胚胎发育的影响。方法 :( 1 ) 2 -细胞期胚胎培养 :取 4～ 6周昆明雌鼠 ,下午 4～ 6时腹腔注射孕马血清 ( PMSG) 5IU,48h后腹腔注射人绒毛膜促性腺激素 ( h CG) 5IU超排卵 ,随即雌雄合笼。次日早上 9时检查阴栓 ,阳性者于注射 h CG42h后取 2 -细胞期胚胎 ,分别放入含 1、1 0及 1 0 0μg/ ml不同浓度的罗比卡因和利多卡因的M1 6培养液中培养 ,培养液中加入输卵管上皮 ,72 h后观察桑椹胚或囊胚形成率。 ( 2 )体外受精 :从 F1代 ( C57× CBA)杂交雄鼠附睾和输精管取精子 ,从昆明雌鼠取卵母细胞 ,卵母细胞体外受精前暴露于不同浓度的利多卡因和罗比卡因 3 0 min,统计体外受精率。结果 :罗比卡因只在高浓度 1 0 0 μg/ ml时对 2 -细胞至囊胚期的胚胎发育产生明显抑制影响。利多卡因则在 1μg/ ml的浓度即对 2 -细胞期胚胎发育产生明显抑制。而 1 0和 1 0 0μg/ ml的利多卡因和罗比卡因均未对体外受精率有明显影响。结论 :提示罗比卡因可能用于体外受精技术中有关配子和合子取出或植入时的麻醉。