外科取精术后行卵细胞胞浆内单精子注射治疗梗阻性无精子症

Palermo等于1992年首次报道卵细胞胞浆内单精子注射(ICSI)获得妊娠成功,为男性不育症治疗提供了新的方法[1]。同时,先进的外科附睾或睾丸取精术(microsurgical epididymal sperm aspiration,MESA/testicular sperm extraction,TESE)彻底改变了无精子症治疗的乏术局面。采用该方法只要在男性生殖道或睾丸内发现并分离到精子,利用ICSI技术就能获得高度有效的妊娠率。我们于2001年12月     

Effect of traditional Chinese herbs combined with low dose human menopausal gonadotropin applied in frozen thawed embryo transfer

Objective： To assess embryo implantation rate （IR） and pregnancy rate （PR） in women who received Bushen Wengong Decoction （补肾温宫汤, BSWGD）, a Chinese herbal formula, combined with low dose of human menopausal gonadotropin （hMG） prior to frozen-thawed embryo transfer （FET）. Methods： A total of 262 subjects （674 transferred embryos） who received FET were analyzed retrospectively. In them, 122 women were under 30 years old, 106 between 30-35 years and 32 over 35 years. The 85 subjects with normal ovulation were assigned to Group A, the natural menstruation cycling group, on whom no pre-transfer treatment was applied. The other 177 subjects with abnormal ovulation were assigned to Group B, and subdivided, according to the pre-transfer treatment they received, into three groups, Group B1 （50 cases） received BSWGD, Group B2 （58 cases） received hMG and Group B3 （69 cases） received both BSWGD and low dose hMG. The IR and PR of FET in the four groups were compared time on PR of FET were compared also. Besides, the influencing factors and the effect of the embryo cryoto FET were analyzed. Results： IR and PR were significantly higher in all age sects of Group B3 than those in Group A, showing significant difference （ P〈0.05）. IR and PR in subjects in age sects of 〈30 years and 〉 35 years in group B3 were significantly higher than those in Group B1 （ P〈0.05）, but no significant difference was shown in the two parameters between Group B 2 and Group B3 （P〉0.05）. PR in the subjects who received embryos with cryo-time of 〉 200 days was significantly lower than that in those with cryo-time of ~ 100 days （P〈0.05）. Embryo cryo-time, endometrial thickness, use of BSWGD and use of hMG were of significance in FET （ P〈 0.05）. Conclusion： A programmed cycle of BSWGD combined with low dose of hMG could improve the embryo IR and PR of FET. Embryo cryo-time, endometrial thickness, and the use of BSWGD and hMG are of significance for FET.     

阿黑皮素原相关肽对大鼠离体黄体细胞孕酮生成的影响

采用体外孵育大鼠黄体细胞的方法,观察了β-EP,β-LPH,α-MSH 和 ACTH 等阿黑皮素原(POMC)相关肽对孕酮分泌的影响,结果表明:β-EP(10~(-6),10~7,10~(-6)mol/L)可明显促进黄体细胞基础孕酮分泌,呈剂量-效应依赖关系,孕酮含量分别由对照组36.56±8.63pmol/10~6细胞上升为52.90±18.31(P<0.01),73.86±18.21和80.37±9.86pmol/10~6细胞(P<0.01);β-EP(10~(-7)mol/L)还能加强hCG 致孕酮生成作用,孕酮含量由47.30±11.57上升为63.34±9.32pmol/10~6细胞(P<0.01);纳络酮(10~(-6)mol/L)可完全阻断β-EP 的作用。与β-EP 不同,β-LPH(10~(-6)mol/L)抑制基础和 hCG 诱导的孕酮分泌;α-MSH(10~(-6)mol/L)对基础孕酮分泌无明显影响,但能加强 hCG 致孕酮生成作用;ACTH 对大鼠黄体细胞孕酮分泌无明显影响。实验显示:某些 POMC 相关肽,尤其是β-EP 可能以旁分泌的方式参与黄体细胞孕酮分泌的调节。     

醋酸棉酚对大鼠胚胎内膜及黄体的影响

本文为进一步研究醋酸棉酚对雌性大鼠抗生育机理。配后大鼠于D_1-D_4连续灌服醋酸棉酚90mg/kg,D_5回收胚胎并取子宫内膜及黄体,进行形态学观察。结果显示:D_5胚胎发育受影响,摄取~3H-TdR活性减低,偶见子宫内膜局部受损伤,内膜微血管数量明显减少;一半动物卵巢黄体细胞形态发生改变。这些因素可能是酯酸棉酚对雌性抗生育的主要环节。     

Anordrin对离体培养兔卵泡和黄体的形态学影响

本工作用多杯法培养发情兔的卵巢,观察Anordrin对家兔卵巢卵泡和黄体发育的形态变化。22只发情兔的大卵泡的培养,结果表明,在单纯培养液中加入不同剂量Anordrin处理后,卵泡生长抑制;而培养液中有FSH和LH存在时15微克/毫升或30微克/毫升Anordrin不影响各级卵泡的正常发育,Anordrin增至45微克/毫升则可使成熟卵泡的卵母细胞变性而抑制排卵。18只发情兔的黄体的培养,结果表明,无论培养液中有无FSH和LH存在,不同剂量的Anordrin均可使颗粒黄体细胞显示不同程度的变性和退化,大剂量时还可出现大量泡膜黄体细胞。以上观察表明,卵巢组织经Anordrin处理后,卵泡和黄体显示退化,反映了Anordrin对卵巢有直接的影响。     

雌/孕激素对垂体黄体生成素(LH)细胞的形态学影响

确定外源性性激素 (不同剂量雌激素、孕激素和孕激素配伍雌激素 )影响垂体黄体生成素 ( L H)分泌变化的细胞形态学特征。方法 :应用特异性 r L Hβ抗体和 ABC亲和组织化学方法显示垂体 L H细胞 ,结合计算机图像分析系统观察和测定不同剂量雌激素、孕激素和孕激素配伍雌激素对垂体 L H细胞大小、细胞内液泡大小和细胞形状等形态参数的影响。结果 :不同类型性激素使去卵巢后增大的 L H细胞恢复至正常细胞的大小范围 ,但在不同性激素影响下 ,L H细胞内最终的激素水平和相应的细胞形态特征是不同的。低剂量雌激素使 L H细胞内大量贮存的激素处于易释放状态 ,细胞内出现大小不等的液泡。大剂量雌激素抑制 L H的大量贮存 ,细胞内充满分泌颗粒 ,很少液泡结构。孕激素使细胞内激素大量贮存但处于不易释放状态 ,细胞内含有大量分泌颗粒和少量液泡结构。孕激素配伍雌激素抑制细胞内 L H的大量贮存和释放 ,但细胞形状较大剂量雌激素作用组规则 ,细胞内除分泌颗粒外也存在少量液泡。结论 :雌 /孕激素均能逆转去卵巢后 L H的过量分泌 ,并使 L H细胞呈现正常生理变化范围内的不同细胞形态变化。     

Effects of Estrogen and／or Progesterone on Morphology of Pituitary Luteinizing Hormone Cells

Objective To identify the morphological characteristics of pituitary luteinizing hormone (LH) cells after exogenous gonadal hormone treatment. Methods Effects of various doses of estrogen, progesterone and their combination on morphological parameters, including the size and shape of pituitary LH cells, the size of endocellular vacuoles, were observed and measured by immuno-histochemistry and computer image analysis. Results Different kinds of gonadal hormones could recover the magnified LH cells to the normal level in ovariectomized rats. However, their final effects on the gonadotrophin levels and the cellular morphological characters of the LH cells were different. The low dose of estrogen elicited abundant hormone stored in the LH cells to an easy released status wtth a lot oy atJJerent szze of vacuoles. On the contrary, the high dose of estrogen inhibited the storage of LH, and the LH cells were filled with secretory granules and few vacuoles. The progesterone could promote the storage of LH in an uneasy released status. The administration of estrogen-progesterone combina-tion not only inhibited the storage of LH, but also the release of LH. In this group, the LH cells containing a large amount of secretory granules and a few vacuoles showed a better uniform shape compared with those administrated with high dose of estrogen. Conclusion: Different kinds of gonadal hormones could reverse the excessive secretion of LH and recover the morphological change of LH cells to the normally physiological condition.     

小鼠窦前卵泡的体外培养

目的 观察性成熟前小鼠的窦前卵泡体外培养的形态学改变,同时评价激素及生长因子等添加物对窦前卵泡体外发育的影响。方法 机械法分离性成熟前小鼠卵巢中的窦前卵泡,分2组培养于20μL培养液滴内。(1)Gn组:α-MEM 10％FCS 100 mU/mL rFSH±ITS(5 μg/mL insulin,5 μg/mL transferrine,5 ng/mLselenium)±10 mU/mL rLH,第12天移入 α-MEM 1.5 U/mL rHCG 5 ng/mL rEGF 10％ FCS。(2)nGn组:α-MEM 10％ FCS。每天换液观察,记录GC增生、窦形成等形态学改变和卵母细胞成熟情况。结果 激素组窦前卵泡增长明显,92.92％存活,28.65％形成窦,培养14d后40.35％排出成熟卵;而无激素组窦前卵泡生长缓慢,生存6d后开始萎缩、变黑,存活率仅10.91％,无窦卵泡形成,仅11.11％排出成熟卵,差异有显著意义(P<0.01)。激素组在添加ITS、LH后,窦前卵泡的窦形成率和排出成熟卵母细胞率均显著上升(P<0.05)。结论 α-MEM可作为窦前卵泡体外培养液,但必须添加rLH/rFSH、ITS、rHCG/rEGF以促进膜细胞黏附、颗粒细胞增生和卵母细胞成熟。     

小鼠生殖泡期卵母细胞体外培养成熟、受精和胚胎发育的研究

为研究性成熟小鼠生殖泡期(GV)卵母细胞的体外成熟,同时比较体外成熟(IVM)和体内成熟组卵子的受精、胚胎发育情况,机械法分离性成熟小鼠卵巢中的GV期卵母细胞及刺激后的成熟卵母细胞,直接培养于[α-MEM+1 mg/ml Fetuin+10％FCS+1.5 IU/ml rHCG±5 ng/mlrEGF]中,16 h后移入f-HTF+10％FCS体外受精4 h,再在M16+10％FCS中培养72 h。记录其成熟、受精和胚胎发育情况,并与体内成熟的卵子比较。结果:GV期卵母细胞在含HCG培养液中成熟率为89.68％,培养液中添加EGF后,卵子成熟率无显著上升,但受精率、卵裂率均有显著提高(P<0.05)。体内成熟卵子的受精率可达62.83％,卵裂率60.18％;IVM组受精率仅48.92％,卵裂率36.69％,明显低于前者(P<0.01)。结论:小鼠GV期卵母细胞可体外培养成熟、受精和胚胎发育。培养液中添加促性腺激素和EGF都促进卵母细胞体外成熟、受精和胚胎发育能力。但IVM组卵母细胞的受精率、卵裂率低于体内成熟组,培养系统仍需改进。     

体外培养兔的卵母细胞

<正> 卵母细胞第一次成熟分裂开始是在卵巢内,卵母细胞进行成熟分裂前期只是到双线期,它的核被称为卵核(Germinal Vesicle)。早在1953年,Pincus等曾报道体外培养兔卵母细胞的卵核能在体外自发成熟,近年来许多学者已在若干种属(包括人和猴)上开展了研究。为了研究体外受精和避孕药对卵母细胞成熟的影响,我们对Pincus的方法进行了改进,建立了卵母细胞体外培养方法。