极严重少弱精症患者的新鲜周期移植中卵裂球数目对妊娠结局的影响

目的:探讨极严重少弱精症患者的新鲜周期移植中卵裂球数目对妊娠结局的影响。方法:回顾性分析392个新鲜胚胎移植周期,这些胚胎根据移植天数分为第二天移植(Day2)组68周期和第三天移植(Day3)组324周期,其中Day2组根据卵裂球数分为3个(A组,3周期),3～5个(B组,57周期),5个(C组,8周期),Day3组根据卵裂球数分为7个(D组,33周期),7～9个(E组,242周期),9个(F组,49周期)。每个组又根据胚胎评级分为三个亚组,亚组1均为移植1～2级的胚胎(A1～F1),亚组2均为移植3～4级的胚胎(A2～F2),亚组3为一个1～2级和一个3～4级移植胚胎(A3～F3)。综合比较各组和相应亚组之间的种植率,临床妊娠率,流产率及活产率的差别。结果:①在day2组中,ABC三组的胚胎种植率,临床妊娠率,流产率及活产率均无统计学差异(P0.05)。在Day3组中,E组的胚胎种植率,临床妊娠率及活产率均高于D组,差异具有统计学意义(P0.05)。而D组和F组及E组和F组之间差异均无统计学意义。②在相同卵裂球数组中,不同的胚胎分级各亚组之间(如B1,B2,B3间)的胚胎种植率,临床妊娠率,流产率及活产率均无统计学差异。不同卵裂球数其相应的同一亚组之间(A1,B1,C1等)上述妊娠结局指标无统计学差异。结论:严重少弱精形成的胚胎优先推荐第二天卵裂球数为3～5个及第三天卵裂球数为7～9个的优质胚胎进行移植。     

实验兔胞内单精子注射技术

目的 建立实验兔胞内单精子注射技术(intracytoplasmic sperm injection,ICSI).方法 实验1比较了hCG注射后不同取卵时间对ICSI胚体外发育的影响.实验 2 比较了不同的激活方式对ICSI胚体外发育的影响.实验 3 比较了不同状态的兔精子ICSI胚胎体外发育结果.结果 (1)hCG注射后14 h取卵,其卵裂率、桑椹胚率和囊胚率(82.2%、72.9%和62.2%)都比16 h(75.9%、70.0%和53.3%)的高,但是差异无显著性(P>0.05);18 h取的卵注射后不能卵裂.(2)机械刺激组和离子霉素 6-DMAP组,ICSI后其卵裂率分别为82.2%和81.1%(P>0.05),桑椹胚率分别为72.9%和66.2%(P>0.05),囊胚率分别为51.3%和62.3%(P<0.05),机械刺激组和离子霉素组之间卵裂率和桑椹胚率差异无显著性,但是囊胚率差异有显著性.(3)新鲜精子组和冻融活精子组卵裂率(81.1%和68.8%)和囊胚率(62.3%和40.4%)差异有显著性(P<0.05),而桑椹胚率(66.2%和61.9%)差异无显著性(P>0.05).结论 精子冷冻前后,通过ICSI所得的桑椹胚均能孵化,表明已初步建立了实验兔的ICSI技术.     

初次卵裂时间是猪克隆胚胎发育潜能的重要标识

初次卵裂时间与哺乳动物胚胎发育潜能有关．比较了不同初次卵裂时间(20～24 h,早期;25～36 h,中期;37～48 h,晚期;20～48 h,对照)的猪孤雌(parthenogenetic,PA)、体细胞核移植(somatic cell nuclear transfer,SCNT)胚胎的囊胚发育率、扩张囊胚发育率和囊胚细胞数,评价其体外发育能力．发现早期卵裂的PA胚胎发育到第6天的囊胚发育率显著高于中期、晚期以及对照组(P < 0.05;54.0% vs. 19.6%,5.4%,18.7%)．扩张囊胚发育率,早裂胚胎同样优于其他组．早期卵裂的SCNT胚胎发育到第6天的囊胚比率高于中期卵裂胚胎(32.2% vs. 23.5%),而晚期卵裂胚胎发育到囊胚的比率最低(6.3%)．早期卵裂的SCNT胚胎发育到第6天的扩张囊胚比率显著高于其余各组 (P < 0.05;18.9% vs. 5.9%、3.1%、7.4%)．囊胚细胞数在早期、中期、晚期三组之间表现出下降趋势．将早期卵裂的SCNT胚胎与未经挑选的对照组胚胎分别进行移植,观察其体内发育能力．移植早裂SCNT胚胎的受体在产仔数和克隆效率上均明显高于未经挑选胚胎的受体(4.7 vs. 2.1;3.9% vs. 0.9%),说明早裂胚胎着床后具有更强的发育能力．以上结果表明：初次卵裂时间可以作为猪克隆胚胎发育潜能的重要标识,选择早裂的胚胎进行移植,有助于提高克隆效率．     

绵羊胞内单精子注射技术

本文的主要目标是建立绵羊胞内单精子注射技术(intracyioplasmic sperm injection,ICSI)。并且尝试了ICSI技术生产绵羊转基因胚胎的可能性。实验1比较了绵羊卵母细胞孤雌激活的3种化学方法。结果显示,Ionomvcin/3 h/6-dimethylaminopurine(6-DMAP)和Ionomycin激活胚的卵裂率(71.7%和91.8%)和囊胚率(17.4%和11%)显著(P<0.01)高于Ca~(2+)激活胚(18.4%和0)。Ionomycin/3 h/6-DMAP激活胚的囊胚发育形态和比例相对较高。实验2中,活精子注射至卵母细胞质后,用Ionomycin/3 h/6-DMAP激活,排出第二极体(PB2)的71枚ICSI胚胎用SOFaaBSA溶液培养,卵裂率为71.8%(51/71),显著(P<0.01)高于体外受精(41.4%,IVF)和阴性对照胚胎(30.2%,sham-ICSI)。培养7天后,sham-ICSI组没有囊胚生成;ICSI和IVF胚胎的囊胚率分别为7.0%(5/71)和16.1%(9/56),两者差异不显著(P>0.05)。这些ICSI囊胚在冷冻前后均能孵化,显示初步建立了绵羊ICSI技术。另外,我们探索了ICSI技术生产转基因胚胎的可能性,-20℃冻融1次的死精子与pEGFP-N1质粒共注射,33枚2-细胞期ICSI胚胎中,2枚可见GFP蛋白。其中1枚停止发育,另外1枚继续发育至16-细胞期,仍然可见GFP基因的表达。4枚解冻的ICSI囊胚手术移植给2只发情同期化受体绵羊,60天时,B超未见怀孕。本文初步结果表明,ICSI技术生产转基因绵羊有可能性,需深入研究。     

正常和冷冻保存复苏后的卵裂期胚胎的体外囊胚形成能力的观察

由囊胚中获得ICM是胚胎干细胞研究的基础,本文对体外授精/胚胎移植（IVF-ET）中移植后剩余的胚胎和冷冻复苏后胚胎的囊胚形成能力进行了观察。结果表明：冷冻复苏胚胎,培养3d移植后剩余胚胎,连续培养4-6d的胚胎及培养6d后的剩余胚胎的囊胚形成率分别为11.8%(6/51),28.3%(36/127),42.0%(74/176)和9.5%(2/21)。各组结果均显示卵裂细胞的正常卵裂时相的发育状态直接决定其囊胚形成能力。认为由剩余胚胎和冷冻保存复苏后的胚胎中获得囊胚是可行的。     

激光辅助孵化对冻融第三天胚胎囊胚培养后移植结局的影响

目的:探讨实施激光辅助孵化对冻融的第三天胚胎进行囊胚培养后移植妊娠结局的影响。方法:回顾性分析冻融第三天胚胎行囊胚培养和移植的542例患者的临床资料,其中164例接受激光辅助孵化(LAH组),378例没有进行激光辅助孵化(NLAH组),比较两组患者的胚胎种植率、着床率、临床妊娠率及流产率。结果:LAH组和NLAH组患者的年龄、不孕年限、体重指数、解冻胚胎数目、存活胚胎数目、移植胚胎数目比较均没有显著性差异(P0.05),LAH组的临床妊娠率高于NLAH组,流产率低于NLAH组,但差异无统计学意义(P0.05)。LAH组中≤30岁患者的着床率和临床妊娠率均高于NLAH组中≤30岁患者,而流产率则较低;LAH组≤30岁,30~34岁以及34岁患者的流产率均分别低于NLAH组,但差异均无统计学意义(P0.05)。结论:激光辅助孵化并没有明显改善冻融第三天胚胎囊胚培养后移植的临床结局。     

家兔供体卵裂球细胞周期对核移植胚胎发育的影响

通过化学药物处理使家兔桑椹胚卵裂球的细胞周期分别同期化于G1、S或G2期 ,然后分别移入去核的MⅡ期卵母细胞中 ,以研究供体核细胞周期对家兔胚胎细胞核移植效率的影响。试验结果表明供体核细胞周期对家兔核移植胚胎的发育潜力有明显影响 ,以G1期卵裂球为供体的核移植重组胚的激活率、卵裂率、桑椹胚、囊胚和孵化囊胚率及囊胚平均细胞数分别为 87 3% (32 2 / 36 9)、 84 9% (2 99/ 35 2 )、 71 5 % (193/ 2 70 )、5 8 0 % (76 / 131)、 35 1% (4 6 / 131)和 12 6± 4 8,显著高于S期 [79 6 % (10 9/ 137)、 74 4% (93/ 12 5 )、30 3% (33/ 10 9)、19 3% (17/ 88)、 6 8% (6 / 88)和 118 8± 3 5 ]、G2期 [6 3 6 % (70 / 110 )、6 0 % (6 0 / 10 0 )、16 9% (11/ 6 5 )、 16 3% (7/ 43)、 4 7% (2 / 43)和 110 6± 5 8]和未经同期化处理的卵裂球 [78 1% (185 /2 37)、 73 2 % (15 8/ 2 16 )、 49 4% (4 3/ 87)、 32 2 % (2 8/ 87)、 12 6 % (11/ 87)和 12 0 5± 4 4](P <0 0 5 )。来源于G1期卵裂球的 144枚克隆胚胎移植到 12只受体中 ,6只妊娠并产下 19只活仔 ,产仔率为 13 2 % ,显著高于来源于S期 (5 8% ,7/ 12 0 )、G2期 (0 ,0 / 12 6 )或未同期化卵裂球的克隆胚胎 (5 3% ,8/ 15 0 ) (P <0     

冷冻胚胎移植周期中内膜形态及内膜厚度对妊娠结局的预测价值分析

目的:评价内膜厚度及内膜形态对于体外受精-胚胎移植中冷冻胚胎解冻复苏移植的临床妊娠结局的预测价值。方法:回顾性分析1521个冷冻胚胎解冻复苏移植周期,将患者按子宫内膜厚度分为4组,子宫内膜≤6 mm、6.1-8.0 mm、8.0-12 mm、12.0 mm,根据子宫内膜形态分为A型内膜、B型内膜、C型内膜。分别比较不同内膜厚度分组及不同内膜形态分组患者的年龄,移植胚胎数目、内膜准备方案构成比、移植胚胎类型(卵裂期胚胎、囊胚)构成比及各组间的临床妊娠率和活产率。采用ROC曲线分析子宫内膜厚度、形态对临床妊娠结局的预测价值。使用逐步回归分析内膜厚度、形态、年龄及移植胚胎类型与妊娠结局的相关性。结果:纳入1521个解冻复苏周期中按内膜厚度分为4组,各组周期数分别为96周期、454周期、893周期、78周期,各组间平均年龄分别为34.1±5.5岁、33.3±5.4岁、32.5±5.2岁、33.7±6.0岁(P0.05)。内膜厚度≤6 mm组临床妊娠率为31.3%,子宫内膜≤6mm、6.1-8.0 mm、8.0-12 mm、12.0 mm组临床妊娠率分别为48.5%、51.8%、47.4%(P0.05)。子宫内膜≤6 mm、6.1-8.0 mm、8.0-12 mm、12.0 mm组间活产率分别为18.8%、37.7%、44.6%、39.7%(P0.05)。A型、B型及C型内膜组周期数分别为920周期、189周期及412周期,三组间平均年龄分别为32.3±5.1、33.0±5.7、34.2±5.5岁(P0.05)。A型/B型及C型内膜组临床妊娠率分别为51.2%、46.6%及46.4%,三组间活产率分别为42.1%、36.0%及37.1%,三组间差异无统计学意义(P0.05)。子宫内膜厚度ROC曲线下面积为0.534(95%可信区间0.505-0.564),子宫内膜形态ROC曲线下面积为0.526(0.476-0.955)。逐步回归分析纳入内膜厚度、内膜形态、年龄、胚胎类型4个可变量,女方年龄(OR=0.929,P0.001),移植胚胎中囊胚比例(OR=1.595,P0.001)与临床妊娠率明显相关,内膜厚度(OR=1.054,P=0.05)及内膜形态(OR=0.864)与临床妊娠率无相关性。结论:虽然子宫内膜薄临床妊娠率下降,但是子宫内膜厚度与内膜形态不能够预测体外受精-胚胎移植解冻复苏周期临床妊娠率,患者年龄以及移植胚胎的发育潜能才是预测临床妊娠率的参考指标。     

小鼠2-细胞胚胎单个卵裂球体外培养及其发育形成囊胚的玻璃化冷冻保存技术的研究

单个卵裂球分离和培养是生产同卵双胎或一卵多胎动物的一种有效方法。在羊和牛已经获得了来源于2-细胞和4-细胞期胚胎的单个卵裂球的活体后代;在兔也获得来源于4-细胞胚胎的单个卵裂球后代;在猪已获得8-细胞单个卵裂球的后代。但多数的研究者发现,小鼠单个卵裂球培养后的体外发育率、移植妊娠率和产仔率都很低。上述均为新鲜胚移植结果,至于分离后卵裂球发育成的囊胚再进行玻璃化冷冻保存尚未见报道。本实     

评价两种消毒剂对IVF实验室微环境和妊娠结局的影响

目的在辅助生殖体外受精(in vitro fertilization,IVF)实验室的胚胎操作过程中,根据消毒剂对胚胎质量和妊娠结局的影响,评价季铵盐消毒剂是否比75%酒精消毒液更适合用于IVF实验室。方法回顾分析2016年1月至9月在我中心行IVF的619例新鲜移植周期,根据IVF实验室使用的消毒剂不同分为两组,A组383例为使用75%酒精消毒组,B组236例使用季铵盐组。对比两组的VOC浓度、获卵数、获卵率、受精率、多精率、卵裂率、优质胚胎率、优质囊胚形成率、着床率、临床妊娠率以及流产率。结果相比于A组,B组的VOC浓度显著降低;A、B两组的获卵数、获卵率、受精率、多精率、卵裂率、优质胚胎率、优质囊胚形成率和流产率的差异均不具有统计学意义(P0.05);但B组的着床率和妊娠率显著高于A组(P0.05)。结论两组对胚胎的体外培养质量没有显著性的影响,但使用季铵盐组相比于75%酒精组胚胎着床率和妊娠率显著提高。因此,季铵盐消毒剂比75%酒精更适合作为IVF实验室消毒剂。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.