葡萄糖对ICR小鼠胚胎体外发育的影响

研究葡萄糖在小鼠早期胚胎体外发育中的作用。实验1将6—8周龄的ICR雌鼠超数排卵后与公鼠交配,收集1-细胞放入含0(对照组)、0.5、1、3、5、10mmol/L葡萄糖的CZB中培养;实验2将从超排的ICR雌鼠输卵管内收集的1-细胞放入无糖CZB中培养,分别于1细胞、2细胞、4细胞、桑椹胚阶段移入含3.0mmol/L葡萄糖(最适浓度)的CZB中,培养24h后又移回到无糖CZB中(桑椹胚阶段除外)继续培养以及整个胚胎培养过程均在含糖CZB中,对照组胚胎培养全程均在无糖CZB中。每组胚胎于37℃、5%CO2培养箱中培养120h,每24h在倒置显微镜下观察胚胎发育情况,分别计算2-细胞率、4-细胞率、桑椹胚率、囊胚率和孵化率,并进行囊胚细胞计数。结果显示,小鼠胚胎在含糖CZB中与在无糖CZB中4-细胞发育率无差异;含糖CZB中囊胚率显著高于对照组;3.0mmol/L浓度组囊胚细胞数显著高于其余组;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖囊胚率显著高于对照组,1-细胞至2-细胞、桑椹胚及其以后阶段添加葡萄糖囊胚率与对照组无差异。实验证实,在ICR小鼠胚胎体外培养中加入葡萄糖不会导致2-细胞阻滞;葡萄糖浓度增至10mmol/L对ICR小鼠胚胎无毒性作用;ICR小鼠胚胎体外培养的最适葡萄糖浓度为3.0mmol/L;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖是必要的。     

应用乙二醇冷冻小鼠胚胎：优化和简化程序的探索

提高解冻胚胎的发育能力和简化冷冻解冻程序是胚胎冷冻研究的两大永恒的主题。尽管乙二醇（EG）广泛用于家畜胚胎冷冻,但很少用于冷冻小鼠和人胚胎。为数很少的以EG慢冻小鼠或人胚胎的研究均采用较为复杂的人胚冷冻程序,未见简化程序和用EG冷冻小鼠桑椹胚的报道。采用简单的牛胚胎冷冻程序研究了发育时期、EG浓度、平衡方法、添加蔗糖以及解冻后脱除EG等对小鼠胚胎冻后发育能力的影响。结果显示：（1）致密晚期桑椹胚冻后体外培养囊胚发育率（81.92%±2.24%）和孵出率（68.56％±2.43%）显著（P＜0.05)高于4-细胞、8-细胞胚胎和致密早期桑椹胚胎;（2）1.8mol/L EG冷冻小鼠致密晚期桑椹胚的囊胚发育和孵出率显著高于其它浓度;（3）在EG中平衡10min的冻后囊胚发育显著好于平衡5、20或30min;（4）两步平衡冷冻胚胎的囊胚发育率和孵出率显著高于一步平衡;（5）用EG冷冻小鼠胚胎无需添加蔗糖;（6）解冻后可不脱除EG;（7）冻后发育的早期囊胚和囊胚细胞数明显少于体内发育胚胎。因此,用EG冷冻小鼠胚胎的最佳方案为：致密晚期桑椹胚用1.8mol/L EG不添加蔗糖、两步平衡15min、以简单的牛胚胎冷冻程序冷冻解冻、解冻后不脱除EG直接培养或移植。     

克服昆明小鼠体外受精卵发育阻滞方法的研究

本研究应用CZB和WM培养液进行昆明小鼠体外受精胚胎的发育培养,建立了一个可行的胚胎体外培养的新方法,并通过改变培养液的成分及其含量,对胚胎发育的阻滞机理和突破方法进行了初步的探索。培养于WM中的受精卵发生阻滞,有48％停留于2细胞阶段;而CZB中的胚胎有81％发育为桑椹胚和囊胚。在WM中添加EDTA和谷氨酰胺得到了66％的囊胚;加大WM中乳酸钠和丙酮酸钠的比值未能克服发育的阻滞现象。实验结果表明,EDTA和谷氨酰胺在克服阻滞时具有协同作用。     

小鼠体外受精、胚胎培养及胚胎快速冷冻的研究

目的　为扩大胚胎来源并获取特定胚龄胚胎 ,建立小鼠冷冻胚胎库。方法　运用超数排卵、体外受精与胚胎培养及胚胎冷冻技术系统研究了小鼠受精卵的体内发育与运行规律。卵母细胞的体外成熟与受精、单细胞胚胎培养及胚胎快速冷冻。结果　 (1)注射hCG后 12～ 2 0h受精卵发育至原核期 ,4 2～ 4 8h为 2 细胞期 ,4 8～ 6 0h为 4 细胞期 ,6 0～ 6 8h为 8 细胞期 ,以上各期受精卵均处于输卵管中 ;75～ 78h为桑椹胚 ,78～ 80h为致密桑椹胚 ,90～ 92h为早期囊胚 ,92～ 96h为囊胚 ,以上各期均处于子宫角中。 (2 )培养液中添加促性腺激素 (FSH与hCG) ,能显著提高卵母细胞的体外受精率 ,添加FCS和激素组的体外受精率又显著高于单独添加激素组 ,FCS还能显著提高胚胎发育。 (3)在培养液中添加EDTA ,能有效克服小鼠胚胎的 2 细胞阻断 ,其 2 细胞胚的发育率达 10 0 % ,8 细胞胚发育率达 5 5 %以上 ;牛、羊上皮细胞培养液上清也能有效克服 2 细胞阻断。添加乳酸钠和丙酮酸钠可使 2细胞与 8细胞期胚的发育率显著提高。 (4)以D PBS +甘油 +蔗糖为冷冻液 ,以D PBS +蔗糖为稀释液 ,对小鼠胚胎进行快速冷冻 ,桑椹胚的存活率为 6 9 3% ,早期囊胚的存活率为 6 0 4 %。结论　研究为将生物技术应用于小鼠 ,扩大卵子和胚胎来源     

昆明小鼠原核胚在不同培养液中的体外发育

目的优化昆明小鼠原核胚胎体外培养系统,提高胚胎发育率.方法小鼠经超排获得原核期胚胎,制备小鼠输卵管上皮共培养系统,使用M16、CZB和KSOM培养液进行体外培养,并对体内和体外发育的囊胚细胞计数.结果在KSOM和CZB中添加胎牛血清能显著提高胚胎囊胚发育率(14.71%对85.71%;6.45%对10.81%);输卵管上皮共培养可以提高胚胎的卵裂率和囊胚发育率,同时提高胚胎质量和同步发育,小鼠胚胎在KSOMFBS中囊胚发育率达85.19%,显著高于CZB和M16.结论在小鼠输卵管上皮共培养条件下,KSOMFBS能够很好支持昆明小鼠原核期胚胎体外发育.     

小鼠原核期受精卵体外培养系统的筛选

为了筛选出最佳的小鼠原核期受精卵的体外发育培养系统,分别进行了四个试验。试验I:在体外分别用自配的M16、mM16、KSOM、mKSOM、CZB进行体外发育培养,进而筛选出一种最佳的体外培养系统;试验Ⅱ、Ⅲ、Ⅳ分别:探讨了血清、PVA、rhLIF对小鼠胚胎发育的影响。结果,试验I中胚胎发育到2-细胞的比率差异不明显,但是在mM16和mKSOM中,发育到4-细胞的比率94.7%,90.7%(91/96;78/86)和发育到桑椹胚/囊胚的比率分别为78.1%,67.4%(75/96;58/86)均明显高于其他三种培养液;试验II用10?S代替M16中BSA时,胚胎的发育率均下降,即使在mM16中桑椹胚/囊胚率仅为4.8%(12/35)与对照组M16(40.5%)差异显著(p<0.05);试验Ⅲ用PVA取代mM16和mKSOM中的BSA其体外发育率显著下降,胚胎均无一例发育到桑椹胚/囊胚;试验Ⅳ:rhLIF能提高胚胎在体外的发育率可使mM16培养的胚胎囊胚率、囊胚脱出率分别达到84%(47/56)、39.2%(22/56)。结论:在不添减其他成分前提下,只在M16中添加0.1mMolEDTA、0.5mMol牛磺酸、1000IU/mlrhLIF便可获得84%的囊胚率,同时证明在M16或mM16添加血清都会降低其体外发育率;PVA还不能有效的取代mM16、mKSOM中的血清。     

小鼠囊胚的细胞凋亡:体内发育和体外培养的比较

小鼠胚体外培养到囊胚期的成功率很高,但质量是否能及体内发育的囊胚还不太清楚。细胞的数量和凋亡程度是胚胎质量鉴定的重要指标。本文采用TUNEL法分别对2-、8-细胞和桑椹胚培育成的鼠囊胚及体内发育而成的鼠囊胚细胞凋亡情况进行了检验。结果表明90%以上的2-、8-细胞及桑椹胚经过72h、48h和24h的培养发育到囊胚期。由桑椹胚发育成的与体内发育成的囊胚细胞凋亡指数没有显著差异,但由2-、8-细胞胚培育成的囊胚细胞凋亡指数显著高于体内发育成的囊胚。由此可见,体外长时间培养会增加胚胎的细胞凋亡程率。为培养出高质量的囊胚,胚胎培养条件还需进一步改善。     

人参皂甙Rg1对昆明小鼠早胚体外发育的影响

目的观察人参皂甙Rg1对昆明(kunming,KM)小鼠早胚体外发育的影响,为改善小鼠早胚体外培养体系奠定实验基础。方法以空白M16培养液为对照组,M16中添加浓度为5μmol/L、10μmol/L、20μmol/L Rg1为实验组,收集KM小鼠1-细胞胚进行体外连续培养,计数各组发育至2-、4-细胞胚、桑葚胚和囊胚等各个阶段的数目,比较各组发育至不同阶段的比率。结果添加Rg1实验组发育到桑葚胚和囊胚的比率明显高于对照组,其中以添加浓度为10μmol/L的Rg1实验组的效果最显著,其桑葚胚发育率为59.79%,而对照组只有19.17%(P<0.01)。10μmol/L的Rg1实验组囊胚发育率为17.82%,也明显高于对照组1.87%(P<0.01)。结论 Rg1可提高KM小鼠1-细胞胚体外发育到桑葚胚及囊胚的比率,以10μmol/L浓度的Rg1效果最显著。     

高浓度葡萄糖对昆明小鼠早期胚胎发育的影响

建立昆明小鼠受孕模型,分离并体外培养胚胎细胞.检测了各培养浓度下的细胞增殖、分化与凋亡.胚胎细胞在0.2mmol/L和5.56mmol/L葡萄糖浓度的KSOM培养基中能正常发育和孵化;而在浓度为15.56mmol/L和25.56mmol/L葡萄糖培养基中胚胎发育和孵化均受到损害(P<0.005),且总细胞数和内细胞团细胞数也明显减少(P<0.01),但其细胞凋亡率与0.2mmol/L和5.56mmol/L葡萄糖浓度下胚胎细胞凋亡率无显著性差异(P>0.05).随着葡萄糖浓度的增高,胚泡总的表面积无明显变化,但胚胎细胞密度呈增加趋势.高血糖对早期胚胎的发育具有毒性作用,提示高糖可能导致妊娠合并糖尿病患者的流产和胎儿畸形率升高.     

培养基中能量底物对猪胚胎发育的影响

旨在探讨丙酮酸和乳酸对猪(Susscrofa)胚胎早期发育的影响,将NCSU-23培养基中的5.56mmol/L葡萄糖替换为0.2mmol/L丙酮酸、5.7mmol/L乳酸,并将此培养基命名为mNCSU-23。根据实验设计,孤雌胚及核移植胚转移到mNCSU-23或NCSU-23中培养。激活第2天统计孤雌胚及核移植胚中的5～8细胞胚胎数。激活第6天统计孤雌胚及核移植胚囊胚形成率及囊胚细胞数。实验结果表明,mNCSU/NCSU处理组的5～8细胞胚胎数及囊胚数显著高于对照组(P0.05);单纯使用mNCSU培养猪胚胎时,囊胚率最低,发育结果最差(P0.05)。本研究证实,在体外培养前两天,用乳酸和丙酮酸代替培养基中的葡萄糖对胚胎发育有利。     

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.     

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.     

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.     

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.     

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。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.     

（鱼句）亚科花（鱼骨）型鱼类骨骼系统的比较

对我国花型Hemibarbuspattern鱼类作了骨骼系统比较,结果表明,此类型鱼类脑颅较长,副蝶骨平直或稍弯曲,眶蝶骨腹纵嵴发达(铜鱼Coreius septentrionalis例外),下颞窝和咽突中等大,基枕骨后突发达;脑颅中的上筛骨的后突、侧筛骨的外筛突,蝶耳骨的外突、上耳骨的后突、围眶骨和后颞窝等均有明显的差异;咽颅中的舌颌骨、尾舌骨、鳃盖骨和下咽齿的列数等又有显著的区别;附肢骨骼中的腰带骨、脊椎骨中的复合神经骨和第4椎骨腹侧的悬器等也有不同之处。据此,这些差异和区别可作为属间或种间的分类依据。