冻存牛体外受精胚融解后生存率与染色体分析

体外受精后 7～ 9天的牛胚胎用 1 0 %甘油 (Glycerol)冷冻保存 ,融解 48h后 ,胚胎生存率表明 ,7～ 8天的囊胚的生存率明显高于 9天的囊胚生存率 (P <0 .0 5)。染色体分析表明 ,冷冻保存 7天的囊胚卵裂球数和平均分裂像明显低于相同天数的新鲜囊胚 (P <0 .0 5)。染色体异常发生率较高 ,但与新鲜囊胚之间无明显差异。

玉米二次精胚回收工艺试验小结

“玉米干式提胚新工艺———二次精胚回收工艺” ,平均精胚提取率可达13.1% ,纯胚提取率平均达8.95% ,淀粉损失率约1.48%。新工艺结果表明 ,精胚提取率最佳值为12.7% ,纯胚提取率为8.67%。

精白保胚米发芽过程中淀粉酶及相关营养成分的变化

研究精白保胚米发芽过程中α-淀粉酶、β-淀粉酶和葡聚糖内切酶活力的变化,同时测定其中还原糖、总糖、淀粉及热水溶性直链淀粉的含量变化,并以发芽糙米作为对照。结果表明:精白保胚米和糙米发芽过程中α-淀粉酶和β-淀粉酶的活性先由低变高,再由高变低,精白保胚米在发芽16h时淀粉酶的活性最高,而糙米在发芽时淀粉酶活性达到最高点时间较长;葡聚糖内切酶活力随发芽时间的增加活性逐渐升高,糙米在发芽过程中葡聚糖内切酶活力高于精白保胚米。还原糖、总糖的含量随着时间的增加而升高,在20～24h后逐渐降低,精白保胚米在发芽过程中还原糖的含量高于糙米。淀粉含量随发芽时间的延长而逐渐降低,热水溶性直链淀粉的含量则不断增加。

精白保胚发芽米淀粉的理化性质

研究发芽对精白保胚米淀粉理化性质的影响。测定精白保胚发芽米淀粉的溶解度、膨胀度、透光率、冻融稳定性等理化性质;采用快速黏度分析仪和流变仪研究精白保胚发芽米淀粉RVA特征值的变化和流变学特性;并与糙米、发芽糙米和精白保胚米淀粉的理化性质做比较。结果表明,精白保胚米发芽之后直链淀粉含量降低,溶解度、膨胀度和透光率增加,冻融稳定性得到改善;RVA特征值表现为精白保胚发芽米的峰值黏度降低,最终黏度升高,崩解值降低,消减值和回升值升高;动态流变学表现为加热过程中精白保胚发芽米的储能模量(G')降低。

精白保胚米发芽过程中米谷蛋白及其氨基酸的变化

研究精白保胚米发芽过程中米谷蛋白及其氨基酸组成的变化。精白保胚米在发芽过程中,淀粉含量呈下降趋势,蛋白质的含量略有上升;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和体积排阻色谱的结果显示:发芽后米谷蛋白中含量较高的各亚基分子质量没有太大差别,但其含量有所变化,高分子质量亚基含量减少,低分子质量亚基含量增多。米谷蛋白氨基酸分析结果发现:发芽可以增加精白保胚米总氨基酸和必需氨基酸的含量,必需氨基酸组成模式更加合理。分析结果表明发芽可以提高精白保胚米的食用品质和营养价值。

精白保胚米发芽的工艺优化

精白保胚发芽米是一种营养丰富、富含保健功能成分的新型大米产品。通过单因素实验和响应曲面法对精白保胚米发芽的浸泡、发芽温度和时间及催芽方法进行了优化,获得精白保胚米发芽的最佳工艺为:在温度25℃条件下以0.5%的饱和碳酸钙溶液浸泡17h,再以超声波催芽10min,置于相对湿度90%、温度25℃的条件下发芽21h,在此条件下,发芽率可达94%以上。实验结果为精白保胚发芽米的研制和推广奠定了基础。

发芽对精白保胚米淀粉分子结构的影响

目的:研究发芽对精白保胚米淀粉分子结构的影响。方法:采用扫描电子显微镜、高效液相色谱、X射线衍射仪及傅里叶红外光谱(FT-IR)分析精白保胚发芽米淀粉颗粒的表面结构、相对分子质量分布、结晶度和特征结构的变化。结果:精白保胚米发芽之后淀粉颗粒表面结构变化不明显,但淀粉的相对分子质量、结晶度降低;发芽后精白保胚米淀粉的红外透光率降低,O-H键、C-H键和C-O键的伸缩振动均增强。结论:发芽过程中直链淀粉和支链淀粉降解,使精白保胚米淀粉分子结构发生明显变化。

精白保胚发芽米食用品质

以糙米、发芽糙米、精白保胚米及精白保胚发芽米为研究对象,对其食用品质的主要方面,即炊饭方法、蒸煮特性、糊化特性、质构特性及感官分析进行对比评价。结果表明:精白保胚发芽米的吸水率和膨胀率均低于其他3种大米,而米汤固体溶出物和碘蓝值均高于糙米和发芽糙米;糊化特性分析中,精白保胚发芽米与大米食用品质相关的崩解值降低,消减值升高;质构分析表明精白保胚发芽米具有最低的硬度;感官分析结果表明,精白保胚发芽米在色泽、香气、口感、滋味方面都优于未发芽米。

Changes of Calmodulin Distribution in the Embryo Sac of Oryza sativa Before and After Fertilization: an Immunogold Electron Microscope Study

Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.

Embryonic development of the concave-eared torrent frog with its significance on taxonomy

We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota （Amphibia, Anura, Ranidae）. Embryos were derived from artificial fertilization of frogs’ eggs, and the staging of development was based on morphological and physiological characteristics. Two major periods of development were designated： i） early embryonic period, from fertilization to operculum completion stage, lasted for 324 h at water temperature （WT） 18 ？23℃; ii） larval period, from operculum completion stage to tail absorbed stage, took 1207 h at WT 20 ？ 24℃. Tadpoles of the concave-eared torrent frogs showed no evidence of abdominal sucker. Absence of this key characteristic supports the view from molecular systematics that concave-eared torrent frog does not belong to the genus Amolops. Two cleavage patterns were observed in embryos at 8-cell and 16-cell stages, with Pattern I2 （latitudinal cleavage at the 8-cell stage, and meridional cleavage at the 16-cell stage with two perpendicular meridional furrows） being the predominant pattern and only 1.5% belonging to Pattern II （meridional cleavage at the 8-cell stage and latitudinal cleavage at the 16-cell stage）. The factors affecting cleavage and hatching ratios, developmental speed, and ecological adaptation were discussed.

Embryogenesis of Polyembryonic Rice ApⅢ: Structural and Histochemical Studies of Egg Apparatus Around Fertilization

The structural and histochemical changes of the egg apparatus in the polyembryonic rice ( Oryza sativa L.), ApⅢ with the highest frequence of additional embryos among the polyembryonic rice investigated, before and after fertilization were studied and compared with those of normal and other polyembryonic rices in a similar developmental period. A total of 2 932 ovules were observed and each of them contained only a single embryo sac with a set of egg apparatus. Among 1 655 embryo sacs, there were 1 643 embryo sacs (99.27%) with one normal egg apparatus in each embryo sac, and only 12 embryo sacs (0.73%) from the remainder with 4_celled egg apparatus, i.e. two eggs and two synergids. Neither the numerous poly_egg apparatus and egg_like cells, nor the double set of embryo sacs each containing one egg apparatus and other abnormal egg apparatus in single ovary, which were reported by earlier investigators to have high frequency of embryo production in SB_1 and ApⅣ, were observed. The egg cell was located at the subterminal site of the micropylar end of embryo sac. The cytoplasm of egg cell was rich in protein materials and polysaccharide grains, which did not disappear until the division of zygote. The prominent nucleus was closely surrounded by protein and polysaccharide grains, which did not disappear until the division of zygote. No cytological difference was found between egg cells from the normal and abnormal egg apparatus. The two synergids were fully developed and situated at the upper most part of the micropylar end of the mature embryo sac. In most embryo sacs, the synergids were flask_shaped with longer necks, and a widened cap_shaped top, in close contact with the micropyle. The synergids had a well developed filiform apparatus. The characteristic appearance of the filiform apparatus as well as the cap_neck region of synergids before and after pollen tube penetration were easily distinguishable from the egg cell. The structure, the stainability with Coomassie Brilliant Blue and PAS reaction, the process of accumulation, distribution and disapperance of the cytoplasmic protein materials and polysaccharide grains of the two synergids, the persistent and rarely the receptive synergids before and after pollen tube penetration, were closely similar to those of egg cell of the same developmental stage. In comparison with normal and other polyembryonic rice reported, the size of nucleus and nucleolus and their stainability also strongly resembled those of egg cell. Based on the results observed, the main conclusions are summarized as follows: (1) the additional embryos very frequently developed in the young and mature seed of polyembryonic rice ApⅢ were produced by one or two synergids of normal egg apparatus, rarely by 4_celled egg apparatus; (2) during fertilization, the synergids, in addition to the natural specific function of introducing pollen tube and transferring sperms to egg cell and central cell, could be closely associated with the potentiality to breed one or two additional embryos; and (3) as compared with that of normal or other polyembryonic rice it is firstly disclosed that in a few embryo sacs of ApⅢ, the cytoplasmic and nuclear structure, the active anabolism and catabolism of protein and polysaccharide materials and the delayed disorganization at the mid_basal region of the receptive and persistent synergid still remained unchanged before the division of zygote. Such salient features could be the predisposition for the origin of additional embryos in ApⅢ.

Cytological Mechanism of Cytoplasmic Inheritance in Pinus tabulaeformis: Ⅱ. Transmission of Male and Female Organelles During Fertilization and Proembryo Development

In an earlier report the ultrastructure and nucleoid organelles of male gamete in Pinus tabulaeformis Carr. have been described. Presently, the ultrastructure of the cytoplasm of the egg cell and pollen tube—immediately before fertilization and during cytoplasmic transmission of male gametophyte—has been described for the same species. The fate of parental plastids and mitochondria in the proembryo has also been followed. The mature egg cell contains a large amount of mitochondria, but seems to lack normal plastids. Most plastids have transformed into large inclusions. Apart from the large inclusions, there are abundant small inclusions and other organelles in the egg cell. During fertilization, pollen tube penetrates into the egg cell at the micropylar end and thereafter the contents are released. Plastid and mitochondrion of male origin are lacking near the fusing sperm_egg nuclei. The second sperm nucleus—not involved in karyogamy—remains at a site near the receptive vacuole. This nucleus is surrounded by large amount of male cytoplasm containing mixed organelles from the sperm cell, tube cell, and egg cell. At the free nuclear proembryo stage, organelles of male and female origin are visible in the perinucleus_cytoplasmic zone. Most of the mitochondria have the same morphological features as those in the egg cell. Some of the mitochondria appear to have originated from the sperm and tube cells. Plastids are most likely of male gametophyte origin because they have similar appearance as those of the sperm and tube cell. Large inclusions in the egg cell become vacuole_like. Paternal plastids have been incorporated into the neocytoplasm of the proembryo. In the cellular proembryo, maternal mitochondria are more abundant. Plastids resembling those of the sperm and tube cell are still present. These cytological results clearly show that in P. tabulaeformis, plastids are inherited paternally and mitochondria bipaternally. The cytological mechanism of plastid and mitochondrion inheritance in gymnosperm is discussed.

Research on Embryonic Development of Loach

[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5. The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage. The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] It provided important reference for studying artificial propagation and genetic breeding of loach.

Study on Artificial Propagation and Embryonic Development of Poyang Lake Catfish

Poyang Lake catfish （Silurus asotus Linnaeus） is native to Poyang Lake Basin. Artificial propagation of Poyang Lake catfish was carried out by using artificial insemination. Under water temperature of 18-26 ℃, embryonic development of Poyang Lake catfish in impounded water was observed and recorded in detail. Embryonic development of Poyang Lake catfish encountered seven stages, and it last- ed for 47.08 h from the fertilized egg stage to the stripping stage. The hatching time of Silurus meridionalis Chen was shortest, followed by that of Poyang Lake catfish, and the hatching time of Pelteobagrus vachelli was longest.

The Study on the Gene Expression of Preimplantation IVF Bovine Embryos

The cattle different stage embryos obtained from in vitro was studied using the technology of single preimplantation embryo mRNA different display:single 8-cell and blastocyst stage embryos were studied using technology of mRNA different display and one different fragment was found. The result suggested that this fragment displayed high homology (99%) to cattle mRNA for ribosomal protein L31. Then to detect the expression of RPL31mRNA in 8 cell and blastocyst stage embryos by real-time quantitative PCR,the result showed the relative amount of 8 cells was 3.2 times of blastocyst's.

Accuracy of a combined score of zygote and embryo morphology for selecting the best embryos for IVF

Objective:To evaluate the accuracy of a scoring system combining zygote and embryo morphology in predicting the outcome of in vitro fertilization (IVF) treatment. Methods: In a study group, 117 consecutive IVF or intracytoplasmic sperm injection (ICSI) cycles with embryo transfer were carried out and 312 embryos were scored using a combined scoring system (CSS) of zygote and embryo morphology before transplantation. In a control group, a total of 420 IVF or ICSI cycles were carried out and 1176 embryos were scored using a cumulative embryo score (CES). The effects of the combined scoring system on the embryo implantation rate and pregnancy rate per cycle were analyzed. Results: Using the combined scoring system, the embryo implan-tation rate (27.6%) and the clinical pregnancy rate (48.7%) were significantly higher than those in the control group (20.8% and 38.6%, respectively). Also, the implantation rate of embryos scoring ≥70 (38.5%: 82 sacs/213 embryos) was significantly higher (P<0.001) than that of embryos scoring <70 (4%: 4 sacs/99 embryos). The pregnancy rate of patients with embryos scoring ≥70 using the combined scoring system (66.7%) was significantly higher (P<0.001) than that of patients with embryos scoring ≥20 using the cumulative embryo score (59.0%). Conclusion: The results suggest that selecting embryos with a high score (≥70) using the combined scoring system could increase the implantation rate and pregnancy rate, and that using a scoring system combining assessments of human zygotes and pre-implantation embryos might predict IVF outcomes more accurately than using a cumula-tive embryo score.

Gastric adenocarcinoma with features of endodermal sinus tumor

Extragonadal germ cell tumors are rare. The most common sites for EGGCTs are in midline locations such as the mediastinum, retroperitoneum and pineal gland. These tumors rarely present in the stomach. We describe here a case where a middle aged man presented with typical symptoms of gastric cancer. After extensive workup, which included blood work, CT abdomen scan, upper endoscopy, and endoscopic ultrasound, the patient was diagnosed with gastric cancer. However, due to very high blood levels of alpha-fetoprotein, the specimen was sent for special histochemical staining, which demonstrated that the tumor had features of both adenocarcinoma and endodermal sinus tumor. This is a very aggressive tumor with a very poor prognosis.

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFT-EYED FLOUNDER, PARALICHTHYS OLIVACEUS (T. et S.)

The early effects of chromosomal manipulation of eggs and sperm on the yields of triploid and gynogenetic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fertilized eggs at 0-2℃ for 45 minutes duration 5 minutes after fertilization, and the induced triploidy rates were 31.2%-50% and the relative hatching rates were 53.3%-99%. Gynogenetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0-2℃ for 45 minutes duration 5 minutes after fertilization. The induced gynogenetic diploid rates and the relative hatching rates were 94%-96% and 48.5%-68.5% respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae according to Student’s t test (α=0.05). The relationship between the early growth of the triploid experimental group larvae and that of gynogenetic diploid experimental group larvae was also studied.

Epigenetic effects of ethanol on liver and gastrointestinal injury

Alcohol consumption causes cellular injury. Recent developments indicate that ethanol induces epigenetic alterations, particularly acetylation, methylation of histones, and hypo- and hypermethylation of DNA. This has opened up a new area of interest in ethanol research and is providing novel insight into actions of ethanol at the nucleosomal level in relation to gene expression and patho-physiological consequences. The epigenetic effects are mainly attributable to ethanol metabolic stress (Emess), generated by the oxidative and non-oxidative metabolism of ethanol, and dysregulation of methionine metabolism. Epigenetic changes are important in ethanol-induced hepatic steatosis, fibrosis, carcinoma and gastrointestinal injury. This editorial highlights these new advances and its future potential.

Role of the first mitosis in the remodeling of the parental genomes in mouse embryos

Although male and female pronuclei reside in the same zygotic cytoplasm, they differ in many respects, such asvolume and transcriptional activity. The aim of this study is to investigate whether these differences are lost during thefirst mitosis. For this purpose, a new method was developed to inhibit the mixing of two parental chromosomes duringmitosis, thus to induce the formation of two nuclei after they exit from the mitotic phase. In this method, one-cellembryos are arrested at metaphase by treatment with nocodazole, and whn exitting from the mitotic phase, two nucleiwere formed in a single karyocyte following treatment with 6-dimethylaminopurine (6-DMAP). These embryos weredesignated as post-mitotic embryos (PM-embryos), in which the two nuclei were derived from the male and femalegenomes. We found that in the control one-cell embryos that had not been treated with the reagents, the volume of themale pronucleus was about 1.65-fold greater than that of the female pronucleus, whereas the volumes of the two nucleiin the PM-embryos were similar (volume ratio of 1.01). Although a two-fold difference in transcriptional activity wasdetected between the male and female pronuclei in the control embryos, no difference in transcriptional activity wasdetected between the two nuclei of PM-embryos. The ratio of transcriptional activity in the nucleus derived from thepaternal genome to that from the maternal genome was 1.02, for which no significant difference was detected by the χ2fitness test. Therefore, the volumes and transcriptional activities of the male and female nuclei were approximately equalin PM-embryos, which suggests that the asymmetries of pronuclear volume and transcriptional activity between maleand female genomes are somehow losted during the first mitosis.