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成熟作用

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Japonicus and analyzed the role of animal process of oocyte during ovulation and oocyte maturation. What's more, we studied the role of microtubules and microtubule organizing center upon GV migration. The oocyte of A. japonicus connects to the follicle via animal pole process. When placed in normal sea water, ovulation started spontaneously.

利用DTT诱导刺参卵母细胞成熟,综合运用光镜、电子显微镜及激光共聚焦技术观察卵母细胞成熟过程,分析了卵母细胞表面动物极突起在卵母细胞脱滤泡中的作用,以及卵母细胞动物极突起和微管组织在生发泡移动过程中所起的作用。

By immunohistochemistry and in situ hybridization, GCNF was regionally expressed in the corpus, with no expression or weak expression in caput and cauda.

GCNF在附睾中的特异表达,表明GCNF可能参与精子在附睾中的成熟作用

The expression level of metabolitic enzyme (CYP1A1, CYP1B1) and its correlation with cell differentiation /apoptosis were studied as well. The influence of resveratrol in cell cycle was determined with FCM. Results 1 Resveratrol not only suppresses the growth of medulloblastoma cells in a dose and time related fashion but also induces cell differentiation and apoptosis. Resveratrol promotes differention of UW228-1,-2 cells to forward glia, UW228-3 and Med-3 cells to neuron. A treatment of resveratrol in the concentration of 100μM for 48hrs comit most of cells die of apoptosis. 2 Among the four cell lines Fas and Caspase-3 were constantly expessed, whereas FasL was absent irrespective to the drug treatment. 3 ICC, RT-PCR, Western-blot hybridization and EROD enzymatic activity assay demonstrated that expression of CYP1A1 is different after treatment with resveratrol in four cell lines, up-regulated in three UW228 cell lines in a dose dependent manner but down-regulated in Med-3 cells. Suppressive effect of resveratrol on CYP1B1 expression is the same among four medulloblatoma cell lines. 4 Cell cycle distribution of UW228-3 was greatly changed after treatment.

结果:1、白藜芦醇以时间和剂量依赖性方式抑制髓母细胞瘤细胞生长,同时促进细胞的分化和凋亡:白藜芦醇促使UW228-1、-2靶细胞向神经胶质细胞方向成熟分化,UW228-3、Med-3细胞向神经元细胞方向成熟分化,以100μM作用48小时效果显著;2、四个细胞系均有Fas和Caspase-3表达,但无Fas-L基因表达和蛋白活性产生,因此难以形成Fas相关性死亡通路;3、白藜芦醇处理前后CYP1A1基因的表达在各细胞系略有不同:UW228-1,-2,-3三个细胞系白藜芦醇以剂量相关性方式上调CYP1A1的表达,而Med-3细胞系,白藜芦醇却抑制CYP1A1的表达;四个细胞系在白藜芦醇处理后CYP1B1的表达均受到抑制;4、UW228-3细胞系经白藜芦醇作用后细胞周期有较大改变,表现在G0/G1期在整个细胞周期中所占的比例增加,而其它时期则相应减少。

Vg could be detected in haemolmph of the water—fed female at 48h after eclosion, too.

用10%的蜂蜜和清水分别饲喂两组成虫(简称"蜜组"和"水组"),以研究成虫期的营养对Vg发生和卵巢发育的影响,结果显示,成虫期营养对于Vg的启动合成并不起关键的决定性作用,两组成虫均可以在羽化的48小时检测到Vg,且水组中的血淋巴水平并不低于蜜组对照,但绝大多数水组个体的卵巢发育停留在卵黄沉积的某一时期,此时在血淋巴中也检测不到,只有约20%的饲水个体卵巢可以发育成熟,出少量卵;用高剂量的ZR515于羽化当日点滴处理饲水个体,可以使所有接受处理的饲水个体的卵巢发育成熟,但是产卵〓依旧低于对照组的,不能恢复正常;这说明成虫期营养对卵巢中全体卵母细胞的成熟是必需的,其作用的产生是通过内分泌系统途径实现的。

Changes of the activity of ACC oxidase were coincident with that of ethylene production and ABA content.

乙烯在跃变型果实成熟中的作用已被普遍认可[2],而脱落酸在非跃变型果实的成熟过程中起关键作用[3 4]。

DCs acquired by our reformed methods express both CD83 and CD 14 molecules highly, and have a higher density than other domestic reports. The higher TNF in DCs culture medium of HC patient suggests DCs in patient still have antigen presenting ability and by optimization the culture medium would improve its presenting ability and have a potential value in design and application individual vaccine. Although antigens pulsed DCs have a decrescent antigen presenting ability but BCG HSP70 could induce its mature and improve its presenting ability. Suggests BCG HSP70 would be a useful mature inducer. Lymphocytes primed by DCs based HC vaccine have the specific cytotoxicity against HCC lines. The CTL after freezing and anabiotic could prophylaxis and therapy HC xenograft on nude mouse. The results also suggests that CD4〓 lymphocytes play a important role in HC with a good differentiation and would be useful in treatment this kind of HC. After being activated by Peptide LLNQHACAV of hAFP and apoptotic HCCs pulsed DCs respectively, the culture medium of activated lymphocytes both contains a high level Th1 cytokines IL-12 and TNF. Primed lymphocytes appeared a characteristic of NK cells. DCs not only inhibited the growth of human HCC and other cancer cells in vitro but also prevented the growth of HC xenograft on nude mouse in vivo. There are at least three kinds of mechanism playing important role in DC based vaccine ,there are inhibition of DCs, HC specific CTL and cytokines pathway.

诱导出的DC共同表达CD83和CD14分子,CD83分子表达明显高于国内报道;肝癌患者DC培养上清中TNF水平高于健康人,提示肝癌患者DC仍具一定的抗原呈递能力,适当调控可使其行使APC功能,以期在肝癌个体化疫苗中发挥作用;DC负载肝癌可溶性抗原后,抗原呈递能力降低,BCG HSP70可促进DC成熟,增加其抗原呈递能力,预示BCG HSP70有可能成为促进DC活化和成熟的另一重要分子;肝癌DC疫苗在体外诱导肝癌特异性淋巴细胞,活化的淋巴细胞在体外对肝癌细胞的杀伤以特异性CTL为主,同时分泌较高水平Th1型细胞因子IL-12和TNF,并抑制4种肝癌细胞生长;冷冻复苏后的肝癌特异性淋巴细胞可以预防和抑制人肝癌裸鼠皮下移植瘤,提示DC负载肝癌可溶性抗原后诱发的MHC-Ⅱ类限制性CD4〓T细胞有可能在分化程度高的肝癌治疗中发挥作用;用DC和HLA-A2〓DC分别负载凋亡肝癌细胞和hAFP218-226位LLNQHACAV HLA-A2限制性九肽,在体外诱导肝癌特异性淋巴细胞,活化后的CTL细胞分泌较高水平的Th1型细胞因子IL-12和TNF,并具较强杀伤活性,此CTL同时具备NK细胞特征;DC对肿瘤细胞的抑制作用可能是通过吞噬实现的,Fas-L在DC抑制中也起一定作用;DC对人肝癌裸鼠皮下移植瘤的抑制率为97%;在肝癌DC疫苗的作用中,至少联合3种以上机制,即通过DC的直接作用、肝癌特异性CTL和细胞因子途径直接或间接地杀伤和抑制肝癌细胞。

These results suggest that in the central nervous system of small mammals, AP waveforms mature rapidly in many types of neurons, thus, coordinated activities in neuronal circuits may occur. In early postnatal development, GABA receptor mainly mediates an excitation effect, depolarizes P6 MGBv neurons and dramatically increases the frequency of AP firing. Inhibition of GABA receptor matures slowly along with increment of postnatal days.

结论大鼠MGBv神经元的AP波形成熟较快,可在中枢神经系统的神经元环路中产生协调性的活动;在MGBv神经元发育早期,GABA主要作为兴奋性神经递质而发挥作用,它可使P6的MGBv神经元产生去极化,AP发放频率显著增加,随着出生后日龄的增加,GABA受体的抑制作用才逐渐发育成熟。

Our study includes four aspects. In the first aspect we study several important conditions of porcine oocytes maturation in vitro and oocytes cleavage after parthenogenetic activation and found mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15% PFF+0.57mMcysteine is a good culture condition .When the Cocs are cultured in it ,the maturation rate and oocytes cleavage rate are higher than those of foreign covered. Our result are (86.7±3.35)% and (86.3±4.16)% and the highest report of foreign is(85.7±4.1)%.In the second aspect we study the effect of different chemical activations on development of porcine parthennogenetic embryo and found two best activation method. The first one is that putting the maturation MII oocytes in the 20μmol/L ionomycin for 30 minutes and then putting them in the NCSU-23 condition containing 5μg/mICB and 5mM/L6-DMAP for 3.5 hours, the oocytes cleavage rate and morulae/blastocysts development rate are (76.7±7.6)% and (37.1±6.4)%.The second one is that putting the maturation MII oocytes in the 200μM/L Thimerosal for 20 minutes and then putting them in the NCSU-23 condition containing 8mM DTT for 30 minutes

本研究分为4个部分,第一部分对影响猪卵母细胞体外成熟和孤雌激活后胚胎分裂的几个重要条件进行了比较研究,确立了一种较好的培养方法:与颗粒细胞共培养,找到了一种适合猪卵母细胞体外成熟的培养基:mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15%PFF+0.57mM半胱氨酸,成熟率和分裂率分别为(86.7±3.35)%和(86.3±4.16)%,国外报道的最高成熟率为(85.7±4.1)%;第二部分对猪卵母细胞孤雌激活的化学方法进行了研究,确立了化学激活猪卵母细胞的两种最佳方法:1将成熟的去卵丘颗粒细胞的MII期卵母细胞用20μmol/Lionomycin作用30min,再将卵母细胞培养于含5μg/mlCB和5mM/L 6-DMAP(6-二甲基氨基嘌呤)的NCSU-23培养液中,卵裂率和桑囊胚发育率达到(76.7±7.6)%和(37.1±6.4)%2将成熟的去卵丘颗粒细胞的MII期卵母细胞在200μM/L的Thimerosal中处理20min,再与8mM的DTT共孵育30min,卵裂率和桑/囊胚形成率为(81.0±2.8)%和(39.6±2.7)%;第三部分对孤雌激活胚胎的培养条件进行了研究,确立了一种最佳的胚胎培养条件:在SOF简单培养基中添加颗粒细胞进行前3天的培养,然后转入添加胎牛血清的NCSU—23培养基并和输卵管上皮细胞进行后期的培养,其桑椹胚和囊胚的发育率为(59.5±3.2)%;第四部分研究了IGF-I

The meiosis of oocytes differ from mitoses of general cells. Especially in most mammalian species, oocytes are formed during fetal life, but they are arrested at the prophase stage of the first meiotic division until around the time of ovulation.

卵母细胞的成熟分裂有别于一般细胞的有丝分裂,尤其是哺乳动物,在胎儿期或出生后不久,初级卵母细胞就陆续进入第一次成熟分裂前期阶段,但长时间的&休止&于此期,直到性成熟时在促性腺激素等因素的作用下,第一次成熟分裂才重新启动。

Japonicus.Nerve extraction substance of sea star and dithiothreitol has been reported to be effectively to induce oocyte maturation in sea cucumbers. In this study we tested their effects on oocyte maturation induction in sea cucumber A. japonicus. The results are as follows, oocyte of A. japonicus does not get mature spontaneously. Neither NES alone nor NES with follicle suspension can induce oocyte maturation. While DTT can prominently increase the percentages of germinal vesicle break down. When the concentration of DTT is between10-1 mol/L and 10-3 mol/L, the effect of maturation induction is more sufficient. When the concentration is about 10-2 mol/L, the percentage of GVBD can reach up to more than 90%.

分别利用在海参中有诱导作用的海星神经提取液和二硫苏糖醇对从成熟卵巢解剖出的刺参卵母细胞进行诱导成熟试验,结果显示,刺参的卵母细胞在海水中不会发生自发成熟,海星神经提取液以及神经提取液加滤泡悬浮液对卵母细胞没有诱导成熟的作用,而二硫苏糖醇处理可以诱导卵母细胞生发泡破裂,当DTT溶液的浓度处于10-1 mol/L到10-3 mol/L之间时,对刺参卵母细胞的促熟作用较为明显,在10-2 mol/L左右时,卵母细胞的诱导成熟率可以达到90%以上。

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