表皮生长因子、巯基乙醇和亚硫磺酸对山羊体外胚胎生产的影响研究

采用抽吸法收集卵巢上的卵丘 -卵母细胞复合体 (COCs) ,微滴培养法于不同成熟培养液 (n =8)中培养 2 4～ 2 7小时后体外受精 ,然后移入不同胚胎培养液 (n =7)中培养 7天。应用胞核染色、胚胎移植等技术判定并统计成熟率、卵裂率和胚胎发育潜能。结果 :含 5 0ng/mlEGF(表皮生长因子 )组COCs成熟率 (75 4 5 %～ 75 8%)和卵裂率 (72 4 %～ 74.1%)均显著高于其它组 ,含 0 5mmol/LHTAU(亚硫磺酸 )组和或 10 0 μmol/LCYS(β -巯基乙醇 )成熟液组的≥ 8细胞率 (3 2 3 %～ 3 9 0 %)和桑胚率 (2 8 4 %～ 3 3 .8%)均显著高于其它组 (P <0 0 5 )。两只试管羊健康出生。结论 :EGF对COCs体外成熟和卵裂发育具有明显的促进作用 ;CYS对COCs体外成熟和受精卵裂无明显正作用 ,但成熟液含适量CYS有助于改善胚胎的体外发育 ,过量可能对细胞有毒 ;HTAU有益于早期胚胎的体外发育。成熟液中EGF和CYS的优选浓度分别为 5 0ng/ml和 10 0 μmol/L ;胚胎培养液中HTAU的优选浓度为 0 5mmol/L。优化体系所得胚胎具有正常的发育潜能。     

类胰岛素生长因子-I对鱼类氮代谢的影响

类胰岛素生长因子-I对鱼类的氮代谢起着重要的调控作用。类胰岛素生长因子-I可以通过影响蛋白质合成时转录模板DNA含量、转录效率和翻译水平而实现对鱼体蛋白质的调控。影响类胰岛素生长因子-I对鱼类氮代谢的调控因素包括水温、鱼的品种、不同的生长阶段、鱼体内的激素含量以及饵料的营养水平等。本文综述了鱼类类胰岛素生长因子-I的结构及其影响鱼类氮代谢不同作用方式等面的研究进展,并对类胰岛素生长因子-I以后在鱼类上可能的研究方向做了初步探讨。     

猪表皮生长因子真核表达产物的体外活性试验

为建立一种快速检测猪表皮生长因子(pEGF)活性的方法,用本实验室构建的pEGF酵母表达载体进行摇瓶培养,通过鉴定为所需要的目的蛋白,用pEGF为添加物培养Balbc/3T3细胞,以pEGF浓度为横坐标,吸光度为纵坐标建立曲线。结果表明,理想的接种细胞的个数为5 000个/孔,细胞计数试剂盒(CCK-8)孵育的最佳时间为1.5 h,pEGF和人表皮生长因子(hEGF)标准品,在1 ng/mL~100 ng/mL范围内对细胞的增值作用相当,表明pEGF和hEGF有相同的生物学活性。建立了用CCK-8快速测定pEGF活性的方法。     

重组猪表皮生长因子对早期断奶仔猪生长性能的影响

本试验旨在研究重组猪表皮生长因子(porcine epidermal growth factor,p EGF)对早期断奶仔猪生长性能的影响。选用21日龄断奶三元杂交(杜×长×大)健康仔猪160头,按体重相近、性别比例相同原则随机分成4组,即高档教槽料+500μg/kg p EGF组(GE组)、高档教槽料对照组(GD组)、中档教槽料+500μg/kg p EGF组(ZE组)、中档教槽料对照组(ZD组),每组4个重复,每个重复10头仔猪。试验期为10 d。结果表明:1)GE组平均日增重显著高于GD组(P<0.05),比GD组提高了6.29%;ZE组平均日增重极显著高于ZD组(P<0.01),比ZD组提高了17.08%;2)GE组平均日采食量与GD组无显著差异(P>0.05),ZE组平均日采食量显著高于ZD组(P<0.05),比ZD组提高了4.91%;3)GE组料重比显著低于GD组(P<0.05),比GD组降低了4.31%;ZE组料重比显著低于ZD组(P<0.05),比ZD组降低了10.37%;4)GE组腹泻率比GD组降低了57.14%;ZE组腹泻率比ZD组降低了38.46%。由此可见,饲粮添加p EGF提高了饲料转化率,降低了料重比和腹泻率,从而显著改善断奶仔猪生长性能,降低增重成本。     

Protein Localization of Epidermal Growth Factor in Sheep Ovaries and Improvement of Follicle Survival and Antrum Formation In Vitro

The aims of this study were to characterize EGF protein expression in ovine ovaries and to verify the effect of EGF on the in vitro development of isolated pre‐antral follicles. After collection, ovarian tissue was fixed for immunohistochemical analysis. Additional pairs of ovaries were collected, and secondary follicles were cultured for 18 days in α‐MEM⁺ (control) alone or supplemented with EGF (1, 10 or 50 ng/ml). The immunostaining for EGF was observed in oocytes from pre‐antral and antral follicles, in granulosa cells of primary and secondary follicles, as well as in cumulus and mural cells of antral follicles. After 18 days, the results showed that treatment with 50 ng/ml EGF significantly increased the percentage of morphologically normal follicles compared with the control group (α‐MEM⁺) and significantly reduced the precocious extrusion of oocytes and increased the percentage of antral follicles compared with the control and 1 ng/ml EGF. All the treatments induced a progressive and significant increase of the follicular diameter throughout the period of culture. However, there were no significant differences in follicular diameter or in the daily growth rate among treatments. In conclusion, this study demonstrated the presence of EGF in ovine ovaries. Moreover, 50 ng/ml EGF increased the percentage of normal follicles and improved antrum formation in isolated ovine follicles after 18 days of in vitro culture.     

In Vitro Loss of Tensile Strength and Elasticity of Five Absorbable Suture Materials in Sterile and Infected Canine Urine

The loss of breaking strength and elasticity of five absorbable suture materials (polydioxanone [PDS-II], polyglycolic acid [PGA], polyglactin 910 [PG-910], polyglyconate [GTMC], and chromic gut) after in vitro incubation in sterile, Escherichia coli- and Proteus mirab/tfs-inoculated canine urine was studied. Biomechanical testing, in a controlled environment, was performed during the 28-day study period. Polydioxanone and chromic gut retained greater than 90% of their original strengths after 28 days of incubation in sterile urine and 87% of original strengths in E. co//-inoculated urine. Polyglyconate retained 24% and 18% of original strength, respectively, after incubation in sterile and E. co/i-inoculated urine for 28 days. Polyglycolic acid and PG-910 retained less than 30% of original strength in sterile urine and only 7% in E. co/i-inoculated urine after 21 days of incubation. In P. m/rabi/is-inoculated urine, loss of tensile strength and elongation was significant for all suture materials. Polyglycolic acid and PG-910 lost all strength after 24 hours of incubation. Polydioxanone lost all strength after 7 days of incubation, whereas GTMC retained 19% at day 7. Chromic gut retained 78% at day 7 and 16% after 21 days of incubation, however, the absence of normal phagocytic destruction of chromic gut in this in vitro study may have artificially elevated these values. In sterile urine with chemically modified pH, loss of strength and elongation was greater in alkaline urine than in neutral or acidic urine for all types of suture materials.     

表皮生长因子在雌性哺乳动物生殖过程中的作用

表皮生长因子 (EGF)是表皮生长因子家族成员之一 ,在哺乳动物生殖过程中具有重要的调节作用。在哺乳动物卵巢内有 EGF的表达并调节卵巢的功能活动 ;EGF可促进哺乳动物卵泡发育 ,加速卵母细胞的成熟 ;EGF可促进卵母细胞受精和受精卵发育的能力 ,促进胚泡腔的形成 ;EGF在许多哺乳动物的子宫内均有表达 ,并参与胚胎着床过程的调节。 EGF可能以自分泌或旁分泌的形式参与卵泡发育、卵母细胞成熟、子宫内膜增殖、早期胚胎发育及胚胎着床等过程的调节。文章对 EGF在雌性哺乳动物卵泡发育、卵母细胞成熟、早期胚胎发育、囊胚腔扩展、胚胎着床等过程中的作用进行了概述     

表皮生长因子对成年水牛晶状体上皮细胞促增殖作用

为观察表皮生长因子 ( epidermalgrowth factor,EGF)对成年水牛晶状体上皮细胞增殖作用的影响。将不同浓度 EGF作用于体外培养的成年水牛晶状体上皮细胞 ,采用 MTT法测定细胞的增殖能力。结果 :EGF在浓度为 1ng/ m L和 1 0 ng/ m L作用的前 3 d促增殖作用逐渐加强 ,呈现时间依赖性 ,且在第 3 d达到最大促增殖效果。不同浓度的 EGF对晶状体上皮细胞的增殖作用不同 ,浓度在 1 0 ng/ m L作用 2 4h后即有明显的促增殖作用 ( P <0 .0 5) ,而作用72 h后最低的有效浓度为 1 ng/ m L,而且 2 50ng/ m L EGF有最大促增殖作用。结论 :EGF是诱导成年水牛晶状体上皮细胞增殖的重要因素。     

In Vitro Maturation of Dromedary (Camelus dromedarius) Oocytes: Effect of Different Protein Supplementations and Epidermal Growth Factor*

The present experiment was aimed to compare the effect of different protein supplementation sources, foetal calf serum (FCS), oestrous dromedary serum (EDS) and BSA, in experiment 1, and the effect of different concentrations of epidermal growth factor (EGF), in experiment 2, on in vitro nuclear maturation of the dromedary oocytes. Cumulus oocyte complexes (COCs) were harvested from the ovaries collected from a local slaughterhouse by aspirating the visible follicles in PBS supplemented with 5% FCS. Pooled COCs were randomly distributed to 4‐well culture plates containing 500 μl of the maturation medium and cultured at 38.5°C in an atmosphere of 5% CO₂ in air for 32–36 h. The basic maturation medium consisted of TCM‐199 supplemented with 0.1 mg/ml L‐glutamine, 0.8 mg/ml sodium bicarbonate, 0.25 mg/ml pyruvate, 50 μg/ml gentamicin, 10 μg/ml bFSH, 10 μg/ml bLH and 1 μg/ml estradiol. In experiment 1, this medium was supplemented with 10% FCS, 10% EDS or 0.4% BSA, whereas in experiment 2, it was supplemented with 0.4% BSA and 0, 10, 20 or 50 ng/ml of EGF. The oocytes were fixed, stained with 1% aceto‐orcein stain and their nuclear status was evaluated. Oocytes were classified as germinal vesicle, diakinesis, metaphase‐I, anaphase‐I (A‐I), metaphase‐II (M‐II) and those with degenerated, fragmented, scattered, activated or without visible chromatin as others. There was no difference (p > 0.05) observed in the proportion of oocytes reaching M‐II stage between the media supplemented with FCS (71.5 ± 4.8), EDS (72.8 ± 2.9) and BSA (72.7 ± 6.2). In experiment 2, a higher proportion (p < 0.05) of oocytes reached M‐II stage when the medium was supplemented with 20 ng/ml of EGF (81.4 ± 3.2) when compared with the media supplemented with 10 ng/ml (66.9 ± 4.1) and control (67.2 ± 7.1) groups. It may be concluded that the maturation media for dromedary camel oocytes can be supplemented with any of the three protein sources, i.e. FCS, EDS and BSA without any significant differences on the maturation rates. Also, a supplementation of 20 ng/ml of EGF in the maturation medium seems to be optimal and improves the nuclear maturation of dromedary camel oocytes.     

表皮生长因子对水牛卵母细胞体外培养核质成熟的影响

为了探讨表皮生长因子（EGF）对水牛卵泡卵母细胞体外培养核质成熟的影响，在以TCM199为基础的成熟液中加入不同浓度的EGF（0、10、25、50、100 ng/ml），体外成熟培养24~26 h，观察第一极体（PB1）的排放；随后进行孤雌激活检测其分裂率、囊胚发育率、囊胚孵化率，并用Hoechst33342染色后计算囊胚的细胞数。结果发现添加EGF各组的卵母细胞第一极体排放率显著提高（P<0.05）；成熟液中添加25 ng/ml EGF时，卵裂率及囊胚发育率（分别为80.0%、44.8%）明显高于对照组（分别为69.3%、31.9%，P<0.05），但对囊胚的细胞数影响不大。EGF不仅促进水牛卵母细胞体外培养的核成熟，而且有利于卵母细胞的胞质成熟，其中EGF的最佳浓度为25 ng/ml。     

表皮生长因子对早期断奶仔猪生长性能及肠道健康的影响

仔猪早期断奶是规模化、集约化养猪生产中关键技术之一。早期断奶的应用可最大限度发挥母猪的繁殖性能及养殖场的设备利用率,同时还能减少病原体的垂直传播,从而提高养猪业的经济效益。但是,早期断奶可能会因为环境及饮食成分的改变引发断奶应激,影响仔猪的生产性能及肠道健康。国内外研究表明饲粮中添加表皮生长因子(EGF)可提高断奶仔猪生长性能,促进肠道发育,提高胃肠道消化酶活性,提高免疫力,降低腹泻率。本文就EGF对早期断奶仔猪生产性能及肠道健康的影响做一综述,为EGF在仔猪饲粮中的应用提供参考。     

促卵泡素和表皮生长因子对鸡胚精原细胞增殖的作用

利用精原细胞-体细胞体外无血清共培养模型研究了促卵泡素(FSH)和表皮生长因子(EGF)对鸡胚精原细胞增殖的作用。结果表明:单独的FSH(10～100ng/mL)或EGF(10～100ng/mL)可显著增加精原细胞的数目以及增殖细胞核抗原的表达。EGF(10ng/mL)联合FSH(10ng/mL)具有加性效应,但更高剂量的EGF(100ng/mL)则降低了FSH的促进作用。因此,FSH联合适量的EGF可促进精原细胞的增殖。     

表皮生长因子和胰岛素样生长因子对水牛卵母细胞胞质成熟的影响

以皮层颗粒（CG）单层分布于质膜下做为卵母细胞胞质成熟的标志,利用CG荧光染色法,研究了不同浓度表皮生长因子（EGF）和胰岛素样生长因子（IGF-1）及其组合对水牛卵母细胞胞质成熟的影响。结果发现：（1）添加不同浓度的EGF（10、20、30、50ng／mL）都可以提高胞质的成熟率,但是组间差异不显著（P〉0．05）;皮质颗粒的分布随着EGF浓度的升高逐步由中间分布向皮层分布转变;（2）在成熟液中添加IGF-130ng／mL时效果较好,能显著提高卵母细胞胞质的成熟率;皮质颗粒的分布随着IGF-1浓度的升高逐步由中间分布向皮层分布转变,在添加IGF一130ng／mL时,在皮层分布最好,随着IGF-1量的进一步增加,皮质颗粒又向中间分布转变;（3）添加20ng／mL EGF＋30ng／mL IGF-1组卵母细胞体外成熟率高于添加30ng／mL的IGF-1组,显著高于添加20ng／m LEGF组和对照组（P〈0.05）。由此表明,EGF和IGF-1对水牛卵母细胞体外成熟有协同作用。     

Effect of Anti-Basic Fibroblast Growth Factor (Anti-bFGF) on In Vitro Embryonic Development in Rat

In this study, we aimed at the in vitro effects of anti-fibroblast growth factor-2 (anti-FGF-2 or anti-bFGF) on embryo culture in rats. In vitro effects of anti-bFGF on total embryonic development were investigated in 40 rat embryos (which were divided into four groups) (obtained from five pregnant females) at 9.5 days of gestation that were cultured in whole rat serum (WRS), and in WRS+ 2.5, 5, and 10 μg/ml anti-bFGF. After 48 h of culturing, the embryos from each group were harvested to be analysed morphologically according to a morphological scoring system and biochemically to obtain the embryo protein content. The morphological score, embryo protein content, somite number and crown-rump length of embryos indicated that embryos cultured in WRS+ anti-bFGF had significant embryonic retardation. Mean morphological scores for the embryos grown in WRS, in the presence of 2.5, 5 and 10 μg anti-FGF-2 were 61.4 ± 1.64, 46.3 ± 8.42, 27 ± 2.58 and13.6 ± 0.96 respectively. These results suggest that bFGF is very important for normal embryonic development and rat anti-bFGF neutralizes bFGF effect.     

表皮生长因子和胰岛素样生长因子-Ⅰ对断奶仔猪免疫功能的影响

选取21日龄断奶长白仔猪37头,按表皮生长因子(EG F)、胰岛素样生长因子-Ⅰ(IG F-Ⅰ)、基础日粮、自然哺乳随机分为4组,每组3重复,每个重复3头。EG F和IG F-Ⅰ的剂量为17.86μg/d,研究其对早期断奶仔猪免疫功能的影响。结果表明:EG F组和IG F-Ⅰ组脾淋巴细胞转化率均极显著(P<0.01)高于基础日粮组,空肠后段肠黏膜SIgA含量极显著(P<0.01)高于基础日粮组而且EG F组脾淋巴细胞转化率高于IG F-Ⅰ组。EG F组小肠各段黏膜上皮间淋巴细胞数极显著(P<0.01)高于基础日粮组;IG F-Ⅰ组空肠和回肠黏膜上皮间淋巴细胞数极显著(P<0.01)高于基础日粮组;EG F组回肠黏膜上皮间杯状细胞数极显著(P<0.01)高于基础日粮组;IG F-Ⅰ组回肠黏膜上皮间杯状细胞数极显著(P<0.01)低于自然哺乳组。表皮生长因子和胰岛素样生长因子-Ⅰ能提高早期断奶仔猪的免疫功能。     

不同浓度表皮生长因子(EGF)对鸡胚盲肠上皮细胞生长的影响

为了研究不同浓度表皮生长因子(EGF)对鸡胚盲肠上皮细胞生长的影响,通过体外培养鸡胚盲肠上皮细胞,添加不同浓度表皮生长因子(EGF,0、10、20、30、40 ng/mL)对鸡胚盲肠上皮细胞生长的影响,并对体外培养的鸡胚盲肠上皮细胞进行了碱性磷酸酶鉴定,细胞形态学观察贴壁率,测定生长曲线。结果显示:加入20 ng/mL的EGF,对鸡胚盲肠上皮细胞增殖的促进作用是极显著的(P0.01)。表明EGF作为重要的细胞因子之一,参与调节并促进鸡胚盲肠上皮细胞体外生长。     

利用转基因家蚕在蚕丝中表达重组人表皮生长因子

人表皮生长因子(human epidermal growth factor,hEGF)具有刺激细胞增殖的强大能力,是伤口愈合的关键因子.利用转基因技术,在家蚕后部丝腺利用丝素轻链基因(fibroin light chain,FibL)启动子驱动FibL-hEGF融合蛋白的表达.借助piggyBac转座子将FibL-hEGF融合基因导入到家蚕基因组,利用荧光检测总共获得8个转基因阳性卵圈,转基因阳性率为36.36％.PCR和反向PCR检测证明外源融合基因FibL-hEGF已被成功整合到家蚕基因组中,且随机插入到家蚕染色体的5个位点.Western blotting检测表明FibL-hEGF融合蛋白在家蚕后部丝腺中表达且成功分泌到蚕茧中,进一步证实了转基因家蚕的产生.本研究中利用piggyBac转座子介导的转基因技术成功获得了转基因家蚕并得到重组hEGF蚕丝,该蚕丝可用于制造伤口敷料.