猪原始生殖嵴细胞(PGCs)建系因素的研究

从五指山猪(WSZP)近交系第8～13代培育群中，先后选用21头5～10月龄青年母猪，分别于授精后25～30d采集胎儿106个，进行原始生殖嵴(PGCs)细胞分离、培养等建系技术研究。以DMEM F10(1:1)为基础培养液，按添加或不添加生长因子，将培养液分为A、B、C3种，并以STO细胞作饲养层，在38℃、5.0％CO2和湿润的气相中进行培养建系。结果获得胚胎生殖嵴细胞(EG)细胞系6个细胞株，其中1个EG细胞株传至11代、2个传至5代、1个传至4代、2个传至3代冻存。并进行了AKP染色、体外分化、冷冻-解冻复苏和嵌合体制作等鉴定研究。研究发现：不同胚龄对EG细胞建系具有一定影响，不同培养液对EG细胞建系效果不同，STO细胞饲养层的质量是建株、传代、冷冻-解冻复苏的关键因素之一。EG细胞系的初步建立，为今后筛选进入种系的EG细胞系、实施体外基因操作提供了可能。     

金华地区牧草引种试验研究初报

对从国外和北京引进的13个牧草品种在金衢盆地进行了栽培及品比试验。结果表明：籽粒苋、苏丹草、俄罗斯饲料菜、甜高梁等鲜草产量高；紫花苜蓿、白三叶、鲁梅克斯、欧洲菊苣、籽粒苋等牧草粗蛋白含量较高，在20％以上；冬牧70黑麦草出苗后可在严寒的冬季生长，且产量高；特早熟二倍体和意大利四倍体黑麦草生育期长，可延长黑麦草利用期1个月左右。     

山羊孤雌胚胎的体外培养

系统地研究了培养液微滴的大小、培养液体系及血清浓度对山羊孤雌胚胎早期体外发育的影响。结果表明，培养液微滴以较大体积(200～500μL)的培养效果较好；在3种培养液中，添加10％的NGS对山羊孤雌胚胎的体外发育效果较好，囊胚率分别可达62．79％(81／129)、53．52％(38／71)、13．64％(12／88)；mCRlaa组囊胚发育率和囊胚细胞数显著低于SOFaa组和CRlaa组，SOFaa组优于CRlaa组，尽管SOFaa组和CRlaa组间无显著差异。在本试验条件下，以SOFaa培养液，山羊孤雌胚胎体外培养72h时加入10％的NGs，500μL微滴培养，其发育效果较好，囊胚率可达62．79％。     

山羊胎期肺发育的形态学研究

对3～22周龄山羊胎儿肺进行了肉眼、光镜和透射电镜观察，结果表明：1．7～22周龄山羊胎儿肺的外部形态与胎龄无关，肺的外部形态以左二右四叶者为多见．肺的叶间裂和右肺副裂常不完整，以浆膜、肺组织或混合性组织(肺组织及浆膜)融合；2．山羊胎儿肺的发育分为5个时期：胚胎期(3～5周)肺芽分支形成主支气管，主支气管长度不断增长并萌芽出叶支气管，均衬以假复层柱状上皮。腺状期(6～12周)以支气管树发育为主，小支气管衬以假复层和／或单层柱状上皮；终蕾呈腺状，上皮细胞由假复层柱状逐渐变为单层柱状，胞核向细胞顶端移行；终蕾上皮细胞游离面可见短小的微绒毛；线粒体、粗面内质网及核糖体随着胎龄增加而逐渐增多，它们均位于细胞顶部。小管期(13～14周)以呼吸部发育为主，原始肺泡开始形成，呼吸性细支气管衬以未分化的立方上皮；终蕾腺状结构逐渐消失，终蕾上皮细胞由高矮不等的单层柱状上皮逐渐演变为立方形的原始肺泡上皮；细胞游离面可见较多的微绒毛，胞质内线粒体、粗面内质网及核糖体较发达。囊状期(第15周)呼吸部发育显著，肺内细支气管及其末端呈现出“充气”状态；部分原始肺泡上皮细胞分化为扁平的肺泡Ⅰ型细胞和立方形的肺泡Ⅱ型细胞；Ⅱ型细胞内出现嗜锇小体。肺泡期(16～22周)以肺泡的形成和分化为主，更多的肺泡上皮分化为扁平的肺泡Ⅰ型细胞和立方形的肺泡Ⅱ型细胞。此期，毛细血管内皮与部分肺泡上皮贴近，可将肺泡上皮细胞区分为3种：Ⅰ型细胞，呈矮柱状或椭圆形，胞质中有较明显的核糖体、扩张内质网及变性线粒体；形成了由Ⅰ型细胞一基膜一内皮细胞组成的气血屏障。Ⅱ型细胞，胞质内含丰富的嗜锇板层小体和核糖体，内质网扩张呈大小不一的泡状，多泡体出现，线粒体膨大变性，细胞游离面可见少数微绒毛。Ⅲ型细胞，为未分化细胞，呈立方形，胞体较小，胞核相对较大，呈圆或椭圆形，胞质少，呈带状．电子密度低，细胞器少。     

耕作方式与长期定位施肥对雨养农田冬小麦产量的调控效应

为了明确西北黄土高原半湿润偏旱区耕作方式与长期定位施肥对冬小麦产量的调控效应,以设在半湿润偏旱区连续12年的耕作与肥料长期定位试验为平台,采用裂区设计,以传统耕作和免耕耕作为主处理,不施肥(CK)、单施无机氮肥(N)、单施无机磷肥(P)、单施有机肥(M)、无机氮磷肥配施(NP)、有机无机肥配施(NMP)为副处理,栽培制度为1年春玉米-3年冬小麦轮作,研究耕作及施肥措施对冬小麦产量的影响及其在生产年型间的变化关系。结果表明,不同年型及耕作方式均以有机无机配施冬小麦产量最高,有机肥单施高于化肥单施,磷肥单施高于氮肥单施。耕作方式间表现为传统耕作高于免耕耕作,年型间表现为丰水年>平水年>干旱年,耕作和施肥方式的增产效果以干旱年最好,平水年和丰水年差异不显著。有机无机配施与传统耕作结合优化了冬小麦冠层温度、叶绿素相对含量等生理指标,提高了光合速率、气孔导度及蒸腾速率,因而改善了有效穗、穗粒数和千粒重等产量性状而使冬小麦增产。在550 mm左右降水量的陇东旱塬雨养农业区,无论何种耕作方式及生产年型,长期采用有机无机或无机氮磷肥配施均表现出持续提高冬小麦产量的良好作用。因此,有机无机配施结合传统耕作是提高陇东半湿润偏旱区冬小麦产量的最佳耕作栽培模式。     

冷季放牧补饲对甘肃高山细毛后备母羊瘤胃代谢参数及瘤胃微生物数量的影响

旨在筛选出甘肃高山细毛羊后备母羊科学合理的补饲料,改变牧区由于营养供给不足导致甘肃高山细毛羊2.5岁才能配种的饲养方式。试验选择甘肃高山细毛羊(12月龄)48只,随机分为试验a(1号料)、b(2号料)、c(3号料)组和对照组(不补饲),进行90 d放牧补饲试验。结果表明,1)瘤胃pH试验a组和b组差异极显著(P<0.01)、c组显著(P<0.05)低于对照组,瘤胃氨态氮、蛋白氮、总氮浓度试验组均极显著高于对照组(P<0.01),总挥发性脂肪酸(VFA)试验a组和b组均极显著高于对照组(P<0.01),b组显著高于c组(P<0.05);乙酸和丙酸摩尔比a组极显著高于对照组(P<0.01),b和c组显著高于对照组(P<0.05);丁酸摩尔比a组极显著高于c组(P<0.01);乙/丙b组显著低于对照组(P<0.05)。2)瘤胃纤维降解菌和原虫数目试验组均极显著(P<0.01)或显著(P<0.05)高于对照组。综上所述,补饲均能正向影响甘肃高山细毛羊后备母羊瘤胃代谢参数及瘤胃微生物数量,以1号料补饲效果最优,2号料也能起到理想效果。     

低温胁迫对狗牙根生理及基因表达的影响

以耐寒性差异较大的杂交狗牙根品种运动百慕大、天堂419和普通狗牙根品种保定狗牙根为试验材料,分析了昼夜温度为适温(30 ℃/25 ℃)、亚适温(18 ℃/12 ℃)、冷害(8 ℃/4 ℃)和冻害(4 ℃/-4 ℃)4种温度处理下,低温对狗牙根生长速率和草坪质量、MDA含量、叶绿素荧光(F_v/F_m)、光合作用(P_n、G_s、C_i、T_r)、H₂O₂和O2·-含量、抗氧化酶活性(SOD、POD、CAT、APX)、抗氧化酶及耐寒相关基因(CBF1、COR、LEA)表达的影响。结果表明,随着温度的降低,狗牙根草坪质量、生长速率、F_v/F_m和光合作用显著下降,但在耐寒性强的运动百慕大中下降幅度较小;低温下叶片MDA和H₂O₂含量及O2·-产生速率显著升高,其中在耐寒性弱的保定狗牙根中升高程度更大;狗牙根叶片抗氧化酶活性及抗氧化酶基因的表达水平随着处理温度的下降而显著升高,其中在运动百慕大中更为明显;狗牙根CBF1、COR和LEA基因表达量随着温度的下降而显著升高,尤其是在耐寒狗牙根运动百慕大中更为显著。低温诱导的抗氧化酶和耐寒相关基因上调表达,增强了细胞内的抗氧化防御,有助于狗牙根植株维持较高的光合作用和细胞膜稳定性,延缓叶片的衰老,从而提高了狗牙根的抗寒能力。     

Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

Effects of oral administration of different dosages of carvacrol essential oils on intestinal barrier function in broilers

Essential oils are widely used in the pharmaceutical, food and cosmetic industries, and many plant essential oils have shown that they have positive effects on broilers nutrition. This experiment was conducted to study the effects of orally administered different dosages of carvacrol essential oils on intestinal barrier function in broiler chickens. A total of eighty 28‐day‐old (1.28 ± 0.15 kg) ROSS 308 broilers were randomly allocated to four groups of 20 replicates each, with one chicken per replicate per cage, and all were fed with the same diet. Four experimental groups were orally administered 0, 200, 300 or 400 μl carvacrol essential oils at 18:00 hr every day during the 2‐week experimental period. As a result of which, the gene expression of the occludin, claudin‐1, claudin‐5, ZO‐1 and ZO‐2 in intestinal mucosa of small intestine (p < 0.05) and the goblet cell content in small intestine epithelium (p < 0.05) were significantly increased; test subjects with 300 or 400 μl carvacrol essential oils reduced the microbial counts of Salmonella spp. and Escherichia coli in the intestines (p < 0.05); Essential oils administration also significantly increased activity of the sucrase (p < 0.05) and lactase (p < 0.05) in intestinal mucosa. In conclusion, the carvacrol essential oils have positive effects on growth performance and intestinal barriers function of broilers; those effects may be related to the dosage, as administration of 300 or 400 μl was more effective than that of 200 μl.     

用牦牛奶生产Scamorza奶酪技术研究

以新鲜牦牛乳为原料,研究了Scamorza奶酪的制作技术,采用四因素三水平正交试验的方法,研究了不同量的凝乳酶、氯化钙、食盐以及发酵温度对成品的口感、色泽质地的影响,最终确定了该产品生产的最佳配方及技术条件:凝乳酶0.003%、氯化钙0.01%、食盐22%,发酵温度为32℃.