研究不同氨基酸模式对奶牛乳腺上皮细胞生长的影响

本试验以泌乳奶牛乳腺组织为原料,采用组织块培养技术,研究培养基内不同氨基酸模式对奶牛乳腺上皮细胞增殖的影响。结果表明,随着培养天数的增加,细胞数量逐渐增加,第10天各组细胞数量分别达到最高,乳腺上皮细胞生长曲线呈S形。不同氨基酸模式对乳腺细胞生长有显著影响,与氨基酸不平衡组相比,氨基酸平衡组、理想组更能促进细胞生长,各组间差异显著(P0.05)。     

Effect of Estradiol-17β on protein synthesis and degradation rates in fused bovine satellite cell cultures

Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Further, in vivo studies indicate that estradiol (E2) affects muscle protein synthesis and/or degradation, but in vitro results are inconsistent. We have examined the effects of E2 treatment on protein synthesis and degradation rates in fused bovine satellite cell (BSC) cultures. Additionally, to learn more about the mechanisms involved in E2-enhanced muscle growth, we have examined the effects of compounds that interfere with binding of E2 or insulin-like growth factor (IGF)-1 to their respective receptors on E2-induced alterations in protein synthesis and degradation rates in BSC cultures. Treatment of fused BSC cultures with E2 results in a concentration-dependent increase (P < 0.05) in protein synthesis rate and a decrease (P < 0.05) in protein degradation rate. The pure estrogen antagonist ICI 182 780 suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation in fused BSC cultures. The G-protein coupled receptor (GPR)-30 agonist G1 does not affect either synthesis or degradation rate, which establishes that GPR30 does not play a role in E2-induced alterations in protein synthesis or degradation. JB1, a competitive inhibitor of IGF-1 binding to the Type 1 insulin-like growth factor receptor (IGFR-1), suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation. In summary, our data show that E2 treatment directly alters both protein synthesis and degradation rates in fused BSC cultures via mechanisms involving both the classical estrogen receptor (ER) and IGFR-1.     

乙酸钠对油酸诱导的BRL-3A细胞脂肪变性的影响及机制分析

通过向油酸诱导的大鼠肝成纤维细胞(BRL-3A)脂肪变性模型中添加不同浓度(2、4、8 mmol·L^-1)的乙酸钠,探讨其对脂肪变性细胞模型脂代谢的调控机理及细胞损伤的修复作用。试验方法：1)用不同浓度的油酸(0、0.03、0.06、0.12、0.24、0.48) mmol·L^-1刺激BRL-3A细胞24 h后,分别检测细胞相对活力、总脂滴面积、三酰甘油(TG)含量、天门冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)活性,建立脂肪变性细胞模型;2)向BRL-3A细胞中添加不同浓度的乙酸钠,通过流式细胞术检测细胞凋亡率;3)用不同浓度的乙酸钠和0.12 mmol·L^-1油酸共同孵育BRL-3A细胞,试验分为4组,分别为油酸处理组、2 mmol·L^-1乙酸钠+油酸处理组、4 mmol·L^-1乙酸钠+油酸处理组和8 mmol·L^-1乙酸钠+油酸处理组,分别对细胞脂滴、TG含量、AST、ALT活性、AMPK信号通路蛋白以及脂代谢关键基因进行检测。结果显示:1)用0.12 mmol·L^-1油酸处理BRL-3A细胞24 h,成功建立BRL-3A细胞脂肪变性模型。2)不同浓度的乙酸钠对BRL-3A细胞凋亡率没有影响;3)4、8 mmol·L^-1乙酸钠处理脂肪变性细胞模型后,与油酸处理组相比,细胞总脂滴面积、每平方毫米脂滴数、TG含量、AST和ALT活性均显著(P<0.05)或极显著(P<0.01)下降,P-AMPK表达水平显著(P<0.05)或极显著(P<0.01)上升;脂合成代谢相关基因ACC、FAS以及SCD-1 mRNA表达水平均有一定程度下降;脂分解代谢相关基因CPT-1、CPT-2以及ACO mRNA表达水平均有一定程度上升。本研究表明,乙酸钠会通过AMPK通路激活脂分解代谢,减轻肝细胞脂质蓄积,并且对油酸诱导的BRL-3A细胞脂肪变性模型的损伤具有一定缓解作用。     

Advanced maternal age induces fetal growth restriction through decreased placental inflammatory cytokine expression and immune cell accumulation in mice

Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.     

复合双歧杆菌制剂对肉羊生长性能、养分表观消化率及血液生化指标的影响

为研究复合双歧杆菌制剂对肉羊生长性能、养分表观消化率及血液生化指标的影响,试验选择健康、体重相近的杜寒杂交肉羊60只随机分成4组,每组15个重复,每个重复1只,1组为对照组饲喂基础日粮,试验2、3、4组分别在基础日粮中添加2.5％、5.0％、10％的复合双歧杆菌制剂,预试验7 d,试验期56 d.结果表明:(1)试验3...     

PULSED WAVE-DOPPLER ULTRASONOGRAPHIC EVALUATION OF THE COMMON CAROTID ARTERY IN THE RESTING HORSE: PHYSIOLOGIC DATA

A pulsed wave-Doppler ultrasonographic evaluation of common carotid arterial blood flow was carried out on 63 healthy Italian Saddlebred horses. Vessel diameter and tracing morphology were evaluated were evaluated and blood flow parameters (systolic, diastolic and mean velocity, acceleration and deceleration of the systolic wave, cartid pulse volume) were calculated and correlated with class variables (sex, age and body weight). On the basis of the presence of an incisure in the ascending branch of the systolic curve, subjects were divided in two groups: one with a bifid systolic curve and the other with a monophasic aspect. Correlations between: 1) diameter of the vessel and body weight and 2) carotid pulse volume and flow velocity (systolic, diastolic and mean) were found. A greater systolic pulse volume was found in male subjects, in subjects with greater body weight and in those which had a monophasic systolic wave.     

柔嫩艾美球虫（Eimeria tenella）体外细胞培养

试验比较了玻璃珠层析法,ＤＥＡＥ纤维素层析法,Ｇ３漏斗过滤法３种提纯柔嫩艾美球虫（Ｅｉｍｅｒｉａｔｅｎｅｌｌａ）子孢子的方法,柱高８ｃｍ,直径２００μｍ的玻璃株柱用ｐＨ８．０的Ｒｉｎｇｅｒ’ｓ缓冲液作洗脱提纯子孢子,回收率为６４．１％,子孢子洗脱高峰期集中,且易于无菌操作,柱高５ｃｍ的ＤＥＡＥ纤维素柱用ｐＨ８．０的甘氨酸缓冲液作洗脱液,提纯子孢子回收率为６３．２％,但严格无菌操作困难,Ｇ３漏斗法易     

猪瘟免疫猪接种PRRSV Nsp2Δ1882-2241弱毒疫苗后免疫应答及T细胞表型变化特征

选择17头28日龄的CSFV和PRRSV抗体均为阴性的仔猪,于试验的第1天和第14天分别对其进行猪瘟耐热保护剂活疫苗（兔源）和高致病性猪繁殖与呼吸综合征Nsp2A1882—2241弱毒疫苗免疫。在免疫后的第28、42天采集外周血液,分析特异性抗体表达量和外周血T淋巴细胞表型的变化,评估猪瘟免疫对猪繁殖与呼吸综合征免疫的影响。结果显示,在CSF免疫后第28、42天,CSFV高抗组中的CD4^＋、CD4^＋／CD8^＋、CD4^＋CD8^＋和CD4-CD8数目均比CSFV低抗组高,CD3^＋和CD8^＋细胞数量比CSFV低抗组低;PRRS高抗组中,CD4CD8-细胞含量高于PRRS低抗组;在CSF免疫后第28天,CSFV抗体产生较高（阳性比率为73．33％）,PRRSV抗体产生较低（阳性比率仅为6．67％）。在CSF免疫后的第42天,CSF高抗组中PRRSV抗体阳性比率较CSF低抗组高8．33％。结果表明,CSFV特异性抗体产生高时能增加PRRSV特异性细胞免疫应答,增加CD4^＋细胞、CD4^＋CD8^＋细胞数量,提高机体免疫水平。CD3＋和CD4CD8-细胞应答作用值得重视。     

适应无血清培养的HEK293细胞系的培育

为培育适合腺病毒增殖的无血清培养细胞系,本研究采用培养液渐进替换的方法获得了一株适应无血清培养的HEK293细胞(293SF),经检测293SF细胞具有稳定的支持腺病毒增殖与表达外源蛋白的能力;平均病变时间为97 h,比HEK293细胞缩短12.9%;毒价达到107.48TCID50/mL,比HEK293细胞提高43.3%;并且稳定性良好,连续传16代后293SF细胞状态与易感性未发生变化。该细胞系为腺病毒的无血清培养奠定基础。