三氧化二砷对鸡马立克氏病肿瘤细胞凋亡的影响

研究了三氧化二砷（As2O3）对诱导鸡马立克氏病（MD）肿瘤细胞株MDCC—MSB1细胞凋亡相关基因Fas、Caspase-3 mRNA表达及Caspase-3活性的影响，进而探讨了AS2O3诱导MD肿瘤细胞株凋亡的机制。以体外培养MDCC-MSB1为研究对象，经终浓度分别为0、2、4和8μmol／L的As2O3作用48h后，采用荧光显微镜观察细胞的形态学变化，DNA Ladder法检测细胞凋亡情况，RT-PCR方法检测Fas、Caspase-3 mRNA的表达水平，试剂盒比色法检测Caspase-3活性的变化。结果表明，AS2O3能引起典型的凋亡形态学变化，并出现典型的DNA Ladder，同时Fas、Caspase-3 mRNA的表达水平和Caspase-3活性均增加，组间差异均极显著（P〈0．01），呈现剂量依赖性。证实As2O3诱导鸡MD肿瘤细胞株MDCC-MSB1细胞凋亡是通过增加Fas、Caspase-3 mRNA的表达和提高Caspase-3活性来实现的。     

钙稳态失衡在氯化镧诱导MDCC-MSB1细胞凋亡中的作用

为探讨钙稳态失衡在LsCl3诱导MDCC-MSB1细胞凋亡中的作用,MDCC-MSB1细胞常规培养于RPMI1640培养液中,加入终浓度为0.5,1,1.5,2,2.5,3,3.5和4 mmol·L-1的LaCl3,继续培养24 h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca2+]i的变化.结果表明,在LaCl3浓度为0.5～4 mmol·L-1时,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca2+]i呈升高趋势,并呈剂量-效应关系.这表明LaCl3能抑制MDCC-MSB1细胞的增殖,并可能通过改变[Ca2+]i而诱导其发生凋亡.     

钙稳态失衡在氯化镧诱导MDCC-MSB1细胞凋亡中的作用

为研究钙稳态失衡在LaCl3诱导MDCC-MSB1细胞凋亡中的作用,将MDCC-MSB1细胞常规培养于RPMI-1640培养液中,加入终浓度为0.5、1.0、1.5、2.0、2.5、3.0、3.5和4.0mmol/L的LaCl3,继续培养24h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca2+]i的变化.在LaCl3浓度为0.5～4.0 mmol/L范围内,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca2+]i呈升高趋势,并呈剂量-效应关系.结果表明,LaCl3能抑制MDCC-MSB1细胞的增殖,并可能通过改变[Ca2+]i而诱导其发生凋亡.     

莲草多糖和黄酮对MDCC-MSB1细胞凋亡及其Bcl-2mRNA表达的影响

为了探讨莲草多糖和黄酮对诱导鸡MD肿瘤细胞凋亡的作用效果及其可能的作用机制,试验以鸡MDCC-MSB1细胞系为研究对象,通过观察细胞形态变化,分析细胞DNA片段化及半定量RT-PCR法检测莲草多糖和黄酮在安全浓度范围内对鸡MD肿瘤细胞凋亡及Bcl-2 mRNA表达的影响。结果表明,莲草多糖和黄酮能诱导MDCC-MSB1细胞凋亡,并降低MDCC-MSB1细胞内Bcl-2 mRNA的表达水平,致使细胞抗凋亡能力下降,进而发挥其药物作用。     

钙稳态失衡在氯化镧诱粕CC-MSB1细胞凋亡中的作用

为研究钙稳态失衡在LaCl3诱导MDCC-MSB1细胞凋亡中的作用,将MDCC-MSB1细胞常规培养于RPMI-1640培养液中,加入终浓度为0．5、1．0、1．5、2．0、2．5、3．0、3．5和4．0mmol／L的LaCl3,继续培养24h后,应用MTT法检测细胞增殖抑制率,DNA Ladder法和TUNEL法检测细胞凋亡,以Fura-2为荧光探针检测细胞内[Ca^2＋]i的变化。在LaCl3浓度为0．5-4．0mmol/L范围内,细胞的增殖抑制率增加,细胞凋亡数量和细胞内[Ca^2＋]i呈升高趋势,并呈剂量一效应关系。结l果表明,LaCl3能抑制MDCC—MSB1细胞的增殖,并可能通过改变[Ca^2＋]i而诱导其发生凋亡。     

钙离子超载在镉致大鼠肝细胞凋亡中的作用

拟探讨Ca2+超载在镉诱导大鼠肝细胞凋亡中的作用。采用两步灌流法获得大鼠肝细胞,经过24 h培养,用醋酸镉、醋酸镉与Bapta-AM共同处理肝细胞。用MTT法检测细胞存活率,倒置显微镜和荧光显微镜观察细胞形态和凋亡,流式细胞仪检测细胞内Ca2+浓度、ROS生成和ΔΨm变化。结果表明,随着镉剂量的增加和作用时间的延长,细胞变形,坏死细胞和凋亡细胞增多,镉可极显著提高细胞内Ca2+浓度和ROS生成量(P<0.01),极显著降低线粒体膜电位(P<0.01),Bapta-AM可显著(P<0.05)或极显著(P<0.01)降低镉的毒性。研究结果提示Ca2+超载在镉致肝细胞凋亡中发挥重要作用。     

钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用

旨在探讨钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用。以体外培养鸡胚脑神经元为研究对象,在含终浓度为0、1.5、22、.5 mmol.L-1MnCl2的DMEM培养液中培养24 h,应用流式细胞仪检测磷脂酰丝氨酸(PS)和线粒体膜电位(ΔΨm),琼脂糖凝胶电泳法(DNA Ladder)检测细胞染色质的断裂,以Fura-3/AM为探针检测细胞内游离钙离子浓度([Ca2+]i),用实时定量PCR法检测细胞内CaMmRNA的表达水平。结果表明,随着MnCl2浓度的升高,ΔΨm呈降低趋势,PS膜外翻增加,细胞染色质DNA发生损伤,产生明显的梯形条带,细胞内[Ca2+]i呈升高趋势,CaMmRNA的表达水平下降。结果提示,过量锰通过抑制CaM活性,引起神经元内[Ca2+]i升高,膜通透性发生改变,ΔΨm降低,从而导致鸡胚脑神经元发生凋亡。     

钙稳态失衡在镉诱导大鼠大脑皮质神经细胞凋亡中的作用

用不同浓度醋酸镉（0、5、10、20μmol／L）以及细胞内钙离子螯合剂BAPTA-AM（10μmol／L）单独或联合作用于大鼠大脑皮质神经细胞12h,利用流式细胞仪检测细胞凋亡率、细胞内[Ca2＋];、活性氧（ROS）水平以及线粒体膜电位（△ψm）。结果显示,与对照组相比,各镉染毒组细胞凋亡率、细胞内[Ca2＋]i和ROS水平显著或极显著升高（P〈0．05或P〈0．01）,△ψm显著或极显著降低（P〈0．05或P〈0O．01）。BAPTA-AM联合组与相应镉染毒组相比,细胞凋亡率、细胞内[Ca2＋]i和ROS水平降低,△ψm升高,部分组间差异显著或极显著（P〈0．05或P〈0．01）。结果表明,镉可诱导大脑皮质神经细胞凋亡,细胞内钙稳态失衡在镉诱导大脑皮质神经细胞凋亡中起着重要作用,凋亡机理可能与细胞内钙超栽引起ROS水平升高和△ψm下降有关。     

鸡TGFβ1对MDCC-MSB1细胞增殖、凋亡、迁移与侵袭的影响

旨在探讨鸡TGFβ1对MDCC-MSB1细胞增殖、凋亡、迁移与侵袭的影响。作者将构建的鸡TGFβ1过表达载体、干扰表达载体以及相应阴性对照转染MDCC-MSB1细胞,然后检测转染后各组细胞鸡TGFβ1的表达水平、细胞增殖能力、细胞周期与凋亡,细胞的迁移与侵袭能力。结果显示,与相应阴性对照相比,转染TGFβ1过表达质粒可显著上调MDCC-MSB1细胞的TGFβ1表达水平,显著抑制MDCC-MSB1细胞的增殖,且使G1期细胞增加、S和G2期细胞减少,同时增加细胞凋亡率,降低细胞的迁移与侵袭能力;转染TGFβ1干扰表达质粒可显著下调MDCC-MSB1细胞的TGFβ1表达水平,显著促进MDCC-MSB1细胞的增殖,G1期细胞减少、S和G2期细胞增加,同时降低细胞凋亡率,增加细胞的迁移与侵袭能力。结果表明,鸡TGFβ1可抑制MDCC-MSB1细胞增殖、迁移与侵袭,促进其凋亡。     

镉致大鼠大脑皮质神经细胞凋亡的机理及N-乙酰半胱氨酸的保护作用

对神经细胞用不同浓度(0、5、10、20 μmol/L)醋酸镉染毒和N-乙酰半胱氨酸(NAC)(100μmol/L)进行保护,通过流式细胞仪检测了细胞凋亡率、[Ca2+]i和活性氧(ROS)水平.结果表明,与对照组相比,各染毒组细胞凋亡率、细胞内[Ca2+]i和ROS水平明显升高,呈剂量-效应关系,除5 μmol/L组ROS水平外,均差异极显著(P<0.01).添加NAC组与相应染毒组比较,神经细胞凋亡率有降低趋势,但差异不显著(P>0.05);10、20μmol/L组细胞内[Ca2+]i和ROS水平显著降低(P<0.05或P<0.01).表明镉可诱导神经细胞凋亡,其机理可能与细胞内Ca2+和ROS水平升高有关,NAC对其有一定的保护作用.     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

中华人民共和国农业部公告 第267号

为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …     

鸡败血支原体垂直传播模式及其阻断方法

以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.