猪胸膜肺炎放线杆菌的分离与鉴定

从山东省泰安市采集的38份具有严重呼吸道症状的病死猪的肺脏、气管和扁桃体进行胸膜肺炎放线杆菌(APP)的分离鉴定,其中14株菌株表现出形态多样性、革兰氏染色阴性等特点,小鼠试验显示有较强的毒力,PCR电泳结果得到了650bp的预期目的片段。系统的生物学鉴定表明,这14株分离菌为胸膜肺炎放线杆菌(Actinobacillus Pleuropneaumoniae,APP)。对确诊的14株APP采用凝集试验进行血清型检测。结果从山东省泰安市分离到14株2个血清型的胸膜肺炎放线杆菌,其中,血清7型8株、血清5型6株,血清7型、5型为绝对优势血清型。     

Changes in Ovarian Follistatin Levels During the Oestrous Cycle in Sheep may Serve as an Intraovarian Regulator

The expression and concentration of follistatin and activin change during oestrous cycle suggesting their involvement in the regulation of follicular development. The aim of this study was to determine the level, source and potential role of follistatin in the sheep ovary. Follistatin in ovarian venous blood, measured by radioimmunoassay, remained at its low level from follicular phase (day ?1 and 0) to mid‐luteal phase (days 11–13) phase but were significantly elevated during the late luteal phase (days 14 and 15) when corpora lutea underwent regression. Western blot analyses of follicular fluid at day 15 of the cycle showed two strong bands at 42 and 45 kDa and weakly stained bands at 39 and 31 kDa. At day 0, these bands became weaker and the 39 kDa band became undetectable. However, there were no differences in follistatin concentrations between ovaries with and without functional corpus luteum (CL) during the whole luteal phase. In addition, although the ovaries of Booroola ewes normally contain more corpora lutea than those of normal merino ewes, follistatin concentrations in both jugular and ovarian venous blood were similar in Booroola and normal merino ewes. It is concluded that the secretion of follistatin from the ovary is not related to the formation of CL or high ovulation rate of Booroola ewes. The elevation in follistatin concentration in follicular fluid and ovarian blood during late luteal phase may indicate a dual role of follistatin in the luteolysis of existing CL and development of new follicle cohort.     

高效液相色谱-串联质谱法检测猪肝中苯并咪唑类药物及其代谢物的残留量

建立了猪肝中阿苯哒唑及其代谢物阿苯哒唑砜、阿苯哒唑亚砜、2-氨基阿苯哒唑砜,噻苯哒唑及其代谢物5-羟基噻苯哒唑,甲苯哒唑及其代谢物氨基甲苯哒唑、5-羟基甲苯哒唑,芬苯哒唑及其代谢物芬苯哒唑砜、等效物奥芬哒唑,氟苯哒唑及其代谢物2-氨基-氟苯哒唑共14种苯并咪唑类药物及其代谢物残留检测的高效液相色谱-串联质谱方法。样品用乙酸乙酯提取,MCX固相萃取柱净化。Waters Xterra C18色谱柱(2.1 mm×150 mm,5μm)分离,流动相A相为0.1%甲酸乙腈溶液;B相为0.1%甲酸水溶液,梯度洗脱,外标法定量。结果表明,14种苯并咪唑类药物及其代谢物标准溶液在10~1 000 ng/mL的浓度范围内呈现良好的线性关系,R2均大于0.99,方法检测限为5μg/kg,定量限为10μg/kg,14种苯并咪唑类药物及其代谢物在10~200μg/kg添加浓度范围内回收率均70%~120%之间。批内、批间相对标准偏差均小于20%。本方法灵敏、准确,满足食品安全检测法规的要求。     

Evaluation of in vitro chemosensitivity of vaccine-associated feline sarcoma cell lines to vincristine and paclitaxel

OBJECTIVE: To determine the in vitro sensitivity of 4 vaccine-associated feline sarcoma (VAFS) cell lines to the chemotherapeutic agents vincristine and paclitaxel. SAMPLE POPULATION: Cell lines derived from 4 VAFS specimens. PROCEDURES: Cell lines were cultured in vitro and individually exposed to various concentrations of vincristine and paclitaxel. Survival was estimated after 24 and 72 hours of exposure to each drug, and the drug concentrations that resulted in 50 and 90% reduction in number of viable cells (IC50 and IC90, respectively) were calculated. RESULTS: Both vincristine and paclitaxel had significant dose-dependent effects on the viability of the VAFS cell lines. After 72 hours of drug exposure, the IC50 and IC90 of vincristine for the 4 cell lines were between 0.005 to 0.039 microg/ml and 0.045 to 1.027 microg/ml, respectively. The IC50 and IC90 values for paclitaxel were between 0.037 to 0.092 microg/ml and 2.450 to 15.413 microg/ml, respectively. CONCLUSIONS: Results of pharmacokinetic studies on vincristine and paclitaxel in other species suggest that concentrations greater than the IC50 values may be possible for both drugs in feline patients as well. The drug concentrations at which viable cell numbers were reduced by 90% may also be attained in vivo for some cases, but detailed information is needed regarding the distribution, concentration, duration of availability, and toxicity of various drugs in cats. Carefully chosen combinations of antineoplastic agents need to be screened to identify treatment protocols that may be further evaluated clinically for the treatment of VAFS.     

苜蓿抗寒性及根蘖性状的表现与过氧化物酶同工酶关系的研究

测定了根蘖型和直根型苜蓿根、茎、芽的过氧化物酶同工酶的变化。结果表明,两类根系苜蓿同工酶谱带数目的变化均与季节温度变化相一致,但根蘖型苜蓿的数值明显大于直根型苜蓿,说明前者抗寒越冬力强于后者。变温处理后,４种抗寒性不同的苜蓿品种同工酶谱带数目及活性下降的顺序是：根蘖型苜蓿＜陇东苜蓿＜晋南苜蓿＜猎人河苜蓿。根蘖型苜蓿和直根型苜蓿根组织过氧化物酶谱明显不同,说明苜蓿根组织过氧化物酶同工酶的变化可能与控制根蘖性状的基因表达相关。     

甘草提取物对酒精性肝损伤的防治作用研究

以酒精性肝损伤小鼠为动物模型,探讨甘草提取物对酒精性肝损伤的防治作用.将64只昆明系小白鼠随机分为8组,即正常对照组、治疗组（低、中、高剂量）、预防组（低、中、高剂量）、模型组.模型组以56度白酒16 mL/kg灌胃,预防组和治疗组在酒精灌胃的基础上分别采用低（4.8 g/kg）、中（9.6 g/kg）、高（16 g/kg）不同剂量甘草提取物灌胃,连续14 d,取血液和肝脏样品,测定肝组织谷草转氨酶（GOT）、谷丙转氨酶（GPT）及血清乙醇脱氢酶（ADH）含量,同时采用H.E染色对肝组织进行组织学观察.血样测定结果表明,与模型组比较,低剂量预防组和治疗组GOT降低均不明显,中、高剂量预防组和治疗组GOT降低显著（P＜0.05,P＜0.01）;低剂量治疗组（P＜0.05）和其他给药组（P＜0.01）均能显著降低GPT含量;中、高剂量治疗组（P＜0.05）和低、中剂量预防组（P＜0.05）与高剂量预防组（P＜0.01）均能显著提升ADH含量;组织学观察表明,与模型组比较,中、高剂量治疗组及预防组能有效缓解和恢复肝损伤组织结构.提示甘草提取物对酒精引起的肝损伤具有一定的保护和治疗作用.     

细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用

为研究细胞信号转导和骨架在人源福氏志贺菌侵袭鸡肠上皮原代细胞中的作用,分别用酪氨酸蛋白激酶、酪氨酸蛋白磷酸酶、蛋白激酶C、Ca2+通道等信号转导抑制剂染料木黄酮、原钒酸钠、星状孢子素、硝苯地平和微丝蛋白聚合抑制剂细胞松弛素B处理鸡肠上皮原代细胞后,采用细胞免疫组织化学染色检测人源福氏志贺菌ZD02株对鸡肠上皮原代细胞侵袭率,用间接免疫荧光双重标记检测ZD02株侵袭鸡肠上皮原代细胞后F-肌动蛋白分布的变化。结果显示,染料木黄酮、硝苯地平和细胞松弛素B能显著抑制人源福氏志贺菌ZD02株内化,星状孢子素和原钒酸钠对ZD02株的内化无影响,人源福氏志贺菌侵袭鸡肠上皮原代细胞时F-肌动蛋白发生聚集。本研究表明人源福氏志贺菌ZD02株侵袭鸡肠上皮原代细胞过程中需要酪氨酸蛋白激酶、Ca2+通道活化以及细胞微丝重排。     

苜蓿自毒性的研究进展及其前景

从苜蓿自毒性的发现及概念入手 ,对苜蓿不同部分的自体毒素含量、不同土壤结构对自毒性的表现、不同的品种自体毒素含量的研究、自体毒素化合物对其它各类杂草或对作物的影响、苜蓿自毒性的可能物质基础、克服苜蓿自毒性的主要方法和途径等进行了论述。并展望未来可开展的研究领域以及我国开展相关研究的意义。     

家兔Zfx基因shRNA干扰载体构建及验证

研究主要是为了构建出针对家兔Zfx基因的shRNA重组载体及效果验证。根据家兔Zfx基因mRNA序列特征,设计出3条shRNA片段,再与线性化的pLL 3.7载体进行连接重组,最后经过qRT-PCR检测Zfx基因mRNA表达量筛选出有效重组表达载体并通过体内试验进行效果验证。结果表明:试验组shRNA-c和shRNA-f干扰效率分别为74%和67%,与空白对照组相比差异极显著(P<0.01);试验组shRNA-e干扰效果为17%,与空白对照组相比差异不显著(P>0.05)。用shRNA-c和shRNA-f进行体内试验,结果显示,两试验组雄性率分别为44.92%(P>0.05)和33.16%(P<0.01)。本试验成功构建针对家兔Zfx基因的重组载体,体内试验显示子一代出现性别偏移,说明Zfx基因对动物的性别决定有一定作用,关于导致体内验证出现反转的原因仍需进一步验证。     

中蜂对不同蜂花粉的取食偏好

以来自不同地区的10个蜂花粉,在云南及重庆两地的缺粉季节饲喂中蜂蜂群,探索中蜂对花粉取食的偏好。研究结果显示中蜂蜂群对来自不同地方不同种类的花粉有选择偏好性,对同种花粉选择差异性不显著,发现中蜂对所用的山东茶花粉的取食量最大。