人HIF-1α基因重组减毒沙门氏菌载体的构建及稳定性检测

利用RT-PCR方法从低氧预处理的A549细胞中获得HIF-1α基因片段,将其定向插入pIRES-SEQ质粒NheⅠ、MLuⅠ2个酶切位点之间,得到真核表达载体pIRES-HIF-1α;采用电转化方法将其电转化入减毒沙门氏菌Ty21α中,获得重组减毒沙门氏菌菌株TPH;转染A549细胞,RT-PCR法检测HIF-1α基因表达,ELISA方法检测HIF-1α蛋白表达;TPH菌株在氨苄青霉素(+)和氨苄青霉素(-)LB培养板上分别传代40代,每10代提取质粒进行PCR及酶切鉴定.结果表明:试验成功构建了携带HIF-1α基因的重组减毒沙门氏菌TPH,其可在体外细胞中稳定表达HIF-1α基因及蛋白,TPH可稳定遗传.     

高剂量辐照对鸡肉脯主要营养成分的影响

为研究高剂量辐照灭菌处理对宠物食品主要营养成分的影响,以鸡肉脯为材料,研究了4 kGy、6 kGy、8 kGy、10 kGy、15 kGy、20 kGy、50 kGy、100 kGy和200 kGy的60Coγ射线对其脂肪、蛋白质等营养成分含量的影响。结果表明:鸡肉脯经4～200 kGy辐照后,与对照相比,鸡肉脯中的脂肪含量差异比较明显,但与辐照剂量没有显著的线性相关性,且辐照后脂肪含量最高的(8 kGy)比对照中的含量高2.8%,最低的(20 kGy和100 kGy)比对照中的低5.6%;辐照后鸡肉脯中的蛋白质含量都高于对照,当辐照剂量达到8 kGy时蛋白质含量都显著高于对照中的含量;碳水化合物低于对照;粗纤维、17种氨基酸、微量元素(钙、铁、锰、镁、钠)等与对照无显著差异;锌、钾、磷的含量仅15 kGy、6 kGy和20 kGy处理与对照中的含量有显著差异;与对照相比,经50 kGy以下剂量辐照处理后,鸡肉脯的色泽、风味与滋味、口感无明显变化,可接受性程度好,当剂量达到100～200 kGy时颜色轻微变浅,呈棕色或棕褐色,但不影响品质,风味与滋味没有明显变化,可接受性程度良好。     

The effect of body condition on propofol requirement in dogs

ObjectiveTo determine if body condition score (BCS) influences the sedative effect of intramuscular (IM) premedication or the dose of intravenous (IV) propofol required to achieve endotracheal intubation in dogs.Study designProspective clinical study.AnimalsForty–six client–owned dogs undergoing general anaesthesia.MethodsDogs were allocated to groups according to their BCS (BCS, 1 [emaciated] to 9 [obese]): Normal–weight Group (NG, n = 25) if BCS 4–5 or Over–weight Group (OG, n = 21) if BCS over 6. Dogs were scored for sedation prior to IM injection of medetomidine (5 μg kg^?1) and butorphanol (0.2 mg kg^?1) and twenty minutes later anaesthesia was induced by a slow infusion of propofol at 1.5 mg kg^?1 minute^?1 until endotracheal intubation could be achieved. The total dose of propofol administered was recorded. Data were tested for normality then analyzed using Student t–tests, Mann–Whitney U tests, chi–square tests or linear regression as appropriate.ResultsMean ( ± SD) propofol requirement in NG was 2.24 ± 0.53 mg kg^?1 and in OG was 1.83 ± 0.36 mg kg^?1. The difference between the groups was statistically significant (p = 0.005). The degree of sedation was not different between the groups (p = 0.7). Post–induction apnoea occurred in 11 of 25 animals in the NG and three of 21 in OG (p = 0.052).ConclusionsOverweight dogs required a lower IV propofol dose per kg of total body mass to allow tracheal intubation than did normal body condition score animals suggesting that IV anaesthetic doses should be calculated according to lean body mass. The lower dose per kg of total body mass may have resulted in less post–induction apnoea in overweight/obese dogs. The effect of IM premedication was not significantly affected by the BCS.Clinical relevanceInduction of general anaesthesia with propofol in overweight dogs may be expected at lower doses than normal–weight animals.     

内蒙古阿拉善左旗红砂荒漠土壤盐分离子含量与其含水量相关关系研究

对内蒙古阿拉善左旗红砂荒漠土壤盐分离子含量和土壤含水量的分布规律进行了研究,并对两者之间做了相关和回归分析。结果表明,阿拉善左旗红砂荒漠土壤中盐分离子的含量,阳离子以Na+为主,阴离子以SO42-为主。在垂直分布上,Mg2+、Na+、Cl-和SO42-含量随着土层深度的增加而增加,而K+、CO32-和HCO3-含量随着土层深度的增加而减少。阿拉善左旗3个地区(乌力吉、长流水和孪井滩)的土壤含水量介于1.39%～8.01%之间,平均含水量为5.02%。3个地区中,红砂荒漠土壤所含盐分离子中的Mg2+、Cl-和SO42-均与其土壤含水量之间存在明显的线性关系。     

猪流感病毒分离株的特性及致病性

为了解猪流感病毒(swine influenza virus,SIV)分离株A/Sw/SH/1/2007(H1N2)的特性,对毒株的抗原位点、受体位点、潜在糖基化位点进行比较分析,并进行雏鸡、小鼠致病性试验.结果表明,A/Sw/SH/1/2007与北美经典株A/Sw/Tennes/1455/1977(H1N1) HA抗原位点最接近,HA1蛋白4个抗原位点中只有Ca位点有2个氨基酸改变,发生抗原变异,Sa、Cb抗原位点均只有1个氨基酸发生改变,不影响其抗原性;受体位点高度保守,只有183位发生改变;A/Sw/SH 1/2007有6个潜在的糖基化位点,在276位丢失了1个潜在的糖基化位点,但同时在274位出现了1个新的糖基化位点.NA蛋白抗原位点序列与广西分离株A/Sw Guangxi/13/2006(H1N2)同源性最高,除401位氨基酸发生G→R改变外,其余抗原位点均保守.耐药性分析显示,该病毒对金刚烷胺、扎那米韦药物均敏感.致病性试验结果表明,A/Sw/SH/1/2007对雏鸡无致病性,ICPI为0;肌注小鼠2周内死亡100％,滴鼻小鼠死亡80％,存活小鼠血凝效价为27,而与经典H1N1亚型SIV毒株只有较低的交叉凝集,HI为2 3.4～3.6.     

Acid–Base Parameters and Steroid Concentrations in Pre‐Ovulatory Follicles and Plasma of Lactating Dairy Cows with Spontaneous and Synchronized Oestrus or Follicular Cyst

The study aimed to compare the acid–base balance and steroid concentrations between follicular fluids (FF) of pre‐ovulatory follicles derived from a spontaneous oestrus (SO), synchronized or induced oestrus (IO) and follicular cysts (CYS) and between FF and blood in dairy cows. Forty‐two dairy cows were included in this study. The animals were allocated to three groups: SO (n = 23); IO (n = 11) using GnRH at day 0 and day 9 and PGF₂α at day 7; and animals with CYS (n = 10). The follicular fluids (FF) were aspirated from the cyst/pre‐ovulatory follicles (? ≥ 15 mm) after SO and after second GnRH dose in IO by transvaginal ultrasound‐guided ovum pick‐up technique. Blood samples (BL) were collected in heparinized vacutainer tubes. The oxygen tension (pO2) in FF of IO was higher (p < 0.05) than in SO and CYS groups. There were negative correlations (p < 0.001, r = ?0.89) between FF and blood pO2. The carbon dioxide tension (pCO2) and lactate level in FF of CYS group were higher (p < 0.05) than in SO and IO groups. There were negative correlations (p < 0.01, r = ?0.73) between blood and FF pO2. Oestradiol‐17β concentration in pre‐ovulatory follicles and plasma of the SO group was higher (p < 0.001) than in IO and CYS groups. Progesterone concentration in pre‐ovulatory follicles and plasma of the SO and IO groups was lower (p < 0.01) than in CYS group. Plasma androstenedione concentration in SO and IO groups was higher (p < 0.05) than in CYS group. In conclusion, acid–base parameters, E2 and P4 levels in the follicular fluid of both IO and CYS groups were deviated greatly from the physiological level (disturbances of intrafollicular/intracystic environment), which may affect the quality of both the oocyte and the granulosa cells.     

吸收剂量、贮藏时间及贮藏温度对辐照牛肉中2-十二烷基环丁酮的影响

分析了辐照牛肉中辐解产物2-十二烷基环丁酮在不同的吸收剂量(0.5~10.0kGy)、贮藏时间(0-40d)及贮藏温度(-20℃~4℃)下的变化规律。采用硅胶层析柱-气相色谱/质谱联用技术萃取分离并检测。结果表明,2-十二烷基环丁酮的浓度与吸收剂量呈线性正相关,随着贮藏时间延长其含量减少,贮藏温度对其没有显著影响。利用2-十二烷基环丁酮可鉴定牛肉是否经过辐照,在已知贮藏温度和时间的条件下,可评估辐照牛肉的大约吸收剂量范围。     

槟榔江水牛OXCT1基因的分子特征及组织表达谱分析

本研究对槟榔江水牛3-酮酸辅酶A转移酶1（3-oxoacid CoA-transferase 1,OXCT1）基因的完整CDS区进行了PCR扩增,并对其功能生物信息学和多组织差异表达进行了分析。以普通牛OXCT1基因序列（GenBank登录号：XM_006076397）为参考序列,采用Primer Premier 5.0软件设计引物序列,以提取的槟榔江水牛基因组DNA为模板,PCR扩增获得槟榔江水牛OXCT1基因mRNA序列,扩增产物经测序后利用ORF Finder软件进行开放阅读框识别,获得水牛OXCT1基因CDS区全序列,并对其进行生物信息学分析。结果显示,槟榔江水牛OXCT1基因CDS序列全长1 563 bp,编码520个氨基酸,蛋白分子式为C₂₅₀₉H₄₀₄₁N₆₈₇O₇₄₆S₂₃,分子质量为56.50 ku,理论等电点（pI）为8.69,不稳定系数为27.49,平均疏水性（GRAVY）为-0.097,该蛋白属弱亲水性蛋白。OXCT1蛋白无信号肽和跨膜结构,属于线粒体膜蛋白;包含pcaJ_scoB_fam和AtoD 2个保守结构域,且具有5个功能活性位点。槟榔江水牛与普通牛、藏羚羊等5个物种的OXCT1氨基酸序列同源性≥ 97%。对槟榔江水牛13个组织进行表达谱分析表明,在泌乳期,OXCT1基因在乳腺中表达量最高,在肾脏、垂体、脂肪、大脑、皮肤和肌肉中不表达;在非泌乳期,OXCT1基因在除脂肪以外的其他12个组织中都有表达。OXCT1基因在槟榔江水牛泌乳期乳腺组织中表达量最高,推测其可能参与了水牛的乳脂合成调控事件。本研究结果可为深入了解乳脂合成代谢及其调控机制提供依据。