高效毛细管电泳同时测定4种氟喹诺酮类抗菌药

建立同时测定4种氟喹诺酮类抗菌物(左氧氟沙星、培氟沙星、沙拉沙星和洛美沙星)的高效毛细管电泳(HPCE)法。考察了缓冲液离子浓度和pH、分离电压、运行温度等电泳参数,确立了最佳的电泳条件:缓冲液为10 mmol/L磷酸二氢钾-20 mmol/L硼砂(pH9.0,内含35mmol/L SDS),紫外检测波长278 nm,分离电压28 kV,运行温度25℃。结果表明,4种药物在8min内得到完全分离,分离度R为1.2以上。方法精密度(RSD)为0.22%~0.26%(迁移时间)和1.36%~2.39%(峰面积),回收率99.60%以上。该法快速、简便、准确。     

细胞壁降解相关酶以及在反刍动物应用中活性测定标准化的建议

针对当前国内外饲料酶制剂在反刍动物应用上结果不一致的现象,本文从酶制剂的角度,简要地分析了可能引起这些不一致的原因,同时就此总结了与细胞壁降解相关的主要的酶及其功能,并给出其活性测定标准化建议,为酶制剂的开发和应用提供参考。     

Teratoma Associated With Testicular Tissue in a Female-Like Horse With 64,XY (SRY-Positive) Disorder of Sex Development

A four-year-old female-like Quarter Horse was admitted for clinical evaluation because of masculinized-aggressive behavior and lack of estrous signs. On external inspection, a hypertrophied clitoris and prominent muscles were observed. On gynecological examination, apparently normal mammary glands, vulva, vagina, and cervix were noted. On the other hand, the uterus had no tone and was smaller than normal. The left gonad was very soft on palpation and the right gonad was mostly firm, irregular, and wider than the left gonad. On ultrasound examination, there were no signs of regular ovarian structure or follicular activity. Because of the different shapes and consistencies of the gonads and a suspicion of increased testosterone production, a bilateral gonadectomy was recommended. Blood was collected for testosterone levels quantification and for cytogenetic and molecular investigations. After removal, gonads were analyzed macroscopically and sections were sent for histopathological examination. A final diagnosis of benign adult teratoma associated with seminiferous tubules and Leydig cells was made. On cytogenetic and molecular analyses, a normal diploid number of 64 chromosomes and the presence of the XY sex chromosomes were seen in all cells, as well as the SRY gene. Testosterone levels were higher than normal before surgery and were reduced after gonads removal. In conclusion, the masculinized behavior was probably caused by increased testosterone levels produced by testicular tissue, in a female-like horse with 64,XY SRY-positive disorder of sex development, which was associated with a teratoma.     

Phenotypic plasticity induced using high ambient temperature during embryogenesis in domesticated zebrafish,Danio rerio

Ambient temperature during early stages of life has a substantial effect on physiological processes, eliciting phenotypic plasticity during zebrafish developmental stages. Zebrafish are known to possess a noteworthy ability to modify their phenotype in dependence of environmental factors. However, there is a poor understanding of the effects of temperature during embryogenesis, which influences the biological functions such as survival ability and masculinization in later developmental stages. Since the middle embryonic phase (pharyngula period) is genetically the most conserved stage in embryogenesis, it is very susceptible to embryonic lethality in developmental processes of vertebrates. Here, we tested the effect of transient perturbations (heat shock) during early development (5–24 hr post‐fertilization; hpf) at 35°C compared to control group at 28°C, on survival ability of zebrafish to study the embryonic and post‐embryonic mortality. We studied the variation of heat‐induced masculinization among and across the families in response to high temperature. Furthermore, morphometric traits of adult zebrafish at different developmental time points were measured in order to estimate the temperature × sex interaction effect. We found the highest embryonic mortality around the gastrula and segmentation periods in both experimental groups, with significantly lower survival ability in the temperature‐treated group (73.30% ± 0.58% vs. 70.19% ± 0.57%, respectively). A higher hatching success was observed in the control group (71.08% ± 0.61%) compared to the heat‐induced group (67.95% ± 0.60%). A distinct reduction in survival ability was also observed in both experimental groups during the first two weeks after hatching, followed by a reduced level of changes thereafter. We found sex ratio imbalances across all families, with 25.2% more males under temperature treatment. Our study on growth performance has shown a positive effect of increased temperature on growth plasticity, with a greater impact on female fish in response to high ambient temperature.     

Adiposity and adiponectin in dogs: investigation of causes of discrepant results between two studies

Although one study showed lower adiponectin concentrations in obese dogs, other recent studies indicate that adiponectin might not be decreased in obese dogs, raising the possibility that the physiology of adiponectin is different in dogs than in humans. The aim of this study was to investigate possible causes of the discrepancy between the two largest studies to date that assessed the association between adiposity and adiponectin concentration in dogs, including the validity of the assay, laboratory error, and the effects of breed, sex, and neuter status on the relationship between adiposity and adiponectin concentrations. Adiponectin concentrations measured with a previously validated adiponectin ELISA were compared with those estimated by Western blotting analysis of reduced and denatured plasma samples. The possibility of laboratory error and the effect of EDTA anticoagulant and aprotinin were tested. Adiponectin concentration was measured by ELISA in 20 lean dogs (10 male and 10 female, 5 neutered in each sex). There was close correlation between adiponectin concentrations measured by ELISA and those estimated by Western blotting analysis (r = 0.90; P < 0.001). There was no substantial effect of EDTA, aprotinin, or laboratory error on the results. There was confounding by neuter status of the relationship between adiposity and adiponectin concentrations, but adiponectin concentrations were not significantly lower in male than in female lean dogs (females, 36 mg/L; males, 26 mg/L; P > 0.20) and were not significantly lower in intact than in neutered lean male dogs (intact, 28 mg/L; neutered, 23 mg/L; P = 0.49). We conclude that the adiponectin ELISA previously validated for use in dogs appears to be suitable for determination of canine adiponectin concentrations and that testosterone does not appear to have a strong effect on plasma adiponectin concentrations in dogs. Obesity might decrease adiponectin concentrations in intact but not in neutered dogs.     

一种利用气相色谱快速检测瘤胃体外发酵液中三甲胺浓度的方法

本试验旨在建立一种利用气相色谱快速检测瘤胃体外发酵液中三甲胺浓度的方法.瘤胃体外发酵试验设置2个组,对照组(n=4)和试验组(n=4),试验组每个发酵瓶中添加0.0414 g三甲胺盐酸盐,发酵24 h.采用10 mol/L氢氧化钾溶液对发酵液样品进行预处理,使用WEL-PEG20M色谱柱和氢火焰离子检测器对三甲胺进行分...     

饲料添加剂L-赖氨酸硫酸盐中L-赖氨酸含量近红外速测方法研究

为了探讨利用近红外漫反射光谱技术(NIDRS)快速定量分析饲料添加剂L-赖氨酸硫酸盐中L-赖氨酸含量的可行性,本试验在全国范围内收集了具有代表性的L-赖氨酸硫酸盐添加剂76个,采用国家标准方法对样品中的L-赖氨酸含量进行化学赋值;用光栅型近红外光谱仪扫描L-赖氨酸硫酸盐样品,获取了不同物理状态下样品的近红外光谱图。依据L-赖氨酸含量将样品分为定标集和验证集,运用适当的光谱预处理方法,采用竞争性自适应重加权(CARS)算法结合偏最小二乘法(PLS)建立了L-赖氨酸硫酸盐的近红外定标分析模型,并将该模型与全波长模型进行了比较。结果表明:用烘干、60目粉碎后的样品结合CARS算法建立的定标模型最优,定标集校正决定系数(R2C)为0.954,校正集标准偏差(SEC)为0.510,交互验证标准偏差(SECV)为0.659;验证集预测决定系数(R2P)为0.952,预测标准偏差(SEP)为0.554,相对分析误差(RPD)值为3.83。由此可见,NIDRS定量分析L-赖氨酸硫酸盐具有一定可行性,对于丰富我国氨基酸盐及其他氨基酸制品的快速检测方法具有实际的应用意义。     

一种准确简便的东方白鹤性别分子鉴定方法

东方白鹤为国家Ⅰ级重点保护动物，在自然条件下繁殖率很低，进行迁地保护及再引入措施对遗传多样性保护具有举足轻重的作用，成功地进行性别鉴定是其笼养人工繁殖的前提。本研究对4只东方白鹤，运用CHD基因的一对引物2550F／27181K进行了准确简便的性别鉴定，并对其序列结合其他5种远缘鸟类相应序列进行分析，证实了此种方法的科学性和可信度，分析了性别鉴定机理。这种安全、方便、准确的性别分子鉴定方法将有助于东方白鹤迁地保护及再引入措施的实施。通过分析可以看出，此种方法未来存在着广阔的应用前景。     

利用分离的奶牛X精子冷冻精液生产体内性控胚胎试验

采用常规精液和分离的X精子冷冻精液对115头超数排卵的青年和经产荷斯坦母牛人工输精生产体内性控胚胎。结果表明,利用2支和3支性控精液给超排青年母牛输精,头均分别获得6.8枚±4.8枚和6.3枚±3.2枚可用胚胎,与常规精液的超排效果(7.2枚±5.2枚)无明显差异;利用2支和3支性控精液给超排经产母牛输精,头均可用胚胎分别为7.5枚±4.5枚和8.6枚±5.6枚,与常规精液的超排效果(6.1枚±3.2枚)相比,分别增加23%(P>0.05)和41%(P>0.05),无显著差异,但解冻后性控精子会明显影响经产母牛的受精率;对用性控精液输精生产的12枚抽样胚胎采用LAMP法进行性别鉴定并移植,结果获得了91.7%的雌性率和36.4%的受胎率。结果提示,X精子的分离过程会在一定程度上影响受精率,用分离的X精子冷冻精液对超排母牛进行人工授精生产体内性控胚胎是可行的。     

家蚕幼虫性腺表达序列标签分析(英文)

分别以5龄幼虫的精巢和卵巢构建了2个家蚕cDNA文库.利用表达序列标签方法分析参与家蚕性腺发育的基因,通过测序获得 16 737条ESTs(精巢 8 407条,卵巢 8 330条),拼接这些ESTs共得到 2 107个重叠群(contig)和 3 532个单拷贝(singlet).将这些转录物与GenBank非冗余蛋白质库比较,结果显示约34.3%的基因与数据库中蛋白质具有相似性.功能注释表明蛋白质合成相关基因、精巢特异基因等在文库中大量存在.基因本体(gene ontology)功能分类显示精巢与卵巢基因表达谱差异较大.研究结果为理解家蚕性腺发育提供了信息.