UHPLC-Q/TOF MS结合HPLC-DAD定性和定量分析硫酸黏菌素可溶性粉中的未知添加物

定性和定量分析一批兽药硫酸黏菌素可溶性粉中的未知添加物。照《中国兽药典》2010年版一部对该批检品用微生物检定法进行含量测定时,发现该样品的抑菌圈为虚圈,用薄层色谱鉴别该样品,未显示与标准品溶液一致的主斑点,怀疑该样品中有处方外非法添加物。采用超高效液相色谱-四级杆-飞行时间质谱(UHPLC-Q/TOF MS)对该样品进行筛查,发现疑似添加物,并使用液相色谱-二极管阵列检测(HPLC-DAD)法进行了双重确证和含量测定。该样品中非法添加物确证为磺胺氯达嗪和甲氧苄啶,添加量分别为58.5 mg/g和13.4 mg/g。本研究通过建立筛查方法为监管部门提供技术支撑,通过分析非法添加物的可能原因为打击兽药处方外非法添加提供了思路。     

Based serum metabolomics analysis reveals simultaneous interconnecting changes during chicken embryonic development

Metabolic disorder is a major health problem and is associated with a number of metabolic diseases. Due to native hyperglycaemia and resistance to exogenous insulin, chickens as a model had used in the studies of adipose tissue biology, metabolism and obesity. But no detailed information is available about the comprehensive changes of serum metabolites at different stages of chicken embryonic development. This study employed LC/MS‐QTOF to determine the changes of major functional metabolites at incubation day 14 (E14d), 19 (E19d) and hatching day 1 (H1d), and the associated pathways of differential metabolites during chicken embryonic development were analysed using Metabolite Set Enrichment Analysis method. Results showed that 39 metabolites were significantly changed from E14d to E19d and 68 metabolites were significantly altered from E19d to H1d in chicken embryos. Protein synthesis was promoted by increasing the concentrations of L‐glutamine and threonine, and gonadal development was promoted through increasing oestrone content from E14d to E19d in chicken embryos, which indicated that serum glutamine, threonine and oestrone contents may be considered as the candidate indicators for assessment of early embryonic development. 2‐oxoglutaric acid mainly contributed to enhancing the citric cycle, and it plays an important role in improving the growth of chicken embryos at the late development; the decreasing of L‐glutamine, L‐isoleucine and L‐leucine contents from E19d to H1d in chicken embryonic development implied their possible functions as the feed additive during early posthatch period of broiler chickens to satisfy the growth. These results provided insights into understand the roles of serum metabolites at different developmental stages of chicken embryos, it also provides available information for chicken as a model to study metabolic disease or human obesity.     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

不同温度下圆叶决明降解过程红壤可溶性氮及氮水解酶动态变化研究

为了探明圆叶决明（Chamaecrista rotundifolia）降解过程果园红壤供氮水平的变化规律，本研究采用模拟培养试验，研究15℃和25℃培养果园红壤硝态氮、铵态氮、可溶性总氮、可溶性有机氮含量的变化及脲酶、蛋白酶、天冬酰胺酶活性的变化。结果表明，圆叶决明降解过程果园红壤4种可溶性氮含量均显著提高，且在培养140 d达到最大值，25℃培养的效果更佳。其中，可溶性总氮、可溶性有机氮和硝态氮含量随时间的动态变化可用三次曲线方程来拟合。圆叶决明降解过程还能显著提高脲酶、蛋白酶和天冬酰胺酶的活性，其中蛋白酶活性和脲酶活性的变化可用指数方程和三次曲线方程来拟合。本研究认为圆叶决明降解过程可提高果园红壤可溶性氮含量，因此可通过翻压圆叶决明提高土壤的供氮水平。     

南果梨周年干物质与氮磷钾积累动态

【目的】明确南果梨干物质积累特征和氮磷钾养分周年动态积累规律,为南果梨优化施肥量和施肥时期提供依据。【方法】以12年生南果梨树为试材,采用田间采样和树体分解方法,分别于萌芽后10 d(萌芽期)、 30 d(花期)、 65 d(幼果膨大期)、 100 d(果实膨大或新稍停止生长期)、 130 d(果实着色前)、 155 d(果实采收期)、 185 d(采收后)、 210 d(落叶前)8个生育期,选干周和树高一致的3株树,将树体连根从土壤中挖出,分出果实、 叶片、 枝条、 主干、 主根、 侧根、 须根,各器官单独称重,并取200 g左右的鲜样按清水、 洗涤剂、 清水、 1%盐酸、 3次去离子水冲洗、 杀青、 烘干后,电磨粉碎过0.15 mm筛,测定样品中氮、 磷、 钾含量。【结果】1)南果梨周年生育期内,树体干物质当年净积累量为18.4 kg/plant,干物质累积速率出现两次累积高峰,分别是幼果膨大期(0.15 kg/d)和采收期(0.11 kg/d)。2)南果梨树体总氮周年积累量为154.0~301.0 g,新生器官为0~116.2 g,果实膨大期达到最高;多年生器官氮积累量为154.0~194.8 g,落叶前达到最高。3)南果梨树体总磷周年积累量为17.1~37.2 g,果实着色前最高。其中新生器官为13.7 g,果实采收期最高;多年生器官为17.1~24.9 g,果实转色期最高。4)南果梨树体总钾周年积累量为27.9~174 g。新生器官钾为97.3 g,采收期最高;多年生器官钾为27.6~76.6 g,落叶前最高。5)产量大约为17 t/hm2的12年生南果梨从萌芽到落叶前树体当年氮磷钾的单株净累积量分别为146.2、 20.1、 146.1 g,折合1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。【结论】南果梨周年干物质单株积累总量为41.4 kg,当年净积累量为19.7 kg。南果梨干物质积累主要集中在花期到果实膨大期和果实转色到落叶前,分别占47.3% 和47.5%。南果梨从萌芽到落叶前氮、 磷、 钾的单株净累积量分别为146.2、 20.1、 146.1 g,每1000 kg果实经济产量需吸收氮(N)、 磷(P)、 钾(K)5.4、 0.7、 5.4 kg。从开花到果实膨大期和从果实着色到采收后30天对氮吸收分别占总氮累积量的39.0%和49.0%,而磷、 钾的累积从萌芽到开花较快,到果实膨大期磷的累积达67.4%,钾的累积达65.1%,果实膨大期是干物质和氮磷钾积累的关键时期。     

农五师葡萄钾素动态变化及调节措施研究

通过叶分析研究了不同葡萄品种叶柄全钾5～9月含量动态变化规律.结果表明:葡萄叶柄钾含量在生育期内整体呈波浪式下降趋势,前期下降迅速,中期平缓,进入浆果成熟期后又趋于下降；强势品种在整个生长季节内对钾的吸收大于弱势品种；早熟品种叶柄内钾含量变化趋势比晚熟品种提前.根据新疆农五师葡萄园区土壤有效钾含量分布现状,建议生产中应根据葡萄钾元素动态变化特点,不同团场应结合当地土壤有效钾含量相应补充.     

植物TIR1/AFBs基因家族研究进展

TIR1/AFBs基因家族是一种存在于细胞核中的生长素受体,属于F-box蛋白基因中的一个小亚族。它们通过与相关生长素相结合活化转录因子来促进基因的表达,从而进行调控,是生长素信号转导过程中的关键部分。为了深入研究生长素信号转导机制,从TIR1/AFBs基因家族的发现与结构,家族成员表达模式的差异及对植物生长发育方面的调节等方面概括介绍了TIR1/AFBs基因家族的分子调控机制,总结了TIR1/AFBs基因的功能。最后探讨了TIR1/AFBs基因的研究方向。     

1株鸭源非O1/O139群霍乱弧菌的分离鉴定与致病性分析

为探明引起贵州省某鸭场雏鸭发病的病原及其致病性和耐药情况，本研究对该鸭场送的疑似细菌感染病鸭进行剖检，取鼻黏膜、心脏和肝脏等组织器官接种于培养基中进行细菌分离鉴定，通过对分离菌进行药敏试验、动物回归试验和毒力基因检测研究其耐药情况和致病性。结果显示，分离菌在血琼脂培养基上生长16 h后呈现为边缘整齐、有光泽的乳白色菌落，伴有β-溶血现象，经革兰氏染色后在生物显微镜下呈两端钝圆、弧状、排列无规则的革兰氏阴性短小杆菌，与霍乱弧菌相符；16S rDNA基因序列同源性及系统进化树显示，该分离菌与霍乱弧菌同源性高达99.6%~99.7%聚为一支；药敏试验结果显示，分离菌对大部分药物都表现为耐药，其中对氨苄西林、克林霉素、复方新诺明、苯唑西林和克林霉素等抗菌药耐药性较强，对头孢哌酮和头孢曲松敏感；动物回归试验显示，分离菌可导致试验组雏鸭5 d内全部发病死亡，表明该分离菌对雏鸭具有较强的致病性；毒力基因PCR检测结果显示，检测的霍乱弧菌相关毒力基因hlyA、ompW和chxA为阳性，而检测的O1群rfb、O139群rfb、tcpA及ctxA基因为阴性，表明本次分离的霍乱弧菌携带有致病基因，但不属于O1和O139血清群。结果表明，该鸭场雏鸭发病的疫情病原为非O1/O139血清群霍乱弧菌，该菌致病性强且对多种抗菌药物耐药。本试验结果为贵州省鸭霍乱弧菌病的防控提供了参考依据。     

Histological and proteomics analysis of apple defense responses to the development of Colletotrichum gloeosporioides on leaves

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. is an important fungal pathogen known to cause glomerella leaf spot (GLS). The objective of this study was to analyze the infection of C. gloeosporioides on leaves of apple cv. Fuji (resistant) and cv. Gala (susceptible) and apply proteomics techniques to study the apple defense responses 48 h after inoculation (h.a.i.). On both of cultivars, C. gloeosporioides started to germinate at 3 h.a.i. on adaxial surface and produced appressoria adhering to epidermal cell juxtapositions. Histological analysis showed more stratified parenchyma in leaves of cv. Fuji than cv. Gala associated with differences in the chemical composition of cell walls. Total and unique proteins expressed by cvs. Fuji and Gala at 3, 12, 24 and 48 h.a.i. were detected by comparative proteomes analysis. A total of 42 unique proteins expressed at 24 and 48 h.a.i. were identified by MALDI/TOF mass spectrometry, and most of these proteins were identified as directly involved in defense responses.