家蚕孤雌生殖相关蛋白PGL的鉴定与生物信息学分析

应用二维凝胶电泳(2DE)技术分离有性生殖蚕卵与孤雌生殖蚕卵的差异蛋白,通过基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF-TOFMS)分析获得大量小肽的序列特征。与已构建的家蚕cDNA文库及蛋白质数据库进行比对,确定一个在家蚕孤雌生殖过程中表达量显著上调的差异蛋白为磷脂酰肌醇蛋白聚糖(phosphatidylinositol glycan,PGL)。为了探讨PGL蛋白在家蚕孤雌生殖发生中的作用,采用cDNA末端快速扩增(RACE)技术扩增目的基因5′端片段,获得PGL蛋白基因的全长cDNA,生物信息学分析显示该基因cDNA全长1038bp,编码345个氨基酸,其编码蛋白理论分子质量为39.516kD,等电点为5.845,包含1个保守的GPI8结构域,信号肽概率为0.997,可能为分泌型蛋白,预测蛋白功能域包含酪蛋白激酶Ⅱ磷酸化、N糖基化、蛋白激酶C磷酸化等多个功能位点。依据PGL蛋白的理化性质、结构功能推测其有可能在不同程度上通过细胞信号转导途径参与了家蚕孤雌生殖发生过程并发挥重要作用。     

甘肃茵陈蒿超临界CO2萃取产物化学成分的差异性分析

采用超临界CO2萃取联合气相色谱-质谱法检测了茵陈蒿Artemisia capillaris萃取产物化学成分,并与水蒸气蒸馏法相比较。结果从超临界CO2萃取提取物成分中确定了76种化合物,从水蒸气蒸馏提取物成分中确定了48种化合物。其中长马鞭草烯酮含量分别为37.35%和26.68%,2种方法测得的成分有所差异。与传统水蒸气蒸馏法相比,采用超临界CO2法更能真实、全面地反映茵陈蒿的化学组分,实验为进一步研究利用茵陈蒿植物资源提供了科学依据。     

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.     

Simultaneous steroids measurement in dogs with hyperadrenocorticism using a column-switching liquid chromatography-tandem mass spectrometry method

We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2–16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.     

荷斯坦奶牛瘤胃微生物脲酶的分离与鉴定

本研究旨在从荷斯坦奶牛瘤胃中分离活性脲酶。从奶牛瘤胃中收集瘤胃内容物,通过离心和超声破碎的方法得到不含菌体细胞的瘤胃液(CFRF)和瘤胃菌体蛋白液(RCP),利用85%硫酸铵盐析,并透析后,用HiTrap Capto Q离子交换层析柱纯化脲酶。将纯化后的脲酶用活性PAGE分离,并利用改良的Fishbein染色法,确定脲酶条带位置,切胶,经胰蛋白酶消化后,用LC-MS/MS分析,并用SEQUEST软件在NCBI数据库搜索与质谱信号相匹配的肽段和蛋白质。结果表明从CFRF和RCP中分离出较高活性的脲酶,但经染色后只有RCP脲酶显示活性条带。经质谱分析,最终鉴定出了3种微生物来源脲酶,分别与嗜热链球菌(Streptococcus thermophilus)、唾液链球菌(Streptococcus salivarius)和嗜碱芽孢杆菌(Bacillus halodurans)相似。这说明绕过纯培养微生物,直接从瘤胃中分离脲酶,来研究其性质和来源是可行的,尤其适合对未培养微生物的研究。     

Clinical pharmacology of clemastine in healthy dogs

The pharmacokinetic properties of clemastine were investigated in six healthy dogs and compared with the effect of the drug recorded as inhibition of wheal formation induced by intradermal injections of histamine. Clemastine clearance was high (median: 2.1 L h(-1) kg(-1)) and the volume of distribution large (13.4 L kg(-1)). The half-life after intravenous administration was 3.8 h and the plasma protein binding level in vitro was 98%. After oral administration, the bioavailability was only 3%. Given intravenously, clemastine (0.1 mg kg(-1)) inhibited wheal formation completely for 7 h, whereas the effect after oral administration (0.5 mg kg(-1)) was minor. The data show that most dosage regimens suggested in the literature for the oral administration of clemastine to dogs are likely to give too low a systemic exposure of the drug to allow effective therapy.     

套袋对鸭梨果实香气及糖酸组分的影响

采用静态顶空和气相色谱-质谱联用技术、高效液相色谱技术,研究了生长期套袋对鸭梨果实挥发性成分、糖酸组分等风味物质的组成和含量的影响。结果表明:套袋导致鸭梨香气物质种类减少和相对百分含量降低。套袋鸭梨采收时测定的香气成分有3类18种,其中酯类15种、烯类2种、醇类1种;不套袋鸭梨采收时测定的香气成分有3类26种,其中酯类23种、烯类2种、醇类1种。鸭梨的主要香气成分是乙酸己酯、己酸乙酯、丁酸乙酯。套袋和不套袋鸭梨均检测到3种糖、4种有机酸组分,其中糖组分主要为果糖,其次为蔗糖、葡萄糖,有机酸组分主要为苹果酸,其次为乙酸、柠檬酸、草酸,但不同处理间含量存在差异。     

液相色谱—串联质谱法研究恩诺沙星在中华草龟体内的药物代谢动力学

为研究不同给药方式下,恩诺沙星在中华草龟体内的药物代谢动力学规律,实验选取20只健康中华草龟,每组10只,随机分为2组,分别为肌注组和胃插管强制口服组,给药量均为10 mg/kg,应用液相色谱—串联质谱法测定数据,Kinetic 4.4软件的非房室模型分析药时数据。胃插管强制口服给药主要药动学参数为C_max7.49 μg/mL、T_max 12 h、T_1/2λz为99.85 h、AUC_0-∞为531.67 μg/(mL·h);肌注给药主要药动学参数为C_max 5.85 μg/mL、T_max 4 h、T_1/2λz为30.42 h、AUC_0-∞为193.6 μg/(mL·h)。2种不同给药条件下,肌注恩诺沙星比胃插管强制口服更快达到最高血药浓度,表明肌注恩诺沙星在中华草龟体内吸收更快;胃插管强制口服给药的C_max和AUC均高于肌注给药,表明胃插管强制口服恩诺沙星在中华草龟体内吸收更完全,分布更广泛,效果更好。恩诺沙星在中华草龟体内消除缓慢,滞留时间(MRT)长,胃插管强制口服给药MRT显著长于肌注给药,其血药浓度保留时间长,效果更持久。研究表明,需较快达到疗效时,建议肌注恩诺沙星;需持续给药且龟体代谢状态正常时,建议胃插管强制口服恩诺沙星;恩诺沙星在胃插管强制口服给药条件下,在中华草龟体内血药浓度高、保留时间长、生物利用度高,更适宜作为中华草龟个体疾病治疗的给药方式。     

Feasibility of using rare earth elements (REEs) to mark and identify escaped farmed Atlantic salmon Salmo salar L.

A series of rare earth elements (REEs) were evaluated as potential markers in scales of Atlantic salmon 1⁺ and 0⁺ smolt by addition of these elements to the feed. Chlorides of 5 REEs were tested, alone or in combination: lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd) and dysprosium (Dy). The labelled‐diets contained 125 or 250 mg REE kg^?1 feed and were administered for up to 10 weeks. Fish scales were collected and analysed by inductively coupled plasma‐mass spectrometry (ICP‐MS) before start, 5 and 10 weeks after labelling started and finally 2 or 4 months after marking. The results demonstrated that the five tested elements were clearly incorporated into the scales of the fish fed the supplemented diets. Uptake and concentrations increased gradually during the feeding period, and no principal differences were found between the two smolt types. Combining 2 REEs in the same feed did not affect the incorporation of either of them. After a ‘dilution’‐period of 2–4 months the concentrations of all markers (except La) in the scales of the treated‐fish were still significantly higher than those of the untreated‐fish. The background concentrations of the tested markers and some other elements were also measured and found to give interesting information.     

Qualitative modifications of humic acid-like and core-humic acid-like during high-rate composting of pig faeces amended with wheat straw

Humic acid-like (HA-like) and core-humic acid-like (core-HA-like) were characterized during the high-rate composting process by CP-MAS ¹³C NMR, pyrolysis-gas chromatography (GC)/mass spectrometry (MS), and elemental analysis. Results obtained indicated that humification proceeded through a relative concentration of aromatic fractions due to the faster degradation of the O-alkyl and alkyl fractions. Core-HA-like, after purification of the parent material, showed a large reduction of the O-alkyl fraction in terms of HA-like. We concluded that HA-like consisted of refractory organic molecules, such as lignin and biopolymers, which formed a stable structure (core-HA-like) coated with degradable material associated with the core by weak physical association, ether or ester bounds.