SNAT2介导蛋氨酸对牛乳腺上皮细胞增殖与自噬的调节作用

钠离子依赖性中性氨基酸转运体2（SNAT2）是一种氨基酸转运蛋白,可转运中性氨基酸,广泛分布于多种细胞中。氨基酸既可作为蛋白质合成的底物,也是调节细胞新陈代谢的关键信号分子,但SNAT2是否介导氨基酸调节BMECs增殖和自噬尚未见报道。本研究利用CASY细胞计数和Western blotting技术检测SNAT2过表达和siRNA干扰后牛乳腺上皮细胞（BMECs）增殖情况以及SNAT2对自噬标志蛋白LC3-Ⅰ/Ⅱ表达量的影响,并利用免疫荧光检测细胞自噬斑点（LC3-Ⅱ）变化。结果显示,SNAT2过表达时,p-PI3K、p-mTOR和Cyclin D1表达量增加,反之,p-PI3K、p-mTOR和Cyclin D1表达量下降。SNAT2抑制时,LC3-Ⅱ表达量增加,免疫荧光检测自噬斑点增多。添加自噬增强剂海藻糖（trehalose,Tre）和蛋氨酸（methionine,Met）后,与单一添加Tre组相比,Met+Tre组p-mTOR表达量增加,LC3-Ⅱ表达量降低,胞浆内绿色自噬斑点减少;添加Tre和Met并抑制SNAT2时,p-mTOR表达量下降,LC3-Ⅱ表达量增多,胞浆内绿色自噬斑点增加。以上结果表明,SNAT2可介导Met通过调控PI3K-mTOR/Cyclin D1信号通路调节BMECs的增殖与自噬。     

Effect of the β‐carotene oxygenase 2 genotype on the content of carotenoids,retinol and α‐tocopherol in the liver,fat and milk of rabbit does,reproduction parameters and kitten growth

Mutations in the β‐carotene oxygenase 2 (BCO2) gene can impair the function of the enzyme that breaks down carotenoids. As a result, gradual accumulation of unoxidized carotenoids in animal tissues gives them a yellow colour. The aim of the study was to determine the content of carotenoids, retinol and α‐tocopherol in the liver, fat and milk of rabbit does with three different genotypes determined by AAT‐deletion mutation at codon 248 of the BCO2 gene and to find out whether differences in the concentrations of the above compounds in the tissues and milk of the does affect reproduction parameters and the rearing rate of kittens. The experimental materials comprised 36 does, 12 of each genotype of the BCO2 gene, with their litters. Females with their litters were placed in individual cages, on deep litter. Between days 7 and 13 of lactation, samples of milk were collected from the does. The kittens stayed with their mothers until 35 days of age. After weaning, the does were sacrificed. Tissue samples of liver and perirenal fat were collected for chemical analyses. Additionally, based on samples taken from one female, RNA expression levels were determined from the mammary gland and liver, adipose tissue and skin. It was found that homozygous does with deletion at codon 248 of the BCO2 gene were characterized by considerably higher concentrations of xanthophylls and beta‐carotene in the liver, adipose tissue and milk than does with the remaining genotypes. However, the differences in the content of the above compounds in milk had no influence on litter weight or the number and rearing rate of kittens. Additionally, RNA expression of the BCO2 gene was found in the mammary tissue of lactating doe and its level was similar to those noted in the liver and adipose tissue.     

Ascorbic acid ameliorates dysregulated folliculogenesis induced by mono-(2-ethylhexyl)phthalate in neonatal mouse ovaries via reducing ovarian oxidative stress

Phthalates, including di-(2-ethylhexyl)phthalate (DEHP), are common industrial chemicals in the environment. Recent evidence indicates that DEHP and its active metabolite mono-(2-ethylhexyl)phthalate (MEHP) negatively modulate reproductive functions and induce reactive oxygen species. Ascorbic acid (AA) is a dietary requirement for primates, and it acts as a potent free radical scavenger to protect tissues against oxidative stress. In this study, to investigate the toxic effects of MEHP on the follicle development and the beneficial role of AA, neonatal mouse ovaries were treated with different concentrations of MEHP with or without AA for 6 days. Then, the follicle constitution and oxidative status were compared in different groups. Results showed MEHP accelerated primordial follicle recruitment by increasing the percentage of primary and secondary follicles and decreasing the percentage of primordial follicles in the ovaries. Moreover, MEHP-induced ovarian oxidative stress by significantly increasing malondialdehyde (MDA) concentration and the expression of GSS and SOD1. When ovaries were co-administrated with MEHP and AA, follicle constitution was normalized, and the oxidative status was significantly decreased. These results suggested that AA ameliorated MEHP-induced ovarian oxidative stress and follicular dysregulation, which attested the clinical significance of AA for ovary protection in the case of MEHP exposure.     

氨基色谱柱测定蜂蜜中4种糖方法研究

《中国药典》(2015年)蜂蜜中规定:用Prevail Carbohyrate ES色谱柱,以乙腈-水(75:25)为流动相,示差折光检测器测定蜂蜜中的果糖、葡萄糖、蔗糖及麦芽糖。此次试验证明用普通氨基色谱柱替代Prevail Carbohyrate ES色谱柱,待测组分与其他组分分离良好,精密度试验满足要求,回收率试验结果大于93%,结果准确、可靠,普通氨基柱可替代Prevail Carbohyrate ES色谱柱,用于蜂蜜中糖类物质的测定。     

REGIONAL BINDING INDEX OF THE RADIOLABELED SELECTIVE 5-HT2A ANTAGONIST 123I-5-I-R91150 IN THE NORMAL CANINE BRAIN IMAGED WITH SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY

The pattern of the specific 5-HT2A (5-hydroxytryptamine 2A receptor) antagonist 123I-5-I-R91150 was measured in 10 healthy dogs without neurologic and behavior abnormalities. Eight cortical regions (left and right fronto-, temporo-, parieto-, and occipitocortical area), one global subcortical region (including the thalamic system) were compared with a reference region lacking receptors; that is, the cerebellum. The 123I labeled radioligand was injected intravenously 100-200 minutes before acquisition. Both transmission and emission data were obtained with a triple head gamma camera equipped with high-resolution fanbeam collimators. The emission data were corrected for scatter and attenuation. To delineate different cerebral regions more accurately, the regions of interest (ROI) defined in a former study on brain perfusion measured with 99mTc-ethyl cysteinate dimer (ECD) in the same dogs were used. The co-registration of the 99mTc-ECD and the 123I-5-I-R91150, obtained from each dog, was realized with the help of corresponding transmission maps. By normalizing each regional cerebral activity to the activity observed in the cerebellum, the regional radioactivity (binding index) could be relatively quantified. Highest brain uptake was noted in the frontocortical brain areas (right: 1.85, left: 1.89), followed by the temporocortical region (right: 1.58, left: 1.56). Least uptake was noted in the more caudal and middle brain regions [occipito- (right: 1.46, left: 1.41), parietocortical (right: 1.30, left: 1.26), and striatal region (1.19)]. No gender nor age influence was noted in this series. The 123I labeled serotonin-2A receptor ligand seems to have similar cortical binding in the normal canine brain, as shown in humans and other animal species. A frontocortical to occipitocortical (rostrocaudal) binding index gradient was identified within the dog, which has not been seen in imaging studies from humans and other animal species. The significance of these results will need further investigation. This normative data can be used to compare regional brain uptake of the 123I-radioligand to dogs with behavioral disorders related to the serotonergic system, in future studies.     

早期妊娠猕猴孕激素水平的检测

利用全自动化学发光免疫分析仪检测正常未妊娠的与不同孕期猕猴血清中雌二醇及孕酮的分泌水平。结果表明孕5d、10d、25d、30d血清孕酮比正常猕猴的分泌水平有非常明显的上升（P〈0.01）,孕15d也有显著的上升（P〈0.05）;孕15d、20d的雌二醇分泌水平比正常组有非常显著升高（P〈0.01）,而孕5d、10d、2...     

通用型流感疫苗研究进展

疫苗免疫是目前防控流感病毒感染最有效的策略。现有流感疫苗在不同亚型间的交叉保护力较差,并且由于HA和NA蛋白存在抗原漂移和转换现象,极大地影响了现有疫苗的免疫保护效果,甚至导致疫苗免疫失败。而动物模型研究结果显示,基于流感病毒基因组保守区域的疫苗能够对不同型或亚型的流感病毒产生广谱的交叉保护,本文主要介绍基于M2e、HA和NP的通用型流感型疫苗研究进展。     

猪传染性胸膜肺炎放线杆菌和猪圆环病毒2型二重实时荧光定量PCR检测方法的建立

以胸膜肺炎放线杆菌(App)apx ⅣA基因和猪圆环病毒2型(PCV-2)ORF2基因为靶基因,分别设计特异性引物和探针,建立App和PCV-2的二重荧光PCR方法。App引物扩增的目的片段大小为132bp,PCV-2为169bp。建立的方法可在同一反应体系中同时对App和PCV-2进行定量检测鉴别,其敏感性分别为1×101拷贝和1×100拷贝,对其他猪病病原核酸的检测为阴性。利用该方法对10份人工感染临床样品进行检测试验,其结果符合率达100%,说明该方法可用于对临床样品中App和PCV-2的检测。     

Closed system anaesthesia in dogs using liquid sevoflurane injection; evaluation of the square-root-of-time model and the influence of CO2 absorbent

Objective To determine whether predictable alveolar concentrations of sevoflurane are reliably produced in dogs when liquid sevoflurane is injected into closed circuit breathing systems, as calculated by Lowe's square‐root‐of‐time anaesthetic uptake model, and to confirm the validity of the model using soda lime and calcium hydroxide lime. Study design Prospective clinical study. Animals Eleven healthy dogs with a mean body mass of 34 ± 9 kg scheduled for pelvic limb orthopaedic surgery. Materials and methods Following pre‐anaesthetic medication, anaesthesia was induced with propofol and maintained with sevoflurane in a closed circle system. Epidural anaesthesia was performed with morphine and bupivacaine. Liquid sevoflurane was injected into the circuit by syringe, using dosages and time intervals derived from Lowe's square‐root‐of‐time anaesthetic uptake model. The target alveolar concentration chosen was 1.1 × MAC (2.6% end‐tidal sevoflurane). Either soda lime (group S; n = 6) or calcium hydroxide lime (Amsorb; group A; n = 5) were used for CO₂ absorption. Sevoflurane concentration and the respiratory gas composition were measured with an infrared gas analyser. Results End‐tidal sevoflurane concentrations were close to the predicted value of 2.6% at 9 minutes (2.53 ± 0.1% group S; 2.60 ± 0.26% group A) and 16 minutes (2.55 ± 0.30 group S; 2.52 ± 0.28% group A) but declined thereafter to reach 50% (group S) and 64% (group A) of the predicted value at 121 minutes. There was a constant trend towards higher end‐tidal sevoflurane concentrations in group A but the difference was not statistically significant. Conclusions The square‐root‐of‐time model leads to significantly lower alveolar concentrations than expected, suggesting that the rate of sevoflurane uptake in dogs declines less rapidly than predicted. The use of Amsorb tends to reduce the deviation from predicted concentrations. Clinical relevance The model used in this study provided only an approximate guide to the volume of liquid sevoflurane required. Consequently, the definitive dose schedule must be based on measured anaesthetic concentrations and clinical monitoring.