To characterize the composition of follicular fluids (FF) collected from large and medium sized follicles of domestic pigs,gas chromatography coupled to time-of-flight mass spectrometry (GC/TOF-MS) was used to detect endogenous metabolites in the three-breed crossbred porcine follicular fluids collected from follicles of large (5-8 mm) and medium (3-5 mm) diameters.Finally,metabolite profiles of the follicular fluids were determined by multivariate statistical analysis,such as principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA).The result showed that the metabolic profiles of the large FF and the medium FF were evidently different.The concentrations of 33 metabolites differed from each other due to different follicle sizes (P＜0.05).The concentrations of all these metabolites,except glucose,α-ketoglutaric acid,proline,and glucose l-phosphate,were lower in the large FF,compared with the medium FF;which was mainly associated with the TCA cycle,glycometabolism,amino acid and nucleoside metabolism,based on the biochemical metabolic pathway analysis.There was a distinct shift in follicular metabolic pathway as follicles increased in diameter.Our findings might provide a new clue for study on developmental potential of the oocytes,early embryo development,and regulation of follicular development by means of artificial intervention.

Neuromedin B (NMB), a mammalian bombesin-related peptide, has numerous physiological functions, including regulating hormone secretions, cell growth, and reproduction, by binding to its receptor (NMBR). In this study, we investigated the effects of NMB on testosterone secretion, steroidogenesis, cell proliferation, and apoptosis in cultured primary porcine Leydig cells. NMBR was mainly expressed in the Leydig cells of porcine testes, and a specific dose of NMB significantly promoted the secretion of testosterone in the primary Leydig cells; moreover, NMB increased the expression of mRNA and/or proteins of NMBR and steroidogenic mediators (steroidogenic acute regulatory (STAR), CYP11A1, and HSD3B1) in the Leydig cells. In addition, specific doses of NMB promoted the proliferation of Leydig cells and increased the expression of proliferating cell nuclear antigen and Cyclin B1 proteins, while suppressing Leydig cell apoptosis and decreasing BAX and Caspase-3 protein expression. These results suggest that the NMB/NMBR system might play an important role in regulating boar reproductive function by modulating steroidogenesis and/or cell growth in porcine Leydig cells.

The aim of our study was to investigate whether miR-15a has effect on the development of porcine follicular by targeting on BD-NF or not.We extracted 3-5 mm healthy,3-5 mm atretic,more than 5 mm healthy follicular and found that the level of miR-15a in the diameter of 3-5 mm of atretic follicular is more than the healthy follicular by using of qPCR.BDNF and its high-affinity receptor TrkB were stronger expressed than those apoptotic cell which approaching follicular antrum by using immunohistochemistry.After isolating and cultivating porcine follicular cell,overexpression of miR-15a could significantly upregulate BDNF's mRNA and downregulate bcl-2's mRNA without significant difference.Inhibition of miR-15a could significantly downregulate the expression of BDNF and bcl-2's mRNA.Western-Blot results showed that overexpression of miR-15a could obviously downregulate BDNF protein level,while inhibition of miR-15a can significantly upregulate BDNF level.The results suggest that miR-15a may be involved in the development and atresia of porcine follicles via targeting BDNF.

旨在建立猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)抗体胶体金免疫层析检测方法,从而快速地对Mhp的感染和免疫情况进行监测,预防和控制猪支原体肺炎传播.通过柠檬酸三钠还原法制备胶体金颗粒,确定Mhp特异性抗原P46蛋白的最适标记pH和标记浓度并进行标记,作为金标液.将P46蛋白包被检测线T线,抗P46蛋白单抗包被质控线C线,组装检测试纸条.经优化,建立可同时用于Mhp血清抗体和黏膜抗体检测的胶体金免疫层析检测方法.利用该检测方法对76份临床血清样品和40份临床鼻拭子样品进行检测,并与ELISA检测结果进行比较.用该试纸条检测血清样品时,灵敏性、特异性、重复性较好,与商品化Mhp ELISA抗体检测试剂盒符合率为96.1％;该方法检测呼吸道鼻拭子样品时,存在微弱的非特异性反应,检测结果与本实验室建立的sIgA-ELISA检测方法符合率为87.5％.本研究成功建立了Mhp抗体胶体金免疫层析检测方法,整个过程只需10 min.检测血清样品具有较好的特异性和灵敏性,检测鼻拭子样品时特异性还有待提高.

The present study was conducted to examine the short-term calorie restriction effects (CR) on reproduciton in male rats.Twenty adult male Wistar rats were separated to two groups randomly,including calorie restriction group and control group.Controls were raised ad libitum and treatments were raised with 60％ of the food expended by control rats.After seven weeks,the rats were weighed and observed the mental state.Then rats were executed and the testicular tissues were embedded into paraffin stained with hematoxylin and eosin (HE).Immunostaining was carried out with mice polyclonal antibodies to ZO-1.Then,oxidative stress parameters,including MDA,T-SOD and GSH-Px,were measured.Total RNA was extracted from testicular tissues and the expression of mRNA of ZO-1,SOD,CAT,GST and Nrf2 were measured by real-time quantitative PCR.The results showed that the body weight of short-term restricion group was lighter than control group.However,rats had better mental state in short-term calorie restricion group.No significant differences were observed between the short-term calorie restricion group and the control group about the diameter of seminiferous tubule.Rats had good seminiferous tubule and intact basilemma in short-term calorie restricion group.Various kinds of the germ cells were neatly arranged in the seminiferous tubules.There were six to eight layer cells in seminiferous epithelium.The expression of ZO-1 mRNAs was significantly increased in the short-term calorie restriction group compared with the control group,and the positive reaction of ZO-1 protein was observed.MDA was reduced in calorie restricion groups (P＜0.05).However,T-SOD and GSH-Px were increased because of calorie restricion (P＜0.05).The expression of mRNAs of SOD,CAT,GST,Nrf2 were increased by calorie restricion treatment (P＜0.05).The results suggest that short-term calorie restriction can promote tight junctions between testicular cells and improve reproduction through enhancing the ability of antioxidation and decreasing the poisonousness of free radical to cells.

我和在座的班主任一样，每天都在忙碌着、思索着、改进着、提高着。十几年的班主任经历，使我感受概括为“用情育、用心管、用智引”。

水库水属于低温低浊低活性水,采用常规工艺难以净化处理.近年来由于环境变化使水库水的色度、耗氧量提高,进一步增加了水质净化的难度.雨山水厂在原来传统水处理工艺的基础上,针对水库水质的特点,对取水、混凝、沉淀和过滤等环节进行了技术改进,取得了良好的效果,在水库水源水处理技术改进方面做了有益的探索.

<正>2004年,国家出台的农机购置补贴政策,对滑县农业的发展起到了决定性的转折作用。在农机购置补贴政策的带动下,滑县农机化很快进入了一个全新的发展时期。滑县,位于河南东北边陲,这里土地广袤、富饶,有"豫北粮仓"之称,以小麦和玉米为主的农作物,现已喜获"十一连增"。10月下旬,滑县已经结束了玉米的收获和小麦的播种工作。近年来,随着粮食产量的逐年攀升,滑县"三夏""三秋"的时间却越来越短。这对于身兼"农""工"身份的现代农

介绍了湖北省通山县第一次水利普查坚持不懈,高质量推进清查登记、台账建设、普查数据采集、数据处理与空间数据标绘等工作的做法和经验.