小麦苗枯病菌的ITS分析及PCR检测

 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。     

细胞因子对小尾寒羊胎盘成熟的影响

为了研究细胞因子对小尾寒羊胎盘成熟的影响 ,本实验采用液相竞争法和平衡法 ,对小尾寒羊空怀期 (n=5 )和妊娠期 (n=13)第 85天、10 5天、12 5天、14 0天和 15 0天 (足月 )时的血清白细胞介素 - 1β(IL - 1β)、白细胞介素 - 6 (IL - 6 )和肿瘤坏死因子(TNF)的含量分别进行检测。结果表明 ,IL - 1β和 IL - 6含量在 12 5天时与对照组间差异极显著 (P<0 .0 1) ,TNF含量在各时期与对照组差异均显著 (P<0 .0 1或 P<0 .0 5 )。随着妊娠过程的进展 ,三种细胞因子含量逐渐增加 ,12 5天时达到最高值 ,分别为 0 .390± 0 .196 ng/ ml,5 79.8± 15 2 .8pg/ ml和 2 .348± 0 .396 ng/ ml。而后逐渐下降 ,到足月前略有回升 ,但仍低于 12 5天时的最高值。由此看出小尾寒羊在妊娠期间 ,细胞因子与妊娠之间有着密切的关系 ,前者对于维持妊娠起到重要作用。由于 14 0天至足月3种细胞因子基本稳定 ,这表明胎盘于 14 0天左右达到成熟 ,IL- 1β和 IL- 6与启动分娩有关 ,而 TNF可能参与小尾寒羊分娩的启动。     

牛胎儿成纤维细胞分离与体外培养

本研究以 4 5d牛胎儿为材料 ,使用 0 .2 5 %胰酶 + 0 .0 4 %EDTA消化液 ,分离得到牛胎儿成纤维细胞 ,体外培养传代至第 2 8代。本实验描绘出牛胎儿成纤维细胞生长曲线 ,并发现α MEM是一种适合牛胎儿成纤维细胞生长的培养基。分离得到的牛胎儿成纤维细胞传至第 1 8代时核型仍然正常 ,冷冻、解冻后细胞可继续传代     

H5N1亚型禽流感病毒核蛋白基因的克隆及分子进化分析

用RT-PCR法,以1株h5N1亚型禽流感病毒RNA为模板,扩增了NP全基因cDNA.将cDNA克隆于pMD18-T载体,并对其进行测序.测序结果表明所克隆的1542个核苷酸的片段包含了核蛋白完整阅读框架,其中编码区为1497个核苷酸,编码498个氨基酸.将其序列与6株A型流感病毒NP基因序列进行比较,各毒株之间NP基因核苷酸序列同源性为92.5%～95.9%,编码的氨基酸同源性为97.0%～98.4%.本研究通过分子进化分析揭示了各毒株间的亲源关系.     

VEGF在绵羊生殖管道中的表达规律

以不同发情周期雌性绵羊子宫、输卵管为研究对象,采用免疫组织化学技术,针对血管内皮生长因子（VEGF）在绵羊子宫、输卵管的表达、定位和变化规律进行了检测,同时应用相关图像分析软件对抗原染色强度进行了定量分析。结果表明：输卵管在发情0～15d,VEGF表达量在第9天达到峰值后经历波动逐渐下降过程,输卵管内膜上皮细胞是VEGF抗原的主要靶细胞;而子宫角在发情0～15d,VEGF表达量在第5天达到峰值后经历波动逐渐下降过程,子宫内膜固有层及腺体周围细胞为VEGF抗原的主要靶细胞。该研究结果为绵羊生产中进一步提高受胎率和妊娠率及频密产羔等技术的应用提供了科学依据。     

双孢菇菌渣基质的制备及其栽培黄瓜的效果比较

以双孢菇菌渣为研究对象,通过设置不同发酵菌、蔗糖和尿素的添加量,测定分析不同处理菌渣发酵后的容重、总孔隙度、通气孔隙度、持水孔隙度、电导率(EC)值及pH;不同处理发酵菌渣按照2∶1的体积比与蛭石混配后栽培黄瓜,以草炭和蛭石2∶1的体积比为对照,测量分析黄瓜的株高、糖含量和产量,以期探索菌渣发酵添加物的合理配比.结果 表明:1000 kg菌渣加入8 kg麦麸发酵菌、6 kg蔗糖、6 kg尿素,经过20～25 d后能够充分发酵,发酵后的菌渣基质理化性质符合作物栽培的要求,用于黄瓜栽培,其生长速度、糖度和产量最高,且显著超过了对照.     

不同粗纤维水平饲粮对烟台黑猪肥育性能及胴体肉品质的影响

选择2014年春季出生的生长发育良好、胎次相近、体重35 kg左右的烟台黑猪仔猪95头为研究对象,随机分为3组,低纤维水平组31头饲喂低纤维水平饲粮,中纤维水平组34头饲喂中等纤维水平饲粮,高纤维水平组30头饲喂高纤维水平饲粮。每组3个重复,每个重复10~12头。结果表明:不同粗纤维水平饲粮对烟台黑猪35~75 kg生长期和75~95 kg肥育期的平均日增重影响显著。其中,生长期平均日增重以低纤维水平组最高,分别比中纤维水平组和高纤维水平组提高14.91%(P0.05)、23.05%(P0.01);肥育期平均日增重以中纤维水平组最高,分别比低纤维水平组、高纤维水平组提高20.96%(P0.05)、5.62%(P0.05)。生长期和全期料重比以低纤维水平组最低即饲料报酬最高,分别比中纤维水平组、高纤维水平组降低6.59%、15.51%、2.85%、6.14%。饲粮粗纤维水平对烟台黑猪胴体性能影响不显著,对肉质性状中的a值及干物质含量影响显著。a值、干物质和肌内脂肪含量都以低纤维水平组最高,分别比中纤维水平组和高纤维水平组提高9.75%(P0.05)、13.72%(P0.05)、6.09%(P0.05)、8.31%(P0.05)和110%(P0.05)、122.35%(P0.05)。不同粗纤维水平饲粮对烟台黑猪背最长肌的酪氨酸和苯丙氨酸影响显著,酪氨酸和苯丙氨酸含量都以高纤维水平组最高,分别比低纤维水平组和中纤维水平组提高10.00%(P0.05)、4.05%(P0.05)、11.90%(P0.05)、5.62%(P0.05)。不同粗纤维水平饲粮对烟台黑猪背最长肌亚油酸含量影响显著,对其他3种脂肪酸影响不显著。亚油酸含量以高纤维水平组最高,比低纤维水平组、中纤维水平组分别提高35.14%(P0.05)、4.74%(P0.05),其次是中纤维水平组较高,比低纤维水平组提高29.02%(P0.05)。综合分析,烟台黑猪适宜的饲粮粗纤维水平生长期为10%,肥育期为12%。     

2013 年国家鉴定的辣椒品种（一）

师研1 号、苏椒18 号、中椒0808 号、冀研16 号、京甜1 号、沈研15 号、湘研808、金田8 号、东方168、粤研1 号     

Preliminary study of patients with chronic mountain sickness by GC-TOF-MS based serum metabolomics analysis

AIM: To evaluate specific metabolomics profiles in the serum of patients with chronic mountain sickness (CMS) and to explore the potential metabolic biomarkers in the native Tibetans living on the Qinghai-Tibet Pla-teau.METHODS: A gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) approach as a metabolomics technique was used to evaluate metabolic differences. The native Tibetan CMS patients (n=10) and healthy Tibetan controls (n=10) were enrolled from YuShu in Qinghai province in this study. The serum samples were collected and analyzed by GC-TOF-MS coupled with a series of multivariate statistical analyses such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA).RESULTS: The intergroup differences between CMS patients and control subjects have been observed. A list of differential metabolites and several top altered metabolic pathways have been identified. The levels of fumaric acid, an intermediate in the tricarboxylic acid (TCA) cycle, and inosine were highly upregulated in the CMS patients, suggesting a greater effort to hypoxic adaptation in high elevation area. Other differential metabolites, such as methyl phosphate, 2-ketoadipate, lyxose and phytanic acid were also identified. Importantly, the differential metabolites possessed higher area under the ROC curve (AUC) values, indicating an excellent clinical ability for the prediction of CMS. Increased levels of amino acids (isoleucine, glycine, serine, L-cysteine, citrulline and trimethyllysine) were detected in CMS group, yet significantly decreased levels of sulfuric acid, oxamic acid, lyxose and glutamine were also detected in CMS group than those in control group. At the same time, the levels of ribose and glucose-1-phosphate were markedly elevated in CMS group (P<0.05).CONCLUSION: The metabolic activities are significantly altered in the serum of CMS patients. High altitude hypoxia may act on the disturbed glucose metabolism and amino acid metabolism in part of the Tibetan triggered by CMS.     

F-box domain is involved in regulating the proliferation of MCF-7 cells by FBXO39 protein

AIM: To investigate the effect of F-box domain on the regulation of MCF-7 cell proliferation by FBXO39 protein. METHODS: The effect of F-box domain on the localization of FBXO39 protein in the MCF-7 cells was investigated. MCF-7 cell cDNA library was used as the template resource. The full-length cDNA sequence of FBXO39 was amplified by PCR method and subcloned into eukaryotic expression vector pEGFP-C2. The pEGFP-FBXO39ΔF (F-box domain deletion mutation) plasmid was successfully constructed with the template resource of pEGFP-FBXO39 plasmid. The recombinant plasmids were transfected into the MCF-7 cells, and then the expression of FBXO39 and FBXO39ΔF were determined by Western blot. The cellular localization of FBXO39 and FBXO39ΔF were observed by confocal microscopy. The localization of endogenous FBXO39 in the MCF-7 cells was detected by immunofluorescence staining. In addition, MTT and EdU assays were used to measure the cell proliferation, flow cytometry was used to measure the cell cycle distribution, and immunohistochemical staining was used to observe the expression of FBXO39 in the breast cancer and para-carcinoma tissues. RESULTS: The eukaryotic expression vector pEGFP-FBXO39 and pEGFP-FBXO39ΔF were constructed successfully. F-box domain had no effect on the cell localization of FBXO39. FBXO39 promoted MCF-7 cell proliferation but FBXO39ΔF did not. FBXO39 was highly expressed in the breast cancer tissues. CONCLUSION: F-box domain had no effect on the cellular localization of FBXO39 protein. However, it plays an important role in the biological function of FBXO39. FBXO39 may be related to breast cancer tumorigenesis.