环境低温对H9N2亚型禽流感病毒感染小鼠致病性的影响

旨在分析环境低温对H9N2亚型禽流感病毒感染小鼠致病性的影响。选用120只6~8周龄,雄性SPF BALB/c小鼠,随机分为4组:无感染常温组(PBS处理,(20±2)℃)、无感染低温组(PBS处理,(10±2)℃)、H9N2病毒感染常温组(H9N2处理,(20±2)℃)、H9N2病毒感染低温组(H9N2处理,(10±2)℃)。在感染后观察感染小鼠发病14 d内的临床症状、存活率,肺脏病变程度、肺部细胞因子含量的动态变化以及肺脏病毒载量。结果表明:1)与H9N2病毒感染常温组小鼠相比,环境低温处理的H9N2病毒感染小鼠临床症状明显加重,其中体重明显降低,但差异不显著(P>0.05),存活率由95%降为67%,肺水肿程度和肺组织病变程度更加严重;2)H9N2病毒感染低温组与H9N2病毒感染常温组相比,肺脏TNF-a和IL-1β含量无显著差异,但均显著高于2个无感染组(P<0.01);3)H9N2病毒感染低温组与H9N2病毒感染常温组相比,肺组织病毒载量显著增加(P<0.01)。综上,环境低温明显增加了H9N2亚型流感病毒对小鼠的致病性,为进一步揭示环境低温与流感病毒感染之间的关系,以及采取有效的预防措施提供数据基础。     

中华鲟小水体养殖试验初报

在7.2m^3水泥池中，采和人工配制饲料对中华鲟（Acipenser sinensis)进行养殖试验，通过4个阶段180d的养殖，鱼尾均增重1817.5g,日均增重分别为8．5，8．9，9．2，12．3，饲料系数为1．96，2．12，2．33，2．21。中华鲟在水水温处于8－29，1℃时均可摄食生长，利用人工配制养殖中华鲟的前景乐观。     

丽江猪屠宰、胴体组成及肉质特性研究

丽江猪是近年在云南丽江新发现的猪种遗传资源,笔者对丽江猪的体型外貌、体尺、胴体组成及肉质性状等种质特性进行测定。结果：丽江猪公、母猪体重分别为（60.71±4.93） kg和（54.11±12.88） kg,体高分别为（65.72±6.11） cm和（64.66±6.10） cm,体长分别为（105.64±9.61） cm和（102.18±10.51） cm,眼肌面积分别为（24.13±6.08） cm2和（23.62±6.16） cm2,瘦肉率分别为（40.09±3.17）%和（41.71±5.05）%,失水率（19.50±5.03）%,粗脂肪（15.71±4.68）%,pH（6.18±0.13）,大理石纹（3.6±0.57）。丽江猪肉质各项指标均符合优质肉范畴,属于肉脂兼用型猪种,且含有丰富的脂肪酸,具有较高食用价值。     

Nanoparticle for siRNA delivery and its pancreatic cancer targeting ability

AIM: To synthesize a safe, efficient and targeted nanoparticulate carrier for siRNA delivery to pancreatic cancer cells. METHODS: Iron oxide nanocrystal with carboxylic acid group-polyethyleneimine (IONP-PEI) was synthesized and investigated as a nonviral carrier of siRNA to the pancreatic cells. The size, surface and charge using zeta potential were characterized. The perfect charge ratio between amino groups of IONP-PEI and phosphate groups of siRNA (N/P) was determined by the transfection efficiency detection, gel retardation assay and MTS assay. An antibody-directed nonviral vector, scFvCD44v6-IONP-PEI nanoparticle attaching to the cancer-associated CD44v6 single-chain variable fragment, was constructed as a cancer-targeting nanocarrier for siRNA delivery. Prussian blue staining and immunofluorescent staining were performed to detect the distribution of scFvCD44v6-IONP-PEI/siRNA complexes in the cells. The transfection efficiency, fluorescence intensity and the expression of KRAS at mRNA and protein levels in the cells transfected by IONP-PEI/siRNA and scFvCD44v6-IONP-PEI/siRNA were detected by flow cytometry, fluorescence microscopy, real-time PCR and Western blotting, respectively. RESULTS：The mass ratio of IONP to PEI was 0.75. The suitable ratio of N/P was 20. The averaged size and surface zeta potential of IONP-PEI/siRNA in deionized water were (51.3±2.2)nm (diameter) and (21.73±8.07)mV,respectively. Red fluorescence was seen in both targeting and nontargeting groups, which clearly revealed the intracellular distribution of siRNA and delivery agents. Transfection efficiencies in targeting and nontargeting groups were (89.75±1.81)% and (59.87±4.52)%, respectively. Down-regulation of the KRAS mRNA in Panc-1 cells transfected with siKRAS by scFvCD44v6-IONP-PEI and IONP-PEI was up to (34.02± 6.15)% and (51.09±6.70)%, respectively. The protein level of KRAS was lower in targeting group than that in nontargeting group. CONCLUSION：scFvCD44v6-IONP-PEI is a safe and efficient nanoparticulate carrier for gene delivery. It is more effective to transfer siRNA into the cells and mediate gene silencing effect in vitro than the nontargeting group.     

Transwell contact co-culture promotes growth and differentiation of single-dissociated iPSCs

AIM：To investigate the promoting role of Transwell contact co-culture system in the growth and differentiation of single-dissociated induced pluripotent stem cells (iPSCs). METHODS：Bovine corneal endothelial cells (CECs) at passage 1~2 (P1~2) were seeded on the underside of Transwell inserts placed into culture plates and were cultured in 37 ℃ and 5% CO2 for 8 h. Accutase digestion and 40 μm filter process disaggregated colony-aggregated iPSCs into single-dissociated iPSCs, and the cells were seeded on the inside of Transwell inserts with CECs in medium of mTeSR1 for 3 d and then in low-glucose DMEM supplemented with 10% FBS for 2 weeks. The characteristics and differentiation markers were evaluated by real-time fluorescence quantitative polymerase chain reaction (qPCR), immunofluorescence staining, live & dead cell staining and alkaline phosphatase (ALP) staining. The group of iPSCs cultured in conventional medium was used as control group 1. The group of single-dissociated iPSCs co-cultured with CECs was set as experimental group, while single-dissociated iPSCs without co-culture were as control group 2. RESULTS：The bovine CECs showed typical hexagonal cobblestone shape. iPSCs showed colony-like growth, while became single-dissociated cells after Transwell contact co-culture with bovine CECs for 3 d. The single-dissociated iPSCs positively expressed the undifferentiated markers, Nanog and Oct4. The mRNA expression levels of Nanog, Oct4 and Sox2 between experimental group and control group 1 were both positive and had no statistical significance difference (P>0.05). The dead cells in experimental group decreased significantly, and there was statistically significant difference compared to control group 2 (P<0.01). After 14 d of induced differentiation co-culture, the single-dissociated iPSCs showed rather uniform polygonal morphology, increased dimension and no obvious colony existence. Negative ALP staining, positive immunofluorescence staining for ZO-1, AQP1 and CD31, and negative for CD34 and CD133 were also observed. The results of qPCR showed that the mRNA expression of Oct4, Nanog and Sox2 significantly decreased, and had statistically significant difference compared with control group 1 (P<0.01). CONCLUSION：When co-cultured with bovine CECs, iPSCs morphologically changed to endothelial-like cells and expressed some markers of CECs. Transwell contact co-culture system not only enhances the growth of single-dissociated iPSCs, but also promotes their differentiation.     

北方蚕区第五批桑品种鉴定（山西点）试验报告

2011—2013年,山西省蚕业科学研究院对北方蚕区3个单位7个桑品种进行了鉴定。田间调查和实验室鉴定结果表明,7个参鉴桑品种生长势强、产叶量高,品质特性良好,在形态特征和生物学特性方面与对照存在不同程度的差异,而昌盛、9330两个品种的表现比较突出。     

稻瘟病菌MoYpt5蛋白互作网络预测

稻瘟病菌(Magnaporthe oryzae)共有11个假定的Rab蛋白家族成员,本文选取了MoYpt51(MGG_06241)和MoYpt52(MGG_01185)进行了生物信息学分析。通过搜索多个大型蛋白互作数据库和文献,共得到数百个与核心蛋白互作的蛋白和互作对。利用信息处理技术和高效制图平台将这些蛋白互作对构建成互作网络,得到若干个具有生物学意义的模块。结果表明,筛选得到的互作蛋白中,有的参与了蛋白降解的泛素途径(MGG_04053等)、囊泡介导的蛋白胞内运输(MGG_01238等),有的在蛋白、染色体的组装和修饰等过程(MGG_03677等)起重要作用。大部分假定互作蛋白定位于细胞质和质膜上,为其与目标蛋白互作提供了空间可能性。     

风沙黄土区排土场不同恢复类型植物群落与土壤种子库特征

风沙黄土区排土场作为一种人造生态系统,自然条件恶劣,土壤贫瘠,植被恢复困难。为了探明有效的人工促进植被恢复措施,采用植被调查与种子库萌发试验相结合的方法,通过研究不同植被类型地上植物群落与土壤种子库特征及二者的关系,探讨了其植被恢复效益及潜力。结果表明:研究区人工植被恢复下地上植物群落中共47种植物,分属16科40属,土壤种子库共14种植物,分属5科13属,其中均以禾本科、菊科、藜科、豆科占主导地位; 灌木植被的地上植被和土壤种子库的物种多样性均表现为最优; 地上植被与土壤种子库密度的变化范围为88.48～495.47株/m²,74.74～1422.91粒/m²,且均在草地类型下最大。土壤种子库和地上植被的相似性普遍较低,相似性系数仅为0.16～0.23。因此,风沙黄土区排土场的植被恢复与重建需要加强保护与管理,可以考虑构建以草灌配置为主的人工植被恢复模式,保障群落的恢复潜力,并提高群落多样性与稳定性,亦可考虑引入外源种子库提升群落恢复的潜力。     

3种固沙材料与风沙土复配后土壤改良效应及其质量评价

揭示风沙土复配其他固沙材料后的土壤质量变化及改良效应对研发防沙固沙新材料具有重要的指导意义。以乌兰布和沙漠风沙土为对照(CK),采用粉煤灰、脱硫石膏、牛粪3种固沙材料按干质量比15%,25%与风沙土进行复配,研究了不同材料与配比复配土壤对机械组成、养分含量、金属含量的改良效应,运用最小数据集构建、隶属度计算与土壤质量指数计算等方法评价了复配土壤质量。结果表明:(1)不同复配土壤的机械组成均发生了改变,粉粒、粗砂等难蚀、较难蚀颗粒体积分数增加,易蚀的细砂体积分数降低。(2)除石膏复配土壤各养分指标与CK相近外,其余复配土壤有机碳含量较对照提高1.2～10.4倍,牛粪复配土壤在提高养分含量方面显著高于其他材料; 以牛粪和石膏为材料的复配土则分别增加或降低了沙土中的金属含量,而粉煤灰复配土的Cr,As较CK下降。(3)不同复配土对土壤理化性质作用影响各异,粉煤灰在粒径改良方面较优,牛粪在养分提升方面效果显著,石膏在重金属含量降低方面作用明显,粉煤灰石膏混合复配则综合了二者的优势。(4)3种固沙材料均在不同程度上提高了复配土壤的质量,但复配土壤质量仍处于较低水平。综上,不同复配比例组合中,15%粉煤灰复配土壤质量得分指数最高为0.545,对风沙土的改良效果最好,其次为15%脱硫石膏(0.537),15%牛粪(0.506)和25%粉煤灰与脱硫石膏(0.484)复配组合。     

In vitro Selection of DNAAptamers and Fluorescence-Based Recognition for Rapid Detection Listeria monocytogenes

Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides. Systematic evolution of ligands by exponential enrichment （SELEX） was used to select and PCR amplify DNA sequences （aptamers） capable of binding to and detecting Listeria monocytogenes, one of the major food-borne pathogens. A simplified affinity separation approach was employed, in which L. monocytogenes in exponential （log） phase of growth was used as the separation target. A fluorescently-labeled aptamer assay scheme was devised for detecting L. monoeytogenes. This report described a novel approach to the detection of L. monocytogenes using DNA aptamers. Aptamers were developed by nine rounds of SELEX. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. One of aptamers named e01 with the highest affinity was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. This study has proved the principle that the whole-cell SELEX could be a promising technique to design aptamer-based molecular probes for dectection of pathogenic microorganisms without tedious isolation and purification of complex markers or targets.