2型糖尿病大鼠主动脉硬化模型的构建

目的:建立自发2型糖尿病大鼠主动脉硬化动物模型.方法:用2型糖尿病大鼠主动脉硬化动物模型,44只GK大鼠随机分为正常GK对照组、建模组,每组22只,正常Wistar大鼠组22只.期间观察大鼠血糖、尿糖及一般情况变化,8周后,测定各组动物空腹血糖、总胆固醇(total cholesterol,TC)、甘油三酯(triglycerides,TG)、低密度脂蛋白胆固醇(low density lipoprotein choleste rol,LDL-C),同时光学显微镜观察主动脉结构.结果:模型组大鼠的总胆固醇、甘油三酯、低密度脂蛋白胆固醇明显升高,出现主动脉肥大,病理显示明显的主动脉及动脉内膜病变.结论:通过高脂、一氧化氮合成酶抑制剂,可成功构建自发2型糖尿痛大鼠主动脉病变,用作2型糖尿病大鼠大血管病变研究的动物模型.     

重组腺相关病毒介导的大鼠脂联素载体的构建与鉴定

目的:克隆大鼠脂联素基因(Acrp30),并对载有脂联素的重组腺相关病毒(AAV)载体进行构建及鉴定.方法:根据Acrp30的基因序列设计引物,上游引入EcoR Ⅰ位点,下游引入Sal Ⅰ位点,以EX-A0242-M01-Acrp30为模板扩增目的基因.将Acrp30基因克隆入pUC19载体中,获得重组质粒pUC19-Acrp30.然后用EcoR Ⅰ与sal Ⅰ酶切pUC19-Acrp30及质粒pSNAV,回收酶切片段,T4DNA连接酶16℃过夜,转化大肠杆菌DH5a感受肽细胞,筛选阳性克隆获得pSNAV-Acrp30,EcoR Ⅰ/Sal双酶切鉴定,并行全基因测序.结果:PCR电泳及酶切鉴定表明,pSNAV-Acrp30重组成功,基因测序显示装入pSNAV质粒中的Acrp30基因正确.结论:脂联素病毒栽体成功构建,可以满足脂联素基因研究和治疗的需要.     

Eotaxin-3基因多态性与类风湿性关节炎相关性研究

目的:研究湖北地区汉族成人eotaxin-3 77C/T和 2497T/G单核苷酸多态性与类风湿性关节炎易感性之间的关系。方法:用聚合酶链反应-单链构象多态性-四引物聚合酶链反应-限制性酶切的方法进行分析。结果:类风湿关节炎组与对照组eotax- in-3 2497位基因型频率及等位基因的频率差异有统计学意义(P<0.05); 77位基因型频率及等位基因的频率差异无统计学意义(P>0.05)。结论:eotaxin-3 2497T/G多态性跟类风湿关节炎易感性相关。     

Immunization with chlamydial plasmid protein pORF5 DNA vaccine induces protective immunity against genital chlamydial infection in mice

To validate the immune protective efficacy of pORF5 DNA vaccine and to analyze potential mechanisms related to this protection. In this study, pORF5 DNA vaccine was constructed and evaluated for its protective immunity in a mouse model of genital chlamydial infection. Groups of BALB/c mice were immunized intranasally with pORF5 DNA vaccine. Humoral and cell mediated immune responses were evaluated. The clearance ability of chlamydial challenge from the genital tract and the chlamy- dia-induced upper genital tract gross pathology and histopathological characterization were also de- tected. The results showed that the total and the IgG2a anti-pORF5 antibody levels in serum were sig- nificantly elevated after pcDNA3.1-pORF5 vaccination, as were the total antibody and IgA levels in vaginal fluids. pcDNA3.1-pORF5 induced a significantly high level of Th1 response as measured by robust gamma interferon (IFN-γ). Minimal IL-4 was produced by immune T cells in response to the re-stimulation with pORF5 protein or the inactive elementary body in vitro. pcDNA3.1-pORF5-vacci- nated mice displayed significantly reduced bacterial shedding upon a chlamydial challenge and an accelerated resolution of infection. 100% of pcDNA3.1-pORF5 vaccinated mice successfully resolved the infection by day 24. pcDNA3.1-pORF5-immunized mice also exhibited protection against patho- logical consequences of chlamydial infection. The stimulated index was significantly higher than that of mice immunized with pcDNA3.1 and PBS (P<0.05). Together, these results demonstrated that immu- nization with pORF5 DNA vaccine is a promising approach for eliciting a protective immunity against a genital chlamydial challenge.     

Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio)

The aim of the present study was to purify the common native carp growth hormone (ncGH), produce monoclonal antibodies (mAbs) to common native carp growth hormone (ncGH), and further enhance the sensitivity of enzyme-linked immunosorbent assays (ELISA) for ncGH. Additionally, we investigated changes in serum ncGH levels in carps raised in different environmental conditions. The recombinant grass carp (Ctenopharyngodon idella) growth hormone was purified and used as antigen to immunize the rabbit. The natural ncGH was isolated from the pituitaries of common carp. SDS-PAGE and Western blot utilizing the polyclonal anti-rgcGH antibody confirmed the purification of ncGH from pituitaries. Purified ncGH was then used as an immunogen in the B lymphocyte hybridoma technique. A total of 14 hybridoma cell lines (FMU-cGH 1-14) were established that were able to stably secrete mAbs against ncGH. Among them, eight clones (FMU-cGH1-6, 12 and 13) were successfully used for Western blot while nine clones (FMU-cGH 1-7, 9 and 10) were used in fluorescent staining and immunohistochemistry. Epitope mapping by competitive ELISA demonstrated that these mAbs recognized five different epitopes. A sensitive sandwich ELISA for detection of ncGH was developed using FMU-cGH12 as the coating mAb and FMU-cGH6 as the enzyme labeled mAb. This detection system was found to be highly stable and sensitive, with detection levels of 70 pg/mL. Additionally, we found that serum ncGH levels in restricted food group and in the net cage group increased 6.9-and 5.8-fold, respectively, when compared to controls, demonstrating differences in the GH stress response in common carp under different living conditions.     

西方蜜蜂王浆产量与品质性状的配合力和杂种优势分析

对蜜蜂杂种F18个组合及其6个亲本的王浆产量和品质性状（台浆量、接受率、王浆产量、酸度）进行了配合力秘杂种优势分析。结果表明：⑴4个性状一般配合力达极显著水平、特殊配合力达显著水平,说明基因加性效应和非加性效应均起作用,但以加性效应为主。⑵乌克兰蜂一般配合力效应高;喀尔巴阡蜂、高加索蜂特殊配合力方差大,均是比较理想的亲本类型;浙农大1号意蜂在台浆量、接受率和王浆产量上一般配合力效应高,是个优良亲本;而卡尼鄂拉蜂只在提高王浆品质时可以利用。⑶4个性状高亲平均优势均为负值,因此利用杂种优质提高王浆产量和品质比较困难,但通过亲本的适当选配,可在维持高产的同时,提高王浆的品质。     

基于改进TOPSIS-灰色GM(1,1)模型的张家界市旅游生态安全动态预警

旅游生态安全预警对旅游地协调生态环境保护与旅游产业快速发展的矛盾,推进区域社会经济持续健康发展具有重要意义.本文基于DPSIR模型,从驱动力、压力、状态、影响、响应5方面构建了张家界市旅游生态安全预警指标体系,在此基础上,运用改进TOPSIS法对2001—2014年张家界市的旅游生态安全警情格局进行评估,并运用灰色GM(1,1)模型对其2015—2020年的警情演变趋势进行预测.结果表明: 总体上,2001—2014年,张家界市的旅游生态安全贴近度呈略微上升态势,警度处于“中警”.就各子系统而言,2001—2014年,张家界市旅游生态安全驱动力系统和压力系统的警度呈上升趋势,由“轻警”演变成“重警”;状态系统和影响系统的警度变化不大,一直处于“中警”;响应系统的警度呈下降趋势,由“巨警”转变成“无警”.按现状发展态势,2015—2020年,张家界市的旅游生态安全贴近度将进一步上升,警度将由“中警”向“轻警”转变,但协调旅游发展与生态建设和环境保护关系的任务依然较艰巨.     

农田向农林复合系统转变过程中土壤物理性质的变化

以渭北黄土区农林实践中被广泛采用的核桃-小麦间作复合模式为研究对象,以两物种的单作系统为对照,研究单作农田向农林复合系统转变对土壤物理性质的影响,为农林复合系统经营管理和模型的建立提供理论依据.结果表明: 核桃-小麦间作对土壤物理性质的改善作用主要发生在0～40 cm土层.核桃-小麦间作可以避免表层(0～20 cm)土壤容重升高,同时在20～40 cm土层对单作农田形成的犁底层也有显著的改善作用.核桃-小麦间作对各土层田间持水量均表现出持续的改善作用,除在20～40 cm土层略低于核桃单作外,其他从第5年开始均高于两单作系统.核桃-小麦间作对各土层土壤孔隙度均存在持续的改善作用,在0～20 和20～40 cm土层与两单作系统相比存在显著差异,同时也能提高毛管空隙度的比例.农田向农林复合系统转变过程中对土壤容重、田间持水量、土壤孔隙度均有持续的改善作用,且对浅层土壤的改善作用强于深层土壤.     

血红素加氧酶1在斑马鱼低氧应激中的保护作用研究

实验探讨了低氧条件下血红素加氧酶1（Ho1）对斑马鱼的保护作用。Real-time PCR结果显示,低氧条件下斑马鱼ho1 mRNA水平在斑马鱼胚胎和离体培养细胞ZF4中显著增加,而在成鱼的不同组织中呈现不同的反应。低氧处理24h后,斑马鱼脑、鳃和肝脏中ho1 mRNA表达量明显上升,而在心脏和肾脏中ho1 mRNA表达量显著降低。用锌原卟啉IX（ZnPPIX）抑制ZF4细胞ho1的表达,采用CCK8试剂盒检测细胞存活率,结果显示抑制ho1表达可导致低氧条件下ZF4细胞存活率明显降低。利用Hoechst染色和caspase 3活性检测发现,在低氧条件下抑制ho1表达后ZF4细胞的凋亡率较对照组显著增加,而Ho1的诱导剂可显著降低低氧条件下抑制组的细胞凋亡率。这些结果表明斑马鱼Ho1可能通过抗细胞凋亡发挥低氧保护作用。     

云南普洱季风常绿阔叶林非结构性碳水化合物及其组分随环境的变化

为探索植物非结构性碳水化合物(NSC)组分随环境的变化特征,在云南普洱季风常绿阔叶林原始林选取6块60m×60m样地进行野外调查,各样地分别选择重要值前10位的物种进行取样,通过测定不同样地主要物种的NSC及其组分和样地环境因素,分析其非结构性碳水化合物与环境的关系。结果显示:(1)云南普洱季风常绿阔叶林植物中总体淀粉、可溶性糖、NSC含量分别为13.91%、3.31%、17.21%,淀粉和NSC含量在各器官中排序依次为根干枝叶,而可溶性糖含量排序依次为叶根枝干;淀粉、可溶性糖、NSC含量变异最大值分别为叶片19.09%、干15.32%和根16.30%。(2)云南普洱季风常绿阔叶林植物不同器官中淀粉含量仅与凋落物厚度、土壤pH及速效钾、全钾存在显著正或负相关关系,可溶性糖含量与海拔高度及土壤理化性质存在显著正或负相关关系;但茶梨与红椎中淀粉含量与土壤有效磷或全磷存在显著正相关关系,茶梨各器官中可溶性糖含量与所有环境因子均无显著相关性,红椎根中可溶性糖含量与pH值、枝中可溶性糖含量与全钾分别呈显著负相关关系;短刺椎各器官中淀粉含量与所有环境因子均无显著相关性,可溶性糖含量则与水解性氮、有效磷含量呈显著负相关关系,与坡度、全钾含量、凋落物厚度呈显著正相关关系。(3)单因素回归分析显示,海拔高度、有机质、水解性氮、全氮影响季风常绿阔叶林的总体可溶性糖含量,而其淀粉含量则主要受pH值影响。研究发现,季风常绿阔叶林NSC受环境因子影响,但不同NSC组分的环境影响因子不同。