鸡柔嫩艾美尔球虫病抗性主成分分析评估模型的建立

为建立鸡活体可测球虫病抗性综合评估模型,探索鸡球虫病抗性选育新方法,以京海黄鸡为试验材料,根据国内外文献选择15个球虫抗性评价指标,通过比较鸡柔嫩艾美尔球虫感染组和非感染组抗性指标的差异,进一步筛选活体可测指标,并进行主成分分析,通过各主成分和综合主成分与所有抗性指标的相关分析,筛选最优抗性综合评估模型,并进行模型有效性筛选。在所选的14个活体可测抗性指标中,有6个指标在感染和非感染组间存在显著或极显著的差异(P0.05或P0.01),他们分别是感染后第3—5天体增重、感染后第8天血浆中CAT、GSH-Px、MDA、SOD和IFN-γ的浓度;主成分分析建立了6个单主成分和累计贡献率大于80%的综合主成分的抗性评估模型,各抗性评估模型计算的综合选择指数与15个抗性指标相关分析的结果表明,第一主成分模型和7个抗性指标相关极显著(P0.01),2个相关显著(P0.05),特别是与盲肠病变记分相关极显著(P0.01),其余6个主成分评估模型只与15个抗性指标中的2~3个相关显著或极显著(P0.05或P0.01),且与盲肠病变记分相关均不显著(P0.05);经检验第一主成分模型选择指数值的分布为正偏态分布,排序后其大小与盲肠病变表现一致。因此,第一主成分模型fi1=-0.64Zxi1+0.31Zxi2+0.80Zxi3-0.05Zxi4-0.08Zxi5+0.59Zxi6可以作为鸡柔嫩艾美尔球虫抗性选择的最佳综合选择指数用于鸡的抗病育种。     

First report of Pseudomonas aeruginosa causing tumor disease of Populus koreana in China

Journal of Plant Diseases and Protection - In May 2018, a bacterial tumor disease was observed on Populus koreana in Shuangyang county, Changchun city, Jilin Province. Based on physiological...     

干旱胁迫下硫对小麦叶片GSH含量及GSH-PX活性的影响

生物体内的谷胱甘肽对消除逆境产生的活性氧具有重要的作用.无机硫营养吸收与体内有机硫的转化有着密切相关,研究干旱胁迫下硫营养与谷胱甘肽(GSH)含量及谷胱甘肽过氧化酶(GSH-PX)活性的关系,对揭示硫素的抗逆作用机理具有重要的理论和实际意义.本研究以高水肥型小麦品种郑引1号和抗旱型品种陕合6号为供试材料,分别用Hoagland全营养液和缺硫营养液进行溶液培养试验,探讨了模拟干旱胁迫过程下小麦苗期功能叶片中GSH含量和GSH-PX活性的变化.结果表明:陕合6号的GSH含量显著高于郑引1号;水分胁迫下,郑引1号中供硫处理的GSH含量上升迅速,24 h、48 h及72 h分别比供水处理增加了26.4%、33.5%和56.6%,而缺硫处理则分别是供水处理的93.2%、95.9%和74.5%;陕合6号在水分胁迫的72 h内供硫处理的GSH含量变化不明显,分别为供水处理98.3%～106.1%,而缺硫处理的GSH含量有明显的增加,分别比供水处理增加了13.1%、40.2%和45.9%.郑引1号的GSH含量受水分胁迫和硫营养供给状况的影响显著,而陕合6号的GSH含量仅在缺硫情况下对水分胁迫敏感.水分胁迫初期(24 h)可诱导GSH-PX活性增高,郑引1号供硫处理较供水处理增加了23.9%,而缺硫处理仅增加了7.4%;陕合6号品种的供硫处理和缺硫处理分别较供水处理增加了24.5%和16.8%.随着干旱胁迫时间的延长(48～72 h),GSH-PX活性均呈降低趋势,但供硫处理的GSH-PX活性始终高于缺硫处理.郑引1号高水肥型品种的GSH-PX活性较低,在缺硫环境下受水分胁迫的诱导不敏感;而陕合6号抗旱型品种的GSH-PX活性整体上较高,并且对水分胁迫的响应比较敏感.     

应用SYBR Green I的Real-time PCR方法定量检测空肠弯曲杆菌

空肠弯曲杆菌(Campylobacterjejuni)被认为是人类主要食源性病原菌之一。由于空肠弯曲杆菌特殊的生长条件和容易进入不可培养但存活的状态(Viablebutnonculturable,VNC),所以传统的生化鉴定结果不一定可靠,并且是一项费时而繁琐的工作。核酸检测方法的出现为空肠弯曲杆菌的检测带来了方便。本文基于LightCycler为平台,利用SYBRGreenI染料能特异与双链DNA结合而发荧光的特性,建立一种Real-timePCR方法来定量检测空肠弯曲杆菌。用该方法检测时,发现所有空肠弯曲杆菌(11株)都呈阳性,所有其它弯曲菌(3株)和其它菌株(5株)都是阴性。整个检测过程60min内可以完成,检测限度为5CFU,标准曲线的相关系数为1。结果表明基于SYBRGreenI的定量PCR方法既为空肠弯曲杆菌提供了一种特异、敏感、快速和简洁的定量检测方法,又为研究空肠弯曲杆菌致病机理提供了一种重要方法。     

重寄生真菌盾壳霉胞外蛋白酶产生条件及酶活影响因子

重寄生真菌盾壳霉Coniothyrium minitans是核盘菌Sclerotinia sclerotiorum的重要生防菌。为了探讨盾壳霉胞外蛋白酶在寄生核盘菌过程中的作用,采用明胶平板法对盾壳霉寄生核盘菌菌核产生的蛋白酶活性进行了检测,并进一步采用福林酚法定量测定蛋白酶活性,研究盾壳霉产生胞外蛋白酶的培养条件及影响蛋白酶活性的因子。试验结果表明,在被盾壳霉寄生的核盘菌菌核中检测到蛋白酶活性,表明蛋白酶可能参与盾壳霉重寄生作用。发现核盘菌菌核浸出液培养基适合盾壳霉产生胞外蛋白酶,摇培(20℃、200r/min)5d时蛋白酶活性最高,达到0.22U/mL。盾壳霉胞外蛋白酶酶促反应的最适温度为60℃,最适pH7.0。当温度不高于40℃时,蛋白酶酶活较稳定。5mmol/L的金属离子Mg²⁺、Zn²⁺、Ca²⁺、Cu²⁺、Mn²⁺、Li⁺和K⁺等对蛋白酶酶活没有显著影响(P>0.05),而Fe²⁺(5mmol/L)显著(P<0.05)提高了蛋白酶活性。盾壳霉蛋白酶对苯甲基磺酰氟(PMSF)敏感,说明盾壳霉产生的胞外蛋白酶可能主要是丝氨酸蛋白酶。这些结果为盾壳霉胞外蛋白酶的分离纯化和功能研究奠定了基础。     

Effect of exogenous hydrogen sulfide on lipophagy in mouse primary hepatocytes

AIM: To evaluate the effect of exogenous hydrogen sulfide (H₂S) from GYY4137 on lipophagy in mouse primary hepatocytes. METHODS: The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups: the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h; the cells in H₂S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope, phase-contrast microscope or transmission electron microscope. The protein expression of LC3-Ⅰ/Ⅱ in the hepatocytes was determined by Western blot. RESULTS: In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H₂S group increased. CONCLUSION: In steatosis hepatocytes, exogenous H₂S promotes the lipophagy.     

禽呼肠孤病毒分子生物学研究进展

本文就十几年来有关呼肠孤病毒的特性、基因组、基因重组、病毒蛋白、诊断方法等分子生物学的研究进行了综述,并指出了禽呼肠孤病毒分子生物学研究中存在的问题,及进一步研究的方向。     

基于GIS与马尔可夫模型的渭河盆地景观动态变化研究

基于1988年与2003年两期TM影像,综合遥感与GIS技术以及景观生态学理论,深入分析了渭河盆地景观变化的数量和空间特征以及由此所引起的一系列生态环境效应,主要表现在:耕地、园地与城镇建设用地面积显著增加,林地、水体和未利用地面积呈减少趋势,城镇建设用地的增加是以侵占大量农田为代价的,而园地与耕地的增加则主要是通过开荒的方式。景观形状趋于复杂,破碎度降低,异质性下降,景观类型有向单一化或非均衡化方向发展的趋势,表明人类的开发活动增强,这样必然导致景观稳定性下降,因而降低了景观抗干扰的能力。最后采用马尔可夫模型对景观的变化趋势进行了预测。     

Radioligand binding assay of insulin receptor in rabbit kidney during ischemic and reperfusion

AIM: To observe the change of insulin receptor in rabbit kidney with acute ischemic-reperfusion injury. METHODS: 15 Japanese white rabbits were allocated randomly into control group, ischemic-reperfusion group(IR group). IR group received clamping for 1 h followed by 2 h or 48 h of reperfusion. At 2 h or 48 h after reperfusion, glucose and insulin in serum were determined. Insulin receptor in renal tissue was analyzed by radioligand binging assay(BAD). RESULTS: The level of serum glucose increased after 2 h reperfusion in 2 groups, but in IR group the value increased much more higher than those in control groups(P＜0.05). Plasma insulin of IR group was significantly higher than that in control after 2 h reperfusion(P＜0.05). Scatchard analysis of data resulted in curvilinear profiles, indicating that there are two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed that the values of Bmax₁(high affinity site), Bmax₂(low affinity site) and Kd₁, Kd₂ were significantly lower than that of control (P＜0.05) after 2 h perfusion. 48 h after IR there was no difference of Bmax₁, Bmax₂, and Kd₁ between 2 groups,but Kd₂ of IR group was higher than that of control (P＜0.05). CONCLUSIONS: The results indicate that the effect of intrinsic insulin decreases in the progress of the renal ischemic-reperfusion. The resulting high serum glucose may aggravate renal injury in the progress of ischemic-reperfusion.     

Suberoylanilide hydroxamic acid induces apoptosis of HepG2 cells by endoplasmic reticulum stress apoptotic pathway

AIM: To investigate the effect of suberoylanilide hydroxamic acid (SAHA) on the proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells and to explore its possible mechanism. METHODS: HepG2 cells were treated with SAHA at different concentrations for 48 h. The proliferation of HepG2 cells was detected by real-time cellular analysis. The protein levels of acetylated histones H3K9 and H3K27, glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase (PERK) and p-PERK were determined by Western blot. The cell apoptosis was analyzed by flow cytometry. RESULTS: Compared with control group, treatment with SAHA at 0.1 μmol/L and 1 μmol/L for 48 h showed no significant inhibitory effect on the proliferation of HepG2 cells, while SAHA at 6 μmol/L and 12 μmol/L significantly inhibited the proliferation of HepG2 cells (P<0.05). The results of Western blot showed that the protein levels of acH3K9, acH3K27, GRP78 and p-PERK increased significantly after treated with SAHA at diffe-rent concentrations for 48 h, while the protein level of PERK was decreased significantly (P<0.05). The results of flow cytometry analysis showed that the apoptotic rates of the HepG2 cells increased with the increase in SAHA concentration. CONCLUSION: SAHA up-regulates the acetylation of H3K9 and H3K27 in the HepG2 cells and induces apoptosis of HepG2 cells by activating the endoplasmic reticulum stress-related apoptosis pathway.