羊种布鲁氏菌M5-90疫苗株铁应答调控因子irr和rirA基因缺失株的构建与鉴定

为研究铁调控因子irr和rirA在羊种布鲁氏菌（Brucella melitensis）感染过程中的作用,本研究通过卡那替换的方法构建两个缺失株M5-90Δirr和M5-90ΔrirA,分别将亲本株和缺失株在相同营养条件下培养36 h,观察其振荡培养时的生长变化趋势。分别将1×10⁸ CFU M5-90Δirr、M5-90ΔrirA和M5-90接种到含1.5 mol/L NaCl、pH 2.5、pH 11.5、10 mmol/L H₂O₂的1 mL布鲁氏菌液体培养基,比较缺失株和亲本株在不同条件下的生长特性;分别以1×10⁶ CFU M5-90Δirr、M5-90ΔrirA和M5-90感染小鼠巨噬细胞RAW264.7检测缺失株的黏附、侵袭和胞内生存能力。结果显示,在相同体外培养条件下缺失株M5-90Δirr和M5-90ΔrirA生长速度明显低于亲本株;M5-90Δirr、M5-90ΔrirA在高盐、强酸和强碱环境下生存率均显著或极显著低于亲本株（P<0.05;P<0.01）,在H₂O₂条件下这两个缺失株的生存率却显著高于亲本株（P<0.05）。与亲本株相比,缺失株在小鼠巨噬细胞RAW264.7内的侵袭和黏附能力均减弱,在感染后45 min缺失株M5-90Δirr和M5-90ΔrirA的黏附和侵袭能力均极显著低于亲本株（P<0.01）,但在感染后24 h,这两个缺失株在细胞内繁殖能力与亲本株相比有所增强。本研究报道了irr和rirA基因不仅调控了羊种布鲁氏菌的生长,同时对细菌黏附和侵袭能力也产生一定的影响,M5-90Δirr和M5-90ΔrirA是两株具有潜力的布鲁氏菌候选疫苗株。     

两种木聚糖酶对黄羽肉鸡生长性能和养分代谢率的影响

试验旨在探讨不同来源木聚糖酶对黄羽肉鸡生长性能的影响。将1日龄健康黄羽肉鸡540羽,随机分成3组,每组6个重复,每重复30羽,试验期42 d。对照组饲喂小麦-玉米-豆粕型基础日粮,试验A、B组分别饲喂基础日粮中添加200 g/t木霉产木聚糖酶和200 g/t细菌产木聚糖酶的试验日粮,木聚糖酶酶活均为30 000 IU/g。结果表明,与对照组相比,试验A、B组的平均日增重和粗脂肪、粗纤维、钙、磷的表观代谢率及日粮表观代谢能均显著提高（P < 0.05）,料重比显著降低（P < 0.05）,粗蛋白质表观代谢率分别提高了5.90%（P < 0.05）和3.54%（P > 0.05）。试验A、B组之间各测定指标没有显著差异（P > 0.05） 。上述结果表明日粮中添加200 g/t木霉产木聚糖酶或细菌产木聚糖酶均能提高养分代谢率,改善肉鸡生长性能,木霉产木聚糖酶的效果稍好于细菌产木聚糖酶。     

The mechanism of apoptosis induced by homoharringtonine in HL-60 cells

AIM: To study the signal transduction pathway of apoptosis initiation induced by homoharringtonine in HL-60 cells. METHODS: After establishing the model of apoptosis initiation induced by homoharringtonine in HL-60 cells, at the point of apoptosis initiation, molecular caspase-3, Bcl-2, Bax and Fas/FasL were measured with flow cytometry and transmission electron microscope. ERK2 and P38 expression in HL-60 cells were detected by using immunohistochemistry. RESULTS: The model of apoptosis initiation induced by homoharringtonine was established in HL-60 cells. At the point of apoptosis initiation, upregulation of caspase-3 and decrease in Bcl-2/Bax were observed. However, the expression of Fas/FasL did not significantly change. ERK2 expression decreased and P38 expression increased. CONCLUSIONS: Caspase-3, Bcl-2, Bax and mitogen activated protein kinase pathways were involved in signal transduction of apoptosis initiation induced by homoharringtonine in HL-60 cells.     

基于信息熵的张掖市土地利用结构分析及其灰色预测

基于Shannon信息熵理论,分析了2003-2007年张掖市土地利用结构信息熵动态变化,对影响信息熵值大小的土地利用类型做了初步探讨,并运用灰色系统理论的GM(1,1)模型,对2008-2015年该市信息熵进行预测.结果表明:①影响张掖市土地利用结构信息熵动态演变的主要土地利用类型有牧草地、其他农用地、居民点及工矿用...     

出血性大肠杆菌O157eae-stx1/2B融合基因的构建和表达

构建表达eae和stx1／2B的融合基因，克隆eae基因的C端280个氨基酸残基(Int280)基因部分，以正确地阅读框定向插入到含有stx1／2B融合基因的质粒，构建重组质粒，将其转化于BL21(DE3)，用IPTG进行诱导表达，经SDS—PAGE电泳检测，该融合蛋白获得了高效表达。薄层扫描分析表明：目的蛋白表达量占菌体总蛋白含量的50．67％。由于该融合蛋白由eae、stx1B、stx2B等三部分抗原组成，可刺激机体产生针对紧密素和StxB的抗体，在EHEC O157亚单位疫苗设计或单克隆抗体抗制备中具有重要价值。     

团头鲂幼鱼饲料中α-亚麻酸、亚油酸的适宜含量

在半纯化饲料配方的基础上,分别设计6个α-亚麻酸含量(0.02%、0.55%、1.08%、1.60%、2.13%、2.65%)、6个亚油酸含量(0.86%、1.29%、1.73%、2.16%、2.59%、3.03%),以亚麻籽油、玉米油、棕榈油调节饲料中α-亚麻酸、亚油酸的含量,配制等氮等能(粗蛋白质含量为30.09%,粗脂肪含量为6.87%)的12种半纯化试验饲料,探讨团头鲂幼鱼[初始均重为(59.5±0.5)g]饲料中α-亚麻酸、亚油酸的适宜含量。养殖试验分为α-亚麻酸和亚油酸试验2部分,均设6组,每组4个重复,每个重复20尾,养殖周期为85 d。结果表明:在α-亚麻酸试验中,依据回归方程计算得到,在饲料α-亚麻酸含量分别为1.32%、1.33%时,团头鲂幼鱼具有最大的特定生长率和最小的饲料系数;0.02%组的脏体指数显著低于除1.08%组外的其他各组(P0.05),肥满度、肝体指数各组间无显著差异(P0.05);0.02%组血清总胆固醇、甘油三酯含量显著高于0.55%、1.08%组(P0.05),1.60%组血清高密度脂蛋白含量显著高于0.55%、2.13%组(P0.05),0.02%组血清低密度脂蛋白含量显著高于其他各组(P0.05)。在亚油酸试验中,依据回归方程计算得到,在饲料亚油酸含量分别为2.02%、2.03%时,团头鲂幼鱼具有最大的特定生长率和最小的饲料系数;3.03%组的肝体指数显著低于其他各组(P0.05),肥满度、脏体指数各组间无显著差异(P0.05);1.29%组血清总胆固醇含量显著高于1.73%组(P0.05),0.86%组血清甘油三酯含量显著高于1.73%组(P0.05),1.29%组血清高密度脂蛋白含量显著高于0.86%、3.03%组(P0.05),各组血清低密度脂蛋白含量无显著差异(P0.05)。以特定生长率、饲料系数作为主要评价指标,结合部分血清生化指标和形体指标,得到适合团头鲂幼鱼快速生长、维持鱼体正常健康的饲料中适宜的亚麻酸、亚油酸含量分别为1.32%~1.33%、2.02%~2.03%。     

工程菌DH5a感受态细胞的制备及质粒pGEM的转化研究

绿色荧光蛋白（green fluorescent protein，GFP）是源于海洋生物多管水母(aequoia victoria)的一种发光蛋白，能够自身催化形成生色团并在蓝光激发下发出绿色荧光，能在活细胞内稳定表达，本试验首先从含有GFP的大肠杆菌中提取质粒，并利用胶回收试剂盒纯化质粒，采用氯化钙法将工程菌DH_5a制成感受态细胞，然后将纯化后的质粒转入已制备好的感受态细胞中，使得工程菌DH_5a转化成具有抗氨苄青霉素的大肠杆菌，并表达绿色荧光蛋白，接种于加氨苄青霉素的LB培养基上，筛选阳性克隆。     

Effect of tetrandrine on radiosensitivity of nasopharyngeal carcinoma cells

AIM: To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS: The cells were treated with ma-ximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L), irradiation at 4 Gy, or combination of irradiation and maximum non-cytotoxic doses of Tet. The cell cycle distribution was analyzed by flow cytometry. The protein levels of γ-H2AX, cleaved caspase-3, p-CDC25C, CDK1, p-CDK1, cyclin B1, ERK and p-ERK were determined by Western blot.RESULTS: The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet. The percentages of CNE1 cells and CNE2 cells at G₂/M phase in irradiation group were (18.09±0.42)% and (18.48±1.32)%, respectively, which were decreased to (15.88±1.04)% and (13.80±0.82)% in combined treatment group, respectively (P<0.05). Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation. The protein levels of p-CDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P<0.05), while the expression of CDK1 showed no difference among different doses of Tet treatments. The protein levels of p-CDC25C, p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet. The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P<0.05), increased the expression of cyclin B1, and had no influence on the expression of CDK1 (P<0.05). The combined treatment resulted in an increase in the protein level of p-ERK1 (P<0.05).CONCLUSION: The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation, and the mechanism might be associated with ending of G₂/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.     

商丘市马铃薯“3414”肥效试验

马铃薯施用氮、磷、钾肥"3414"肥效试验表明,在商丘市睢阳区当地试验条件下,氮、磷、钾肥均影响马铃薯产量。通过回归分析,推荐春播马铃薯667 m2最佳经济施肥量为纯氮11.54 kg,纯磷2.26 kg,纯钾9.75 kg,667 m2产量达2 701.47 kg,最佳经济利润为3 109.46元。     

补播和划破草皮对退化亚高山草甸植被恢复的影响

在甘肃省甘南州临潭县研究围栏内补播和划破草皮对退化亚高山草甸植被的高度、总盖度、地上生物量和功能群物种多样性总指数的影响。结果表明,在各划破草皮处理中,牧草总盖度、地上生物量和功能群物种多样性总指数随着补播量的增加而增加,植被高度随着补播量的增加呈先增加后降低的趋势。补播和划破草皮有显著互作效果,补播量3[垂穗披碱草(Elymus nutans)播量为49.1 kg·hm-2,老芒麦(Elymus sibiricus) 播量为67.5 kg·hm-2,紫花苜蓿甘农1号(Medicago sativa cv.Gannong No.1)播量为23.3 kg·hm-2]和划破草皮组合的牧草地上生物量(201.5 g·m-2)、总盖度(88.2%)和功能群物种多样性总指数(1.203)达到最大值,补播量1（垂穗披碱草播量为26.7 kg·hm-2,老芒麦播量为37.5 kg·hm-2,紫花苜蓿甘农1号为0）和划破草皮组合的植被高度(11.7 cm)达到最大值。就各补播水平平均值而言,划破草皮植被高度、总盖度、地上生物量和功能群物种多样性总指数显著高于未划破草皮。综合考虑高度、总盖度、地上生物量和功能群物种多样性总指数,经关联度分析可知,补播量2（垂穗披碱草播量为37.5 kg·hm-2,老芒麦播量为52.5 kg·hm-2,紫花苜蓿甘农1号18.3 kg·hm-2）和划破草皮为最优组合。