A Botrytis cinerea Putative 3-keto Reductase Gene (ERG27) that is Homologous to the Mammalian 17β-Hydroxysteroid Dehydrogenase type 7 gene (17β-HSD7)

Botrytis cinerea (anamorph of Botryotinia fuckeliana) is a filamentous ascomycete that causes grey mould on grapevine. We had previously described two distinct populations, named HydR1 and non-HydR1, that comprise two distinct genetic entities based on genetic polymorphism, natural resistance towards the fungicide hydroxyanilide fenhexamid, and vegetative incompatibility between them. Here, we used PCR to isolate the 3-keto reductase gene ERG27 by virtue of sequence homology with Saccharomyces cerevisiae ERG27. The gene product was longer than the yeast's enzyme but possessed the main characteristic features of reductases. It displayed striking homology with mammalian 17-HSD7, therefore confirming the hypothesis of a common function between Erg27p like protein and 17-HSD7 in sterol biosynthesis (i.e. cholesterol, ergosterol). On the other hand, we analysed the polymorphism of the B. cinerea gene product and found a dozen of amino-acid differences between strains of HydR1 and non-HydR1 types that could underlie HydR1 natural resistance to fenhexamid. First, this polymorphism analysis showed that HydR1 strains form a homogeneous group distinct from the non-HydR1 group of strains. These results support our hypothesis that HydR1 and non-HydR1 strains constitute two different species. Second, Erg27p like protein sequence analysis showed that a high resistant phenotype to fenhexamid, HydR3, found in treated populations of non-HydR1 strains, had two mutations (usually found in mammalian 17-HSD7) that could be useful as population markers.     

Role of p38 MAPK-HSP27 signaling pathway in rat acute lung injury induced by lipopolysaccharide

AIM: To observe the role of p38 mitogen-activated protein kinase (p38 MAPK)-heat-shock protein 27 (HSP27) signaling pathway in lipopolysaccharide-induced acute lung injury (ALI) in rats. METHODS: Wistar rats were randomly divided into control group, ALI group and ALI+SB203580 group. After the experimental model was established, the rats were sacrificed. The pathological changes of the lung and the changes of F-actin and G-actin in the endothelial cells were observed. The ratio of wet weight to dry weight (W/D) of the lung tissues was measured. The protein levels in bronchoalveolar lavage fluid (BALF) were detected. The levels of IL-6 and TNF-α in serum and BALF were tested. The concentrations of p-p38 and p-HSP27 in the lung were determined. RESULTS: In ALI group, the protein levels in BALF and W/D ratio of the lung increased significantly at 2 h. The levels of TNF-α and IL-6 in serum and BALF began to increase at 2 h, which had significant difference as compared with control group. Aleolar epithelial swelling, alveolar walls widening, alveolar interstitial and cavity edema, and the exudation of alveolar inflammation cells, red blood cells and protein were observed in ALI group. The protein levels in BALF and W/D ratio of the lung in ALI+SB203580 group were much less than those in ALI group. The exudation of alveolar inflammation cells, red blood cells and protein, and the interstitial and alveolar edema in ALI+SB203580 group alleviated as compared with ALI group. The expression of p-p38 MAPK and p-HSP27 in the lung at 2 h in ALI group was higher than that in control group. F-actin expression in ALI group obviously increased than that in control group at time points of 0 h and 8 h. Compared with ALI group, the expression of p-HSP27 and F-actin in ALI+SB203580 group was reduced. CONCLUSION: Lipopolysaccharide activates p38 MAPK-HSP27 signaling pathway and induces lung injury. Blockage of p38 MAPK-HSP27 signaling pathway may reduce lung injury.     

禽白血病病毒抗体间接ELISA 检测方法的建立及初步应用

以大肠杆菌表达、纯化的禽白血病病毒（ALV）重组P27蛋白为包被抗原,建立了检测ALV-P27抗体的间接ELISA方法,为禽白血病流行病学调查提供了一种简便的血清学检测方法.该方法与9种禽常见传染病病毒阳性血清及大肠杆菌阳性血清无交叉反应,具有较好的特异性;批内、批间重复试验变异系数均小于10％,具有良好的重复性;新建立的ELISA方法比IDEXX公司生产的ALVA、B亚群抗体检测试剂盒和J亚群抗体检测试剂盒相对于免疫荧光试验（IFA）有更高的符合率.     

不同物种RAB27A基因完整编码区生物信息学分析

应用比较基因组学和生物信息学方法比较分析了人、黑猩猩、恒河猴、家鼠、挪威鼠、狗、猪、牛、马、短尾负鼠、鸡和非洲爪蛙RAB27A基因完全编码区(cds),并对该基因的遗传多样性及分子系统发育进行了分析。结果表明,来自12个物种的52条基因序列检测到239个多态位点,共生成17种单倍型。RAB27A基因物种内保守性较强,但在物种间核苷酸多样性和氨基酸多样性相对丰富。     

Skp2与血管平滑肌细胞增殖

血管平滑肌细胞(VSMC)增殖的过程涉及到细胞周期的调控,S期激酶相关蛋白2(Skp2)能介导泛素化蛋白通过泛素蛋白酶降解途径进行降解,参与了细胞周期调控,与VSMC增殖密切相关.本文就Skp2与VSMC的增殖关系作一综述.     

绵羊Fsp27基因组织表达、多态性及其与不同绵羊品种尾脂沉积能力的关联性分析

旨在研究绵羊Fsp27基因的组织表达/多态性及其与不同绵羊品种尾脂沉积能力的关联性。本研究采用qRT-PCR检测了营养充足期阿勒泰羊不同组织中Fsp27基因的表达情况,同时对营养充足期和营养匮乏期阿勒泰羊尾脂组织中Fsp27基因的表达情况进行了定量分析,采用PCR-SSCP结合测序技术检测了5个不同尾脂沉积能力绵羊品种Fsp27基因的突变情况,并分析了相关突变与绵羊尾脂沉积能力的关联性。结果表明,Fsp27基因在营养充足期阿勒泰羊尾脂中高表达,其表达量极显著高于其他组织（P<0.01）,在肾周脂肪和皮下脂肪组织中的表达量也较高,极显著高于心、肝、脾、肺、肾、胃、肠、皮肤和骨骼肌组织（P<0.01）。在心、肝、脾、肺、肾、胃、肠、皮肤和骨骼肌组织中呈微量表达,且各组织间差异不显著（P>0.05）。另外,营养充足期阿勒泰羊尾脂中Fsp27基因的表达量极显著高于营养匮乏期（P<0.01）。绵羊Fsp27基因在不同尾脂沉积能力品种群体中共检测到25个突变位点,其中位于第3外显子g.16771741的G/A突变以及第5外显子g.16774969的C/T突变均为错义突变,且与绵羊尾脂沉积能力高度相关。g.16771741的G/A突变在尾脂沉积能力较强的阿勒泰羊、小尾寒羊和湖羊群体中以G等位基因为主,分别占到86.8%、83.7%和85.7%,而尾脂沉积能力较差的中国美利奴细毛羊和萨福克羊群体中G等位基因分别仅占30.3%和11.1%。g.16774969的C/T突变在阿勒泰羊群体中以T等位基因为主,TT基因型占84.7%,CT基因型占15.3%,没有检测到CC基因型;在短脂尾型的小尾寒羊和湖羊群体中,TT和CT基因型也分别占到65.9%、50.4%和22.7%、41.1%,而在长瘦尾的中国美利奴细毛羊和萨福克羊群体中则以CC基因型为主,分别占到97.4%和69.4%,没有检测到TT基因型。以上研究结果表明,Fsp27基因在阿勒泰羊尾脂中高表达,且其在尾脂中的表达量与阿勒泰羊的营养状态密切相关,Fsp27基因第3外显子g.16771741的G/A突变以及第5外显子g.16774969的C/T突变与绵羊尾脂沉积能力高度相关,可以作为理想的分子标记用于低脂肪绵羊品种的选育。     

SC27菌剂对黄瓜、西红柿、甜椒生长代谢、产量和品质的影响

田间试验结果表明 ,土壤浇施 SC2 7菌剂 3.0、6.0 L· hm-2 均可提高黄瓜、西红柿、甜椒产量 1.5- 6.7t· hm-2 ,增幅 4 .17% - 14.63% .这与 SC2 7促进土壤养分有效化 ,提高植株叶片中 N、可溶性蛋白质、糖和淀粉含量有关 .试验同时发现 ,SC2 7可以增加黄瓜、西红柿和甜椒中 17种氨基酸含量 ,尤其是人体必需氨基酸含量 ,以及果实中蛋白质、糖、Ca、P、Fe、Zn和 VC含量 ,改善果实品质     

Effects of HIF-1α silencing on expression of p27 and Ki67 in rat hepatoma cell line CBRH-7919

AIM: To study the effect of hypoxia-inducible factor 1α(HIF-1α) silencing on the expression of p27 and Ki67 in rat hepatoma cell line CBRH-7919 under hypoxia. METHODS: Hypoxic condition was induced by CoCl₂ and the expression of HIF-1α was silenced by small interference RNA. HIF-1α-specific RNAi lentiviral vector was constructed. Real-time RT-PCR and Western blotting were used to detect the expression of HIF-1α at mRNA and protein levels in CBRH-7919 cells under hypoxia. The expression of p27 and Ki67 was observed by Western blotting after HIF-1α silencing was performed. The cell cycle of hepatoma cells was detected by flow cytometry. RESULTS: Under hypoxic condition, the expression of HIF-1α at mRNA and protein levels in CBRH-7919 cells increased significantly (P<0.05). HIF-1α silencing significantly reduced the expression of Ki67 but increased the expression of p27 (P<0.05) in CBRH-7919 cells. In transfected cells, the number of cells in G₀/G₁ phase was much higher and that in S phase was much lower than those in the control cells. CONCLUSION: Hypoxia induces the expression of HIF-1α. HIF-1α silencing can regulate the proliferation of hepatoma cells through reducing the expression of Ki67 and increasing the expression of p27.     

沙门氏菌和聚肌胞处理条件下鸡免疫相关候选基因mRNA表达特性

【目的】利用高密度SNP芯片完成了对北京油鸡血清免疫球蛋白Y含量等9个免疫性状的关联分析,筛选得到了与性状显著关联位点和候选基因。基于该研究结果,以集中分布在16号染色体的候选基因CD1b、 BMA1（B locus M alpha chain 1）、TRIM27（tripartite motif-containing 27）和ZNF692（zinc finger protein 692）作为研究对象,以北京油鸡为实验材料,在聚肌胞（Polyinosinic acid-polycytidylic acid, Poly I:C）和细菌的处理下进一步对候选基因表达特性进行分析和鉴定。【方法】随机选取80只12日龄北京油鸡分为3组：空白对照组、聚肌胞处理组和肠炎沙门氏菌（Salmonella enteritidis, SE）处理组,分别饲养在独立的隔离器内。两处理组分别胸肌注射聚肌胞注射液和SE菌液,对照组注射生理盐水,于处理后12 h、24 h、3 d和6 d（days post infection, DPI）检测血清炎症因子的变化水平和候选基因表达规律。【结果】经过聚肌胞和SE处理后,体重在24 h之后显著低于空白组（P＜0.05）,体温24 h以内显著升高;血清IFN-α、IL-4和IL-6水平先升高后降低,在第24小时或第3天达到峰值,TNF-α水平持续升高,3 d后极显著高于空白组（P＜0.01）。两种处理条件下CD1b的mRNA表达没有组织特异性,其他3个基因在胸腺和法氏囊中高表达。在胸腺组织中,CD1b在两处理间12和24 h的表达量存在显著差异（P＜0.01）,在聚肌胞处理组整个感染阶段没有显著性变化,在SE组是先升高后降低的趋势,感染后24 h时表达量最高（P＜0.01）;BMA1在12 h和3 d时两处理组间差异显著,聚肌胞处理组在12 h时表达量低于SE处理组,而在3 d时显著高于SE处理组（P＜0.01）;TRIM27在聚肌胞感染6 d时表达量显著高于空白组（P＜0.05）,ZNF692的表达量在3组间没有差异。在法氏囊中,CD1b表达量在两个处理组中存在差异,在SE组12 h时表达量高;BMA1和TRIM27的表达量在两组间没有差异,TRIM27在3 d时表达量最高（P＜0.01）,ZNF692在SE组感染24 h时相对表达量高于聚肌胞处理组,在两组中都在3 d时表达最高。【结论】CD1b、BMA1、TRIM27和ZNF692参与了聚肌胞和肠炎沙门氏菌引起的免疫反应过程,是与免疫相关的功能基因;CD1b主要在肠炎沙门氏菌感染的前期发挥作用;BMA1和ZNF692分别参与胸腺和法氏囊的免疫反应。     

SC27菌剂对芦柑产量和品质的影响

多点田间试验结果表明 ,每年土壤兑水浇施 60 m L·株 - 1 澳州 SC2 7菌剂可显著提高芦柑产量 ,增产幅度达 4.2 2 % -6.80 % ,这与 SC2 7菌剂可促进土壤养分有效化 ,提高植株叶片 N、蛋白质、糖和淀粉含量有关 .SC2 7菌剂还可增加芦柑中1 7种氨基酸含量 ,尤其是人体必需氨基酸含量 ,以及果实中蛋白质、糖、可溶性固形物、VC、P、Ca、Fe、Zn等含量 ,改善果品品质