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451.
白菜亚硝酸还原酶基因BcNiR的克隆及表达分析 总被引:2,自引:0,他引:2
以白菜品种‘苏州青’自交系叶片cDNA为模板, 采用RT-PCR、3′RACE和5′RACE技术,
获得了编码亚硝酸还原酶基因(NiR) 的cDNA全序列1 852 bp, 包含有1 749 bp的开放阅读框, 编码583个氨基酸, 命名为BcNiR。所推导的氨基酸序列与拟南芥NiR1、烟草nii2编码的氨基酸序列具有较高同源性, 分别为83%和76%。生物信息学分析表明, BcNiR具有完整的NiR蛋白结构, 含血红素蛋白β - 化合物区域, 一个明显的西罗血红素siroheme结合位点和4Fe-4S区域, 可以在SWiSS-MODEL数据库中搜索到与之相近的三维结构。RT-PCR结果显示, 该基因在叶片中的表达量远远高于根系, 在以0、10、20、30、40 mmol·L - 1硝态氮各分别处理0、2、4、6、8、12 h的试验中, 30 mmol·L -1处理4 h可使BcNiR的表达量达到最大; 5和10 mmol·L - 1铵态氮处理试验表明, 高浓度的铵抑制该基因的表达。 相似文献
452.
‘岩溪晚芦’椪柑果皮与果肉差减cDNA文库的构建及初步分析 总被引:4,自引:0,他引:4
以‘岩溪晚芦’(Citrus reticulata Blanco)成熟果皮和果肉为材料,采用抑制性差减杂交技术,成功构建了果皮(tester)与果肉(driver)的差减cDNA文库。结果显示:cDNA克隆外源插入片段大小介于100~500 bp之间。成功对411个克隆进行了测序,测序结果经与NCBI基因库中序列进行比较,377个EST 找到了同源序列,它们分属于126个基因,涉及到与色素合成、能量代谢、初级代谢、次生代谢、抗逆防御、蛋白代谢、信号转导、香味形成、果皮软化(衰老)、精油合成、抗氧化代谢等代谢途径和生理生化过程。还有一些基因涉及到三价铁螯合物、反转录转座子gag蛋白、查尔酮合成、硼离子转运、儿茶酚氧位甲基转移酶等功能。功能已知且没有重复的EST共有63个,占总数的49.2%。有5个基因共62个克隆虽在Blast搜索时找到了较高的同源性序列,但功能目前尚未明确,有待进一步深入分析。另有34个基因未搜索到同源序列,可能为新发现的基因。 相似文献
453.
AIM: To observe the effects of carvedilol on murine viral myocarditis model. METHODS: A total of 188 inbred male BALB/c mice of 4-6 weeks were divided into 4 groups: myocarditis group (group C, n=60), metoprolol treatment group (group M, n=60), carvedilol treatment group (group K, n=60), control group (group B, n=8). Myocardial histopathololgic changes were observed. The concentrations of cardiac troponin I (cTn-I) were detected by chemiluminescence immunoassay (CLIA). Western blotting was performed to analyze the contents of phosphorylated p38MAPK in myocardium. RESULTS: Metropolol and carvedilol lightened myocardial histopathololgic changes at acute stage, decreased cTn-I concentrations and myocardial phosphorylated p38MAPK value compared with myocarditis group. Treatment with carvedilol was more effective than treated with metropolol on those indexes. CONCLUSION: Carvedilol protects against viral myocarditis by inhibition of p38MAPK signal transduction pathway through blockade of β1 and β2 adrenergic receptors. 相似文献
454.
AIM: To observe the damage induced in the primary cultured rat cortical neurons by oxygen/glucose deprivation and reintroduction, and to investigate the neuroprotective mechanism of salvianolic acid B (SalB). METHODS: Primary cultured rat cortical neurons were randomly divided into the control group, the model group and the SalB group. The cell model was established by oxygen/glucose deprivation for 3 h followed oxygen/glucose reintroduction for 24 h. The cortical neurons viability was determined by MTT assay. The leakage rate of lactate dehydrogenase (LDH) was measured by chromatometry. The mitochondrial membrane potentials (MMP) and the apoptosis rate were quantitatively analyzed by flow cytometry. The cytosolic free calcium was assessed using LSCM. The morphologic changes of neuronal nuclei were observed by Hoechst 33342 fluorescence staining. RESULTS: Compared to the model group, the cortical neurons viability, the survival rate and the fluorescence value of MMP in the SalB group were obviously increased (P<0.05,P<0.01). In addition, in the SalB group, the leakage rate of LDH, the fluorescence intensity of cytosolic free calcium and the apoptosis rate were significantly lower than those in the model group (P<0.01). CONCLUSION: The neuroprotective mechanism of SalB in the oxygen/glucose deprivation and reintroduction neurons would be due to the fact that SalB maintains the MMP and the calcium homeostasis. 相似文献
455.
456.
JIANG Wen-qiang FANG Ming CHEN Chun-bo WANG Qiao-sheng YE Heng ZENG Hong-ke 《园艺学报》2009,25(4):755-758
AIM: To investigate the role of stress hyperglycemia on condition assessment and predicting prognosis in patients with sepsis. METHODS: The study included 44 patients with sepsis, divided into three groups according to their blood glucose profile within 24 h after admission: patients with stress hyperglycemia (group SH, n=15), diabetes mellitus type 2 (group DM, n=10), and normal glucose levels (group NG, n=19). CD4+/CD8+ ratio, Th1/Th2 ratio and HLA-DR% of the patients were measured within 24 h after admission by flow cytometry as assessment of their immune function. The sepsis-related organ failure assessment (SOFA) scores and acute physiology and chronic health evaluation II (APACHE II) scores of patients were recorded at the same time to analyze whether stress hyperglycemia affects the immune function and the 28 d mortality in patients with sepsis. RESULTS: A higher mortality rate of septic patients with stress hyperglycemia was observed compared to diabetic patients (53.3% vs 10.0%) and group NG (53.3% vs 21.1%). SOFA score and APACHE II score were higher in group SH than those in group DM and group NG while lower levels of human leucocyte antigen DR (HLA-DR) expression and CD4+/CD8+ ratio was found in group SH than those in group DM and group NG. No difference in the levels of Th1/Th2 among the three groups was observed. Non-survivors had higher levels of SOFA score, APACHEⅡ score, Th1/Th2 ratio and HLA-DR%. No difference was detected for CD4+/CD8+ ratio, mean glucose values and age. CONCLUSION: Stress hyperglycemia is associated with decreased immune function and an adverse clinical outcome in patients with sepsis. 相似文献
457.
ZHENG Ming-zhi JIANG Jian-ping CHEN Wen-liang ZHANG Xiao-bing ZHU Li SHEN Yue-liang CHEN Ying-ying 《园艺学报》2009,25(9):1676-1680
AIM: To investigate the effect of a mitochondrial ATP-sensitive potassium channel (mitoKATP) opener diazoxide (DE) on Smac/DIABLO protein expressions in rat heart suffered from different duration of hypothermic preservation. METHODS: The Langendorff model of isolated rat heart was used. After stored in 4 ℃ Celsior solution with or without DE (30 μmol/L) for different time (0, 3, 6, 9 or 12 h). Cell apoptosis was detected by TUNEL technique. The expression of Smac/DIABLO protein in cytoplasm and total caspase-3 protein in myocardia tissue was also analyzed by Western blotting. RESULTS: (1) Compared to the hypothermic preservation groups, DE reduced the percentage of apoptotic cells and the expression of caspase-3 protein in myocardia tissue. (2) The peak of Smac/DIABLO protein expression level appeared at 6 h after hypothermic preservation, and which was postponed to 9 h by DE. (3) The above effects of DE were attenuated by a mitoKATP channel inhibitor 5-hydroxydecanoate (5-HD). CONCLUSION: The findings indicate that in the isolated rat heart, DE protects myocardium against different duration of hypothermic preservation injury via opening of mitoKATP channel and inhibition of Smac/DIABLO protein expression. 相似文献
458.
AIM: To investigate the mechanisms of transdifferentiation of renal tubular epithelial cells by the observation of serial activation of embryonic pax2 and WT1 genes in chronic renal failure model of 5/6 nephrectomized injury. METHODS: Embryo of mice, cultured renal tubule cells and chronic renal failure rat model of 5/6 nephrectomized injury were established. The expressions of paired Box gene (pax2) and Willm’s tumor gene (WT1), as well as α-smooth muscle actin (α-SMA), a phenotype of mesenchymal cells, were detected by RT-PCR and immunohistochemistry techniques. RESULTS: (1) Expressions of pax2 and WT1 mRNA began at 11.5 d and 14 d of embryo and increased gradually, and expression in trace quantity at 2 weeks after birth. Immunohistochemistry analysis revealed that WT1 only expressed in the glomerular podocytes and expressions of pax2 and WT1 in adult renal tubular cells were negative. (2) Deno-expression of pax2 and WT1 in some tubular cells appeared at week 2 and peaked at week 4 in 5/6 nephrectomized rats, both showing a same trend of expression. However, pax2 showed another peak at week 10 afterwards. (3) Re-expressions of pax2 and WT1 in TEC at 0.5 and 24 h after treated with IL-1α (10 μg/L) or AngII (10-9 mol/L) were observed respectively, followed by upregulation of α-SMA expression, and mesenchymal cells characters were shown. The effects were inhibited by Ang II receptor 2 antagonist and WT1 antibody, respectively. CONCLUSION: Adult renal tubule cells acquire re-activation of embryonic pax2 and WT1 genes and phenotypes of mesenchymes when challenge with certain injuries. Deno-expression of pax2 and WT1 genes closely relates with high concentration of bioactive facors, such as AngII and IL-1, which are involved in the mechanisms of renal tubule cells transdifferentiation. 相似文献
459.
460.