竹荚鱼属鱼类线粒体DNA控制区结构及其系统发育分析

采用PCR产物直接测序法测定了2尾来自中国南海的日本竹煲鱼（Trachurus japonicus）线粒体DNA控制区基因全序列,并结合从GenBank中下载的8种竹笑鱼属相应序列,对竹煲鱼属控制区序列进行了结构分析,识别出终止序列区、中央区和保守序列区3个区域,并找到了2个与DNA复制终止相关的序列TAS和中央保守区的保守序列CSB—F、CSB—E和CSB—D,以及保守序列区的保守序列CSBl、CSB2和CSB3。以鳜（Sinipercachuatsi）作为外群,使用邻接法和最大简约法构建了9种竹荚鱼属的系统发育树。竹笑鱼属鱼类为单系类群,蓝竹煲鱼（rpicturatus）、智利竹笑鱼（zmurphyi）、太平洋竹笑鱼（zsymmetricus）、南非竹荚鱼（rcapensis）和竹荚鱼（rtrachurus）构成一支,地中海竹笑鱼（zmediterraneus）、青背竹笑鱼（zdeclivis）、日本竹荚鱼（rjaponicus）和新西兰竹荚鱼（rnovaezelandiae）为另一支;日本竹笑鱼未单独成一支,而是聚人青背竹笑鱼和新西兰竹笑鱼之间,初步猜测日本竹笑鱼和新西兰竹荚鱼为同种异名。     

猪-鱼复合养殖模式中气单胞菌I类整合子的流行情况及其耐药特征

为了解广东地区猪-鱼复合养殖模式下气单胞菌整合子流行情况及其耐药特征,从广东省5个不同猪-鱼复合养殖场采集分离猪粪、鱼、池塘水及池塘底泥的气单胞菌共317株,通过微量二倍稀释法测定其对20种药物的敏感性;PCR扩增Ⅰ类整合子整合酶基因intI1,并分析其基因盒阵列结构。结果表明,317株气单胞菌对20种药物耐药程度不一,氨苄西林、磺胺间甲氧嘧啶、萘啶酸的耐药率相对较高。intI1的检出率为15.77%,鱼源和猪源的整合子检出率高于环境源。整合子阳性菌对测试的12种药物的耐药率显著高于阴性菌,且表现为多重耐药;50个Ⅰ类整合子共检测到16种耐药基因盒,包括了编码氨基糖苷类耐药基因aadA1、aadA2、aac6-Ⅱ、aacA4,甲氧苄啶耐药基因drfA1、dfrA12、dfrA15、dfrA17、dfrB4,β内酰胺类耐药基因bla_(OXA-10)、bla_(OXA-21),氯霉素耐药基因catB3、catB8,利福平耐药基因arr2、arr3以及质粒介导的喹诺酮类耐药基因aac(6')-Ib-cr。携带了不同类耐药基因盒的整合子阳性菌还表现出所对应的对甲氧苄啶、链霉素、氯霉素类等的耐药表型,由此推测整合子的存在与细菌的多重耐药密切相关。研究表明,I类整合子分布于广东地区猪-鱼复合养殖模式下不同来源气单胞菌,并介导细菌对多种抗菌药物耐药。有必要开展畜禽-鱼复合养殖模式下细菌耐药性的传播机制研究,为水产养殖合理用药提供依据。     

斑点叉尾鮰养殖池塘底泥微生物群落结构特征及其影响因素

微生物群落在水产养殖环境中起着重要作用,深入了解微生物群落组成及其构建机制对了解池塘生态功能具有重要意义。本研究通过高通量测序技术研究斑点叉尾鮰（）养殖池塘底泥中细菌群落机构特征,并分析了细菌群落的主要影响因素。结果显示,斑点叉尾鮰养殖池塘底泥细菌群落结构呈现出季节性变化,冬春季样品相似性较高。斑点叉尾鮰养殖池塘底泥优势菌群主要为变形菌门（Proteobacteria）、绿弯菌门（Chloroflexi）、拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）和酸杆菌门（Acidobacteria）,与池塘水体细菌群落间有明显的差别。线性判别分析显示,冬、春和秋季的特异性优势菌群较为丰富,冬季和春季分布较为集中,分别集中于浮霉菌、拟杆菌门、绿弯菌门和厚壁菌门、拟杆菌门,秋季特异性优势菌群分布特别分散,而夏季优势细菌类群最少,只有绿弯菌门厌氧绳菌属1个属具有显著优势。环境因子中,透明度和总溶解性悬浮物与斑点叉尾鮰养殖池塘底泥细菌群落结构有显著相关（<0.05）,对细菌群落结构的影响也最大。本研究旨为斑点叉尾鮰池塘微生物群落的变化和调控提供借鉴。     

铜陵淡水豚国家级自然保护区浮游植物群落特征与水质评价

经济发展改变了铜陵淡水豚国家级自然保护区的水域水生生态环境,为了揭示该保护区浮游植物群落结构特征与水质状况,分别于2011年6月、7月及9月按对4个代表性断面进行浮游植物群落研究,并根据浮游植物群落结构的物种组成,及其结合Shannon-Weaver指数和 Pielou 均匀度指数对水质营养及污染状况进行评价。结果表明：该保护区浮游植物有86种,其中绿藻门最多,34种,占全部浮游植物种类的39.53%;硅藻次之,29种,占总种类的33.73%;蓝藻门14种,占总种类的16.28%。浮游植物总平均密度为9.79×105 ind•L-1,总平均生物量为0.303mg•L-1,调查区域浮游植物平均密度由高到低依次为铜陵沙头南>铜陵沙头北>萝卜洲尾南>萝卜洲尾北;平均生物量由高到低依次为铜陵沙头南>铜陵沙头北>萝卜洲尾北>萝卜洲尾南。铜陵淡水豚自然保护区水质属于富营养型,处于α-中污染状态。     

白菜亚硝酸还原酶基因BcNiR的克隆及表达分析

 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。     

Relationship between zinc and zinc transporter expression in human lung cancer cells

AIM: To study the changes of zinc transporter gene expression in A-549 cell line exposed to ZnCl2 and N,N,N’,N’-tetrakis 2-pyridylmethyl ethylenediamine (TPEN).METHODS: Human lung cancer cell line A-549 was exposed to different concentrations of ZnCl2 (0, 50, 100, 150, 200 μmol/L) and TPEN (0, 5, 10, 15, 20 μmol/L), respectively. Twelve hours later, the cell viability was measured by MTT (methyl thiazolyl tetrazolium) and levels of zinc transporter mRNA was detected by RT-PCR. Zinquin was used to estimate the intracellular zinc concentrations.RESULTS: A-549 cell viability rate was significantly decreased when exposed to ZnCl2 at concentrations of 150 and 200 μmol/L, and to TPEN. The intracellular zinc concentration was significantly increased when exposed to ZnCl2 and decreased when exposed to TPEN. Zinc transporter (ZnT-1) mRNA level was increased along with the increase in the concentration of ZnCl2 but decreased when exposed to TPEN. The expressions of ZIP-1 and ZIP-10 (Zrt-and Irt-like protein) were increased along with the increase in the concentration of TPEN but decreased when exposed to ZnCl2.CONCLUSION: ZnT-1 expression is induced by zinc supplement. ZIP-1 and ZIP-10 expressions are induced by zinc deficiency and repressed by zinc supplement.     

Effects of carvedilol on murine viral myocarditis

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.     

Impact of serial activation of embryonic pax2 and WT1 genes on transdifferentiation of renal tubular epithelial cells

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.     

Changes of immune function and mortality in patients with sepsis-induced stress hyperglycemia

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.     

Effects of salvianolic acid B on neurons during oxygen/glucose deprivation and reintroduction

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.