14-3-3σ negatively regulates Akt and inhibits Rat1-Akt cell xenografts

AIM:The present study was designed to investigate the effect of adenovirus-mediated 14-3-3σ on Rat1-Akt cell xenografts and to explore whether the effect was mediated through negative regulation of Akt. METHODS:The effect of Ad-14-3-3σ on Rat1-Akt cells xenografts was observed in nude mice ex vivo and in vivo. Western blotting was used to detect 14-3-3σ protein,phospho-Akt (Thr 308), phospho-Akt (Ser 473), and phospho-(Ser/Thr) Akt substrate in tumor tissue after transfection of 14-3-3σ gene. RESULTS:The tumor volume was dramatically decreased and its emergence was delayed, regardless of using Ad-14-3-3σ-treated Rat1-Akt cells (ex vivo) or injected Ad-14-3-3σ in vivo, of which the effect of the continuously injected group was the best. Levels of Akt protein, phosphorylated Akt and phosphorylated Akt substrates in tumors obtained from Ad-14-3-3σ-treated mice were markedly less than those in PBS or Ad-β-gal-treated mice. CONCLUSION:14-3-3σ suppressed Akt overexpession in cells of Rat1-Akt xenografts by negatively regulating Akt.     

Detection and analyses of coagulation and fibrolysis factor mRNA levels in tissue of hepatocellular carcinoma patients

AIM:To detect the mRNA expression of tissue factor (TF), urokinase-type plasminogen activator (u-PA) and urokinase-type plasminogen activator receptor (u-PAR) in the samples of hepatocellular carcinoma (HCC) and adjacent tissue, and to elucidate their association with clinical significance. METHODS:The mRNA levels of TF, u-PA and u-PAR in 27 human HCC tissue samples with their adjacent tissue samples and 27 normal liver tissues were detected by RT-PCR. The relationship between the mRNA expression and their clinic-pathological data were also analyzed. RESULTS:The expressive rate of TF, uPA and uPAR in HCC tissue samples was 62.96% (17/27), 70.37% (19/27), 77.78% (21/27), respectively and relative expression intensity of TF, uPA and uPAR were 0.567±0.268, 0.964±0.458 and 0.784±0.322, respectively, which were significantly higher than those in adjacent tissue group and normal liver group (P<0.05). TF, uPA and uPAR mRNA levels were all significantly higher in larger size and in the invasive and metastatic subgroup (P<0.05). There had positive relationship between the mRNA expressions of these three factors. CONCLUSION:The results indicate that TF, uPA and uPAR might play an important role in hepatic carcinogenesis, invasiveness and metastasis.     

Gelatinases expression and their activity in rat alveolar macrophages induced by cigarette smoke medium

AIM: To study the effect of cigarette smoke medium (CSM) on the gene expression and activity of gelatinases from alveolar macrophages (AMs) in the rat, and then to explore their role in the pathogenesis of chronic obstructive pulmonary disease (COPD). METHODS: AMs were obtained from BALF of the rats that had smoked for 12 weeks. CSM was produced following the method of Wirtz and colleagues, and the cultured AMs were respectively stimulated for 24 h by 0%, 1%, 3%, 5%, 10%, 15% CSM. The mRNA levels of MMP-9 and MMP-2 were detected by semi-quantitative RT-PCR, and the enzyme activity was measured by Zymography. RESULTS: When the concentration of CSM was below 5%, the expression and activity of MMP-9 and MMP-2 signficantly increased with the concentration of CSM in a dose-depended manner (P<0.05). While the concentration of CSM exceeded 5%, the expression and activity of MMP-9 and MMP-2 correspondingly decreased with the increase in CSM concentrations (P<0.05), which possibly were related with the cytotoxicity of CSM. CONCLUSION: The expression and activity of MMP-9 and MMP-2 are induced by CSM. The gelatinases induced by smoking from AM may play an important role in the pathogenesis of COPD.     

Hepatocyte growth factor protects hepatocytes from apoptosis induced by actinomycin D

AIM: To investigate the anti-apoptotic effect of hepatocyte growth factor (HGF) on actinomycin D (ActD)-induced apoptosis in hepatocytes and the possible pathway. METHODS: Hepatocytes were exposed to ActD and HGF. The cytotoxic effects of ActD were tested by MTT. Apoptotic cells were identified by Hoechst 33342 staining, flow cytometry and detection of DNA fragmentation with agarose gel. Akt and phospho-Akt were detected by Western blotting analysis. RESULTS: The results showed that ActD induced apoptosis in hepatocytes 5 h after treatment. Phosphatidylinositol-3 kinase (PI-3K) specific inhibitor wortmannin enhanced the apoptotic effect of ActD. Furthermore, HGF significantly reduced the apoptosis in hepatocytes induced by ActD in a concentration-dependent manner. In the presence of wortmannin, HGF did not overcome apoptosis. CONCLUSION: Wortmannin enhances the apoptotic effect of ActD. HGF protects hepatocytes from apoptosis induced by ActD through a PI3K/Akt pathway.     

Exogenous carbon monoxide protects against the lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To study the mechanism of protective effect of exogenous carbon monoxide (CO) in the lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats. METHODS: Thirty-two SD rats were randomly divided into 4 groups: control, control+CO, IR and IR+CO. A rat model of ischemia in hind limbs and the reperfusion lung injury was made. The rats in IR+CO and control+CO groups were exposed to air containing 2.5×10-8 CO for 1 h before reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The lung tissue structure, polymorphonuclear leukocyte (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content and the animal survival rate were observed. The carboxyhemoglobin (COHb) levels in artery blood were detected with CO-oximeter and the expression of intercellular adhesion molecule-1 (ICAM-1) in the lung was detected by Western blotting. RESULTS: Compared to control, the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly increased in IR group. Compared with the IR group, the blood COHb level was significantly increased and the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly decreased in IR+CO group. CONCLUSION: These data suggest that exogenous CO attenuate limb IR-induced lung injury by down-regulatiny ICAM-1 expression and suppressing PMN sequestration in the lung following limb IR in rats.     

中国锦紫苏类病毒的检测及其分子生物学特征研究

 从无明显症状表现的11株锦紫苏叶片中抽提低分子量的RNA,经Return-PAGE、RT-PCR和Dot-blot hybridization检测,结果表明11株锦紫苏全部带有锦紫苏类病毒(Coleus blumei viroid,CBVd)。将部分PCR产物克隆到pGEM-3Zf(+)载体上并进行DNA序列测定。序列分析结果,所克隆的序列(GenBank登录号分别为DQ178395、DQ178396、DQ178397、DQ178398和DQ178399)与GenBank中报道的锦紫苏类病毒1号(CBVd-1)序列同源性为85.23%~99.20%。从市场上购买的锦紫苏种子经Return-PAGE和RT-PCR检测不携带锦紫苏类病毒。这是中国发生的锦紫苏类病毒的首次报道。     

纳他霉素防治苹果贮运期病害的初步研究

采用生长速率法测定了纳他霉素对苹果轮纹病菌、苹果黑星病菌、苹果黑斑病菌、苹果腐烂病菌的室内毒力,结果表明,该制剂对各病菌的抑制有效中浓度EC50分别为18.44、49.43、32.08、29.87μg/mL,并用病原菌感染方法测定了制剂对贮运期苹果的实物防治效果。纳他霉素浓度为200μg/mL的制剂各处理组在10d、14d、18d的防效分别达到52.91%、55.02%、54.12%,显著高于多菌灵对照的防效。     

大豆锈病研究现状与进展

大豆锈病近年在东南亚、南美洲发展迅速,对大豆生产威胁越来越大,尤其2004年11月登陆美国大陆,更引起大豆研究者、大豆生产国和进口国的极大关注。本文介绍了近年国际上大豆锈病的研究进展,主要包括大豆锈病的发现、地理分布、病原、症状、寄主范围、病原菌生理分化和防治技术研究等。     

荔枝果皮褐变程度的数学模型研究(英文)

为了明确荔枝采后果皮褐变的程度,为生产上的分级和研究中定量果皮的褐变程度提供依据,以淮枝、糯米糍、桂味为材料,研究了采后其果皮颜色变化过程中亮度(L*)、色饱和度(C*)、色度角(h)值及a*值和b*值的变化规律,探讨这些指标与褐变程度的关系,并推导出褐变与这些指标变化相关的方程式。结果表明,色度指标L*,a*,b*和C*值的变化均可用于衡量荔枝果皮的褐变程度,其中以C*值的相关性最高,并确定了3个品种荔枝C*值与褐变程度的对应关系,将荔枝果皮褐变程度数量化,建立荔枝果皮褐变程度的数学模型。