Effect of idazoxan on permeability of inflammatory blood-brain barrier model in vitro

AIM: To study the effect of idazoxan on the permeability of inflammatory blood-brain barrier (BBB) model in vitro and the expression of tight junction protein ZO-1.METHODS: In vitro BBB model was established by murine brain endothelial cell line bEnd.3 incubated for 7 d. The cells were treated with TNF-α (10 nmol/L) for additional 24 h to establish the inflammatory BBB model, which was pretreated with IDA at doses of 50, 100 and 200 μmol/L, respectively. The permeability was measured using fluorescein isothiocyanate-conjugated dextran (FD-40, MW 40,000), the expression of ZO-1 was detected by Western blot analysis, the distribution of ZO-1 was observed by immunofluorescence, and the mRNA expression of MMP-9/TIMP-1 was measured by RT-PCR.RESULTS: After incubated for 7 d, b.End3 cells converged to be confluent monolayer with low permeability. The inflammatory BBB model induced by TNF-α treatment displayed much higher permeability with decreased expression of tight junction protein ZO-1, destroyed distribution of ZO-1 and increased mRNA expression of MMP-9. When pretreated with IDA, the permeability was greatly decreased, the expression of ZO-1 was greatly increased, the abnormal distribution of ZO-1 was greatly ameliorated and the mRNA expression of MMP-9 was obviously reduced. The effect was most significant in IDA (200 μmol/L)-pretreated group (P<0.01).CONCLUSION: IDA directly acts on brain endothelial cells to reduce the expression of MMP-9, increase the expression of tight junction protein ZO-1 and ameliorate the destroyed distribution of ZO-1 in the inflammatory BBB, thus reversing the abnormally elevated permeability in a inflammatory BBB model in vitro induced by TNF-α.     

Expression of telomerase inhibitor Pinx1 in leukemia cells and its correlation with telomerase activity

AIM:To study the expression of telomerase inhibitor Pinx1 in acute leukemia cells and during the differentiation of acute promyelocytic leukemia cells,and to realize its effect on telomerase activity.METHODS:Realtime quantitative PCR with fluorescence probe hybridization was used to measure the expression of Pinx1 and hTERT mRNA in acute leukemia cells and during differentiation of NB4 cells induced by ATRA.The correlations between Pinx1 and hTERT expression were also analyzed.RESULTS:Pinx1 mRNA expression in acute leukemia samples (0.00312,5.42×10-4-0.024) was significantly higher than that in normal bone marrow mononuclear cells (7.89×10-4,0-0.00863,P<0.01).The expression of Pinx1 mRNA had significant positive correlation with hTERT mRNA expression (r=0.296,P<0.05).Pinx1 mRNA expression decreased during differentiation,its expression was positive correlated with hTERT mRNA expression (r=0.900,P<0.05).CONCLUSIONS:As an inhibitor of telomerase,however,Pinx1 also had the same direction of regulation with telomerase activity in acute leukemia cells,suggesting its expression variation may be a subsequent reaction induced by that of hTERT to stabilize telomerase activity.The exact mechanisms remained to be verified.     

光明顶上种核桃 远近闻名小康村——云南省大理州漾濞彝族自治县苍山西镇光明村

"头顶云雾脚踩霜,荞麦粑粑萝卜汤"这曾是旧光明村人生活的真实写照。光明村位于漾濞彝族自治县东部,苍山西坡腹地,居住着彝、汉、白、傣、傈僳5个民族,少数民族人口占总人口的65.04%。       

剪除部分颖壳对宁夏水稻糙米生长发育的影响

在温室盆栽条件下,以粳稻普通稻宁粳28号和优质稻宁粳38号为材料,于开花后两天设剪壳、剪壳后套硫酸纸袋保湿透光、剪壳后套硫酸纸袋外再套牛皮纸袋保湿挡光及不剪壳4个处理,研究谷壳对糙米生长发育的影响.结果表明:①剪壳处理的糙米千粒质量低于不剪壳处理,但差异不显著.随着糙米生长发育进程的推进,差异显著性增大;②剪壳后套硫酸纸袋处理糙米的千粒质量显著高于不剪壳处理,并随着生长发育的推进,其差异增大;③剪壳后套硫酸纸袋再套牛皮纸袋处理的糙米千粒质量显著高于剪壳后套硫酸纸袋;④剪壳、剪壳后套硫酸纸袋、外再套牛皮纸袋处理对普通稻宁粳28号糙米千粒质量的影响大于优质稻宁粳38号;⑤谷壳对粳稻普通稻宁粳28号糙米生长发育的影响比对优质稻宁粳38号的明显.     

利用CRISPR/Cas9敲除葡萄VviPDS1基因的研究

选择葡萄八氢番茄红素脱氢酶(Phytoene desaturase)基因VviPDS1为靶标,利用CRISPR/Cas9系统构建基因敲除载体,瞬时转化葡萄叶片原生质体,检测到不同类型的突变。通过农杆菌介导转化‘无核白’葡萄胚性愈伤组织,筛选获得卡那霉素抗性植株71株。经PCR鉴定,其中53株为阳性植株,阳性率为74.64%。测序结果表明,共有20株在靶点发生不同类型的突变,编辑效率为37.74%;其中9株产生了双等位基因突变。对其进行氨基酸序列预测,在第202位氨基酸之后发生了不同程度的变异。利用CRISPR/Cas9系统敲除VviPDS1获得的突变体植株呈现整体矮化,其叶片出现不同程度白化。表明CRISPR/Cas9系统可以通过细胞中的瞬时或稳定表达进行基因编辑,可以实现在葡萄编辑植株中产生纯合敲除。     

Serum contents of E-selectin,sICAM-1, sVCAM-1 in patients with acute coronary syndrome

AIM:To explore the serum levels of certain adhesion molecules and its significance in acute coronary syndrome(ACS). METHODS:The subjects included 40 patients with acute myocardial infarction(AMI) and 40 patients with unstable angina pectoris (UAP). Among the 80 patients, 60 patients accepted a follow-up 4 months. At the same time we selected 40 controls from people who attended a routine health check in the university. Serum levels of E-selectin, sICAM-1, sVCAM-1 were measured by ELISA.RESULTS:Serum levels of E-selectin, sICAM-1, sVCAM-1 were significantly higher in the ACS group(AMI or UAP) than in the control group. Four months later, the levels of E-selectin, sICAM-1 became significantly lower in the follow-up group than in the ACS group, while sVCAM-1 showed no significant difference. CONCLUSION:Serum levels of E-selectin, sICAM-1 may have certain diagnostic value for ACS, and can be a useful marker reflecting the stability of the disease.     

Ss-Bi1 encodes a putative BAX inhibitor-1 protein that is required for full virulence of Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a destructive necrotrophic plant pathogen with global distribution. Although S. sclerotiorum has been studied extensively, substantial research on aspects of the pathogen's ability to cause disease is still needed. Bax inhibitor-1 protein functions as a suppressor of programmed cell death and is involved in the response to biotic and abiotic stress in animals, plants and yeast. In this study, we functionally characterized a putative Bax inhibitor-1 protein, Ss-Bi1, from S. sclerotiorum. Ss-Bi1 is predicted to contain a BAX inhibitor-1-like super family domain and shows significant homology with many BAX inhibitor-1 proteins. High expression levels of Ss-Bi1 were observed in hyphae under various stresses. Targeted silencing of Ss-Bi1 resulted in reduced virulence in host plants. Ss-Bi1 gene-silenced strains were more sensitive to heat stress and ER stress than the wild-type strain. The results suggest that Ss-Bi1 encodes a putative BAX inhibitor-1 protein that is required for full virulence of S. sclerotiorum.     

Combination of dihydromyricetin and cisplatin significantly induces overexpression of Noxa and Bim in PC3 cells

AIM:To investigate the adjuvant effect of dihydromyricetin on cisplatin-based chemotherapy in prostate cancer and its mechanisms. METHODS:The viability of LNCaP and PC3 cells treated with different concentrations of dihydromyricetin and cisplatin was measured by MTT assay. The expression of FOXO1, Noxa and Bim, release of cytochrome C from mitochondria, and activation of caspase-9 and caspase-3 in PC3 cells treated with dihydromyricetin and cisplatin were determined by Western blot. Co-immunoprecipitation was performed to detect the interaction of apoptotic protease-activating factor-1 (Apaf-1) and caspase-9 in the PC3 cells. The apoptotic rate of PC3 cells was analyzed by flow cytometry. RESULTS:Adjuvant therapy of dihydromyricetin significantly enhanced the anti-tumor effect of cisplatin against prostate cancer in vitro. Dihydromyricetin significantly promoted the expression of FOXO1 in the PC3 cells. However, transfection with FOXO1 small interfering RNA (siRNA) obviously suppressed the adjuvant effect of dihydromyricetin. Combination of cisplatin and dihydromyricetin significantly induced the overexpression of Noxa and Bim, the release of cytochrome C, the interaction of Apaf-1 and caspase-9, the activation of caspase-9 and caspase-3, and the apoptosis in the PC3 cells. On the other hand, transfection with FOXO1 siRNA obviously suppressed the apoptotic pathway of PC3 cells treated with dihydromyricetin and cisplatin. CONCLUSION:Dihydromyricetin enhances the cytotoxicity of cisplatin against prostate cancer through the FOXO1-Bim/Noxa pathway in vitro.     

景东烟区生态因素与烤烟质量特点分析

为了探讨景东烟区生态因素与烤烟质量之间的关系,主要对云南景东烟区的植烟气候因素、土壤养分状况以及所产烤烟的质量特点进行了分析。结果表明:景东烟区具有无霜期长,大田期气温适宜,光照充足,降雨充沛,雨热同步等特点;植烟土壤pH值适宜,有机质含量较丰富,具有较强的保肥能力,土壤碱解氮含量高,速效磷、钾含量适中;交换镁含量较高,速效锌、硼含量缺乏。烤烟外观质量较好,香气量较足、香气质较好,燃烧性强,但在化学成分间的协调性方面不及津巴布韦烤烟。     

不同生长阶段尼罗罗非鱼IGF1基因表达分析

为阐明IGF1基因在罗非鱼生长过程中的表达规律,为罗非鱼的分子选育等工作奠定了理论基础,本实验利用实时荧光定量PCR技术跟踪尼罗罗非鱼(Oreochromis niloticus)在不同生长阶段（0、20、40、60、80、125、145、165、185、205天）肝脏和肌肉胰岛素样生长因子1(IGF1)基因表达的发育性变化。结果表明：实验鱼的体重增长过程可以分为快速生长期和平稳期。在快速生长期时,肝脏IGF1基因表达水平较高,平稳期时较低,肝脏中IGF1基因表达情况与体重增长趋势基本吻合。肌肉中IGF1基因mRNA含量要低得多,而且表达模式与肝脏IGF1基因不相同。研究结果提示：IGF1 mRNA的表达具有明显的时空变化和组织特异性。肝脏中IGF1基因的表达量变化与罗非鱼体重增长趋势一致。肌肉中IGF1表达模式与肝脏中不同,提示肌肉中IGF1的分泌和作用有其自身的特异性。