MiR-214 Targets β-Catenin Pathway to Suppress Invasion, Stem-Like Traits and Recurrence of Human Hepatocellular Carcinoma

The down-regulation of miR-214 has previously been observed in human hepatocellular carcinoma (HCC). Here, we demonstrated the down-regulation of miR-214 is associated with cell invasion, stem-like traits and early recurrence of HCC. Firstly, we validated the suppression of miR-214 in human HCC by real-time quantitative RT-PCR (qRT-PCR) in 20 paired tumor and non-tumor liver tissues of HCC patients and 10 histologically normal liver tissues from colorectal cancer patients with liver metastases. Further qRT-PCR analysis of 50 HCC tissues from an independent cohort of HCC patients of whom 29 with early recurrent disease (<2 years) and 21 with late recurrent disease demonstrated that the suppression of miR-214 was significantly more suppressed in samples from HCC patients with early recurrent disease compared those from patients with no recurrence. Re-expression of miR-214 significantly suppressed the growth of HCC cells in vitro and reduced their tumorigenicity in vivo. The enhancer of zeste homologue 2 (EZH2) and β-catenin (CTNNB1) was identified as two potential direct downstream targets of miR-214 through bioinformatics analysis and experimentally validated the miRNA-target interactions with a dual-firefly luciferase reporter assay. In corroborate with this, both EZH2 and CTNNB1 are found to be significantly overexpressed in human HCC biopsies. Since EZH2 can regulate CTNNB1, CTNNB1 can also be an indirect target of miR-214 through EZH2. Silencing EZH2 or CTNNB1 expression suppressed the growth and invasion of HCC cells and induced E-cadherin (CDH1), known to inhibit cell invasion and metastasis. Furthermore, the silencing of miR-214 or overexpression of EZH2 increased EpCAM(+) stem-like cells through the activation of CTNNB1. Interestingly, the up-regulation of EZH2, CTNNB1 and the down-regulation of CDH1 in HCC patients correlated with early recurrent disease and can be an independent predictor of poor survival. Therefore, miR-214 can directly or indirectly target CTNNB1 to modulate the β-catenin signaling pathway in HCC.     

Luteolin enhances TNF-related apoptosis-inducing ligand's anticancer activity in a lung cancer xenograft mouse model

Sensitization of cancer cells to apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by luteolin has been suggested by in vitro studies. However, no in vivo experiment has been reported to validate the potentiation effect of luteolin on TRAIL's anticancer activity. In this report, we first confirmed that luteolin potentiates TRAIL-induced cytotoxicity in A549 cells and HeLa cells in association with increased activation of apoptosis. Then we performed an in vivo experiment with a non-small cell lung cancer xenograft mouse model, which showed for the first time that the in vivo anticancer activity of TRAIL was greatly enhanced by luteolin. Compared with that in untreated control or treatment with TRAIL or luteolin alone, inhibition of tumor growth and apoptotic cell death in xenograft tumors were significantly increased in animals receiving combination treatment with TRAIL and luteolin. Data from this study thus provide strong in vivo evidence supporting that luteolin is a potential sensitizer for TRAIL in anticancer therapy.     

中国河南两株庚型肝炎病毒（HGV）NS5区部分cDNA的克隆与序列分析

通过逆转录－聚合酶链反应从我国河南省２例重叠感染ＨＣＶ或ＨＢＶ／ＨＤＶ的献血啼中,分离到ＨＢＶＮＳ５区的部分ｃＤＮＡ,对其进行序列分析比较,结果表明,河南株ＨＧＶＮＳ５工核苷酸与两中国ＨＧＶ主同源性高于国外代表株（８８．５－９０．６％）,但由核苷酸推导的氨基酸的同源性都无明显的地区性区别。ＨＧＶＮＳ５区氨基酸序列较保守,缺乏明显高变区,中国４株ＨＧＶ在７３８４位发生了由Ｃ→Ｔ的变异,从而导致一个人     

昆虫体内储存蛋白的研究进展

储存蛋白是昆虫体内普遍存在的一种特异性血淋巴蛋白 ,通常在幼虫的脂肪体内合成 ,释放进入血淋巴中。化蛹时 ,又被脂肪体选择性吸收 ,作为氨基酸的贮存库对成虫变态发育和雌性卵发育起着重要的作用。该文介绍了昆虫体内储存蛋白的特性、功能、及调节机制。     

根癌农杆菌介导Bt基因转化水稻的研究

为了培育出无筛选标记基因的转基因水稻,试验将loxp-hpt-loxp基因与成基因连锁在-起转化水稻方法,得到loxp-hpt—loxp—Bt转基因水稻植株,再与同质的带有ere基因的水稻杂交,以定向删除潮霉素抗性筛选标记。试验表明以水稻品种“皖粳97”为供试材料,将成熟胚来源的愈伤组织用根癌农杆菌EHA105／pCAMBIA1305．1感染后,筛选出抗性愈伤组织并获得再生植株。经PCR验证,得到20棵转基因水稻植株。     

日本对虾c型溶菌酶的高效重组表达及产物分析

从日本对虾(Marsupenaeus japonicus)血液中提取总RNA,根据GenBank已登录的该cDNA序列(AB080238),通过RT-PCR技术扩增出日本对虾溶菌酶(MjLys)成熟肽基因。该基因完整的开放阅读框为477 bp,编码158个氨基酸(aa),前18 aa为信号肽,成熟肽由140 aa组成,分子量为16.4 kD,理论等电点(pI)为8.80。经分析表明,该基因含有一个完整的c型溶菌酶结构域(1-130 aa),包括c型溶菌酶特有的两个活性中心Glu33和Asp50,以及8个保守结构Cys残基。将MjLys成熟肽基因亚克隆至原核表达载体pET-32a(+),在大肠杆菌细胞BL21(DE3)pLysS中诱导发酵,实现了重组MjLys蛋白的高效表达,并测定了该重组蛋白对几种细菌的抑菌活性。结果表明,重组日本对虾溶菌酶对革兰氏阳性菌金黄色葡萄球菌和溶壁微球菌均有显著的溶菌活性。     

Different Effects of Homocysteine and Oxidized Low Density Lipoprotein on Methylation Status in the Promoter Region of the Estrogen Receptor α Gene

We investigated the effects of homocysteine (Hcy) and oxidized low density lipoprotein (ox-LDL) on DNA methylation in the promoter region of the estrogen receptor α (ERos) gene,and its potentialmechanism in the pathogenesis of atherosclerosis.Cultured smooth muscle cells (SMCs) of humans weretreated by Hcy and ox-LDL with different concentrations for different periods of time.The DNA methylationstatus was assayed by nested methylation-specific polymerase chain reaction,the lipids that accumulated inthe SMCs and foam cell formations were examined with Oil red O staining.The proliferation of SMCs wasassayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.The results showedthat ox-LDL in moderate concentrations (10-40 mg/L) induced de novo methylation in the promoter regionof the ERα gene of SMCs.However,high concentrations (50 mg/L) of ox-LDL,resulted in demethylation ofERα.The Hcy treatment resulted in de novo methylation in the promoter region of the ERα gene with aconcentration- and treating time-dependent manner,and a dose-dependent promoting effect on SMCproliferation.These data indicated that the two risk factors for atherosclerosis had the function of inducingde novo methylation in the promoter region of the ERα gene of SMCs. However,high concentrations (50rag/L) of ox-LDL induced demethylation,indicating that different risk factors of atherosclerosis with differentpotency might cause different aberrant methylation patterns in the promoter region of the ERα gene.Theatherogenic mechanism of Hcy might involve the hypermethylation of the ERα gene,leading to the proliferationof SMCs in atherosclerotic lesions.     

水氮运筹对膜下滴灌棉花光合特性及产量形成的影响

以不同基因型棉花品种为材料,在土柱栽培条件下研究膜下滴灌条件下水氮运筹方式对新疆棉花光合性能和产量构成的影响.结果表明： 播前灌溉+盛花期前限量滴灌+盛花期后充分滴灌,并配合氮肥基施20%+追施80%的水氮运筹方式（W₄N₂）下,盛花期叶片叶绿素含量、气孔导度（g_s）、净光合速率（P_n）、PSⅡ实际光化学效率（Φ_PSⅡ）和光化学猝灭系数（q_P）均显著低于全生育期常规滴灌处理,非光化学猝灭系数（NPQ）增加,地上部干物质累积量受到限制;盛铃期至吐絮期叶绿素含量、g_s、P_n、Φ_PSⅡ、q_P均随水氮供应量的提高而增大,地上部干物质产生超补偿积累,且有利于光合产物向棉铃的运转与分配.在氮肥基施20%+追施80%的施氮方式下,新陆早13号以播前灌溉+全生育期常规滴灌（W₃）处理的籽棉产量较高,新陆早43号以播前灌溉+盛花期前限量滴灌+盛花期后充分滴灌（W₄）处理籽棉产量最高.因此,在播前灌溉条件下适当减少盛花期前、增加生育中后期水氮供应,可以延长冠层叶片光合功能期,促进光合物质优先向生殖器官分配,充分发挥膜下滴灌棉花的增产潜力.     

热带季节雨林林窗边缘不同热力作用面热力效应的季节变化特征

通过对热带季节雨林雾凉季和湿热季昼间林窗区域不同热力作用面的热力效应初步分析,指出在西双版纳,不论是雾凉季还是湿热季,热带季节雨林林窗边缘壁面均具有不可忽视的热力作用,且由于受林缘树木的影响,热力效应较强的东侧,北侧林缘壁面最大区域出现位置高于次生林林窗,而强度小于次生林林窗,显示了林窗边缘壁面的热力效应除与太阳高度角,太阳辐射的时间长短和强度有关之外,林窗边缘树木高度也是不可忽视的因子,其结果可为进一步研究林窗小气候形成机制提供研究基础,为研究林窗更新及生物多样性问题提供科学参考。     

哌替啶对心室肌收缩的抑制作用及其机制

为明确哌替啶对心脏收缩的直接效应 ,并探讨其相关机制。采用Langendorff灌流心脏模型 ,观察了哌替啶对大鼠心室收缩功能的影响 ,并用荧光测钙技术和膜片钳技术探讨了哌替啶作用的钙离子机制。结果显示 ,哌替啶剂量依赖性地降低离体灌流心脏的LVDP×HR、 +dP/dt和 -dP/dt,而升高LVEDP。在酶解分离的心室肌细胞上 ,哌替啶剂量依赖性地降低细胞收缩时的钙瞬变幅度 ,并升高舒张末期的钙水平。哌替啶不影响高浓度咖啡因诱导的内钙释放。哌替啶使L 型钙电流强度降低到给药前的 67 4± 10 1% ,而不改变钙通道的激活和失活电位。哌替啶减弱钙电流的作用并不能被阿片受体阻断剂纳洛酮所阻断。以上结果表明 ,哌替啶能通过非阿片受体介导的途径阻断细胞外钙离子的内流 ,对心室收缩产生直接的抑制作用