shRNA‑mediated knockdown of KNTC1 inhibits non-small-cell lung cancer through regulating PSMB8

In view of the important roles played by Kinetochore proteins in mitosis, we believed that they may contribute to the development and progression of human cancers, which has been reported recently elsewhere. Kinetochore-associated 1 (KNTC1) participates in the segregation of sister chromatids during mitosis, the effects of which on non-small-cell lung cancer (NSCLC) remain unclear. Here, we sought to identify the biological significance of KNTC1 in NSCLC. KNTC1 protein expression in NSCLC tissues was investigated by immunohistochemistry. Lentivirus delivered short hairpin RNA (shRNA) was utilized to establish KNTC1 silence NSCLC cell lines. The effects of KNTC1 depletion on NSCLC cell proliferation, migration, apoptosis, and tumor formation were analyzed by MTT assay, wound-healing assay, transwell assay, flow cytometry assay, and in nude mouse models in vivo. After KNTC1 reduction, NSCLC cell viability, proliferation, migration, and invasion were restrained. A xenograft tumor model was also provided to demonstrate the inhibited tumorigenesis in NSCLC. In addition, the downstream mechanism analysis indicated that KNTC1 depletion was positively associated with PSMB8. The findings of the present study suggested that KNTC1 may have a pivotal role in mediating NSCLC progression and may act as a novel therapeutic target for NSCLC.Subject terms: Non-small-cell lung cancer, Cell migration     

转双抗虫基因107杨节肢动物群落特征及相对稳定性

于2015年5—9月对唐山市滦南县苗圃场内的转双抗虫基因(Bt Cry1Ac和API基因)107杨(简称:转抗虫基因107杨)及未转基因107杨(简称:对照杨)进行了节肢动物群落调查,利用群落特征指数、群落相似性系数、群落相对稳定性和主成分分析法对转抗虫基因107杨节肢动物群落特征、相似性、相对稳定性及主成分进行对比分析。调查共获得节肢动物6818头,隶属于2纲,8目,43科,58种。研究结果表明:转抗虫基因107杨和对照杨节肢动物群落中以鳞翅目、鞘翅目、膜翅目、双翅目昆虫为主要类群,其中鳞翅目昆虫个体数量最多;鳞翅目和半翅目昆虫个体数量与对照杨差异显著;在功能类群上,转抗虫基因107杨食叶昆虫个体数量较对照杨显著减少,刺吸昆虫个体数量显著增加;转抗虫基因107杨节肢动物群落多样性指数、均匀度指数较高,优势度指数较低,节肢动物群落物种分布较均匀;相似性结果显示,物种组成与对照杨相似度较高;转抗虫基因107杨节肢动物群落物种间在数量上的制约作用较强;主成分分析表明食叶昆虫物种数量与个体数量、刺吸昆虫物种数量和其他植食性昆虫个体数量是影响转抗虫基因107杨与对照杨节肢动物群落变化的共同主导因子。     

植被恢复过程中芒萁覆盖对侵蚀红壤氮组分的影响

氮素是限制陆地生态系统生产力的重要因子。采用时空代换法,以红壤侵蚀区未治理、恢复12年和30年的马尾松林为研究对象,对比分析了林下芒萁覆盖地与裸地表层土壤之间氮同位素、不同形态氮组分含量以及不同组分氮含量所占比例之间的差异。结果表明:在所有马尾松林中,芒萁覆盖增加了表层土壤的全氮含量,δ~(15)N值则比林下裸地显著降低了33. 8%—83.1%(P0.05)。随着恢复年限增加,林下芒萁覆盖地表层土壤δ~(15)N值显著下降,而林下裸露地δ~(15)N值没有显著变化(P0.05)。不同恢复年限马尾松林的芒萁覆盖地表层土壤微生物生物量氮、可溶性有机氮和铵态氮含量显著高于林下裸地(P 0.05),而硝态氮含量则显著低于林下裸地(P0.05)。随恢复年限增加,表层土壤微生物生物量氮、可溶性有机氮、铵态氮含量均呈增加趋势,而硝态氮含量则呈下降趋势,不同形态氮占全氮比例表现为:微生物生物量氮铵态氮可溶性有机氮硝态氮。相关分析表明土壤δ~(15)N值与硝态氮极显著正相关,与其他氮组分极显著负相关(P0.01)。由此可见,与林下裸地相比,芒萁覆盖在植被恢复过程中有助于提高表层土壤中全氮、微生物生物量氮、可溶性有机氮和铵态氮含量,降低硝态氮的淋溶损失风险,促进土壤氮保持和积累,从而有利于退化红壤生态系统的恢复。     

MYB 转录因子在植物抗逆胁迫中的作用及其分子机理

摘要: 在植物抗逆反应体系中, 通过转录因子调控功能基因的表达, 是植物逆境应答反应的关键环节。作为植物体内最大的转录因子家族之一, MYB(v-myb avian myeloblastosis viral oncogene homolog)类转录因子在植物抗逆胁迫过程中起着重要的作用。文章论述了MYB转录因子的结构特征、功能特性及其分子机理, 并结合研究进展着重讨论了它们在植物抗逆胁迫中的作用。     

上海部分地区戊型肝炎病毒(HEV)基因型的分析

为了解戊型肝炎病毒(HEV)在上海部分地区流行的基因型,采用RT-nPCR的方法检验35例急性散发性戊型肝炎患者中HEV RNA,并对阳性产物进行克隆测序,然后对其基因型进行分析.结果显示在35例急性散发性戊型肝炎患者中PCR阳性为9例,测序证实8例为HEV的基因序列;其中1例为HEV 1型,7例为HEV 4型.提示在上海部分地区的急性散发性戊型肝炎中以HEV 4型感染为主.     

Characterization and expression of the plasmid-borne bedD gene from Pseudomonas putida ML2, which codes for a NAD+-dependent cis-benzene dihydrodiol dehydrogenase.

The catabolic plasmid pHMT112 in Pseudomonas putida ML2 contains the bed gene cluster encoding benzene dioxygenase (bedC1C2BA) and a NAD+-dependent dehydrogenase (bedD) required to convert benzene into catechol. Analysis of the nucleotide sequence upstream of the benzene dioxygenase gene cluster (bedC1C2BA) revealed a 1,098-bp open reading frame (bedD) flanked by two 42-bp direct repeats, each containing a 14-bp sequence identical to the inverted repeat of IS26. In vitro translation analysis showed bedD to code for a polypeptide of ca. 39 kDa. Both the nucleotide and the deduced amino acid sequences show significant identity to sequences of glycerol dehydrogenases from Escherichia coli, Citrobacter freundii, and Bacillus stearothermophilus. A bedD mutant of P. putida ML2 in which the gene was disrupted by a kanamycin resistance cassette was unable to utilize benzene for growth. The bedD gene product was found to complement the todD mutation in P. putida 39/D, the latter defective in the analogous cis-toluene dihydrodiol dehydrogenase. The dehydrogenase encoded by bedD) was overexpressed in Escherichia coli and purified. It was found to utilize NAD+ as an electron acceptor and exhibited higher substrate specificity for cis-benzene dihydrodiol and 1,2-propanediol compared with glycerol. Such a medium-chain dehydrogenase is the first to be reported for a Pseudomonas species, and its association with an aromatic ring-hydroxylating dioxygenase is unique among bacterial species capable of metabolizing aromatic hydrocarbons.     

人谷胱甘肽硫转移酶A1在乳酸乳球菌中的表达及活性研究

用RTPCR技术从人肝总RNA中分离扩增了人谷胱甘肽硫转移酶A1基因的cDNA序列,克隆至大肠杆菌表达质粒pET23b,采用蛋白表达筛查法及DNA测序证明该cDNA序列完全正确。重组质粒pET23bhgst转化大肠杆菌BL21(DE3),经IPTG诱导获得高效表达的可溶性hGSTA1产物,其表达量约为大肠杆菌可溶性总蛋白的40%。将hGSTA1cDNA亚克隆至乳酸乳球菌表达载体pMG36e,电穿孔法转化乳酸乳球菌MG1363获得hGSTA1乳酸乳球菌表达株。SDSPAGE及Western杂交分析表明该菌株表达预期大小的hGSTA1融合蛋白,经谷胱甘肽琼脂糖亲和层析纯化获得的hGSTA1蛋白具有较高的谷胱甘肽硫转移酶活性。具hGSTA1酶活性的乳酸乳球菌工程菌可望应用于研制防癌保健乳制品。     

鼎湖山森林生态系统演替过程中的能量生态特征

以时空替代的方法,将灌草丛、针叶林、针阔叶混交林和季风常绿阔叶林等４个处于同一空间下的群落当作同一样落演替进程中的４个阶段,研究了鼎湖山南亚热带森林演替过程中的能量生态特征。结果表明,鼎湖山南亚热带森林群落演替过程中,其垂直层次、叶面积指数、冠层对太阳辐射能的截获量、叶生物量、总生物量、总初级生产力、总呼吸量、净初级生产力、枯树木现存量和年输入量、昆虫啃食量、群落的能量现存量等随演替的进程而增加,     

PFGE在真菌基因研究中的应用

IPFGE与连锁群从酿酒酵母脉冲电场电泳研究到医学上重要酵母类真菌的分类,人们已逐渐建立了参数随菌种各异的PFGE电泳条件,以适于不同大小染色体的分离,获得了一系列酵母类及丝状真菌,包括四大纲以及非典型真菌的染色体核型及基因组大小（表1）,并利用已知基因探针进行染色体连锁群的定位。1987年,Sndth等l’吩离得到了粟酒裂殖酵母的3条巨大染色体,使PFGE的分辨能力达到Tgmb,极大促进了该菌株的基因图谱工作的进展。紧接着,他们又得到了假丝酵母类具有种特异性的电泳核型,并成为分子流行病学中酵母类致病真菌的分型依据。…