Artemisinin-derived dimer phosphate esters as potent anti-cytomegalovirus (anti-CMV) and anti-cancer agents: A structure–activity study

We recently reported the anti-cancer and anti-cytomegalovirus (CMV) activity of artemisinin-derived trioxane diphenylphosphate dimer 838. To probe the relationship between chemical structure and anti-CMV and anti-cancer activities, we now report synthesis and evaluation of a series of eight new dimer phosphate ester analogs of 838. This series of novel molecules was screened against human foreskin fibroblasts (HFFs) infected with CMV and against the human Jurkat T cell acute lymphoblastic leukemia cell line. This SAR study confirms the very high anti-CMV and anti-cancer potencies of dimer diphenyl phosphate ester 838 without its being toxic to normal cells.     

黄土高原羊圈沟小流域人工物种和自然物种径向生长对气候变化的响应差异

为揭示黄土高原人工和自然物种径向生长对气候变化的响应差异,在延安羊圈沟小流域分别获取人工和自然物种的树木年轮材料并构建标准年表,其中人工物种为刺槐(Robinia pseudoacacia)和柠条(Caragana korshinskii),自然物种为山杏(Armeniaca sibirica)和荆条(Vitex negundo var.heterophylla),并对年表中的气候信号进行了统计分析。结果表明:1)人工物种年表中的气候信号较强,主要表现在5—8月份,与温度呈负相关关系(刺槐:r=-0.427—-0.511,P0.05;柠条:r=-0.227—-0.738,P0.05),与降雨则呈正相关关系,但相关系数未达到显著性水平;自然物种年表中的气候信号较弱,与温度和降雨的相关关系均较低;2)不同于自然物种,人工物种树轮年表还与去年夏季(7—9月份)温度(负相关)和降水(正相关)存在相关关系,表明人工物种树木生长对气候因子存在一定滞后性;3)人工物种树轮年表与PDSI干旱指数在各月份均维持正相关关系,在生长季(刺槐4—9月、柠条4—8月)达到显著水平(刺槐:r=0.481—0.704,P0.05;柠条:r=0.314—0.610,P0.05);而自然物种年表与PDSI干旱指数的相关关系较弱,均未达到相关性水平。从各年表与气候要素(温度、降雨、PDSI)响应强度来看,黄土高原人工物种树木生长受水分胁迫显著,且以刺槐最为明显,其次是柠条;自然物种树木生长则没有明显干旱胁迫的影响,仅山杏生长受一定水分胁迫影响,荆条生长则与各气候要素关系较弱,水分胁迫对其生长的影响已很小。本研究的结果表明黄土高原人工物种生长明显受到水分条件限制,而自然恢复物种生长则受水分条件影响较小,能适应黄土高原干旱半干旱气候条件。     

Enhancement of adenosine production by Bacillus subtilis CGMCC 4484 through metabolic flux analysis and simplified feeding strategies

The objective of this research was to understand how the initial glucose concentration influences adenosine (AR) production and metabolic flux shift on the cultivation of Bacillus subtilis CGMCC 4484. Experiments confirmed that initial glucose concentration affects cell growth, AR production and metabolites, significantly. The flux distribution at the key nodes of glucose-6-phosphate (G6P), pyruvate (PYR) and acetyl coenzyme-A (AcCoA) could be affected by changing the glucose concentration. Based on kinetic analysis of specific rates, the low-glucose concentration was better for both cell growth and AR production during the first 12 h. However, the high-glucose concentration was more favorable for AR formation after 18 h. Furthermore, different simplified feeding strategies were designed to achieve higher AR accumulation. The final AR concentration of 15.60 g L^?1 was achieved when an optimized constant-feeding strategy was used, which was 21.02 % higher than batch fermentation. This was the first time to investigate the regulation of the glucose metabolism of AR-producing B. subtilis.     

Myocardin and microRNA‐1 modulate bladder activity through connexin 43 expression during post‐natal development

Overactive bladder (OAB) is a pervasive clinical problem involving alterations in both neurogenic and myogenic activity. While there has been some progress in understanding neurogenic inputs to OAB, the mechanisms controlling myogenic bladder activity are unclear. We report the involvement of myocardin (MYOCD) and microRNA‐1 (miR‐1) in the regulation of connexin 43 (GJA1), a major gap junction in bladder smooth muscle, and the collective role of these molecules during post‐natal bladder development. Wild‐type (WT) mouse bladders showed normal development from early post‐natal to adult including increases in bladder capacity and maintenance of normal sensitivity to cholinergic agents concurrent with down‐regulation of MYOCD and several smooth muscle cell (SMC) contractile genes. Myocardin heterozygous‐knockout mice exhibited reduced expression of Myocd mRNA and several SMC contractile genes concurrent with bladder SMC hypersensitivity that was mediated by gap junctions. In both cultured rat bladder SMC and in vivo bladders, MYOCD down‐regulated GJA1 expression through miR‐1 up‐regulation. Interestingly, adult myocardin heterozygous‐knockout mice showed normal increases in bladder and body weight but lower bladder capacity compared to WT mice. These results suggest that MYOCD down‐regulates GJA1 expression via miR‐1 up‐regulation, thereby contributing to maintenance of normal sensitivity and development of bladder capacity. J. Cell. Physiol. 228: 1819–1826, 2013. © 2013 Wiley Periodicals, Inc.     

NEW MATERIAL OF HSISOSUCHUS CHUNGKINGENSIS FROM SICHUAN, CHINA

AnewspecimenofHIitoIuchuschungkingensis,inc1udingthecraniumandmostofthep0stcranium,wascollectedbyone0fus(X.-M-Li)inearlyl960sfr0mthJurassicsandstoneoftheSangqia0area,Yongchuancounty,about60kmsouthwesIofChongqing.Thisspecimenpr0babIyrepresentsayoungadult0rasubadu1t,a5indicatedbyitssmal1size(lessthan2/3length0fthetype,basedontheskul]proportion)andopensuturesbetweenpalatalelements.lnthepresentpaperwewillredescribeH.chungkingensisindetailbasedonb0ththetypeandnewlydis-coveredspecimensinordertoc…     

Fascin promotes the invasion of pituitary adenoma through partial dependence on epithelial–mesenchymal transition

Journal of Molecular Histology - The aim of the present study was to investigate the role and potential regulatory mechanisms of fascin in the invasion and epithelial-to-mesenchymal transition of...     

IKK interacts with rictor and regulates mTORC2

mTORC2, the mammalian target of rapamycin complex 2 is activated by upstream growth factors, and performs two major functions, phosphorylation of AKT at the serine of 473 and cell cycle-dependent organization of actin cytoskeleton. However, the mechanisms through which mTORC2 is triggered by these signals remain unclear. We demonstrated, for the first time, that inhibitor of nuclear factor κ-B kinase (IKK) interacted with rictor and regulated mTORC2 activity. Not only endogenously, but ectopically expressed IKK α and IKK β physically interacted with rictor. An in vitro binding assay revealed that rictor interacted with IKKα and IKKβ from amino acids 999 to 1397. Moreover, chemical inhibition of IKK, knockdown of IKK by small interference RNA (siRNA), or ectopic expression of kinase-dead IKK (IKK KD) repressed phosphorylation of AKT (S473) in a variety of cell lines and decreased the kinase activity of mTORC2. In NIH 3 T3 cells, inhibition of IKK also reduced phosphorylation of protein kinase α (PKCα) (S657) and resulted in disorganization of actin cytoskeleton. Interestingly, the interaction between IKKα/β and rictor was increased, while the mTOR-rictor association was attenuated by inhibition of IKK. We identified a novel signaling mechanism for the regulation of mTORC2 by IKK: IKK interacted with rictor and regulated the function of mTORC2 including phosphorylation of AKT (S473) and organization of actin cytoskeleton. Inactivated IKK interacted with rictor and competed against mTOR, which resulted in a reduced mTORC2 level and a decrease in mTORC2 activity.     

小麦谷氨酰胺合成酶的原核表达特点

谷氨酰胺合成酶(GS)是植物氮同化的关键酶,为了研究小麦GS同工酶的结构及其表达特点,我们构建了小麦GS1、GSr、GSe、GS2和GS2前体GS2p的原核表达载体,并对表达条件进行了优化。结果表明,尽管小麦GS同工酶氨基酸序列同源性达70%–80%,蛋白质表达却各具特点。30℃诱导3 h后,GSr、GSe及GS2表达量达最大,诱导7 h后GS1表达量达最大,GS2p不表达,表达量依次为GS1(22%)GSr(15%)GS2(12%)GSe(5%);且GSe可溶性表达,GS1主要为可溶性表达,而GSr和GS2为包涵体。30℃诱导3 h,GS同工酶相对转录量为GSr(7.59)GS2(1.84)GS2p(1.66)GSe(1.46)GS1(1.00),酶蛋白质翻译水平与转录水平不一致。mRNA结构分析显示,GS同工酶翻译起始区稳定二级结构的自由能不同:GS1(14.4)GSr(17.2)GS2(22.6)GSe(25.4)GS2p(31.6),自由能越小,翻译起始区结构越不稳定,蛋白表达水平越高。GS1、GSr、GSe和GS2可溶性表达的最佳诱导条件不同,分别是30℃诱导5 h、16℃诱导15 h、37℃诱导5 h及25℃诱导7 h;可溶性表达量为GS1(20%)GSr(13%)GS2(10%)GSe(7%),酶活性为GS1GSeGS2,GSr无活性。可见,GS同工酶的基因序列决定了蛋白质在原核细胞中的表达量、状态及其活性。     

氮添加对戴云山黄山松林土壤有机氮解聚酶活性的影响及其调控因素

解聚作用是控制土壤有机氮矿化和氮素有效性供应的关键,然而氮沉降对亚热带森林土壤有机氮解聚作用的影响机制尚不明确。以福建戴云山黄山松林为研究对象,设置对照（CT）、低氮（LN）和高氮（HN）3个氮添加水平,进行为期2年的氮沉降模拟试验。通过分析土壤化学性质、微生物生物量和土壤8种有机氮解聚酶活性的变化,探究土壤有机氮解聚作用响应氮沉降的机理过程。结果表明：短期氮添加显著增加0-10 cm和10-20 cm土层矿质氮含量,并显著增加了10-20 cm土层微生物生物量碳（MBC）的含量。同时,0-10 cm土壤锰过氧化物酶活性随氮添加量增加而显著提高,HN处理下土壤漆酶活性显著高于LN和CT;10-20 cm土壤的酸性蛋白酶、碱性蛋白酶、中性蛋白酶和漆酶活性均随氮添加量增加而显著提高,但是谷氨酰胺酶活性变化相反。冗余分析表明两个土层有机氮解聚酶活性影响因素不同,土壤硝态氮（NO₃^--N）是0-10 cm土层有机氮解聚酶活性的主要影响因素,而10-20 cm土层有机氮解聚酶活性由NO₃^--N和MBC共同影响。综上所述,亚热带黄山松林土壤不同有机氮解聚酶对氮添加的响应不一致,主要受土壤NO₃^--N和MBC调节。该研究有助于拓宽土壤氮循环对氮沉降的响应机理,同时对维持土壤有效氮含量和提高黄山松生态系统生产力具有重要意义。