Triazole-fused sugars from nitroalkene-containing C-glycosides by a tandem 1,3-dipolar cycloaddition and intramolecular Michael addition

A synthetic method to triazole-fused sugars by treatment of nitroalkene-containing C-glycosides with sodium azide is described. Initial experiments conducted at room temperature gave only the 1,3-dipolar cycloaddition products. However, at elevated reaction temperature the tandem β-elimination/cycloaddition/Michael addition yielded 1,5-disubstituted triazole-fused sugars.     

CXCL16 Promotes Gastric Cancer Tumorigenesis via ADAM10-Dependent CXCL16/CXCR6 Axis and Activates Akt and MAPK Signaling Pathways

Abnormal expression of CXC motif chemokine ligand 16 (CXCL16) has been demonstrated to be associated with tumor progression and metastasis, served as a prognostic factor in many cancers, with higher relative expression behaving as a marker of tumor progression. However, its role and mechanisms underlying progression and metastasis of gastric cancer (GC) are yet to be elucidated. In our investigation, public datasets and human GC tissue samples were used to determine the CXCL16 expression levels. Our results revealed that CXCL16 was upregulated in GC. The high expression CXCL16 in GC was significantly associated with histologic poor differentiation and pTNM staging. And high CXCL16 was positively correlated with the poor survival of GC patients. Gain-and loss-of-function experiments were employed to investigate the biological role of CXCL16 in proliferation and migration both in vitro and in vivo. Mechanically, Gene set enrichment analysis (GSEA) revealed that the epithelial‑mesenchymal transition (EMT), Akt and MAPK signal pathway related genes were significantly enriched in the high CXCL16 group, which was confirmed by western blot. Moreover, overexpression CXCL16 promoted the disintegrin and metalloproteases (ADAM10) and the CXC motif chemokine receptor 6 (CXCR6) expression, which mediated the CXCL16/CXCR6 positive feedback loop in GC, with activating Akt and MAPK signaling pathways. Knocking down ADAM10 would interrupted the CXCL16/CXCR6 axis in the carcinogenesis and progression of GC. In conclusion, our findings offered insights into that CXCL16 promoted GC tumorigenesis by enhancing ADAM10-dependent CXCL16/CXCR6 axis activation.     

一株产纤维素酶细菌的筛选、鉴定及产酶条件优化

目的：筛选1株产纤维素酶的细菌。方法：通过对从腐烂朽木及其附近土壤中得到的样品进行富集培养、分离纯化得到16株纤维素分解菌,经刚果红染色鉴定和液体发酵培养后对其进行了菌种初步鉴定及产酶条件的初步优化。结果：获得1株纤维素酶分泌量较高的细菌LT3。结论：LT3为革兰氏阳性菌,菌体成杆状,经发酵优化培养后,较适产酶条件为甘蔗渣20g/L,pH7.0、30℃培养120h,CMC酶活为71.17U/mL,滤纸酶活为33.37U/mL。通过克隆其16S rDNA序列,对其进行系统进化分析,鉴定为蜡状芽孢杆菌。     

The multi-step process of human skin carcinogenesis: a role for p53, cyclin D1, hTERT, p16, and TSP-1

As proposed by Hanahan and Weinberg (2000. Cell 100, 57-70) carcinogenesis requires crucial events such as (i) genomic instability, (ii) cell cycle deregulation, (iii) induction of a telomere length maintenance mechanism, and (iv) an angiogenic switch. By comparing the expression of p53, cyclin D1, p16, hTERT, and TSP-1 in spontaneously regressing keratoacanthoma (KA) as a paradigm of early neoplasia, with malignant invasive cutaneous squamous cell carcinoma (SCC) as a paradigm of advanced tumour development, we are now able to assign the changes in the expression of these proteins to specific stages and allocate them to defined roles in the multi-step process of skin carcinogenesis. We show that mutational inactivation of the p53 gene, and with that the onset of genomic instability is the earliest event. Individual p53-positive cells are already seen in "normal" skin, and 3/5 actinic keratoses (AKs), 5/22 KAs, and 13/23 SCCs contain p53-positive patches. Cell cycle deregulation was indicated by the overexpression of the cell cycle regulator cyclin D1, as well as by the loss of the cell cycle inhibitor p16. Interestingly, overexpression of cyclin D1 - observed in 80% of KAs and SCCs, respectively - showed a cell cycle-independent function in HaCaT cell transplants on nude mice. Cyclin D1 overexpression was associated with a massive inflammatory response, finally leading to tissue destruction. Loss of the cell cycle inhibitor p16, on the other hand, correlated with SCCs. Thus, it is tempting to suggest that overexpression of cyclin D1 is an early change that in addition to growth stimulation leads to an altered epithelial-mesenchymal interaction, while functional p16 is able to control this deregulated growth and needs to be eliminated for malignant progression. Another requirement for uncontrolled growth is the inhibition of telomere erosion by up-regulating telomerase activity. As measured by hTERT protein expression, all of the KAs and SCCs studied were positive, with a similar distribution of the protein in both groups and an expression pattern resembling that of normal epidermis. Thus, telomerase may not need to be increased significantly in skin carcinomas. Finally, we show that the angiogenesis inhibitor TSP-1 is strongly expressed in most KAs, and mainly by the tumour cells, while in SCCs the generally weak expression is restricted to the tumour-stroma. Furthermore, we provide evidence that the loss of a copy of chromosome 15 is responsible for reduced TSP-1 expression and thereby this aberration contributes to tumour vascularisation (i.e. the angiogenic switch) required for malignant growth.     

IL-16及其前体对T细胞的调节作用

成熟的IL-16最初被称为淋巴细胞趋化因子,它是一种多效的细胞因子,能引发T淋巴细胞增殖,对于免疫反应和炎症具有调节作用。IL-16前体在胞质和胞核内均表达,是T细胞的生长阻遏剂。当抗原刺激时,胱天蛋白酶-3水解IL-16前体,大量的活性IL-16释放出来。该文主要介绍近年来IL-16及其前体对T细胞的调节作用的研究概况。     

The effects of intramolecular and intermolecular electrostatic repulsions on the stability and aggregation of NISTmAb revealed by HDX‐MS,DSC, and nanoDSF

The stability and aggregation of NIST monoclonal antibody (NISTmAb) were investigated by hydrogen/deuterium exchange mass spectrometry (HDX‐MS), differential scanning calorimetry (DSC), and nano‐differential scanning fluorimetry (nanoDSF). NISTmAb was prepared in eight formulations at four different pHs (pH 5, 6, 7, and 8) in the presence and absence of 150 mM NaCl and analyzed by the three methods. The HDX‐MS results showed that NISTmAb is more conformationally stable at a pH near its isoelectric point (pI) in the presence of NaCl than a pH far from its pI in the absence of NaCl. The stabilization effects were global and not localized. The midpoint temperature of protein thermal unfolding transition results also showed the C_H2 domain of the protein is more conformationally stable at a pH near its pI. On the other hand, the onset of aggregation temperature results showed that NISTmAb is less prone to aggregate at a pH far from its pI, particularly in the absence of NaCl. These seemingly contradicting results, higher conformational stability yet higher aggregation propensity near the pI than far away from the pI, can be explained by intramolecular and intermolecular electrostatic repulsion using Lumry‐Eyring model, which separates folding/unfolding equilibrium and aggregation event. The further a pH from the pI, the higher the net charge of the protein. The higher net charge leads to greater intramolecular and intermolecular electrostatic repulsions. The greater intramolecular electrostatic repulsion destabilizes the protein and the greater intermolecular electrostatic repulsion prevents aggregation of the protein molecules at pH far from the pI.     

某军校2014级与2010级新生16PF的比较分析

目的:比较某军校2014级与2010级本科新生以及14级新生内部的人格特点,为改善和提高军校的教育管理水平提供有效依据。方法:对2个年级新生进行16PF测试,对测查结果进行比较分析。结果:2014级新生在B、C、E、G、H、L、O、Q1、Q3、Q4、适应与焦虑性、内向与外向、心理健康因素、专业成就个性因素和新环境成长能力因素的得分与2010级新生得分有显著差异(P0.01)。从效果量角度分析,两级新生在A乐群性,适应与焦虑性,专业成就个性因素以及新环境成长能力因素等方面得分差异的效果量d大于0.2。2014级本科新生在A、B、C、E、I、L、M、O、Q1、Q2、Q3、Q4、感情用事与安详机警型、怯懦与果断型、心理健康因素、专业成就个性因素、创造能力个性因素以及新环境成长能力因素方面以及适应与焦虑性的得分与士官新生存在显著差异(P0.01)。从效果量角度分析,14级本科新生与士官新生在B,E,I,L,M,O,Q2,Q4,适应与焦虑性以及心理健康因素等方面得分差异的效果量d大于0.2,在数字搜索成绩,A,Q1,感情用事与安详机警型以及怯懦与果断型等方面得分差异的效果量d大于0.5。结论:14级不同身份学生存在各自特点,军对院校在对其教育管理上要注重方式方法,因材施教,合理引导,培养合格的军校学员,为我国培养优秀的军官。     

堇菜属组间的系统发育与亲缘关系：基于trnL-trnF、psbA-trnH、rpL16、ITS序列,细胞学以及形态学证据

以鼠鞭草（Hybanthus enneaspermus）、鳞隔堇（Scyphellandra pierrei）、雷诺木（Rinorea benga-lensis）作为外类群,对堇菜属（Viola）20个类群的trnL-trnF序列,17个类群的psbA-trnH、rpL16序列以及1个类群的nrDNAITS序列进行了测定,并从GenBank下载相应的序列,运用最大简约法以及贝叶斯推论法进行系统分析,构建系统发育树。结果表明：堇菜亚属（subgen. Viola）不是一个单系类群,并明确了堇菜属部分组间类群的亲缘关系。本文还结合形态与细胞学证据对堇菜属进行性状演化的推测。结果表明：1）直立茎较匍匐茎、莲座状茎（叶基生）原始;2）托叶边缘长流苏状与托叶1/2～3/4合生分别是鸟嘴柱头堇菜组（sect. Trigonocarpae）和合生托叶组（sect. Adnatae）演化路线的重要性状标志;3）花柱样式从柱头无喙演化至柱头有喙,并由柱头简单演化至柱头复杂,再趋向于柱头简化。     

Sec16 Determines the Size and Functioning of the Golgi in the Protist Parasite,Trypanosoma brucei

The Sec16 homologue in Trypanosoma brucei has been identified and characterized. TbSec16 colocalizes with COPII components at the single endoplasmic reticulum exit site (ERES), which is next to the single Golgi stack in the insect (procyclic) form of this organism. Depletion of TbSec16 reduces the size of the ERES and the Golgi, and slows growth and transport of a secretory marker to the cell surface; conversely, overexpression of TbSec16 increases the size of the ERES and Golgi but has no effect on growth or secretion. Together these data suggest that TbSec16 regulates the size of the ERES and Golgi and this size is set for optimal growth of the organism.      

拟诺卡氏菌16S rRNA,gyrB,sod和rpoB基因的系统发育分析

为了更好地了解拟诺卡氏菌属(Nocardiopsis)各物种间的系统发育关系,该属现有有效描述种的gyrB,sod和rpoB基因的部分序列被测定,结合16S rRNA基因,对拟诺卡氏菌属进行了系统发育重建。研究发现拟诺卡氏菌属gyrB,sod和rpoB基因的平均相似性分别为87.7%、87.3%和94.1%,而16S rRNA基因的平均相似性则达到96.65%,3个看家基因均比16S rRNA具有更高的分歧度。比较基于不同基因的系统树发现,由gyrB基因得到的系统树拓扑结构与16S rRNA得到的结构在亚群上基本一致。因此,gyrB基因在拟诺卡氏菌属的系统分类上比16S rRNA基因更具优越性。     

堇菜属组间的系统发育与亲缘关系：基于trnL-trnF、psbA trnH、rpL16、ITS序列，细胞学以及形态学证据

以鼠鞭草(Hybanthus enneaspermus)、鳞隔堇(Scyphellandra pierrei)、雷诺木(Rinorea bengalensis)作为外类群,对堇菜属(Viola)20个类群的trnL trnF序列,17个类群的psbA trnH、rpL16序列以及1个类群的nrDNA ITS序列进行了测定,并从GenBank下载相应的序列,运用最大简约法以及贝叶斯推论法进行系统分析,构建系统发育树。结果表明：堇菜亚属(subgen.Viola)不是一个单系类群,并明确了堇菜属部分组间类群的亲缘关系。本文还结合形态与细胞学证据对堇菜属进行性状演化的推测。结果表明：1）直立茎较匍匐茎、莲座状茎（叶基生）原始;2）托叶边缘长流苏状与托叶1/2~3/4合生分别是鸟嘴柱头堇菜组（sect.Trigonocarpae）和合生托叶组（sect.Adnatae）演化路线的重要性状标志;3）花柱样式从柱头无喙演化至柱头有喙,并由柱头简单演化至柱头复杂,再趋向于柱头简化。     

Fermented product of rice with Lactobacillus kefiranofaciens induces anti-aging effects and heat stress tolerance in nematodes via DAF-16

ABSTRACT

Rice kefiran is superior in functionality, has high concentration of mucilaginous polysaccharide, and low lipid content, compared to conventional kefiran. However, reports on its physiological functionality, especially studies on life expectancy and aging, in model organisms are rare. In this study, nematodes were used as model organisms that were fed rice kefiran, along with Escherichia coli OP50, as a result of which, the lifespan of nematodes was extended and age-related retardation of mobility was suppressed. It also increased the heat stress resistance in nematodes. Experiments using daf-16 deletion mutant revealed that rice kefiran functions via DAF-16. Thus, this study revealed the longevity, anti-aging and heat stress tolerance effects of rice kefiran in nematodes.     

Infrageneric Phylogeny of the Genus Viola (Violaceae) Based on trnL trnF,psbA trnH,rpL16, ITS Sequences,Cytological and Morphological Data

The sequences of chloroplast trnL trnF (20 species), psbA trnH(17 species), rpL16 (17 species) and ITS(one species) regions of the genus Viola were analyzed for phylogenetic relationships in the genus with the maximum parsimony (MP) and Bayesian inference (BI) analyses. Hybanthus enneaspermus, Scyphellandra pierrei and Rinorea bengalensis were used as outgroups. Additional sequences corresponding to the same DNA regions for the other taxa in Viola were downloaded from GenBank. The phylogenetic tree indicated that subgenViola is not monophyletic. Moreover, some intrageneric relationships in Viola were clarified. Combining sequences with morphological characters and chromosome number, the inference of morphological evolution in Viola was put forward. It was inferred that: 1) erect stems might well be more primitive than stolons and rosettes; 2) long fimbriate margin in stipules and stipules with 1/2-3/4 adnate part might represent distinctive evolutionary trends of sectTrigonocarpae and of sectAdnatae respectively; 3) species with stigmatic beaks might have been derived from ancestors without beaks; simple structure stigmas transformed into complex structure stigmas, and then transformed back to simple stucture patterns.     

铜绿假单胞菌氨基糖苷16SrRNA甲基化酶基因分析

目的调查215株湖州地区临床分离铜绿假单胞菌对氨基糖苷类抗生素的耐药性和16S rRNA甲基化酶基因分布情况。方法收集2011年1月至2012年12月湖州地区临床分离铜绿假单胞菌215株,琼脂稀释法测定5种氨基糖苷类抗菌药物（庆大霉素、阿米卡星、妥布霉素、伊帕米星、奈替米星）的MIC值;PCR检测armA、rmtA、rmtB、rmtC、rmtD和npmA六种氨基糖苷类16S rRN甲基化酶基因,序列分析明确基因型。测定产16S rRNA甲基化酶菌株对常见抗菌的敏感性,并检测碳青霉烯耐药株产碳青霉烯酶情况。结果铜绿假单胞菌对异帕米星敏感率最高为81.4%,对5种氨基糖苷类抗生素全部耐药的22株菌株中,17株检出armA基因;未发现其他16S rRNA甲基化酶基因阳性菌株。17株armA阳性菌株对碳青霉烯类抗生素耐药5株（耐药率为29.4%）,对头孢他啶、头孢吡肟、哌拉西林/他唑巴坦、环丙沙星耐药率均超过40%。5株碳青霉烯耐药菌株中检测到2株产VIM-2型金属碳青霉烯酶。结论铜绿假单胞菌对氨基糖苷类抗生素耐药率高,检测到16S rRNA甲基化酶基因armA。产16S rRNA甲基化酶铜绿假单胞菌耐药性强,部分菌株同时产金属碳青霉烯酶,给临床抗感染治疗及院内感染控制带来挑战。     

The C-terminal tail extension of myosin 16 acts as a molten globule,including intrinsically disordered regions,and interacts with the N-terminal ankyrin

The lesser-known unconventional myosin 16 protein is essential in proper neuronal functioning and has been implicated in cell cycle regulation. Its longer Myo16b isoform contains a C-terminal tail extension (Myo16Tail), which has been shown to play a role in the neuronal phosphoinositide 3-kinase signaling pathway. Myo16Tail mediates the actin cytoskeleton remodeling, downregulates the actin dynamics at the postsynaptic site of dendritic spines, and is involved in the organization of the presynaptic axon terminals. However, the functional and structural features of this C-terminal tail extension are not well known. Here, we report the purification and biophysical characterization of the Myo16Tail by bioinformatics, fluorescence spectroscopy, and CD. Our results revealed that the Myo16Tail is functionally active and interacts with the N-terminal ankyrin domain of myosin 16, suggesting an intramolecular binding between the C and N termini of Myo16 as an autoregulatory mechanism involving backfolding of the motor domain. In addition, the Myo16Tail possesses high structural flexibility and a solvent-exposed hydrophobic core, indicating the largely unstructured, intrinsically disordered nature of this protein region. Some secondary structure elements were also observed, indicating that the Myo16Tail likely adopts a molten globule–like structure. These structural features imply that the Myo16Tail may function as a flexible display site particularly relevant in post-translational modifications, regulatory functions such as backfolding, and phosphoinositide 3-kinase signaling.     

Improved synthesis of 17β-hydroxy-16α-iodo-wortmannin, 17β-hydroxy-16α-iodoPX866, and the [131I] analogue as useful PET tracers for PI3-kinase

An improved method for the synthesis of 17β-hydroxy-16α-iodo-wortmannin along with the first synthesis of 17β-hydroxy-16α-iodoPX866 and [¹³¹I] radiolabeled 17β-hydroxy-16α-[¹³¹I]iodo-wortmannin, as potential PET tracers for PI3K was also described. The differences between wortmannin and its iodo analogue were compared by covalently docking each structure to L833 in PI3K.     

16SrDNA同源性所揭示的双歧杆菌与有关细菌的亲缘关系

本研究测定了低ＧＣ含量的双歧杆菌（Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍ ｉｎｏｐｉｎａｔｕｍ）和新种Ｂ。ｔｈｅｒｍｏｃｉ．ｄｏｐｈｉｌｕｍ的１６ＳｒＤＮＡ全序列,在同另外１９个双歧杆菌及８个相关细菌的１６ＳｒＮＡ同源笥分析的基础上构建了系统发育树。结果表明：除低ＧＣ含蜈的Ｂ．ｉｎｏｐｉｎａｔｕｍ外,所有双杆菌的种在１６ＳｒＤＮＡ序列相似性≥８２％的水平上聚类为一个簇群。尽管Ｂ．ｉｎｏｐｉｎａｉｕｍ与其它     

Phylogenetic Analysis of the Genus Bifidobacterium and Related Genera Based on 16S rDNA Sequences

The 16S rRNA gene sequences were determined for type strains of 21 Bifidobacterium species. A phylogenetic tree was constructed using the determined sequences and sequences from DNA databases, which contain the sequences of 11 type strains of Bifidobacterium species and 11 strains of related genera. All species of the genus Bifidobacterium and Gardnerella vaginalis ATCC 14018 belonged to a cluster phylogenetically distinct from the other genera. The cluster was divided into two subclusters: subcluster 1 composed of most species of Bifidobacterium and G. vaginalis, and subcluster 2 consisting of two species, B. denticolens and B. inopinatum; both of which were isolated from human dental caries. In the genus Bifidobacterium, four groups of species are known to be moderately to highly related by DNA-DNA hybridization. The four groups of species exhibited more than 99% similarity among their 16S rDNA sequences within each group. These results indicated that species with around 99% or more similarity in their 16S rDNA sequences should be confirmed for species identities.