Synthesis,Characterization, In Vitro Cytotoxicity,and Apoptosis-Inducing Properties of Ruthenium(II) Complexes

Two new Ru(II) complexes, [Ru(bpy)₂(FAMP)](ClO₄)₂ 1 and 2, are synthesized and characterized by elemental analysis, electrospray mass spectrometry, and ¹H nuclear magnetic resonance. The in vitro cytotoxicities and apoptosis-inducing properties of these complexes are extensively studied. Complexes 1 and 2 exhibit potent antiproliferative activities against a panel of human cancer cell lines. The cell cycle analysis shows that complexes 1 and 2 exhibit effective cell growth inhibition by triggering G0/G1 phase arrest and inducing apoptosis by mitochondrial dysfunction. The in vitro DNA binding properties of the two complexes are investigated by different spectrophotometric methods and viscosity measurements.     

Antennal sensilla and their nerve innervation in first‐instar nymphs of Porphyrophora sophorae Arch. (Hemiptera: Coccomorpha: Margarodidae)

ABSTRACT Porphyrophora (Hemiptera: Coccomorpha: Margarodidae) is a genus of soil‐inhabiting scale insects. The antennal sensilla and their innervation in the first‐instar nymphs of Porphyrophora sophorae were studied using light microscopy and scanning and transmission electron microscopy to understand the function of these sensilla and determine the sensillar innervation feature on these small antennae. The results show that the six‐segmented antennae of these nymphs have 20–23 sensilla which can be morphologically classified into seven types, for example, one Böhm's bristle (Bb), one campaniform sensillum (Ca), one Johnston's organ (Jo), 13–16 aporous sensilla trichodea (St), two coeloconic sensilla (Co), one straight multiporous peg (Mp1), and one curvy multiporous peg (Mp2). According to their function, these sensilla can be categorized into three categories: mechanoreceptors, that is, Bb, Ca, Jo, and St; thermo/hygroreceptors, that is, Co only; and chemoreceptors, that is, Mp1 and Mp2. The dendrites that innervate the Mp1, Mp2, and Co sensilla combine to form a large nerve tract (NT1) in the antennal lumen. Because NT1 extends through and out of the antenna, the somata of these neurons are present in the lymph cavity of the insect's head. The dendrites that innervate the mechanoreceptors form another nerve tract (NT2). The somata of these neurons are located inside the scape and pedicel. J. Morphol. 277:1631–1647, 2016. © 2016 Wiley Periodicals, Inc.     

免疫细胞浸润在非小细胞肺癌诊断与预后中的应用

免疫细胞浸润对癌症的诊断与预后有着重要意义。文中收集TCGA数据库已收录的非小细胞肺癌肿瘤与正常组织基因表达数据,利用CIBERSORT工具得到22种免疫细胞占比来评估免疫细胞浸润情况。以22种免疫细胞占比为特征,用机器学习方法构建了非小细胞肺癌肿瘤与正常组织的分类模型,其中随机森林方法构建的模型分类效果AUC=0.987、敏感性0.98及特异性0.84。并且用随机森林方法构建的肺腺癌和肺鳞癌肿瘤组织分类模型效果AUC=0.827、敏感性0.75及特异性0.77。用LASSO回归筛选22种免疫细胞特征,保留8种强相关特征组成的免疫细胞评分结合临床特征构建了非小细胞肺癌预后模型。经评估及验证,预后模型C-index=0.71并且3年和5年的校准曲线拟合良好,可以对预后风险度进行准确预测。本研究基于免疫细胞浸润所构建的分类模型与预后模型,旨在对非小细胞肺癌的诊断与预后研究提供新的策略。     

Functional analysis of the dehydratase domains of a PUFA synthase from Thraustochytrium in Escherichia coli

Thraustochytrium sp. 26185, a unicellular marine protist, synthesizes docosahexaenoic acid, an omega-3 very long chain polyunsaturated fatty acid (VLC-PUFAs), by a polyunsaturated fatty acid (PUFA) synthase comprising three large subunits with multiple catalytic dehydratase (DH) domains critical for introducing double bonds at the specific position of fatty acids. To investigate functions of these DH domains, one DH domain from subunit-A and two DH domains from subunit-C of the PUFA synthase were dissected and expressed as stand-alone enzymes in Escherichia coli. The results showed that all these DH domains could complement the defective phenotype of a E. coli FabA temperature sensitive mutant, despite they have only modest sequence similarity with FabA, indicating they can function as 3-hydroxyacyl-ACP dehydratase for the biosynthesis of unsaturated fatty acids in E. coli. Site-directed mutagenesis analysis confirmed the authenticity of active site residues in these domains. In addition, overexpression of the three domains in a wild type E. coli strain resulted in the substantial alteration of fatty acid profiles including productions and ratio of unsaturated to saturated fatty acids. A combination of evidences from sequence comparison, functional expression, and mutagenesis analysis suggest that the DH domain from subunit-A is similar to DH domains from polyketide synthases, while the DH domains from subunit-C are more comparable to E. coli FabA in catalytic functions. Successful complementation and functional expression of the embedded DH domains from the PUFA synthase in E. coli is an important step towards for elucidating the molecular mechanism in the biosynthesis of VLC-PUFAs in Thraustochytrium.

     

miR24-2 accelerates progression of liver cancer cells by activating Pim1 through tri-methylation of Histone H3 on the ninth lysine

Several microRNAs are associated with carcinogenesis and tumour progression. Herein, our observations suggest both miR24-2 and Pim1 are up-regulated in human liver cancers, and miR24-2 accelerates growth of liver cancer cells in vitro and in vivo. Mechanistically, miR24-2 increases the expression of N6-adenosine-methyltransferase METTL3 and thereafter promotes the expression of miR6079 via RNA methylation modification. Furthermore, miR6079 targets JMJD2A and then increased the tri-methylation of histone H3 on the ninth lysine (H3K9me3). Therefore, miR24-2 inhibits JMJD2A by increasing miR6079 and then increases H3K9me3. Strikingly, miR24-2 increases the expression of Pim1 dependent on H3K9me3 and METTL3. Notably, our findings suggest that miR24-2 alters several related genes (pHistone H3, SUZ12, SUV39H1, Nanog, MEKK4, pTyr) and accelerates progression of liver cancer cells through Pim1 activation. In particular, Pim1 is required for the oncogenic action of miR24-2 in liver cancer. This study elucidates a novel mechanism for miR24-2 in liver cancer and suggests that miR24-2 may be used as novel therapeutic targets of liver cancer.