Deciphering binding affinity,energetics, and base specificity of plant alkaloid Harmane with AT and GC hairpin duplex DNA

Insights into binding efficacy and thermodynamic aspects of small molecules are important for rational drug designing and development. Here, the interaction of Harmane (Har), a very important bioactive indole alkaloid, with AT and GC hairpin duplex−DNAs has been reported using various biophysical tools. Detailed molecular mechanism with special emphasis on binding nature, base specificity, and thermodynamics have been elucidated via probing nucleic acids with varying base compositions. Har bound to both the DNA strands exhibited hypochromic effect in absorbance whereas bathochromic and hypochromic effects in fluorescence spectra. The binding constants estimated were in the order of 10⁵ M⁻¹ (higher for GC sequence compared with AT) with 1:1 stoichiometry. Noncooperative binding mode has been observed via intercalation in both the cases. The thermodynamic profile was obtained from temperature-dependent fluorescence experiments. Both Har–AT and Har–GC complexations were exothermic in nature associated with positive entropy and negative enthalpy changes. Salt-dependent studies revealed that the binding interaction was governed by nonpolyelectrolytic and hydrophobic interaction forces. The ligand-induced structural perturbation of the DNA structures was evident from the circular dichroism data. Molecular modelling data indicated towards the involvement of hydrophobic forces and hydrogen bonding.     

康力欣胶囊联合FOLFOX4方案对中晚期胃癌患者免疫功能、生活质量和血清肿瘤标志物的影响

目的:观察康力欣胶囊联合奥沙利铂、亚叶酸钙、5-氟尿嘧啶(FOLFOX4)方案对中晚期胃癌患者免疫功能、生活质量和血清肿瘤标志物的影响。方法:前瞻性选取2018年7月~2021年1月期间来我院接受治疗的胃癌患者80例,根据抽签分为对照组和观察组两组,各为40例。对照组接受FOLFOX4方案治疗,观察组接受康力欣胶囊联合FOLFOX4方案治疗,以3周为1个疗程,治疗2个疗程。观察两组治疗2个疗程后的疗效、生活质量以及治疗期间不良反应状况,对比两组治疗前、治疗2个疗程后的肿瘤标志物和免疫功能指标变化情况。结果:观察组治疗2个疗程后的总有效率高于对照组(P<0.05)。治疗2个疗程后,观察组的肿瘤相关物质群(TSGF)、糖类抗原-199(CA-199)、癌胚抗原(CEA)低于对照组(P<0.05)。治疗2个疗程后,观察组的CD3;、CD4;、自然杀伤细胞(NK)、CD4;/CD8;高于对照组,CD8;低于对照组(P<0.05)。治疗2个疗程后,观察组的生活质量总改善率、卡氏评分(KPS)评分高于对照组(P<0.05)。两组不良反应发生率组间对比无统计学差异(P>0.05)。结论:中晚期胃癌患者采用康力欣胶囊联合FOLFOX4方案治疗,在阻止疾病进展、改善生活质量、提高免疫功能方面均效果确切,优于单纯化疗效果。     

中国药用当归属植物研究进展及质量标志物的预测分析*

中国药用当归属植物资源丰富,包括当归、白芷、独活等34种药材,临床使用广且药用价值极高,富含香豆素类、多糖类、有机酸类、苯酚类、挥发油类等成分,除补血活血、调经止痛、润肠通便的传统功效,还具有抗氧化、抗衰老、保护神经元、抗肿瘤等药理作用,应用前景广阔。综述了中国药用当归属植物的分布及其药用功效、化学成分和药理作用的研究进展,为当归属植物的开发利用提供参考。并且根据质量标志物(Q-marker)概念,把影响中国药用当归属植物质量的因素概括为当归属药用植物品种、植物产区、化学成分、炮制方法,以及传统功效和网络药理学的预测分析等。从不同方面对中国药用当归属植物进行分析和论述,为建立和完善中国药用当归属植物的质量控制及评价方法提供参考。     

Formulation matters! A spectroscopic and molecular dynamics investigation on the peptide CIGB552 as itself and in its therapeutical formulation

Synthetic therapeutic peptides (STP) are intensively studied as new-generation drugs, characterized by high purity, biocompatibility, selectivity and stereochemical control. However, most of the studies are focussed on the bioactivity of STP without considering how the formulation actually used for therapy administration could alter the physico-chemical properties of the active principle. The aggregation properties of a 20-mer STP (Ac-His-Ala-Arg-Ile-Lys-D-Pro-Thr-Phe-Arg-Arg-D-Leu-Lys-Trp-Lys-Tyr-Lys-Gly-Lys-Phe-Trp-NH₂), showing antitumor activity, were investigated by optical spectroscopy and atomic force microscopy imaging, as itself (CIGB552) and in its therapeutic formulation (CIGB552TF). It has found that the therapeutic formulation deeply affects the aggregation properties of the investigated peptide and the morphology of the aggregates formed on mica by deposition of CIGB552 and CIGB552TF millimolar solutions. Molecular dynamics simulations studied the first steps of CIGB552 aggregation under physiological ionic strength conditions (NaCl 150 mM), showing that peptide oligomers, from dimers to tetramers, are preferentially formed in this environment. Interestingly, cell viability assays performed on H-460 cell lines indicate a major antiproliferative activity of the peptide in its therapeutic formulation with respect to the peptide aqueous solution.     

The complete mitochondrial genome of Tuta absoluta (Lepidoptera: Gelechiidae) and genetic variation in two newly invaded populations in China

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating invasive pest worldwide, causing severe damage to tomatoes. Recently, it has been recorded in the northwestern and southwestern parts of China. Here, the mitogenomes and genetic variation of two newly invaded T. absoluta populations in Xinjiang and Yunnan, were determined. The results showed that the complete mitogenome size of T. absoluta is 15298 bp for the individual from Xinjiang and 15296 bp for the individual from Yunnan, which were both longer than the reported mitogenome from Spain (15290 bp). The mitogenome sequences of individuals collected from three locations showed high levels of sequence similarity, except for 8 polymorphic sites, which were in genes cox2 (1 site), cox3 (2 sites), cob (1 site), atp6 (1 site), nad1 (2 sites) and nad5 (1 site). Tuta absoluta mitogenomes share many features with other 6 Gelechiidae mitogenomes, except for several differences in the start and stop codons of protein-coding genes and the length of intergenic spacers. Seven partial mitochondrial genes (cox1, cox2, cox3, atp6, cob, nad1, and nad5) were used for genetic variation analysis, and significant population differentiation was found between the two populations based on cox2, atp6, nad1, and nad5. The complete mitogenomes and sensitive mitochondrial gene markers reported here provide useful data for further population genetics study of this pest.     

基于分子标记的入侵陕西草地贪夜蛾种群生物型分析

草地贪夜蛾Spodoptera frugiperda J. E. Smith是一种世界性入侵害虫,为了明确入侵陕西的草地贪夜蛾的种群生物型并了解其发生及扩散规律,本研究对采集自陕西省8个地市180个草地贪夜蛾样本分别进行了基于COI和Tpi分子标记的生物型鉴定。分析发现入侵陕西的草地贪夜蛾84%是水稻型母本与玉米型父本杂交形成的杂合玉米型草地贪夜蛾。COI分子标记的结果显示,样本中水稻型占比为84.44%,玉米型为15.56%;基于Tpi基因片段的结果表明除商洛样本SL-3外,其它样本均为玉米型。值得注意的是,SL-3与非洲特异型序列同源性达100%,此非洲特异单倍型为陕西省内首次、国内第2次出现。本研究为草地贪夜蛾的迁飞扩散规律及早期预警提供了新的理论基础。     

Autophagy protects against redox-active trace metal-induced cell death in rabbit synovial fibroblasts through Toll-like receptor 4 activation

Reactive oxygen species (ROS) are implicated to play a role in initiating rheumatoid arthritis (RA) pathogenesis. We have investigated the mechanism(s) by which essential redox-active trace metals (RATM) may induce cell proliferation and cell death in rabbit synovial fibroblasts. These fibroblast-like synovial (FLS) cells, which express Toll-like receptor 4 (TLR4), were used as a model system that plays a role in potentially initiating RA through oxidative stress. Potassium peroxychromate (PPC, [Cr⁵⁺]), ferrous chloride (FeCl₂, [Fe²⁺]), and cuprous chloride (CuCl, [Cu⁺]) in the indicated valency states were used as exogenous pro-oxidants that can induce oxidative stress through TLR4 coupled activation that also causes HMGB1 release. We measured the proliferation index (PI) of FLS, and examined the effect of RATM oxidants on apoptosis and autophagy by fluorescence cell-sorting flow cytometry (FC). Cell cycle was analysed by FC and autophagy-related protein expression levels were measured by western blot. Our data showed that as RATM as prooxidants increased intracellular ROS (iROS) that can induce oxidative stress. Whereas iROS increased PI in FLS, these reactive species also protected cells against apoptosis by inducing autophagy. Our results indicate that ROS/TLR4-coupled activation may contribute to the pathogenesis of RA in FLS by induction of autophagy. The signalling pathway by which inflammation and its tissue destructive sequel may occur in RA underlies the need for developing therapeutic agents that can inhibit release of tissue-damaging high mobility group box 1 (HMGB1), cytokines, and possess both trace metal chelating capacity and oxidant scavenging properties in a directed combinatorial therapy for RA.     

The PDZ-GEF Gef26 regulates synapse development and function via FasII and Rap1 at the Drosophila neuromuscular junction

Guanine nucleotide exchange factors (GEFs) are essential for small G proteins to activate their downstream signaling pathways, which are involved in morphogenesis, cell adhesion, and migration. Mutants of Gef26, a PDZ-GEF (PDZ domain-containing guanine nucleotide exchange factor) in Drosophila, exhibit strong defects in wings, eyes, and the reproductive and nervous systems. However, the precise roles of Gef26 in development remain unclear. In the present study, we analyzed the role of Gef26 in synaptic development and function. We identified significant decreases in bouton number and branch length at larval neuromuscular junctions (NMJs) in Gef26 mutants, and these defects were fully rescued by restoring Gef26 expression, indicating that Gef26 plays an important role in NMJ morphogenesis. In addition to the observed defects in NMJ morphology, electrophysiological analyses revealed functional defects at NMJs, and locomotor deficiency appeared in Gef26 mutant larvae. Furthermore, Gef26 regulated NMJ morphogenesis by regulating the level of synaptic Fasciclin II (FasII), a well-studied cell adhesion molecule that functions in NMJ development and remodeling. Finally, our data demonstrate that Gef26-specific small G protein Rap1 worked downstream of Gef26 to regulate the level of FasII at NMJs, possibly through a βPS integrin-mediated signaling pathway. Taken together, our findings define a novel role of Gef26 in regulating NMJ development and function.     

Ovarian cancer stem cell: A potential therapeutic target for overcoming multidrug resistance

The cancer stem cell (CSC) model encompasses an advantageous paradigm that in recent decades provides a better elucidation for many important biological aspects of cancer initiation, progression, metastasis, and, more important, development of multidrug resistance (MDR). Such several other hematological malignancies and solid tumors and the identification and isolation of ovarian cancer stem cells (OV-CSCs) show that ovarian cancer also follows this hierarchical model. Gaining a better insight into CSC-mediated resistance holds promise for improving current ovarian cancer therapies and prolonging the survival of recurrent ovarian cancer patients in the future. Therefore, in this review, we will discuss some important mechanisms by which CSCs can escape chemotherapy, and then review the recent and growing body of evidence that supports the contribution of CSCs to MDR in ovarian cancer.