Advance in research of microRNA in Caenorhabditis elegans

microRNA (miRNA) is a family of small, non‐coding RNA first discovered as an important regulator of development in Caenorhabditis elegans (C. elegans). Numerous miRNAs have been found in C. elegans, and some of them are well conserved in many organisms. Though, the biologic function of miRNAs in C. elegans was largely unknown, more and more studies support the idea that miRNA is an important molecular for C. elegans. In this review, we revisit the research progress of miRNAs in C. elegans related with development, aging, cancer, and neurodegenerative diseases and compared the function of miRNAs between C. elegans and human. J. Cell. Biochem. 114: 994–1000, 2013. © 2012 Wiley Periodicals, Inc.     

Rootstocks improve cucumber photosynthesis through nitrogen metabolism regulation under salt stress

We examined the growth, photosynthetic parameters, initial and total ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity, the relative expression of rbcL, rbcS, and rca gene, and nitrogen metabolism of cucumber (Cucumis sativus L. cv. Jinchun No.2, CS) plants grafted onto figleaf gourd (Cucurbita ficifolia Bouché, CF) and pumpkin (Cucurbita moschata Duch. cv. Chaojiquanwang, CM) rootstocks. Growth inhibition under salt stress (90 mM NaCl) was characterized by the irreversible inhibition of CO₂ assimilation in the cucumber plants grafted onto cucumber rootstocks (CS/CS). In contrast, this effect was significantly alleviated by grafting the cucumber plants onto the CF and CM roots (CS/CF, CS/CM). Under NaCl stress, the CS/CF and CS/CM plants exhibited higher photosynthetic activity, higher initial and total Rubisco activity, and higher Rubisco-related gene expression than the CS/CS plants. Salinity resulted in a lesser increase in nitrate content and decrease in free amino acid content in the CS/CF and the CS/CM plants compared with the CS/CS plants. Accordingly, the activity of nitrate reductase, glutamine synthetase, and glutamate synthase decreased significantly, especially in the CS/CS plants. These results suggest that grafting cucumber plants onto salt-tolerant rootstocks enhances Rubisco activity and the expression of Rubisco-related genes by effectively accelerating nitrate transformation into amino acids under NaCl stress, thereby improving the photosynthetic performance of cucumber leaves.     

Native mass spectrometry for understanding dynamic protein complex

Biomolecules have evolved to perform specific and sophisticated activities in a highly coordinated manner organizing into multi-component complexes consisting of proteins, nucleic acids, cofactors or ligands. Understanding such complexes represents a task in earnest for modern bioscience. Traditional structural techniques when extrapolating to macromolecules of ever increasing sizes are confronted with limitations posed by the difficulty in enrichment, solubility, stability as well as lack of homogeneity of these complexes. Alternative approaches are therefore prompted to bridge the gap, one of which is native mass spectrometry. Here we demonstrate the strength of native mass spectrometry, used alone or in combination with other biophysical methods such as analytical ultracentrifugation, small-angle neutron scattering, and small-angle X-ray scattering etc., in addressing dynamic aspects of protein complexes including structural reorganization, subunit exchange, as well as the assembly/disassembly processes in solution that are dictated by transient non-covalent interactions. We review recent studies from our laboratories and others applying native mass spectrometry to both soluble and membrane-embedded assemblies. This article is part of a Special Issue entitled “Biophysical Exploration of Dynamical Ordering of Biomolecular Systems” edited by Dr. Koichi Kato.     

Toll-like receptor 2 activation implicated in oral squamous cell carcinoma development

Recent studies have revealed that Toll-like receptors (TLRs) are highly expressed and activated in many types of cancer. Physiologically, TLR2 recognizes bacteria and other microorganisms in the oral cavity; however, the role of TLR2 in oral squamous cell carcinoma (OSCC) is unclear. In this study, we demonstrated that TLR2 is highly expressed in OSCC in comparison with adjacent non-malignant tissue. TLR2 was also expressed in OSCC-derived cell lines, and its expression was activated by ligands derived from bacteria and mycoplasma. Furthermore, to elucidate the mechanism of OSCC progression via TLR2 signal transduction, we focused on microRNAs (miRNAs) that are induced by TLR2 activation. Interestingly, ligand activation of TLR2 induced the expression of miR-146a and we found that downregulation of caspase recruitment domain–containing protein 10 (CARD10) mRNA in OSCC-derived cell lines. Moreover, knockdown of CARD10 induced resistance to cisplatin-induced apoptosis in OSCC cells. These findings suggest that the activation of TLR2 by bacterial components can enhance the progression of OSCC and may be implicated in acquired resistance to cisplatin-induced apoptosis through regulation of the miR-146a pathway.     

紫穗槐幼苗根系生理特性和解剖结构对PEG-6000模拟干旱的响应

采用PEG-6000模拟干旱胁迫处理,测定了紫穗槐幼苗根系的可溶性糖、可溶性蛋白质、丙二醛、游离脯氨酸含量及SOD、POD酶活性变化以及解剖结构特征,旨在比较不同干旱程度对紫穗槐幼苗根系生理指标、内部解剖结构的影响,探索紫穗槐幼苗对水分胁迫的适应能力,揭示紫穗槐幼苗根系对土壤水分胁迫的响应和调控机制。结果表明:丙二醛含量变化显示当PEG-6000溶液浓度超过50g/L以后,紫穗槐幼苗根的膜系统开始受到损伤,并在PEG-6000溶液浓度达到250g/L受损程度显著增强,达到了对照的1.6倍,同时启动渗透调节作用(游离脯氨酸含量显著增加),达到了对照的3.8倍,在PEG-6000溶液浓度低于200g/L时,紫穗槐幼苗根系中至少没有启动以游离脯氨酸为主的渗透调节过程。可溶性糖和可溶性蛋白质含量及SOD、POD酶活性的变化印证了胞内发生的生理代谢变化,在PEG-6000溶液浓度为200g/L时,可溶性糖含量仅为0.121mg/g,达到最低点,随后上升,当PEG-6000溶液浓度进一步增加到250g/L时,紫穗槐幼苗根系中的可溶性糖含量则迅速回升到0.64mg/g,为对照组的63.37%。可溶性蛋白质含量在低浓度PEG-6000溶液(50g/L)处理下即有明显反应,下降到对照的61.5%,随后呈波动性变化。SOD和POD活性对PEG-6000模拟干旱胁迫的响应规律类似,均对PEG-6000模拟干旱胁迫处理迅速响应且活性增加。当PEG-6000溶液浓度达到50g/L至100g/L时,抗氧化酶的合成量最高,而后活性下降。60d的PEG-6000模拟干旱胁迫处理影响了紫穗槐幼苗根系的生长发育,随着PEG-6000溶液浓度增加,维管柱的直径变大,木质部厚度增大,导管直径变小、但导管密度增加,当PEG-6000溶液浓度达到250g/L时,导管密度比对照组增加了41.3%,木质部厚度比对照组增加了91.5%。以上结果表明,PEG-6000模拟干旱胁迫处理下,不同胁迫程度紫穗槐内部生理和根系解剖结构变化不同,通过改变自身生理代谢和根系内部解剖结构,以适应土壤水分胁迫的逆境条件,来满足自身生长和发育的需求平衡。     

Restoration of miR-98 relieves the inhibitory effect of nicotine on the differentiation of P19 cells into cardiomyocytes

Objective

To study whether miR-98 participates in the effects of nicotine on myocardial differentiation.

Results

By western blot, MTT and flow cytometry assays, we found that nicotine suppresses P19 cell differentiation into cardiomyocytes and apoptosis, and promotes proliferation, while restoration of miR-98 relieves the inhibitory effect of nicotine on the P19 cell differentiation. By target prediction analysis and luciferase reporter assay, we observed that miR-98 inhibits the protein expression of Wnt1 by directly acting on the 3′-UTR of Wnt1 mRNA. We assumed that the effect of miR-98 on Wnt1 might alter the activity of the Wnt1/β-catenin signaling pathway and be associated with myocardial differentiation. In summary, nicotine restrains differentiation of P19 cells into cardiomyocytes and decreases the level of miR-98.

Conclusions

Restoration of miR-98 relieves the inhibitory effect of nicotine on differentiation of P19 cells via targeting the 3′-UTR of Wnt1, which offers novel insights into our understanding of underlying molecular mechanisms of congenital heart defects.

     

Alpha-melanocyte-stimulating hormone mediates melanin-concentrating hormone-induced anorexigenic action in goldfish

In goldfish, intracerebroventricular (ICV) administration of melanin-concentrating hormone (MCH) inhibits feeding behavior, and fasting decreases hypothalamic MCH-like immunoreactivity. However, while MCH acts as an anorexigenic factor in goldfish, in rodents MCH has an orexigenic effect. Therefore, we examined the involvement of two anorexigenic neuropeptides, alpha-melanocyte-stimulating hormone (alpha-MSH) and corticotropin-releasing hormone (CRH), in the anorexigenic action of MCH in goldfish, using an alpha-MSH receptor antagonist, HS024, and a CRH receptor antagonist, alpha-helical CRH((9-41)). ICV injection of HS024, but not alpha-helical CRH((9-41)), suppressed MCH-induced anorexigenic action for a 60-min observation period. We then examined, using a real-time PCR method, whether MCH affects the levels of mRNAs encoding various orexigenic neuropeptides, including neuropeptide Y (NPY), orexin, ghrelin and Agouti-related peptide (AgRP), in the goldfish diencephalon. ICV administration of MCH at a dose sufficient to inhibit food consumption decreased the expression of mRNAs for NPY and ghrelin, but not for orexin and AgRP. These results indicate that the anorexigenic action of MCH in the goldfish brain is mediated by the alpha-MSH signaling pathway and is accompanied by inhibition of NPY and ghrelin synthesis.     

烟草废料不同方法浸提烟碱的研究

对利用烟草废料浸提烟碱进行了研究,探讨了浸提温度、时间、液固比、浸提溶剂等因素对浸提的影响。实验结果表明,使用乙醇作为浸提溶剂,在78℃,液固比5：1,浸提时间6．5h的条件下,浸提液中烟碱质量浓度为19．3mg/mL。用微波处理时,可以用水作为浸提溶剂,浸提液中烟碱质量浓度为11．1mg/mL。