Identification of the glucosamine kinase in the chitinolytic pathway of Thermococcus kodakarensis

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植物多酚通过MAPK信号通路调控肿瘤作用机制研究进展

肿瘤细胞的生物学过程(增殖、分化、侵袭、迁移)是癌症发生的标志,丝裂原活化蛋白激酶(mitogenactivated protein kinase,MAPK)被证实是调节肿瘤细胞生物学过程的至关重要的信号通路。大量研究表明,天然植物多酚,如茶多酚、白藜芦醇、花青素等对肿瘤细胞有显著的调节作用,植物多酚介导的MAPK通路对肿瘤细胞的调节已经引起国内外学者的广泛兴趣。文章综述了植物多酚通过MAPK信号通路对肿瘤细胞的调控作用,分析了不同多酚对MAPK信号通路的4条途径(p38、ERK-1/2、ERK5、JNK)的响应机制,旨在为明确植物多酚的抗肿瘤活性及分子机制、开发抗肿瘤保健食品或药物提供参考。     

Gene regulation in response to protein disulphide isomerase deficiency

We have examined the activities of promoters of a number of yeast genes encoding resident endoplasmic reticulum proteins, and found increased expression in a strain with severe protein disulphide isomerase deficiency. Serial deletion in the promoter of the MPD1 gene, which encodes a PDI1-homologue, revealed a cis-acting element responding to deficiency of protein disulphide isomerase activity (designated CERP). The presence of the sequence element is necessary and sufficient for the upregulation in response to disulphide isomerase deficiency, as measured by a minimal promoter containing the CERP element. The sequence (GACACG) does not resemble the unfolded protein response element. It is present in the upstream regions of the MPD1, MPD2, KAR2, PDI1 and ERO1 genes.     

辣木异硫氰酸酯通过活化AMPK抑制3T3-L1脂肪细胞脂质积累

探究辣木异硫氰酸酯-4-[(α-L-rhamnosyloxy) benzyl] Isothiocyanates（MIC-1）抑制3T3-L1脂肪细胞脂质积累的作用及可能的调控机制。体外诱导3T3-L1前脂肪细胞分化为成熟脂肪细胞,用MIC-1干预48 h后检测细胞脂质积累情况,甘油三酯（TG）、甘油（Gly）和游离脂肪酸（FFA）含量;qRT-PCR检测脂代谢相关基因的表达;Western blot法测定腺苷酸活化蛋白激酶（AMPK）蛋白磷酸化水平和氧化物酶体增殖激活受体γ（PPARγ）蛋白表达水平。结果表明,MIC-1对3T3-L1前脂肪细胞存活率无影响;与对照组相比,MIC-1可降低脂肪细胞内脂滴分布及细胞着色程度,降低细胞内TG含量,减少FFA及甘油的溢出。MIC-1处理浓度达4 mol/L时,TG和FFA浓度分别下降64.00%和75.00%。同时,显著下调细胞中PPARγ（46.00%）、硬脂酰辅酶A去饱和酶1（SCD1）（62.00%）的mRNA表达水平;显著上调AMPK蛋白磷酸化水平（64.47%）和下调PPARγ（52.10%）蛋白表达水平。以上结果表明,MIC-1通过促进TG分解和抑制TG的合成,从而抑制脂质积累,其机制可能与AMPK的活化有关。     

p-辛弗林通过激活AMPK-FoxO1信号通路抑制肝细胞葡萄糖生成

目的：探讨p-辛弗林对肝细胞葡萄糖生成的影响及其作用机制。方法：采用MTS法检测p-辛弗林对HepG2肝细胞株的细胞活力的影响,确定受试物的作用浓度后,比色法测定葡萄糖的生成、葡萄糖-6-磷酸酶（glucose-6-phosphatase,G6Pase）和磷酸烯醇丙酮酸羧基激酶（phosphoenolpyruvate carboxykinase,PEPCK）活力,Western blot蛋白印迹法检测腺苷酸活化蛋白激酶（adenosine 5’-monophosphate-activated protein kinase,AMPK）、磷酸化AMPK（p-AMPK）、乙酰辅酶A羧化酶（acetyl coenzyme A synthetase,ACC）、磷酸化ACC（p-ACC）、叉头框转录因子1（forkhead box class O1,FoxO1）以及磷酸化FoxO1（p-FoxO1）的表达;利用AMPK选择性抑制剂Compound C和AMPK siRNA作用HepG2肝细胞株后,检测p-辛弗林对HepG2肝细胞株葡萄糖的生成、G6Pase和PEPCK活力的影响。结果：p-辛弗林能剂量依赖性地抑制肝细胞葡萄糖的生成,激活AMPK信号通路,促进p-AMPK、p-ACC和p-FoxO1水平增加,极显著抑制G6Pase和PEPCK的活性（P＜0.01）。p-辛弗林的这些影响部分地被Compound C和AMPK siRNA所抑制（P＜0.01）。结论：p-辛弗林能够通过激活AMPK-FoxO1信号通路抑制肝细胞葡萄糖生成。     

中波紫外线致皮肤光老化机制及茶叶抗光老化作用研究进展

过量的紫外线照射会引起皮肤的光老化,其中以中波紫外线生物学效应最为明显。皮肤的光老化会导致皮肤临床上和组织学上的多种病变,包括胶原蛋白减少、皮肤变硬、皮层变薄、色素沉积等,并会引起日光性皮炎等多种皮肤相关疾病。中波紫外线诱导皮肤光老化的原因很多,其中最重要的原因是丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)通路被激活,引起金属基质蛋白酶(matrix metalloproteinase,MMPs)分泌量增加导致细胞间基质被分解。茶叶作为广为人知的保健饮品,其在抗皮肤光老化方面的作用已得到了证实。茶叶水提物可以通过抑制MAPK磷酸化的途径来预防中波紫外线所致的皮肤光老化症状。文中还对该领域现有研究存在的不足进行了分析,旨在为下一步更全面深入的研究提供研究参考。     

Graded mode of transcriptional induction in yeast pheromone signalling revealed by single-cell analysis

Signalling pathways typically convert a graded, analogue signal into a binary cellular output. In the several eukaryotic systems that have been investigated to date, including MAP kinase cascade activation in Xenopus oocytes, analogue-to-digital conversion occurs at points in the pathway between receptor activation and the effector mechanism. We used flow cytometry combined with an intracellular fluorescent reporter to examine the characteristics of the yeast pheromone-response pathway. Surprisingly, pheromone response in yeast, which relies on the MAP kinase cascade, behaved in a fundamentally graded manner. Expression of certain exogenous dominant inhibitors of the pathway converted the response to graded-or-none behaviour. These results have implications for the dissection of biological response mechanisms in cells and illustrate how signalling pathways, even homologous ones, may have strikingly different signal propagation/amplification characteristics.     

KlROM2 encodes an essential GEF homologue in Kluyveromyces lactis

Cellular integrity in yeasts is ensured by a rigid cell wall whose synthesis is controlled by a MAP kinase signal transduction cascade. In Saccharomyces cerevisiae upstream regulatory components of this MAP kinase pathway involve a single protein kinase C, which is regulated in part by interaction with the small GTPase Rho1p. This small G protein is in turn rendered inactive (GDP-bound) or is activated (GTP-bound) by the influence of GTPase activating proteins (GAPs) and the GDP/GTP exchange factors (GEFs), respectively. We report here on the isolation of a gene from Kluyveromyces lactis, KlROM2, which encodes a member of the latter protein family. The nucleotide sequence contains an open reading frame of 1227 amino acids, with an overall identity of 57% to the Rom2 protein of S. cerevisiae. Four conserved sequence motifs could be identified: a RhoGEF domain, a DEP sequence, a CNH domain and a less conserved pleckstrin homology (PH) sequence. Klrom2 null mutants show a lethal phenotype, which indicates that the gene may encode the only functional GEF regulating the cellular integrity pathway in K. lactis. Conditional genomic expression of KlROM2 resulted in sensitivity towards caffeine and Calcofluor white as typical phenotypes of mutants defective in this pathway. Overexpression of KIROM2 from multicopy plasmids under the control of the ScGAL1 promoter severely impaired growth in both S. cerevisiae and in K. lactis. The fact that the lethal phenotype was not prevented in mpk1 deletion mutants indicates that growth inhibition is not simply caused by hyperactivation of the Pkc1p signal transduction pathway.     

Time-course regulation of quercetin on cell survival/proliferation pathways in human hepatoma cells

Quercetin, a dietary flavonoid, has been shown to possess anticarcinogenic properties, but the precise molecular mechanisms of action are not thoroughly elucidated. This study was aimed at investigating the time-course regulation effect of quercetin on survival/proliferation pathways in a human hepatoma cell line (HepG2). Quercetin induced a significant time-dependent inactivation of the major survival signaling proteins, i. e., phosphatidylinositol 3-kinase (PI 3-kinase)/protein kinase B (AKT), extracellular regulated kinase (ERK), protein kinase C-alpha (PKC-alpha), in concert with a time-dependent activation of key death-related signals: c-jun amino-terminal kinase (JNK) and PKC-delta. These data suggest that quercetin exerts a tight regulation of survival/proliferation pathways that requires the integration of different signals and persists over time, being the balance of these regulatory signals what determines the fate of HepG2 cells.     

Effects of a synthetic di-phosphoserine peptide (SS-2) on gene expression profiling against TNF-α induced inflammation

It has been showed bioactive di-phosphoserine peptide (SS-2) possesses functions to ameliorate oxidative stress in vitro. This study aimed to substantiate the role of bioactive di-phosphoserine peptide (SS-2) in modulating inflammatory responses in TNF-α-stimulated HT-29 cells, and its mechanism of action. SS-2 significantly reduced IL-8 secretion in TNF-α-induced HT-29 cells, and also suppressed pro-inflammatory cytokines, including IL-8, IL-12, MCP-1 and TNF-α. Moreover, SS-2 inhibited TNF-α initiated signalling cascades by suppressing phosphorylation of the ERK1/2, JNK, P38 and IκB those culminate in above cellular inflammatory responses. Differentially expressed genes analysis within NF-κB signalling pathway revealed that SS-2 blocks multiple sites of upstream NF-κB signalling cascade, including FADD and MyD88, thereby preventing the signalling transduction involved in cellular inflammatory response. These results provide a new insight into molecular mechanism for anti-inflammatory action of SS-2, suggesting SS-2 is a potential alternative approach to treat IBD by particular targeting TNF-α driven inflammatory event.     

Possible role of extracellular signal-regulated kinase pathway in regulation of Sox9 mRNA expression in chondrocytes under hydrostatic pressure

The potential involvement of the extracellular signal-regulated kinase (ERK) pathway in chondrocyte mechanotransduction was tested in bovine chondrocyte-agarose constructs under hydrostatic loading. Results suggested that the ERK pathway may be inhibited by hydrostatic pressure-induced mechanotransduction and may also be a negative regulator of Sox9 mRNA expression, which is an important modulator of chondrocyte function.     

宰后肌肉能量代谢对肉品质的影响研究进展

肉色、嫩度、保水性等肉的品质特性一直是影响消费者购买意愿的重要因素,而肌肉能量代谢会影响肌肉的生长、发育、成熟过程,并最终影响肉的食用品质。磷酸肌酸系统、糖酵解途径和氧化磷酸化是肌肉中ATP主要来源,并在宰后不同时期发挥主要作用。动物屠宰放血后,肌肉内环境发生变化,不同的供能方式会影响能量代谢过程,从而影响肌肉转化为“肉”的过程。本文综述宰后肌肉能量代谢过程及其对肉品质的影响,并阐述腺苷酸活化蛋白激酶（AMP-activated protein kinase,AMPK）和沉默信息调节因子（silent information regulators,SIRTs）家族对代谢过程的影响,以期为通过能量代谢过程调控肉品质的形成提供新的思路。     

人源乳杆菌联合菊粉对2型糖尿病小鼠的改善作用

目的：探究人源乳杆菌联合菊粉对2型糖尿病小鼠的改善效果及机制。方法：采用db/db糖尿病模型小鼠,灌胃4 株人源乳杆菌混合菌剂（10 mL/（kg mb·d）,每株菌的活菌数约1.0×109 CFU/mL）,基础饲料添加质量分数5%菊粉,对小鼠进行饮食干预,每周测定小鼠体质量、摄食量和空腹血糖浓度;干预末期进行小鼠口服葡萄糖耐量试验和胰岛素耐受试验;实验结束后测定小鼠空腹血糖浓度、血脂指标、糖化血清蛋白浓度、炎症因子mRNA相对表达水平、磷脂酰肌醇-3-激酶（phosphatidylinositol-3-kinase,PI3K）和丝氨酸/苏氨酸激酶（serine/threonine kinase,AKT）相对表达水平;取小鼠胰腺、肾脏和肝脏采用苏木精-伊红染色并观察组织形态。结果：人源乳杆菌联合菊粉干预可有效调节糖尿病模型小鼠血糖浓度和血脂指标水平,降低胰岛素抵抗和糖化血清蛋白浓度;抑制促炎因子白细胞介素（interleukin,IL）-6和IL-1β的mRNA表达,促进抑炎因子IL-10的mRNA表达;上调PI3K和AKT蛋白的表达;对肝脏、肾脏和胰腺的损伤有明显的改善作用。结论：人源乳杆菌联合菊粉通过有效降低胰岛素抵抗、抑制炎症反应和促进PI3K/AKT信号通路表达改善2型糖尿病小鼠症状。     

龙眼蛋白对C57BL/6小鼠及RAW264.7巨噬细胞的炎症因子的影响研究

目的 探讨龙眼蛋白对C57BL/6小鼠及RAW264.7巨噬细胞的炎症因子影响及作用机制。方法 采用反向色谱-质谱法(reverse-phase liquid chromatography-mass spectrometry,RPLC-MS)鉴定龙眼蛋白的组成。采用100 mg/(kg·d)和200 mg/(kg·d)龙眼蛋白腹腔注射C57BL/6小鼠,酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定小鼠血液炎症因子的表达,苏木精-伊红(hematoxylin-eosin,HE)染色法染色分析肺部炎症病理反应;用0.2、0.4、0.6 mg/mL龙眼蛋白处理小鼠RAW264.7巨噬细胞,噻唑蓝[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]法测定细胞活力,实时荧光定量聚合酶链式反应法(real-time quantitative polymerase chain reaction,Q-PCR)和ELISA检测炎症因子表达,蛋白质免疫印迹法(w...     

Phenolic Antioxidant Biosynthesis in Plants for Functional Food Application: Integration of Systems Biology and Biotechnological Approaches

We are applying a dynamic systems biology approach to the development of several phenolic phytochemicals in food-grade plants as ingredients for functional food applications. Phenolic antioxidant phytochemicals from food-grade plants will be an important part of a healthy diet in a global population that is projected to reach 9 billion in the next 50 years. Such phytochemicals are being targeted for designing conventional foods with added health benefits (functional foods). Such value-added foods are needed for dietary support to manage major oxidation-linked diseases, such as diabetes, cardiovascular disease, arthritis, cognition diseases and cancer. Plants produce phenolic metabolites as a part of growth, developmental and stress-adaptation response. These stress and developmental-modulated phenolic phytochemicals can be targeted for the design of functional foods. In order to design consistent food-grade phytochemical profiles for safety and clinical relevancy, novel tissue culture and bioprocessing technologies have been developed. The strategy for designing these phenolic phytochemicals is based on the model that phenolic metabolites in plants are efficiently produced through an alternative mode of metabolism that links proline synthesis to activity of the pentose-phosphate pathway. Using the proline-linked pentose-phosphate pathway model, techniques have been developed to isolate high phenolic clonal lines of food-grade plants from single heterozygous seeds. Further, using the same model, elicitation concepts and techniques have been applied to over-produce phenolic metabolites in seeds and sprouts. In both clonal and seed sprout systems, exogenous treatment of phenolic phytochemicals from a non-target species elicited endogenous stimulation of phenolic synthesis and, potentially, an antioxidant response. From these investigations, a hypothetical model has been proposed in which the proline-linked pentose-phosphate pathway is critical for modulating protective antioxidant response pathways in diverse biological systems, including humans. This model, when confirmed precisely, may provide a mechanism for understanding the mode of action of phenolic phytochemicals in modulating antioxidant pathways in relation to human health. An understanding of the interconnection of the proline-linked pentose-phosphate pathway and antioxidant response pathway can provide dietary and nutritional mechanisms as well as new strategies to manage oxidation-linked diseases through improvement of host physiological response. In this review we have focused on clonal herbs, fava bean sprouts and cranberry bioprocessing as 3 model systems for understanding biosynthesis of phenolic metabolites for functional food application.     

Effects of nitric oxide on the alleviation of postharvest disease induced by Penicillium italicum in navel orange fruits

The present study evaluated the effects of 15 μL L^-1 nitric oxide (NO) on the fruit defence response of ‘Newhall’ navel orange. The decay rate of NO-treated navel orange fruits was significantly lower than that of control fruits during storage at 20 °C storage (P < 0.05). Treatment with NO suppressed the increase in disease incidence and lesion area in orange fruits inoculated with Penicillium italicum (P. italicum); significantly increased the activities of phenolic metabolism-associated enzymes and pathogenesis-related (PR) proteins, including polyphenoloxidase (PPO), phenylalanine ammonia-lyase (PAL), β-1,3-glucanase (GLU) and chitinase (CHT); and enhanced the activities of key enzymes, including 4-coumarate: CoA ligase (4CL), cinnamate-4-hydroxylase (C4H) and chalcone isomerase (CHI) in the phenylpropanoid pathway. The contents of total phenolics, flavonoids and lignin were also higher in NO-treated fruits than in control fruits. The findings suggest that exogenous NO could induce disease resistance against P. italicum in navel orange fruits.