G蛋白偶联受体与G蛋白相互作用的最新研究进展

传统观念认为,在激动剂作用下,G蛋白偶联受体(GPCRs)能够激活G蛋白的α亚基,从而使Gα亚基与Gβγ亚基分离,被激活的Gα亚基通过信号转导进一步参与细胞的生理过程。但是,最新研究发现GPCRs和G蛋白存在多种偶联关系,GPCRs不仅能够激活Gα亚基,还可以与Gβγ亚基相互靠近,甚至会使G蛋白亚基构象发生重排而不分离,这对于疾病发病机制的研究及新的药物靶点的发现具有重要意义。就GPCRs与G蛋白之间的相互作用以及最新研究技术作一简要综述。     

北方农牧交错带温性盐碱化草地土壤呼吸对不同形态氮添加和刈割的响应

农牧交错带草地生态系统兼受农业和牧业的影响, 属于脆弱生态系统, 尤其是养分贫瘠的盐碱化草地, 其生态系统结构和功能对外界干扰的响应更加强烈。位于晋西北地区的农牧交错带盐碱化草地, 地理位置独特, 区别于天然牧区草地生态系统。由于毗邻农田, 农业氮肥的过量使用促进了活性氮气体排放, 同时使得农牧交错带草地土壤碳氮循环发生改变。刈割是北方农牧交错草地生态系统的主要管理方式, 为了深入探究氮添加和刈割管理方式对农牧交错带草地碳循环的影响, 进一步厘清该区域草地生态系统的碳动态问题, 该研究设置了一个不同形态氮添加和刈割的裂区实验, 测定土壤呼吸对不同形态氮肥添加和刈割的响应, 为进一步科学管理该区域草地提供可靠的依据。实验样地位于山西省右玉县境内的“山西农业大学农牧交错带草地生态系统野外观测研究站”, 于2017年设置不同形态氮添加和刈割处理, 实验处理包括对照(不刈割和刈割)、尿素添加、缓释尿素添加、刈割+尿素添加、刈割+缓释尿素添加, 每种处理6个重复, 共36个小区。在不同处理条件下测定土壤呼吸速率、土壤温度、土壤水分、土壤微生物生物量、土壤无机氮含量、植物地上和地下生物量, 并计算土壤累积碳排放量及CO₂通量。研究结果表明: (1)短期(2017-2018年)尿素和缓释尿素的添加显著提高了该地区土壤呼吸速率和土壤累积碳排放量。与添加缓释尿素相比, 添加尿素处理下的土壤呼吸速率和累积碳排放量更高; (2)刈割显著降低土壤呼吸速率和累积碳排放量; (3)短期氮添加和刈割的交互作用对土壤呼吸速率没有显著影响。因此, 短期氮添加促进了北方农牧交错带盐碱化草地土壤碳释放, 刈割抑制土壤呼吸, 降低了累积碳排放量, 这可能是由于刈割移除地上植物, 减少了凋落物的输入, 底物减少导致土壤微生物活性降低。但是随着处理时间的延长, 氮添加和刈割对该农牧交错带盐碱化草地土壤碳动态的影响还有待进一步探究和发现。     

上海公园水体夏季浮游植物群落与环境因子的关系

为了解公园水体浮游植物群落状况,分析浮游植物物种分布和环境因子之间的关系,揭示浮游植物物种对生态环境的需求,于2008年7月和9月对上海市11个公园水体浮游植物群落进行了调查,对获得的浮游植物数据和环境因子数据进行典范相关分析(CCA),并绘制了物种与环境因子关系的二维排序图。结果表明：调查期间共鉴定出浮游植物384种,隶属于8门,浮游植物密度范围为2.01×10⁵～57.60×10⁵ cells·L^-1;群落组成以蓝藻、裸藻、硅藻和绿藻为主,主要优势种有细微颤藻、无常蓝纤维藻、尾裸藻、颗粒直链藻、梅尼小环藻、普通小球藻、四尾栅藻等;7月影响浮游植物分布的主要环境因子依次为铵态氮、溶解氧、水温和总磷,而9月的pH值、水温、溶解氧、透明度和总氮含量对浮游植物的分布产生影响较大;其中,透明度和浮游动物量是影响隐藻、甲藻和硅藻藻类生物量的主要环境因子,而蓝藻、裸藻、绿藻主要受水体氮磷营养盐浓度和溶解氧的影响。     

Endogenous 3,4-Dihydroxyphenylalanine and Dopaquinone Modifications on Protein Tyrosine: LINKS TO MITOCHONDRIALLY DERIVED OXIDATIVE STRESS VIA HYDROXYL RADICAL*

Oxidative modifications of protein tyrosines have been implicated in multiple human diseases. Among these modifications, elevations in levels of 3,4-dihydroxyphenylalanine (DOPA), a major product of hydroxyl radical addition to tyrosine, has been observed in a number of pathologies. Here we report the first proteome survey of endogenous site-specific modifications, i.e. DOPA and its further oxidation product dopaquinone in mouse brain and heart tissues. Results from LC-MS/MS analyses included 50 and 14 DOPA-modified tyrosine sites identified from brain and heart, respectively, whereas only a few nitrotyrosine-containing peptides, a more commonly studied marker of oxidative stress, were detectable, suggesting the much higher abundance for DOPA modification as compared with tyrosine nitration. Moreover, 20 and 12 dopaquinone-modified peptides were observed from brain and heart, respectively; nearly one-fourth of these peptides were also observed with DOPA modification on the same sites. For both tissues, these modifications are preferentially found in mitochondrial proteins with metal binding properties, consistent with metal-catalyzed hydroxyl radical formation from mitochondrial superoxide and hydrogen peroxide. These modifications also link to a number of mitochondrially associated and other signaling pathways. Furthermore, many of the modification sites were common sites of previously reported tyrosine phosphorylation, suggesting potential disruption of signaling pathways. Collectively, the results suggest that these modifications are linked with mitochondrially derived oxidative stress and may serve as sensitive markers for disease pathologies.Generation of reactive oxygen species (ROS)¹ and reactive nitrogen species is a normal consequence of aerobic metabolism that, in excess, results in oxidative stress that further leads to oxidative modification of proteins, lipids, and DNA, events that may lead to altered cellular function and even cell death (1, 2). Chronic oxidative stress is well recognized as having a central role in disease and is responsible for both direct alteration of biomolecular structure-function and compensatory changes in cellular processes (1–4). It is increasingly recognized that oxidative modifications of proteins can serve as potential biomarkers indicative of the physiological states and changes that occur during disease progression. Thus, the ability to quantitatively measure specific protein oxidation products has the potential to provide the means to monitor the physiological state of a tissue or organism, in particular any progression toward pathology. Given Parkinson disease (PD) as an example, a number of oxidative modifications on proteins pertinent to PD have been identified, further supporting the potential importance of oxidative modifications to disease pathogenesis (5).Many oxidative modifications on specific amino acid residues, such as protein carbonylation (6), cysteine S-nitrosylation (7–9), cysteine oxidation to sulfinic or sulfonic acid (10–12), methionine oxidation (13, 14), and tyrosine nitration (15–21) within complex protein mixtures, have been detected by MS-based proteomics; however, their low abundance levels within complex proteomes often hinder confident identification of these potentially significant modifications (22). For example, tyrosine nitration is a well studied post-translational modification mediated by peroxynitrite (ONOO⁻) or nitrogen dioxide (^·NO₂), which commonly occur in cells during oxidative stress and inflammation; however, only a small number of nitrotyrosine proteins have been identified from a given proteome sample because of insufficient analytical sensitivity and the chance of incorrect peptide assignments (19, 23). With recent advances in high resolution MS that provide high mass measurement accuracy, the ability to confidently identify modified peptides has been significantly enhanced (24).Hydroxyl radical (HO^·) is one of the most reactive and major species generated under aerobic conditions in biological systems (1, 25, 26). Among several HO^·-mediated oxidative modifications, the protein tyrosine modification 3,4-dihydroxyphenylalanine (DOPA) has been reported as a major product and index of HO^· attack on tyrosine residues in proteins (Fig. 1) (27, 28). DOPA is also formed on protein tyrosine residues via controlled enzymatic pathways through enzymes such as tyrosinase or tyrosine hydroxylase (28). Once formed, protein-bound DOPA has the potential to initiate further oxidative reactions through binding and reducing transition metals or through redox cycling between catechol and quinone (dopaquinone) forms (29, 30). Recent studies have suggested that protein-bound DOPA is involved in triggering antioxidant defenses (30) and mediating oxidative damage to DNA (31). Moreover, elevated levels of protein-bound DOPA have been reported in several diseases, including atherosclerosis, cataracts, and myocardial disease, and in PD patients undergoing levodopa therapy (26, 32–36). However, the specific DOPA-modified proteins, which could provide mechanistic knowledge of the progression of these diseases, have not been identified (27, 28). The ability to identify site-specific protein modifications should lead to a better understanding of the role of DOPA modification in disease pathologies as well as new molecular signatures or therapeutic targets for diseases.Open in a separate windowFig. 1.DOPA and dopaquinone formation from tyrosine.Therefore, in this study, we demonstrate the ability to identify site-specific DOPA and dopaquinone (DQ) modifications on protein tyrosine residues in normal mouse brain and heart tissues and their relative stoichiometries that are present in vivo under non-stressed conditions. Such endogenous protein modifications were detected using LC-MS/MS. The results from this global proteomics survey suggests that HO^· in tissues under normal conditions is generated largely from the mitochondria and metal-binding proteins where the resulting DOPA/DQ modifications have the potential to disrupt mitochondrial respiration as well as alter tyrosine phosphorylation signaling pathways such as 14-3-3-mediated signaling in brain tissue.     

医院感染多重耐药革兰阴性杆菌耐消毒剂基因的检测分析

目的研究医院感染多重耐药革兰阴性杆菌耐消毒剂基因qac E△1-sul1存在情况。方法采用聚合酶链反应(PCR)技术检测qac E△1-sul1基因。结果 201株多重耐药革兰阴性杆菌对阿莫西林和头孢类等抗菌药物多数耐药率均达到50%以上,但对碳青霉烯类仍高度敏感。qac E△1-sul1基因的总检出率为40.80%,其中产超广谱β-内酰胺酶(ESBLs)大肠埃希菌、产ESBLs肺炎克雷伯菌、多重耐药鲍曼不动杆菌、多重耐药的铜绿假单胞菌qac E△1-sul1检出率分别为34.78%、44.23%、58.91%和31.25%。结论医院感染患者临床分离多重耐药革兰阴性杆菌耐消毒剂基因qac E△1-sul1携带率较高,加强多重耐药革兰阴性杆菌对消毒剂耐药性的监测,对临床合理使用消毒剂具有重要意义。     

The simultaneous determination of 1-aminocyclopropane-1-carboxylic acid and cyclopropane-1,1-dicarboxylic acid in Lycopersicum esculentum by high-performance liquid chromatography--electrospray tandem mass spectrometry

Varying concentrations of cyclopropane-1,1-dicarboxylic acid (CDA), an inhibitor of 1-aminocyclopropane-1-carboxylic acid oxidase, added to the solid culture medium of tomato nodal shoot segments resulted in a reduction in the level of endogenous ethylene according to the concentration of inhibitor applied. Following treatment with inhibitor, plants were homogenised and the concentrations of CDA and of 1-aminocyclopropane-1-carboxylic acid (ACC) were measured simultaneously in the resulting juice using an HPLC-ESI/MS-MS method. The levels of CDA and ACC measured in the plant tissues were associated with the concentration of inhibitor added to the solid medium. The HPLC-ESI/MS-MS method described produced limits of detection of 0.8 pmol for ACC and of 4 pmol for CDA.     

Bone marrow–derived mesenchymal stromal cells ameliorate severe acute pancreatitis in rats via hemeoxygenase-1–mediated anti-oxidant and anti-inflammatory effects

Background and aims

It has been previously verified that mesenchymal stromal cells (MSCs) have a good therapeutic effect on severe acute pancreatitis (SAP) and the potential for regeneration of damaged pancreatic tissue, but the exact molecular mechanism remains unclear. In this study, we demonstrated the therapeutic effect of bone morrow MSCs (BMSCs) on SAP, probably by targeting heme oxygenase-1 (HO-1).

Insulin-like growth factor 2 regulates the proliferation and differentiation of rat adipose-derived stromal cells via IGF-1R and IR

BackgroundInsulin-like growth factor 2 (IGF2), an essential component of the stem cell niche, has been reported to modulate the proliferation and differentiation of stem cells. Previously, a continuous expression of IGF2 in tissues was reported to maintain the self-renewal ability of several types of stem cells. Therefore, in this study, we investigated the expression of IGF2 in adipose tissues and explored the effects of IGF2 on adipose-derived stromal cells (ADSCs) in vitro.MethodsThe expression pattern of IGF2 in rat adipose tissues was determined by gene expression and protein analyses. The effect of IGF2 on proliferation, stemness-related marker expression and adipogenic and osteogenic differentiation was systematically investigated. Furthermore, antagonists of IGF2-specific receptors—namely, BMS-754807 and picropodophyllin—were added to explore the underlying signal transduction mechanisms.ResultsIGF2 levels displayed a tendency to decrease with age in rat adipose tissues. After the addition of IGF2, isolated ADSCs displayed higher proliferation and expression of the stemness-related markers NANOG, OCT4 and SOX2 and greater differentiation potential to adipocytes and osteoblasts. Additionally, both type 1 insulin-like growth factor receptor (IGF-1R) and insulin receptor (IR) participated in the IGF2-mediated promotion of stemness in ADSCs.ConclusionsOur findings indicate that IGF2 could enhance the stemness of rat ADSCs via IGF-1R and IR and may highlight an effective method for the expansion of ADSCs for clinical application.     

黑蛋巢菌属新种和新变种

本文报道了黑蛋巢菌属Cyathus四个新种和二个新变种,它们分别是:内蒙黑蛋巢菌Cyathus neimonggolensis Liu et Y.M.Li,盘柄黑蛋巢菌C.discostipltatus Liu et Y.M.Li,太原黑蛋巢菌C.taiyuanensis Liu et Y.M.Li,毛被黑蛋巢菌C.hirtulus Liu et Y.M.Li,天山黑蛋巢毛被变种C.tianshanensis Liu et Cao var.tomentosus Liu, Cao et Y.M.Li,环状黑蛋巢武夷山变种 C. annulatus Brodie var. wuyishanensis Liu et Y.M.Li。全部模式标本均保存在山西大学真菌标本室。