Impaired Thymic Export and Increased Apoptosis Account for Regulatory T Cell Defects in Patients with Non-ST Segment Elevation Acute Coronary Syndrome

Regulatory T (Treg) cells play a protective role against the development of atherosclerosis. Previous studies have revealed Treg cell defects in patients with non-ST elevation acute coronary syndrome (NSTACS), but the mechanisms underlying these defects remain unclear. In this study, we found that the numbers of peripheral blood CD4⁺CD25⁺CD127^low Treg cells and CD4⁺CD25⁺CD127^lowCD45RA⁺CD45RO⁻ naive Treg cells were lower in the NSTACS patients than in the chronic stable angina (CSA) and the chest pain syndrome (CPS) patients. However, the number of CD4⁺CD25⁺CD127^lowCD45RA⁻CD45RO⁺ memory Treg cells was comparable in all of the groups. The frequency of CD4⁺CD25⁺CD127^lowCD45RO⁻CD45RA⁺CD31⁺ recent thymic emigrant Treg cells and the T cell receptor excision circle content of purified Treg cells were lower in the NSTACS patients than in the CSA patients and the CPS controls. The spontaneous apoptosis of Treg cells (defined as CD4⁺CD25⁺CD127^lowannexin V⁺7-AAD⁻) was increased in the NSTACS patients compared with the CSA and CPS groups. Furthermore, oxidized LDL could induce Treg cell apoptosis, and the oxidized LDL levels were significantly higher in the NSTACS patients than in the CSA and CPS groups. In accordance with the altered Treg cell levels, the concentration of TNF-α was increased in the NSTACS patients, resulting in a decreased IL-10/TNF-α ratio. These findings indicate that the impaired thymic output of Treg cells and their enhanced susceptibility to apoptosis in the periphery were responsible for Treg cell defects observed in the NSTACS patients.     

RAP80 protein is important for genomic stability and is required for stabilizing BRCA1-A complex at DNA damage sites in vivo

RAP80 (receptor-associated protein 80) is a ubiquitin-binding protein that can specifically recognize and bind to Lys-63-linked polyubiquitin chains, thus targeting the BRCA1-A complex to DNA damage sites. To study the role of RAP80 in vivo, we generated RAP80-deficient mice. The deficient mice are prone to B-cell lymphomagenesis. B-cell lymphomas in RAP80-deficient mice are nearly diploid but harbor clonal chromosome translocations. Moreover, the deficient mice are hypersensitive to ionizing radiation. Repair of ionizing radiation-induced DNA double-strand breaks is impaired in RAP80-deficient mouse embryonic fibroblasts. Mechanistically, loss of RAP80 suppresses recruitment of the BRCA1-A complex to DNA damage sites and abrogates the DNA damage repair process at DNA damage sites. Taken together, these results reveal that RAP80 plays a crucial role in the DNA damage response and in maintaining genomic integrity.     

Novel triazines as potent and selective phosphodiesterase 10A inhibitors

The identification of highly potent and orally active triazines for the inhibition of PDE10A is reported. The new analogs exhibit low-nanomolar potency for PDE10A, demonstrate high selectivity against all other members of the PDE family, and show desired drug-like properties. Employing structure-based drug design approaches, we investigated the selectivity of PDE10A inhibitors against other known PDE isoforms, by methodically exploring the various sub-regions of the PDE10A ligand binding pocket. A systematic assessment of the ADME and pharmacokinetic properties of the newly synthesized compounds has led to the design of drug-like candidates with good brain permeability and desirable drug kinetics (t(1/2), bioavailability, clearance). Compound 66 was highly potent for PDE10A (IC(50)=1.4nM), demonstrated high selectivity (>200×) for the other PDEs, and was efficacious in animal models of psychoses; reversal of MK-801 induced hyperactivity (MED=0.1mg/kg) and conditioned avoidance responding (CAR; ID(50)=0.2mg/kg).     

利用定点突变法研究精氨酸脱亚胺酶活性的影响机制

精氨酸脱亚胺酶(ADI)是一种针对精氨酸缺陷型癌症(如:肝癌、黑素瘤)的新药,目前处于临床三期试验。文中通过定点突变技术分析了精氨酸脱亚胺酶的特定氨基酸位点对酶活力的影响机制。针对已报道的关键氨基酸残基A128、H404、I410,采用QuikChange法进行定点突变,获得ADI突变株M1(A128T)、M2(H404R)、M3(I410L)和M4(A128T/H404R)。将突变株在大肠杆菌BL21(DE3)中进行重组表达,并对纯化获得的突变蛋白进行酶学性质研究。结果表明,突变位点A128T和H404R对ADI最适pH的提高,生理中性(pH 7.4)条件下的酶活力和稳定性的提高,以及Km值的降低均具有显著的作用。研究结果为阐明ADI的酶活力影响机制和蛋白质的理性改造提供了一定的依据。     

模拟水分胁迫对不同种源麻楝种子萌发能力的影响

以麻楝6个种源种子为试验材料,用不同浓度的聚乙二醇(PEG)溶液模拟干旱胁迫,探讨干旱胁迫对种子发芽率、发芽势、发芽指数、活力指数以及幼苗苗高和胚根长及根苗比的影响,为麻楝的引种和推广种植提供依据。结果显示:(1)不同水势胁迫处理均降低了麻楝种子的发芽率和发芽势,当水势为-0.40MPa时延缓了种子萌发进程;种子的发芽率、发芽势、发芽指数和活力指数均随干旱胁迫强度的增加呈明显下降的趋势;当胁迫水势为-0.86MPa时,干旱胁迫处理的种子在试验结束时仍未能萌发,即-0.86MPa是麻楝种子萌发的临界水势。(2)当胁迫水势高于-0.40MPa时,麻楝幼苗的胚根长度与对照组差异不显著且长于对照组,说明高于-0.40MPa的水势有利于麻楝种子胚根的生长;麻楝幼苗苗高生长则是随着PEG浓度的升高而逐渐减缓。(3)适当的干旱胁迫可以增大各种源麻楝幼苗根苗比,且在胁迫水势高于-0.20MPa时都达到最大值。研究表明,麻楝种子具有一定的抗干旱胁迫的萌发能力,并以来自缅甸的Khin Aye Pale和泰国的Phu Wiang材料较强,来源于中国三亚和马来西亚Ulu Tranan的较弱。     

梵净山自然保护区珙桐天然种群生命表与生存分析

以种群生命表和生存分析理论为基础,采用空间代替时间法和分段匀滑技术,编制梵净山自然保护区珙桐天然种群特定时间生命表,绘制其死亡率曲线、消失率曲线、存活曲线和生存函数曲线,分析种群数量动态变化。结果表明:珙桐种群结构存在波动性,趋于DeeveyⅢ型,其幼年阶段的个体较丰富;珙桐种群死亡率和消失率曲线变化趋势基本一致,在这一发育过程中有两个死亡高峰,一个出现在幼苗向幼树的过渡期(Ⅰ龄级→Ⅱ龄级),另一个出现在从中龄向老龄过渡的阶段(Ⅷ龄级→Ⅸ龄级);种群生存分析表明,珙桐种群的生存率单调下降,累计死亡率单调上升,Ⅷ龄级后,种群生存率小于8%,累计死亡率大于92%,危险率超过生存率;4个生存函数曲线表明,梵净山珙桐有前期锐减、中期稳定和后期衰退的特点。幼苗和中龄级个体的不足是导致珙桐濒危的重要原因。     

Cloning and characterization of a novel amidase from <Emphasis Type="Italic">Paracoccus</Emphasis> sp. M-1, showing aryl acylamidase and acyl transferase activities

A novel amidase gene, designated pamh, was cloned from Paracoccus sp. M-1. Site-directed mutagenesis and bioinformatic analysis showed that the PamH protein belonged to the amidase signature enzyme family. PamH was expressed in Escherichia coli, purified, and characterized. The molecular mass of PamH was determined to be 52 kDa with an isoelectric point of 5.13. PamH displayed its highest enzymatic activity at 45°C and at pH 8.0 and was stable within a pH range of 5.0–10.0. The PamH enzyme exhibited amidase activity, aryl acylamidase activity, and acyl transferase activity, allowing it to function across a very broad substrate spectrum. PamH was highly active on aromatic and short-chain aliphatic amides (benzamide and propionamide), moderately active on amino acid amides, and possessed weak urease activity. Of the anilides examined, only propanil was a good substrate for PamH. For propanil, the k _cat and K _m were 2.8 s^?1 and 158 μM, respectively, and the catalytic efficiency value (k _cat/K _m) was 0.018 μM^?1 s^?1. In addition, PamH was able to catalyze the acyl transfer reaction to hydroxylamine for both amide and anilide substrates, including acetamide, propanil, and 4-nitroacetanilide; the highest reaction rate was shown with isobutyramide. These characteristics make PamH an excellent candidate for environmental remediation and an important enzyme for the biosynthesis of novel amides.     

植物根尖生长素信号研究进展

生长素信号调控植物生长发育的各个方面。该文综述了生长素信号在植物根尖的研究进展概况,从生长素在根尖的运输与分布、生长素信号对根尖细胞命运的影响及静止中心细胞的生长素信号研究三个方面进行了阐述,并对未来该领域的研究方向进行了展望。     

Stability and bioactivity studies on dipeptidyl peptidase IV resistant glucogan-like peptide-1 analogues

Glucagon-like peptide-1 (GLP-1) was once considered as an ideal anti-diabetic candidate for its important role in maintaining glucose homeostasis through the regulation of islet hormone secretion, as well as hepatic and gastric function. However, the major therapeutic obstacle for using native GLP-1 as a therapeutic agent is its very short half-life primarily due to their degradation by the enzyme dipeptidyl peptidase IV (DPP-IV). In this study, GLP-1 analogues with modifications in amino acid site 8, 22 and 23 were synthesized using solid phase peptide synthesis. Resistance of these analogues to DPP-IV cleavage was investigated in vitro by incubation of the peptides with DPP-IV or human plasma. Glucoregulating efficacy of the analogues was evaluated in normal Kunming mice using intraperitoneal glucose tolerance model. Glucose lowering effect of combination therapy (analogue plus Vildagliptin) has also been studied. In vitro studies showed that the modified analogues were much more stable than native GLP-1 (nearly 100% of the peptide keep intact after 4 h incubation). In vivo biological activity evaluation revealed that His8-EEE (the most potent GLP-1 analogues in this study) exhibited significantly improved glycemic control potency (approximately 4.1-fold over saline and 2.5-fold over GLP-1) and longer time of active duration (at least 5 h). Combination therapy also showed the trend of its superiority over mono-therapy. Modified analogues showed increased potency and biological half-time compared with the native GLP-1, which may help to understand the structure-activity relationship of GLP-1 analogues.     

A novel anticancer and antifungus phenazine derivative from a marine actinomycete BM-17

A marine actinomycete, designated strain BM-17, was isolated from a sediment sample collected in the Arctic Ocean. The strain was identified as Nocardia dassonvillei based on morphological, cultural, physiological, biochemical characteristics, along with the cell wall analysis and 16S rDNA gene sequence analysis. A new secondary metabolite (1), N-(2-hydroxyphenyl)-2-phenazinamine (NHP), and six known antibiotics (2–7) have been isolated from the saline culture broth of the stain by sequentially purification over macroporous resin D101, silica gel, Sephadex LH-20 column chromatography and preparative HPLC after the stain was incubated in soy bean media at 28 °C for 7 days. The chemical structures of the compounds were elucidated on the basis of spectroscopic analysis, including two-dimensional (2D) NMR and HR-ESI-MS data. The new compound showed significant antifungal activity against Candida albicans, with a MIC of 64 μg/ml and high cancer cell cytotoxicity against HepG2, A549, HCT-116 and COC1 cells.