Enzymatic activity and substrate specificity of mitogen-activated protein kinase p38alpha in different phosphorylation states

The mitogen-activated protein (MAP) kinases are essential signaling molecules that mediate many cellular effects of growth factors, cytokines, and stress stimuli. Full activation of the MAP kinases requires dual phosphorylation of the Thr and Tyr residues in the TXY motif of the activation loop by MAP kinase kinases. Down-regulation of MAP kinase activity can be initiated by multiple serine/threonine phosphatases, tyrosine-specific phosphatases, and dual specificity phosphatases (MAP kinase phosphatases). This would inevitably lead to the formation of monophosphorylated MAP kinases. However, the biological functions of these monophosphorylated MAP kinases are currently not clear. In this study, we have prepared MAP kinase p38alpha, a member of the MAP kinase family, in all phosphorylated forms and characterized their biochemical properties. Our results indicated the following: (i) p38alpha phosphorylated at both Thr-180 and Tyr-182 was 10-20-fold more active than p38alpha phosphorylated at Thr-180 only, whereas p38alpha phosphorylated at Tyr-182 alone was inactive; (ii) the dual-specific MKP5, the tyrosine-specific hematopoietic protein-tyrosine phosphatase, and the serine/threonine-specific PP2Calpha are all highly specific for the dephosphorylation of p38alpha, and the dephosphorylation rates were significantly affected by different phosphorylated states of p38alpha; (iii) the N-terminal domain of MPK5 has no effect on enzyme catalysis, whereas deletion of the MAP kinase-binding domain in MKP5 leads to a 370-fold decrease in k(cat)/K(m) for the dephosphorylation of p38alpha. This study has thus revealed the quantitative contributions of phosphorylation of Thr, Tyr, or both to the activation of p38alpha and to the substrate specificity for various phosphatases.     

A method for simultaneous gene overexpression and inactivation in the <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis> genome

The gene integration method is an important tool to stably express desirable genes in bacteria. To avoid heavy workload and cost, we constructed a rapid and efficient method for genome modification. This method depended on a mobilizable plasmid, which contains a P _tac promoter, an introduced multiple cloning site (iMCS), and rrnBT1T2 terminator. Briefly, the mobilizable plasmid pK18-MBPMT with the P _tac-iMCS-rrnBT1T2 cartridge derived from pK18mobsacB was prepared to directly integrate hetero-/homologous DNA into the Corynebacterium glutamicum genome. Like our previous method, this method was based on insertional inactivation and double-crossover homologous recombination, which simultaneously achieved gene overexpression and inactivation in the genome without the use of genetic markers. Compared to the previous method, this protocol omitted the construction of a recombinant expression plasmid and clone of the target gene(s) cassette, which significantly decreased the workload, cost, and operational time. Using this method, the heterologous gene amy and the homologous gene lysC ^T311I were successfully integrated into the C. glutamicum genome at alaT and avtA loci, respectively. Moreover, the operation time of this method was shorter than that of the previous method, especially for repeated integration. This method, which is based on the mobilizable plasmid pK18-MBPMT, thus represents a potentially attractive protocol for the integration of genes in the course of genetic modification of C. glutamicum.     

中国鬼伞类真菌的分类

鬼伞类真菌涉及蘑菇科的鬼伞属Coprinus以及小脆柄菇科的小鬼伞属Coprinellus、拟鬼伞属Coprinopsis、近地伞属Parasola、刺毛鬼伞属Tulosesus和心孢鬼伞属Narcissea。基于标本研究和文献记载,确认我国该类真菌57种：小鬼伞属9种,刺毛鬼伞属10种,心孢鬼伞属3种,拟鬼伞属25种,近地伞属8种,鬼伞属2种。本文记载1个新组合——速亡型心孢鬼伞Narcissea ephemeroides,中国新记录种11个：黄鳞小鬼伞Coprinellus ellisii、甜味小鬼伞Coprinellus saccharinus、锐突拟鬼伞Coprinopsis acuminata、非洲雪白拟鬼伞Coprinopsis afronivea、美丽拟鬼伞Coprinopsis bellula、钟孢拟鬼伞Coprinopsis mitraespora、麻醉拟鬼伞Coprinopsis narcotica、厚壁拟鬼伞Coprinopsis pachyderma、近雪白拟鬼伞Coprinopsis pseudonivea、施罗特近地伞Parasola schroeteri和刺毛近地伞Parasola setulosa。编制了中国鬼伞类真菌分种检索表,对新组合和中国新记录种进行形态学描述,并提供线条图。结合ITS和LSU序列片段,采用了最大似然法和贝叶斯分析法建立系统发育树,反映各类群之间的系统演化关系。     

An integrated fluid–structure interaction and thrombosis model for type B aortic dissection

False lumen thrombosis (FLT) in type B aortic dissection has been associated with the progression of dissection and treatment outcome. Existing computational models mostly assume rigid wall behavior which ignores the effect of flap motion on flow and thrombus formation within the FL. In this study, we have combined a fully coupled fluid–structure interaction (FSI) approach with a shear-driven thrombosis model described by a series of convection–diffusion reaction equations. The integrated FSI-thrombosis model has been applied to an idealized dissection geometry to investigate the interaction between vessel wall motion and growing thrombus. Our simulation results show that wall compliance and flap motion can influence the progression of FLT. The main difference between the rigid and FSI models is the continuous development of vortices near the tears caused by drastic flap motion up to 4.45 mm. Flap-induced high shear stress and shear rates around tears help to transport activated platelets further to the neighboring region, thus speeding up thrombus formation during the accelerated phase in the FSI models. Reducing flap mobility by increasing the Young’s modulus of the flap slows down the thrombus growth. Compared to the rigid model, the predicted thrombus volume is 25% larger using the FSI-thrombosis model with a relatively mobile flap. Furthermore, our FSI-thrombosis model can capture the gradual effect of thrombus growth on the flow field, leading to flow obstruction in the FL, increased blood viscosity and reduced flap motion. This model is a step closer toward simulating realistic thrombus growth in aortic dissection, by taking into account the effect of intimal flap and vessel wall motion.

     

Reprogramming of sugar transport pathways in Escherichia coli using a permeabilized SecY protein-translocation channel

In the initial step of sugar metabolism, sugar-specific transporters play a decisive role in the passage of sugars through plasma membranes into cytoplasm. The SecY complex (SecYEG) in bacteria forms a membrane channel responsible for protein translocation. The present work shows that permeabilized SecY channels can be used as nonspecific sugar transporters in Escherichia coli. SecY with the plug domain deleted allowed the passage of glucose, fructose, mannose, xylose, and arabinose, and, with additional pore-ring mutations, facilitated lactose transport, indicating that sugar passage via permeabilized SecY was independent of sugar stereospecificity. The engineered E. coli showed rapid growth on a wide spectrum of monosaccharides and benefited from the elimination of transport saturation, improvement in sugar tolerance, reduction in competitive inhibition, and prevention of carbon catabolite repression, which are usually encountered with native sugar uptake systems. The SecY channel is widespread in prokaryotes, so other bacteria may also be engineered to utilize this system for sugar uptake. The SecY channel thus provides a unique sugar passageway for future development of robust cell factories for biotechnological applications.     

贵州省野生苦苣苔科物种多样性与地理分布（附录）

该研究在建立贵州省野生苦苣苔科植物名录和地理分布数据库的基础上,对其物种多样性及地理分布格局进行研究。通过文献资料结合实地调查,从物种组成、特有性、水平分布、垂直分布和相似性等方面进行分析,并采用筛除算法确定贵州苦苣苔科植物分布的热点地区。结果表明:(1)贵州省苦苣苔科植物共计2族8亚族28属153种(含种下等级),分布在75个县级行政区,有128/45个中国/贵州特有种,垂直分布以900~1300 m海拔段最为丰富。(2)通过计算省级相似性系数,发现贵州与广西的相似程度最高,最后筛选得到10个热点县,共代表了75%的苦苣苔科植物。(3)贵州省为典型的喀斯特高原山地,苦苣苔科植物种类丰富,尤其是广义马铃苣苔属、广义报春苣苔属、广义石山苣苔属和蛛毛苣苔属等,有着较高的物种多样性和区域特有性。该研究可以为贵州省苦苣苔科植物资源保护和持续利用提供理论参考。     

XIAP及survivin 在原发性肝细胞癌组织中的表达及其相关性研究

目的：探讨X 连锁凋亡抑制蛋白（XIAP）和survivin 在原发性肝细胞癌中的表达及两者的相关性。方法：选取本院收治的60 例原发性肝细胞癌患者,应用免疫组织化学染色的方法对肝癌组织及癌旁组织中的XIAP及survivin 的表达进行检测。结果：经 比较,肝癌组织中XAIP 及survivin 的阳性率均显著高于癌旁组织,差异有统计学意义。XIAP 和survivin 的表达强度与肿瘤的大 小无关,但随肿瘤的分化程度的降低而升高,且不同分化程度之间差异有统计学意义;XIAP 和survivin 存在正相关关系。结论： XIAP与Survivin 在肿瘤组织中的高表达在促进肿瘤发生、增殖、转移以及耐药,并且能够降低肿瘤的分化程度,增加肝癌的恶性 程度。此外,两者可能存在协同作用,但两者的相关性及作用机制仍需进一步探讨。     

不同密度格木幼林的土壤理化与林下植被特征

为探索适合格木(Erythrophleum fordii)人工林在幼龄阶段的种植密度,在不同林分密度(2 m×1 m、2 m×2 m、2 m×3 m、3 m×3 m)的6 a生格木人工林下设置标准样地,采用土壤质量评价和灰色关联度等方法,探究不同密度下格木幼林的土壤理化与林下植被特征。结果表明,密度2 m×3 m下的林木胸径、树高最优,较最低水平高16.7%、27.9%;土壤总孔隙度最大,全N、硝态N、铵态N含量最高,灌木草本多样性最高。相关性分析表明土壤化学性质对灌木草本的多样性影响最大。不同林分密度下格木幼林土壤理化性质及林下植物多样性有显著差异,因此,选择合适的林分密度对人工林土壤肥力的可持续利用及林分的经营培育至关重要。     

Role of bone marrow-derived mesenchymal stem cells in the prevention of hyperoxia-induced lung injury in newborn mice

BPD (bronchopulmonary dysplasia) is predominantly characterized by persistent abnormalities in lung structure and arrested lung development, but therapy can be palliative. While promising, the use of BMSC (bone marrow-derived mesenchymal stem cell) in the treatment of lung diseases remains controversial. We have assessed the therapeutic effects of BMSC in vitro and in vivo. In vitro co-culturing with injured lung tissue increased the migration-potential of BMSC; and SP-C (surfactant protein-C), a specific marker of AEC2 (type II alveolar epithelial cells), was expressed. Following intraperitoneal injection of BMSC into experimental BPD mice on post-natal day 7, it was found that BMSC can home to the injured lung, express SP-C, improve pulmonary architecture, attenuate pulmonary fibrosis and increase the survival rate of BPD mice. This work supports the notion that BMSC are of therapeutic benefit through the production of soluble factors at bioactive levels that regulate the pathogenesis of inflammation and fibrosis following hyperoxia.