Interplay between hnRNP A1 and a cis-acting element in the 3' UTR of CYP2A5 mRNA is central for high expression of the gene

Our previous evidence suggests that heterogeneous nuclear ribonucleoprotein (hnRNP) A1 plays a part in the regulation of the Cyp2a5 gene by interacting with the 3' untranslated region (UTR) of the CYP2A5 mRNA. However, the exact role of this interaction is not clear. The aim of the present work was to gain further insight into the regulation process of Cyp2a5. For this purpose the 3' UTR of CYP2A5 was fused to the coding region of luciferase mRNA. Luciferase recombinants containing either the full length 3' UTR, or the 3' UTR lacking a previously described 71 nucleotide (nt) region (the hnRNP A1 primary binding site), were transiently expressed in cells expressing or lacking hnRNP A1. The expression of the luciferase recombinants was examined both at mRNA and enzyme activity levels. The results disclosed that the presence of hnRNP A1 was required for the high expression of the recombinant carrying the full length 3' UTR of CYP2A5. Deletion of the hnRNP A1 primary binding site dramatically modified the expression pattern: the mRNA levels and luciferase activities of the deletion mutant were independent from hnRNP A1. These results conclusively demonstrate that the 71 nt region in the 3' UTR of CYP2A5 mRNA can confer hnRNP A1-dependent regulation to a gene. In addition, comparison of RNA levels and luciferase activities suggested that regions flanking the hnRNP A1 binding site could regulate translation of the CYP2A5 mRNA. These results are consistent with a model in which the binding of hnRNP A1 to the 71 nt putative hairpin-loop region in the CYP2A5 mRNA 3' UTR upregulates mRNA levels possibly by protecting the mRNA from degradation.     

Lipopolysaccharide and proinflammatory cytokines stimulate interleukin-6 expression in C2C12 myoblasts: role of the Jun NH2-terminal kinase

IL-6 is a major inflammatory cytokine that plays a central role in coordinating the acute-phase response to trauma, injury, and infection in vivo. Although IL-6 is synthesized predominantly by macrophages and lymphocytes, skeletal muscle is a newly recognized source of this cytokine. IL-6 from muscle spills into the circulation, and blood-borne IL-6 can be elevated >100-fold due to exercise and injury. The purpose of the present study was to determine whether inflammatory stimuli, such as LPS, TNF-alpha, and IL-1beta, could increase IL-6 expression in skeletal muscle and C2C12 myoblasts. Second, we investigated the role of mitogen-activated protein (MAP) kinases, and the Jun NH2-terminal kinase (JNK) in particular, as a mediator of this response. Intraperitoneal injection of LPS in mice increased the circulating concentration of IL-6 from undetectable levels to 4 ng/ml. LPS also increased IL-6 mRNA 100-fold in mouse fast-twitch skeletal muscle. Addition of LPS, IL-1beta, or TNF-alpha directly to C2C12 myoblasts increased IL-6 protein (6- to 8-fold) and IL-6 mRNA (5- to 10-fold). The response to all three stimuli was completely blocked by the JNK inhibitor SP-600125 but not as effectively by other MAP kinase inhibitors. SP-600125 blocked LPS-stimulated IL-6 synthesis dose dependently at both the RNA and protein level. SP-600125 was as effective as the synthetic glucocorticoid dexamethasone at inhibiting IL-6 expression. SP-600125 inhibited IL-6 synthesis when added to cells up to 60 min after LPS stimulation, but its inhibitory effect waned with time. LPS stimulated IL-6 mRNA in both myoblasts and myotubes, but myoblasts showed a proportionally greater LPS-induced increase in IL-6 protein expression compared with myotubes. SP-600125 and the proteasomal inhibitor MG-132 blocked LPS-induced degradation of IkappaB-alpha/epsilon and LPS-stimulated expression of IkappaB-alpha mRNA. Yet, only SP-600125 and not MG-132 blocked LPS-induced IL-6 mRNA expression. This suggests that IL-6 gene expression is a downstream target of JNK in C2C12 myoblasts.     

Enhancement of intracellular calcium concentration by extracellular ATP and UTP in Madin Darby Canine Kidney cells

Fura2 - fluorescence was utilized to test for the effect of extracellular nucleotides on intracellular calcium concentration of subconfluent Madin-Darby Canine Kidney (MDCK)-cells. Extracellular ATP (10 mumol/l) and UTP (10 mumol/l) lead to rapid (within seconds), sustained, and fully reversible enhancement of intracellular calcium concentration from 138 +/- 9 nmol/l (n = 27), to 1561 +/- 260 nmol/l (n = 10) and 3435 +/- 949 nmol/l (n = 5), respectively. Half maximal effects are observed at some 1 mumol/l. In the absence of extracellular calcium the effect of ATP is transient, pointing to release of intracellular calcium. The sustained effect in the presence of extracellular calcium indicates that the nucleotides in addition recruit calcium from extracellular space.     

Types and Succession of Forest Bogs in Hingganling and Changbaishan

Climate and geomorphy arc the main factors of the formation of forest bogs in Hingganling and changbaishan. The bogs are both multiple in types and widespread in distribution. There exist not only the type of lower bog, but also that of middle bog and raised bog. They are chiefly distributed in the alluvial flat or valley and in the watershed. The succession process of forest bog is generalized into two kinds: (1) The formation of forest bogs as a result of the natural succession of forest vegetation under the influence of natural conditions. (2) The formation of forest bogs as a result of the bogginess in the vicinity of forestland.     

Central Coherence in Eating Disorders: A Synthesis of Studies Using the Rey Osterrieth Complex Figure Test

BackgroundLarge variability in tests and differences in scoring systems used to study central coherence in eating disorders may lead to different interpretations, inconsistent findings and between study discrepancies. This study aimed to address inconsistencies by collating data from several studies from the same research group that used the Rey Osterrieth Complex Figure Test (Rey Figure) in order to produce norms to provide benchmark data for future studies.MethodData was collated from 984 participants in total. Anorexia Nervosa, Bulimia Nervosa, recovered Anorexia Nervosa, unaffected family members and healthy controls were compared using the Rey Figure.ResultsPoor global processing was observed across all current eating disorder sub-groups and in unaffected relatives. There was no difference in performance between recovered AN and HC groups.ConclusionsThis is the largest dataset reported in the literature and supports previous studies implicating poor global processing across eating disorders using the Rey Figure. It provides robust normative data useful for future studies.     

不同食细菌线虫取食密度下线虫对细菌数量、活性及土壤氮素矿化的影响

采用悉生培养微缩体系,探讨了不同食细菌线虫取食密度下线虫(Caenorhabditis elegans) 对细菌(Bacillus subtilis)数量和活性及土壤氮素矿化的影响.结果表明,线虫对细菌的取食,促进了细菌的增殖,并在不同线虫取食密度下对细菌的增殖促进作用总体表现为:接种20条·g^-1＞10条·g^-1＞40条线虫·g^-1处理.线虫在促进细菌增殖的同时,明显提高了土壤呼吸强度和土壤蔗糖酶、脲酶和磷酸酶的活性,但不同取食密度处理间差异不明显.线虫与细菌之间的相互作用显著提高了土壤铵态氮和矿质态氮含量,促进了土壤氮的矿化.不同取食密度处理间,线虫对土壤氮素矿化的促进作用与对细菌的增殖促进作用趋势一致.     

艳山姜果实的生药学及气相色谱研究

艳山姜为姜科山姜属植物,艳山姜(Alpinia zerumbet)的干燥成熟果实,是贵州少数民族地区的传统习用药物,艳山姜在贵州省的种植面积已超过130 hm~2,是贵州"南药"的第一大宗产品,也是治理石漠化的重要经济植物,其自然资源非常丰富,亦是民间常用的香料植物资源。该研究采用性状、显微及理化鉴定方法,对艳山姜果实进行了系统的生药学研究,并对α-蒎烯、莰烯、β-蒎烯及1,8-桉叶油醇四个主要心血管药理活性成分进行含量分析。结果表明:艳山姜果实性状鉴别特征为果皮见12~20条纵棱隆起,顶端具花被残基突起,种子团由白色隔膜分为3瓣,每瓣具种子8~20粒不等,较易散落。显微鉴别特征为:种子横切面具1~2列油细胞,外胚乳细胞含淀粉粒,并可见细小草酸钙方晶;果实粉末可见螺纹导管、草酸钙方晶、淀粉粒、石细胞等。气相色谱法测得艳山姜果实挥发油中α-蒎烯、莰烯、β-蒎烯及1,8-桉叶油醇的平均含量分别为4.292%、3.966%、9.703%、27.171%。该研究的性状、显微鉴别方法准确、简单、易行,可作为艳山姜药材的鉴别依据;气相色谱含量测定方法重现性好,测定结果精确可靠,可用于艳山姜果实挥发油的含量测定。该研究结果为艳山姜药材的鉴定及进一步开发利用提供科学参考。     

CRIM1 complexes with ß-catenin and cadherins, stabilizes cell-cell junctions and is critical for neural morphogenesis

In multicellular organisms, morphogenesis is a highly coordinated process that requires dynamically regulated adhesion between cells. An excellent example of cellular morphogenesis is the formation of the neural tube from the flattened epithelium of the neural plate. Cysteine-rich motor neuron protein 1 (CRIM1) is a single-pass (type 1) transmembrane protein that is expressed in neural structures beginning at the neural plate stage. In the frog Xenopus laevis, loss of function studies using CRIM1 antisense morpholino oligonucleotides resulted in a failure of neural development. The CRIM1 knockdown phenotype was, in some cases, mild and resulted in perturbed neural fold morphogenesis. In severely affected embryos there was a dramatic failure of cell adhesion in the neural plate and complete absence of neural structures subsequently. Investigation of the mechanism of CRIM1 function revealed that it can form complexes with ?-catenin and cadherins, albeit indirectly, via the cytosolic domain. Consistent with this, CRIM1 knockdown resulted in diminished levels of cadherins and ?-catenin in junctional complexes in the neural plate. We conclude that CRIM1 is critical for cell-cell adhesion during neural development because it is required for the function of cadherin-dependent junctions.     

春兰ב寒香梅’试管开花诱导及其生理基础

以春兰×寒香梅杂交种(Cymbidium goeringii×Cymbidium ‘Han Xiang Mei’)为材料,MS+琼脂4 g·L-1+蔗糖20 g·L-1+椰汁100 mL·L-1为基础培养基,通过单因素试验,探讨细胞分裂素(TDZ/6-BA)和无机盐浓度(P、K)对其试管花诱导的影响,测定花芽诱导的蛋白质、可溶性总糖含量与超氧化物歧化酶(SOD)活性及激素水平(IAA、ABA)。结果表明,添加0.2 mg·L-1 TDZ和2 mg·L-1 6-BA的花芽诱导率最高,分别为14.33%和14.00%;无机盐浓度3P/3K和3P/5K的培养基花芽诱导率较高,分别达16.67%和11.33%;花芽诱导最佳培养基为MS(3P, 3K)+TDZ 0.2 mg·L-1+椰汁100 mL·L-1+琼脂 4 g·L-1+蔗糖20 g·L-1,花芽诱导率可达34%左右。生理生化指标检测显示,可溶性蛋白、可溶性总糖含量及SOD活性与花芽诱导率呈正相关;稳定的内源激素IAA和ABA含量对花芽诱导有一定的积极作用,含量过高对花芽诱导有抑制作用。     

Upregulation of KCNQ1/KCNE1 K+ channels by Klotho

Klotho is a transmembrane protein expressed primarily in kidney, parathyroid gland, and choroid plexus. The extracellular domain could be cleaved off and released into the systemic circulation. Klotho is in part effective as β-glucuronidase regulating protein stability in the cell membrane. Klotho is a major determinant of aging and life span. Overexpression of Klotho increases and Klotho deficiency decreases life span. Klotho deficiency may further result in hearing loss and cardiac arrhythmia. The present study explored whether Klotho modifies activity and protein abundance of KCNQ1/KCNE1, a K⁺ channel required for proper hearing and cardiac repolarization. To this end, cRNA encoding KCNQ1/KCNE1 was injected in Xenopus oocytes with or without additional injection of cRNA encoding Klotho. KCNQ1/KCNE1 expressing oocytes were treated with human recombinant Klotho protein (30 ng/ml) for 24 h. Moreover, oocytes which express both KCNQ1/KCNE1 and Klotho were treated with 10 µM DSAL (D-saccharic acid-1,4-lactone), a β-glucuronidase inhibitor. The KCNQ1/KCNE1 depolarization-induced current (I_Ks) was determined utilizing dual electrode voltage clamp, while KCNQ1/KCNE1 protein abundance in the cell membrane was visualized utilizing specific antibody binding and quantified by chemiluminescence. KCNQ1/KCNE1 channel activity and KCNQ1/KCNE1 protein abundance were upregulated by coexpression of Klotho. The effect was mimicked by treatment with human recombinant Klotho protein (30 ng/ml) and inhibited by DSAL (10 µM). In conclusion, Klotho upregulates KCNQ1/KCNE1 channel activity by 'mainly' enhancing channel protein abundance in the plasma cell membrane, an effect at least partially mediated through the β-glucuronidase activity of Klotho protein.