MEK kinase 2 and the adaptor protein Lad regulate extracellular signal-regulated kinase 5 activation by epidermal growth factor via Src

Lad is an SH2 domain-containing adaptor protein that binds MEK kinase 2 (MEKK2), a mitogen-activated protein kinase (MAPK) kinase kinase for the extracellular signal-regulated kinase 5 (ERK5) and JNK pathways. Lad and MEKK2 are in a complex in resting cells. Antisense knockdown of Lad expression and targeted gene disruption of MEKK2 expression results in loss of epidermal growth factor (EGF) and stress stimuli-induced activation of ERK5. Activation of MEKK2 and the ERK5 pathway by EGF and stress stimuli is dependent on Src kinase activity. The Lad-binding motif is encoded within amino acids 228 to 282 in the N terminus of MEKK2, and expression of this motif blocks Lad-MEKK2 interaction, resulting in inhibition of Src-dependent activation of MEKK2 and ERK5. JNK activation by EGF is similarly inhibited by loss of Lad or MEKK2 expression and by blocking the interaction of MEKK2 and Lad. Our studies demonstrate that Src kinase activity is required for ERK5 activation in response to EGF, MEKK2 expression is required for ERK5 activation by Src, Lad and MEKK2 association is required for Src activation of ERK5, and EGF and Src stimulation of ERK5-regulated MEF2-dependent promoter activity requires a functional Lad-MEKK2 signaling complex.     

乳腺癌组织TGFβ1及其受体与 NF-κB表达的研究

目的研究转化生长因子(TGF)β1及其I型受体(TGFβRI)与核转录因子(NF)κB在乳腺癌组织中的蛋白表达及其与临床病理特征的关系.方法建立40例乳腺癌标本共120个微组织的组织芯片,应用免疫组织化学S-P法检测乳腺癌组织TGFβ1、TGFβRI与 NF-κB蛋白的表达情况.结果 TGFβ1、TGFβRI和NF-κB在乳腺癌中均有表达,三者过表达率分别为80%、65%和80%;TGFβ1的表达强度与TNM分期、组织学分级、腋淋巴结受累呈正相关(P<0.05,P<0.01和P<0.01),NFκB的表达强度与组织学分级、TNM分期呈正相关(P<0.05),TGFβRI的表达与分级、TNM分期、腋淋巴结受累均无明显相关性;TGFβ1和NF-κB的表达呈显著正相关(P<0.01).结论 TGFβ1和NF-κB的过表达与乳腺癌恶性及预后有密切关系, NF-κB可能在一定程度上受到TGFβ1的调控.     

Loci and candidate gene identification for soybean resistance to Phytophthora root rot race 1 in combination with association and linkage mapping

Molecular Breeding - Northern corn leaf blight (NCLB) is one of the main diseases of maize, which greatly reduces production and causes millions of dollars in losses worldwide annually....     

Ursolic acid stimulates UCP2 expression and protects H9c2 cells from hypoxia-reoxygenation injury via p38 signaling

Oxidative stress and apoptosis is involved in hypoxia-reoxygenation (H/R) induced myocardial injury. Increased expression of uncoupling protein 2 (UCP2), a cationic carrier protein, has protective effect against H/R injury. The present study aimed to find candidate drugs for H/R induced cardiac damage by identifying compounds regulating UCP2 expression. Here, among six natural compounds, ursolic acid (UA) had the most significant induction effect on UCP2 expression in H9c2 cells under H/R conditions. Subsequently, we found that UA significantly attenuated cell apoptosis and Caspase 3 activity, but increased nitric oxide (NO) release under H/R conditions. Additionally, UA pretreatment also decreased reactive oxygen species (ROS) production and malondialdehyde (MDA) content, but increased superoxide dismutase (SOD) activity. H/R caused a notable increase in the phosphorylation of p38, which was weakened by UA pretreatment. Moreover, p38 inhibitor (SB203580) showed the similar effects on H/R cells as UA pretreatment, while UCP2 knockdown had the reverse biological effects. More importantly, the effects of UA or p38 inhibitor exposure were partially rescued by UCP2 knockdown. Collectively, our data suggested the functions of UA on UCP2 expression and on the protection of H/R-stimulated H9c2 cells may be attributed to p38 signaling pathway.     

Development of a Efficient and Sensitive Dispersive Liquid–Liquid Microextraction Technique for Extraction and Preconcentration of 10 β2-Agonists in Animal Urine

Dispersive liquid–liquid microextraction (DLLME) coupled with ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) was developed for the extraction and determination of 10 β₂-agonists in animal urine. Some experimental parameters, such as the type and volume of the extraction solvent, the concentration of the dispersant, the salt concentration, the pH value of the sample solution, the extraction time and the speed of centrifugation, were investigated and optimized. Under the optimized conditions, a good enrichment factors (4.8 to 32.3) were obtained for the extraction. The enrichment factor show that the concentration rate of DLLME is significantly higher than other pretreatment methods, and the detection sensitivity has been greatly improved. The calibration curves were linear, the correlation coefficient ranged from 0.9928 to 0.9999 for the concentration range of 0.05 to 50 ngmL^-1 and 0.1 to 50 ngmL^-1, and the relative standard deviations (RSDs, n = 15, intra and inter-day precision) at a concentration of 5 ngmL^-1 were in the range of 1.8 to 14.6%. The limits of detection (LODs) for the 10 β₂-agonists, based on a signal-to-noise ratio (S/N) of 3, were in the range of 0.01 to 0.03 ngmL^-1. The proposed method was used to identify β₂-agonists in three types of animal urine (swine, cattle, sheep), and the relative recoveries from each matrix were in the range of 89.2 to 106.8%, 90.0 to 109.8% and 89.2 to 107.2%, respectively.     

伤寒Vi多糖菌苗免疫血清的LPS抗体测定

以肠炎沙门氏菌脂多糖为抗原,用酶标法测定Ｖｉ多糖菌苗免疫血清的抗ＬＰＳ抗体。各Ｖｉ多糖菌苗组免疫后１月,６月的抗ＬＰＳ水平均显著高于免前（Ｐ＜０．０００１）,对照组无显著差异（ｐ＞０．１）。两３０μｇ菌苗组抗ＬＰＳ阳转率均约为１５％。提纯Ｖｉ多糖菌苗所含的微量伤寒ＬＰＳ,也可能为接种者提供一定的保护作用。     

A Novel Zebrafish Xenotransplantation Model for Study of Glioma Stem Cell Invasion

Invasion and metastasis of solid tumors are the major causes of death in cancer patients. Cancer stem cells (CSCs) constitute a small fraction of tumor cell population, but play a critical role in tumor invasion and metastasis. The xenograft of tumor cells in immunodeficient mice is one of commonly used in vivo models to study the invasion and metastasis of cancer cells. However, this model is time-consuming and labor intensive. Zebrafish (Danio rerio) and their transparent embryos are emerging as a promising xenograft tumor model system for studies of tumor invasion. In this study, we established a tumor invasion model by using zebrafish embryo xenografted with human glioblastoma cell line U87 and its derived cancer stem cells (CSCs). We found that CSCs-enriched from U87 cells spreaded via the vessels within zebrafish embryos and such cells displayed an extremely high level of invasiveness which was associated with the up-regulated MMP-9 by CSCs. The invasion of glioma CSCs (GSCs) in zebrafish embryos was markedly inhibited by an MMP-9 inhibitor. Thus, our zebrafish embryo model is considered a cost-effective approach tostudies of the mechanisms underlying the invasion of CSCs and suitable for high-throughput screening of novel anti-tumor invasion/metastasis agents.     

Gene identification and expression analysis of 86,136 Expressed Sequence Tags (EST) from the rice genome

Expressed Sequence Tag (EST) analysis has pioneered genome-wide gene discovery and expression profiling. In order to establish a gene expression index in the rice cultivar indica, we sequenced and analyzed 86,136 ESTs from nine rice cDNA libraries from the super hybrid cultivar LYP9 and its parental cultivars. We assembled these ESTs into 13,232 contigs and leave 8,976 singletons. Overall, 7,497 sequences were found similar to the existing sequences in GenBank and 14,711 are novel. These sequences are classified by molecular function, biological process and pathways according to the Gene Ontology. We compared our sequenced ESTs with the publicly available 95,000 ESTs from japonica, and found little sequence variation, despite the large difference between genome sequences. We then assembled the combined 173,000 rice ESTs for further analysis. Using the pooled ESTs, we compared gene expression in metabolism pathway between rice and Arabidopsis according to KEGG. We further profiled gene expression pattern     

A Special Tinea Nigra Caused by Curvularia lunata: Case Report and Literature Review

Tinea nigra is a superficial fungal infection usually caused by Hortaea werneckii (H. werneckii). We report a special case of tinea nigra in an immunocompetent child who developed a unilateral, rapidly growing pigmented lesion on her palm. Interestingly, Curvularia lunata (C. lunata) was isolated from the lesion scrapes and was identified by both morphological features and molecular biology methods. The lesion was completely cleared by topical naftifine hydrochloride and ketoconazole cream. We present—to the best of our knowledge—the first case of tinea nigra where the causative pathogen was identified as C. lunata. We therefore provide a brief literature review of previously reported cases of tinea nigra to broaden the knowledge of the potential causative pathogens. The etiology, demography, clinical features, diagnostic methods, and treatment of the reviewed cases are summarized and analyzed.