Genistein induces G<Subscript>2</Subscript>/M cell cycle arrest via stable activation of ERK1/2 pathway in MDA-MB-231 breast cancer cells

Genistein is an isoflavonoid present in soybeans that exhibits anti-carcinogenic effect. Several studies have shown that genistein can trigger G2/M cell cycle arrest and inhibit cell growth in human breast cancer cells. In the present study, we assessed the role of MEK-ERK cascade in regulation of genistein-mediated G2/M cell cycle arrest in the hormone-independent cell line MDA-MB-231. Flow cytometric analysis showed that treatment of MDA-MB-231 cells with genistein induced a concentration-dependent accumulation of cells in the G2/M phase of the cell cycle, with a parallel depletion of the percentage of cells in G0/G1. Genistein-mediated G2/M arrest was associated with a decrease in the protein levels of Cdk1, cyclinB1, and Cdc25C as determined by Western blot analysis. Genistein induced a slow and stable activation of phosphorylated ERK1/2 in a concentration- and time-dependent manner in MDA-MB-231 cells. MEK1/2-specific inhibitor PD98059 blocked genistein-induced activation of ERK1/2 and markedly attenuated genistein-induced G2/M arrest. Furthermore, genistein induced the expression of Ras and Raf-1 protein. Genistein also up-regulated steady-state levels of both c-Jun and c-Fos. PD98059 did not depress genistein-induced up-regulation of Ras and Raf-1 protein. However, it markedly blocked genistein-induced up-regulation of c-Jun and c-Fos. These results suggest that the Ras/MAPK/AP-1 signal pathway may be involved in genistein-induced G2/M cell cycle arrest in MDA-MB-231 breast cancer cells.     

基于综合指数法的白龙江流域生物多样性空间分异特征研究

大中区域尺度上生物多样性空间分布格局识别是制定和实施区域生物多样性保护计划的前提条件,也是生物多样性保护确定优先区域研究工作迫切需要解决的关键问题之一。拟以甘肃白龙江流域为例,结合InVEST模型和遥感、GIS技术,以区域生境质量、植物净初级生产力和景观状态指数为评价指标,应用归一化处理方法,构建区域生物多样性空间格局综合评估方法,在栅格像元尺度上开展白龙江流域生物多样性评价及其空间分异特征分析。结果表明:白龙江流域生物多样性较为丰富,空间分异特征明显。生物多样性较高的区域(Ⅰ和Ⅱ级以上)面积约占39.80%,且主要集中在自然保护区和林业管护区。生物多样性较低的地区主要分布在舟曲-武都-文县的白龙江两岸及其以北区域、宕昌县岷江沿岸、高寒稀疏植被区和高山积雪-裸岩区。     

Chiral synthesis via organoboranes. 41. The utility of B-chlorodiisopinocampheylborane for a general synthesis of enantiomerically pure drugs

A general approach to the synthesis of enantiomerically pure α-phenyl amino alcohols via the asymmetric reduction of α-phenyl haloalkyl ketones or α-phenyl aminoalkyl ketones with B-chlorodiisopinocampheylborane is described. Using this approach, an improved synthesis of a potential antipsychotic, α-(4-fluorophenyl)-4-(2-pyrimidinyl)-1-piperazinebutanol in ⩾98% ee, and the broncholdilator 1-(2-methoxy-2-phenethyl)-4-(3-hydroxy-3-phenylpropyl)piperazine (eprozinol) in ⩾99% ee is achieved. © 1995 Wiley-Liss, Inc.     

A genetic demographic analysis of Lake Malawi rock‐dwelling cichlids using spatio‐temporal sampling

We estimated the effective population sizes (N_e) and tested for short‐term temporal demographic stability of populations of two Lake Malawi cichlids: Maylandia benetos, a micro‐endemic, and Maylandia zebra, a widespread species found across the lake. We sampled a total of 351 individuals, genotyped them at 13 microsatellite loci and sequenced their mitochondrial D‐loop to estimate genetic diversity, population structure, demographic history and effective population sizes. At the microsatellite loci, genetic diversity was high in all populations. Yet, genetic diversity was relatively low for the sequence data. Microsatellites yielded mean N_e estimates of 481 individuals (±99 SD) for M. benetos and between 597 (±106.3 SD) and 1524 (±483.9 SD) individuals for local populations of M. zebra. The microsatellite data indicated no deviations from mutation–drift equilibrium. Maylandia zebra was further found to be in migration–drift equilibrium. Temporal fluctuations in allele frequencies were limited across the sampling period for both species. Bayesian Skyline analyses suggested a recent expansion of M. zebra populations in line with lake‐level fluctuations, whereas the demographic history of M. benetos could only be estimated for the very recent past. Divergence time estimates placed the origin of M. benetos within the last 100 ka after the refilling of the lake and suggested that it split off the sympatric M. zebra population. Overall, our data indicate that micro‐endemics and populations in less favourable habitats have smaller N_e, indicating that drift may play an important role driving their divergence. Yet, despite small population sizes, high genetic variation can be maintained.     

地膜覆盖对北方旱地土壤水稳性团聚体及有机碳分布的影响

研究不同地膜覆盖时间对北方旱作农田土壤团聚体粒级稳定性和有机碳的影响,可为提升旱作农田生产力和保护农田环境选择合适的管理方式提供科学依据。以辽宁阜新5年秋覆膜(AP)、春覆膜(SP)和不覆膜(CK)的定位试验为研究对象,分析不同覆膜时间对0—10 cm和10—20 cm土层中2 mm、0.25—2 mm、0.053—0.25 mm和0.053 mm粒级的土壤水稳性团聚体的稳定性及有机碳的影响。结果表明,在北方旱作农田,连续5年的地膜覆盖可显著改变0—10 cm土层的土壤各级团聚体的分布、团聚体中有机碳的含量及其对土壤有机碳含量的贡献率,进而增加土壤水稳性团聚体的稳定性,而对10—20 cm土层影响不显著。与不覆膜相比,秋覆膜和春覆膜可显著提高0—10 cm土层2 mm的水稳性团聚体的含量,分别提高了36.3%、47.9%(P0.05),而对微团聚体无显著影响,说明地膜覆盖有利于提高大团聚体数量及稳定性。在0—10 cm土层,粒径2 mm团聚体有机碳含量及储量表现为秋覆膜最高,显著高于春覆膜和不覆膜处理(P0.05)。与裸地不覆膜相比,秋覆膜和春覆膜显著提高2 mm团聚体中有机碳含量对土壤有机碳的贡献率,分别提高了37%和26.1%(P0.05)。而在0—10 cm和10—20cm土层中,微团聚体中有机碳含量对土壤有机碳贡献率没有影响。在辽宁阜新土壤及种植条件下,秋覆膜处理不仅显著提高0—10 cm土壤水稳性大团聚体的含量和稳定性,还可以显著增加水稳性大团聚体有机碳含量及储量,促进有机碳的固存。     

糖化酶、木聚糖酶融合蛋白在毕赤酵母中的高效表达

通过RT-PCR方法,以埃默森篮状菌(Talaromyces emersonii)总RNA为模板,克隆出糖化酶(glucoamylase,amyA)基因的成熟肽编码序列(1 857 bp),编码618个氨基酸;以类芽孢杆菌(Paenibacillus sp.)H10-3基因组DNA为模板,克隆出木聚糖酶(xylanaseA,xynA)基因的成熟肽编码序列(636 bp),编码211个氨基酸.通过重叠延伸PCR( SOE-PCR)得到拼接片段amyA-l-xynA,并将其克隆到毕赤酵母表达载体pPIC9中,得到重组质粒pPIC9-amyA-l-xynA,重组质粒线性化后经电击转化到毕赤酵母(Pichia pastoris)GS115中,得到了表达成功的工程菌ALX2.在ALX2发酵上清液中同时检测到糖化酶活性(10.7 U/mL)和木聚糖酶活性( 51.8 U/mL).     

Comparative analysis of the gut microbiota in distinct statin response patients in East China

Statin response shows great interindividual variations. Recently, emerging studies have shown that gut microbiota is linked to therapeutic responses to drugs, including statins. However, the association between the gut bacteria composition and statin response is still unclear. In this study, gut microbiota of 202 hyperlipidemic patients with statin sensitive (SS) response and statin resistant (SR) response in East China were investigated by high throughput sequencing to compare the gut bacteria composition and biodiversity in distinct statin response patients. Higher biodiversity was detected in Group SS than Group SR. Specifically, group SS showed significantly increased proportion of genera Lactobacillus (P = 0.001), Eubacterium (P = 0.004), Faecalibacterium (P = 0.005), and Bifidobacterium (P = 0.002) and decreased proportion of genus Clostridium (P = 0.001) compared to Group SR. The results indicated that higher gut biodiversity was associated with statin sensitive response. The increased genera Lactobacillus, Eubacterium, Faecalibacterium, Bifidobacterium, and decreased genus Clostridium in patient gut microbiota may predict patient's statin response, and hence may guide statin dosage adjustments.     

PhoR/PhoP two component regulatory system affects biocontrol capability of Bacillus subtilis NCD-2

The Bacillus subtilis strain NCD-2 is an important biocontrol agent against cotton verticillium wilt and cotton sore shin in the field, which are caused by Verticillium dahliae Kleb and Rhizoctonia solani Kuhn, respectively. A mutant of strain NCD-2, designated M216, with decreased antagonism to V. dahliae and R. solani, was selected by mini-Tn10 mutagenesis and in vitro virulence screening. The inserted gene in the mutant was cloned and identified as the phoR gene, which encodes a sensor kinase in the PhoP/PhoR two-component system. Compared to the wild-type strain, the APase activities of the mutant was decreased significantly when cultured in low phosphate medium, but no obvious difference was observed when cultured in high phosphate medium. The mutant also grew more slowly on organic phosphate agar and lost its phosphatidylcholine-solubilizing ability. The suppression of cotton seedling damping-off in vivo and colonization of the rhizosphere of cotton also decreased in the mutant strain when compared with the wild type strain. All of these characteristics could be partially restored by complementation of the phoR gene in the M216 mutant.     

Overexpression of ZEB1 associated with metastasis and invasion in patients with gastric carcinoma

The aim of this study was to investigate the expression of ZEB1 in gastric carcinoma, its correlation with the clinicopathology of gastric carcinoma, and the role of ZEB1 in invasion and metastasis in gastric carcinoma. ZEB1 expression was analyzed by immunohistochemistry and Western blot in 45 gastric carcinoma tissue samples that contained the adjacent gastric mucosa. The correlation between ZEB1 expression, the occurrence and development of gastric cancer, and clinical pathology was investigated. ZEB1 expression in the human gastric carcinoma cell line AGS was downregulated by RNA interference, and changes in ZEB1 expression corresponded with changes in the invasive and metastatic ability of AGS cells. Immunohistochemistry revealed that ZEB1 protein expression in gastric carcinoma tissues was significantly higher than in normal gastric mucosa tissues (p < 0.001). A lower degree of differentiation of gastric cancer (p = 0.009), a higher TNM (tumor, node, and metastasis) stage (p = 0.010), and a larger scope of invasion were correlated with higher expression of ZEB1 (p = 0.041, 0.002). However, the expression of ZEB1 in gastric carcinoma tissue was independent of gender, age, and tumor size (p > 0.05). Western blot results also showed that ZEB1 protein expression was significantly higher in gastric carcinoma tissue than in the adjacent normal gastric mucosa tissue (p = 0.008). A lower degree of differentiation of the gastric carcinoma correlated with a higher TNM stage, and a larger scope of invasion correlated with increased ZEB1 expression (p = 0.023). Transfection of ZEB1 siRNA in AGS cells significantly decreased the expression level of ZEB1 protein (p = 0.035). Furthermore, the number of cells that could pass through the Transwell chamber was significantly lower in the transfected group than in the non-transfected control group (p = 0.039), indicating that the suppression of ZEB1 expression could significantly reduce the invasive and metastatic ability of AGS cells (p = 0.005). Concluding, in gastric carcinoma tissue, overexpression of ZEB1 may be related to the occurrence and development as well as invasion and metastasis of gastric carcinoma.