Arsenite induces apoptosis in chinese hamster ovary cells by generation of reactive oxygen species

Arsenic, a human carcinogen, possesses a serious environmental threat but the mechanism of its toxicity remains unclear. Knowledge of how arsenic induces cell death and how cells escape the death path may help to understand arsenic carcinogenesis. We have investigated the nature of sodium arsenite-induced cell death in Chinese hamster ovary K1 cells. Following phosphate-citric acid buffer extraction, apoptotic cells with lower DNA content than the G1 cells were detected by flow cytometry. Immediately after 4 h of 40 μM arsenite treatment, no appreciable fraction of cells with sub-G1 DNA content was detected; however, the sub-G1 cell fraction increased with postarsenite incubation time, and detectable increase started at 8 h of incubation, whereas the intracellular peroxide level as measured by the fluorescent intensity of 2′,7′-dichlorofluorescein increased immediately following a 4-h arsenite treatment. Simultaneous treatment with arsenite plus antioxidant (N-acetyl-cysteine, Trolox, and Tempo); copper ion chelator (neocuproine); protein kinase inhibitor (H-7) or protein synthesis inhibitor (cycloheximide) reduced the fraction of sub-G1 cell and internucleosomal DNA degradation. Trolox, neocuproine, or cycloheximide given after arsenite treatment also effectively reduced apoptosis. These results lead to a working hypothesis that arsenite-induced apoptosis in CHO-K1 cells is triggered by the generation of hydrogen peroxide, followed by a copper-mediated Fenton reaction that catalyzes the production of hydroxyl radicals, which selectively activates protein kinase through de novo synthesis of macromolecules. © 1996 Wiley-Liss, Inc.     

喀斯特生态脆弱区典型小生境土壤团聚体稳定性比较研究

为探究喀斯特小生境类型对土壤团聚体稳定性影响,揭示小生境土壤团聚体主要失稳机制,选取研究区内石缝、石沟、石槽、石土面和土面5种典型小生境为研究对象,采用干筛法、Elliott湿筛法和Le Bissonnais法测定土壤团聚体稳定性。结果表明：湿筛后,团聚体分布以水稳性团聚体（WR_>0.25）为主,含量在81.26%以上,团聚体破坏率（PAD）介于8.93%-17.39%之间,平均重量直径（MWD）在1.94-2.85 mm间,均小于干筛,团聚体稳定性表现为石槽最好,土面最差,石土面次之。LB法处理后,MWD值变化趋势为慢速湿润（SW） > 预湿润后扰动（WS） > 快速湿润（FW）,即小生境土壤团聚体最大破坏机制为大雨或暴雨时快速湿润FW产生的消散作用,团聚体稳定性同样以石槽最好,土面最差,石土面次之。相关性分析显示,LB法测定的MWD均与湿筛法的WR_>0.25呈显著正相关,与PAD呈显著负相关,即LB法和湿筛法具有良好的相关性,LB法用于测定喀斯特地区土壤团聚体稳定性是可行的（P<0.05）。LB法FW处理的MWD值与有机质呈显著正相关,即有机质对团聚体稳定性有重要影响;全氮、全磷、全钾与WR_>0.25、SW处理的MWD值呈显著正相关,与PAD呈显著负相关,即稳定的团粒结构对养分蓄积有正向效应（P<0.05）。综合不同测定方法的结果表明,土面、石土面易造成土壤流失,应加强雨季对喀斯特生态脆弱区土面、石土面小生境的防护。     

Characterization of the Soj/Spo0J chromosome segregation proteins and identification of putative parS sequences in Helicobacter pylori

The Soj and Spo0J proteins, together with one or more parS sequences, are crucial to chromosome segregation and the progression of cell cycle in many bacteria. In Helicobacter pylori, genes coding for Soj and a plasmid replication-partition-related protein containing a Spo0J or ParB conserved domain, together with two putative parS sites identified in this study, were found to be located within the origin-proximal 20-30% of the circular chromosome. Recombinant H. pylori Spo0J bound specifically to the two putative parS sequences and that of Bacillus subtilis. In addition, hydrolysis of ATP by H. pylori Soj was accelerated in the presence of parS and/or Spo0J. Protein-protein interactions, intracellular levels, and subcellular localization of Soj and Spo0J were analyzed through polyclonal antibodies directed against recombinant Soj and Spo0J. This study was the first implication of the existence of a functional parABS system in H. pylori.     

2011-2020年长江口大型底栖动物群落结构变化趋势

对2011—2020年夏季长江口48个站位的大型底栖动物定量监测数据进行统计分析,研究长江口海域底栖生物群落时空结构演变特征。结果表明,近10年长江口海域共鉴定大型底栖动物284种,其中多毛类128种,占总种数的45.1%,甲壳类64种占22.2%,软体动物56种占19.7%,棘皮动物16种占5.6%,其他类合计20种。平均生物密度为(79.5±45.9)个/m²(年份变幅14.7—195个/m²)。平均生物量为(5.20±3.25)g/m²(年份变幅1.01—10.11g/m²),多毛类、软体动物、甲壳类是生物密度和生物量组成的主要类群。十年期间种类数、生物密度和生物量均呈现明显上升趋势,口外区贡献最突出。四个监测区域(南支、北支、杭州湾、口外)的优势种差异大。丝异须虫Heteromastus filiformis在各年份的优势种中出现的频度显著最大。总体来看,长江口监测区域大型底栖动物群落自然演变趋势向好。三项多样性指数统计结果表明,口外区大型底栖动物种类组成多样性水平显著高于口内三个区。生物群...     

牧民定居后季节草场优化配置的研究

草地畜牧业季节草场在牧民定居后新的生产力水平下应采取怎样的利用模式,是生产中亟待解决的问题。本文采用调查与实测相结合的方法,遵循草畜平衡及能物流高效持续利用原则,研究提出了新疆天山北坡中段季节草场时空优化配置模式(暖季放牧,冷季舍饲 放牧)。此模式不仅实现了草畜平衡,使生态环境得以保护和改善,且能取得显著的经济和社会效益。     

Angiopoietin-like protein 3 regulates the motility and permeability of podocytes by altering nephrin expression in vitro

It is well known that podocyte injury plays a vital role in massive proteinuria. The increase of podocyte motility results in podocyte foot process (FP) effacement, a typical form of podocyte injury. Our previous studies demonstrated that glomerular podocytes can express angiopoietin-like protein 3 (ANGPTL3) and that the increase of ANGPTL3 in dysfunctional glomerulus is correlated with podocyte FP effacement. Little is known, however, about the role of ANGPTL3 in podocyte injury. In this study, we investigated ANGPTL3’s effect on the motility and permeability of podocytes and on the expression of nephrin, a key molecule in podocytes. By scrape-wound and transwell migration assay, we found that ANGPTL3 over-expression significantly increased podocyte motility, whereas after ANGPTL3 knockdown by RNA interference, motility remained the same as that of the control group. Adriamycin (ADR) treatment significantly promoted podocyte motility. However, the same dose of ADR treatment could not promote motility after the knockdown of ANGPTL3. In addition, we assayed the diffusion of FITC-BSA across the podocytes’ monolayer to investigate whether ANGPTL3 could promote protein loss by means of an increase in podocyte motility. The results showed that the changes in the FITC-BSA permeability of the podocytes corresponded to changes in motility. Furthermore, we found that ANGPTL3 over-expression dramatically increased the expression of nephrin but that the up-regulation of nephrin induced by ADR was significantly inhibited when ANGPTL3 was diminished by RNAi. In conclusion, we found ANGPTL3 to be capable of regulating the motility and permeability of podocytes and that the mechanism of ANGPTL3’s regulation could be associated with the altered expression of nephrin.     

城市森林概念探析

在讨论、揭示有关城市森林和森林概念的基础上,对城市森林进行定义,认为城市森林是指在城市及其周边范围内以乔木为主体,达到一定的规模和覆盖度,能对周围的环境产生重要影响,并具有明显的生态价值和人文景观价值等的各种生物和非生物的综合体。进一步探讨城市森林的范围,对城市森林与园林以及城市绿地进行比较,并对城市森林的发展提出几点看法和建议。     

Restoration of p53 tumor suppressor pathway in human cervical carcinoma cells by sodium arsenite

In most cervical cancer cells, p53 and Rb are disrupted by human papillomaviruses (HPVs) E6 and E7, respectively. Restoration of p53 or Rb function by blocking E6/p53 or E7/Rb pathway might be a potential therapeutic purpose for these cancer cells. Treatment with sodium arsenite (SA) resulted in significant repression of E6 and E7 mRNA levels in SiHa cells. After E6 and E7 repression, p53 was dramatically induced and accumulated in cellular nuclei and Rb was also induced. Two p53-responsive genes, p21(waf1/cip1) and mdm2, were induced after SA treatment. Furthermore, SA also reduced the expressions of Cdc25A and cyclin B, blocked cell cycle progression at G2/M phase, and induced apoptosis in SiHa cells. SA-induced apoptosis was greatly reduced by expression of a dominant-negative mutated p53. In this study, we have first demonstrated that SA did repress E6 and E7 oncogenes, restore the p53 tumor suppressor pathway and induce apoptosis in SiHa cells. Therefore, it would be a potential strategy to promote SA as therapeutic purpose for HPV-positive cancer cells.     

Imaging protein structure in water at 2.7 nm resolution by transmission electron microscopy

We demonstrate an in situ transmission electron microscopy technique for imaging proteins in liquid water at room temperature. Liquid samples are loaded into a microfabricated environmental cell that isolates the sample from the vacuum with thin silicon nitride windows. We show that electron micrographs of acrosomal bundles in water are similar to bundles imaged in ice, and we determined the resolution to be at least 2.7 nm at doses of ～35 e/Ų. The resolution was limited by the thickness of the window and radiation damage. Surprisingly, we observed a smaller fall-off in the intensity of reflections in room-temperature water than in 98 K ice. Thus, our technique extends imaging of unstained and unlabeled macromolecular assemblies in water from the resolution of the light microscope to the nanometer resolution of the electron microscope. Our results suggest that real-time imaging of protein dynamics is conceptually feasible.     

The application of tetrahydroisoquinoline-3-carbonyl-TARGD(F)F as an anti-thrombotic agent having dual mechanisms of action

Platelet surface glycoproteins P-selectin and GPIIb/IIIa are implicated in the formation of platelet-fibrin-leukocyte thrombus and platelet-fibrin-platelet thrombus, respectively. In the current study, taking N-(3S-tetrahydroisoquinoline-3-carbonyl)-Thr-Ala-Arg-Gly-Asp-(Phe)-Phe (IQCA-TAFF) as a model compound, the molecular modeling, synthesis, and an evaluation system for a novel anti-thrombotic agent were investigated. The synthesis of IQCA-TAFF was achieved by coupling 3S-tetrahydro-isoquinoline-3-carboxylic acid (IQCA) and Thr-Ala-Arg-Gly-Asp(Phe)-Phe (TAFF). The molecular modeling indicated that IQCA-TAFF was able to occupy the active site pocket of P-selectin with its IQCA moiety and to block GPIIb/IIIa fibrinogen-binding sites with its TAFF moiety, respectively. These are consistent with the dual inhibition of the expressions of P-selectin and GPIIb/IIIa, and with the in vitro anti-platelet aggregation activity of IQCA-TAFF. Besides, the dual suppression of P-selectin and GPIIb/IIIa leads to significant in vivo efficacy of IQCA-TAFF, 500-fold higher than those of IQCA and TAFF, respectively. Transmission electron microscopy (TEM) images indicated that in water, IQCA-TAFF concentration-dependently formed nano-globes. The molecular modeling, in vitro bioassay, in vivo bioassay, action mechanism investigation, and nano-image visualization together constitute a model system to characterize the anti-thrombotic agent capable of simultaneously inhibiting P-selectin and GPIIb/IIIa mediated thrombosis.