巢蛋白基因沉默通过激活细胞周期依赖性激酶（cdk5）促进大鼠神经胶质瘤细胞C6的迁移和增殖

中间纤维蛋白巢蛋白(nestin)在各种胚胎前体细胞及成熟组织中均有表达.近年一些研究显示,巢蛋白的表达上调和一些恶性肿瘤的病理特征有相关性.但是,巢蛋白在干细胞分化及肿瘤发生中的作用还不为人知.在本研究中,我们运用短发卡状的RNA为工具,以大鼠神经胶质瘤细胞系C6为模型,对巢蛋白的功能进行了研究.划痕实验和迁移实验的结果均显示,巢蛋白基因沉默可以促进C6细胞的迁移.同时,BrdU渗入实验显示,此过程伴随着细胞增殖的增加.进一步研究显示,细胞周期依赖性激酶cdk5的活性在此过程中有显著的增加.此外,巢蛋白基因沉默所引起的迁移改变可以被cdk5特异性抑制剂roscovitine所回复, 而对细胞增殖则没有显著影响.综上所述,本研究揭示了巢蛋白基因沉默与神经胶质瘤细胞的迁移和增殖相关,而cdk5是此过程的重要调节因子.     

分子标记辅助改良杂交稻协优57及其父母本的蒸煮食味品质

协优57是一个产量高和适应性强的杂交中籼组合,但由于其父母本直链淀粉含量（AC）高,导致杂交稻米的AC较高、蒸煮食味品质较差。先前利用PCR-AccⅠ分子标记辅助选择对协优57的亲本057[恢复系,记作057（GG）]和协青早A[不育系,记作协A（GG）]的W x基因进行改良。利用改良前、后的各亲本分别配组,分析不同组合的AC、食味品质和颗粒性淀粉结合酶（GBSS）活性。结果表明,改良单亲的GT型组合协A（GG）×057（TT）、协A（TT）×057（GG）杂交稻米的AC由原组合协A（GG）×057（GG）的28%分别降到19.9%和19.3%,但均一性较差。改良双亲的TT纯合型组合协A（TT）×057（TT）的杂交稻米,不仅AC降到中等偏低水平（13.1%）,而且AC的均一性也有了很大的提高,蒸煮食味品质明显改善。GBSS活性分析表明：三种W x基因型的GBSS活性总体表现为GG〉GT〉TT。     

口源性口臭指示菌的筛选

目的从口源性口臭(Oral Malodor)相关菌种中筛选主要代表菌,用以建立口臭细菌学(Oral Bacteri-ology)临床辅助诊断的指示菌(Indicator bacteria)。方法用感官检测(鼻闻法)(Organoleptic test)、气相色谱(Gas Chromatography)、硫化物检测仪(Halimeter)和硫化氢检测仪(Easicult S)等4种方法,在实验室检测常见的8种牙周及龋病致病菌,通过检测鼻闻臭味程度、硫化物(Volatile sulfur compounds,VSCs)、硫化氢(Hydrogen sulfide,H2S)及有异味的短链脂肪酸(Short Chain Fatlf acid)含量来确定指示菌。结果鼻闻:牙龈卟啉单胞菌(P.gingivalis,P.g)、中间普雷沃菌(P.intermedius,P.i)和具核梭杆菌具核梭亚种(F.subsp nucleatum,F.n)恶臭明显,其他菌有微臭或无味。气相色谱检测:P.g、P.i、F.n和伴放线聚生杆菌(Aggregatibscter actinomycetemcomitans,A.a)中有异味的丁酸(Butyric acid)含量在40%～66%,其他菌,其他产物含量较低。硫化物检测:P.g、P.i和F.n的VSCs量在1 000ppb以上,硫化氢检测:P.g、P.i和F.n的H2S在600 ppb以上,其他菌两项检测均在36 ppb以下。结论 P.g、P.i和F,n是主要产臭菌种,可作为临床口臭的细菌学辅助诊断的指示菌,供临床进一步研究。     

Endoplasmic reticulum stress in wake-active neurons progresses with aging

Fragmentation of wakefulness and sleep are expected outcomes of advanced aging. We hypothesize that wake neurons develop endoplasmic reticulum dyshomeostasis with aging, in parallel with impaired wakefulness. In this series of experiments, we sought to more fully characterize age-related changes in wakefulness and then, in relevant wake neuronal populations, explore functionality and endoplasmic reticulum homeostasis. We report that old mice show greater sleep/wake transitions in the active period with markedly shortened wake periods, shortened latencies to sleep, and less wake time in the subjective day in response to a novel social encounter. Consistent with sleep/wake instability and reduced social encounter wakefulness, orexinergic and noradrenergic wake neurons in aged mice show reduced c-fos response to wakefulness and endoplasmic reticulum dyshomeostasis with increased nuclear translocation of CHOP and GADD34. We have identified an age-related unfolded protein response injury to and dysfunction of wake neurons. It is anticipated that these changes contribute to sleep/wake fragmentation and cognitive impairment in aging.     

Mapping the protein interaction network in methicillin-resistant Staphylococcus aureus

Mortality attributable to infection with methicillin-resistant Staphylococcus aureus (MRSA) has now overtaken the death rate for AIDS in the United States, and advances in research are urgently needed to address this challenge. We report the results of the systematic identification of protein-protein interactions for the hospital-acquired strain MRSA-252. Using a high-throughput pull-down strategy combined with quantitative proteomics to distinguish specific from nonspecific interactors, we identified 13,219 interactions involving 608 MRSA proteins. Consecutive analyses revealed that this protein interaction network (PIN) exhibits scale-free organization with the characteristic presence of highly connected hub proteins. When clinical and experimental antimicrobial targets were queried in the network, they were generally found to occupy peripheral positions in the PIN with relatively few interacting partners. In contrast, the hub proteins identified in this MRSA PIN that are essential for network integrity and stability have largely been overlooked as drug targets. Thus, this empirical MRSA-252 PIN provides a rich source for identifying critical proteins essential for network stability, many of which can be considered as prospective antimicrobial drug targets.     

Health and human rights in Chin State, Western Burma: a population-based assessment using multistaged household cluster sampling

Background

The Chin State of Burma (also known as Myanmar) is an isolated ethnic minority area with poor health outcomes and reports of food insecurity and human rights violations. We report on a population-based assessment of health and human rights in Chin State. We sought to quantify reported human rights violations in Chin State and associations between these reported violations and health status at the household level.

Methods and Findings

Multistaged household cluster sampling was done. Heads of household were interviewed on demographics, access to health care, health status, food insecurity, forced displacement, forced labor, and other human rights violations during the preceding 12 months. Ratios of the prevalence of household hunger comparing exposed and unexposed to each reported violation were estimated using binomial regression, and 95% confidence intervals (CIs) were constructed. Multivariate models were done to adjust for possible confounders. Overall, 91.9% of households (95% CI 89.7%–94.1%) reported forced labor in the past 12 months. Forty-three percent of households met FANTA-2 (Food and Nutrition Technical Assistance II project) definitions for moderate to severe household hunger. Common violations reported were food theft, livestock theft or killing, forced displacement, beatings and torture, detentions, disappearances, and religious and ethnic persecution. Self reporting of multiple rights abuses was independently associated with household hunger.

Conclusions

Our findings indicate widespread self-reports of human rights violations. The nature and extent of these violations may warrant investigation by the United Nations or International Criminal Court. Please see later in the article for the Editors'' Summary     

The EGFP/hERG fusion protein alter the electrophysiological properties of hERG channels in HEK293 cells

EGFP (enhanced green fluorescent protein) tagged to either the N (amino)-terminus [EGFP/hERG (human ether-a-go-go-related gene)] or C (carboxyl)-terminus (hERG/EGFP) of hERG channel is used to study mutant channel protein trafficking for several years. However, it has been reported that the process can alter hERG channel properties. The aim of the study was to determine whether EGFP tagged to N-terminus of hERG channels would alter the cellular localizations and the electrophysiological properties of hERG channels compared with untagged hERG channels. The hERG channels tagged with or without EGFP were transiently expressed in HEK (human embryonic kidney) 293 cells using a lipofectamine method. HEK 293 cells expressing pCDNA3-hERG or pEGFP-hERG were double immunolabelled with anti-hERG and anti-calnexin (an ER marker protein) followed with FITC- and TRITC (tetramethylrhodamine β-isothiocyanate)-labelled secondary antibodies, respectively. Confocal laser scanning microscope was used to observe the cellular localization of EGFP-tagged hERG channels and untagged hERG channels. Patch-clamp technique was used to record whole cell currents. We found that the EGFP/hERG fusion protein and untagged hERG channels were both expressed not only on the cell surface membrane but also in the cytoplasm of HEK293 cells. The EGFP/hERG appeared to influence the hERG channel gating properties, including reduction of the peak tail current density, more rapid inactivation process, faster recovery from inactivation and faster deactivation kinetics compared with untagged hERG channels. Our results suggest that the EGFP/hERG channel alter the electrophysiological properties of hERG channel, but it does not seem to alter the cellular location of hERG channels. Thus, EGFP tagging to N-terminus might be used for research of subcellular location of hERG channels but not for the channel electrophysiological properties.     

改良CCI大鼠痛行为学检测与损伤区自发放电活动再观察

目的：应用改良CCI模型研究外周神经损伤后痛觉过敏和自发放电各自特征及相互关系。方法：雄性SD大鼠,随机分为CCI组和Sham组,分别于术前1天和术后1、4、7、9、11、14天测定机械刺激缩足反射阈值和热缩腿反射潜伏期,同时选取术侧机械刺激缩腿反射阈值低于4g或者术侧和健侧热缩腿反射潜伏期差异大于2s的CCI模型大鼠观察术后4-14天损伤区自发放电活动。结果：神经纤维损伤后,机械痛敏和热痛敏随时间表现为逐渐增强,同时在损伤区观察到三类放电模式：整数倍放电﹑阵发放电和周期放电。结论：术后大鼠机械痛阈和热痛阈逐渐降低,机械痛敏的产生和损伤区自发放电活动关系密切,不同的放电模式可能代表不同的传入信息。     

An allosteric mechanism inferred from molecular dynamics simulations on phospholamban pentamer in lipid membranes

Phospholamban functions as a regulator of Ca(2+) concentration of cardiac muscle cells by triggering the bioactivity of sarcoplasmic reticulum Ca(2+)-ATPase. In order to understand its dynamic mechanism in the environment of bilayer surroundings, we performed long time-scale molecular dynamic simulations based on the high-resolution NMR structure of phospholamban pentamer. It was observed from the molecular dynamics trajectory analyses that the conformational transitions between the "bellflower" and "pinwheel" modes were detected for phospholamban. Particularly, the two modes became quite similar to each other after phospholamban was phosphorylated at Ser16. Based on these findings, an allosteric mechanism was proposed to elucidate the dynamic process of phospholamban interacting with Ca(2+)-ATPase.     

A human iPSC model of Hutchinson Gilford Progeria reveals vascular smooth muscle and mesenchymal stem cell defects

The segmental premature aging disease Hutchinson-Gilford Progeria syndrome (HGPS) is caused by a truncated and farnesylated form of Lamin A called progerin. HGPS affects mesenchymal lineages, including the skeletal system, dermis, and vascular smooth muscle (VSMC). To understand the underlying molecular pathology of HGPS, we derived induced pluripotent stem cells (iPSCs) from HGPS dermal fibroblasts. The iPSCs were differentiated into neural progenitors, endothelial cells, fibroblasts, VSMCs, and mesenchymal stem cells (MSCs). Progerin levels were highest in MSCs, VSMCs, and fibroblasts, in that order, with these lineages displaying increased DNA damage, nuclear abnormalities, and HGPS-VSMC accumulating numerous calponin-staining inclusion bodies. Both HGPS-MSC and -VSMC viability was compromised by stress and hypoxia in vitro and in vivo (MSC). Because MSCs reside in low oxygen niches in vivo, we propose that, in HGPS, this causes additional depletion of the MSC pool responsible for replacing differentiated cells lost to progerin toxicity.