Inhibition of coxsackievirus B3 in cell cultures and in mice by peptide-conjugated morpholino oligomers targeting the internal ribosome entry site

Coxsackievirus B3 (CVB3) is a primary cause of viral myocarditis, yet no effective therapeutic against CVB3 is available. Nucleic acid-based interventional strategies against various viruses, including CVB3, have shown promise experimentally, but limited stability and inefficient delivery in vivo remain as obstacles to their potential as therapeutics. We employed phosphorodiamidate morpholino oligomers (PMO) conjugated to a cell-penetrating arginine-rich peptide, P007 (to form PPMO), to address these issues. Eight CVB3-specific PPMO were evaluated with HeLa cells and HL-1 cardiomyocytes in culture and in a murine infection model. One of the PPMO (PPMO-6), designed to target a sequence in the 3' portion of the CVB3 internal ribosomal entry site, was found to be especially potent against CVB3. Treatment of cells with PPMO-6 prior to CVB3 infection produced an approximately 3-log(10) decrease in viral titer and largely protected cells from a virus-induced cytopathic effect. A similar antiviral effect was observed when PPMO-6 treatment began shortly after the virus infection period. A/J mice receiving intravenous administration of PPMO-6 once prior to and once after CVB3 infection showed an approximately 2-log(10)-decreased viral titer in the myocardium at 7 days postinfection and a significantly decreased level of cardiac tissue damage, compared to the controls. Thus, PPMO-6 provided potent inhibition of CVB3 amplification both in cell cultures and in vivo and appears worthy of further evaluation as a candidate for clinical development.     

Indications that live poultry markets are a major source of human H5N1 influenza virus infection in China

Human infections of H5N1 highly pathogenic avian influenza virus have continued to occur in China without corresponding outbreaks in poultry, and there is little conclusive evidence of the source of these infections. Seeking to identify the source of the human infections, we sequenced 31 H5N1 viruses isolated from humans in China (2005 to 2010). We found a number of viral genotypes, not all of which have similar known avian virus counterparts. Guided by patient questionnaire data, we also obtained environmental samples from live poultry markets and dwellings frequented by six individuals prior to disease onset (2008 and 2009). H5N1 viruses were isolated from 4 of the 6 live poultry markets sampled. In each case, the genetic sequences of the environmental and corresponding human isolates were highly similar, demonstrating a link between human infection and live poultry markets. Therefore, infection control measures in live poultry markets are likely to reduce human H5N1 infection in China.     

Suppression of proliferation and migration in highly-metastatic lung cancer cells as well as tumor growth by a new synthesized compound TBrC and its molecular mechanisms of action

To develop new anticancer agents has been considered as a useful and necessary strategy to suppress highly-metastatic lung cancer, the leading cause of cancer-related deaths in the world. In this study, we synthesized a new compound ethyl 6-bromocoumarin-3-carboxylyl L-theanine (TBrC) and studied the anticancer activity of TBrC and its molecular mechanisms of action. Our results show that TBrC remarkably inhibits the proliferation and migration in highly-metastatic lung cancer cells by inducing apoptosis and cell cycle arrest as well as regulating related protein expressions. Further study indicated that TBrC not only enhances the protein levels of Bax, cytosolic cytochrome c, caspase-3 and PARP-1 but also reduces the protein expressions of Bcl-2, cyclin D1, VEGFR1 and NF-κB as well as inhibits the phosphorylation and expressions of VEGFR2 and Akt in the cancer cells. More importantly, TBrC displays strong suppression of highly-metastatic tumor growth and reduces the tumor weight by 61.6 % in tumor-bearing mice without toxicity to the mice. Our results suggest that TBrC suppresses the proliferation and migration of lung cancer cells via VEGFR-Akt-NF-κB signaling pathways; TBrC may have a wide therapeutic and/or adjuvant therapeutic application in the treatment of lung cancer.     

Epidemiology and Outcome of Severe Sepsis and Septic Shock in Intensive Care Units in Mainland China

Introduction

Information about sepsis in mainland China remains scarce and incomplete. The purpose of this study was to describe the epidemiology and outcome of severe sepsis and septic shock in mixed ICU in mainland China, as well as the independent predictors of mortality.

Methods

We performed a 2-month prospective, observational cohort study in 22 closed multi-disciplinary intensive care units (ICUs). All admissions into those ICUs during the study period were screened and patients with severe sepsis or septic shock were included.

Results

A total of 484 patients, 37.3 per 100 ICU admissions were diagnosed with severe sepsis (n = 365) or septic shock (n = 119) according to clinical criteria and included into this study. The most frequent sites of infection were the lung and abdomen. The overall ICU and hospital mortality rates were 28.7% (n = 139) and 33.5% (n = 162), respectively. In multivariate analyses, APACHE II score (odds ratio[OR], 1.068; 95% confidential interval[CI], 1.027–1.109), presence of ARDS (OR, 2.676; 95%CI, 1.691–4.235), bloodstream infection (OR, 2.520; 95%CI, 1.142–5.564) and comorbidity of cancer (OR, 2.246; 95%CI, 1.141–4.420) were significantly associated with mortality.

Conclusions

Our results indicated that severe sepsis and septic shock were common complications in ICU patients and with high mortality in China, and can be of help to know more about severe sepsis and septic shock in China and to improve characterization and risk stratification in these patients.     

Estimating Daytime Ecosystem Respiration to Improve Estimates of Gross Primary Production of a Temperate Forest

Leaf respiration is an important component of carbon exchange in terrestrial ecosystems, and estimates of leaf respiration directly affect the accuracy of ecosystem carbon budgets. Leaf respiration is inhibited by light; therefore, gross primary production (GPP) will be overestimated if the reduction in leaf respiration by light is ignored. However, few studies have quantified GPP overestimation with respect to the degree of light inhibition in forest ecosystems. To determine the effect of light inhibition of leaf respiration on GPP estimation, we assessed the variation in leaf respiration of seedlings of the dominant tree species in an old mixed temperate forest with different photosynthetically active radiation levels using the Laisk method. Canopy respiration was estimated by combining the effect of light inhibition on leaf respiration of these species with within-canopy radiation. Leaf respiration decreased exponentially with an increase in light intensity. Canopy respiration and GPP were overestimated by approximately 20.4% and 4.6%, respectively, when leaf respiration reduction in light was ignored compared with the values obtained when light inhibition of leaf respiration was considered. This study indicates that accurate estimates of daytime ecosystem respiration are needed for the accurate evaluation of carbon budgets in temperate forests. In addition, this study provides a valuable approach to accurately estimate GPP by considering leaf respiration reduction in light in other ecosystems.     

长白山阔叶红松林降雨截留量的估算

林冠截留降雨过程是森林流域水分循环的重要组成部分,以往的研究多为1次或多次降雨量与截留量的关系,很少考虑雨强和树木特征.文中利用林冠截留降雨半经验半理论模型,以雨强和叶面积指数为模型输入,林冠湿润程度为参数,结合Penman-Monteith公式,有效地模拟了长白山阔叶红松林次降雨的截留过程和2004年5～9月的林冠截留总量.结果表明,研究期间的林冠截留总量为39.96 mm,占降雨总量的10.2%,与实测资料吻合.根据模拟结果,探讨了不同时间尺度上截留量与降雨量之间的关系,随着时间尺度的增大,截留量与降雨量的相关关系趋于明显.     

时间序列修订对森林二氧化碳通量的影响

对长白山阔叶红松林2003年生长季的涡动相关实测时间序列进行了去倾修订与超声风速仪倾斜修订,并分析了不同修订方法对森林CO2通量计算值的影响．结果表明,基于未修订时间序列计算得到的森林CO2通量(Fcraw)被高估．线性与非线性去倾对Fcraw的修订量分别为1．6％、1．8％,两者差异很小．平面拟合坐标变换与流线坐标变换对Fcraw的修订量分别为3．7％、4．7％,两者差异较大．对线性去倾后的时间序列分别进行流线坐标变换与平面拟合坐标变换,二者对Fcraw的修订量分别为5．5％与4．6％．建议对时间序列进行线性去倾与平面拟合坐标变换综合修订．     

长白山阔叶红松林CO2通量与温度的关系

应用涡度相关法观测的通量数据和环境因子数据,在生态系统水平上分析了长白山阔叶红松林生长季温度与CO2通量之间的关系.结果表明：（1）在相同的光合有效辐射水平下,净生态系统CO2交换量（NEE）随温度Ta的变化趋势为,在Ta〈20℃范围内,NEE随温度的增加而增加,在Ta=20℃附近有极大值,随温度的继续增加NEE呈下降的趋势,同时NEE还具有明显的季节变化,表现为7月〉6月〉8月〉9月〉5月〉4月〉10月.（2）应用Michaelis-Menten方程计算得出最大光合速率Pmax和生态系统呼吸Re,分析其与温度的关系发现,Pmax随温度的变化呈S型曲线,Re则随着温度的升高而呈指数上升的趋势,曲线为：Re=0.0607 exp（0.0666Tα）,R^2=0.96.夜间生态系统呼吸的Q10为3.15.（3）通过对NEE与环境因子的偏相关分析表明,温度对NEE的偏相关系数在生长季呈现先减小后增大的趋势,说明在生长季初期和末期升高温度比生长季中期对NEE的影响要大.     

科尔沁草甸生态系统水分利用效率及影响因素

生态系统水分利用效率(WUE)是衡量碳水循环耦合程度的重要指标。利用科尔沁温带草甸草地碳水通量观测数据,对该生态系统总初级生产力水分利用效率(WUEGPP)的日季变化规律及对环境和生理因子的响应进行分析。结果表明:(1)WUEGPP日变化呈下降-稳定-上升的变化趋势,最大值出现在日出后1—2 h,阴天条件下WUEGPP高于晴天,生长中期WUEGPP高于生长初期和末期;(2)总初级生产力、总蒸散和WUEGPP季节变化均呈夏季高、春秋低的形式,生长季平均值分别为0.57 mg m-2s-1、0.08 g m-2s-1和5.97 mg/g,最大值分别为1.49 mg m-2s-1、0.16 g m-2s1和13.62 mg/g;(3)总初级生产力与饱和差、气温和叶面积指数均呈二次曲线关系,与冠层导度呈对数曲线关系;总蒸散与气温呈二次曲线关系,与饱和差、叶面积指数和冠层导度相关性均不显著;(4)WUEGPP与饱和差、气温和叶面积指数均呈二次曲线关系,与冠层导度呈对数曲线关系,饱和差、冠层导度和叶面积指数分别为2.0 k Pa、0.0015 m/s和4.2是控制WUEGPP增加的阈值;(5)净生态系统生产力水分利用效率(WUENEP)和净初级生产力水分利用效率(WUENPP)季节变化规律与WUEGPP一致,均值分别为3.47和5.47 mg/g。