草本凋落物与尿素联合修复对油污土壤生物化学性质的影响

为研究植物残体配合施氮对石油污染土壤生物学和化学性质的综合修复能力,以紫花苜蓿(Medicago sativa)、铁杆蒿(Artemisia gmelinii)和小冠花(Coronilla varia)3种广泛分布于陕北石油污染区的草本植物凋落物为对象,分别在配合施氮调节土壤C∶N为25∶1和不配合施氮的条件下,将其与45.37 g/kg的重度石油污染土壤混合,在20—25℃、恒湿条件下进行为期180 d的室内修复试验,检测上述处理对油污土壤微生物数量、11种土壤水解酶和氧化还原酶活性以及速效N、P和K含量的影响。结果表明:(1)3种凋落物处理均可显著提高污染土壤中放线菌和真菌数量,蔗糖酶、淀粉酶、纤维素酶、木聚糖酶、脲酶、蛋白酶、磷酸酶、多酚氧化酶、过氧化氢酶和过氧化物酶活性以及速效养分含量,并显著降低土壤总石油烃含量。(2)配合施氮总体上显著强化了凋落物对污染土壤生化性质的修复作用,但对凋落物处理下木聚糖酶、脲酶、蛋白酶和脱氢酶活性和微生物数量的恢复可能产生不利影响。(3)单纯使用凋落物作为调理剂可以更为全面的修复油污土壤受损生化性质,具有高N和P含量、较低C/N、C/P比以及较低多酚和木质素含量的凋落物修复效果更好。在急需迅速修复土壤的条件下,配合适量施氮可作为强化凋落物修复效果的可选途径,但应注意其导致的部分土壤生化指标修复效果的降低。     

干旱胁迫下腐植酸对燕麦叶片非结构性碳水化合物代谢的影响

以燕麦品种‘燕科2号’为试验材料,采用盆栽方式,分别在正常供水(75%田间持水量)、中度干旱胁迫(60%田间持水量)和重度干旱胁迫(45%田间持水量)3个水分条件下喷施腐植酸(HA)和等量清水(CK),对燕麦叶片中非结构性碳水化合物(NSC)含量及相关酶活性和籽粒产量进行测定,以明确腐植酸在干旱胁迫下对燕麦叶片非结构性碳水化合物代谢变化的影响,探讨HA对燕麦耐旱性的影响及其作用机制。结果表明：(1)随着土壤水分含量的减少,燕麦叶片中蔗糖和淀粉含量逐渐显著降低,蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)活性显著降低,而酸性转化酶(S AI)和淀粉水解酶(α GC)活性显著提高。(2)燕麦叶片可溶性总糖和还原糖含量随着土壤水分含量的减少表现出先升高后降低的变化趋势,导致籽粒产量显著下降,且干旱胁迫程度越重变化幅度越大。(3)叶面喷施HA能不同程度提升中度和重度干旱胁迫下燕麦叶片中上述非结构性碳水化合物含量,并调节相关酶活性,显著提高籽粒产量,并在重度胁迫下的效果更佳。研究发现,腐植酸可以通过调控燕麦叶片NSC的代谢来响应干旱胁迫,降低叶片细胞渗透势,有效缓解干旱胁迫造成的损伤,增强植株耐旱性。     

动态心电图对睡眠呼吸暂停综合征的诊断价值及与心律失常的关系分析

目的:评估动态心电图对睡眠呼吸暂停综合征(SAS)的诊断价值并分析其与心律失常的关系。方法:选取2017年2月～2019年2月我院收治的SAS患者80例记为病变组,另取同期于我院进行体检的健康人员80例记为对照组。两组均进行动态心电图检查,并以多导睡眠监测仪检查结果为金标准,分析动态心电图诊断SAS的灵敏度、特异度以及准确度。比较两组动态心电图监测结果,心律失常以及ST-T缺血性改变发生情况。结果:动态心电图诊断SAS的灵敏度、特异度、准确度分别为92.31%、75.00%、90.00%。病变组最高心率、房性早搏次数、室性早搏次数均高于对照组,而最低心率低于对照组(均P0.05)。病变组心律失常及ST-T缺血性改变发生率均高于对照组(均P0.05)。结论:动态心电图对SAS的诊断价值较高,具有良好的灵敏度、特异度。临床工作中可通过动态心电图监测,从而及时检出心律失常及ST-T缺血性改变,进一步有效预防猝死的发生。     

黄芪甲苷Ⅳ通过影响β-actin对内皮型一氧化氮合酶的调节作用

目的：研究黄芪甲苷Ⅳ（AS-Ⅳ）对体外培养脐静脉内皮细胞中内皮型一氧化氮合酶（eNOS）的调节作用及可能的机制。方法：培养人脐静脉内皮细胞系EA-Hy926,用AS-Ⅳ进行干预,同时给予或不给予骨架蛋白β—actin聚合稳定剂phalloidin,用免疫共沉淀方法检测eNOS与单体8-actin结合状态的变化,用L-3H．精氨酸转化为L-SH-瓜氨酸的同位素法测定eNOS活性,I^125环-磷酸鸟苷（cGMP）放射免疫法检测细胞内cGMP水平,Westernblotting方法检测细胞中eNOS和蛋白激酶B（Akt）磷酸化水平,总蛋白水平。结果：（1）AS-IV作用10min后,细胞内单体β—actin与eNOS的结合明显增加（P〈0．05或P〈0．01）,预先给予phalloidin显著抑制了AS．IV引起的两者结合的增加（P〈0．01）。②AS—IV明显增加了eNOS活性（P〈0．05）、cGMP含量（P〈0．01）、eNOSSer-1177磷酸化水平（P〈0．01）、AktSer-473磷酸化水平（P〈0．001）,预先给予phalloidin明显降低了AS—IV引起的eNOS活性（P〈0．05）、cGMP含量（P〈0．01）和磷酸化水平的增加（P〈0．01）,但对Akt的磷酸化没有影响。结论：单体β—actin与eNOS的结合在AS-IV激活eNOS的过程中起着不可或缺作用,其主要是通过促进Akt对eNOSSer—1177的磷酸化来实现的。     

基孔肯雅病毒囊膜蛋白假病毒模型的构建

目的:以HIV为骨架构建单次复制性的含基孔肯雅病毒囊膜蛋白的假病毒模型,观察其对哺乳动物细胞的侵染性。方法:PCR合成基孔肯雅病毒囊膜蛋白基因,克隆到真核表达载体上,与HIV慢病毒包装系统质粒共转染293FT细胞,48 h后收培养上清,在8μg/mL Polybrene存在下感染293FT细胞,感染48 h后在荧光显微镜下观察结果。结果:PCR合成了基孔肯雅病毒囊膜蛋白基因并克隆到真核表达载体上,测序结果正确;共转染293FT细胞后,检测到基孔肯雅病毒囊膜蛋白的表达并包装成假病毒,感染新鲜293FT细胞后能够检测到绿色荧光蛋白。结论:合成的基孔肯雅病毒囊膜蛋白基因能正确表达并包装成假病毒,含基孔肯雅病毒囊膜蛋白的假病毒能感染293FT细胞并表达绿色荧光蛋白,可用该假病毒模型进一步研究基孔肯雅病毒的感染性,筛选评价抗基孔肯雅病毒药物。     

热激蛋白70的研究进展

热激蛋白(HSP70)作为分子伴侣参与细胞内许多重要反应,从而对生物体起着重要的作用。随着研究的深入,其生物学功能不断被发现和利用的同时,HSP70的应用前景也变得越来越广泛。已有研究者对HSP70的生物学功能做了详细介绍,我们主要对近年来HSP70在医学及环境监测等方面的应用进行综述。     

Association between statin therapy and outcomes in critically ill patients: a nested cohort study

Background

The effect of statin therapy on mortality in critically ill patients is controversial, with some studies suggesting a benefit and others suggesting no benefit or even potential harm. The objective of this study was to evaluate the association between statin therapy during intensive care unit (ICU) admission and all-cause mortality in critically ill patients.

Methods

This was a nested cohort study within two randomised controlled trials conducted in a tertiary care ICU. All 763 patients who participated in the two trials were included in this study. Of these, 107 patients (14%) received statins during their ICU stay. The primary endpoint was all-cause ICU and hospital mortality. Secondary endpoints included the development of sepsis and severe sepsis during the ICU stay, the ICU length of stay, the hospital length of stay, and the duration of mechanical ventilation. Multivariate logistic regression was used to adjust for clinically and statistically relevant variables.

Results

Statin therapy was associated with a reduction in hospital mortality (adjusted odds ratio [aOR] = 0.60, 95% confidence interval [CI] 0.36-0.99). Statin therapy was associated with lower hospital mortality in the following groups: patients >58 years of age (aOR = 0.58, 95% CI 0.35-0.97), those with an acute physiology and chronic health evaluation (APACHE II) score >22 (aOR = 0.54, 95% CI 0.31-0.96), diabetic patients (aOR = 0.52, 95% CI 0.30-0.90), patients on vasopressor therapy (aOR = 0.53, 95% CI 0.29-0.97), those admitted with severe sepsis (aOR = 0.22, 95% CI 0.07-0.66), patients with creatinine ≤100 μmol/L (aOR = 0.14, 95% CI 0.04-0.51), and patients with GCS ≤9 (aOR = 0.34, 95% CI 0.17-0.71). When stratified by statin dose, the mortality reduction was mainly observed with statin equipotent doses ≥40 mg of simvastatin (aOR = 0.53, 95% CI 0.28-1.00). Mortality reduction was observed with simvastatin (aOR = 0.37, 95% CI 0.17-0.81) but not with atorvastatin (aOR = 0.80, 95% CI 0.84-1.46). Statin therapy was not associated with a difference in any of the secondary outcomes.

Conclusion

Statin therapy during ICU stay was associated with a reduction in all-cause hospital mortality. This association was especially noted in high-risk subgroups. This potential benefit needs to be validated in a randomised, controlled trial.     

A comparative study on the activity and antigenicity of truncated and full-length forms of streptokinase

Application of streptokinase (SK) as a common and cost-effective thrombolytic drug is limited by its antigenicity and undesired hemorrhagic effects. Prior structural/functional and epitope-mapping studies on SK suggested that removal of 59 N-terminal residues led to its fibrin dependency and identified SK antigenic regions, respectively. Following in silico analyses two truncated SK proteins were designed and compared for their fibrin specificity and antigenicity with the full-length SK. Computer-based modeling was used to predict the effect of vector (pET41a)-born protein tags on the conformation of SK fragments. SK60-386, SK143-386 and full-length SK (1-414) were separately cloned, expressed in BL21 E. coli cells and confirmed by Western-blotting. Functional activity of the purified proteins was evaluated with chromogenic and clot lysis assays and their antigenicity was tested by ELISA assay using rabbit anti-streptokinase antibody. As expected, chromogenic bioassay showed a major activity decline for SK60-386 and SK143-386 (83 and 91 percent, respectively), compared to SK1-414. However, in clot lysis assay, which is a fibrin-dependent pharmacopoeia-approved test, SK60-386 and SK143-386 were respectively 35 and 31 percent more active though lysed the clots slower than full-length SK. Antigenic analysis also indicated significant decrease in ELISA signals obtained for truncated proteins compared to SK1-414 (45 and 28 percent less reactivity for SK143-386 and SK60-386, respectively, p < 0.0001). The results of this study for the first time pointed to SK143-386 and SK60-386, as improved SK derivatives with increased fibrin-selectivity and decreased antigenicity as well as acceptable bioactivity profiles in a pharmacopoeia-based analysis, which deserve more detailed pharmacological studies.     

转mapk双链RNA干扰表达载体黄瓜对根际土壤细菌多样性的影响

随着RNA干扰技术的发展,通过植物表达病原物特异的dsRNA来防治植物病害的转基因作物越来越多.转根结线虫mapk双链RNA表达载体的黄瓜能够通过RNA干扰作用沉默线虫的mapk基因,对根结线虫具有良好的防治效果.为了评价该转基因黄瓜的安全性,明确mapk双链RNA干扰表达载体转基因黄瓜植株对根际土壤细菌多样性的影响,采用16S rRNA基因克隆文库方法对非转基因黄瓜和转基因黄瓜土壤细菌群落多样性进行分析.结果表明,非转基因黄瓜土壤细菌文库包含124个OTU(可操作分类单元),转基因黄瓜土壤细菌文库包含122个OTU.2个文库共同拥有的OTU为115个.2个文库都包含1 3个类群细菌,Acidobacteria、Actinobacteria、Armatimonadetes、Bacteroidetes、candidate division BRC1、Chloroflexi、Firmicutes、Gemmatimonadetes、Nitrospira、Planctomycetes、Proteobacteria、Verrucomicrobia、unclassified_Bacteria. 其中 Proteobacteria、Bacteroidetes 、Chloroflexi和Acidobacteria是优势菌群.其他细菌类群数量相对较少.在纲分类水平上,两个文库包含的细菌类群一致,且各类群细菌比例差异不大.在Acidobacteria门中,Acidobacteria_Gp6为优势菌群.在Bacteroidetes门中,Sphingobacteria纲细菌数量最多.在Chloroflexi门中,unclassified Chloroflexi细菌最多.在Proteobacteria门中,其中Betaproteobacteria纲的细菌数量最多.从多样性指数角度分析,两种土壤细菌群落的Shannon、Simpson和Chao值差异不大.总体来看,两种土壤细菌类群差异不显著,转基因黄瓜未对根际土壤细菌群落产生明显影响.