Genetic Variation in Interleukin 28B and Response to Antiviral Therapy in Patients with Dual Chronic Infection with Hepatitis B and C Viruses

Concurrent infection with hepatitis C virus (HCV) and hepatitis B virus (HBV) was not uncommon in China. To date, information on predictors of response to treatment of dually-infected HCV/HBV is limited. The aim of this study was to evaluated whether determination of the interleukin 28B (IL-28B) polymorphism statuses sufficient to predict treatment response of interferon (IFN)-based therapy in patients chronically infected with both hepatitis B and C viruses. We investigated the role of IL28B variations (rs8099917 and rs12979860) in response to IFN-based treatment and evaluated its association with the risk of the null virological response (NVR) in HCV /HBV dually-infected patients. We found that the overall distributions of the genotypes among the sustained virological response (SVR), NVR groups were significantly different (P<0.001): patients with the rs8099917 TG genotype had an increased risk of NVR (odds ratio [OR] =2.37 95% confidence interval [CI] =1.16–4.83, P =0.017), and those with the GG genotype had a further increased risk of NVR (OR=4.23, 95% CI =1.17-15.3, P=0.027). The rs12979860 allele was also highly associated with treatment failure (CT/TT vs. CC; OR =2.04, 95%CI =1.05-3.97, P =0.037). Moreover, we found that IL28B rs8099917 G variants (TG+GG) interact with HCV genotype 1(G1) to result in higher risk of NVR (P=0.009), and that they are also associated with HBV DNA reactivation (TG+GG vs. TT, P=0.005). Furthermore, multivariate regression analysis show that the rs8099917 G allele was the most important factor significantly associated with a NVR in HCV G1 patients. This study suggest that IL28B genotyping may be a valid pretreatment predictor of which patients are likely to respond to treatment in this group of difficult-to-treat HCV/HBV dually-infected patients.     

Defluorination of Aqueous Perfluorooctanesulfonate by Activated Persulfate Oxidation

Activated persulfate oxidation technologies based on sulfate radicals were first evaluated for defluorination of aqueous perfluorooctanesulfonate (PFOS). The influences of catalytic method, time, pH and K₂S₂O₈ amounts on PFOS defluorination were investigated. The intermediate products during PFOS defluorination were detected by using LC/MS/MS. The results showed that the S₂O₈ ²⁻ had weak effect on the defluorination of PFOS, while the PFOS was oxidatively defluorinated by sulfate radicals in water. The defluorination efficiency of PFOS under various treatment was followed the order: HT (hydrothermal)/K₂S₂O₈ > UV (ultraviolet)/K₂S₂O₈ > Fe²⁺/K₂S₂O₈ > US (ultrasound)/K₂S₂O₈. Low pH was favorable for the PFOS defluorination with sulfate radicals. Increase in the amount of S₂O₈ ²⁻ had positive effect on PFOS defluorination. However, further increase in amounts of S₂O₈ ²⁻ caused insignificant improvement in PFOS defluorination due to elimination of sulfate radicals under high concentration of S₂O₈ ²⁻. CF₃(CF₂)_nCOOH (n = 0–6) were detected as intermediates during PFOS defluorination. Sulfate radicals oxidation and hydrolysis were the main mechanisms involved in defluorination process of PFOS.     

不依赖天然外显子的蛋白质内含子定向进化筛选系统

蛋白质剪接技术为在蛋白质水平上直接对蛋白质进行修饰和加工提供了一种全新的解决方案,因而在蛋白质工程及相关领域具有非常广阔的应用前景。现阶段,大部分天然的蛋白质内含子在异源蛋白质中剪接活性非常低,极大限制了蛋白质内含子的开发和应用。为了开发一个可以同时对蛋白质内含子通用性和剪接活性进行筛选的系统,利用Bsa I限制性内切酶识别位点和切割不重合的特性,将Ter ThyX内含子(不含外显子序列)插入到卡那霉素抗性蛋白基因的多个位点。并且摒弃了以往需要结合天然外显子以实现剪接的方法,可以同时对蛋白质内含子的剪接活性和通用性进行筛选。Western blot结果和卡那霉素平板生长结果表明,通过卡那霉素筛选系统可以精确的将蛋白质内含子剪接反应与卡那霉素抗性结合起来,仅从卡那霉素平板上的菌落生长情况即可完成蛋白质内含子剪接活性阳性突变的筛选,是一个快速,稳定的定向进化筛选系统。     

项目式“微生物学实验”教学模式的构建与实践

微生物学实验是生物类各专业的一门重要的专业基础课程,微生物学实验技术是学生认识微生物、改造微生物和利用微生物的重要方法。课程组结合优质教学资源,以及先进仪器设备和网络技术的助力加持,构建了项目式"微生物学实验"教学模式。通过项目实验的科学设计、实施和评价,对实验教学进行了一系列的改革实践与探索,提高了实验教学效果和效率,使学生对微生物学研究有了更系统的认识,对培养学生的综合实践能力和科学研究思维起到了积极有效的促进作用。     

马铃薯立枯丝核菌拮抗菌QHZ11的实时荧光定量PCR快速检测与应用

【背景】植物根际促生菌(plant growth-promoting rhizobacteria,PGPR)在根际的定殖是其发挥作用的基础,直观有效的跟踪技术和定量方法是研究PGPR在根际原位分布规律的重要工具。【目的】建立一种马铃薯黑痣病病原菌——立枯丝核菌拮抗菌QHZ11的实时荧光定量PCR快速检测体系,并检测拮抗菌QHZ11在马铃薯根际的动态变化。【方法】根据GenBank中登录的类芽孢杆菌及近源菌株gyrB基因序列差异筛选特异性引物,优化反应条件;通过盆栽试验对马铃薯根际拮抗菌进行快速检测。盆栽试验设3个处理,T1：对照(无菌水,CK);T2：QHZ11菌悬液灌土(QHZ11);T3：将功能菌在有机肥中进行二次固体发酵制成生物有机肥(BOF11)。【结果】筛选出拮抗菌QHZ11的专用引物为gyrB-F/gyrB-R;建立的拮抗菌QHZ11实时荧光定量PCR检测方法特异性好、灵敏度高且重复性较好,线性相关系数为0.999 8,检测组内变异系数均在1%以内,扩增效率为0.9,可检测出1×103?1×1010 copies/g-soil的拮抗菌,具有检出限低和扩增效率高的特点。盆栽试验结果发现,T3处理马铃薯根际拮抗菌QHZ11的数量从接种的第10天即高出T2处理一个数量级,并于马铃薯盛花期(接种后第60天)达到峰值,说明二次固体发酵增加了拮抗菌在根际土壤中的存活和繁殖。【结论】建立的实时荧光定量PCR快速检测体系灵敏、高效,可为研究拮抗菌在马铃薯根际原位分布以及与病原菌的互作方面提供便捷有效的方法。     

云南岩溶断陷盆地土地利用方式对丛枝菌根真菌群落结构的影响

【背景】丛枝菌根真菌(arbuscular mycorrhizal fungus,AMF)是一类能与绝大部分植物形成共生关系的真菌,在植被演替和生态恢复过程中发挥重要作用。【目的】了解云南岩溶断陷盆地不同土地利用方式对AMF群落结构的影响。【方法】采用Illumina HiSeq 2500高通量测序技术对云南岩溶断陷盆地林地、灌丛和草地等3种土地利用方式土壤样品的AMF群落结构进行分析。【结果】草地总磷(total phosphorus,TP)、速效钾(available potassium,AK)、交换性镁(exchangeable magnesium,E-Mg)、电导率(electrical conductivity,EC)和土壤温度(temperature,T)显著高于林地和灌丛,林地总氮(total nitrogen,TN)和有机碳(soil organic carbon,SOC)含量显著高于灌丛和草地。TN、SOC、AK和EC是AMF群落结构最主要的影响因子。草地样品的Chao1、ACE、Shannon和Observed species指数显著高于林地和灌丛样品,尽管Simpson指数差异不显著,但是也表现出了类似的变化趋势。9个土壤样品共产生953个AMF-OTU,鉴定出AMF 3个纲4个目9个科13个属。Glomeromycetes和Paraglomeromycetes为优势纲;Glomus和Paraglomus为优势属,其次为Claroideoglomus、Acaulospora和Diversispora。Glomus和Diversispora主要分布在草地和灌丛中,随着演替的进行其相对丰度有所下降,而Paraglomus的相对丰度逐渐上升。【结论】云南岩溶断陷盆地土地利用方式的AMF群落结构具有显著差异,土壤理化因子是重要的影响因素。     

A novel peroxiredoxin from the antagonistic endophytic bacterium Enterobacter sp. V1 contributes to cotton resistance against Verticillium dahliae

Plant and Soil - [Aims] To reveal the molecular mechanism of endophytic bacteria in Verticillium wilt-resistant cottons and deeply understand the interaction between soil and plants. [Methods]...     

基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析

【目的】建立苹小卷叶蛾Adoxophyes orana转录组数据库,挖掘杀虫剂靶标及解毒代谢相关基因。【方法】采用Illumina HiSeq~(TM) 2000高通量测序技术对苹小卷叶蛾进行转录组测序,挖掘并分析杀虫剂靶标基因;利用qPCR检测6个杀虫剂靶标基因在苹小卷叶蛾卵、幼虫、蛹和成虫各不同发育阶段的表达;挖掘并分析苹小卷叶蛾转录组中解毒代谢相关基因的代谢通路及进化关系。【结果】通过组装有效序列共获得48 610条unigene(GenBank登录号:GGMW00000000)。挖掘鉴定到155个杀虫剂靶标unigene;qPCR结果显示,1个蜕皮激素受体(ecdysone receptor, ECR)、2个乙酰胆碱酯酶(acetylcholinesterase, AChE)、1个氯离子通道蛋白(chloride channel, CLC)、1个几丁质酶(chitinase, CS)和1个鱼尼丁受体(ryanodine receptor, RyR)基因在苹小卷叶蛾不同发育阶段均存在表达差异。挖掘鉴定到69个羧酸酯酶(carboxylesterase, CarE)unigene、66个谷胱甘肽S-转移酶(glutathion S-transferase, GST)unigene和205个细胞色素P450(cytochrome P450)unigene等解毒代谢相关基因,共鉴定20个CarE unigene, 32个GST unigene和30个P450 unigene与有毒物质代谢相关的通路有关。基于氨基酸序列对具有完整ORF的unigene聚类分析结果显示:12个CarEs中9个为G类,即鳞翅目保幼激素类;分别有10个AoGSTs属于Delta和Epsilon亚家族;18个P450全部聚到CYP3集团。【结论】该研究有助于苹小卷叶蛾杀虫剂靶标基因的挖掘及抗药性的研究。