慢性阻塞性肺疾病急性加重期患者咽部菌群与免疫指标的相关性

目的探讨慢性阻塞性肺疾病(COPD)急性加重期患者咽部菌群与免疫指标的相关性,为该类患者的治疗提供参考。方法选取2016年6月至2019年8月期间于我院就诊的125例COPD患者作为研究对象,其中急性加重期60例(急性加重组),缓解期65例(缓解组)。选取同期于我院进行健康体检者60例作为对照组。比较3组对象咽部菌群检出情况和CD3~+细胞、CD4~+细胞及CD4~+/CD8~+水平,并进行Spearman相关性分析。结果急性加重组患者咽部草绿色链球菌(33.3%)、干燥奈瑟菌(25.0%)比例及菌群多样性(4.713±0.786)显著低于缓解组和对照组(均P0.05),致病菌铜绿假单胞菌、金黄色葡萄球菌、肺炎链球菌等比例及菌群总密集度均显著高于缓解组和对照组(均P0.05)。急性加重组患者CD3~+细胞、CD4~+细胞及CD4~+/CD8~+水平均显著低于缓解组和对照组(均P0.05)。Spearman相关性分析显示,COPD急性加重期患CD3~+细胞、CD4~+细胞及CD4~+/CD8~+水平与菌群总密集度均呈负相关,与菌群多样性均呈正相关(均P0.05)。结论 COPD急性加重期患者存在明显咽部菌群失衡,可引起机体免疫状态的紊乱。     

Regulation of root secondary metabolites by partial root-associated microbiotas under the shaping of licorice ecotypic differentiation in northwest China

Interactions between plant hosts and their microbiotas are becoming increasingly evident, while the effects of plant communities on microbial communities in different geographic environments are poorly understood. Here, the differentiation of licorice plant ecotypes and the distribution of root-associated microbiotas were investigated across five sampling sites in northwest China. The interactions between the environment, plant and microbial communities, and their effects on licorice root secondary metabolites, were elucidated. The plant community was clearly differentiated into distinct ecotypes based on genotyping-by-sequencing and was primarily driven by geographic distance and available soil nitrogen. The bulk soil and root-associated microbiotas (rhizosphere soil and root endosphere) partially correlated with plant community, but all were significantly discriminated by plant clade. Moreover, these microbiotas were explained to different extents by distinct combinations of environment, geography, and plant community. Similarly, three structural equation models showed that licorice root secondary metabolites were complicatedly modulated by multiple abiotic and biotic factors, and were mostly explained by these factors in the rhizosphere model. Collectively, the results provide novel insights into the role of environment–plant–microbiota interactions in regulating root secondary metabolites. That should be accounted for when selecting appropriate licorice planting sites and management measures.     

大鼠Mocs2基因跳跃型外显子可变剪接保留率的检测

可变剪接是真核生物基因表达调控的一种重要方式,它通过剪接位点调控ORFs (Open reading frames)区域外显子或内含子的表达,产生不同的mRNA剪接体,进一步翻译成具有相同或不同功能的蛋白质,从而对诸如发育、疾病、环境响应等生物学过程产生重要影响.已有文献报道,人类Mocs2基因exon 1a与exon 1b外显子发生互斥型可变剪接,产生的两个剪接体分别编码钼喋呤合酶的大小亚基,参与钼辅因子生物的合成过程.基于本课题组前期大鼠肺部组织RNA-seq测序的结果,发现Mocs2基因2号外显子转录后可被剪接或保留,产生两个不同的剪接体.为验证测序结果的准确性,本研究利用半定量PCR与实时荧光定量PCR两种方法检测大鼠脑、海马、肺组织中Mocs2基因2号外显子的保留率.半定量PCR方法检测大鼠肺、海马、大脑组织中Mocs2基因的保留率分别为(88.28±3.09)％、(99.44±0.24)％、(99.54±0.19)％.实时荧光定量PCR检测的保留率分别为(66.76±20.47)％、(75.60±12.44)％、(87.28±16.4)％.实验结果表明,Mocs2基因2号外显子在大鼠中存在可变剪接现象,且不同组织具有一定差异.本工作进一步验证了两种PCR方法检测前体mRNA可变剪接保留率的可行性.     

1株甲苯降解真菌的筛选鉴定及其降解甲苯的特性

从实验室保存的7株真菌筛选到1株能高效降解甲苯的菌株H1,基于形态特征、ITS序列系统学分析,将H1菌株鉴定为毛栓菌（Trametes hirsuta）。利用正交设计实验方法研究了温度、pH值、甲苯浓度和吐温80浓度对H1菌株降解甲苯的影响,研究得出该菌株降解甲苯的最适条件为30℃、pH 5.0、甲苯浓度300mg/L、吐温80浓度0.05%,在该条件下H1对甲苯的最大降解率为85.3%,降解率比未优化之前有了显著提高。比较了H1菌株在3种培养基产生漆酶的能力,H1在土豆葡萄糖培养基产酶能力最强,在第7天达到酶活高峰16 500 U/L。H1在甲苯为唯一碳源的培养基中,漆酶酶活最低,培养7 d时漆酶酶活为589 U/L。     

Isolation, folding and structural investigations of the amino acid transporter OEP16

Membrane proteins compose more than 30% of all proteins in the living cell. However, many membrane proteins have low abundance in the cell and cannot be isolated from natural sources in concentrations suitable for structure analysis. The overexpression, reconstitution, and stabilization of membrane proteins are complex and remain a formidable challenge in membrane protein characterization. Here we describe a novel, in vitro folding procedure for a cation-selective channel protein, the outer envelope membrane protein 16 (OEP16) of pea chloroplast, overexpressed in Escherichia coli in the form of inclusion bodies. The protein is purified and then folded with detergent on a Ni–NTA affinity column. Final concentrations of reconstituted OEP16 of up to 24 mg/ml have been achieved, which provides samples that are sufficient for structural studies by NMR and crystallography. Reconstitution of OEP16 in detergent micelles was monitored by circular dichroism, fluorescence, and NMR spectroscopy. Tryptophan fluorescence spectra of heterologous expressed OEP16 in micelles are similar to spectra of functionally active OEP16 in liposomes, which indicates folding of the membrane protein in detergent micelles. CD spectroscopy studies demonstrate a folded protein consisting primarily of α-helices. ¹⁵N-HSQC NMR spectra also provide evidence for a folded protein. We present here a convenient, effective and quantitative method to screen large numbers of conditions for optimal protein stability by using microdialysis chambers in combination with fluorescence spectroscopy. Recent collection of multidimensional NMR data at 500, 600 and 800 MHz demonstrated that the protein is suitable for structure determination by NMR and stable for weeks during data collection.     

The spatio-temporal pattern of historical disturbances of an evergreen broadleaved forest in East China: a dendroecological analysis

Evergreen broadleaved forests (EBLF), the zonal forest ecosystem of the subtropical zone in east China, have been degraded from recent anthropogenic disturbance. Understanding the role of past disturbances in EBLFs would be helpful to the restoration of degraded EBLFs. We used dendroecological techniques to reconstruct the disturbance history of a secondary EBLF dominated by Schima superba and Castanopsis carlesii in Tiantong National Forest Park (29°48′N, 121°47′E), Zhejiang Province, East China. The disturbances were inferred from tree-ring growth release and long-term establishment patterns obtained from 91 overstory trees. The initial growth rates of these trees were compared to trees originating in the understory to evaluate the intensity of past disturbances. The spatial distribution patterns of disturbances were portrayed with tree mapping. The results revealed that there were five disturbances, averaging one disturbance per decade over the past half century. The first disturbance event was probably most intense given that most canopy trees established at that time and displayed high initial growth rates. The timing of the second disturbance event coincided with the documented selective logging. The last three disturbances, having lower tree growth responses and a clumped spatial distribution of gap creation, were probably the result of recurring typhoons. The first two disturbances led to tree regeneration and secondary succession, represented mainly by long-lived deciduous trees in the forest. The subsequent disturbances facilitated the stand development process, creating a complex three-dimensional structure from a pre-existing single-age cohort. This study suggests that EBLFs affected by large disturbances can recover in a few decades and the frequent gap disturbances probably facilitate its process in the early successional stages.     

Genome sequence of the stramenopile <Emphasis Type="Italic">Blastocystis</Emphasis>, a human anaerobic parasite

Background

Blastocystis is a highly prevalent anaerobic eukaryotic parasite of humans and animals that is associated with various gastrointestinal and extraintestinal disorders. Epidemiological studies have identified different subtypes but no one subtype has been definitively correlated with disease.

Results

Here we report the 18.8 Mb genome sequence of a Blastocystis subtype 7 isolate, which is the smallest stramenopile genome sequenced to date. The genome is highly compact and contains intriguing rearrangements. Comparisons with other available stramenopile genomes (plant pathogenic oomycete and diatom genomes) revealed effector proteins potentially involved in the adaptation to the intestinal environment, which were likely acquired via horizontal gene transfer. Moreover, Blastocystis living in anaerobic conditions harbors mitochondria-like organelles. An incomplete oxidative phosphorylation chain, a partial Krebs cycle, amino acid and fatty acid metabolisms and an iron-sulfur cluster assembly are all predicted to occur in these organelles. Predicted secretory proteins possess putative activities that may alter host physiology, such as proteases, protease-inhibitors, immunophilins and glycosyltransferases. This parasite also possesses the enzymatic machinery to tolerate oxidative bursts resulting from its own metabolism or induced by the host immune system.

Conclusions

This study provides insights into the genome architecture of this unusual stramenopile. It also proposes candidate genes with which to study the physiopathology of this parasite and thus may lead to further investigations into Blastocystis-host interactions.

     

Analysis of 2‐propanol in exhaled breath using in situ enrichment and cataluminescence detection

We report a simple gaseous sensor for the sensitive detection of trace 2‐propanol in exhaled breath using in situ enrichment and cataluminescence detection method on the surface of nanomaterials. The influences of heating voltage and absorption time on the CTL intensity were discussed, respectively. In the selected conditions, the linear range of 2‐propanol concentration is 60–600 ppbv and the detection of limit is 11 ppbv. Moreover, the lifetime and selectivity of the sensor were also investigated. It has the potential to diagnostic volatile organic compounds in human breath. Copyright © 2010 John Wiley & Sons, Ltd