Improved biological characteristics of poly(L-lactic acid) electrospun membrane by incorporation of multiwalled carbon nanotubes/hydroxyapatite nanoparticles

Significant effort has been devoted to fabricating various biomaterials to satisfy specific clinical requirements. In this study, we developed a new type of guided tissue regeneration (GTR) membrane by electrospinning a suspension consisting of poly( l-lactic acid), multiwalled carbon nanotubes, and hydroxyapatite (PLLA/MWNTs/HA). MWNTs/HA nanoparticles were uniformly dispersed in the membranes, and the degradation characteristics were far improved. Cytologic research revealed that the PLLA/MWNTs/HA membrane enhanced the adhesion and proliferation of periodontal ligament cells (PDLCs) by 30% and inhibited the adhesion and proliferation of gingival epithelial cells by 30% also, compared with the control group. After PDLCs were seeded into the PLLA/MWNTs/HA membrane, cell/membrane composites were implanted into the leg muscle pouches of immunodeficient mice. Histologic examinations showed that PDLCs attached on the membranes functioned well in vivo. This new type of membrane shows excellent dual biological functions and satisfied the requirement of the GTR technique successfully in spite of a monolayer structure. Compared with other GTR membranes on sale or in research, the membrane can simplify the manufacturing process, reduce the fabrication cost, and avoid possible mistakes in clinical application. Moreover, it does not need to be taken out after surgery. PLLA/MWNTs/HA membranes have shown great potential for GTR and tissue engineering.     

转录因子cpcR同源基因cpcR-c1和cpcR-t的功能鉴定

苏云金芽胞杆菌（Bacillus thuringiensis,Bt） LM1212菌株与典型的Bt菌株表型不同,可分化形成芽胞、形成细胞和晶体产生细胞。在LM1212菌株中,转录因子CpcR不仅参与了细胞分化过程,而且能够激活晶体蛋白基因cry35-like的启动子(P₃₅)。【目的】筛选cpcR同源基因,验证其生物学功能。【方法】本研究克隆了2个cpcR同源基因,来源于蜡样芽胞杆菌的cpcR-c1和来源于东洋芽胞杆菌的cpcR-t,将cpcR及其同源基因分别构建在pHT304-P₃₅-gfp、pHT304-P₃₅-lacZ报告载体上,获得的重组质粒转入无cpcR基因且无晶体蛋白基因的Bt HD73^–菌株中。利用激光共聚焦显微镜观察重组菌HD^–(cpcR-c1-P₃₅-gfp)和HD^–(cpcR-t-P₃₅-gfp)的细胞表型并进行芽胞计数实验。测定HD^–(cpcR-c1-P_35<...     

一株具有防病促生功能的贝莱斯芽孢杆菌SF327

【目的】筛选植物根际促生贝莱斯芽孢杆菌,分析菌株的生防潜力和全基因组特征。【方法】通过温室小青菜促生试验以及植物益生表型的分析,明确具有促生功能的菌株SF327。用滤纸片法测定菌株SF327对5种植物病原真菌以及4种植物病原细菌的拮抗活性。通过大田喷雾接种的方式评价菌株SF327对水稻白叶枯病的防治潜力。利用antiSMASH分析预测菌株SF327产生的二次代谢产物。通过比较基因组分析SF327与2株植物根际益生贝莱斯芽孢杆菌的代表性菌株FZB42和SQR9的亲缘关系、核心基因以及二次代谢产物合成基因簇。【结果】菌株SF327能够产生生长素吲哚-3-乙酸,是一株有益的根围促生菌;对稻瘟病菌、黄瓜枯萎病菌、辣椒疫霉菌、橡胶树胶孢炭疽菌、尖孢炭疽病菌都具有明显的拮抗作用;也具有防治水稻白叶枯病的生防潜力。菌株SF327基因组全长4.08 Mb,GC含量为46.49%,共编码4 033个基因,含有13个潜在的次生代谢产物编码基因簇,不含有质粒。SF327与FZB42和SQR9具有较近的亲缘关系,有87%以上的核心基因相同,但与SQR9的亲缘关系较近。【结论】B. velezensis SF327是一株具有宽广拮抗谱的多功能菌株,具有较好的生防应用潜力。     

基于比较基因组学的高地芽孢杆菌6ww6分子标识筛选及其快速鉴定

【目的】高地芽孢杆菌(Bacillus altitudinis) 6ww6是一株根际耐盐促生菌,可以作为微生物肥料的优选菌种。为了加强菌种知识产权保护,建立菌株快速检测方法。【方法】本研究通过基因组比对分析、TBLASTn检索、NR库检索验证并剔除有其他同源的结果序列,最终得到菌株6ww6的孤儿基因,设计引物进行PCR鉴定。【结果】筛选出5个特异性基因J939_13195、J9319_05960、J9319_13355、J9319_05965和J9319_13350。引物5960F和5960R可以单一性扩增出菌株6ww6的目的基因J9319_05960,确定其为菌株6ww6的特异性分子标识。【结论】本研究确定了菌株6ww6的唯一性身份编码,建立了以比较基因组学和孤儿基因为基础的菌株水平上的鉴定方法。该方法具有快速、精确、能鉴定到菌株水平的优点,对于有价值微生物的知识产权保护提供了有力的抓手。     

CRISPR Cas9- and Cas12a-mediated gusA editing in transgenic blueberry

To develop an effective genome editing tool for blueberry breeding, CRISPR-Cas9 and CRISPR-Cas12a were evaluated for their editing efficiencies of a marker gene, beta-glucuronidase (gusA), which was previously introduced into two blueberry cultivars each a single-copy transgene. Four expression vectors were built, with CRISPR-Cas9 and CRISPR-Cas12a each driven by a 35S promoter or AtUbi promoter. Each vector contained two editing sites in the gusA. These four vectors were respectively transformed into the leaf explants of transgenic gusA blueberry and the resulting transgenic calli were induced under hygromycin selection. GUS staining showed that some small proportions of the hygromycin-resistant calli had non-GUS stained sectors, suggesting some possible occurrences of gusA editing. We sequenced GUS amplicons spanning the two editing sites in three blueberry tissues and found about 5.5% amplicons having editing features from the calli transformed with the 35S-Cas9 vector. Further, we conducted a second round of shoot regeneration from leaf explants derived from the initial Cas9- and Cas12a-containing calli (T₀) and analyzed amplicons of the target editing region. Of the newly induced shoots, 15.5% for the 35S-Cas9 and 5.3% for the AtUbi-Cas9 showed non-GUS staining, whereas all of the shoots containing the Cas12a vectors showed blue staining. Sanger sequencing confirmed the editing-induced mutations in two representative non-GUS staining lines. Clearly, the second round of regeneration had enriched editing events and enhanced the production of edited shoots. The results and protocol described will be helpful to facilitating high-precision breeding of blueberries using CRISPR Cas technologies.

     

柑橘凤蝶蛹发声器的显微结构及防御作用(英文)

【目的】动物界存在多种用于对抗捕食者的防御机制。一些鳞翅目昆虫的蛹在受到机械刺激时会发出蠕动的声音。【方法】在扫描电镜下对柑橘凤蝶Papilio xuthus蛹的发声器形态特征进行观察,并使用Audacity软件对捕获的声音进行特征分析。【结果】柑橘凤蝶蛹在腹部第4-5节和第5-6节之间的节间膜上存在发声器,有规律地发出嘶嘶声。发声器由多层甲壳素构成的刮器和板组成,刮器和板上有 50~90个突起。当蛹的腹部被寄生蜂触角刺激30 s以上时,刮器和板会迅速相互摩擦,腹部反复摆动发出声音。蛹发出的声音是由一系列每2 000~3 000 ms发生3次的短脉冲组成的。频带很宽,主要分布在5~20kHz范围内。新鲜蛹和越冬蛹的活性不同,声强也不同。【结论】本研究首先描述了柑橘蝴蝶蛹发声器的结构,结果支持了一些蝴蝶蛹对寄生生物进化出一种特殊的防御机制(声学防御)的假说。此外,通过比较两种生境的柑橘凤蝶蛹的声波特征,我们提出了同一物种在不同地理区域可以产生方言的假说。 关键词:     

Difference Analysis of ClCYC2-Like Genes Expression and DNA Methylation Between the Two Types of Florets in Chrysanthemum lavandulifolium

DNA methylation is an important epigenetic modification, that is involved in the regulation of gene expression and cell differentiation, and plays an important regulatory role in flower development in higher plants. There are two types of florets on the capitulum in the genus Chrysanthemum, the flower symmetry factor CYCLOIDEA (CYC) 2-like genes may be important candidate genes for determining the identity of the two types of florets. In this study, the diploid plant Chrysanthemum lavandulifolium was used as the research material, and qRT-PCR and bisulfite sequencing polymerase chain reaction (BSP) were used to identify the expression and DNA methylation pattern of CYC2-like genes in the two types of florets. Gene expression analysis showed that the six ClCYC2-like genes were significantly different in the two types of florets, and the expression levels of ClCYC2c, ClCYC2d, ClCYC2e and ClCYC2f in the ray florets were significantly higher than those in the disc florets. For the DNA methylation analysis of the three genes ClCYC2c, ClCYC2d, and ClCYC2e, it was found that the DNA methylation levels of these three genes were negative correlated with their expression levels, and the ways in which the three genes were regulated by the DNA methylation were different. It is speculated that the different DNA methylation of ClCYC2-like genes in the two types of florets may affect the differentiation and development of the two types of florets. This study provides new clues about epigenetics for the analysis of capitulum formation in Asteraceae.

     

Mesenchymal stem cell-derived extracellular vesicles prevent glioma by blocking M2 polarization of macrophages through a miR-744-5p/TGFB1-dependent mechanism

Cell Biology and Toxicology - Our current study is conducted with intention to explore the regulatory mechanism of mesenchymal stem cell (MSC)-derived extracellular vesicle (EV)-miR-744-5p in...     

The branch-thorn occurrence of Lycium ruthenicum is associated with leaf DNA hypermethylation in response to soil water content

Molecular Biology Reports - Lycium ruthenicum is an eco-economic shrub which can exist in two forms, thorny and thornless under varying soil moisture conditions. The aim of this study was to...