酸性土壤中高效半纤维素降解菌的筛选与鉴定

【目的】筛选能够适应南方酸性土壤的高效半纤维素降解菌株,并进行鉴定,确定菌株的安全性。【方法】采用半纤维素平板水解圈法和胞外酶测定法进行菌株筛选,通过拮抗实验构建复合微生物菌系。利用培养特征、形态学、生理生化特征和分子生物学方法进行菌株鉴定。【结果】筛选出效果稳定,互不拮抗的高效半纤维素降解放线菌4株(NA9、NA10、NA12和NA13),半纤维素酶活分别为:217.6、229.8、221.1和211.8 U/mL。真菌2株NF1和NF7,半纤维素酶活为217.7和244.2 U/mL。复合微生物菌系半纤维素酶活可达299.0 U/mL。经鉴定菌株NA9、NA10、NA12和NA13为链霉菌中的哥斯达黎加链霉菌(Streptomyces costaricanus)。菌株NF1为亮白曲霉(Aspergillus candidus),菌株NF7为黄蓝状菌(Tarlaromyces flavus)。     

Developmental regulation for collagen II gene expression in transgenic mice

In order to evaluate the involvement of the type II collagen regulatory sequences in development, we have injected a construct containing a toxin gene under the control of the rat type II collagen promoter and enhancer. The construct, pDAS10-DTA, contained the diphtheria toxin A chain gene under the control of type II collagen sequences which had been used previously to target cartilagenous tissues in transgenics. Inspection of developing fetuses at various stages of gestation revealed a high number of aborted implants as well as abnormally developing fetuses. These abnormal fetuses were of small size, had shortened and underdeveloped limbs, cleft palates, and generally resembled a phenotype similar to chondrodystrophic mice. Histological comparisons of normal and abnormal fetuses indicated a reduced amount of extracellular matrix surrounding chondrocytes, and a disorganized appearance of the tissue. These results suggest that the expression of the toxin has occurred in chondrocytes and altered the survival and development of the transgenic mice. These results also indicate that the promoter and enhancer sequences contained in the transgene controlled the developmental expression of the type II collagen gene expression.     

Intraoperative multispectral and hyperspectral label‐free imaging: A systematic review of in vivo clinical studies

Multispectral and hyperspectral imaging (HSI) are emerging optical imaging techniques with the potential to transform the way surgery is performed but it is not clear whether current systems are capable of delivering real‐time tissue characterization and surgical guidance. We conducted a systematic review of surgical in vivo label‐free multispectral and HSI systems that have been assessed intraoperatively in adult patients, published over a 10‐year period to May 2018. We analysed 14 studies including 8 different HSI systems. Current in‐vivo HSI systems generate an intraoperative tissue oxygenation map or enable tumour detection. Intraoperative tissue oxygenation measurements may help to predict those patients at risk of postoperative complications and in‐vivo intraoperative tissue characterization may be performed with high specificity and sensitivity. All systems utilized a line‐scanning or wavelength‐scanning method but the spectral range and number of spectral bands employed varied significantly between studies and according to the system's clinical aim. The time to acquire a hyperspectral cube dataset ranged between 5 and 30 seconds. No safety concerns were reported in any studies. A small number of studies have demonstrated the capabilities of intraoperative in‐vivo label‐free HSI but further work is needed to fully integrate it into the current surgical workflow.      

海洋细菌Agarivorans sp.HZ105的琼胶降解酶系

通过克隆得到菌株Agarivorans sp.HZ105中3个琼胶酶基因,长度分别为2 988 bp、1 437 bp和1 362 bp,分别编码琼胶酶HZ1、HZ3和HZ4,分别属于糖苷水解酶GH50、GH118和GH16家族。将这些琼胶酶基因与质粒p ET-32(a)构建重组表达载体,转化大肠杆菌BL21(DE3),实现了琼胶酶基因的重组原核表达,制备了重组酶,研究了琼胶酶的酶解产物。琼胶酶HZ1降解琼脂糖以及高聚合度新琼寡糖(聚合度为8、10、12和14)得到新琼二糖和新琼四糖;琼胶酶HZ3降解琼脂糖的终产物是高聚合度新琼寡糖;琼胶酶HZ4降解琼脂糖和高聚合度新琼寡糖为新琼四糖和新琼六糖。因此推测菌株HZ105主要先用琼胶酶HZ3和HZ4降解琼脂糖为较高聚合度的新琼寡糖,随后这些寡糖被琼胶酶HZ1和HZ2(课题组先前报道的另一个琼胶酶)降解为低聚合度新琼寡糖。首次研究报道了Agarivorans属中能产生4个琼胶酶的细菌菌株及其琼胶降解酶系,丰富了有关细菌降解琼胶酶体系及其中各琼胶酶作用的研究和认识,也有利于菌株HZ105琼胶酶的有效开发应用。     

Site Specific Cleavage Mediated by MMPs Regulates Function of Agrin

Background

Agrin is the key inducer of postsynaptic differentiations at the neuromuscular junction. The multidomain heparan sulfate proteoglycan is mediating via its N-terminal segment the interaction with laminin, whereas the C-terminal portion is responsible for Dystroglycan binding and clustering of the Acetylcholine receptor. Matrix metalloproteinases (MMP) are known to play essential roles in matrix remodeling, degradation and regulation of extracellular signaling networks.

Principal Findings

Site-specific processing of Agrin provides key insight into regulatory effects of Matrix metalloproteinases (MMPs). Here, we present a detailed study of agrin processing by different MMPs together with a molecular understanding of binding and cleavage at both terminal fragments. The data suggest for a regulatory effect of MMP cleavage at particularly important functional sites of agrin. Cleave of agrin abolishes the agrin-laminin complex formation and the Acetylcholine receptor clustering at the neuromuscular junction.

Conclusion/Significance

Agrin is a target of specific MMP processing resulting in agrin subfragments with different regulatory activities. MMP processing is a powerful tool to regulate extracellular signaling networks.     

FMDV非结构蛋白2C主要表位区与3AB基因的组合表达与活性分析

近年的研究表明, 口蹄疫病毒(FMDV)非结构蛋白(NSP)2C在区分灭活疫苗免疫动物与自然感染动物方面具有潜在的价值, 为了建立敏感性更高的鉴别诊断方法, 将截取了2C蛋白N-端和C-端的主要B细胞表位区和完整3AB蛋白基因组合后, 进行了原核表达, 得到了分子量约为47.6 kD的目的蛋白2C￠3AB。通过Western blotting分析证明, 表达产物能被FMDV感染动物阳性血清识别, 具有很好的反应性。以通过电泳纯化的目的蛋白作抗原进行间接ELISA检测不同来源的动物血清, 结果表明, 该抗原仅与感染动物血清具有很好的反应活性, 而与健康动物与免疫动物血清无反应, 说明重组蛋白2C￠3AB可作为区分灭活疫苗免疫动物与感染动物的鉴别诊断抗原。用2C￠3AB和3ABC为抗原进行间接ELISA, 对比检测田间血清样品, 结果显示2C￠3AB-ELISA敏感性比3ABC-ELISA高。说明以重组蛋白2C￠3AB作为鉴别诊断抗原, 能进一步提高对临界值血清的检出率, 这对区分灭活疫苗免疫动物与FMDV隐性感染动物与带毒动物具有非常重要的意义。     

Preparation and characterization of surface crosslinked TPS/PVA blend films

Surface crosslinked thermoplastic starch (TPS)/PVA blend films were prepared by applying ultra violet (UV) irradiation. Sodium benzoate was used as photosensitizer and induced onto film surface layer by soaking the TPS/PVA films in the photosensitizer aqueous solution. The effects of concentration of photosensitizer aqueous solution, soaking time and UV irradiation dose on the surface photocrosslinking reaction were investigated. Physical properties, such as water contact angle, moisture absorption, swelling degree and solubility in water as well as mechanical properties of the films were measured to characterize the influence of the surface photocrosslinking modification. The obtained results showed that the surface modification considerably reduced the surface hydrophilic character of the TPS/PVA films, enhanced the film’s water resistance and also increased tensile strength and Young’s modulus but decreased elongation at break of the films.     

链霉菌属菌株AS4.693和AS4.702的分类学研究

链霉菌属“Setae”种群原为北里孢菌属Kitasatosporia (Omura,1982)。1992年,Wellington根据16S rRNA序列分析结果将其并入链霉菌属,并建立“Setae”种群。通过对保藏的链霉菌AS 4.693、AS 4.702进行的形态学、细胞化学、分子遗传分类研究结果表明,它们与链霉菌属“Setae”种群中的典型种——西唐链霉菌Streptomyces setae(JCM3304’)具有相似性。它们的rDNA相似性高达100％,证明它们应归属于同一种群。AS.4.693定名为西唐链霉菌不规则新亚种Streptomyces setae subsp.irregularis nov.,AS 4.702定名为西唐链霉菌波曲弗氏新亚种Streptomyces setae subsp.flexuofradiae nov.。