Determining the involvement of two aminopeptidase Ns in the resistance of Plutella xylostella to the Bt toxin Cry1Ac: cloning and study of in vitro function

The cloning, expression in vitro, and characterization of two aminopeptidase Ns (APN5s and APN2s) isolated from the midgut of Cry1Ac-resistant (R) and susceptible (S) strains of Plutella xylostella larvae are presented in this paper. The deduced amino acid sequences of APN5s included C-terminal GPI-modification sites, the gluzincin aminopeptidase motif GATEN, and three N-glycosylated sites; those of APN2s had no GPI-modification sites, had gluzincin aminopeptidase motif GAMEN, and had four N-glycosylated sites. O-glycosylated sites were not predicted for either APN. Because APN2R and APN2S cDNAs contained the same nucleotides, only full-length cDNAs encoding APN5R and APN5S were expressed in Trichoplusia ni cells. Far-Western blotting showed that the expressed receptor APN5 bound to the Cry1Ac toxin. An enzyme-specific activity experiment also showed that APN5 genes were expressed in T. ni cells. ELISA revealed no differences in the binding of expression proteins from the resistant and susceptible strain with Cry1Ac.     

不同细胞周期大鼠肝实质细胞癌细胞粘弹特性研究

以胸腺嘧啶核苷和秋水仙碱顺序阻断法及胸腺嘧啶核苷双阻断法分别获得同步化Ｇ１期和Ｓ期细胞,从细胞周期角度出发,采用微管吸吮技术对大鼠肝实质细胞癌细胞的粘弹特性进行了测定并以标准线性固体模型对实验数据进行了拟合,结果表明：该细胞具有高弹性和低粘性的总体特征;Ｇ１期细胞与Ｓ期细胞相比具有高Ｋ１值和低μ值的特点,从而显示Ｇ１期细胞比Ｓ期细胞具有更大的强度和更快的被动变形能力。这些结果不仅反映了同步化细胞存在的细胞骨架状态的周期性差异,也提示Ｇ１期细胞可能比Ｓ期细胞更适于在血流中存活和转移。     

Protein kinase C delta null mice exhibit structural alterations in articular surface,intra-articular and subchondral compartments

IntroductionStructural alterations in intra-articular and subchondral compartments are hallmarks of osteoarthritis, a degenerative disease that causes pain and disability in the aging population. Protein kinase C delta (PKC-δ) plays versatile functions in cell growth and differentiation, but its role in the articular cartilage and subchondral bone is not known.MethodsHistological analysis including alcian blue, safranin O staining and fluorochrome labeling were used to reveal structural alterations at the articular cartilage surface and bone–cartilage interface in PKC-δ knockout (KO) mice. The morphology and organization of chondrocytes were studied using confocal microscopy. Glycosaminoglycan content was studied by micromass culture of chondrocytes of PKC-δ KO mice.ResultsWe uncovered atypical structural demarcation between articular cartilage and subchondral bone of PKC-δ KO mice. Histology analyses revealed a thickening of the articular cartilage and calcified bone–cartilage interface, and decreased safranin O staining accompanied by an increase in the number of hypertrophic chondrocytes in the articular cartilage of PKC-δ KO mice. Interestingly, loss of demarcation between articular cartilage and bone was concomitant with irregular chondrocyte morphology and arrangement. Consistently, in vivo calcein labeling assay showed an increased intensity of calcein labeling in the interface of the growth plate and metaphysis in PKC-δ KO mice. Furthermore, in vitro culture of chondrocyte micromass showed a decreased alcian blue staining of chondrocyte micromass in the PKC-δ KO mice, indicative of a reduced level of glycosaminoglycan production.ConclusionsOur data imply a role for PKC-δ in the osteochondral plasticity of the interface between articular cartilage and the osteochondral junction.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-015-0720-4) contains supplementary material, which is available to authorized users.     

常用食品防腐剂抗菌作用的研究

采用杯碟法和琼脂平板点种法分别测定２０种常用食品防腐剂对３０株菌的抑菌圈直径和最低抑菌浓度,借此判断药剂抗菌力的大小。实验结果表明,在所测２０种防腐剂中,对细菌抑制力最强者是乙酸,对霉菌和酵母菌抑制力最强者是富马酸二甲酯。     

P38MAPK信号通路与心血管疾病关系的研究进展

P38MAPK信号通路是细胞内主要的信息传递途径之一,与其他信号通路相互联系,共同调节细胞增殖、分化、凋亡、细胞骨架重构及细胞周期,在多种心血管疾病发生发展和转归中均起着重要作用。通过阻断和调控P38MAPK的表达和活性,探索防治心血管疾病的新的治疗手段具有重大临床意义。本文拟将P38MAPK与心血管疾病的关系的研究进展做一综述。     

Multi-environment mapping and meta-analysis of 100-seed weight in soybean

100-Seed weight (100-SW) of soybean is an important but complicated quantitative trait to yield. This study was focus on the quantitative trait loci (QTLs) of soybean 100-SW from 2006 to 2010, using recombination inbred lines population that was derived from a cross between Charleston and Dongnong 594. A total of 23 QTLs for 100-SW were detected in the linkage group C2, D1a, F, G and O. Nine QTLs were identified by composite interval mapping including one QTL with the minimum confidence interval (CI) of 1.3?cM, while 14 QTLs by multiple interval mapping. Furthermore, 94 reported QTLs of 100-SW were integrated with our QTL mapping results using BioMercator. As a result, 15 consensus QTLs and their corresponding markers were identified. The minimum CI was reduced to 1.52?cM by the combination of meta-analysis. These findings may merit fine-mapping of these QTL in soybean.     

剩余活性污泥完全资源化利用微生物集成技术

剩余活性污泥完全资源化利用微生物集成技术包括: 使用土著PHA合成菌回注法驯化并发酵活性污泥, 生产生物降解材料聚羟基脂肪酸酯(PHA); 采用土著嗜酸性氧化亚铁硫杆菌和氧化硫硫杆菌进行生物淋滤, 去除重金属; 以解磷菌和解钾菌为菌种, 进行固态发酵, 生产生物菌肥。结果表明, 500 L中试PHA占挥发性悬浮固体的20%以上; 重金属含量达到国家排放要求; 生物菌肥中活菌数大于1×108 个/g以上。实现了剩余活性污泥的近零排放。     

Phytoextraction of heavy metals from contaminated soil by co-cropping with chelator application and assessment of associated leaching risk

Phytoextraction using hyperaccumulating plants is generally time-consuming and requires the cessation of agriculture. We coupled chelators and a co-cropping system to enhance phytoextraction rates, while allowing for agricultural production. An experiment on I m3 lysimeter beds was conducted with a co-cropping system consisting of the hyperaccumulator Sedum alfredii and low-accumulating corn (Zea Mays, cv. Huidan-4), with addition ofa mixture of chelators (MC), to assess the efficiency of chelator enhanced co-crop phytoextraction and the leaching risk caused by the chelator. The results showed that the addition of MC promoted the growth of S. alfredii in the first crop (spring-summer season) and significantly increased the metal phytoextraction. The DTPA-extractable and total metal concentrations in the topsoil were also reduced more significantly with the addition of MC compared with the control treatments. However, mono-cropped S. alfredii without MC was more suitable for maximizing S. alfredii growth and therefore phytoextraction of Zn and Cd during the autumn-winter seasons. No adverse impact to groundwater due to MC application was observed during the experiments with three crops and three MC applications. But elevated total Cd and Pb concentrations among subsoils compared to the initial subsoil concentrations were found for the co-crop + MC treatment after the third crop.     

L-天冬酰胺酶工程菌株培养条件及稳定性

L-天冬酰胺酶工程菌株的酶活和表达水平受菌体生物量和诱导时间的影响。在生物量Ａ６００03×１０左右,热诱导4h酶活力和表达水平可达到较高水平。葡萄糖对酶的生成有阻遏作用,当葡萄糖浓度大于025%时,对工程菌酶的合成造成阻遏。确定了工程菌培养的培养基、pH值、接种量等因素。重组质粒pASN在\%E.coli\% JM105,TG1和AS1357等宿主菌中具有很好的稳定性,工程菌培养50代以上重组质粒保留90%以上,在LB和M\|3培养基中也较稳定。