MV-mimicking micelles loaded with PEG-serine-ACP nanoparticles to achieve biomimetic intra/extra fibrillar mineralization of collagen in vitro

Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering.     

Cloning of aroG, the gene coding for phospho-2-keto-3-deoxy-heptonate aldolase(phe), in Escherichia coli K-12, and subcloning of the aroG promoter and operator in a promoter-detecting plasmid

Defective transducing phages carrying aroG, the structural gene for phenylalanine (phe)-inhibitable phospho-2-keto-heptonate aldolase (EC 4.1.2.15; previously known as 3-deoxy-D-arabinoheptulosonate-7-phosphate synthetase[phe]), have been isolated, and DNA from two of these phages has been used to construct a restriction map of the region from att lambda to aroG. A 7.6-kb PstI-HindIII fragment from one of these phages was cloned into pBR322 and shown to contain aroG. The location of aroG within the 7.6 kb was established by subcloning and Tn3 transpositional mutagenesis. A fragment carrying the aroG promoter and operator has been cloned into a high copy number promoter-cloning vector (pMC489), and the resulting aroGpo-LacZ' (alpha) fusion subcloned in a low copy number vector. Strains with this fusion on the low copy number vector exhibit negative regulation of beta-galactosidase expression by both phenylalanine and tryptophan and positive regulation by tyrosine in a tyrR+ background.     

TSG101 promotes the proliferation,migration and invasion of hepatocellular carcinoma cells by regulating the PEG10

The tumour susceptibility gene 101 (TSG101) is reported to play important roles in the development and progression of several human cancers. However, its potential roles and underlined mechanisms in human hepatocellular carcinoma (HCC) are still needed to be further clarified. In the present study, we reported that knock down of TSG101 suppressed the proliferation, migration and invasion of HCC cells, while overexpression of TSG101 facilitated them. Molecularly, the results revealed that knock down of TSG101 significantly decreased the cell cycle related regulatory factor p53 and p21. In another point, knock down of TSG101 also obviously decreased the level of metallopeptidase inhibitor TIMP1 (Tissue inhibitors of metalloproteinases 1), which results in inhibition of MMP2, MMP7 and MMP9. In contrast, overexpression of TSG101 had opposite effects. The iTRAQ proteomics analysis identified that oncogenic protein PEG10 (Paternally expressed gene 10) might be a potential downstream target of TSG101. Further investigation showed that TSG101 interacted with PEG10 and protected it from proteasomal degradation thereby regulating the expression of p53, p21 and MMPs. Finally, we found that both TSG101 and PEG10 proteins are up‐regulated and presented a direct correlation in HCC patients. In conclusion, these results suggest that TSG101 is up‐regulated in human HCC patients, which may accelerate the proliferation, migration and invasion of HCC cells through regulating PEG10.     

New insights into the binding mode of pyridine-3-carboxamide inhibitors of E. coli DNA gyrase

Previously we have reported on a series of pyridine-3-carboxamide inhibitors of DNA gyrase and DNA topoisomerase IV that were designed using a computational de novo design approach and which showed promising antibacterial properties. Herein we describe the synthesis of additional examples from this series aimed specifically at DNA gyrase, along with crystal structures confirming the predicted mode of binding and in vitro ADME data which describe the drug-likeness of these compounds.     

PEG模拟干旱条件下木麻黄幼苗的生理变化

以木麻黄Casuarina equisetifolia15个家系品种为材料,利用聚乙二醇6000(PEG-6000)模拟水分胁迫环境,设置三个胁迫强度处理(轻度、中度、重度胁迫),研究受胁迫2月龄木麻黄幼苗丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、脯氨酸含量的变化。结果表明,经干旱胁迫处理后,各家系幼苗叶片MDA含量随胁迫强度增大而增大,但增幅差异较大;不同干旱胁迫水平下,各家系SOD活性存在一定差异;重度胁迫条件下,除粤杂交家系外,其余家系脯氨酸含量与对照相比均为显著增加。采用主成分分析法进行抗旱性综合评价,表明MDA含量是反映水分胁迫最重要的指标。各指标综合分析结果表明,家系701、莆20、抗3、惠2、南山7和东山2抗旱性能力较好;东山短、601、59、A13抗旱能力一般;千头、粤杂交、抗1、龙4和平5抗旱能力较差。     

4种阔叶幼苗对PEG模拟干旱的生理响应

研究了PEG模拟干旱胁迫环境下的火力楠(Michelia macclurel)、尾叶桉(Eucalyptus urophylla)、枫香(Liquidambar formosana)、荷木(Schima superba)幼苗的生理变化。结果表明,干旱胁迫下,4种幼苗叶片的相对含水量小于对照,其中,尾叶桉和枫香下降明显;不同干旱胁迫条件下,4种树种幼苗叶片的相对电导率均显著大于对照,其中尾叶桉和枫香上升幅度大;干旱胁迫下的火力楠和荷木幼苗叶片的脯氨酸含量呈现波动,尾叶桉和枫香幼苗则显著大于对照;不同干旱胁迫时间下的幼苗叶片的叶绿素含量小幅波动;4个树种幼苗的过氧岐化酶(SOD)活性随胁迫时间增加而呈现先升后降的趋势,其中火力楠和荷木的幼苗的SOD活性持续维持在较高水平;荷木叶片的丙二醛(MDA)含量先升后降,最后和对照水平相近,其余幼苗的MDA含量均大于对照;干旱胁迫下4种幼苗叶片的可溶性糖含量增加幅度较大。主成分分析表明,4种幼苗的抗旱能力排序为荷木>火力楠>尾叶桉>枫香。     

Characterization of protein release from hydrolytically degradable poly(ethylene glycol) hydrogels

We present a novel fully hydrophilic, hydrolytically degradable poly(ethylene glycol) (PEG) hydrogel suitable for soft tissue engineering and delivery of protein drugs. The gels were designed to overcome drawbacks associated with current PEG hydrogels (i.e., reaction mechanisms or degradation products that compromise protein stability): the highly selective and mild cross‐linking reaction allowed for encapsulating proteins prior to gelation without altering their secondary structure as shown by circular dichroism experiments. Further, hydrogel degradation and structure, represented by mesh size, were correlated to protein release. It was determined that polymer density had the most profound effect on protein diffusivity, followed by the polymer molecular weight, and finally by the specific chemical structure of the cross‐linker. By examining the diffusion of several model proteins, we confirmed that the protein diffusivity was dependent on protein size as smaller proteins (e.g., lysozyme) diffused faster than larger proteins (e.g., Ig). Furthermore, we demonstrated that the protein physical state was preserved upon encapsulation and subsequent release from the PEG hydrogels and contained negligible aggregation or protein–polymer adducts. These initial studies indicate that the developed PEG hydrogels are suitable for release of stable proteins in drug delivery and tissue engineering applications. Biotechnol. Bioeng. 2011; 108:197–206. © 2010 Wiley Periodicals, Inc.     

PEG引发对芹菜种子活力影响的研究

以芹菜种子为试验材料,研究了不同浓度的聚乙二醇（PEG）及其不同浸种时间对芹菜种子活力指标以及电导率的影响。结果表明,选择50mg／L的PEG处理芹菜种子能够显著提高其发芽率、发芽势、发芽指数、活力指数、芽长、根长、鲜重和干重。不同浓度PEG处理的芹菜种子的浸泡电导率和绝对电导率均显著低于对照组,且以50mg／L的PEG处理的两种电导率值最低。因此,通过选择50mg／LPEG浸泡芹菜种子4h,能够使其活力得到显著的提高。