Characterization of the 3a protein of SARS-associated coronavirus in infected vero E6 cells and SARS patients

Proteomics was used to identify a protein encoded by ORF 3a in a SARS-associated coronavirus (SARS-CoV). Immuno-blotting revealed that interchain disulfide bonds might be formed between this protein and the spike protein. ELISA indicated that sera from SARS patients have significant positive reactions with synthesized peptides derived from the 3a protein. These results are concordant with that of a spike protein-derived peptide. A tendency exists for co-mutation between the 3a protein and the spike protein of SARS-CoV isolates, suggesting that the function of the 3a protein correlates with the spike protein. Taken together, the 3a protein might be tightly correlated to the spike protein in the SARS-CoV functions. The 3a protein may serve as a new clinical marker or drug target for SARS treatment.     

重引入狗獾秋冬季行为的初步研究

2010年11月至2011年2月在上海奉贤申亚生态园内,对从山东重引入的狗獾(Meles meles)在围栏内进行了秋冬季行为的初步研究。结果表明,重引入狗獾在冬季活动时间低,取食食物以肉类饲料为主,平均2～4 d出洞活动一次,受温度影响大。研究表明,重引入狗獾能适应人工饲养并顺利越冬。     

Chromosome dynamic changes in two cultured chinese human embryonic stem cell lines: Single nucleotide polymorphism, copy number variation and loss of heterozygosity

The quality and safety of human embryonic stem cells (hESCs) in clinical application depend on gene stability. Two Chinese hESC lines, Zh1 and Zh21, were incubated over a long period. We observed and compared the gene stability in the passage numbers 20, 17 for Zh1 cell line and passage numbers 27, 60, 68 for Zh21 cell line. Single nucleotide polymorphisis analysis indicated that hESCs in early passages had relative gene stability; and with the increase in passage number, gene instability became strong. We also found that there were copy number variations (CNVs) in both Zh21 and Zh1. We analyzed the CNVs of Chinese Han Beijing man (CHB; normal Chinese people) and found that the all CNV forms were the loss in Zh21, Zh1, and CHB. We also analyzed and compared the related pathways of the mutant genes. We propose three steps to ensure hESC safety. Firstly, besides the conventional methods such as pluripotent genes, chromosome G‐banding and teratoma, high‐resolution DNA chip analysis should also be adopted; secondly, chromosomal properties are monitored every 10 passages in less than passage 50 and every 5 passages in more than passage 50; thirdly, the related pathways of mutant genes should be observed because only the mutant genes with variations of their related pathways may affected cell functions. J. Cell. Biochem. 113: 3520–3527, 2012. © 2012 Wiley Periodicals, Inc.     

Novel stimuli-responsive micelle self-assembled from Y-shaped P(UA-Y-NIPAAm) copolymer for drug delivery

A new amphiphilic Y-shaped copolymer, comprised of hydrophobic poly(undecylenic acid) (PUA) and hydrophilic poly(N-isopropylacrylamide) (PNIPAAm), was designed and synthesized. A cytotoxicity study revealed that P(UA-Y-NIPAAm) copolymers did not exhibit apparent inhibition impact on the proliferation of cells when the concentration of the copolymer was below 1000 mg/L. Characterization demonstrated that the P(UA-Y-NIPAAm) copolymer is thermosensitive with a lower critical solution temperature (LCST) of 31 degrees C. In water, the P(UA-Y-NIPAAm) copolymer would self-assemble into micelles with a critical micelle concentration (CMC) of 20 mg/L. Self-assembled P(UA-Y-NIPAAm) micelles exhibited a nanospherical morphology of 40 to approximately 80 nm in size. The controlled drug release behavior of the P(UA-Y-NIPAAm) micelles was further investigated, and self-assembled micelles exhibited improved properties in controlled drug release.     

p53与Ras协同及其在肿瘤发生中的作用

肿瘤发生是抑癌基因失活和原癌基因激活共同作用的结果。p53基因被认为是目前最重要的抑癌基因,50%以上的肿瘤中存在p53基因的点突变现象;而Ras基因是肿瘤中突变率较高的原癌基因,其突变率在某些肿瘤中高达30%-90%。研究发现,肿瘤发生过程中抑癌基因p53与原癌基因Ras之间存在复杂的相互协同作用。根据目前的文献报道,p53与Ras之间的协同作用可以分为3种:第一,p53对Ras的调节作用;第二,Ras对p53的调节作用;第三,p53和Ras共同调控某些与肿瘤发生相关的关键基因。了解p53与Ras之间的3种调控作用将有助于我们进一步认识p53失活与Ras激活协同促进肿瘤发生的分子通路和机制,同时也将为癌症的个性化治疗和药物靶点的选择提供重要依据。因此,文章将对近年来所发现的p53与Ras的各种协同作用机制及其与肿瘤发生的关系进行概括和综述。     

Synthesis,DNA-binding abilities and anticancer activities of triazole-pyrrolo[2,1-c][1,4]benzodiazepines hybrid scaffolds

We synthesized a new series of PBD-hybrid derivatives having tethered triazoles and investigated for their cytotoxicity. The studies indicated that cis-olefin compounds induce higher cytotoxicity with increase in the G1 cell cycle phase compared with the trans-compounds. Quantitative RT-PCR assay indicated that compounds (16a–d) induced G1 phase arrest through down-regulation of cyclin D1 and up-regulation of p21, p27, and p53 mRNA expressions. Compounds 16a–d induced A375 early apoptosis as detected by flow cytometry after double-staining with annexin V and propidium iodide. Moreover, the Western blot analysis showed that A375 treated by compounds (16a–d) resulted in decreased levels of Bcl-2 and Bcl-xL, increased levels of Bax and Bad, and caspase/PARP degradation to identify apoptotic cells.     

烯啶虫胺肠衣饵剂对黑翅土白蚁的防治效果

[目的]为了开发一种高效环保型白蚁饵剂.[方法]选定烯啶虫胺和溴虫腈对黑翅土白蚁Odontotermes formosanus进行室内毒力测定,根据室内毒力测定结果筛选毒力较高药剂进行驱避试验,并制备3种不同剂量的肠衣饵剂,开展室内白蚁毒杀效果试验和园林白蚁防治效果试验;选用合适剂量肠衣饵剂进行堤坝黑翅土白蚁诱杀试验,并用挖巢法检测肠衣饵剂对堤坝黑翅土白蚁的诱杀效果.[结果]1.6tg/mL烯啶虫胺处理黑翅土白蚁72 h后校正死亡率达100％,16 μtg/mL溴虫腈处理72 h后黑翅土白蚁校正死亡率也达到100％.室内毒力测定结果表明烯啶虫胺对黑翅土白蚁的LC5o值低于溴虫腈,100 μtg/mL烯啶虫胺处理8h后对黑翅土白蚁无显著驱避作用.3种剂量烯啶虫胺肠衣饵剂处理72 h后,黑翅土白蚁校正死亡率均在60％以上.将不同剂量的烯啶虫胺肠衣饵剂投放到园林中45 d后,肠衣饵剂基本被食空,施药点周围无白蚁及活动迹象.在福建省水库大坝周围投放60 μtg/g烯啶虫胺肠衣饵剂,3个月后肠衣饵剂未发霉且基本被食空,6个月后挖巢发现蚁道内无黑翅土白蚁活动,白蚁巢体出现死亡和坍塌情况.[结论]60 μg/g烯啶虫胺肠衣饵剂对园林和堤坝中的黑翅土白蚁具有良好的诱杀效果,是一种高效环保型白蚁肠衣饵剂.     

Enantioselective hydrolysis of naproxen ethyl ester catalyzed by monoclonal antibodies

This report described that a hapten of racemic phosphonate 3 designed as the mimic of the transition state of hydrolysis of naproxen ethyl ester was successfully synthesized from easily available 2-acetyl-6-methoxy-naphthalene 5. Then BALB/C mice were immunized and one of the monoclonal catalytic antibodies, N116-27, which enantioselectively accelerated the hydrolysis of the R-(-)-naproxen ethyl ester was given. The Michaelis-Menton parameter for the catalyzed reaction was K(M)=6.67 mM and k(cat)/k(uncat)=5.8 x 10(4). This enantioselective result was explained by the fact that the R-isomer of rac-hapten was more immunogenic than the S-isomer.     

Upregulation of Nav1.3 Channel Induced by rrTNF in Cultured Adult Rat DRG Neurons via p38 MAPK and JNK Pathways

Activation of p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal protein kinase (JNK) in the dorsal root ganglia (DRG) is critical for the development of neuropathic pain. Tetraodontoxin-sensitive Nav1.3 channel, expressed at a very low level in the adult nervous system, is up-regulated in DRG neurons after peripheral nerve injury or peri-sciatic administration of rat recombinant tumour necrosis factor-alpha (rrTNF-α). To test if activation of p38 MAPK and JNK is required for the re-expression of Nav1.3 channel in cultured adult rat DRG neurons, we administrated rrTNF to cultured adult rat DRG neurons to induce Nav1.3 re-expression, and pre-treated with p38 MAPK inhibitor (SB203580 at 2.65, 26.5 and 265 μM) or JNK inhibitor (SP600125 at 1, 10 and 100 μM) 2 h before rrTNF to observe changes of Nav1.3-immunoreactivity. Compared with the DMSO vehicle pre-treatment group, SB203580 at 2.65 μM partially blocked the re-expression of Nav1.3 (P<0.001), and at 26.5 and 265 μM completely blocked Nav1.3 (P<0.001). Similarly, SP600125 at the concentration of 1 μM blocked the re-expression of Nav1.3 partially (P<0.001), and at 10 and 100 μM blocked Nav1.3 completely (P<0.001). These data show that the activation of both p38 MAPK and JNK in DRG neurons was involved in the re-expression of Nav1.3 channel triggered by TNF-α, which might contribute to neuropathic pain.     

Basic Fibroblast Growth Factor Stimulates the Proliferation of Bone Marrow Mesenchymal Stem Cells in Giant Panda (Ailuropoda melanoleuca)

It has been widely known that the giant panda (Ailuropoda melanoleuca) is one of the most endangered species in the world. An optimized platform for maintaining the proliferation of giant panda mesenchymal stem cells (MSCs) is very necessary for current giant panda protection strategies. Basic fibroblast growth factor (bFGF), a member of the FGF family, is widely considered as a growth factor and differentiation inducer within the stem cell research field. However, the role of bFGF on promoting the proliferation of MSCs derived from giant panda bone marrow (BM) has not been reported. In this study, we aimed to investigate the role of bFGF on the proliferation of BM-MSCs derived from giant panda. MSCs were cultured for cell proliferation analysis at 24, 48 and 72 hrs following the addition of bFGF. With increasing concentrations of bFGF, cell numbers gradually increased. This was further demonstrated by performing 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) cell proliferation assay, 5-Bromo-2-deoxyUridine (BrdU) labeling and cell cycle testing. Furthermore, the percentage of MSCs that were OCT4 positive increased slightly following treatment with 5 ng/ml bFGF. Moreover, we demonstrated that the extracellular signal-regulated kinase (ERK) signaling pathway may play an important role in the proliferation of panda MSCs stimulated by bFGF. In conclusion, this study suggests that giant panda BM-MSCs have a high proliferative capacity with the addition of 5 ng/ml bFGF in vitro.