High level soluble expression and one-step purification of IBDV VP2 protein in <Emphasis Type="Italic">Escherichia coli</Emphasis>

Objectives

To improve the expression of soluble IBDV VP2 protein by using different tagged vectors in Escherichia coli.

Results

Fusion tags, Grifin, MBP, SUMO, thioredoxin, γ-crystallin, ArsC and PpiB, enhanced the expression and solubility of VP2 protein. The fusion proteins were purified by Ni–NTA chromatography, MBP-VP2 showed the highest purity about 90 %. After removing the MBP tag, VP2 self-assembled into virus-like particles, ~25 nm diam. Results from AGP suggested the recombinant IBDV VP2 protein identified by reference serum like IBDV.

Conclusion

All the seven tags enhanced the expression and solubility of IBDV VP2 protein. The recombinant protein self-assembly into virus like particles and possess antigenicity as reference IBDV.

     

Expression of chemokine-like factor 1 is upregulated during T lymphocyte activation

Chemokine-like factor 1 (CKLF1) is a cytokine with chemotactic effects on leukocytes and a functional ligand of CCR4. This cytokine is widely expressed and the level of expression is reported to be upregulated in asthma and rheumatoid arthritis (RA), disease conditions in which T lymphocytes are over-activated. In order to determine the expression profile of CKLF1 in activated T lymphocytes, we first employed a PCR-based method on human blood fractions cDNA panels and found that CKLF1 was upregulated in activated CD4+ and CD8+ cells, with no obvious changes in CD19+ cells. We further performed kinetic analyses of CKLF1 expression in phytohemagglutinin (PHA)-stimulated human peripheral blood lymphocytes (PBL) at both the mRNA and protein levels. In resting PBL, the constitutive expression of CKLF1 was low at mRNA level and barely detectable at the protein level; however, both were remarkably upregulated by PHA, appearing at 8h after PHA-stimulation and persisting up to 72h. These results suggest that CKLF1 may be involved in T lymphocyte activation and further study of CKLF1 function will prove valuable.     

探讨模拟冷空气降温过程对健康大鼠和高血压大鼠凝血功能的影响

目的:通过模拟冷空气温度变化过程给予健康大鼠和高血压大鼠冷刺激,以此探讨冷空气过程对机体凝血功能的影响。方法:收集张掖市2011年3月一次典型冷空气过程数据,利用气象环境模拟箱模拟其温度变化过程。将24只雄性健康大鼠和24只雄性高血压大鼠分别随机分成最低温组(Tmin组)、Tmin对照组、复温组(Tr组)和Tr对照组。将Tmin组和Tr组大鼠放入气候箱中暴露冷空气温度变化过程。在Tmin和Tr时点分别停止Tmin组大鼠和Tr组大鼠冷空气暴露,并采血以测定其凝血功能指标—凝血四项。结果:与对照组相比,健康大鼠和高血压大鼠活化部分凝血酶原时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)在降温后与对照组相比没有显著差异(P>0.05),但血浆纤维蛋白原(Fbg)含量在高血压大鼠和健康大鼠Tmin组明显高于其对照组(P<0.01)。温度恢复后,其在健康组含量仍然高于其对照组(P<0.05),而在高血压大鼠中没有差异(P>0.05)。同Fbg,反应Fbg的纤维蛋白原时间(Fbg-time)在健康大鼠Tmin和Tr组中短于对照组(P<0.01,),而在高血压大鼠中仅在Tmin组短于对照组(P<0.01)。高血压大鼠血中的Fbg含量和Fbgt明显高于和短于健康大鼠(P<0.01)。结论:①冷空气降温过程能增加机体血中Fbg含量,使凝血功能增强,可能增加心血管疾病危险性;②冷空气刺激对健康大鼠凝血功能影响强于高血压大鼠。     

Local habitat condition rather than geographic distance determines the genetic structure of Tamarix chinensis populations in Yellow River Delta,China

Tree Genetics &; Genomes - To breed a new variety of coffee (Coffea arabica) requires approximately 25&;nbsp;years due to the long generation time (5–6&;nbsp;years) of this perennial...     

沉默交配型信息调节因子2同源蛋白Sirt1

沉默交配型信息调节因子2同源蛋白1(silent mating type information regulation 2 homolog 1,Sirt1)是哺乳动物中与酵母沉默信息调节蛋白2(silencing information regulator 2,Sir2)高度同源的蛋白质,它是一种依赖NAD+的III类组蛋白去乙酰化酶(HDAC III),在细胞分化、衰老、凋亡、DNA损伤修复、能量及内分泌代谢调节中起重要作用,同时在基因沉默、表观遗传学修饰、转录调控及信号转导调节中发挥重要的生物学功能。本文对其近年的研究进展做一概述。     

中国棉铃虫核型多角体病毒DNA聚合酶基因的克隆和序列分析

应用谷实夜蛾核型多角体病毒（ＨｚＳＮＰＶ）ＤＮＡ聚合酶基因ＨｉｎｄⅢ／ＰｓｔⅠ３５９６ｂｐ片段作探针,经Ｓｏｕｒｔｈｅｒｎｂｌｏｔ杂交,克隆了中国棉铃虫核型多角体病毒（ＨａＳＮＰＶ）完整的ＤＮＡ聚合酶基因,大小约为３．４ｋｂ。限制性内切酶分析表明,ＨａＳＮＰＶＤＮＡ聚合酶基因限制性内切酶图谱与ＨｚＳＮＰＶ相似。用双脱氧链终止法测定该基因部分核苷酸序列（８０５ｂｐ）,推导出编码区２０６ａ。序列同源性比较显示,ＨａＳＮＰＶＤＮＡ聚合酶与ＨｚＳＮＰＶ之间具有高度的同源性;与ＬｄＭＮＰＶ、ＡｃＭＮＰＶ、ＢｍＳＮＰＶ、ＣｆＭＮＰＶ和ＯｐＭＮＰＶ也具有一定的同源性     

Discovery of new thienopyrimidine derivatives as potent and orally efficacious phosphoinositide 3-kinase inhibitors

A series of new thienopyrimidine derivatives has been discovered as potent PI3K inhibitors. The systematic SAR studies for these analogues are described. Among them, 8a and 9a exhibit nanomolar enzymatic potencies and sub-micromolar cellular anti-proliferative activities. 8a displays favorable pharmacokinetic profiles, while 9a easily undergoes deacetylation to yield a major metabolite 8a. Furthermore, 8a and 9a potently inhibit tumor growth in a dose-dependent manner in the NCI-H460 xenograft model with an acceptable safety profile.     

Iron metabolism in diabetes-induced Alzheimer’s disease: a focus on insulin resistance in the brain

Alzheimer’s disease (AD) is characterized by an excessive accumulation of toxic amyloid beta (Aβ) plaques and memory dysfunction. The onset of AD is influenced by age, genetic background, and impaired glucose metabolism in the brain. Several studies have demonstrated that diabetes involving insulin resistance and glucose tolerance could lead to AD, ultimately resulting in cognitive dysfunction. Even though the relationship between diabetes and AD was indicated by significant evidences, the critical mechanisms and metabolic alterations in diabetes induced AD are not clear until now. Recently, iron metabolism has been shown to play multiple roles in the central nervous system (CNS). Iron deficiency and overload are associated with neurodegenerative diseases. Iron binds to Aβ and subsequently regulates Aβ toxicity in the CNS. In addition, previous studies have shown that iron is involved in the aggravation of insulin resistance. Considering these effects of iron metabolism in CNS, we expect that iron metabolism may play crucial roles in diabetic AD brain. Thus, we review the recent evidence regarding the relationship between diabetes-induced AD and iron metabolism.