Rad17 phosphorylation is required for claspin recruitment and Chk1 activation in response to replication stress

The ATR-mediated checkpoint is not only critical for responding to genotoxic stress but also essential for cell proliferation. The RFC-related checkpoint protein Rad17, a phosphorylation substrate of ATR, is critical for ATR-mediated checkpoint signaling and cell survival. Here, we show that phosphorylation of Rad17 by ATR is important for genomic stability and restraint of S phase but is not essential for cell survival. The phosphomutant Rad17AA exhibits distinct defects in hydroxyurea- (HU) and ultraviolet- (UV) induced Chk1 activation, indicating that separate Rad17 functions are required differently in response to different types of replication interference. Although cells expressing Rad17AA can initiate Chk1 phosphorylation after HU treatment, they fail to sustain Chk1 phosphorylation after withdrawal of HU and are profoundly sensitive to HU. Importantly, we found that phosphorylated Rad17 interacts with Claspin and regulates its phosphorylation. These findings reveal a phosphorylation-dependent function of Rad17 in an ATR-Rad17-Claspin-Chk1-signaling cascade that responds to specific replication stress.     

分子文库展示技术

分子文库展示技术是一系列广泛应用于多肽、蛋白质及药物筛选和研究蛋白质间相互作用的有效的生物学技术。它将组合成的具有一定长度的随机序列寡核苷酸片段(或cDNA)克隆到特定表达载体中,使其表达产物(多肽片段或蛋白质结构域)以融合蛋白的形式展示在活的噬菌体或细胞表面。根据其蛋白质表达是否依赖于宿主表达系统,分为体内表达展示系统和无细胞展示系统(体外表达展示系统)。就其展示的部位不同又可分为噬菌体展示技术、细胞表面展示技术、核糖体展示技术、mRNA展示技术等。现对各种展示技术的基本原理及相关应用做简要综述。     

In vitro and in vivo effects of hydrolysates from conglycinin on intestinal microbial community of mice after Escherichia coli infection

AIMS: To detect the effect of pepsin-hydrolysate conglycinin (PTC) on the growth of Escherichia coli O(138)in vitro, and investigate the effect of PTC on intestinal microbial community of mice after E. coli infection. METHODS AND RESULTS: Serial dilution method was used to detect the antibacterial activity of PTC in 96-well cell-cultivated plates. Fifty-five KM mice were randomly assigned to five groups: normal, feeding-E. coli control, HCl-full hydrolysis of conglycinin, conglycinin and PTC. Orally administrated with hydrolysates from conglycinin for 21 days, each mouse was fed with 2 x 10(8) CFU ml(-1) of E. coli O(138) on the 22nd day. The mice activities were monitored and polymerase chain reaction-denaturing gradient gel electrophoresis was used to analyse the microbial community in mice faeces. The results showed that PTC could inhibit growth of E. coli O(138) at nitrogen concentrations of more than 520 mg l(-1). There was high similarity of intestinal microbial community in mice between PTC and normal groups. CONCLUSION: PTC inhibits growth of E. coli O(138), keeps mice healthy following oral administration of E. coli infection and maintains a balanced active microbial community in their gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the antibacterial activity of PTC against E. coli and its ability to maintain healthy intestinal microbial community in mice even after they were infected with E. coli. This observation is significant in the application of PTC to prevent gastrointestinal diseases caused by E. coli and unbalanced intestinal microflora.     

IL-1β在缺血致脑损伤中的主要作用机制

中枢白细胞介素-1β（IL-1β）在缺血引起的脑损伤过程中发挥十分重要的作用,但其作用机制尚不十分明确,本文主要就近年来的研究进展,从其对诱导粘附分子表达,激活内皮细胞产生多种活性物质,促进脑内兴奋性氨基酸和自由基等神经毒性物质的产生和释放等角度,简要概括其在缺血引起的脑损伤过程中的作用机制。     

Discovery of 5-(arenethynyl) hetero-monocyclic derivatives as potent inhibitors of BCR-ABL including the T315I gatekeeper mutant

Ponatinib (AP24534) was previously identified as a pan-BCR-ABL inhibitor that potently inhibits the T315I gatekeeper mutant, and has advanced into clinical development for the treatment of refractory or resistant CML. In this study, we explored a novel series of five and six membered monocycles as alternate hinge-binding templates to replace the 6,5-fused imidazopyridazine core of ponatinib. Like ponatinib, these monocycles are tethered to pendant toluanilides via an ethynyl linker. Several compounds in this series displayed excellent in vitro potency against both native BCR-ABL and the T315I mutant. Notably, a subset of inhibitors exhibited desirable PK and were orally active in a mouse model of T315I-driven CML.     

短序与阔叶十大功劳化学组分及小檗碱含量的FTIR比较研究

为快速评价短序与阔叶十大功劳不同部位化学成分差异,本研究运用FTIR采集短序和阔叶十大功劳不同部位的红外光谱并比较短序与阔叶十大功劳不同部位盐酸小檗碱含量差异.研究结果显示,生物碱、黄酮和丁香酯等化学成分,短序十大功劳根、茎及叶片中均较阔叶十大功劳丰富,但是,多糖和苷类含量在两种十大功劳根中相当,在茎和叶片中均以阔叶十大功劳含量高,尤其是叶片中这种差异更明显.最后,与盐酸小檗碱标准品比较发现,2种十大功劳中盐酸小檗碱含量均以茎中最高,叶片中含量最低;2种十大功劳比较各部位盐酸小檗碱含量均以短序十大功劳中高.故运用FTIR技术可以快速找出2种十大功劳化学成分的差异,本研究结果将为十大功劳属植物资源合理开发利用及良种选育提供参考.     

微生物菌种改良的新方法新策略

自然分离得到的原始菌种远不能达到工业生产要求,通过菌种改良获得高产菌种是有效的手段。传统方法虽然无需了解过多遗传背景就能取得成效,但往往耗时费力。随着DNA重组技术、原生质体融合、组学研究的应用日益广泛,微生物菌种改良的新方法和新策略诸如代谢工程、基因组改组和系统生物技术、核糖体工程、表观遗传修饰等逐步发展起来。以下综述了近年来菌种改良相关领域方法和策略特别是首次报道的表观遗传修饰的最新进展。     

Experimental study and mechanism analysis on bioeffects by nanosecond electromagnetic pulses

The athermal bioeffects caused by nanosecond electromagnetic pulses with body cells was studied by using a broad band transverse EM-wave cell (BTEM CELL). The experimental system and preliminary mechanism analysis were presented.     

准分子激光治疗近视眼

本文对准分子激光PRK治疗近视眼息者503例的早期临床结果进行分析,近视范围为－2．0至－15．0D,根据术前屈光度不同分为三组,术后随访三个月．结果证实准分子激光是一种安全、有效、预测性较好的治疗近视眼方法．全部病人提高了裸眼视力,绝大部分保持了最佳矫正视力,且没有产生危及视力的并发症．