FS2M01．pl，转换片段大小表为0／0矩阵的Perl脚本

用Perl语言编写了一个脚本——FS2M01．pl来实现片段大小表向0／1矩阵的转换,解决了由人工方法将遗传图谱的位点信息转换成0／1矩阵的问题。     

小麦易位系1BL/1RS×7DL.7Ag的F2分子检测及其农艺和品质性状分析

以‘豫农982’(1BL/1RS易位)和wheatear(7DL.7Ag易位)杂交后代的900个F2群体及其F2∶3家系为实验材料,对F2群体进行1BL/1RS易位和7DL.7Ag易位类型分子检测,调查分析F2群体及F2∶3家系的农艺、产量性状(F2群体的农艺性状仅作参考,重点分析F2∶3家系的性状),并探讨1BL/1RS易位和7DL.7Ag易位对小麦品质的影响。结果表明:(1)STS标记Lr19130与SSR引物Xgwm428联合使用可作为共显性标记鉴定纯合7DL.7Ag易位与杂合7DL.7Ag易位,完善了7DL.7Ag易位的分子检测方法。(2)在农艺和产量性状方面,1BL/1RS易位可显著降低株高,有助于提高穗粒数和小穗数;7DL.7Ag易位在籽粒千粒重和产量上有显著的正向作用,但7DL.7Ag易位的穗粒数显著低于非7DL.7Ag易位且有延迟小麦成熟和增加株高的趋势;1BL/1RS和7DL.7Ag双重易位可同时提升小穗数和千粒重,但穗粒数减少。(3)在品质性状方面,1BL/1RS易位主要影响沉淀值、稳定时间、弱化度、最大拉伸阻力、延伸度等主要反映蛋白质质量的性状,使面筋质量显著降低,而对蛋白质含量、湿面筋含量和吸水率等主要反映蛋白质数量的性状影响较小;7DL.7Ag易位显著提高沉淀值和面团拉伸品质参数,对小麦粉质参数和糊化参数贡献不大。由此推测,将7DL.7Ag易位应用于小麦品种选育,有望突破产量瓶颈并可较好地提升品质。     

Mg2＋对阿霉素引起心肌线粒体F1F0变化的保护

抗肿瘤药物阿霉素(ADM)对心肌线粒体F₁F₀-复合体呈现抑制而对F₁-ATPase无抑制,这表明ADM可能是通过膜脂起作用的,适当浓度Mg²⁺能降低ADM对复合体的抑制.经 ³¹P-NMR和标记荧光探针NBD-PE,DPH,MC-540以及内源荧光等的测定,结果表明ADM可能首先通过诱导F₁F₀膜脂形成非双层脂结构,继而影响了膜脂的堆积程度和流动性,进而引起F₁F₀-复合体酶蛋白构象的改变,最终导致酶活力的降低.Mg^2＋则可能由于与ADM竞争与心磷脂的结合,而对ADM引起F₁F₀的变化产生保护作用.     

RNA干扰Chk1/2调控二烯丙基二硫诱导的G2/M期阻滞作用及相关周期蛋白表达

在细胞周期检测点信号传导通路中,Chkl和Chk2起着重要作用,主要参与G2/M期细胞周期检测点信号传导.首先采用RNAi技术在BGC823细胞中将Chk1、Chk2基因沉默,Chk1、Chk2 siRNA转染BGC823细胞后24h加入15mg/L二烯丙基二硫(diallyl disulfide,DADS),接着通过Real-timePCR、Western blot分析Chk1、Chk2基因在转染前后的表达差异,然后运用流式细胞术和Western blot分析Chk1、Chk2基因沉默后对DADS诱导的细胞周期G2/M期阻滞作用及相关周期蛋白CDC25C和cyclinB1表达的影响.实验结果表明,与未转染对照组相比,转染Chk1、Chk2siRNA后BGC823细胞中Chk1、Chk2表达明显被抑制,二者的mRNA表达分别下降84.7%和69.0%,蛋白质水平分别下降73.4%和78.5%.流式细胞术分析结果发现,ChklsiRNA转染的BGC823细胞在15mg/LDADS处理24h后,G2/M期比例由单纯DADS处理组的58.1%降至10.4%(P<0.05).但Chk2 siRNA转染后加入15mg/...     

0期诊断技术对军事训练所致椎间盘源性下腰痛患者血清IL-1β, 6-keto-PGF1α及TNF-α水平的影响及其意义

目的:探讨0期诊断技术对军事训练所致椎间盘源性下腰痛患者血清白介素1β(IL-1β)、6酮前列腺素F1α(6-keto-PGF1α)、肿瘤坏死因子α(TNF-α)水平的影响及其意义。方法:随机抽取某部2014年度新兵一营306名新兵为实验组,新兵二营290名新兵为对照组,在入伍复检时为每名士兵抽血检测相关血清学指标,并建立健康档案。由经过技术培训的医师对实验组采用0期诊断技术,分别在新兵训练开始后的第2、4、6、8、10、12周进行0期椎间盘源性下腰痛的筛查,并实施相应的防治措施,同时,按照"军标"对所有实验对象进行下腰痛的常规诊治及登统计,对于诊断为0期椎间盘源性下腰痛阳性和椎间盘源性下腰痛阳性的士兵及时抽血检测IL-1β、6-keto-PGF1α、TNF-α血清值。结果:实验组椎间盘源性下腰痛总发生率(2.94%)明显低于对照组(9.66%)(x~2=11.527,P0.001)。在新训开始后的第8、10、12周,实验组椎间盘源性下腰痛的发生率均明显低于对照组(P0.05);实验组和对照组阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α水平比较无差异,0期阳性和阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α水平与训练前相比均明显升高(P0.05),0期阳性士兵血清IL-1β、6-keto-PGF1α、TNF-α值均明显低于本组和对照组阳性士兵(P0.05)。结论:0期诊断技术预防军事训练所致新兵椎间盘源性下腰痛的效果明显,通过监测IL-1β、6-keto-PGF1α、TNF-α血清的水平变化也进一步证实0期诊断技术用于预防军事训练所致椎间盘源性下腰痛的可行性。     

巨噬细胞免疫调变信号——Raf-1,MAPK,cPLA2的激活和IL-12分泌的研究

LPS介导的小鼠腹腔巨噬细胞免疫调变的信号和机理是不清楚的.应用LPS和PMA处理抑制性巨噬细胞后,发现Ras下游信号分子Raf-1,MAPK和cPLA₂均被活化,包括Raf-1的磷酸化,MAPK p44和p42磷酸化以及cPLA_2的活化,使花生四烯酸的释放显著增加,结合新近发现的LPS、PMA能诱导抑制性巨噬细胞PKC-α和PKC-ε同工酶的激活与转位,认为在LPS介导的免疫调变中,PKC信号通路的活化及其与Raf-1/MAPK通路的“连接”是主要信号传导通路.同时调变巨噬细胞也能分泌IL-12.     

应用ELISPOT方法筛选确定HIV-1B'/C亚型疫苗六种抗原的H-2d限制的T细胞表位

为了筛选和确定用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、polr、evt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位,本研究使用表达上述6种抗原的复制型DNA疫苗和非复制型重组痘苗病毒疫苗联合免疫BALB/c小鼠,通过矩阵设计将HIV-1 B(C)亚型6种相应抗原全序列肽库分别混合成肽池,使用肽池对免疫小鼠进行IFN-γELISPOT检测,根据检测结果确定肽库中特异反应的优势表位肽。结果显示:筛选到七条针对Gag的特异表位肽,其中有5条与文献报道相同,另2条为新表位肽;筛选到3条针对Pol蛋白特异表位肽,其中一条为新表位肽;筛选到2条针对gp160特异表位肽,其中一条为新表位肽;在Nef肽库中筛选到一条新的表位肽;从Tat肽库中筛选到3条表位肽,这三条肽在肽库中是连续的序列,都包含(或部分包含)网上公布的表位序列;在Rev肽库中没有筛选到能够产生阳性反应的特异性表位肽。本研究使用IFN-γELISPOT方法筛选和确定了可用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、pol、revt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位。     

ChooseMaterials.pl，控制变量挑选实验材料的perl脚本

用Perl语言编写了一个脚本--ChooseMaterials．pl,通过计算两个材料特定某几列数据之间增加或减少的百分数水平,达到控制变量突出关键因子的作用,方便生物学工作者选取材料,做进一步的实验或分析。     

RBP-J is required for M2 macrophage polarization in response to chitin and mediates expression of a subset of M2 genes

Development of alternatively activated (M2) macrophage phenotypes is a complex process that is coordinately regulated by a plethora of pathways and factors. Here, we report that RBP-J, a DNA-binding protein that integrates signals from multiple pathways including the Notch pathway, is critically involved in polarization of M2 macrophages. Mice deficient in RBP-J in the myeloid compartment exhibited impaired M2 phenotypes in vivo in a chitin-induced model of M2 polarization. Consistent with the in vivo findings, M2 polarization was partially compromised in vitro in Rbpj-deficient macrophages as demonstrated by reduced expression of a subset of M2 effector molecules including arginase 1. Functionally, myeloid Rbpj deficiency impaired M2 effector functions including recruitment of eosinophils and suppression of T cell proliferation. Collectively, we have identified RBPJ as an essential regulator of differentiation and function of alternatively activated macrophages.     

Selection of peptide inhibitor to matrix metalloproteinase-2 using phage display and its effects on pancreatic cancer cell lines PANC-1 and CFPAC-1

Despite tremendous advances in cancer treatment and survival rates, pancreatic cancer remains one of the most deadly afflictions and the fourth leading cause of cancer deaths in the world. Matrix Metalloproteinases (MMPs) are thought to be involved in cancer progression. Matrix metalloproteinase (MMP)-2 is known to play a pivotal role in tumor invasion, metastasis and angiogenesis, and validated to be the anticancer target. Inhibition of MMP-2 activity is able to reduce the cancer cell invasion and suppress tumor growth in vivo. Two novel peptides, M204C4 and M205C4, which could specially inhibit MMP-2 activity, were identified by a phage display library screening. We showed that M204C4 and M205C4 inhibited the activity of MMP-2 in a dose dependent manner in vitro. Two peptides reduced MMP-2 mediated invasion of the pancreatic cancer cell lines PANC-1 and CFPAC-1, but not affected the expression and release of MMP-2. Furthermore, these two peptides could suppress tumor growth in vivo. Our results indicated that two peptides selected by phase display technology may be used as anticancer drugs in the future.     

与细胞周期G2/M期进程相关的H2AX磷酸化

γH2AX焦点(foci)被普遍当做DNA双链断裂（DSB）损伤的分子标志物.为探 讨细胞周期进程相关的H2AX磷酸化规律特征,采用胸腺嘧啶双阻滞结合噻氨酯哒唑(nocodazole)的后续处理,将HeLa细胞同步于有丝分裂的前中期．然后,用流式细胞仪检测细胞周期、Western印迹和免疫荧光法,观察γH2AX表达和γH2AX焦点的形成．结果显示,细胞进入G2/M期和有丝分裂过程中,γH2AX水平显著增加 ;在无DNA DSB发生的情况下,部分M期细胞中也存在大量的γH2AX焦点．随着细 胞完成有丝分裂从M期退出再进入G1期,γH2AX的表达水平逐渐降低．这种 γH2AX表达变化特征与G2/M期密切关联的PLK1和Cyclin B1的表达规律相类似． 在4 Gy大剂量照射下,HeLa细胞于照后8 到12 h出现明显的G2/M期阻滞．γH2AX 焦点数在照后1 h达高峰,随后降低,照后8 h又上升,出现了第2个峰值．与之不同的是,在1 Gy低剂量照射下,细胞的G2/M期阻滞微弱,γH2AX焦点数在照后 0.5 h最高,随后下降,且无反弹,符合DNA DSB的修复动力学特征．因此,将γ H2AX当做DNA DSB分子标志物时,还需要考虑细胞周期变化的影响．γH2AX适合 作为1 Gy以下照射的DNA双链断裂损伤的分子标志．     

The structural basis for high affinity binding of α1-acid glycoprotein to the potent antitumor compound UCN-01

The α1-acid glycoprotein (AGP) is an abundant blood plasma protein with important immunomodulatory functions coupled to endogenous and exogenous ligand-binding properties. Its affinity for many drug-like structures, however, means AGP can have a significant effect on the pharmokinetics and pharmacodynamics of numerous small molecule therapeutics. Staurosporine, and its hydroxylated forms UCN-01 and UCN-02, are kinase inhibitors that have been investigated at length as antitumour compounds. Despite their potency, these compounds display poor pharmokinetics due to binding to both AGP variants, AGP1 and AGP2. The recent renewed interest in UCN-01 as a cytostatic protective agent prompted us to solve the structure of the AGP2–UCN-01 complex by X-ray crystallography, revealing for the first time the precise binding mode of UCN-01. The solution NMR suggests AGP2 undergoes a significant conformational change upon ligand binding, but also that it uses a common set of sidechains with which it captures key groups of UCN-01 and other small molecule ligands. We anticipate that this structure and the supporting NMR data will facilitate rational redesign of small molecules that could evade AGP and therefore improve tissue distribution.     

A型流感病毒H5N1的M2离子通道稳定细胞株的建立

A型流感病毒H5N1的M2离子通道(H5M2)基因经优化后由人工合成,适合于哺乳动物细胞中表达.通过酶切克隆于pcDNA4质粒,并在HEK293细胞中建立稳定细胞株.Western blotting和免疫荧光证实H5M2在稳定细胞中只有在四环素诱导下才能表达,并经膜片钳证实在HEK293细胞中表达的H5M2具有H 通道活性,为M2离子通道功能的研究和M2离子通道阻断剂筛选方法的建立提供了参考.     

Relevance of histone H1 kinase activity to the G2/M transition during the cell cycle ofDictyostelium discoideum

The implication of histone H1 kinase activity for the G2/M transition during the cell cycle was investigated usingDictyostelium discoideum Ax-2. Histone H1 kinase with its activity was purified from cell extracts by the use of p13^suc1 affinity gel. In the vegetative cell cycle, the activity of histone H1 kinase including Cdc2 kinase was found using synchronized Ax-2 cells to be highest just before the entry into mitosis. The activity also was markedly enhanced just prior to the M phase from which developing cells (possibly prespore cells) reinitiate their cell cycle at the mound-tipped aggregate stage. These results strongly suggest the importance of Cdc2 kinase activity in the G2 to M phase transition during the cell cycle, as the case for other eukaryotic cells.     

外源突变基因AP1M2转化毛白杨的研究

为探究毛白杨开花调控分子机制并获得不飞絮毛白杨株系,利用农杆菌介导法将显性负突变结构基因爿Pm挖转入毛白杨中,经PCR检测,共获得阳性转化植株7株。通过RT_qPCR检测转基因植株中外源基因AP1M2表达量,发现各株系间的表达水平差异显著,最高为内参的5．41倍,最低为1．89倍。对转基因毛白杨中内源刚尸,和其他开花关键基因的表达量进行检测,发现内源AP1的表达受到抑制,表达量明显下调;其他开花关键基LFY、AP3、PI、SEP3和FTI等的表达量也有不同程度的降低,而FT2表达量并没有明显变化。这些研究结果表明转4尸m扭毛白杨中API、LFY、AP3、PI、SEP3和FT2的表达受到明显抑制．对毛白杨花发育将有一定影响。     

Development of M2BPGi: a novel fibrosis serum glyco-biomarker for chronic hepatitis/cirrhosis diagnostics

Many proteins in the living body are glycoproteins, which present glycans linked on their surface. Glycan structures reflect the degree of cell differentiation or canceration and are cell specific. These characteristics are advantageous in the development of various disease biomarkers. Glycoprotein-based biomarkers (glyco-biomarkers) are developed by utilizing the specific changes in the glycan structure on a glycoprotein secreted from the diseased cells of interest. Therefore, quantification of the altered glycan structures is the key to developing a new glyco-biomarker. Glycoscience is a relatively new area of molecular science, and recent advancement of glycotechnologies is remarkable. In the author’s institute, new glycoscience technologies have been designed to be efficiently utilized for the development of new diagnostic agents. This paper introduces a strategy for glyco-biomarker development, which was successfully applied in the development of Wisteria floribunda agglutinin-positive Mac-2 binding protein M2BPGi, a liver fibrosis marker now commercially available for clinical use.