基因芯片技术检测3种肠道病原微生物方法的建立

目的:建立一种运用多重PCR和基因芯片技术检测和鉴定伤寒沙门氏菌、痢疾杆菌和单核细胞增生利斯特菌的方法。方法:分别选取伤寒沙门氏菌染色体ViaB区域中编码调控Vi抗原表达的基因(vipR)、痢疾杆菌编码侵袭质粒抗原H基因(ipaH)和单核细胞增生利斯特菌溶血素基因(hlyA)设计引物和探针,探针3'端进行氨基修饰,下游引物标记荧光素Cy3。在优化的PCR和杂交反应条件下,进行三重PCR扩增,产物与包括3种致病菌特异性探针的基因芯片杂交。在评价基因芯片的特异性和灵敏度之后,对临床样本进行检测。结果:只有3种目的致病菌的PCR产物在相应探针位置出现特异性信号,其他阴性细菌均无信号出现;3种致病菌的检测灵敏度均可达到103CFU/mL;检测30例临床样本的结果与常规细菌学培养结果一致。结论:所建立的可同时检测伤寒沙门氏菌、痢疾杆菌和单核细胞增生利斯特菌的基因芯片方法快速、准确,特异性高,重复性好,为3种肠道致病菌的快速检测和鉴定提供了新方法和新思路。     

肺癌的表皮生长因子受体分子靶向治疗与基因突变

肺癌分子靶向治疗近年来取得较大进展,特别是针对表皮生长因子受体（EGFR）分子靶向药物表现出确定的临床效果。临床应用表明,EGFR基因酪氨酸激酶域体细胞突变与非小细胞肺癌患者对酪氨酸激酶抑制剂吉非替尼的敏感性相关,本文就相关的研究进行了简述。     

山罗花和天柱山罗花四个居群的数值分析

对山罗花和天柱山罗花的4个居群的15项形态特征进行了数值分析。结果表明:形态性状在4个居群间均存在一定程度的变异,居群间变异系数的平均值从小到大排列为:天柱山居群(天柱山罗花)、天柱山居群(山罗花)、黄山居群(山罗花)、鹞落坪居群(山罗花);天柱山罗花与山罗花种间的形态差异已达到极显著水平,山罗花的3个居群间部分性状也具有极显著的形态差异;以形态特征为基础的Q-聚类分析可以把天柱山罗花和山罗花聚为两类,但山罗花(天柱山居群)和其他2个居群在较远处聚在一起,R-聚类分析发现了强正相关关系、弱正相关关系、弱负相关关系的性状。     

Preparation and biomedical application of a non-polymer coated superparamagnetic nanoparticle

We report the preparation of a non-polymer coated superparamagnetic nanoparticle that is stable and biocompatible both in vitro and in vivo. The non-polymer, betaine, is a natural methylating agent in mammalian liver with active surface property. Upon systemic administration, the nanoparticle has preferential biodistribution in mammalian liver and exhibits good reduction of relaxivity time and negative enhancement for the detection of hepatoma nodules in rats using MRI. Our data demonstrate that the non-polymer coated superparamagnetic nanoparticle should have potential applications in biomedicine.     

Identification and functional analysis of LsMNPV anti-apoptosis genes

Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD(74) downward arrow G) that is different from Sl-P49 (TVID(94) downward arrow G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assay revealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.     

Function and oligomerization study of the leucine zipper-like domain in P13 from Leucania separata multiple nuclear polyhedrosis virus

The p13 gene is uniquely present in Group II nucleopolyhedroviruses (NPVs) and some granuloviruses, but not in Group I NPVs. p13 gene was first described by our laboratory in Leucania separatamultiple nuclear polyhedrosis virus (Ls-p13) in 1995. However, the functions of Ls-P13 and of its homologues are unknown. When Ls-p13 was inserted into Autographa californica nucleopolyhedrovirus, a Group I NPV, polyhedra yield was inhibited. However, this inhibition was prevented when the leucine zipper-like domain of Ls-p13 was mutated. To determine the cause of this marked difference between Ls-P13 and leucine zipper mutated Ls-P13 (Ls-P13mL), oligomerization and secondary structure analyses were performed. High performance liquid chromatography and yeast two-hybrid assays indicated that neither Ls-P13 nor Ls-P13mL could form oligomers. Informatics and circular dichroism spectropolarimetry results further indicated marked secondary structural differences between Ls-P13 and Ls-P13mL. The LZLD of Ls-P13 has two extended heptad repeat units which form a hydrophobic surface, but it is short of a third hydrophobic heptad repeat unit for oligomerization. However, the mutated LZLD of Ls-P13mL lacks the above hydrophobic surface, and its secondary structure is markedly different. This difference in its secondary structure may explain why Ls-P13mL is unable to inhibit polyhedra yield.     

Deferoxamine synergistically enhances iron-mediated AP-1 activation: a showcase of the interplay between extracellular-signal-regulated kinase and tyrosine phosphatase

Deferoxamine (DFO) is a drug widely used for iron overload treatment to reduce body iron burden. In the present study, it was shown in mouse epidermal JB6 cells that all iron compounds transiently induced extracellular signal-regulated kinases (ERK) phosphorylation, whereas DFO further enhanced ERK phosphorylation over long periods. The ERK phosphorylation by DFO treatment appears to be due to the inhibition of MAPK phosphatases (MKP) by DFO. The combined effects of iron-initiated MAPK activation and DFO-mediated MKP inhibition resulted in a synergistic enhancement on AP-1 activities. The results indicate that the interplay between MAPK and MKP is important in regulating the extent of AP-1 activation. It is known that administration of DFO in iron overload patients often results in allergic responses at the injection sites. The results suggest that this synergistic AP-1 activation might play a role in DFO-induced skin immune responses of iron overload patients.     

Role of Drosophila gene dunc-115 in nervous system

Axonal guidance signals are transduced through growth cone surface receptors to the interior leading to changes of actin dynamics and actin binding proteins, which are critical in determining the outcome of actin cytoskeleton reorganization. We report here the characterization of the Drosophila actin binding protein abLIM/Unc-115 homolog Dunc-115 and its role in the nervous system. Three Dunc-115 isoforms are identified as Dunc-115L, M and S, respectively. While Dunc-115L is a canonical homolog of Unc-115 with four LIM domains and one villin headpiece domain, Dunc-115M and S are novel isoforms without counterparts in other species. Our molecular modeling shows Dunc-115L is likely to bind to actin. Mutant analysis reveals that Dunc-115 is involved in axonal projection in both the visual and central nervous system.     

Intermedin/adrenomedullin 2及其受体在慢性低氧性肺动脉高压大鼠右心室的表达变化

本研究探讨了新发现的小分子生物活性肽intermedin／adrenomedullin 2（IMD／ADM2）及其受体在慢性低氧性肺动脉高压大鼠右心室中的变化和可能作用。用放射免疫分析法测定正常对照组和常压低氧4周组Sprague-Dawley大鼠血浆、右心室匀浆IMD／ADM2和肾上腺髓质素（adrenomednllin,ADM）蛋白水平;逆转录-多聚酶链反应法测定右心室IMD／ADM2、ADM及受体：降钙素受体样受体（calcitonin receptor-like receptor,CRLR）、受体活性修饰蛋白1,2,3（receptor activity modifying protein 1,2,3,RAMP1,RAMP2,RAMP3）mRNA表达。结果显示：低氧组平均肺动脉压、右心室与左心室加室间隔重量比[RV／（LV＋S）]显著高于对照组（均P〈0．01）;低氧组血浆和右心室组织匀浆ADM水平比对照组分别高1．26倍和1．68倍（P〈0．01）,IMD／ADM2水平则较对照组分别高0．90倍和1．19倍（P〈0．01）;与对照组相比,低氧组右心室IMD／ADM2、ADM mRNA表达均上调（P〈0．01）,RAMP2 mRNA表达增强（P〈0．05）,而两组间CRLR、RAMP1、RAMP3 mRNA的表达水平无显著性差异。结果表明,慢性低氧性肺动脉高压大鼠IMD／ADM2表达水平升高。