Analysis of Expressed Sequence Tags in Porcine Uterus Tissue

Two non-normalized cDNA libraries of uteri from Danish Landrace and Chinese Erhualian pigs were constructed, and 13,756 expressed sequence tags (ESTs) were randomly sequenced. The ESTs were clustered by Phrap software, and 6,139 distinct tentative consensus sequences were produced, including 2,730 contigs and 3,409 singlets. Using Blast tools, these 6,139 candidate genes were compared to the nr and nt databases; 5,210 of them were assigned putative functions, whereas 929 potentially represent new genes. Highly expressed genes appear to be associated with basic energy metabolism, transferase activity, localization, cellular physiological process, protein binding, and nucleic acid binding. Antileukoproteinase was the most highly expressed gene, corresponding to endometrial differentiation and conceptus or fetal development.     

Blockade of sphingosine-1-phosphate reduces macrophage influx and retinal and choroidal neovascularization

Sphingosine-1-phosphate (S1P) is a bioactive lipid molecule that stimulates endothelial cell migration, proliferation, and survival in vitro, and tumor angiogenesis in vivo. In this study, we used a humanized monoclonal antibody (sonepcizumab) that selectively binds S1P to investigate its role in retinal and choroidal neovascularization (NV). Intraocular injection of sonepcizumab significantly reduced macrophage influx into ischemic retina and strongly suppressed retinal NV in mice with oxygen-induced ischemic retinopathy. In mice with laser-induced rupture sites in Bruch's membrane, intraocular injection of sonepcizumab significantly reduced the area of choroidal NV and concomitantly reduced fluorescein leakage from the remaining choroidal NV. Four weeks after intraocular injection of up to 1.8 mg of the sonepcizumab in non-human primates, electroretinograms and fluorescein angiograms were normal, and light microscopy of ocular sections showed no evidence of structural damage. These data show for the first time that S1P stimulates both choroidal and retinal NV and suggest that sonepcizumab could be considered for evaluation in patients with choroidal or retinal NV.     

EB病毒潜伏膜蛋白LMP1编码基因沉默对NF—kB表达的影响

目的：探讨EBV阳性胃上皮细胞中潜伏膜蛋白1（latentmembraneprotein-1,LMP1）基因沉默对NF-kB转录表达的影响。方法：以稳定表达LMP1的EBV阳性胃上皮细胞系作为靶细胞,采用化学合成的siRNA特异性沉默LMP1,用RT-PCR和Westem-blotting分别检测其在不同时间段mRNA和蛋白水平特异性沉默效果;Westernblotting及免疫酶染色法检测LMP1基因沉默对NF—kB转录表达及核转移的影响。结果：siRNA在mRNA和蛋白水平可特异性沉默LMP1的表达,且有时间效应关系;Westem blotting及免疫酶染色法检测结果显示LMP1沉默可干扰NF—KB表达,导致靶细胞核内NF-kB水平下降,细胞浆内水平升高,而NF—kB总蛋白有下降趋势。结论：LMP1基因特异性沉默能够影响NF-kB表达,抑制其核转移。     

苹果盘二孢的分离培养研究

苹果盘二孢Marssonina coronaria引起的褐斑病是造成我国苹果树早期落叶的主要原因,由于该病菌分离培养困难,阻碍了对其生物学特性的研究,进而影响了对其防治机理和流行规律的研究。本研究应用4种培养基质,探索了3种方法对苹果盘二孢的分离效果。结果表明,3种方法均可分离到病原菌,但组织块分离法和分生孢子团分离法成功率仅有10%左右,而单孢分离法污染少,成功率高达到90%以上,明显优于其他两种方法。不同培养基上菌落形态、大小和产孢情况差异也很大,培养1个月(25℃)后PDA上菌落黑褐色隆起,表面蚯蚓粪状,无气生菌丝,无子实体和基内菌丝;10%V8培养基上菌落中央隆起,黑褐色,表面生少量气生菌丝,边缘放射状,基内菌丝深褐色,有子实体;苹果叶片葡萄糖琼脂培养基(LDA)上菌落平坦,黄褐色,表面生茂密的金黄色气生菌丝,基内菌丝深褐色,有子实体;苹果叶片煎汁葡萄糖琼脂培养基(LEDA)上菌落有明显的不规则隆起,黄褐色至黑褐色,表面生少许气生菌丝,菌落生长缓慢,无基内菌丝,分生孢子盘菌落表面生,菌落直径仅2mm左右,而在其他培养基上的菌落直径可达6-8mm,说明培养基质、分离方法均对苹果盘二孢的分离培养和生长发育有明显的影响。     

Doppel-induced cytotoxicity in human neuronal SH-SY5Y cells is antagonized by the prion protein

Doppel （Dpl） is a prion （PrP）-like protein due to the structural and biochemical similarities; however, the natural functions of Dpl and PrP remain unclear. In this study, a 531-bp human PRND gene sequence encoding Dpl protein was amplified from human peripheral blood leucocytes. Furl-length and various truncated human Dpl and PrP proteins were expressed and purified from Escherichia coil Supplement of the full-length Dpl onto human neuroblastoma cell SH-SY5Y induced remarkable cytotoxicity, and the region responsible for its cytotoxicity was mapped at the middle segment of Dpl [amino acids （aa） 81-122]. Interestingly, DpMnduced cytotoxicity was antagonized by the presence of full- length wild-type PrP. Analysis on fragments of PrP mutants showed that the N-terminal fragment （aa 23- 90） of PrP was responsible for the protective activity. A truncated PrP （PrPA32-121） with similar secondary structure as Dpl induced DpMike cytotoxicity on SH- SY5Y cells. Furthermore, binding of copper ion could enhance the antagonizing effect of PrP on Dpi-induced cytotoxicity. Apoptosis assays revealed that cytotoxicity induced by Dpl occurred through an apoptotic mechanism. These results suggested that the function of Dpl is antagonistic to PrP rather than synergistic.     

Eicosanoids mediate nodulation reactions to bacterial Escherichia coli K 12 infections in larvae of the oriental blowfly, Chrysomya megacephala

Abstract  Nodulation is the predominant cellular defense reaction to bacterial challenges in insects. In this study, third instar larvae of Chrysomya megacephala were injected with bacteria, Escherichia coli K 12 (10⁶ CFU/mL, 2 μL), immediately prior to injection of inhibitors of eicosanoid biosynthesis, which sharply reduced nodulation response. Test larvae were treated with specific inhibitors of phospholipase A₂ (dexamethasone), cyclo-oxygenase (indomethacin, ibuprofen and piroxicam), dual cyclo-oxygenase/lipoxygenase (phenidone) and lipoxygenase (esculetin) and these reduced nodulation except esculetin. The influence of bacteria was obvious within 2 h of injection (5 nodules/larva), and increased to a maximum after 8 h (with 15 nodules/larva), and then significantly reduced over 24 h (9 nodules/larva). The inhibitory influence of dexamethasone was apparent within 2 h of injection (4 vs. 5 nodules/larva), and nodulation was significantly reduced, compared to control, over 24 h (5 vs. 8 nodules/larva). Increased dosages of ibuprofen, indomethacin, piroxicam and phenidone led to decreased numbers of nodules. Nodules continued to exist during the pupal stage. However, the effects of dexamethasone were reversed by treating bacteria-injected insects with an eicosanoid-precursor polyunsaturated fatty acid, arachidonic acid. These findings approved our view that eicosanoid can mediate cellular defense mechanisms in response to bacterial infections in another Dipteran insect C. megacephala .     

中国黄耆属簇毛黄耆亚属的叶表皮特征及其系统学意义

在光学显微镜和扫描电镜下,观察了国产黄耆属簇毛黄耆亚属16种1变种的叶表皮特征。结果表明,气孔器在各种植物的上、下表皮均有分布,多为无规则型,也有不等细胞型;叶表皮细胞形状有不规则形与多边形,表皮细胞垂周壁有平直、浅波状、波状或深波状。叶表皮细胞形状与垂周壁的式样可以分为四种类型,这四种类型与亚属的分组有一定的对应关系。在扫描电镜下可见表皮细胞上有角质层,气孔下陷,气孔的外拱盖及其内缘特征在亚属内都比较一致。表皮细胞角质层的纹饰在个别类群中有一定的变异,对种类的鉴定有一定的意义。     

定量聚合酶链反应评估去白细胞滤器去除血液成分中巨细胞病毒的效果

为了评估目前常用的去白细胞滤器去除血液成分中人巨细胞病毒( HCMV) 的效果, 我们在献血员中筛选获得HCMV-IgG、pp65 皆阴性的血液, 分离外周血单个核细胞( PBMC) , 将HCMV 感染的人成纤维细胞与PBMC 共培养, 感染HCMV 的PBMC 经抗人成纤维抗体和抗CD45 磁珠2 次分选纯化后, 将CD45 + 单核细胞加回原全血中, 建立HCMV 潜伏感染的模型, 再用去白细胞滤器过滤。用实时聚合酶链反应( PCR) 定量测定过滤前、后血液样本中的病毒载量, 残留成纤维细胞所含的HCMV 通过定量反转录PCR( RT - PCR) 扩增脯-4-羟化酶exon12a mRNA 来校正。结果显示, 使用去白细胞滤器后, 每单位( 200 ml) 全血血液中, 白细胞的去除率为99. 98% 。全血中平均HCMV 病毒载量从3 742 拷贝/ μl 降至22. 57 拷贝/ μl, 降低2. 50 log10 , 说明去白细胞滤器虽然可明显减少全血中HCMV 病毒的载量, 但并不能完全去除病毒。     

氧化苦参碱对K562肿瘤细胞增殖的影响

目的:研究氧化苦参碱(OM)对人白血痛细胞系K562生长增殖的影响.方法:运用MTT比色法、活细胞计数法、集落形成法以及透射电镜观察检测OM对人白血病细胞系K562增殖抑制作用.结果:MTT实验、生长曲线及集落形成实验显示OM能明显抑制K562细胞的增殖.随着OM浓度的增加,K562细胞存活细胞显著降低,呈现明显的刺量依赖性,经相关分析,细胞抑制率与OM浓度呈正相关(r=0.9010),其半数抑制浓度(IC50)为0.33 mg/ml.透射电镜下显示在低浓度即有明显的诱导细胞凋亡的作用,出现核固缩、核碎裂、凋亡小体等典型的凋亡形态.结论:OM具有抑制K562白血病细胞增殖诱导肿瘤细胞凋亡的作用.