Characterization and the broad cross‐species applicability of 20 anonymous nuclear loci isolated from the Taiwan Hwamei (Garrulax taewanus)

We isolated 20 anonymous nuclear loci (8556 bp in total) from the Taiwan Hwamei (Garrulax taewanus), an endemic songbird of Taiwan. A panel of nine to 15 individuals with unknown relationship was used to characterize polymorphism of these loci. We identified 46 single nucleotide polymorphic sites (SNPs) in 15 polymorphic loci. Frequency of SNPs was one per every 186 bp in average. Nucleotide diversity, θ, ranged from 0.00054 to 0.00371 per locus. We also tested cross‐species applicability of these loci on 17 species from eight different passerine families. All 20 loci could be successfully amplified (ranged from one to 16 species, mean = 7.9 species).     

地塞米松当归多糖前体药在大鼠胃肠道内容物中的释药特性

目的:探讨以当归多糖为载体的地塞米松前体药在大鼠胃肠道内容物中的释药特性,考察其结肠靶向性。方法:将前体药与大鼠胃肠道不同部位内容物稀释液于37℃共同孵育,不同时间点取样,高效液相色谱法检测释放的地塞米松(dexamethasone, DEX)及地塞米松琥珀酸单酯(dexamethasone hemisuccinate,DSH)。结果:前体药在大鼠胃肠道各部位内容物稀释孵育液中均有DEX及DSH释放;DEX在小肠近端的释放量最大,在结肠和盲肠中的释放量次之,在胃和小肠远端的释放量较小;DSH在结肠和盲肠中的释放量大,在胃、小肠近端和小肠远端释放量小;在160min孵育时间时,DEX和DSH在结肠和盲肠中的释放总量是在胃、小肠近端和小肠远端释放总量的1．95倍。结论:以当归多糖为载体的地塞米松前体药具有一定的结肠靶向释药特性,有可能用作局部治疗结肠炎的结肠靶向药物。     

CDC6靶向RNA干扰质粒载体的构建及生物活性鉴定

目的:构建小鼠CDC6基因的RNAi真核表达载体PGCsilencer TM u6/Neo/GFP/RNAi,观察其转染小鼠肝细胞前后CDC6的表达变化。方法:根据GenBank中CDC6的序列,设计特异性siRNA序列,将模板序列克隆至PGCsilencer U6/Neo/GFP质粒中,通过测序鉴定后,用脂质体将重组子转染至正常小鼠肝细胞中,用RT-PCR检测CDC6的mRNA的表达及用Western blot方法检测CDC6蛋白水平的表达,并比较转染前后其表达水平的变化。结果:经测序,模板序列与设计序列完全正确,经过RT-PCR及Western blot方法检测,转染干扰质粒后,小鼠肝细胞中CDC6表达在mRNA及蛋白水平都有明显的下降。结论:成功构建了CDC6基因的RNAi真核表达载体并转染至小鼠肝细胞中,为下一步探讨CDC6在肝再生的作用奠定了基础。     

细胞间隙连接通讯与肿瘤

由连接蛋白构成的细胞间隙连接通讯(GJIC)广泛存在于脊椎动物中,可以直接介导细胞间小分子物质的传递,而不需要通过细胞间质。GJIC与肿瘤密切相关,多数肿瘤GJIC能力降低或丧失,连接蛋白不表达或胞内定位,而恢复GJIC可以抑制肿瘤, GJIC已成为肿瘤预防与治疗研究的潜在靶点之一。     

黄鳝生殖干细胞在性逆转生殖发育过程中的变化

应用组织学方法及免疫组织化学技术显示,黄鳝性逆转生殖发育过程中,生殖干细胞(GSCs)定位分布于生殖褶中,黄鳝雌性发育阶段的GSCs分散或成团存在,间性及雄性发育阶段GSCs均区分为A、B两种不同类型,雌性发育阶段GSCs与A、B两类GSCs在超微结构上存在差异。结果表明,生殖褶中GSCs是黄鳝分化生殖腺中唯一具有有丝分裂能力的生殖细胞群,雌性发育阶段GSCs表现出卵原干细胞特征,间性及雄性发育阶段GSCs为精原干细胞。CD49整合素是黄鳝雌性发育阶段GSCs和A类GSCs的表征分子。     

Correlation between haplotype of apolipoprotein B gene and natural longevity persons in Uygur Nationality

This paper investigated the correlation between polymorphisms and haplotypes in the apolipoprotein B (apoB) gene (SP-I/D, XbaI-RFLP, VNTR) and natural longevity persons among the Uygur people in Xin-jiang. For this purpose, 191 healthy Uygur individuals aged above 90 from Hetian area of Xinjiang were recruited, and another 53 persons aged 65—70 from the same nationality, the same region and with the same gender ratio, served as the control group. Genotyping was performed by PCR-SSP, PCR-RFLP and PCR-sequencing methods. Logistic regression analyses revealed that the frequencies of X X genotype, M and L alleles and the genetypes composed of M and L were significantly higher in the longevity group than in the control group. In haplotype analyses, we found that, in the long-lived people, the frequency of haplotypes composed of the X and M alleles was significantly higher whereas the frequency of haplotypes composed of the X- and S alleles was significantly lower (both P<0.05) I than those of their controls. These results indicated that the S allele, SS genotype and X -S, D-S, D-X -S haplotypes were the possible adverse factors, whereas the M, L alleles, X X , MM, ML, LL genotypes and I-X -M, X -M haplotypes were the possibe protective factors for the naturally long-lived Uygur people in China.     

Structure comparisons of Aedes albopictus densovirus with other parvoviruses

Parvoviridae is a family of the smallest viruses known with a wide variety of hosts. The capsid structure of the Aedes albopictus C6/36 cell densovirus (C6/36 DNV) at 1.2-nm resolution was obtained by elec-tron cryomicroscopy (cryoEM) and three-dimensional (3D) image reconstruction. Structure compari-sons between the C6/36 DNV and other parvoviruses reveal that the degree of structural similarity be-tween C6/36 DNV and the human parvovirus B19 is higher than that between C6/36 DNV and other in-sect parvoviruses. The amino acid sequence comparisons of structural and non-structural proteins also reveal higher levels of similarity between C6/36 DNV and parvovirus B19 than those between C6/36 DNV and other parvoviruses. These findings indicate that C6/36 DNV is closely related to the human virus B19, and the former might evolve from the human species other than from other insect viruses.     

慢病毒介导的RNA干涉对乳腺癌SKBR3细胞HER2受体的下调及生长抑制

大约30％的乳腺癌中有表皮生长因子受体家族蛋白HER2的过表达,此类癌症的预后差,恶性程度高。RNA干涉（RNAi）是最近发展起来能特异性抑制哺乳动物细胞中基因表达的新技术。本文在以往获得的能够产生良好基因沉默效应的小干涉RNA（siRNA）的基础上,构建了U6和H1双启动子siRNA表达载体,并转染HER2高表达乳腺癌SKBR3细胞定量测定了其HER2下调效应。随后,siRNA表达盒经LR重组反应被克隆入慢病毒载体中,在成功包装成病毒后,感染SKBR3并经荧光定量PCR、蛋白印迹杂交和流式细胞仪一系列实验证明慢病毒介导的RNAi确实能有效地下调肿瘤抗原HER2的表达。细胞长期增殖实验表明经慢病毒处理后细胞生长得到抑制。我们的研究为进一步阐明HER2与癌症恶化的关系以及发展新的基因治疗药物提供了工具和可能。     

毛果杨nsLTP基因家族全基因组水平鉴定及其表达特性分析

植物非特异性脂质转移蛋白（non-specific lipid transfer proteins,nsLTP）是一类多基因家族编码碱性蛋白,负责脂肪酸体外结和与膜之间的磷脂转移,在植物生长发育和逆境胁迫响应中扮演着重要角色。目前为止,尚无模式植物毛果杨（Populus trichocarpa）nsLTP家族的研究报导。本研究从全基因组水平对PtrnsLTP家族成员的基因数量、亲缘关系、基因结构、编码蛋白保守基序等特性进行了分析,结果表明：PtrnsLTP家族共由39个基因组成,进化成5个亚家族,其中A亚族含有6个基因、B亚族含有2个、C亚族含有13个、D亚族含有3个、E亚族含有15个。PtrnsLTP家族包含7对旁系同源基因,其中1对大于1,6对Ka/Ks均远小于1,且这6对基因均处于同一个大的进化分支上,进化压力的不同导致基因间的功能出现了分化,编码蛋白均含有Motif 1和 Motif 2保守基序。利用qRT-PCR技术并结合杨树转录组数据对PtrnsLTP的组织表达与盐胁迫响应特性研究发现：各家族成员在毛果杨根、茎和叶中均有表达且经qRT-PCR技术验证后与网站预测结果基本吻合,有11、15和13个成员分别在根、茎和叶中有较高的表达,表明该基因家族参与了杨树不同组织的生长发育;NaCl胁迫下,该家族39个基因中分别有26个成员在根部、14个成员在叶部表达量随着胁迫时间的增加而升高,而32个基因在茎部表现为先升高后降低的趋势。本研究结果对于PtrnsLTP家族基因生物学功能的鉴定与盐胁迫响应基因资源的工作有着积极的推动作用。     

革兰氏阴性菌TonB依赖性受体的功能与结构研究进展*

为维持生长所需,革兰氏阴性菌需要从外界摄取多种营养物质。分子量小于600 Da的分子可以通过自由扩散的方式通过革兰氏阴性菌的外膜,而大分子物质则需要特殊的转运系统才能将其转运至革兰氏阴性菌的胞内。革兰氏阴性菌对大分子营养物质的识别和转运主要由TonB依赖性受体负责完成。所有革兰氏阴性菌中均有TonB依赖性受体的存在,然而不同种类的革兰氏阴性菌拥有TonB依赖性受体的数量不同且功能各异。最近研究表明,TonB依赖性受体不仅参与了铁、血红素、锰、锌、镍、维生素、碳水化合物等多种营养物质的摄取,而且参与了蛋白酶的分泌。为对TonB依赖性受体提供更为深入和系统的理解,详细介绍了目前已知的TonB依赖性受体的功能及结构,以期为更进一步探知TonB依赖性受体未知功能提供可参考依据。