miR-769-3p 对甲型 H1N1流感病毒核蛋白表达的调控

目的:预测靶向甲型流感病毒核蛋白(NP)基的微小 RNA(miRNA),并检测其对 NP 表达的影响.方法:从miRBase 数据库中获取人成熟 miRNA 序列,利用 miRanda 软件预测潜在靶向流感病毒 A/FM/1/47(H1N1) NP 基的人 miRNA;通过双萤光素酶报告基系统及 Western 印迹验证所预测的 miRNA 对 NP 表达的影响.结果:用 miRanda软件在流感病毒 A/FM/1/47(H1N1) NP 基上预测得到分值及最小结合自由能均较好的 miR-769-3p;双萤光素酶报告基结果显示 miR-769-3p 能显著降低报告基载体萤光素酶的表达;Western 印迹结果显示 miR-769-3p 能明显抑制 NP 的表达,但突变 NP 基上的 miR-769-3p 结合位点后,miR-769-3p 不能抑制 NP 的表达.结论:miR-769-3p 可靶向流感病毒 A/FM/1/47(H1N1) NP 基并抑制 NP 的表达,为抗甲型流感病毒的 miRNA 药物研发提供了据和潜在药物靶标.     

S100A4原核表达载体的构建以及蛋白的表达和纯化

S100A4是S100蛋白家族的成员,在细胞的增殖、分化、损伤修复以及肿瘤细胞转移等方面发挥重要的调控作用.本研究将S100A4全长基因构建到pET28a原核表达载体上,利用大肠杆菌表达系统表达和纯化出高纯度的重组人S100A4.通过试验证明,重组人S100A4蛋白在体外可以有效地增强黑色素瘤细胞A375-S2的增殖.重组人S100A4原核表达与纯化方法的建立将促进其结构和生物学功能研究,并且对于S100蛋白家族其它蛋白的表达与纯化具有重要的参考意义.     

红光熊蜂卵黄原蛋白基因的cDNA全长序列克隆和表达分析

红光熊蜂Bombus ignitus Smith是许多经济作物和野生植物的重要授粉昆虫之一。卵黄原蛋白基因(vitellogenin,Vg)在昆虫的生殖调控中和行为方面起到重要的作用,本试验对Vg基因全长cDNA的克隆和测序及在蜂王、工蜂和雄性蜂三型蜂中的表达分析得出:Vg基因的全长cDNA为5 481 bp,GenBank中的登录号为FJ913883,有一个完整的开放阅读框(ORF),编码1 772个氨基酸,N-末端的前16个氨基酸为一个信号肽。接近C-末端区域存在保守的GL/ICG基元,其后含有9个半胱氨酸,而且DGXR位于GL/ICG基元上游18个氨基酸残基处。其氨基酸序列与韩国的熊蜂B.ignitus和B.hypocrita相似性高达95%,与西方蜜蜂Apis mellifera的相似性达到51%。Vg的mRNA首先在蜂王蛹期的白眼蛹(Pw)时期出现,其表达量在蜂王整个蛹期发育过程中呈上升趋势,且在黑眼蛹(Pbd)时期达到最高,在成年蜂的脂肪体中的表达量仍在升高,甚至更高。Vg也在工蜂蛹期的白眼蛹(Pw)时期被检测到,然后在整个蛹期发育过程中呈现上升趋势,在刚羽化出房时达到高峰,Vg的mRNA水平随着成年蜂日龄的增加而增加,到15日龄时达到最高,然后呈现下降趋势。对于雄性蜂,Vg的mRNA虽然卵黄原蛋白基因的mRNA水平几乎在整个蛹期发育阶段都表达,但是表达水平非常低,只有在刚羽化出房时期表达水平较高。     

甘肃麦积山风景区熊蜂物种多样性调查

熊蜂属昆虫是山区植物的重要传粉者,在保持生物多样性和维护生态系统平衡方面发挥着十分重要的作用。麦积山风景区位于甘肃东南部的西秦岭山区,地处北部温带黄土高原向南部亚热带河谷山地和西部高寒青藏高原过渡的地带,地理位置独特,生物资源丰富。为了探明该景区内熊蜂物种多样性现状,笔者于2007—2010连续4年对该景区的熊蜂属昆虫进行了系统调查。结果表明:2007—2010年,在该景区12个采样点共采集熊蜂属昆虫1 765头,隶属于7亚属20种;在12个采样点中,石门和净土寺的熊蜂种类丰富度最高;在20种熊蜂中,火红熊蜂Bombus pyrosoma、红光熊蜂B.ignitus、疏熊蜂B.remotus、小峰熊蜂B.hypocrita s.l.和重黄熊蜂B.picipes的物种多度较高,依次位居前5位;麦积山风景区熊蜂区系成分以东洋区+古北区共有种为主(55%),其中东洋区成分高于古北区成分;麦积山熊蜂区系成分和南部亚热带湿润气候区的白水江自然保护区以及西部高寒气候区的扎尕那自然保护区比较接近,和北部温带半干旱气候区的崆峒山风景区相距较远。     

笃斯越桔化学成分及其功能活性的研究进展

笃斯越桔(Vaccinium uliginosum)因其富含花青苷等生物活性成分而备受关注。迄今为止, 从笃斯越桔果实中已鉴定出24种花青苷、32种黄酮醇苷、24种有机酸、28种芳香成分及少量儿茶素及其低聚物。同时, 笃斯越桔还含有丰富的矿质元素、维生素及氨基酸等营养成分。大量的体内和体外实验结果表明, 笃斯越桔具有强大的抗氧化、抗衰老、抗炎、抗癌、提高记忆力、预防心血管疾病和保护视力等生理活性。该文介绍了笃斯越桔资源分布情况及其遗传背景, 综述了其果实和叶片中的主要功能成分、营养物质及芳香成分的组成和含量, 讨论了遗传和环境等因素对其活性物质合成和积累的影响, 以期为其新品种选育及综合利用提供参考依据。     

Cloning,Characterization and Primary Function Study of a Novel Gene,Cymg1,Related to Family 2 Cystatins

Cystatins are cysteine proteinase inhibitors,We found two expression sequence tags (ESTs),CA463109 and AV042522,from a mouse testis library using Digital differential display (DDD).By electricalhybridization,a novel gene,Cymgl(GenBank accession No.AY600990),which has a full length of 0.78 kb,and contains four exons and three introns,was cloned from a mouse testis eDNA library.The gene is locatedin the 2G3 area of chromosome 2.The full eDNA encompasses the entire open reading frame,encoding 141amino acid residues.The protein has a cysteine protease inhibitor domain that is related to the family 2cystatins but lacks critical consensus sites important for cysteine protease inhibition.These characteristicsare seen in the CRES subfamily,which are related to the family 2 cystatins and are expressed specifically inthe male reproductive tract.CYMG1 has a 44%(48/108)identity with mouse CRES and 30%(42/140)identity with mouse cystatin C.Northern blot analysis showed that the Cymgl is specifically expressed inadult mouse testes.Cell location studies showed that the GFP-tagged CYMG 1 protein was localized in thecytoplasm of HeLa cells,lmmunohistochemistry revealed that the CYMG1 protein was expressed in mousetestes spermatogonium,spermatocytes,round spermatids,elongating spermatids and spermatozoa.RT-PCRresults also showed that Cymgl was expressed in mouse testes and spermatogonium.The Cymgl expressionlevel varied in different developmental stages:it was low 1 week postpartum,steadily increased 2 to 5 weekspostpartum,and was highest 7 weeks postpartum.The expression level at 5 weeks postpartum was main-tained during 13 to 57 weeks postpartum.The Cymgl expression level in the testes over different develop-mental stages correlates with the mouse spermatogenesis and sexual maturation process.All these indicatethat Cymgl might play an important role in mouse spermatogenesis and sexual maturation. Cystatins are cysteine proteinase inhibitors,We found two expression sequence tags(ESTs),CA463109 and AV042522,from a mouse testis library using Digital differential display (DDD).By electricalhybridization,a novel gene,Cymgl(GenBank accession No.AY600990),which has a full length of 0.78 kb,and contains four exons and three introns,was cloned from a mouse testis eDNA library.The gene is locatedin the 2G3 area of chromosome 2.The full eDNA encompasses the entire open reading frame,encoding 141amino acid residues.The protein has a cysteine protease inhibitor domain that is related to the family 2cystatins but lacks critical consensus sites important for cysteine protease inhibition.These characteristicsare seen in the CRES subfamily,which are related to the family 2 cystatins and are expressed specifically inthe male reproductive tract.CYMG1 has a 44%(48/108)identity with mouse CRES and 30%(42/140)identity with mouse cystatin C.Northern blot analysis showed that the Cymgl is specifically expressed inadult mouse testes.Cell location studies showed that the GFP-tagged CYMG 1 protein was localized in thecytoplasm of HeLa cells,lmmunohistochemistry revealed that the CYMG1 protein was expressed in mousetestes spermatogonium,spermatocytes,round spermatids,elongating spermatids and spermatozoa.RT-PCRresults also showed that Cymgl was expressed in mouse testes and spermatogonium.The Cymgl expressionlevel varied in different developmental stages:it was low 1 week postpartum,steadily increased 2 to 5 weekspostpartum,and was highest 7 weeks postpartum.The expression level at 5 weeks postpartum was main-tained during 13 to 57 weeks postpartum.The Cymgl expression level in the testes over different develop-mental stages correlates with the mouse spermatogenesis and sexual maturation process.All these indicatethat Cymgl might play an important role in mouse spermatogenesis and sexual maturation.     

山西省熊蜂属区系调查(膜翅目,蜜蜂科)

作者在2005～2007年对山西省熊蜂属的区系进行了调查.根据所采集的标本和中国科学院动物研究所馆藏标本,经鉴定该地区有熊蜂属10亚属28种,其中山西省新纪录种7种.另外,作者对该地区的熊蜂区系成分进行了初步的分析.     

河北地区熊蜂物种多样性与蜂群繁育特性

基于2005-2009年从河北地区（包括河北省、北京市和天津市）所采集的1893号熊蜂标本资料,结合中国科学院动物研究所馆藏记录,分析了河北地区熊蜂物种多样性和蜂群繁育特性. 结果表明：河北地区共有熊蜂属昆虫8亚属32种,其中河北省32种,北京市18种,天津市5种;西部太行山区、北部燕山山区和坝上高原地区熊蜂种类丰富度和多度较高;河北地区熊蜂的访花植物涉及到21科80种,其中,菊科、豆科和唇形科植物是大多数熊蜂种类访问的主要对象;小峰熊蜂、红光熊蜂、密林熊蜂、火红熊蜂和重黄熊蜂5种熊蜂群势强大,平均单群蜂的工蜂数量在110只以上,雄蜂数量在160只以上,子代蜂王数在30只以上;这5种熊蜂的繁育成群率均在50%以上,易于人工驯养,具有重要的传粉利用价值.     

Fusion Protein of Interleukin 4 and Diphtherial Toxin with High Cytotoxicity to Cancer Cells

One means to improve the selectivity of cancer therapyis by directing foreign protein with activity againsttherapeutic targets that displays different expression levelon malignant cells from normal cells. There have hadsubstantial efforts to rationally de…     

血管内皮细胞生长因子及其受体在IgA肾病患者肾组织中表达的研究

目的　检测 2 4例不同病理改变的IgA肾病患者肾组织中血管内皮细胞生长因子 (Vascularendothelialgrowthfactor,VEGF)及其受体VEGF R2 (flk 1)和Ⅳ型胶原的表达 ,并将其表达水平与临床病理改变进行相关分析。方法　应用免疫组织化学SP法。结果　VEGF主要表达于肾小球脏层上皮细胞 ,flk 1主要表达于肾小球和间质血管内皮细胞。肾小球内VEGF和flk 1表达呈正相关 (P <0 0 5 ) ,两者在中度病变组肾小球内表达最多 ,重度病变组表达最少 (P <0 0 5 )。随病理改变程度加重 ,患者 2 4h尿蛋白定量、血压、血肌酐水平均明显增高 (P <0 0 5 )。肾小球内VEGF表达在轻、中度病变组与尿蛋白水平呈正相关 (P <0 0 5 ) ;在重度病变组则无相关性 ,与患者血压、血肌酐水平也无明显相关性 (P >0 0 5 )。肾小球内Ⅳ型胶原随病变加重表达增多 ,在重度病变组表达最多 (P <0 0 1) ;在轻、中度病变组肾小球内VEGF表达与Ⅳ型胶原的表达呈正相关 (P <0 0 5 )。各组肾小球内VEGF表达与肾小球微血管密度呈正相关 (P <0 0 1) ,重度病变组肾小球微血管密度减少 (P <0 0 5 )。结论　这些结果说明VEGF在IgA肾病的病理进展过程中发挥重要作用 ,可能涉及内皮细胞通透性增强、细胞外基质合成增多、血管生成障碍等多种机制