Cortical gene expression pattern in rat focal cerebral ischemia

AIM: To investigate the genes differential expression in cortex during rat focal cerebral ischemia.METHODS: cDNA microarray chips containing numerous cDNAs were used to investigate the gene expression pattern between samples of focal cerebral ischemia and sham-control operation rats. RESULTS: Two hundred and eleven genes differentially expressed were screened out, among these genes, up-and down-regulated genes were 199 and 12, respectively. CONCLUSIONS: The analysis of gene expression pattern of focal cerebral ischemia based on cDNA microarray can realize high-throughput screening of the genes associated with the focal cerebral ischemia. The differential expression of genes may be related to the pathogenesis of focal cerebral ischemic diseases.     

Inhibition of K562 cell growth by antisense drug targeting with VEGF mRNA in vitro

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs.     

Comparative expression profile of microRNAs and piRNAs in three ruminant species testes using next‐generation sequencing

microRNA (miRNA) and piwi‐interacting RNA (piRNA) are two classes small non‐coding regulatory RNAs that play crucial roles in multiple biological processes such as spermatogenesis. However, there are no published studies on conjoint analysis of miRNA and piRNA profiles among cattle, yak and their interspecies (the dzo) using sequencing technology. Next‐generation sequencing technology was used to profile miRNAs and piRNAs among those three ruminants to elucidate their functions. A total of 119, 14 and six differentially expressed miRNAs were obtained in cattle vs. dzo, cattle vs. yak and yak vs. dzo comparison groups, while there were 873, 1,065 and 1,158 differentially expressed piRNAs in those three comparison groups. The expression of three miRNAs was validated in the three ruminants, and the results suggested that the miRNA expression profiles data could represent actual miRNA expression levels. Moreover, the putative targets of differentially expressed miRNAs were predicted by their own genome, it is worth to note that both the cattle and yak genome were used for dzo, then the targets were subjected to GO enrichment and KEGG pathway analysis, revealing the likely roles for these differentially expressed miRNAs in spermatogenesis. In conclusion, this study provided a useful resource for further elucidation of the miRNAs and piRNAs regulatory roles in spermatogenesis. It may also facilitate the development of therapeutic strategies for dzo reproduction research.     

低温胁迫对细茎柱花草与TPRC2001-1柱花草抗寒生理指标的影响

为了探究柱花草的抗寒性,本研究以细茎柱花草(Stylosanthes guianensis var.intermedia)和TPRC2001-1柱花草(S.guianensis‘TPRC2001-1’)营养期叶片为材料,分别进行时长为0(对照)、24h和48h的低温处理,然后对其相对电导率、丙二醛、脯氨酸、可溶性糖以及可溶性蛋白的含量等指标进行测定与分析。结果表明,两种柱花草叶片的抗寒性生理指标随着低温处理时间的增加逐步上升。低温处理后,细茎柱花草的相对电导率和丙二醛含量显著低于TPRC2001-1柱花草(P0.05),而脯氨酸、可溶性糖和可溶性蛋白的含量显著高于TPRC2001-1柱花草(P0.05)。综合各项生理指标可得,两种柱花草的抗寒性强弱为细茎柱花草TPRC2001-1柱花草。     

假臭草幼苗生长对水氮耦合效应的响应

为揭示入侵植物假臭草(Praxelis clematidea)幼苗生长对不同水、氮养分耦合模式的响应机制,探讨其与入侵性的关系,以假臭草为供试材料,设计3个供水量水平、4个供氮量水平共12个处理,比较假臭草的形态、生物量分配和叶绿素含量等生长指标对不同水氮耦合效应的可塑性反应。结果显示,假臭草幼苗生长受水氮耦合影响显著(P0.05),在适宜水分和养分条件,如中水中氮处理(沙土饱和含水量60%~70%,供水量60 mL·kg~(-1),供氮量0.30g·kg~(-1))处理下,假臭草幼苗株高、茎粗、叶面积、总根长度和根表面积等根、茎、叶形态指标表现较好,生物量积累较多,植株生长较好。低水处理(沙土饱和含水量30%~40%,供水量30 mL·kg~(-1))和高水处理(沙土饱和含水量90%~100%,供水量90mL·kg~(-1))的各个氮含量处理的假臭草植株的各项根、茎、叶形态及生物量指标均差于中水处理。假臭草叶绿素对氮素较为敏感,随着氮素增加其积累量也相应增多。研究结果表明,假臭草入侵植物喜好一定的水、氮环境,过多或不足的水分或养分则不利于假臭草的生长发育,但仍有一定的分布,这与其入侵性密切相关。     

羊种布鲁氏菌疫苗株PCR-RFLP鉴定方法的建立

为建立一种羊种布鲁氏菌Rev.1疫苗株PCR-RFLP鉴定方法,利用羊种布鲁氏菌Rev.1具有链霉素抗性的特性,选取与链霉素抗性相关编码核糖体蛋白S12的rps L基因为模板设计引物,经PCR扩增后,将产物进行Nci I酶切,发现羊种布鲁氏菌疫苗株Rev.1出现约500 bp条带,而不具有链霉素抗性的羊种布鲁氏菌株16M则出现384 bp条带。结果表明,PCR-RFLP方法能够用于羊种布鲁氏菌Rev.1疫苗株的鉴定,这为今后区分羊种布鲁氏菌疫苗株Rev.1免疫与野株感染提供了基础。     

杏鲍菇多孢分离菌株ISSR、RAPD分子标记及体细胞不亲和性分析

通过杏鲍菇多孢分离菌株的ISSR、RAPD分子标记和体细胞不亲和性分析,体细胞不亲和性分析得出子代杏1、杏3、杏4与出发菌株湘杏98有拮抗线,杏1、杏2、杏3、杏4四株子代均有明显的拮抗线。16条ISSR引物和19条RAPD引物分别扩增出99、154个条带,多态性条带分别占48.48%、61.00%;相似系数变异范围分别为0.63~0.86、0.57~0.83。湘杏98、杏2、杏3在2种标记的聚类结果上存在差异。体细胞不亲和性分析从表型上(拮抗线有无)反应了菌株的遗传变异。RAPD标记遗传相似系数小于ISSR标记,表明ISSR标记更适合亲缘关系较近的种群间遗传多样性分析。     

苦瓜转录因子McEIL2的克隆及其表达分析

本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。     

鱼肠道弧菌免疫检测方法的研究

采用灭活的鱼肠道弧菌作为抗原,制备兔抗血清,建立了鱼肠道弧菌的间接荧光抗体检测技术(indirect im-munofluorescence assay test,IFAT)、Western-blotting免疫印迹检测技术。IFAT结果显示阳性信号覆盖整个菌体,明亮,清晰,荧光信号呈长弧形且两端钝圆,与鱼肠道弧菌形状一致。Western-blotting结果显示共10条蛋白带发生免疫反应,其中6条蛋白带结果较清晰,显示出较强的特异性免疫反应,阴性对照组无条带。IFAT和Western-blotting结果说明所建立检测方法能够准确、快速的检测出鱼肠道弧菌。     

从文化视角看中国古典园林

文章从文化的角度,对中国古典园林所蕴含的哲学思想及审美情趣进行了阐述,探讨了中国传统文化对园林艺术的影响并分析中国传统园林中意境形成的原因及表现方法,可以获得对其更深的认识,以期给现代设计师以启发和思考,从传统文化中汲取精华走一条中国特色的设计之路.