分离MEF和ES细胞的差速贴壁法研究(简报)

胚胎干细胞的分化控制是胚胎干细胞研究的一个重要方面。由于常规的拟胚体诱导途径是在形成拟胚体后才开始进行诱导分化,受多种胚层的共同作用使得我们无法简便探索诱导分化的机制。而且用这种方法进行的诱导分化试验的结果检测比     

水蒸汽蒸馏巴柑檬叶和果皮精油化学成分的研究

巴柑檬是我们第一次从国外引种成功的一种名贵香料植物。用色谱-质谱-计算机联用技术、毛细管气相色谱保留指数法和标准品叠加法分析了水蒸汽蒸馏巴柑檬叶和果皮精油的化学成分。从叶和果皮精油分离出来的220个和200个成分中,分别鉴定出48个和57个成分。鉴定组分的含量分别占叶和果皮精油的99.80％和98.54％。叶精油与果皮精油在化学成分方面的主要区别是叶油中萜烃化合物含量较低,而萜醇类化合物含量较高。     

云南呈贡梁王山现代花粉雨的研究

本文通过对云南呈贡梁王山5块表土分析,初步研究了主要植物花粉的百分含量与其植物覆盖率之间的数量关系,并用校正系数R值表示。按照R值的大小,分为两组:R>1属于超代表性,包括有松、桤木、马桑、蒿和部分蕨类植物;R<1属于低代表性,包括有油杉、栲和石栎、滇青冈、栎、铁仔。在分析松粉分布特征基础上,认为昆明地区西风急流对松粉的传播是主要因素。     

波形蛋白介导猪繁殖与呼吸综合征病毒感染Marc-145细胞作用的初步研究

为了研究波形蛋白在介导PRRSV感染过程中的作用,根据GenBank中已发表的波形蛋白序列,设计特异性引物,利用RT-PCR方法从Marc-145细胞中扩增目的基因,克隆入pET-28a,转化表达菌BL21(DE3)中进行诱导表达,用SDS-PAGE和Western blot鉴定表达产物,将表达产物纯化后免疫BALB/c小鼠制备血清,用病毒阻断实验验证PRRSV与波形蛋白及其抗体的关系。用ELISA方法确定了重组波形蛋白与PRRSV结构蛋白N蛋白和囊膜蛋白GP5蛋白之间的关系。结果表明,成功的从Marc-145细胞中扩增出全长的波形蛋白基因,将其克隆入pET-28a,诱导后得到高效表达,表达蛋白纯化后免疫小鼠抗体效价达到105,病毒阻断实验表明波形蛋白能部分阻断PRRSV感染,其抗血清能完全阻断PRRSV感染。ELISA结果表明波形蛋白能与N蛋白结合而不与GP5蛋白结合。此结果为PRRSV感染的细胞的受体机制增添了新的内容,为PRRSV感染过程中受体间的相互作用关系研究奠定基础。     

Twist及Akt2在甲状腺乳头状癌中的表达及意义

目的探讨Twist、Akt2在甲状腺乳头状癌中的表达和相互关系。方法应用免疫组化方法,对60例甲状腺乳头状癌、10例结节性甲状腺肿进行Twist、Akt2表达的研究。结果甲状腺乳头状癌Twist及Akt2阳性表达率分别为81.67%(49/60)及60.00%(36/60),结节性甲状腺肿Twist及Akt2阳性表达率分别为0.00%(0/10)及10.00%(1/10),前者与后者相比差异有统计学意义(P<0.05);Twist及Akt2在甲状腺乳头状癌中的表达水平与病人的年龄及原发肿瘤分期无关,但与有无淋巴结转移相关(P<0.05),且Akt2的表达在T3,4期肿瘤及T1,2期肿瘤相比也具有显著性(P<0.05);Twist及Akt2的表达呈正相关(r=0.492,P=0.000)。结论在大多数甲状腺乳头状癌的上皮细胞中都存在Twist及Akt2的过表达,其淋巴结转移的发生与Twist及Akt2过表达或功能的不正常可能有密切的关系;Twist与Akt2的表达呈正相关。     

重组腺相关病毒载体相关性杂质

可以稳定表达治疗基因而无明显不良反应的重组腺相关病毒(rAAV)载体被认为是最有发展前景的基因治疗载体。但如何建立可以有效去除rAAV载体内具有潜在致病危害的杂质、产品质量符合临床使用要求的纯化工艺是研究人员面临的巨大挑战。其中针对载体相关性杂质的纯化工艺尤为关键,因为该类杂质的性质与真正的rAAV载体极其相似,一旦存在便难以去除,且会引起严重不良反应。以下总结了该类杂质形成的过程及有别于rAAV载体的特点,并对可以防止其生成或将其与rAAV载体有效分离的技术手段进行了评价。     

越冬和复苏时期太湖水体蓝藻群落结构的时空变化

为研究太湖蓝藻在越冬与复苏时期群落结构的时空变化规律,于2008年11月,2009年2月,2009年4月,在太湖富营养化较严重的湖区选取8个采样点 (梅梁湾、竺山湾、贡湖湾、大浦、西太湖、南太湖、湖心和湖湾交汇处),分3层采水样,过滤并提取样品DNA经PCR扩增蓝藻16S rDNA序列,采用T-RFLP(末端标记的限制性酶切片段长度多样性)技术分析蓝藻群落结构和多样性变化。共得到87个不同的T-RFs(末端限制性酶切片段),表明太湖蓝藻具有丰富的基因多样性。T-RF相对丰度和聚类分析结果表明,太湖蓝藻群落结构在垂直空间上相似性较高,相似度 > 50%;在水平空间,与Microcystis spp.对应的信号峰在8个采样点均为最强峰(相对丰度为17.7% 47.5%)。竺山湾蓝藻多样性最低,西太湖最高,但其余采样点间蓝藻群落和Shannon多样性指数没有显著差异(P>0.05)。Microcystis相对丰度与Shannon多样性指数呈显著负相关(皮尔逊相关系数为-0.958)。在时间尺度上,相似性分析(Analysis of similarity,ANOSIM)结果显示太湖蓝藻群落结构存在极显著差异( P<0.01) 。春季复苏时蓝藻多样性最高,秋季衰亡时最低。聚类分析表明样品聚成两大特征类群,秋季衰亡时样品独自聚为1支,而春季复苏期和冬季越冬期样品彼此混杂。     

Design,Synthesis, Biological Activity Evaluation and Action Mechanism of Myricetin Derivatives Containing Thiazolebisamide

The plant diseases caused by a variety of pathogens such as viruses, bacteria and fungi pose a great threat to global food production and food safety. Therefore, the search for green, efficient and pollution-free pesticides has become an important task. In this article, 23 myricetin derivatives containing thiazolebisamides active groups have been designed and synthesized. Their activities were evaluated by performing in vitro antibacterial and in vivo antiviral assays, microscale thermophoresis (MST) and molecular docking assays. The results of in vivo antiviral assays showed that compounds A4 and A23 exhibited good antiviral activity with EC₅₀ values of 79.0 and 54.1 μg/mL for therapeutic activity and 103.3 and 91.2 μg/mL for protective activity, respectively. The dissociation constants (Kd) values of compounds A4 and A23 against TMV-CP were 0.021 and 0.018 μM, respectively, determined by microscale thermophoresis (MST), which were much smaller than those of the commercial drug ningnanmycin (NNM), which were 2.84 μM. The interaction of compounds A4 , A23 with TMV-CP was further verified at the molecular level. In addition, in vitro antifungal assays of this series of compounds showed that they exhibited some inhibitory activity against a variety of fungi, especially against the phytophthora capsici. Among them, A13 and A20 showed similar inhibitory activity to the control drug azoxystrobin at 100 μg/mL against the phytophthora capsici.     

Isolation,characterization and cytotoxic activity of benzophenone glucopyranosides from Mahkota Dewa (Phaleria macrocarpa (Scheff.) Boerl)

Two benzophenone glucopyranosides have been isolated from the nut shell part of Mahkota Dewa. The structures were identified as 2,4′,6-trihydroxy-4-methoxy-benzophenone-2-O-β-d-glucoside (Mahkoside A) and 2,4′,6-trihydroxy-4-methoxy-6″-acetyl-benzophenone-2-O-β-d-glucoside (Mahkoside B). Mahkoside B was recognized as a novel compound. Furthermore, a series of benzophenone glucopyranoside derivatives (compounds 3–18) were synthesized and their bioactivities were characterized. Our results demonstrated that compound 18 has significant cytotoxicity against two esophageal cancer cell lines, stomach cancer cell line and prostate cancer cell line, with IC₅₀ less than 10 μM, indicating its potential activity against cancer cells.     

Genetic variation of the Chinese longsnout catfish Leiocassis longirostris in the Yangtze River revealed using mitochondrial DNA cytochrome b sequences

The mitochondrial DNA cytochrome b of 132 Leiocassis longirostris collected from 12 localities in the upper to lower reaches of the Yangtze River were amplified and partially sequenced using the PCR technique. The results showed that 27 nucleotide sites were variable along 817 bp length of homologous sequence (3.3%), base substitutions happened mostly at the third codon position. A total of 22 haplotypes were identified, which were characterized with moderate haplotype diversity (h = 0.5417 ± 0.0519), but low nucleotide diversity (π = 0.0019 ± 0.0012). Median-joining network analysis revealed star-shaped patterns with one common central haplotype (H3), whereas mismatch distribution analysis found that the Chinese longsnout catfish fitted a smooth unimodal distribution, which suggested that this species underwent population expansion following bottlenecks and/or they originated from a small number of founding individuals. The time that the total population of Chinese longsnout catfish in the Yangtze River expanded was estimated 169,000–337,000 years before present. The analysis of molecular variance (AMOVA) and phylogenetic reconstructions did not detect significant geographic structure between different river sections, especially between above and below the Gezhouba Dam and the Three Gorges Dam, which suggested that these recently developed dams might have not significantly resulted in population genetic differentiation in the Chinese longsnout catfish.