AFLP analysis of genetic diversity and relationship among some Chinese domestic ducks and wild ducks

The amplified fragment length polymorphic(AFLP)technique was used to analyze the genome DNA polymorphism among 8 breeds of domestic ducks and 2 species of wild ducks.Nine of the 17 selected primers pairs gave reproducible polymorphic DNA amplification bands.The amplified bands ranged from 44 to 83 per primer pair.Of the 513 AFLP markers obtained.498 were polymorphic.The proportion of polymorphic loci was 97.1%.The genetic distance(D)and similarity coefficients(GS)were calculated based on the polymorphic data.Between domestic ducks D ranged from 0.331 to 0.589,while between domestic ducks and the wild ducks,it ranged from 0.298 to 0.520(vs.Anas Platyrhynchos)and from 0.316 to 0.522(vs.A.Poecilorhyncha),respectively.The variance analysis showed no significant difference between the two groups of data,which indicated that both mallard and spot-billed ducks made contributions to domestic duck evolution.A dendrogram was constructed according to the D value.     

A “GC-rich” method for mammalian gene expression: A dominant role of non-coding DNA GC content in regulation of mammalian gene expression

High mammalian gene expression was obtained for more than twenty different proteins in different cell types by just a few laboratory scale stable gene transfections for each protein. The stable expression vectors were constructed by inserting a naturally-occurring 1.006 kb or a synthetic 0.733 kb DNA fragment (including intron) of extremely GC-rich at the 5′ or/and 3′ flanking regions of these protein genes or their gene promoters. This experiment is the first experimental evidence showing that a non-coding extremely GC-rich DNA fragment is a super “chromatin opening element” and plays an important role in mammalian gene expression. This experiment has further indicated that chromatin-based regulation of mammalian gene expression is at least partially embedded in DNA primary structure, namely DNA GC-content.     

重组人维甲酸X受体α的克隆与表达及其与甲状腺受体的结合

维甲酸X受体α(Retinoid X receptor-α,RXR-α)属于核受体家族,在调节基因转录及信号转导方面发挥着重要的作用。为了深入研究RXR-α的生物学作用,文章利用RT-PCR技术扩增了人RXR-α基因,并将其克隆入原核表达载体pQE-30Xa,转化大肠杆菌M15[PREP4],异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达,SDS-PAGE可见与预期大小相符的蛋白条带,Western blotting证实该条带为重组人RXR-α蛋白,并用Ni2-NTA柱进行亲和层析纯化。免疫共沉淀结果显示其具有与TRβ1结合的能力。电泳迁移率改变实验(EMSA)显示RXR-α与TRβ1形成的杂二聚体具有与DNA结合的能力。结果显示,文章已成功建立了重组人RXR-α的大肠杆菌表达系统,表达产物具有很好的生物学活性。     

中心体蛋白Cenexin在大鼠精子发生过程中的表达特征

中心体蛋白Cenexin是成熟中心粒的唯一标志分子。为阐明中心粒在大鼠精子发生中的成熟以及功能,我们首先通过RT－PCR技术从大鼠睾丸组织中扩增出了Cenexin cDNA片段,原核表达重组蛋白后,用其免疫小鼠制备了高滴度的抗Cenexin的多克隆抗体,然后利用免疫荧光染色、Western Blot和半定量RT－PCR方法,研究了大鼠精子发生过程中Cenexin蛋白和基因的表达特征。结果显示Cenexin mRNA水平在精原细胞和精母细胞中较高,随后表达水平下降,而蛋白质分子在精原细胞到精子细胞中都定位于细胞的一个中心粒上,表示有成熟中心粒的存在,在长形精子细胞中该蛋白位于鞭毛的基体部。附睾的绝大多数成熟精子中Cenexin免疫染色消失。中心体蛋白Cenexin在精子变态期的表达变化可能与精子鞭毛形成的起始有关。     

芦笋皮层组织培养植株再生

报道了芦笋皮层组织培养再生植株的方法。表明：皮层在ＭＳ＋１ｍｇ／Ｌ２,４—Ｄ＋２ｍｇ／Ｌ６—ＢＡ培养基上形成无色或淡绿色疏松型愈伤组织,而ＭＳ＋２ｍｇ／Ｌ６—ＢＡ＋０．１ｍｇ／ＬＩＡＡ培养基则对芽的分化有利;试管苗的生很壮苗以无激素的ＭＳ为最佳培养基。此外,本试验还研究了不同碳源、不同激素组合对芦笋皮层离体培养再生植株的影响。     

The mechanism for the effect of selenium supplementation on immunity

Lipid peroxide (LPO) in lymphocytes from mice was evaluated by measuring substances reactive to thiobarbituric acid (TBA). The product resulting from the reaction of TBA with lymphocytes was extracted with n-butyl and fluorescence intensity was determined. The degree of lipid peroxidation, expressed as fluorescence intensity f547, was assessed for stimulation of lymphocytes with concanavalin A (Con A), and was related to lymphocyte proliferation in response to Con A if Se was administered. The lymphocyte proliferation was determined by [³H]thymidine incorporation, expressed as cpm. The effect of superoxide dismutase (SOD), added to cell culture on lymphocyte proliferation was also evaluated. It was found that LPO in lymphocytes before Con A stimulation was significantly less than that after stimulation (p<0.001), and that SOD promoted lymphocyte proliferation dose dependently. The addition of Na₂SeO₃ to lymphocyte culture or supplementation in drinking water to mice decreased the produced LPO in lymphocyte in response to Con A. In the presence of Se, there is an inverse correlation between the levels of LPO in lymphocyte and the stimulated proliferation (r=−0.8902,r=−0.9439). In conclusion, active oxygen species scavenging was proposed as one of the mechanisms for Se to promote immunity.     

青藏公路沿线白昼交通运输等人类活动对藏羚羊迁徙的影响

To study effects of traffic during daytime and other human activities, we conducted surveys on the migration of Tibetan antelope Pantholops hodgsoni along the Qinghai-Tibet highway between Kunlun Mountain Pass to Wudaoliang,using line transects (LT) from June to September in 2001 and 2002, Data were collected for 64 days with 17 days in 2001, and 47days in 2002. The Stationary Observation Method (SOM) was also adopted for the investigation of status of the disturbance from traffic when antelopes passing the highway and the traffic situation from August 6 to 28 (6 : 30-19: 00), 4 to 20 (6: 00- 18: 00) and from December 21 to 29 (9：00- 19: 00) in 2002. The results showed that,the period of migration in 2002 was more prolonged than in 2001, and it took at least 45 min for each group to cross the highway, with success ratio of only 30.2 %, and the traffic flow of summer daytime was 76/h, while the winter‘s traffic flow was 29/h. Highway and rail traffic should be managed during daylight hours from June to August, to allow for the migration of the antelopes. Also conservation education and a further survey should be emphasized for the protection on the species [Acta Zoologica Sinica 50 (4) : 669-674, 2004].     

3D-QSAR studies of checkpoint kinase 1 inhibitors based on molecular docking and CoMFA

Three-dimensional quantitative structure–activity relationship (3D-QSAR) studies were performed on a series of substituted 1,4-dihydroindeno[1,2-c]pyrazoles inhibitors, using molecular docking and comparative molecular field analysis (CoMFA). The docking results from GOLD 3.0.1 provide a reliable conformational alignment scheme for the 3D-QSAR model. Based on the docking conformations and alignments, highly predictive CoMFA model was built with cross-validated q ² value of 0.534 and non-cross-validated partial least-squares analysis with the optimum components of six showed a conventional r ² value of 0.911. The predictive ability of this model was validated by the testing set with a conventional r ² value of 0.812. Based on the docking and CoMFA, we have identified some key features of the 1,4-dihydroindeno[1,2-c]pyrazoles derivatives that are responsible for checkpoint kinase 1 inhibitory activity. The analyses may be used to design more potent 1,4-dihydroindeno[1,2-c]pyrazoles derivatives and predict their activity prior to synthesis.     

脂肪酶在微乳液和微乳液凝胶中催化辛酸辛醇的酯化反应

脂肪酶在合成反应中具有很高的区域选择性和立体选择性 ,已广泛用于食品工业和药物工业[1,2 ] ,在有机介质中的脂肪酶催化反应已有较多研究[3 ,4 ] 。微乳液一般由表面活性剂、助表面活性剂、油和水等组份组成 ,它是一种热力学稳定、光学透明、宏观均匀而微观不均匀的体系 ,能提供酶催化所需要的巨大油 /水界面[5] 。而将脂肪酶增溶于油包水(Ｗ /Ｏ)微乳液中的纳米级“水池”中 ,可使酶以分子水平分散[6] ,图 1(ａ) ,从而可用来模拟细胞微环境中的反应。油包水微乳液中的酶可通过加入明胶而制成固定化酶 ,含明胶的微乳液凝胶 (ＭＢＧｓ)最早…     

Enhanced intrinsic migration of aggressive breast cancer cells by inhibition of Rac1 GTPase

Rac GTPases are known to play a crucial role in regulating cytoskeletal changes necessary for cell migration. Migration has been shown to be positively regulated by Rac in most cell types. However, there is also a large body of conflicting evidence in some other cell types with respect to the role of Rac in migration, suggesting that Rac GTPases regulate cell migration in a cell type-dependent manner. In the present study, we have characterized the effects of Rac1 GTPase inhibition on the migratory abilities of a number of breast cancer cell lines with differential degrees of tumorigenic and metastatic potentials. We show that Rac1 inhibition in non-metastatic (MCF-7, T-47D) or moderately metastatic (Hs578T) cell lines results in inhibition of migration, whereas in highly metastatic cell lines (MDA-MB-435, MDA-MB-231, and C3L5) Rac1 inhibition results in stimulation of migration. This stimulation of migration following Rac1 inhibition is also accompanied by the enhanced RhoA activity, suggesting a possible existence of a dominating role of RhoA over Rac1 in regulating intrinsic migration of the highly metastatic breast cancer cells.