芋螺毒素的环化研究

芋螺毒素生物活性强,对药物作用靶点的选择性高,在治疗神经性疼痛等疾病方面具有很好的开发前景。然而天然芋螺毒素容易受到体内蛋白酶的水解。将其进行环化,可增加它们在人体内的稳定性,从而具有口服活性。本文以α-芋螺毒素Vc1.1和MII环化后的稳定性与活性变化、环化的合成方法为例,对芋螺毒素的环化研究进展进行综述,说明环化的芋螺毒素将有可能在治疗神经性疼痛等疾病中扮演一个重要的角色。     

毕赤酵母高效电转化条件的研究

目的建立毕赤酵母的高效电转化方法,为利用毕赤酵母重组表达海洋药物功能基因提供方法基础。方法采用电穿孔的方法把外源DNA转化进毕赤酵母基因组中。通过对毕赤酵母生长状态,电击条件等影响因素进行探索。结果当采用对数生长中期的菌体制备感受态细胞、电压为1.5kV、质粒浓度为0.15g.L-1和0.2cm电转化杯时,转化效率达到最大值,为每微克质粒DNA 36个转化子。经抽样PCR鉴定所得到的转化子均为阳性克隆。结论通过对电转化各种条件的优化,获得了高效电转化技术方法。     

南药益智果实的总RNA提取方法比较研究

目的筛选从南药益智不同发育时期的果实中提取出高质量RNA的试剂盒。方法以南药益智果实为研究对象,比较了3种Mag Zol TM Reagent,Takara RNAiso plus和RNAprep Pure Plant Kit试剂盒提取方法,利用琼脂糖凝胶电泳、Nano Drop TM 2000分光光度计和Aligent 2100生物分析仪对3种方法提取的总RNA进行质量检测。结果采用RNAprep Pure Plant Kit试剂盒从南药益智不同发育时期果实中提取的RNA电泳条带清晰、RNA完整性好,其纯度(OD260/280)分别为2.04、2.08和2.06;浓度分别为68.12 ng/μl、140.20 ng/μl和181.51 ng/μl。结论试剂盒RNAprep Pure Plant Kit可有效地从富含多酚益智果实中提取高质量RNA,为南药益智的高通量转录组测序等分子生物学研究奠定基础。     

海葵多肽神经毒素的研究新进展

海葵具有用于捕食和防御功能的触手刺细胞,其分泌的毒液主要包含蛋白和多肽类毒素,其中多肽神经毒素具有序列多样,结构稳定,靶点特异等特点。目前,已经从海葵中分离并鉴定了100余种多肽神经毒素,以电压门控钠离子通道（voltage-gated sodium channels,VGSCs）、电压门控钾离子通道（voltage-gated potassium channels,VGPCs）、酸敏感离子通道（acid-sensing ion channels,ASICs）等为主要作用靶点,它们已经成为研究特定离子通道的结构和功能的重要工具。本论文主要综述迄今为止从海葵中分离得到的海葵多肽神经毒素的类型、结构特征、药理活性以及临床应用。     

基于SRAP技术分析海南产南药益智的遗传多样性＊

目的 海南产南药益智为道地药材，为探究海南岛不同产地的南药益智资源的遗传多样性。方法 通过对144对SRAP引物进行筛选和优化，获得12对较好的引物，用以对海南岛内15个不同产地的90份样品进行遗传多样性分析。结果 SRAP-PCR扩增结果获得总条带数（TNB）为92，多态性条带数（NBP）为82，平均多态率（PPB）为89.1%。再以最佳引物f9-r10对90份益智样品进行SRAP-PCR分析，统计共获得407个条带，多态性条带数为407个，多态性比率为100%。聚类结果分析表明，不同产地益智的遗传相似系数在0.67-1.0之间，在遗传相似系数0.67处90份益智样品被分为Ⅰ和Ⅱ两大类，在遗传相似系数0.77处可分为6个亚类。结论 不同产地的南药益智遗传多样性与产地具有一定的相关性，来自五指山的益智遗传多样性最为丰富，为种质资源保护提供理论基础。     

银杏平颤方对帕金森病模型小鼠多巴胺神经元丢失及其凋亡的影响

BACKGROUND: Current pharmacological research has administrated that Ginkgo biloba and its extracts can eliminate loss and apoptosis of dopamine neurons by antioxidation and nerve growth factor activation. OBJECTIVE:To confirm the effects of Ginkgo biloba Pingchan Recipe on loss and apoptosis of dopamine neurons as well as proliferation of neuroblastoma derived cell lines SH-SY5Y in mice with Parkinson’s disease. METHODS: (1) C57BL mice were randomly divided into three groups: 20 mice were modeled into Parkinson’s disease by intraperitoneal injection of MPTP for 6 weeks, and at 30 minutes before each intraperitoneal injection, mice in model group were given gavage of normal saline, mice in treatment group given Ginkgo biloba Pingchan Recipe. The other 10 mice received no any interventions as normal group. The loss and apoptosis of dopamine neurons were observed by immunohistochemistry at 15 and 30 days after modeling. (2) In vitro cultured SH-SY5Y cell lines were treated with Ginkgo biloba Pingchan Recipe. Subsequently, the cell proliferation and apoptosis were detected using MTT assay and flow cytometry, respectively; PARP and PTEN mRNA and protein levels were measured through real-time PCR and western blot assay. RESULTS AND CONCLUSION: (1) Compared with the model group, the number of tyrosine hydroxylase-positive neurons at the substantia nigra compacta of midbrain was significantly increased in a time-dependant manner in the normal and treatment groups (P < 0.05). (2) The apoptosis rate and mRNA and protein levels of PARP and PTEN in the treatment group were lower than those in the model group. These results suggest that Ginkgo biloba Pingchan Recipe can partly inhibit midbrain neuron apoptosis that may be by decreasing PTEN level. As a result, it becomes possible to prevent and treat Parkinson’s disease via protecting dopamine neurons from loss. 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

海南白沙县青松乡人工栽培益智种质资源的调查与分析

目的:对海南白沙县青松乡栽培的益智种质资源进行调查,对3个主要益智种质资源的外观形态和次生代谢产物含量进行评价,为益智优质种质资源选育提供基础。方法:测量益智不同种质资源叶片的长度和宽度,称量益智不同种质资源的果实粒重,用LC-MSMS分析不同种质资源果实中次生代谢产物的含量。结果:不同益智质资源的叶片长宽度、果实粒重和次生代谢产物含量间具有明显差异,以圆果益智的质量最佳。结论:益智不同种质资源的外观形态和次生代谢产物含量差异较大,圆果益智为优质品种,建议推广种植。     

苯酚-硫酸法测定中药海马总多糖的含量的研究

目的建立中药海马总多糖含量测定方法。方法以葡萄糖标准品为对照品,采用苯酚-硫酸法测定海马冷浸多糖、热提多糖、碱提多糖中总多糖的含量测定。结果三种多糖总多糖含量差异显著,其中碱提多糖含量最高,其次热提多糖,最后冷浸多糖。结论测定方法简单准确快速。