不同预处理对小鼠粪样菌群高通量测序分析结果的影响

应用高通量测序技术比较不同预处理对小鼠粪样菌群结构的影响,以期为后续相关研究提供参考依据。采集昆明小鼠新鲜粪样,分为原始粪样组、生理盐水处理组和PBS处理组,提取粪样DNA,采用Illumina MiSeq平台进行测序对3组样本的16S rRNA V3~V4区基因文库进行生物信息学分析。结果显示,3组样本拥有共同的OTUs 188个,不同预处理对粪样微生物多样性和丰度产生影响,生理盐水处理组、PBS处理组和原始粪样组分别检出14、11、12个门,3组样本共有门11个;分别检出24、21、21个纲,3组样本共有纲20个;分别检出62、55、59个科,3组样本共有科26个;分别检出147、126、137个属,3组样本共有属117个。总体来看,生理盐水处理组微生物多样性和丰度相对最高,原始粪样组次之,PBS处理组最低。粪样经生理盐水处理后,能在一定程度上提高肠道菌群检测的准确性。     

金银花花形基因的发掘及生物信息学分析

金银花作为我国重要的中药材,具有消炎、抗菌、抗病毒、抗氧化、防癌等多种功效。随着金银花市场供需矛盾日益加剧,通过分子标记辅助选择育种方法来培育高产优质品种势在必行。通过NCBI的Blast工具扫描金银花蛋白组数据发掘花形候选基因,并执行候选基因的亲缘关系分析、结构域分析、表达模式分析、理化性质分析、蛋白质结构预测等一系列生物信息学分析。依据拟南芥调控花形的ABE类基因,通过NCBI-Blast工具扫描金银花氨基酸序列,筛选出包含MADS结构域的8个调控花形的金银花候选基因。经LjaFGD表达模式分析发现,金银花的花中GWHGAAZE016592和GWHGAAZE014905表达量显著高于其他部位,可能正向调控金银花花形。GWHGAAZE014905是一个包含MADS结构域的调控花器官发育的B类基因;GWHGAAZE016592是AP3同源基因。生物信息学分析发现,GWHGAAZE016592和GWHGAAZE014905均是稳定的亲水蛋白,属于非分泌蛋白,包括Motif1、Motif3、Motif4、Motif2、Motif6和Motif5,蛋白质三级结构模板为6byy.2.A和4ox0.2.C。GWHGAAZE014905被定位到细胞核上,而GWHGAAZE016592被定位到叶绿体上,且包含1个位于151~173 bp的跨膜螺旋区域,属于膜蛋白。研究结果为分子标记辅助选择方式培育道地高产优质金银花品种提供了基因资源和分子标记。     

Dampening HOTAIR sensitizes the gastric cancer cells to oxaliplatin through miR-195-5p and ABCG2 pathway

Long non-coding RNAs (lncRNA) have an extensive role in the progression and chemoresistance of gastric cancer (GC). Deeply study the regulatory role of lncRNAs could provide potential therapeutic targets. The aim of this study is to explore the regulatory role of HOTAIR in the progression and oxaliplatin resistance of GC. The expression of HOTAIR in GC and cell lines were detected by using qRT-PCR. Cell proliferation and apoptosis were analysed by CCK-8, EdU incorporation and flow cytometry. Luciferase reporter assay was used to identify the interaction between HOTAIR and ABCG2 (ATP-binding cassette (ABC) superfamily G member 2, ABCG2) via miR-195-5p. The regulatory functions were verified by using molecular biology experiments. HOTAIR was significantly overexpressed in GC and associated with poor prognosis. Knock-down of HOTAIR inhibited the GC cells proliferation and oxaliplatin resistance, while overexpression of HOTAIR showed opposite functions. Further studies found that HOTAIR acted as a competing endogenous RNA (ceRNA) to absorb miR-195-5p and elevated the expression of ABCG2, which leads to resistance of GC cells to oxaliplatin. Taken together, our findings demonstrated that HOTAIR regulates ABCG2 induced resistance of GC to oxaliplatin through miR-195-5p signalling and illustrate the great potential of developing new therapeutic targets for GC patients.     

Systematics of Mukdenia and Oresitrophe (Saxifragaceae): Insights from genome skimming data

Oresitrophe and Mukdenia (Saxifragaceae) are epilithic sister genera used in traditional Chinese medicine. The taxonomy of Mukdenia, especially of M. acanthifolia, has been controversial. To address this, we produced plastid and mitochondrial data using genome skimming for Mukdenia acanthifolia and Mukdenia rossii, including three individuals of each species. We assembled complete plastomes, mitochondrial CDS and nuclear ribosomal ETS/ITS sequences using these data. Comparative analysis shows that the plastomes of Mukdenia and Oresitrophe are relatively conservative in terms of genome size, structure, gene content, RNA editing sites and codon usage. Five plastid regions that represent hotspots of change (trnH-psbA, psbC-trnS, trnM-atpE, petA-psbJ and ccsA-ndhD) are identified within Mukdenia, and six regions (trnH-psbA, petN-psbM, trnM-atpE, rps16-trnQ, ycf1 and ndhF) contain a higher number of species-specific parsimony-informative sites that may serve as potential DNA barcodes for species identification. To infer phylogenetic relationships between Mukdenia and Oresitrophe, we combined our data with published data based on three different datasets. The monophyly of each species (Oresitrophe rupifraga, M. acanthifolia and M. rossii) and the inferred topology ((M. rossii, M. acanthifolia), O. rupifraga) are well supported in trees reconstructed using the complete plastome sequences, but M. acanthifolia and M. rossii did not form a separate clade in the trees based on ETS + ITS data, while the mitochondrial CDS trees are not well-resolved. We found low recovery of genes in the Angiosperms353 target enrichment panel from our unenriched genome skimming data. Hybridization or incomplete lineage sorting may be the cause of discordance between trees reconstructed from organellar and nuclear data. Considering its morphological distinctiveness and our molecular phylogenetic results, we strongly recommend that M. acanthifolia be treated as a distinct species.     

水稻蜡质基因5＇上游区缺失对基因表达的影响

为研究水稻蜡质基因（ｗａｘｙ）５＇上游调控区中存在的顺式作用元件,我们将水稻ｗａｘｙ基因翻译起始声、（ＡＴＧ）５＇上游３．４ｋｂ（－２１１８～＋１２９１ｂＰ）片段经外切核酸酶ＥｘｏⅢ部分酶解,得到一系列５＇端缺失的片段。将这些缺失片段分别与ｇｕｓ基因编码区连接,构建成融合质粒,经ＰＥＧ介导引入水稻原生质体,２６℃培养４８ｈ后,定量测定ＧＵＳ酶活力,并以同时导入的由３５Ｓ启动子指导的荧光素酶（ＬＵＣ）基因表达的酶活力作为内对照。结果表明,ＧＵＳ酶活性随５’上游调控区长度的减少而逐渐减弱。由─８６１ｂｐ缺失至─６４０ｂＰ时,ｇｕｓ基因表达水平有较明显的降低,推测在该区域中可能存在一个顺式作用元件区。     

脏器微血管对荧光素钠通透性的实验方法

大鼠颈动脉注射１％ＦｌＮａ,荧光显微镜下活体观察肠系膜微血管血流状态及ＦｌＮａ的渗出情况,并在不同时间点经股动脉采血,测定血浆内ＦｌＮａ浓度随时间的变化,利用组织匀浆测定不同脏器中ＦｌＮａ的分布,再辅以冰冻切片进行观察。活体观察发现,ＦｌＮａ注入体内后,经微血管迅速向周围组织渗出,最后汇集于淋巴管,血浆及组织匀浆ＦｌＮａ浓度的测定表明,ＦｌＮａ浓度随时间的变化呈指数衰减,各脏器ＦｌＮａ的分布极不相同。冰冻切片也显示了同样的分布差别。这些结果表明,我们所建立的方法可直观、定量地反映ＦｌＮａ在微血管的通透情况。     

人体超微弱发光图像中的信号检验

用近期研制的具有单光子探测能力的超高灵敏度成像系统获得了人体体表生物超微弱发光的强度数据。为分析图像中的信号检验,二项分布被用于人手不同时间累积发光图像中的信号显著性检验,证实人手存在超微弱生物发光,手指的发光强度在３００～５５０ｐｈｏｔｏｎｓ／ｓ范围内,全手的发光强度在８５０～１２００ｐｈｏｔｏｎｓ／ｓ范围内。     

Reversion of malignancy in human gastric cancer MKN—45 cells through the transfection of transforming growth factor—β type Ⅱ receptor gene

Human gastric cancer MKN-45 cells which are resistant to TGF-β growth inhibition and possess TGF-β type I and type Ⅲ receptors,but not type Ⅱ receptors,have been used as a model system to reconstitute these cancer cells with TGF-β RII cDNA.The results of these experiments indicated that the reexpression of TGF-β RII gene in MKN-45 cells can restore their sensitivity to TGF-β growth inhibition,decrease their growth rate,reduce their cloning efficiency in soft agar and tumorigenicity in nude mice in stable transfectants,in comparison with their control MKN-45 cells.Among different RII transfectants,their difference in the changes of these parameters,as a result of the regain of autocrine negative growth control by TGF-β,is roughly proportional to their level of expression of transfected RII mRNA.From these data,it is concluded that the inactivation of TGF-β RII gene is related to the escape of growth control by TGF-β in MKN-45 cells.The importance of the study of the interplay of TGF-β and its receptor system in the negative growth control of gastric cancer,and possibly also of other cancers,is discussed.     

高压静电场处理沙棘插条生根状况的初步研究

用高压静电场处理沙棘插条研究生根状况,方法简便易于处理,该装置适于进行大规模的处理,从目前研究表明：高压静电场对沙棘插条有正刺激效应,并以电场强度为３．９ｋｖ／ｃｍ,时间为４０分钟处理最佳。