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.     

Using Molecular Dynamics to Predict Factors Affecting Binding Strength and Magnetic Relaxivity of MRI Contrast Agents

We demonstrate the use of molecular dynamics and molecular mechanics methods to calculate properties and behavior of metal-chelate complexes that can be used as MRI contrast agents. Static and dynamic properties of several known agents were calculated and compared with experiment. We calculated the static properties such as the q-values (number of inner shell waters) and binding distances of chelate atoms to the metal ion for a set of chelates with known X-ray structure. The dynamic flexibility of the chelate arms was also calculated. These computations were extended to a series of exploratory chelate structures in order to estimate their potential as MRI contrast agents. We have also calculated for the first time the NMR relaxivity of an MRI contrast agent using a long (5 nsec) molecular dynamics simulation. Our predictions are promising enough that the method should prove useful for evaluating novel candidate compounds before they are synthesized. One novel static property, the projected area of chelate atoms onto a virtual surface centered on the metal ion (gnomonic projection), was found to give an effective measure of how well the chelate atoms use the free space around the metal ion.     

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。     

HCV5＇端非编码区cDNA的体外转录

采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）从广东省一例慢性丙型肝炎病人血清中获得丙型肝炎病毒（ＨＣＶ）５＇端非编码区（５＇ＮＣＲ）３０２ｂｐ的ｃＤＮＡ片段,经补齐和提纯后插入ｐＵＣ１９质粒,获得的重组体ｐＵＮ进行序列测定。将ｐＵＮ的目的基因亚克隆进体外转录载体ｐＳＰＯＲＴＩ多克隆位点的ＥｏｏＲＩ和ＰｓｔＩ切点之间,所得重组体ｐＳＮ线性化后由Ｔ＿７ＲＮＡ多聚酶及ＳＰ６ＲＮＡ多聚酶引导体外转录反应,产物经凝胶电泳及特异引物ＲＴ－ＰＣＲ,证实ＳＰ６引导的是正义ＲＮＡ,Ｔ７合成的是反义ＲＮＡ,其大小分别力４２９ｂｐ和３６２ｂｐ。并证实所得ＲＮＡ力ＨＣＶ５＇ＮＣＲｃＤＮＡ转录而来。获得的ＨＣＶ５＇ＮＣＲｃＤＮＡ和ＲＮＡ在常规逆转录和ＰＣＲ步骤中用于设立有效的模板对照,对消除假用性及评估试剂有重要意义。同时,ＨＣＶ５＇ＮＣＲ体外转录载体的构建可用于制各ＲＮＡ探针和反义ＲＮＡ,改进后还可作为定量ＰＣＲ的竞争性模板。     

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。     

用细胞色素C法检测非真空电子束辐射的间接作用

本文提出了一种检测非真空电束辐射的间接作用的一种方法,根据氧化型和还原型的细胞色素Ｃ的吸收光谱不同,从而测量非真空电子束作用对应的辐射分解的自由基总量。     

Expression of betaine aldehyde dehydrogenase gene and salinity tolerance in rice transgenic plants

Betaine as one of osmolytes plays an important role in osmoregulation of most high plants. Betaine aldehyde dehydrogenase C BADH) is the second enzyme involved in betaine biosynthesis. The BADH gene from a halophite, Atriplex hortensis, was transformed into rice cultivars by bombarment method. Totally 192 transgenic rice plants were obtained and most of them had higher salt tolerance than controls. Among transgenic plants transplanted in the saline pool containing 0.5% NaCl in a greenhouse, 22 survived, 13 of which set seeds, and the frequency of seed setting was very low, only 10% . But the controls could not grow under the same condition. The results of BADH ac-tivity assay and Northern blot showed that the BADH gene was integrated into chromosomes of transgenic plants and expressed.     

Binding characterization of the iron transport receptor from the outer membrane of Escherichia coli (FepA): differentiation between FepA and FecA

The dissociation constants for the binding of ferric enterobactin with FepA and FecA are quantitated with displacement experiments. It is found that K _d for FepA is 12 times lower than the one for FecA. This indicates that FepA is an high-affinity receptor while FecA binds ferric enterobactin with a lower affinity. Monoclonal antibodies specific for binding epitopes of FepA inhibit the binding of ferric enterobactin with purified FepA. These same antibodies do not inhibit the binding of ferric enterobactin with purified FecA. This indicates that the binding epitopes in FecA and FepA are different.