香橼优良单株半同胞家系子代抗寒性研究

在南京六合地区对3年生24个香橼优良单株半同胞家系子代进行抗寒性测定研究,结果表明家系子代间抗寒性存在着显著的差异。田间观测-11℃自然低温下春江4#和香橼4#的抗寒性最强,受冻株率分别为86.67%和100%,平均受冻级别均为Ⅰ级,耐寒指数均为2.00;新世纪8#、春江7#和新世纪2#的抗寒性最差,受冻株率均为100%,平均受冻级别均达Ⅲ级,耐寒指数均大于3.00。室内抗寒力测定供试家系子代的半致死温度在-14.84℃～-7.05℃之间。春江4#和香橼4#的抗寒力最强,半致死温度分别为-14.84℃和-13.94℃;春江7#和新世纪2#的抗寒力最差,半致死温度分别为-7.91℃和-7.05℃。抗寒性评估结果一致。根据半致死温度,结合田间抗寒性观测,供试香橼家系子代抗寒性评估排序为春江4#>香橼4#>春江6#>春江5#>新世纪9#>新世纪10#>新世纪3#>香橼1#>新世纪7#>香橼7#>新世纪8#>香橼6#>香橼5#>春江3#>新世纪4#>新世纪6#>香橼2#>春江8#>新世纪5#>春江1#>新世纪1#>春江2#>春江7#>新世纪2#。     

Antiproliferative activity of Saponaria vaccaria constituents and related compounds

Total methanolic extracts of Saponaria vaccaria seed derived from several varieties, as well as various purified components obtained through successive chromatographic separations of total extracts were evaluated for their growth inhibitory activity in WiDr (colon), MDA-MB-231 (breast), NCI-417 (lung) and PC-3 (prostate) human cancer cells as well as the non-tumorigenic fibroblast BJ (CRL-2522) cell line using MTT colorimetric assay. Purified bisdesmosidic saponins segetoside H and I were further examined using microscopy and apoptosis assays. Bisdesmosidic saponins exhibited dose-dependent growth inhibitory and selective apoptosis-inducing activity. Growth inhibitory effects were particularly strong in a breast (MDA-MB-231) and a prostate (PC-3) cancer cell line. Total extracts exhibited a different preference being most active against a colon cancer cell line (WiDr). In a comparison of varieties, all of the total seed extracts exhibited similar dose-dependent activities, but with some variation in potency. Monodesmosidic saponins vaccarosides A and B, phenolic vaccarin, and cyclopeptide segetalin A, co-occurring seed substituents, did not exhibit activity. The non-tumorigenic fibroblast cell line BJ (CRL 2522) was growth inhibited but did not undergo apoptosis when treated with bisdesmosidic saponins at low micromolar concentrations. Saponin-rich extracts from Kochia scoparia seed and Chenopodium quinoa were also evaluated alongside Saponaria saponins but did not exhibit activity. Closely related Quillaja saponins exhibited activity but were less potent.     

蒲公英内生真菌PG23抗癌活性的初步研究

研究药用植物蒲公英内生真菌PG23多糖对肺癌A549细胞的抑制作用。蒲公英内生真菌PG23通过培养,乙醇沉淀获得粗多糖、苯酚-硫酸法测定多糖含量,采用MTT法测定PG23多糖对肺癌A549细胞的抑制率,并计算半抑制浓度(IC50)。结果表明,PG23发酵液中多糖得率为2.754%,多糖含量为26.82%;对A549肺癌细胞的抑制率以第1天为最高;抑制率不与多糖浓度成比例,最佳作用浓度为14.42μg/mL,抑制率为87.63%(P<0.01),IC50为1.073mg/mL(24h)。PG23发酵液多糖对肺癌A549细胞具有较强的抑制能力。     

3种半数致死浓度计算方法之比较

通过对寇氏法、概率单位法及线性回归法等3种半数致死浓度(50%lethal concentration,LC50)计算方法的阐述,比较了其基本公式与操作方法等。结果表明不同时间的半数致死浓度均依线性回归法、寇氏法、概率单位法的顺序逐步升高,但差异性不显著(P>0.05)。在操作上,概率单位法和线性回归较寇氏法更容易掌握。     

Practical capability of a DNA pool‐based genome‐wide association study using BovineSNP50 array in a cattle population

Genome‐wide association mapping for complex traits in cattle populations is a powerful, but expensive, selection tool. The DNA pooling technique can potentially reduce the cost of genome‐wide association studies. However, in DNA pooling design, the additional variance generated by pooling‐specific errors must be taken into account. Therefore, this study aimed to investigate factors such as: (i) the accuracy of allele frequency estimation; (ii) the magnitude of errors in pooling construction and in the array; and (iii) the effect of the number of replicate arrays on P‐values estimated by a genome‐wide association study. Results showed that the Illumina correction method is the most effective method to correct the allele frequency estimation; pooling errors, especially array variance, should be taken into account in DNA pooling design; and the risk of a type I error can be reduced by using at least two replicate arrays. These results indicate the practical capability and cost‐effectiveness of pool‐based genome‐wide association studies using the BovineSNP50 array in a cattle population.     

莲雾不同种的低温半致死温度及抗冷适应性

以叶片和成熟枝条为试材,应用电导法测定莲雾在不同低温下细胞膜透性的变化,配合Logistic方程求出低温半致死温度(LT50),评价6个莲雾种在不同降温时期的抗冷性,及同一时期不同部位的抗冷性,并结合露地栽培越冬表现进行验证。结果表明,在自然降温过程中,6个莲雾种的低温半致死温度均随气温的下降而不断降低,但下降幅度因种而异,1.69～2.97℃不等。6个种抗冷性由强到弱排序依次为水蒲桃>本地种>粉红种>紫红种>青色种>印度红。2 a生枝条的抗冷力最强,明显比叶片耐冷。冬季越冬表现与1月半致死温度测定结果基本一致,表明半致死温度可作为莲雾抗冷性评价的一个可靠指标。长时间低温加上霜冻,使露地栽培本地种、粉红种、紫红种的叶片和部分嫩梢受冷害,而印度红、青色种不能越冬成活。     

披针叶黄华碱对小鼠半数致死量的测定

目的：以小鼠为试验动物,对披针叶黄华碱的毒性进行研究,测定其生物碱组分的半数致死量.方法：用体积分数为95％的乙醇结合超声波对披针叶黄华植物进行提取,以小鼠为试验动物,研究拨针叶黄华碱氯仿萃取组分的毒性.结果：通过试验测得披针叶黄华碱萃取组分的半数致死量为127.04 mg/kg,95％可信限为115.41-139.72 mg/kg.结论：试验结果与报道数据差异较大,可能与试验方法、萃取药液的实际浓度等因素有关.     

乙型脑炎病毒荧光定量PCR的建立及对病毒体外复制动态的研究

根据乙型脑炎病毒(JEV)NS3基因保守序列设计并合成一对引物,以构建的重组质粒作为阳性标准品,建立了一种敏感、特异、重复性好的快速检测JEV的SYBR Green I荧光定量PCR方法.该方法在1.92×108～1.92×101拷贝/μL范围内具有良好的线性关系,可检测到初始模板中19拷贝/μL的病毒核酸,敏感性是常规RT-PCR方法的10倍;检测时间缩短了50％,该方法不与其它猪源病毒发生交叉反应;在重复性试验中,批间、批内变异系数均小于1.2％.应用本方法对JEV在BHK-21细胞上繁殖滴度进行定量检测,绘制JEV在BHK-21细胞上的一步生长曲线,并与TCID50方法绘制的复制动态曲线进行比较.结果显示,两种方法测定的JEV在BHK-21细胞上的复制动态具有一定的平行关系,对于制备灭活疫苗而言,荧光定量PCR方法更快速和敏感,所测得的抗原含量更精确,有利于企业机动灵活地安排生产.     

Insights into the Toxicological Properties of a Low Molecular Weight Fraction from Zoanthus sociatus (Cnidaria)

The phylum Cnidaria is an ancient group of venomous animals, specialized in the production and delivery of toxins. Many species belonging to the class Anthozoa have been studied and their venoms often contain a group of peptides, less than 10 kDa, that act upon ion channels. These peptides and their targets interact with high affinity producing neurotoxic and cardiotoxic effects, and even death, depending on the dose and the administration pathway. Zoanthiniaria is an order of the Subclass Hexacorallia, class Anthozoa, and unlike sea anemone (order Actiniaria), neither its diversity of toxins nor the in vivo effects of the venoms has been exhaustively explored. In this study we assessed some toxicological tests on mice with a low molecular weight fraction obtained by gel filtration in Sephadex G-50 from Zoanthus sociatus crude extract. The gel filtration chromatogram at 280 nm revealed two major peaks, the highest absorbance corresponding to the low molecular weight fraction. The toxicological effects seem to be mostly autonomic and cardiotoxic, causing death in a dose dependent manner with a LD₅₀ of 792 μg/kg. Moreover, at a dose of 600 μg/kg the active fraction accelerated the KCl-induced lethality in mice.     

Genomewide association for a dominant pigmentation gene in sheep

Most published genomewide association studies (GWAS) in sheep have investigated recessively inherited monogenic traits. The objective here was to assess the feasibility of performing GWAS for a dominant trait for which the genetic basis was already known. A total of 42 Manchega and Rasa Aragonesa sheep that segregate solid black or white coat pigmentation were genotyped using the SNP50 BeadChip. Previous analysis in Manchegas demonstrated a complete association between the pigmentation trait and alleles of the MC1R gene, setting an a priori expectation for GWAS. Multiple methods were used to identify and quantify the strength of population substructure between black and white animals, before allelic association testing was performed for 49 034 SNPs. Following correction for substructure, GWAS identified the most strongly associated SNP (s26449) was also the closest to the MC1R gene. The finding was strongly supported by the permutation tree‐based random forest (RF) analysis. Importantly, GWAS identified unlinked SNP with only slightly lower p‐values than for s26449. Random forest analysis indicated these were false positives, suggesting interpretation based on both approaches was beneficial. The results indicate that a combined analytical approach can be successful in studies where a modest number of animals are available and substantial population stratification exists.