基于网络药理学和分子对接探讨多基源品种石韦抗炎作用机制的异同

目的运用网络药理学和分子对接方法探讨多基源品种石韦抗炎作用机制的异同。方法首先对药物和疾病潜在靶点进行挖掘和筛选,选取药物靶点与疾病靶点的交集基因,根据网络拓扑参数筛选石韦、有柄石韦和庐山石韦有效成分抗炎的核心靶点。然后使用Cytoscape 3.6.1软件构建“成分-靶点-疾病”网络。通过DAVID数据库进行GO富集分析和KEGG通路分析。最后将筛选出的活性成分进行分子对接。结果筛选出石韦、有柄石韦和庐山石韦相同的潜在活性成分有蔗糖、绿莲皂苷元、β谷甾醇和香草醛酸;其相同的核心靶点有PTGS1、PRSS1、SREBF1、NOS3、AR等14个,相同的信号通路有胆碱能突触信号通路、神经活性配体-受体相互作用、钙信号通路。石韦发挥抗炎作用特有的潜在活性成分为山奈酚-3-阿拉伯呋喃糖苷、高丽槐素、槲皮素,4-o-葡萄糖苷、异热马酮、原儿茶酸、槲皮素,特有的核心靶点为PRKCA、ALOX5、RELA、AHSA1、CYP3A4等24个,以及特有的TNF信号通路、乙型肝炎、细胞凋亡、鞘脂信号通路、NF-κB信号通路等29个。有柄石韦发挥抗炎作用特有的潜在活性成分为植物醇、酞酸二乙酯、十六烷、二氢咖啡酸、癸醛、圣草酚、正十六烷酸、邻苯二甲酸二异丁酯,特有的核心靶点为LPL、NFE2L2、NQO1、GSK3B,以及特有的PPAR信号通路、催乳素信号通路、雌激素信号通路。庐山石韦发挥抗炎作用特有的信号通路为精氨酸生物合成、cGMP-PKG信号通路、丙氨酸、天冬氨酸和谷氨酸代谢。分子对接结果表明,活性成分均与关键靶点有较强的结合能力。结论石韦、有柄石韦、庐山石韦多基源品种抗炎的活性成分和作用机制相似性小、差异性较大。提示多基源品种石韦在临床抗炎有各自的特色,要区别对待。

道地中药材川贝母特异性PCR鉴别方法研究

为建立快速检测中药材川贝母成分的特异性PCR法,从NCBI数据库下载植物HMGR基因序列,根据同源性区域设计兼并引物,对川贝母及其易混品鳞茎基因组进行扩增。通过比对川贝母特异性DNA片段区域,设计引物扩增获得川贝母基原物种特异核酸片段。从川贝母基原鳞茎中提取DNA,设计特异引物对BMH-TF/BMH-TR,优化PCR扩增条件,通过特异性、灵敏度实验验证,建立特异性PCR检测体系。采用该方法从川贝母基原中扩增出一条120 bp左右条带,而非川贝母样品、空白对照在相同条件下无扩增条带,检测灵敏度为2 ng/μL,检测下限为4 ng。对10个市售药材的检测结果证明,该方法简便可行、重现性好。该PCR法可快速、准确鉴别川贝母基原,具有操作简便、快速、特异性高的优点。

Study on Genetic Diversity of Agronomic Traits and Genetic Relationships among Core Collections of Bitter Gourd

[Objective] This study aimed to investigate the genetic diversity of agronomic traits and genetic relationships among core collections of bitter gourd.[Method] Total 141 germplasms of bitter gourd were selected,and the genetic diversity of 13 agronomic traits was analyzed.In addition,total 46 core collections of bitter gourd were employed,and their genetic relationships were analyzed based on the phenotypic values and genotypic values of 5 agronomic traits,respectively.[Result] The genetic diversity analysis of agronomic traits showed that the genetic diversity indexes of the 4 qualitative traits of bitter gourd germplasms ranged from 0.46 to 1.34;the distribution of the 9 quantitative traits data was more dispersed with average coefficient of variation of 20.02%.The genetic relationship analysis showed that based on the phenotypic values and genotypic values of the 5 quantitative traits,the genetic distances among the 46 core collections of bitter gourd were different.Based on the genotypic values,the genetic distances among the 46 bitter gourd core collections ranged from 0.84 to 10.71.The 46 germplasms were divided into 17 groups with the rescaled distance of 8.5,which further classified the relationships among different germplasms.[Conclusion] This study will lay a solid foundation for the effective utilization of core collections and new variety breeding in bitter gourd.

Study on Origin and Evolution of Polyploids in Parakmeria Hu et Cheng Using LEAFY Gene

[Objective] This study aimed to explore the origin and evolution of poly- poids in Parakmeria Hu et Cheng through LEAFY gene clone and sequence analysis. [Method] In this study, LEAFY gene in Parakmeria species and its relative genera was cloned and sequenced using molecular biology methods. With reference to LEAFY gene sequence published by NCBI, the origin pattern of polypoids in Parakmeria was explored and reasons for the distribution layout of different polypoids were analyzed through sequence alignment and phylogenetic analysis. [Result] Different Magnoliaceae species can be distinguished using the LEAFY gene, and there was a length polymorphism found in the 3＋ end of the LEAFY gene, which can be used to divide Magnoliaceae plants of different species or in different genera, thus of high application value. [Conclusion] Most Parakmeria tetraploids are produced by polyploidization of homologous chromosomes, while Parakmeria hexaploids are chiefly produced by both polyploidization of homologous chromosomes and heterologous hybridization.

基于品质特征的贝母类药材品种分类研究

目的建立基于品质特征的贝母类药材品种分类。方法采用本草学、生态学、中药学、植物化学、中药鉴定学,以及结合生物计量学等方法开拓性地对中药贝母地理生态分布、性状及化学成分等品质特征进行研究,并对《中国药典》2015年版贝母药材进行分类。结果结果表明贝母类药材可分为"浙贝""川贝"2个系列,建议川贝母、平贝母、伊贝母归为川贝系,浙贝母、湖北贝母归为浙贝系。结论对于《中国药典》2015年版同类多基原品种中药的"分"与"合"及贝母资源的合理开发和利用提供依据。

No association between phosphatase and tensin homolog genetic polymorphisms and colon cancer

AIM: To investigate the association between single nucleotide polymorphisms (SNPs) in the phosphatase and tensin homolog (PTEN) tumor suppressor gene and risk of colon cancer. METHODS: We utilized a population-based casecontrol study of incident colon cancer individuals (n= 421) and controls (n = 483) aged ≥ 30 years to conduct a comprehensive tagSNP association analysis of the PTEN gene. RESULTS: None of the PTEN SNPs were statistically significantly associated with colon cancer when controlled for age, gender, and race, or when additionally adjusted for other known risk factors (P > 0.05). Haplotype analyses similarly showed no association between the PTEN gene and colon cancer. CONCLUSION: Our study does not support PTEN as a colon cancer susceptibility gene.

Characterization of the cDNA encoding a BPI/LBP homologue in venom gland of the hundred-pace snake Deinagkistrodon acutus

Bactericidal/permeability-increasing protein （BPI） and LPS-binding protein （LBP） play an important role in host defence. Current evidence shows that BPI/LBP may be widely existed in different cells and tissue types of animals. A full-length cDNA clone encoding a BPI/LBP homologue （dBPI）, 1757 bp in size, was characterized in venom gland of the hundred-pace snake Deinagkistrodon acutus. Its deduced amino acid sequence of 417 residues had 13.8% - 21.5% identity to BPI like 1 （BPIL1） and BPI like 3 （BPIL3） of other animals. Conserved cysteine residues which are involved in disulfide bond formation between the final strand of the N-terminal beta sheet and the long alpha helix of BPI are identified as Cys146-Cys183 of dBPI. Phylogenetic tree analysis showed that the BPI/LBP homologues formed five large clusters and dBPI was in a large cluster including BPIL1 and BPIL3. dBPI mRNA shows a tissue specific expression in venom gland. This is the first study to identify the cDNA encoding BPI/LBP homologues from reptiles [ Current Zoology 55 （5） ： 376 - 382, 2009].

Molecular Studies on Heat Shock Protein 70 Gene （Hsp70） Using ClustalW Multiple Sequence Alignment Analysis

Degenerate primers are particularly useful in amplifying homologous genes from different organisms. This paper describes a method for designing degenerate primers for a given multiple alignment of DNA sequences of Hsp70 gene family using ClustalW algorithm. The authors used an in silico approach to find a homology between more than one accession numbers of DNA sequences, X67711.2 was for Oryza sativa Hsp70, AY372071.1 was for Nicotiana tabacum Hsp70 and L41253.2 was for Lycopersicon esculentum Hsc70. The three accession numbers which were retrieved by the BLASTn program depend on their expected value （E-value）. Multiple sequence alignment was performed by ClustalW algorithm to produce a conserved blocks and determine the consensus region which had been used to produce the forward and reverse primer by the primer select module of DNAStar Lasergene V7 and In-Silco PCR module of FASTPCR program ver.4.0.8 was performed to detect the melting temperatures （Tm） and predict the PCR product size, The results of designed degenerate primer showed that there was a homology found between the designed primers and the DNA templates for the three accession numbers with at least 80% identity. The result of degenerate PCR showed that the three bands of the amplified PCR products of the three accession numbers were detected at the same molecular weight of marker （400 bp） with a difference about 15 pb compared to the in silco PCR product （385 pb）. In conclusion, this study focused on the importance of using the clustalW algorithm for designing the degenerate primer.

Detection of Herbicide-tolerant Canola by Multiplex PCR

[Objective]This study aimed to establish a multiplex PCR detection method of herbicide-tolerant canola.[Method] An endogenous reference gene(CruA) and three exogenous genes(T-CaMV 35 S, P-CaMV 35 S and pat) were selected for multiplex PCR. Specific primers were designed based on national standards or related literature. The annealing temperature, ratio of primer concentration and sensitivity of the established multiplex PCR system were optimized. The optimal multiplex PCR system was verified with known samples. [Result] The experimental results showed that the optimal annealing temperature of multiplex PCR was 58 ℃; the optimal ratio of primer concentration(μmol/L) was T-CaMV 35S: CruA: P-CaMV 35S: pat=0.1: 0.2: 0.2: 0.2;the detection sensitivity of the established multiplex PCR method was 0.3 ng. The amplified bands of known samples were completely consistent with the molecular characteristics. [Conclusion] This study provided a rapid, accurate and effective multiplex PCR technique for detection of herbicide-tolerant canola.

Polymorphisms of clip domain serine proteinase and serine proteinase homolog in the swimming crab Portunus trituberculatus andtheir associationwith Vibrio alginolyticus

Clip domain serine proteases （cSPs） and their homologs （SPHs） play an important role in various biological processes that are essential components of extracellular signaling cascades, especially in the innate immune responses of invertebrates. Here, polymorphisms of PtcSP and PtSPH from the swimming crab Portunus tritubereulatus were investigated to explore their association with resistance/ susceptibility to Vibrio alginolyticus. Polymorphic loci were identified using Clustal X, and characterized with SPSS 16.0 software, and then the significance of genotype and allele frequencies between resistant and susceptible stocks was determined by a Zz test. A total of 109 and 77 single nucleotide polymorphisms （SNPs） were identified in the genomic fragments of PtcSP and PtSPH, respectively. Notably, nearly half of PtSPH polymorphisms were found in the non-coding exon 1. Fourteen SNPs investigated were significantly associated with susceptibility/resistance to I1. alginolyticus （P〈0.05）. Among them, eight SNPs were observed in introns, and one synonymous, four non-synonymous SNPs and one ins-del were found in coding exons. In addition, five simple sequence repeats （SSRs） were detected in intron 3 of PtcSP. Although there was no statistically significant difference of allele frequencies, the SSRs showed different polymorphic alleles on the basis of the repeat number between resistant and susceptible stocks. After fiarther validation, polymorphisms investigated here might be applied to select potential molecular markers ofP. trituberculatus with resistance to I1. alginolyticus.

论中药分子鉴定的方法和原则

中药的准确鉴定涉及到人民生命安全和切身利益。传统的鉴定方法,即感官评价、显微鉴定和理化鉴定均存在不同程度的局限性,而分子鉴定则为中药的快速和准确鉴定带来了新的契机。为了使中药的分子鉴定得到更加广泛和有效的应用,应该高度重视相关标准、规范的制订。本文提出了中药分子鉴定的一些主要方法:1)真伪品鉴定:特异聚合酶链式反应法;2)正品和替代品鉴定:DNA条形码鉴定法;3)多基源鉴定:群体遗传学分析法;4)产地鉴别:分子谱系地理学分析法。经上述方法仍无法鉴别的贵重药材可进一步应用人类亲子鉴定的方法,开发特异微卫星标记进行进一步的鉴定。

Salinity tolerance in barley during germination——homologs and potential genes

Salinity affects more than 6%of the world’s total land area,causing massive losses in crop yield.Salinity inhibits plant growth and development through osmotic and ionic stresses;however,some plants exhibit adaptations through osmotic regulation,exclusion,and translocation of accumulated Na+or Cl-.Currently,there are no practical,economically viable methods for managing salinity,so the best practice is to grow crops with improved tolerance.Germination is the stage in a plant’s life cycle most adversely affected by salinity.Barley,the fourth most important cereal crop in the world,has outstanding salinity tolerance,relative to other cereal crops.Here,we review the genetics of salinity tolerance in barley during germination by summarizing reported quantitative trait loci(QTLs)and functional genes.The homologs of candidate genes for salinity tolerance in Arabidopsis,soybean,maize,wheat,and rice have been blasted and mapped on the barley reference genome.The genetic diversity of three reported functional gene families for salt tolerance during barley germination,namely dehydration-responsive element-binding(DREB)protein,somatic embryogenesis receptor-like kinase and aquaporin genes,is discussed.While all three gene families show great diversity in most plant species,the DREB gene family is more diverse in barley than in wheat and rice.Further to this review,a convenient method for screening for salinity tolerance at germination is needed,and the mechanisms of action of the genes involved in salt tolerance need to be identified,validated,and transferred to commercial cultivars for field production in saline soil.

The gap in research on polyploidization between plants and vertebrates:model systems and strategic challenges

Polyploidization via whole-genome duplications （WGD） is a common phenomenon in organisms. However, investigations into this phenomenon differ greatly between plants and animals. Recent research on polyploid plants illustrates the immediate changes that follow WGDs and the mechanisms behind in both genetic and epigenetic consequences. Unfortunately, equivalent questions remain to be explored in animals. Enlightened by botanical research, the study of polyploidization in vertebrates involves the identification of model animals and the establishment of strategies. Here we review and compare the research on plants and vertebrates while considering intrageneric or intraspecific variation in genome size. Suitable research methods on recently established poly- ploidy systems could provide important clues for under- standing what happens after WGDs in vertebrates. The approach yields insights into survival and the rarity of polyploidization in vertebrates. The species of Carassius and the allopolyploid system of goldfish × common carp hybridization appear to be suitable models for unraveling the evolution and adaptation of polyploid vertebrates.