Synthesis and preliminary antiproliferative evaluation of 1,3,9-triazacyclopenta[b]fluorene derivatives

Novel 1,3,9-triazacyclopenta[b]fluorene-4,10-diones and 1,3,9-triazacyclopenta[b]fluorene, analogue of ellipticine, were synthesised, and evaluated in vitro for their antiproliferative activity on various breast cancer cell lines.     

Potential anticancer activities of Rhus coriaria (sumac) extract against human cancer cell lines

Therapeutic strategies of plant origin are a better choice as both dietary plant products or its isolated active constituents against the development and progression of cancer. The present study aims to evaluate the anticancer activity of sumac (Rhus coriaria) against different human cancer MCF-7, PC-3, and SKOV3 cell lines. In addition, the study tries to explore a prospective mechanism of action, assessment of in vitro enzyme-inhibitory capacity of sumac extract against hCA I, II, IX, and XII. In the present study, the potential antitumor effects of sumac (Rhus coriaria) were explored in the human cancer cell lines; MCF-7, PC-3, and SKOV3 using in vitro assays. Apoptotic, cell survival, ELISA immunoassays were also conducted to reveal the inhibitory effects of sumac extract against hCA I, II, IX, and XII. In addition, both Clioquinol and Acetazolamide (AZM) were used as standards to explore the in vitro enzyme-inhibitory capacity of sumac extract against hCA I, II, IX, and XII. The hydro-alcoholic extract of R. coriaria (Sumac) was subjected to phytochemical analysis using GC/MS assays. Sumac at non-cytotoxic doses of 50 and 100 µM significantly modulates the growth of the MCF-7, PC-3, and SKOV3 cancer cells with a higher inhibitory effect and selectivity to carbonic anhydrase (CA) isoforms; hCA I, II, hCA IX, and XII. The data showed that sumac at doses of 50 and 100 µM significantly inhibited the growth, proliferation, and viability of cancer cells by activating the apoptotic process via caspase-3 overexpression and the regulation of Bcl-2 anti-apoptotic protein.     

新疆自然高温环境下玉米自交系开花期耐热性鉴定与评价

在新疆夏季自然高温环境下,以田间玉米空秆率、果穗结实率、相对结实率为主要评价指标,综合籽粒产量与穗部性状,对26份国内外玉米骨干自交系连续2年进行耐热性鉴定与评价,结合新疆当地高温干燥的气候特点,建立一套适宜玉米种质资源大田耐热性的鉴定、评价体系,为中国玉米耐热种质资源的遗传改良和新品种选育提供参考。结果表明:(1)不同基因型玉米自交系空秆率、果穗结穗率、相对结实率和耐热性差异较大,其中GW5F、GW4F、GW7F和PH6WC耐热性最强,在高温胁迫下空秆率最低,果穗结实率与相对结实率最高;其次为PHBA6、新自351等8份自交系,在高温胁迫下籽粒产量及综合表现较好;其余自交系对高温均表现敏感。(2)高温胁迫导致玉米穗部性状严重衰退,其中的穗重、穗行数和行粒数所受影响最大。(3)在年份或材料之间,玉米自交系籽粒产量与其主要农艺性状相关性差异较大,并以果穗重与籽粒产量相关性最高。(4)以相对结实率为主要指标通过层次聚类可将参试材料分为两大群,耐热性最强的GW5F、PH6WC、GW4M聚为第Ⅰ群,其余自交系聚为第Ⅱ群;第Ⅱ群中‘吉63’、Mo17等6份自交系可聚为第1亚群,PHBA6、LH82等6份自交系聚为第2亚群,郑58、新农育6390M等11份自交系聚为第3亚群。综合分析来看,该研究中表现极耐高温的4份自交系均为国外优异种质,可用于玉米耐高温基础研究和遗传改良;表现较强耐热性的8份品系多来自于新疆本地选育的材料,其遗传基础较为广泛,适应当地的高温环境,也是优异的耐热育种资源。     

玉米穗腐病抗性鉴定、遗传分析与分子机制*

玉米是我国主要农作物之一,其产量占全国谷物总产量三分之一左右,在国民生活中发挥了重要作用。玉米生长过程中受多种病害侵扰,其中穗腐病是一种由真菌导致的常见病害,目前已鉴定出40余种可诱发玉米穗腐病的病原菌。穗腐病不仅可造成玉米产量损失也导致籽粒品质严重下降,病原微生物所产生次生毒素更是危及人畜安全。目前穗腐病防治以化学方法为主,但是因此所造成的种植成本增加和环境污染问题日益突出,选育抗性品种成为防控玉米穗腐病最经济和安全有效的方法。玉米穗腐病抗性属于典型数量性状,国内外已有多个研究组通过建立玉米穗腐病抗性研究体系,开展玉米群体大规模穗腐病抗性鉴定工作,并筛选出一批抗病材料,这为玉米穗腐病抗性遗传改良奠定了材料基础。利用数量性状位点(quantitative trait locus, QTL)定位和全基因组关联分析(genome-wide association study, GWAS)等方法在玉米 1~10号染色体均检测到显著相关位点。尽管如此,玉米穗腐病抗性研究成果应用于生产的实例较少,生产上仍然缺乏综合性状优良且高抗穗腐病的玉米品种。这一方面是由于玉米穗腐病抗性遗传机制十分复杂、抗性基因克隆工作进展缓慢;另一方面为缺乏对玉米穗腐病抗性遗传研究进展进行系统总结,未有效开发分子标记用于分子育种所致。通过综述玉米穗腐病抗性遗传研究进展,汇总已定位QTL位点和显著关联SNP,构建一致性图谱和鉴定出定位热点区间,并进一步对比分析定位区间候选基因特征和转录组、代谢组研究进展,对促进玉米穗腐病抗性机制研究和玉米抗性育种均具有重要意义。     

5-氟尿嘧啶敏感与非敏感的人胃癌BGC-823细胞基因表达谱差异分析

利用化疗药物5-氟尿嘧啶(5-Fu)对胃癌细胞株进行筛选和诱导,建立具有耐药性的胃癌细胞株.与亲代细胞株对比生长特性及基因表达谱,初步探讨胃癌细胞的耐药机制.采用四唑盐比色法(MTT)测定5-Fu对胃癌细胞株BGC-823的半数抑制浓度(IC50);根据IC50设计5-Fu剂量,用大剂量的5-Fu逐渐递增间歇给药的方法,反复筛选,获得5株胃癌耐药细胞株.比较耐药细胞株和亲本细胞株的形态和生长特性,继而再用人类肿瘤基因芯片(human cancer arrays)比较测试亲本与耐药细胞株的基因表达谱差异;对其中189个上调基因和133个下调基因采用Gene Ontology中的BP(Biological Process)分析这些基因相关的功能,并用MAS2.0分析这些基因可能涉及的信号通路.结果提示,胃癌细胞株耐药(5-Fu)相关基因可能与转录因子信号转导、TNF受体介导的细胞凋亡和抗凋亡、细胞周期调节、细胞粘附及MAP激酶类的功能相关.对细胞周期、信号转导相关的6个基因采用定量PCR验证,结果与基因表达芯片结果一致.上述结果有利于进一步发现胃癌细胞的耐药基因和相关信号通路,探索抗胃癌治疗的耐药机制.     

动物活体成像生物发光稳定细胞株的构建

目的:构建组成型表达萤光素酶基因的载体,建立稳定高表达萤光素酶的MCF-7乳腺癌细胞株,检测其对细胞增殖的影响及在传代后的表达效果.方法:以载体pGIA.20为模板,PCR扩增萤火虫萤光素酶基因,将其克隆到载体pIRESpuro2上,将获得的pIRESpuro2-Luc经酶切和测序验证后,转染293T、MCF-7及ZR...     

Mapping QTLs for tissue culture response in soybean (Glycine max (L.) Merr.)

Hempseed is rich in polyunsaturated fatty acids (PUFAs), which have potential as therapeutic compounds for the treatment of neurodegenerative and cardiovascular disease. However, the effect of hempseed meal (HSM) intake on the animal models of these diseases has yet to be elucidated. In this study, we assessed the effects of the intake of HSM and PUFAs on oxidative stress, cytotoxicity and neurological phenotypes, and cholesterol uptake, using Drosophila models. HSM intake was shown to reduce H₂O₂ toxicity markedly, indicating that HSM exerts a profound antioxidant effect. Meanwhile, intake of HSM, as well as linoleic or linolenic acids (major PUFA components of HSM) was shown to ameliorate Aβ42-induced eye degeneration, thus suggesting that these compounds exert a protective effect against Aβ42 cytotoxicity. On the contrary, locomotion and longevity in the Parkinson’s disease model and eye degeneration in the Huntington’s disease model were unaffected by HSM feeding. Additionally, intake of HSM or linoleic acid was shown to reduce cholesterol uptake significantly. Moreover, linoleic acid intake has been shown to delay pupariation, and cholesterol feeding rescued the linoleic acid-induced larval growth delay, thereby indicating that linoleic acid acts antagonistically with cholesterol during larval growth. In conclusion, our results indicate that HSM and linoleic acid exert inhibitory effects on both Aβ42 cytotoxicity and cholesterol uptake, and are potential candidates for the treatment of Alzheimer’s disease and cardiovascular disease.     

Investigation of xenotropic murine leukemia virus-related virus (XMRV) in human and other cell lines

Xenotropic murine leukemia virus-related virus (XMRV) was discovered in human prostate tumors and later in some chronic fatigue syndrome (CFS) patients. However, subsequent studies have identified various sources of potential contamination with XMRV and other murine leukemia virus (MLV)-related sequences in test samples. Biological and nucleotide sequence analysis indicates that XMRV is distinct from known xenotropic MLVs and has a broad host range and cell tropism including human cells. Therefore, it is prudent to minimize the risk of human exposure to infection by evaluating XMRV contamination in cell lines handled in laboratory research and particularly those used in the manufacture of biological products. Nested DNA PCR assays were optimized for investigating XMRV gag and env sequences in various cell lines, which included MRC-5, Vero, HEK-293, MDCK, HeLa, and A549, that may be used in the development of some vaccines and other cell lines broadly used in research. The sensitivity of the DNA PCR assays was <10 copies in approximately 1.8 x 10⁵ cells equivalent of human DNA. The results indicated the absence of XMRV in the cell lines tested; although in some cases DNA fragments identified as cellular sequences were seen following the first round of PCR amplification with the env primer pair.     

Life or Death? A Physiogenomic Approach to Understand Individual Variation in Responses to Hemorrhagic Shock

Severe hemorrhage due to trauma is a major cause of death throughout the world. It has often been observed that some victims are able to withstand hemorrhage better than others. For decades investigators have attempted to identify physiological mechanisms that distinguish survivors from nonsurvivors for the purpose of providing more informed therapies. As an alternative approach to address this issue, we have initiated a research program to identify genes and genetic mechanisms that contribute to this phenotype of survival time after controlled hemorrhage. From physiogenomic studies using inbred rat strains, we have demonstrated that this phenotype is a heritable quantitative trait, and is therefore a complex trait regulated by multiple genes. Our work continues to identify quantitative trait loci as well as potential epigenetic mechanisms that might influence survival time after severe hemorrhage. Our ultimate goal is to improve survival to traumatic hemorrhage and attendant shock via regulation of genetic mechanisms and to provide knowledge that will lead to genetically-informed personalized treatments.