中国鸭茅主栽品种DNA指纹图谱构建

利用SSR标记和SCoT标记构建了我国主栽的21个鸭茅品种的DNA指纹图谱。从180对SSR引物和80个SCoT引物中,筛选出多态性高、谱带清晰的SSR引物和SCoT引物各24个。24对SSR引物在供试材料中共检测到186个条带,其中多态性条带为175个,品种特异条带6个,平均多态性比率94.03%,多态性信息量均值0.845,Shannon指数变幅0.4479~0.6549,基因多样性指数变幅0.2946~0.4633,可鉴别的品种数2~21个;利用24个SCoT引物在供试材料中共检测到321个条带,其中多态性条带为249个,品种特异条带6个,平均多态性比率76.33%,多态性信息量均值0.907,Shannon指数变幅0.2588~0.6329,基因多样性指数变幅0.1695~0.4451,可鉴别的品种数1~21个;5对SSR引物和5个SCoT引物在10个品种上具有唯一特征谱带,最终综合各项指标筛选出5个引物(A01E14、A01K14、B03E14、D02K13和SCoT23)上的37个条带用于鸭茅品种DNA指纹图谱构建,数据库中每个品种均具有唯一DNA指纹编码,构建的DNA指纹数据可用于鸭茅品种真伪鉴定,为品种权保护提供了科学依据。     

ROS-Mediated Autophagy Induced by Dysregulation of Lipid Metabolism Plays a Protective Role in Colorectal Cancer Cells Treated with Gambogic Acid

Gambogic acid (GA), the main active component of gamboge resin, has potent antitumor activity both in vivo and in vitro. However, the underlying molecular mechanisms remain unclear. In this study, we found that GA could initiate autophagy in colorectal cancer cells, and inhibition of the autophagy process accelerated the effect of proliferative inhibition and apoptotic cell death induced by GA, implying a protective role of autophagy. Two-dimensional electrophoresis-based proteomics showed that GA treatment altered the expression of multiple proteins involved in redox signaling and lipid metabolism. Functional studies revealed that GA-induced dysregulation of lipid metabolism could activate 5-lipoxygenase (5-LOX), resulting in intracellular ROS accumulation, followed by inhibition of Akt-mTOR signaling and autophagy initiation. Finally, results using a xenograft model suggested ROS-induced autophagy protect against the antitumor effect of GA. Taken together, these data showed new biological activities of GA against colorectal cancer underlying the protective role of ROS-induced autophagy. This study will provide valuable insights for future studies regarding the anticancer mechanisms of GA.     

柠檬醛损伤黄曲霉线粒体生化机理的研究

应用生物化学方法并结合扫描电镜,研究柠檬醛掺入黄曲霉细胞,并通过损伤线粒体(Mt),导致抑制其生长的机理。结果表明,在药物致敏浓度时,菌丝体经该醛作用后,胞内Mt呈不规则增多,氧化还原反应系统受到破坏,与对照组相比,柠檬醛组的琥珀酸脱氢酶(Succinate Dehydrogenase,SDH)、苹果酸脱氢酶(Malate Dehydrogenase,MDH)活性分别呈不可逆下降271%和242%,随着药物浓度的升高,SDH、MDH的活性直至消失;以琥珀酸、α酮戊二酸和丙酮酸为底物时,线粒体呼吸速率分别下降24.1%、14.3%和36.1%,提示柠檬醛能使菌丝体DNA、RNA、脂类和蛋白质等生物合成受到抑制,促进细胞死亡。     

High throughput SNP genotyping with two mini-sequencing assays

Single nucleotide polymorphisms (SNPs) are veryimportant markers that can be used in many areas such asevolutionary genetics [1], disease-susceptibility genes[2,3], personalized medicine and forensics. Only about20% of human polymorphisms are length polymorphisms,whereas about 80% of human polymorphisms areSNPs. Kruglyak et al. [4] reported that there were about11,000,000 SNPs in the world population. There are many kinds of SNP genotyping technology[5,6]: some are only suitable to low …     

Cocirculation of two transmission lineages of echovirus 6 in jinan, china, as revealed by environmental surveillance and sequence analysis

Enterovirus environmental surveillance on sewage from the city of Jinan, Shandong Province, China, was initiated in 2008. Thirty echovirus 6 (E6) strains-1 in 2008 and 29 in 2010-were isolated and identified. Most E6 isolates (n = 21) came from the sewage collected on August 2010, revealing high local E6 activity at that time. Interestingly, the VP1 sequences of most isolates, even from the same sewage, were not identical. Phylogenetic analysis of VP1 sequences revealed two lineages for these isolates, with 78.0 to 80.0% nucleotide identities with one another, 94.8 to 100.0% identity within the major lineage, and 92.7 to 98.5% identity within the minor one. The VP1 sequences of environmental isolates, clinical isolates from 1998 to 2010, and global E6 were subjected to evolutionary analysis using Bayesian phylodynamic methods. The inferred E6 VP1 ancestral sequence dated back to 1901 (range, 1873 to 1928) and evolved with 7.047 × 10(-3) substitutions per site per year. Shandong E6 segregated into three clusters, and the two environmental lineages belonged to clusters A and C, which originated in 2003 and 1992, respectively. The antigenicity analysis via neutralization assay confirmed great antigenic differences between Shandong isolates and a prototype strain. These findings underscore the value of continuous environmental surveillance and genetic analysis to monitor circulating enteroviruses in the population and give further insight into E6 evolution.     

基于经血源性子宫内膜干细胞的中枢神经系统疾病治疗的研究进展

干细胞研究已成为当今生命科学领域中的前沿和热点问题,该研究为探讨胚胎发生、组织细胞分化以及基因表达调控等生物学问题提供了理想的模型,同时也为临床组织缺陷性疾病和遗传性疾病的细胞治疗和基因治疗开辟了新的手段。其中,经血源性子宫内膜干细胞(Menstrual blood-derived stem cells,MenSCs)来源丰富,具有多向分化潜能和较低的免疫排斥的特性,可以实现个体化治疗,是临床最具有应用优势的干细胞。脑与脊髓作为中枢神经系统,其损伤极为常见,致死率和致残率居各类创伤之首。与周围神经系统损伤相比,中枢神经受损后恢复较为困难,其治疗仍缺乏突破。而MenSCs的治疗有希望解决此难题,故结合近年来国内外对MenSCs的生物学特性及其对中枢神经系统疾病治疗的研究作一综述,从而为中枢神经系统疾病的治疗提供参考。     

Expression and functional characterization of the cancer-related serine protease, human tissue kallikrein 14

Human tissue kallikrein 14 (KLK14) is a novel extracellular serine protease. Clinical data link KLK14 expression to several diseases, primarily cancer; however, little is known of its (patho)-physiological role. To functionally characterize KLK14, we expressed and purified recombinant KLK14 in mature and proenzyme forms and determined its expression pattern, specificity, regulation, and in vitro substrates. By using our novel immunoassay, the normal and/or diseased skin, breast, prostate, and ovary contained the highest concentration of KLK14. Serum KLK14 levels were significantly elevated in prostate cancer patients compared with healthy males. KLK14 displayed trypsin-like specificity with high selectivity for P1-Arg over Lys. KLK14 activity could be regulated as follows: 1) by autolytic cleavage leading to enzymatic inactivation; 2) by the inhibitory serpins alpha1-antitrypsin, alpha2-antiplasmin, antithrombin III, and alpha1-antichymotrypsin with second order rate constants (k(+2)/Ki) of 49.8, 23.8, 1.48, and 0.224 microM(-1) min(-1), respectively, as well as plasminogen activator inhibitor-1; and 3) by citrate and zinc ions, which exerted stimulatory and inhibitory effects on KLK14 activity, respectively. We also expanded the in vitro target repertoire of KLK14 to include collagens I-IV, fibronectin, laminin, kininogen, fibrinogen, plasminogen, vitronectin, and insulin-like growth factor-binding proteins 2 and 3. Our results indicate that KLK14 may be implicated in several facets of tumor progression, including growth, invasion, and angiogenesis, as well as in arthritic disease via deterioration of cartilage. These findings may have clinical implications for the management of cancer and other disorders in which KLK14 activity is elevated.     

中药非药用部位栽培灵芝的活性成分及药效变化

【目的】以丹参(Salvia miltiorrhiza Bge.)、菊花(Chrysanthemum morifolium Ramat.)、桔梗(Ptatycodongrandiflorum A.DC.)3种中药材的非药用部位作为灵芝袋料栽培的原料,研究灵芝子实体活性成分及药效变化。【方法】试验比较测定了非药用部位配方组与常规组(常规组作为实验对照组,配方由玉米芯、棉籽壳等常规基质组成)的灵芝农艺性状,子实体多糖和三萜含量,并对各组灵芝进行了小鼠急性毒性试验及药效试验。【结果】结果表明,非药用部位栽培灵芝生物转化率接近或者高于常规组,生长周期有所延长;活性成分上,除了丹参组(SM.G)的三萜含量有所降低外,其余各组的活性成分较常规配方组(OF.G)均有提高,菊花组(CM.G)灵芝的多糖和三萜含量最高,分别为2.47%和0.79%。最大耐受量试验表明,非药用部位栽培的灵芝子实体的小鼠最大耐受量均为100 g/kg。溶血素试验和促睡眠试验中菊花组效果优于常规组灵芝;抗疲劳上,只有常规组灵芝表现出一定的抗疲劳功效,而非药用部位栽培的灵芝没有该药效。【结论】中药材非药用部位栽培灵芝是可行的,且子实体活性成分含量和部分药效发生了改变。     

Light-controlled inhibition of malignant glioma by opsin gene transfer

Glioblastomas are aggressive cancers with low survival rates and poor prognosis because of their highly proliferative and invasive capacity. In the current study, we describe a new optogenetic strategy that selectively inhibits glioma cells through light-controlled membrane depolarization and cell death. Transfer of the engineered opsin ChETA (engineered Channelrhodopsin-2 variant) gene into primary human glioma cells or cell lines, but not normal astrocytes, unexpectedly decreased cell proliferation and increased mitochondria-dependent apoptosis, upon light stimulation. These optogenetic effects were mediated by membrane depolarization-induced reductions in cyclin expression and mitochondrial transmembrane potential. Importantly, the ChETA gene transfer and light illumination in mice significantly inhibited subcutaneous and intracranial glioma growth and increased the survival of the animals bearing the glioma. These results uncover an unexpected effect of opsin ion channels on glioma cells and offer the opportunity for the first time to treat glioma using a light-controllable optogenetic approach.     

Perinatal exposure to bisphenol-A impairs learning-memory by concomitant down-regulation of N-methyl-d-aspartate receptors of hippocampus in male offspring mice

Bisphenol-A (BPA) has been shown to influence development of the brain and behaviors. The purpose of the present report was to investigate the effects of perinatal exposure to BPA on learning/memory and its mechanism of action, especially focusing on N-methyl-d-aspartate receptor (NMDAR). Perinatal maternal exposure to BPA at 0.5, 5, and 50 mg/kg/d significantly extended the escape length to find the hidden platform in Morris water maze, and BPA at 0.5 or 5 mg/kg/d markedly decreased the percentage of time spent in the quadrant where the platform had been during training both in postnatal day (PND) 21 and PND 56 mice. The results of passive avoidance test showed that the error frequency to step down from a platform after received footshock was significantly increased, and the latency of the step-down response onto the grid floor 24 h after received footshock was obviously reduced by exposure to BPA at 5 and 50 mg/kg/d (P < 0.01) in the PND 21 offspring or at 50 mg/kg/d in the PND 56 offspring (P < 0.01). Furthermore, perinatal exposure to BPA significantly inhibited the expressions of NMDAR subunits NR1, NR2A, and 2B in the hippocampus during the development stage, especially in PND 56 mice. The expressions of estrogen receptor beta (ERβ) in both PND 21 and PND 56 mice were markedly down-regulated by BPA at 0.5, 5, and 50 mg/kg/d. These results indicate that perinatal exposure to BPA affects normal behavioral development in both spatial memory and avoidance memory, and also permanently influences the behavior of offspring in adulthood. The inhibition of expressions of NMDAR subunits and ERβ in hippocampus during postnatal development stage may be involved.