Effects of Different Sources and Levels of Zinc on H2O2-Induced Apoptosis in IEC-6 Cells

Zinc has been shown to be an inhibitor of apoptosis for many years. The present study was designed to investigate effects of three zinc chemical forms on H₂O₂-induced cell apoptosis in IEC-6 cells via analysis of cell vitality, LDH activity, apoptosis percentage, caspase-3 activity, and Bcl-2, Bax, and caspase-3, -8, and -9 gene expression. Cells were divided into H₂O₂ and zinc sources+H₂O₂ groups, and there are three different zinc sources [zinc oxide nanoparticle (nano-ZnO), zinc oxide (ZnO), and zinc sulfate (ZnSO₄)] and three concentrations (normal = 25 μM, medium = 50 μM, and high = 100 μM) used in this article. In the present study, we found the striking cytotoxicity of H₂O₂ higher than 200 μM on cell vitality, LDH activity, and apoptosis percentage in the cells using five different concentrations (50, 100, 200, 400, and 800 μM) of H₂O₂ for 4 h. Moreover, we observed that cell vitality was increased, LDH activity and apoptotic percentage were decreased, and gene expression level of Bax and caspase-3 and -9 was markedly reduced, while gene expression level of Bcl-2 and ratio of Bcl-2/Bax were increased in normal concentration groups of nano-ZnO and ZnSO₄ compared with H₂O₂ group, but no significant difference was observed in caspase-8 gene expression. Furthermore, medium or, more intensely, high concentrations of nano-ZnO and ZnSO₄ enhanced H₂O₂-induced cell apoptosis. Compared with nano-ZnO and ZnSO₄, ZnO showed weakest protective effect on H₂O₂-induced apoptosis at normal concentration and was less toxic to cells at high level. Taken together, we proposed that preventive and protective effects of zinc on H₂O₂-induced cell apoptosis varied in IEC-6 cells with its chemical forms and concentrations, and maybe for the first time, we suggested that nano-ZnO have a protective effect on H₂O₂-induced cell apoptosis in IEC-6 cells.     

东北山地水生鞘翅目昆虫多样性的比较研究

调查了东北长白山自然保护区及辽宁省医巫闾山两地静水水体中水生甲虫．结果表明,长白山阔叶红松林生态系统水生甲虫比辽宁医巫闾山农林复合生态系统水生甲虫物种丰富的多．长白山水生甲虫共有１７个属２９个种,而辽宁医巫闾山水生甲虫只有１０个属１３个种．两地水生甲虫Ｓｈａｎｎｏｎ多样性指数分别为２．１２４、１．６４３,Ｓｈａｎｎｏｎ均匀度分别为１．２６０、０．６４１,两种不同生态系统中的水生甲虫物种多度分布较好地拟合于对数级数模型,均以少数物种为优势种,其中长白山自然保护区水生甲虫以Ｈｙｄｒｏｇｌｙｐｈｕｓｊａｐｏｎｉｃｕｓ、Ｈａｌｉｐｌｕｓｓｉｍｐｌｅｘ为优势种,辽宁医巫闾山水生甲虫则以Ｈｙｄｒｏｇｌｙｐｈｕｓｐｕｓｉｌｕｓ、Ａｇａｂｕｓｕｓｕｒｉｅｎｓｉｓ、Ａｇａｂｕｓｂｒａｎｄｔｉ占优势．     

Oxidative stress and NF-κB signaling are involved in LPS induced pulmonary dysplasia in chick embryos

Inflammation or dysbacteriosis-derived lipopolysaccharides (LPS) adversely influence the embryonic development of respiratory system. However, the precise pathological mechanisms still remain to be elucidated. In this study, we demonstrated that LPS exposure caused lung maldevelopment in chick embryos, including higher embryo mortality, increased thickness of alveolar gas exchange zone, and accumulation of PAS⁺ immature pulmonary cells, accompanied with reduced expression of alveolar epithelial cell markers and lamellar body count. Upon LPS exposure, pulmonary cell proliferation was significantly altered and cell apoptosis was inhibited as well, indicating a delayed progress of pulmonary development. LPS treatment also resulted in reduced CAV-1 expression and up-regulation of Collagen I, suggesting increased lung fibrosis, which was verified by Masson staining. Moreover, LPS induced enhanced Nrf2 expression in E18 lungs, and the increased reactive oxygen species (ROS) production was confirmed in MLE-12 cells in vitro. Antioxidant vitamin C restored the LPS induced down-regulation of ABCA3, SP-C and GATA-6 in MLE-12 cells. Furthermore, LPS induced activation of NF-κB signaling in MLE-12 cells, and the LPS-induced decrease in SP-C expression was partially abrogated by blocking NF-κB signaling with Bay-11–7082. Bay-11–7082 also inhibited LPS-induced increases of ROS and Nrf2 expression. Taken together, we have demonstrated that oxidative stress and NF-κB signaling are involved in LPS induced disruption of pulmonary cell development in chick embryos.     

Putative phosphorylation sites on WCA domain of HA2 is essential for <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> single nucleopolyhedrovirus replication

Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality.The Helicoverpa armigera single nucleopolyhedrovirus (HearNPv) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain,in which phosphorylation status are supposed to be critical in respect to actin polymerization.In the present study,two putative phosphorylation sites (232Thr and 250Ser) and a highly conserved Serine (245Ser) on the WCA domain of HA2 were mutated,and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome.Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at 245Ser can produce infectious virions,both 232Tbr and 250Ser mutations were lethal to the virus.However,actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus,which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway.     

White spot syndrome virus induces metabolic changes resembling the warburg effect in shrimp hemocytes in the early stage of infection

The Warburg effect is an abnormal glycolysis response that is associated with cancer cells. Here we present evidence that metabolic changes resembling the Warburg effect are induced by a nonmammalian virus. When shrimp were infected with white spot syndrome virus (WSSV), changes were induced in several metabolic pathways related to the mitochondria. At the viral genome replication stage (12 h postinfection [hpi]), glucose consumption and plasma lactate concentration were both increased in WSSV-infected shrimp, and the key enzyme of the pentose phosphate pathway, glucose-6-phosphate dehydrogenase (G6PDH), showed increased activity. We also found that at 12 hpi there was no alteration in the ADP/ATP ratio and that oxidative stress was lower than that in uninfected controls. All of these results are characteristic of the Warburg effect as it is present in mammals. There was also a significant decrease in triglyceride concentration starting at 12 hpi. At the late stage of the infection cycle (24 hpi), hemocytes of WSSV-infected shrimp showed several changes associated with cell death. These included the induction of mitochondrial membrane permeabilization (MMP), increased oxidative stress, decreased glucose consumption, and disrupted energy production. A previous study showed that WSSV infection led to upregulation of the voltage-dependent anion channel (VDAC), which is known to be involved in both the Warburg effect and MMP. Here we show that double-stranded RNA (dsRNA) silencing of the VDAC reduces WSSV-induced mortality and virion copy number. For these results, we hypothesize a model depicting the metabolic changes in host cells at the early and late stages of WSSV infection.     

蛇毒类凝血酶calobin在毕赤酵母中的表达

蛇毒类凝血酶是临床上防治血栓栓塞性疾病的有效药物。参照朝鲜蝮蛇(Agkistrodon caliginosus,Korean Viper)类凝血酶calobin基因序列(GenBank AccessionNo.U32937.1),将人工合成的calobin基因克隆到酵母表达载体pPICZαA,于毕赤酵母中表达,得到了分子量约为32kD的重组calobin蛋白,经甲醇诱导培养,表达产物可获得3.5g/L的高表达量。重组蛋白经过阴离子交换柱Q-Sepharose Fast Flow和分子筛Sephacryl-S-100凝胶过滤层析等纯化步骤进行了初步纯化。纯化后的重组calobin可以在纤维蛋白原平板上形成水解圈,经SDS-PAGE实验显示,重组蛋白能水解纤维蛋白原的Aα链,产生一条约40kD左右的降解带。在实验中未能发现重组calobin对纤维蛋白原的凝固作用。     

耐辐射奇球菌铁结合蛋白DRA0258的抗氧化功能

【目的】通过对极端环境耐受的耐辐射奇球菌Deinococcus radiodurans R1全基因组进行序列比对分析,获得具有铁储备蛋白Ferritin类似功能基序的未知功能蛋白DRA0258,采用分子生物学技术对该蛋白的功能和性质进行了验证和分析。【方法】首先对DRA0258进行克隆表达和纯化,并经络合物显色法测定蛋白上铁结合含量;通过三段连接敲除法构建dra0258突变株,检测突变株在双氧水协迫下的生存率、总抗氧化活性及过氧化氢酶活性;利用实时定量PCR检测突变株内抗氧化酶类及铁转运相关性调控蛋白的基因转录水平。【结果】经体内外蛋白铁含量检测证实DRA0258具有一定的铁结合能力;双氧水生存率实验表明dra0258的缺失导致细胞的抗氧化能力显著下降;过氧化氢酶活性、总抗氧化活性检测及抗氧化酶类的基因转录水平检测证实dra0258基因的缺失导致细胞内一些抗氧化基因转录水平下调,细胞的抗氧化应激系统受到损伤,并影响了一些铁调控网络蛋白的基因转录水平。【结论】本研究证实DRA0258是一种铁结合蛋白,该编码基因的缺失影响胞内铁转运系统并使细胞抗氧化能力下调。     

The computer program structure for assigning individuals to populations: easy to use but easier to misuse

The computer program Structure implements a Bayesian method, based on a population genetics model, to assign individuals to their source populations using genetic marker data. It is widely applied in the fields of ecology, evolutionary biology, human genetics and conservation biology for detecting hidden genetic structures, inferring the most likely number of populations (K), assigning individuals to source populations and estimating admixture and migration rates. Recently, several simulation studies repeatedly concluded that the program yields erroneous inferences when samples from different populations are highly unbalanced in size. Analysing both simulated and empirical data sets, this study confirms that Structure indeed yields poor individual assignments to source populations and gives frequently incorrect estimates of K when sampling is unbalanced. However, this poor performance is mainly caused by the adoption of the default ancestry prior, which assumes all source populations contribute equally to the pooled sample of individuals. When the alternative ancestry prior, which allows for unequal representations of the source populations by the sample, is adopted, accurate individual assignments could be obtained even if sampling is highly unbalanced. The alternative prior also improves the inference of K by two estimators, albeit the improvement is not as much as that in individual assignments to populations. For the difficult case of many populations and unbalanced sampling, a rarely used parameter combination of the alternative ancestry prior, an initial ALPHA value much smaller than the default and the uncorrelated allele frequency model is required for Structure to yield accurate inferences. I conclude that Structure is easy to use but is easier to misuse because of its complicated genetic model and many parameter (prior) options which may not be obvious to choose, and suggest using multiple plausible models (parameters) and K estimators in conducting comparative and exploratory Structure analysis.     

A global alfalfa diversity panel reveals genomic selection signatures in Chinese varieties and genomic associations with root development

Alfalfa (Medicago sativa L.) is an important forage crop worldwide. However, little is known about the effects of breeding status and different geographical populations on alfalfa improvement. Here, we sequenced 220 alfalfa core germplasms and determined that Chinese alfalfa cultivars form an independent group, as evidenced by comparisons of F_ST values between different subgroups, suggesting that geographical origin plays an important role in group differentiation. By tracing the influence of geographical regions on the genetic diversity of alfalfa varieties in China, we identified 350 common candidate genetic regions and 548 genes under selection. We also defined 165 loci associated with 24 important traits from genome-wide association studies. Of those, 17 genomic regions closely associated with a given phenotype were under selection, with the underlying haplotypes showing significant differences between subgroups of distinct geographical origins. Based on results from expression analysis and association mapping, we propose that 6-phosphogluconolactonase (MsPGL) and a gene encoding a protein with NHL domains (MsNHL) are critical candidate genes for root growth. In conclusion, our results provide valuable information for alfalfa improvement via molecular breeding.