Expression and signal regulation of the alternative oxidase genes under abiotic stresses

Plants in their natural environment frequently face various abiotic stresses, such as drought, high salinity, and chilling. Plant mitochondria contain an alternative oxidase （AOX）, which is encoded by a small family of nuclear genes. AOX genes have been shown to be highly responsive to abiotic stresses. Using transgenic plants with varying levels of AOX expression, it has been confirmed that AOX genes are im- portant for abiotic stress tolerance. Although the roles of AOX under abiotic stresses have been extensively studied and there are several excellent reviews on this topic, the differential expression patterns of the AOX gene family members and the signal regulation of AOX gene（s） under abiotic stresses have not been extensively summarized. Here, we review and discuss the current progress of these two important issues.     

Lenalidomide overcomes suppression of human natural killer cell anti-tumor functions by neuroblastoma microenvironment-associated IL-6 and TGFβ1

Background

Treatment for children with high-risk neuroblastoma with anti-disialoganglioside mAb ch14.18, IL-2, and GM-CSF plus 13-cis-retinoic acid after myeloablative chemotherapy improves survival, but 40 % of patients still relapse during or after this therapy. The microenvironment of high-risk neuroblastoma tumors includes macrophages, IL-6, and TGFβ1. We hypothesized that this microenvironment suppresses anti-tumor functions of natural killer (NK) cells and that lenalidomide, an immune-modulating drug, could overcome suppression.

Methods

Purified NK cells were cultured with IL-2, neuroblastoma/monocyte-conditioned culture medium (CM), IL-6, TGFβ1, and lenalidomide in various combinations and then characterized using cytotoxicity (direct and antibody-dependent cell-mediated cytotoxicity), cytokine, flow cytometry, and Western blotting assays. Anti-tumor activity of NK cells with lenalidomide, ch14.18, or both was evaluated with a xenograft model of neuroblastoma.

Results

CM from neuroblastoma/monocyte co-cultures contains IL-6 and TGFβ1 that suppress IL-2 activation of NK cell cytotoxicity and IFNγ secretion. IL-6 and TGFβ1 activate the STAT3 and SMAD2/3 pathways in NK cells and suppress IL-2 induction of cytotoxicity, granzymes A and B release, perforin expression, and IFNγ secretion. Lenalidomide blocks IL-6 and TGFβ1 activation of these signaling pathways and inhibits their suppression of NK cells. Neuroblastoma cells in NOD/SCID mice exhibit activated STAT3 and SMAD2/3 pathways. Their growth is most effectively inhibited by co-injected peripheral blood mononuclear cells (PBMC) containing NK cells when mice are treated with both ch14.18 and lenalidomide.

Conclusion

Immunotherapy with anti-tumor cell antibodies may be improved by lenalidomide, which enhances activation of NK cells and inhibits their suppression by IL-6 and TGFβ1.     

Establishment and characterization of fetal fibroblast cell lines for generating human lysozyme transgenic goats by somatic cell nuclear transfer

This study was performed to qualify goat fetal fibroblast (GFF) cell lines for genetic modification and somatic cell nuclear transfer (SCNT) to produce human lysozyme (hLYZ) transgenic goats. Nine GFF cell lines were established from different fetuses, and the proliferative lifespan and chromosomal stability were analyzed. The results suggested that cell lines with a longer lifespan had stable chromosomes compared with those of cells lines with a shorter lifespan. According to the proliferative lifespan, we divided GFF cell lines into two groups: cell lines with a long lifespan (GFF1/2/7/8/9; group L) and cell lines with a short lifespan (GFF3/4/5/6; group S). Next, a hLYZ expression vector was introduced into these cell lines by electroporation. The efficiencies of colony formation, expansion in culture, and the quality of transgenic clonal cell lines were significant higher in group L than those in group S. The mean fusion rate and blastocyst rate in group L were higher than those in group S (80.3 ± 1.7 vs. 65.1 ± 4.2 % and 19.5 ± 0.6 vs. 15.1 ± 1.1 %, respectively, P < 0.05). After transferring cloned embryos into the oviducts of recipient goats, three live kids were born. PCR and Southern blot analyses confirmed integration of the transgene in cloned goats. In conclusion, the lifespan of GFF cell lines has a major effect on the efficiency to produce transgenic cloned goats. Therefore, the proliferative lifespan of primary cells may be used as a criterion to characterize the quality of cell lines for genetic modification and SCNT.     

环境因子对东洞庭湖优势冬季水鸟分布的影响

东洞庭湖国家级自然保护区是我国乃至全球湿地水鸟的重要越冬栖息地,研究水鸟群落与环境因子之间的关系,对修复鸟类栖息地具有重要意义.2010和2011年冬季分别对东洞庭湖湿地冬季水鸟进行调查,采用典范对应分析(CCA)分析环境因子与优势水鸟分布的关系,采用偏CCA评估各环境因子的影响强度.结果表明： 到道路距离、到居民距离、斑块密度、植被类型数、水面面积和苔草面积对东洞庭湖冬季水鸟分布具有显著影响(P<0.05),其影响强度为苔草面积>斑块密度>到居民距离>植被类型数>水面面积>到道路距离,苔草面积和斑块密度对水鸟分布的影响达极显著水平(P<0.01),是影响东洞庭湖湿地冬季水鸟的主要环境因子;景观多样性和芦苇面积对水鸟分布的影响未达到显著水平(P>0.05).回归分析结果显示,环境因子对不同水鸟丰富度的影响不同,白琵鹭、罗纹鸭和黑腹滨鹬丰富度随苔草面积增大而减小,随斑块密度增大而增大;豆雁、白额雁、小白额雁和绿翅鸭丰富度随苔草面积增大而增大,随斑块密度增大而减小.东洞庭湖冬季水鸟分布是由以苔草面积、斑块密度为主的多环境因子综合作用的结果.     

SDS‐PAGE‐free protocol for comprehensive identification of cytochrome P450 enzymes and uridine diphosphoglucuronosyl transferases in human liver microsomes

Comprehensive identification of cytochrome P450 enzymes (CYPs) and uridine diphosphoglucuronosyl transferases (UGTs) in human liver microsomes (HLMs) was performed with an SDS‐PAGE‐free protocol. HLMs were solubilized with 5% v/v ionic liquid, 1‐butyl‐3‐methyl imidazolium tetrafluoroborate, followed by tryptic digestion, and 2D‐SCX‐RPLC‐ESI‐MS/MS (LTQ XL) analysis in triplicate. In total, 27 CYPs and 12 UGTs were confidently identified with average sequence coverage as 30.99 and 25.07%, average peptide number as 14 and 13, and average unique peptide number as 7 and 4, respectively. The highly similar isoforms of CYP3A, CYP2C, and CYP4F subfamilies could be unambiguously differentiated from each other, despite the fact that the sequence similarity of CYP2C9 and CYP2C19 is 91%. In addition, protein spectral count was used to approximately evaluate the relative abundance of identified CYPs and UGTs, and the results agreed with previous immunochemistry reports.     

短枝木麻黄种群苗期表型多样性评价

以来源于大洋洲原生种源区、亚洲原生种源区、亚洲引种次生区以及非洲引种次生区4个区域的20个种源短枝木麻黄种子和当年生幼苗为材料,通过种子千粒重以及幼苗苗高、地径、一级侧枝粗度、一级侧枝长度等7个性状对短枝木麻黄表型多样性进行了研究,以探讨种群苗期表型遗传差异,为短枝木麻黄早期遗传选择和遗传改良提供基本资料。结果表明:(1)短枝木麻黄种子千粒重在区域间和区域内种源间差异极显著,且千粒重具有显著的地理变异模式,随经度的增大而降低。(2)当年生幼苗苗高、地径在不同区域间及区域内种源间均存在极显著差异,其中泰国干东港种源幼苗生长最好(苗高76.6cm,地径4.64mm),而种源汤加的幼苗生长最差(苗高28.3cm,地径2.58mm)。(3)当年生幼苗一级侧枝粗度、一级侧枝长度、二级侧枝长度、每小枝节数和齿叶数在不同区域间及区域内种源间均存在极显著差异,其中齿叶数在区域间的变异系数最大(82.15%)。(4)通径分析表明,一级侧枝长度对苗高具有显著的正向影响作用,而一级侧枝粗度和二级侧枝长度对地径具有显著正向影响作用,它们可作为短枝木麻黄优良新品种筛选的参考因子。     

细枝木麻黄离体培养过程中愈伤组织和再生芽的细胞组织学观察

为探讨细枝木麻黄(Casuarina cunninghamianaMiq.)愈伤组织分化过程的细胞组织学,对离体培养条件下的愈伤组织进行扫描电子显微镜和石蜡切片观察,分析愈伤组织的细胞分裂、分化以及芽再生的发生过程。结果表明,新鲜外植体培养于愈伤组织诱导培养基上,伤口处的薄壁细胞开始脱分化,培养1周后形成明显的愈伤组织;继续培养2周后,胚性愈伤组织形成,且表层细胞启动分化形成芽原基;培养4周,可肉眼观察到胚性芽原基,数量增多并逐渐分化形成不定芽;培养至第6周,生成不定芽,并大量增殖和分化。因此,细枝木麻黄是通过愈伤组织分化形成胚状体的途径进行植株再生的,为建立细枝木麻黄组织培养高效再生体系提供了理论依据。     

Negative Affect Mediates Effects of Psychological Stress on Disordered Eating in Young Chinese Women

Background

The bi-relationships between psychological stress, negative affect and disordered eating has been well studied in western culture, while tri-relationship among them, i.e. how some of those factors influence these bi-relationships, has rarely been studied. However, there has been little related study in the different Chinese culture. This study was conducted to investigate the bi-relationships and tri-relationship between psychological stress, negative affect, and disordered eating attitudes and behaviors in young Chinese women.

Methodology

A total of 245 young Chinese policewomen employed to carry out health and safety checks at the 2010 Shanghai World Expo were recruited in this study. The Chinese version of the Perceived Stress Scale (PSS-10), Beck Depression Inventory Revised (BDI-II), Beck Anxiety Inventory (BAI), and Eating Attitude Test (EAT-26) were administered to all participants.

Principal Findings

The total scores of PSS-10, BDI-II and BAI were all highly correlated with that of EAT-26. The PSS-10 score significantly correlated with both BDI-II and BAI scores. There was no statistically significant direct effect from perceived stress to disordered eating (–0.012, 95%CI: –.038∼0.006, p = 0.357), however, the indirect effects from PSS-10 via affect factors were statistically significant, e.g. the estimated mediation effects from PSS to EAT-26 via depression and anxiety were 0.036 (95%CI: 0.022∼0.044, p<0.001) and 0.015 (95%CI: 0.005∼0.023, p<0.01), respectively.

Conclusions

Perceived stress and negative affects of depression and anxiety were demonstrated to be strongly associated with disordered eating. Negative affect mediated the relationship between perceived stress and disordered eating. The findings suggest that effective interventions and preventative programmes for disordered eating should pay more attention to depression and anxiety among the young Chinese female population.