Endocytic protein Pal1 regulates appressorium formation and is required for full virulence of Magnaporthe oryzae

Endocytosis plays key roles during infection of plant-pathogenic fungi, but its regulatory mechanisms are still largely unknown. Here, we identified a putative endocytosis-related gene, PAL1, which was highly expressed in appressorium of Magnaporthe oryzae, and was found to be important for appressorium formation and maturation. Deletion of PAL1 significantly reduced the virulence of M. oryzae due to defects in appressorial penetration and invasive growth in host cells. The Pal1 protein interacted and colocalized with the endocytosis protein Sla1, suggesting it is involved in endocytosis. The Δpal1 mutant was significantly reduced in appressorium formation, which was recovered by adding exogenous cAMP and 3-isobutyl-1-methylxanthine (IBMX). Moreover, the phosphorylation level of Pmk1 in Δpal1 was also reduced, suggesting Pal1 functions upstream of both the cAMP and Pmk1 signalling pathways. As a consequence, the utilization of glycogen and lipid, appressorial autophagy, actin ring formation, localization of septin proteins, as well as turgor accumulation were all affected in the Δpal1 mutant. Taken together, Pal1 regulates cAMP and the Pmk1 signalling pathway for appressorium formation and maturation to facilitate infection of M. oryzae.     

STARD3NL inhibits the osteogenic differentiation by inactivating the Wnt/β‐catenin pathway via binding to Annexin A2 in osteoporosis

Osteoporosis is one of the leading forms of systemic diseases related to bone metabolism in the world. STARD3 N‐terminal like (STARD3NL) showed robust association with osteoporosis‐related traits. Yet, the molecular functional mechanisms of STARD3NL in osteoblasts is still obscure. In this study, we demonstrated a high level of STARD3NL expression in the bone tissues from the patients with low bone mass and ovariectomized (OVX)‐induced osteoporotic mice. We identified Stard3nl as a potent factor that negatively and positively regulates osteoblast differentiation and cell proliferation, respectively. Furthermore, inhibition of Stard3nl induced β‐catenin gene expression and the nuclear translocation of β‐catenin, as well as Wnt signalling activities, contributing to the activation of Wnt/β‐catenin signalling. Mechanistic studies revealed that Stard3nl bound with Annexin A2 (Anxa2) to suppress β‐catenin expression, resulting into the suppression of Wnt signalling and downstream osteogenic differentiation. Moreover, adeno‐associated virus 9 (AAV9)‐mediated silencing of Stard3nl reversed bone loss in OVX‐induced osteoporotic mice by the injection into the knee joints. Collectively, our study revealed that Stard3nl suppressed osteogenesis via binding with Anxa2, resulting into the inactivation of Wnt signalling. It also highlights the preventive and therapeutic potential of STARD3NL as a specific and novel target for osteoporotic patients.     

Predicting arsenic bioavailability to hyperaccumulator Pteris vittata in arsenic-contaminated soils

Using chemical extraction to evaluate plant arsenic availability in contaminated soils is important to estimate the time frame for site cleanup during phytoremediation. It is also of great value to assess As mobility in soil and its risk in environmental contamination. In this study, four conventional chemical extraction methods (water, ammonium sulfate, ammonium phosphate, and Mehlich III) and a new root-exudate based method were used to evaluate As extractability and to correlate it with As accumulation in P. vittata growing in five As-contaminated soils under greenhouse condition. The relationship between different soil properties, and As extractability and plant As accumulation was also investigated. Arsenic extractability was 4.6%, 7.0%, 18%, 21%, and 46% for water, ammonium sulfate, organic acids, ammonium phosphate, and Mehlich III, respectively. Root exudate (organic acids) solution was suitable for assessing As bioavailability (81%) in the soils while Mehlich III (31%) overestimated the amount of As taken up by plants. Soil organic matter, P and Mg concentrations were positively correlated to plant As accumulation whereas Ca concentration was negatively correlated. Further investigation is needed on the effect of Ca and Mg on As uptake by P. vittata. Moreover, additional As contaminated soils with different properties should be tested.     

厄贝沙坦对人脐静脉内皮细胞株EA.hy926增殖、凋亡、VEGF mRNA表达的影响

目的:应用不同浓度厄贝沙坦对人脐静脉内皮细胞株EA.hy 926的增殖、凋亡生物学效应及血管发生主要基因VEGFmRNA的表达进行体外研究,探讨厄贝沙坦对内皮细胞的血管生成效应。方法:各种浓度厄贝沙坦对人脐静脉内皮细胞株EA.hy926共同孵育24 h。细胞增殖采用CCK8法分析,Annexin V/PI双染法检测细胞凋亡。RT-PCR验证VEGFmRNA的表达。结果:厄贝沙坦各浓度干预组细胞形态无明显变化,CCK8结果提示厄贝沙坦各干预组相比对照组细胞增殖活力增高(P<0.05),呈浓度非依赖性。流式细胞仪分析厄贝沙坦各浓度干预组细胞无明显凋亡。RT-PCR发现厄贝沙坦1×10-4,1×10-5,1×10-6mol/L浓度组VEGFmRNA表达增高(P<0.05)。结论:厄贝沙坦促进EA.hy926细胞株细胞增殖,上调VEGFmRNA的表达。这提示除了降压效应,血管紧张素受体拮抗剂在缺血性心脏病如慢性心力衰竭治疗中具有一定作用。     

The Influence of Selenium on Root Growth and Oxidative Stress Induced by Lead in Vicia faba L. minor Plants

The effect of selenium (Se) on Vicia faba L. minor roots subjected to lead (Pb) stress was studied by investigating root growth, root viability, and antioxidant enzyme activity. The experiments were carried out on plants grown for 2 weeks on Hoagland medium supplied with 50 μM Pb in the form of lead nitrate Pb(NO(3))(2) and/or Se concentrations of 1.5 and 6 μM in the form of sodium selenite Na(2)SeO(3). It was shown that Pb reduced the root growth and caused serious damage in the roots, which was accompanied by metal accumulation in these tissues. The exposition of roots to Pb led to significant changes in the biochemical parameters: the MDA and T-SH content and glutathione peroxidase (GSH-Px) activity increased but the guaiacol peroxidase (GPOX) activity decreased. Moreover, Pb intensified O(2)(·-) production in the roots. Selenium at a lower concentration alleviated Pb toxicity which was accompanied by a decreased O(2)(·-) production in the apical parts of roots and increased the T-SH content and GPOX activity. However, higher Se concentration intensified MDA and T-SH accumulation and GPOX and GSH-Px activity in Pb-treated plant roots. At low concentration, Se improved cell viability whereas at high concentration it was pro-oxidant and enhanced the lipid peroxidation and cell membrane injury.     

Genetic variation in IL28B is associated with the development of hepatitis B-related hepatocellular carcinoma

To evaluate the role of host IL28B (interleukin 28B; interferon lambda 3) single nucleotide polymorphisms (SNPs) in predicting hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) susceptibility, three SNPs in the IL28B gene (rs12979860C/T, rs8099917G/T and rs12980275G/A) were examined in 330 subjects (including 154 HBV-related HCC patients, 86 non-HCC patients with chronic hepatitis B (CHB), 43 HBV self-limited infections and 47 healthy controls). Notably, the frequency of CC homozygosity was 91.5% in healthy controls and 72.9% in CHB, the difference being statistically significant (χ(2) = 6.40, P = 0.01). The statistically difference was seen between healthy controls (91.5%) and HCC (74.7%) (χ(2) = 6.05, P = 0.01). However, this significant finding was not seen between HBV self-limited and healthy controls. Carriers of the minor T allele in rs12979860 had a higher risk of HCC compared with non-carriers (χ(2) = 4.44, P = 0.04). Haplotype analyses revealed significant association between haplotype C-T-A and healthy controls, but not with the HCC group (96.6 vs. 82.0%, χ(2) = 6.08, P = 0.01). Analyses of genotype combination and gene-gene interaction showed that there was a positive interaction between rs12979860 and rs12980275, with an OR rate of 11.79 (likelihood test, P = 0.04). Our results suggest that the IL28B rs12979860 C/T polymorphism might affect susceptibility to the chronic HBV infection and progression of HCC. Of note, the T allele and non-CC genotypes have strong predictive effect of increasing susceptibility of chronic HBV infection and HCC.     

An improved strategy based on RAPD markers efficiently identified 95 peach cultivars

DNA markers have useful applications in cultivar identification. A novel analysis approach called cultivar identification diagram (CID) was developed using DNA markers in the separation of plant individuals. This new strategy is less time- and cost-consuming, has reliable results, and was constructed for fingerprinting. Ten 11-mer primers were used to amplify the genotypes; all 95 peach genotypes (from the National Peach Germplasm Repository, in Nanjing, China) were distinguished by a combination of 54 primers. The utilization of the CID among these 95 peach cultivars was also verified by the identification of three randomly chosen groups of cultivars. This identification showed some advantages including the use of fewer primers and easy separation of all cultivars by the corresponding primers marked in the right position on the CID. This peach CID could provide the information to separate any peach cultivars of these 95, which may be of help to the peach industry in China and for the utilization of DNA markers to identify other plant species.     

Enzyme kinetic and molecular docking studies on the metabolic interactions of 1-hydroxy-2,3,5-trimethoxy-xanthone, isolated from Halenia elliptica D. Don, with model probe substrates of human cytochrome P450 enzymes

Halenia elliptica D. Don is a Tibetan herb and medicinal preparations containing Halenia elliptica have been commonly used for the treatment of hepatitis B virus infection in China. The metabolism of 1-hydroxy-2,3,5-trimethoxy-xanthone (HM-1) to its metabolites is mediated through cytochrome P450 enzymes. This study aimed to investigate the herb-drug interaction potential of HM-1 by studying its effects on the metabolism of model probe substrates of five major CYP450 isoforms in human liver microsomes. HM-1 showed moderate inhibitory effects on CYP1A2 (IC(50)=1.06μM) and CYP2C9 (IC(50)=3.89μM), minimal inhibition on CYP3A4 (IC(20)=11.94μM), but no inhibition on model CYP2D6 (dextromethorphan) and CYP2E1 (chlorzoxazone) probe substrates. Inhibition kinetic studies showed that the K(i) values of HM-1 on CYP1A2, CYP2C9 and CYP3A4 were 5.12μM, 2.00μM and 95.03μM, respectively. HM-1 competitively inhibited testosterone 6β-hydroxylation (CYP3A4) but displayed mixed type inhibitions for phenacetin O-deethylation (CYP1A2) and tolbutamide 4-hydroxylation (CYP2C9). Molecular docking study confirmed the inhibition modes of HM-1 on these human CYP isoforms.