Nitric oxide inducing function and intracellular movement of chicken interleukin-18 in cultured cells

To evaluate the characteristics of chicken interleukin-18 （ChIL-18） in different forms in vitro, the ChIL-18 full-length gene （ChIL-18-F） and the ChIL-18 presumed mature protein gene （ChIL-18-M） were cloned and inserted into the eukaryotic expression vector pCI, to construct recombinant pCI-ChIL-18-F and pCI-ChIL-18-M. The recombinant plasmids were then transferred into chicken splenic lymphocytes （CSLs）. Western blot showed that ChIL-18-F, with a molecular weight of 23.0 kDa, was produced in CSLs transfected by pCI-ChIL-18-F; ChIL-18-M, with a molecular weight of 19.5 kDa, was produced in CSLs transfected by pCI-ChIL-18-M. The nitric oxide （NO） level in the transfected CSLs and the culture medium at different time points was further examined under confocal microscopy using 4,5-diaminofluorescein staining. The results showed that both pCI-ChIL-18-F and pCI-ChIL-18-M groups showed significant increase in intracellular and extracellular NO production compared with pCI transfected control cells. These results suggest that both ChIL- 18-F and ChIL- 18-M could stimulate NO secretion in CSLs. To characterize the intracellular distribution of ChIL-18, ChIL-18-F and ChIL-18-M were each fused to the enhanced green fluorescent protein gene, and expressed in Vero cells. The results showed that the ChIL-18-F tended to the membranous region in Veto cells, while ChIL- 18-M did not. This indicates that the N-terminal 27 amino acid peptide helped ChIL-18 target to Vero cell membranes.     

Gene-gene interactions between interleukin-12A and interleukin-12B with the risk of brain tumor

Emerging evidence from preclinical and clinical studies has shown that interleukin-12 (IL-12) has some effectiveness against endogenously arising brain tumor. The aim of this study was to investigate interactions of IL-12A and IL-12B polymorphisms on the risk of brain tumor. We analyzed IL-12A rs2243115 and IL-12B rs3212227 polymorphisms in 170 patients with brain tumor and 222 healthy controls in a Chinese population using a polymerase chain reaction-restriction fragment length polymorphism assay and DNA sequencing method. Individuals carrying a G allele of IL-12A rs2243115 had a significantly higher risk of developing brain tumor compared with those carrying a T allele (odds ratio [OR]=2.01, 95% confidence interval [CI], 1.17-3.45). After stratification analysis according to tumor types, a similarly higher risk was detected in patients with glioma (OR=2.56, 95% CI, 1.25-5.21). When gene-gene interactions were examined, carriers at both loci rs2243115 TG/GG and rs3212227 AC/CC had a 2.62-fold increased risk of glioma compared with those with rs2243115 TT and rs3212227 AC/CC genotypes (OR=2.62, 95% CI, 1.05-6.50). This study provides evidence that IL-12A rs2243115 may be associated with the risk of brain tumor. Additionally, gene-gene interactions of IL-12A rs2243115 and IL-12B rs3212227 may contribute to brain tumor susceptibility.     

Long Non-Coding RNA H19 Promotes Glioma Cell Invasion by Deriving miR-675

H19 RNA has been characterized as an oncogenic long non-coding RNA (lncRNA) in breast and colon cancer. However, the role and function of lncRNA H19 in glioma development remain unclear. In this study, we identified that H19/miR-675 signaling was critical for glioma progression. By analyzing glioma gene expression data sets, we found increased H19 in high grade gliomas. H19 depletion via siRNA inhibited invasion in glioma cells. Further, we found H19 positively correlated with its derivate miR-675 expression and reduction of H19 inhibited miR-675 expression. Bioinformatics and luciferase reporter assays showed that miR-675 modulated Cadherin 13 expression by directly targeting the binding site within the 3′ UTR. Finally, introduction of miR-675 abrogated H19 knockdown-induced cell invasion inhibition in glioma cells. To our knowledge, it is first time to demonstrate that H19 regulates glioma development by deriving miR-675 and provide important clues for understanding the key roles of lncRNA-miRNA functional network in glioma.     

用日本血吸虫部分cDNA表达质粒文库免疫小鼠产生的保护性效果

寻找保护性效果更好的抗原分子一直是抗血吸虫病疫苗研发领域的热点和难点。国内外筛选日本血吸虫 (Ｓｃｈｉｓｔｏ ｓｏｍａｊａｐｏｎｉｃｕｍ)保护性抗原分子采取的主要策略有 2种 :一是通过构建血吸虫某一生活史ｃＤＮＡ文库 ,采用探针 (核酸或抗体 )从文库或文库表达产物中筛选出特异性抗原基因 ,再逐一评估其保护性效果。该策略的主要缺陷是筛选操作费时费力 ,且效率不高。另一途径则是根据已知的曼氏血吸虫或其它种 (株 )保护性抗原基因序列 ,通过ＰＣＲ或核酸探针等技术筛选与之同源的相应抗原分子 ,这一方法 (尤其是ＰＣＲ方法 )尽…     

IQ67 DOMAIN protein 21 is critical for indentation formation in pavement cell morphogenesis

In plants, cortical microtubules anchor to the plasma membrane in arrays and play important roles in cell shape. However, the molecular mechanism of microtubule binding proteins, which connect the plasma membrane and cortical microtubules in cell morphology remains largely unknown. Here, we report that a plasma membrane and microtubule duallocalized IQ67 domain protein, IQD21, is critical for cotyledon pavement cell(PC) morphogenesis in Arabidopsis. iqd21 mutation caused increased indentation wi...     

Building a better neonatal mouse model to understand infant respiratory syncytial virus disease

Background

Respiratory syncytial virus (RSV) is the number one cause of lower respiratory tract infection in infants; and severe RSV infection in infants is associated with asthma development. Today, there are still no vaccines or specific antiviral therapies against RSV. The mechanisms of RSV pathogenesis in infants remain elusive. This is partly due to the fact that the largely-used mouse model is semi-permissive for RSV. The present study sought to determine if a better neonatal mouse model of RSV infection could be obtained using a chimeric virus in which the F protein of A2 strain was replaced with the F protein from the line 19 clinical isolate (rA2-19F).

Methods

Five-day-old pups were infected with the standard laboratory strain A2 or rA2-19F and various immunological and pathophysiological parameters were measured at different time points post infection, including lung histology, bronchoalveolar lavage fluid (BALF) cellularity and cytokines, pulmonary T cell profile, and lung viral load. A cohort of infected neonates were allowed to mature to adulthood and reinfected. Pulmonary function, BALF cellularity and cytokines, and T cell profiles were measured at 6 days post reinfection.

Results

The rA2-19F strain in neonatal mice caused substantial lung pathology including interstitial inflammation and airway mucus production, while A2 caused minimal inflammation and mucus production. Pulmonary inflammation was characterized by enhanced Th2 and reduced Th1 and effector CD8⁺ T cells compared to A2. As with primary infection, reinfection with rA2-19F induced similar but exaggerated Th2 and reduced Th1 and effector CD8⁺ T cell responses. These immune responses were associated with increased airway hyperreactivity, mucus hyperproduction and eosinophilia that was greater than that observed with A2 reinfection. Pulmonary viral load during primary infection was higher with rA2-19F than A2.

Conclusions

Therefore, rA2-19F caused enhanced lung pathology and Th2 and reduced effector CD8⁺ T cell responses compared to A2 during initial infection in neonatal mice and these responses were exacerbated upon reinfection. The exact mechanism is unknown but appears to be associated with increased pulmonary viral load in rA2-19F vs. A2 infected neonatal lungs. The rA2-19F strain represents a better neonatal mouse model of RSV infection.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-015-0244-0) contains supplementary material, which is available to authorized users.     

Effects of the grafting on the fluorescence properties of CdTe nanocrystals

In this paper, we systematically investigated the influence of graft reagents having an amino or a carboxyl terminus with different chain lengths on the fluorescence properties of water‐soluble thioglycolic acid‐stabilized CdTe nanocrystals (TGA–CdTe). Strong enhancement effects of the grafting on the fluorescence intensity of TGA–CdTe were observed. The experiment results demonstrated that short‐chain‐length grafting can increase the fluorescence intensity of CdTe nanocrystals (NCs) better than long‐chain‐length grafting, and the grafting did not influence the emission wavelength of the CdTe NCs. The fluorescence intensity of the carboxyl‐grafted TGA–CdTe was more stable than that of the amino‐grafted TGA–CdTe at wide pH ranges (pH 5.1–10.0). Copyright © 2009 John Wiley & Sons, Ltd.