Hepcidin, an antimicrobial peptide, has a dual function including innate immunity and iron regulation. Here, based on the sequence of an EST database, we have isolated and characterized a hepcidin gene (referred to as CsHepcidin) from half-smooth tongue sole (Cynoglossus semilaevis). Analysis of the coding regions indicated CsHepcidin gene comprised 3 exons and 2 introns. The putative CsHepcidin showed a great similarity to other hepcidin orthologues, particularly with respect to its 24 aa signal peptide, typical RX(K/R)R motif and eight conserved cysteine residues in the mature cationic peptide. Phylogenic analysis indicated that CsHepcidin was a hepcidin 1-type peptide of acanthopterygians, with highly homologous with Solea senegalensis hepcidin. In C. semilaevis ontogeny, CsHepcidin mRNA was detected at a low level in unfertilized eggs, increased on 6 d after hatching, and decreased remarkably at metamorphic stage. CsHepcidin transcripts showed a constitutive basal expression in most of the tissues, especially in liver. Challenge with formalin-inactivated Vibrio anguillarum led to significantly up-regulations of CsHepcidin gene in liver, head kidney and spleen in time-dependent manners. Biological activity analysis showed that recombinant CsHEP exhibited direct antimicrobial activity against bacterial pathogens in vitro, particularly showed strong activity against the principal fish pathogens, V. anguillarum and Edwardsiella tarda. All these results suggest that CsHepcidin may be involved in the initial response to invasion of microbial pathogens. Further exploration to elucidate the role of CsHepcidin in iron regulation and embryogenesis in C. semilaevis are needed.

Checkpoint kinases, CHK1 and CHK2 are activated in response to DNA damage that results in cell cycle arrest allowing sufficient time for DNA repair. Agents which lead to abrogation of such checkpoints have potential to increase the efficacy of such as chemo- and radio-therapies. Thiophene carboxamide ureas (TCUs) were identified as inhibitors of CHK1 by high throughput screening. A structure-based approach is described using crystal structures of JNK1 and CHK1 in complex with 1 and 2, and of the CHK1-3b complex. The ribose binding pocket of CHK1 was targetted to generate inhibitors with excellent cellular potency and selectivity over CDK1and IKKβ key features lacking from the initial compounds. Optimization of 3b resulted in the identification of a regioisomeric 3-TCU lead, 12a. Optimization of 12a led to the discovery of the clinical candidate 4 (AZD7762), that strongly potentiates the efficacy of a variety of DNA-damaging agents in preclinical models.

High mobility group box 1 protein (HMGB1) was recently discovered to be a critical late-acting cytokine and innate immune-modulating factor in sepsis, but the potential role and mechanism of HMGB1 in adaptive immunity remains elusive. The present study demonstrated that HMGB1 had a dual influence on immune function of CD4(+) T lymphocytes. Low dose of HMGB1 had no effect on the proliferation activity of CD4(+) T lymphocytes, but the Th1 cytokines production was increased. In contrast, treatment with high amount of HMGB1 suppressed the proliferative response and induced Th2 polarization of CD4(+) T lymphocytes. We found that the expression of mitofusin-2 (Mfn2; also named hyperplasia suppressor gene), a member of the mitofusin family, was decreased in CD4(+) T lymphocytes when stimulated with high dose of HMGB1. Up-regulation of Mfn2 attenuated the suppressive effect of HMGB1 on CD4(+) T lymphocytes, which was associated with profound elevation of intracellular calcium concentration ([Ca(2+)](i)) and nuclear factor of activated T cells (NFAT) activity. These results indicate that HMGB1 have a direct role on adaptive immunity, and the decrease of Mfn2 expression may be a major cause of HMGB1-mediated immune dysfunction and Ca(2+)-NFAT signaling defect of CD4(+) T lymphocytes.

A series of new poly-functionalized fused naphthyridine derivatives were synthesized via a three-component reaction of aldehyde, 2-aminoprop-1-ene-1,1,3-tricarbonitrile and enaminone in EtOH using EtONa as a base. During these reaction processes, the domino construction of fused naphthyridine skeleton with concomitant formation of two new pyridine rings was readily achieved via base promoted three-component reactions in a one-pot operation. The procedures are facile, avoiding time-consuming and costly syntheses, tedious work-up and purifications of precursors.

BACKGROUND: and objective: The gene encoding angiotensin converting enzyme (ACE) is a promising candidate for lung cancer. We aimed to assess three well-characterized polymorphisms of the ACE gene (A-240T, I/D, A2350G) and lung cancer in Chinese people, and complete a meta-analysis of the association of I/D polymorphism with lung cancer. METHODS: and results: In our case-control study, a total of 684 patients with lung cancer and 602 age-matched controls were recruited. Genotyping was performed using polymerase chain reaction (PCR) and ligase detection reactions (LDR) techniques. Single-locus analysis indicated that carriers of the A-240T allele had a significantly increased risk for lung cancer under additive (odds ratio (OR)=1.2; 95% confidence interval (CI): 1.02-1.42; P=0.027) and recessive (OR=1.8; 95% CI: 1.24-2.63; P=0.002) models, and that DD genotype carriers were 1.97 times more likely to develop lung cancer (95% CI: 1.25-3.11; P=0.004) compared with those with the I allele under the recessive model. However, no significance was observed in further haplotype analysis (P>0.05). In a meta-analysis of ACE gene insertion-deletion (I/D) polymorphism from six studies with 1183 lung cancer patients and 1065 controls, we failed to detect any significant association (overall OR=1.09; 95% CI: 0.84-1.41). A low probability of publication bias was observed. CONCLUSIONS:: Our results suggest that ACE gene A-240T polymorphism might be a genetic marker for the development of lung cancer in Chinese people.

High mobility group box-1 protein (HMGB1) had been proved to induce maturation and activation of dendritic cell (DC), however, the endogenous changes and mechanisms underlying are unknown. Since endoplasmic reticulum stress (ERS) activates an adaptive unfolded protein response (UPR) that facilitates cellular survival and repair, we hypothesized that HMGB1 may regulate the function of DC by modulating ERS. In our study, HMGB1 stimulation induced significant ERS responses in DCs in a time- and dose-dependent manner, demonstrated by the up-regulation of a number of ERS markers. Gene silence of XBP-1 in splenic DCs decreased the levels of CD80, CD86 as well as major histocompatibility complex (MHC)-II expression and cytokine secretion after HMGB1 treatment, when compared with untransfected or nontargeting-transfected DCs (all P<0.05). Moreover, XBP-1 silenced DCs after treatment with HMGB1 failed to stimulate notable proliferation and differentiation of T cells, unlike normal DCs or nontargeting-transfected DCs (all P<0.05). Gene silence of XBP-1 resulted in down-regulation of the receptor for advanced glycation end products (RAGE) expression on the surface of splenic DCs induced by HMGB1 stimulation (P<0.05). These findings demonstrate an important role for ERS and its regulator XBP-1 in HMGB1-induced maturation and activation of DCs.

Growth hormone-releasing hormone receptor (GHRHR) plays a critical role in growth hormone (GH) synthesis, release and regulation in animals. The objective of this study was to investigate variations of the chicken GHRHR gene and their associations with growth and reproduction traits in 768 Beijing You chickens. Results revealed three single nucleotide polymorphisms (SNPs) in the promoter region of the gene (g.-1654A>G, g.-1411A>G and g.-142T>C). Association analysis revealed that the novel SNP g.-1654A>G had significant effects on chicken body weight at 7, 9, 11, 13, 17 weeks of age and the age of first egg as well as egg number at 32, 36 and 40 weeks. Significant association was also observed between g.-1411A>G and g.-142T>C with EN24. Moreover, the age of first egg was distinctly related with g.-142T>C (P < 0.05). Although significant statistical difference was not detected in GHRHR mRNA levels among genotypes of the SNPs (P > 0.05), strong expression variations of the gene were found between the ages 17 and 20 weeks in the population (P < 0.05). These results suggest that the three SNPs in the GHRHR promoter could be used as potential genetic markers to improve the growth and reproductive traits in chickens.

BACKGROUND AND PURPOSE:TCCFs are a common complication following craniofacial trauma and are usually treated by coils or detachable balloons. The use of the liquid embolic agent Onyx as the sole agent for the treatment of TCCFs has been rarely reported. Herein, we summarized the preliminary experience and effectiveness of treating TCCFs with Onyx in 23 patients.MATERIALS AND METHODS:From the 36 type A CCFs treated in our department between September 2005 and March 2011, a total of 23 posttraumatic direct CCFs were treated by using Onyx only via transarterial approach.RESULTS:Immediate postprocedural angiograms demonstrated complete occlusion in all patients. All the patients underwent a single procedure except 1 with bilateral TCCFs. Up to 24-month clinical and 3-month angiographic follow-ups revealed an ongoing complete occlusion without any complications.CONCLUSIONS:In this series, the use of Onyx for the transarterial embolization of TCCFs was feasible and effective. Associated adverse events were rare.

In spite of its recent achievements, the technique of single particle electron cryomicroscopy (cryoEM) has not been widely used to study proteins smaller than 100 kDa, although it is a highly desirable application of this technique. One fundamental limitation is that images of small proteins embedded in vitreous ice do not contain adequate features for accurate image alignment. We describe a general strategy to overcome this limitation by selecting a fragment antigen binding (Fab) to form a stable and rigid complex with a target protein, thus providing a defined feature for accurate image alignment. Using this approach, we determined a three-dimensional structure of an ∼65 kDa protein by single particle cryoEM. Because Fabs can be readily generated against a wide range of proteins by phage display, this approach is generally applicable to study many small proteins by single particle cryoEM.

Purpose: A new technique called "curvilinear approach" for prostate seed implantation has been proposed. The purpose of this study is to evaluate the dosimetric benefit of curvilinear distribution of seeds for low-dose-rate (LDR) prostate brachytherapy.Methods: Twenty LDR prostate brachytherapy cases planned intraoperatively with VariSeed planning system and I-125 seeds were randomly selected as reference rectilinear cases. All the cases were replanned by using curved-needle approach keeping the same individual source strength and the volume receiving 100% of prescribed dose 145 Gy (V(100)). Parameters such as number of needles, seeds, and the dose coverage of the prostate (D(90), V(150), V(200)), urethra (D(30), D(10)) and rectum (D(5), V(100)) were compared for the rectilinear and the curvilinear methods. Statistical significance was assessed using two-tailed student's t-test.Results: Reduction of the required number of needles and seeds in curvilinear method were 30.5%(p < 0.001) and 11.8%(p < 0.49), respectively. Dose to the urethra was reduced significantly; D(30) reduced by 10.1%(p < 0.01) and D(10) reduced by 9.9%(p < 0.02). Reduction in rectum dose D(5) was 18.5%(p < 0.03) and V(100) was also reduced from 0.93 cc in rectilinear to 0.21 cc in curvilinear (p < 0.001). Also the V(150) and V(200) coverage of prostate reduced by 18.8%(p < 0.01) and 33.9%(p < 0.001), respectively.Conclusions: Significant improvement in the relevant dosimetric parameters was observed in curvilinear needle approach. Prostate dose homogeneity (V(150), V(200)) improved while urethral dose was reduced, which might potentially result in better treatment outcome. Reduction in rectal dose could potentially reduce rectal toxicity and complications. Reduction in number of needles would minimize edema and thereby could improve postimplant urinary incontinence. This study indicates that the curvilinear implantation approach is dosimetrically superior to conventional rectilinear implantation technique.