Chronic hepatitis B (CHB) is a major global health issue. The role of rare genetic variants in CHB has not been elucidated. We aimed to identify rare allelic variants predisposing to CHB. We performed exome sequencing in 50 CHB patients who had no identifiable risk factors to CHB and 40 controls who were healthy and hepatitis B surface antibody positive, but had never received hepatitis B vaccination. We selected six rare variant alleles and followed up their association with disease status by Sanger sequencing in a case-control study comprising 1728 CHB patients and 1636 healthy controls. The latter had either not been immunized with hepatitis B vaccine or had uncertain vaccination status. Our results showed that transmembrane protein 2 p.Ser1254Asn, interferon alpha 2 p.Ala120Thr, its regulator NLR family member X1 p.Arg707Cys and complement component 2 p.Glu318Asp were associated with CHB, with P values of <1.0×10(-7), 2.76×10(-5), 5.08×10(-5), 2.78×10(-4) and ORs of 2.45 , 4.08, 2.34 and 1.97 respectively. The combined P value was <2.0×10(-16). As there has been no indication of immunological functions for the associated gene, transmembrane protein 2, we further studied its expression by immunohistochemistry, real time PCR and western blotting. Our results showed that it was strongly expressed by healthy hepatocytes, but its expression was reduced in liver tissues with CHB, hepatitis B viral (HBV) genome-containing HepG2.2.15 cells, as compared with healthy liver tissues and non-HBV genome-containing HepG2 cells (P= 0.022 and 0.0036 respectively). CONCLUSION: We have identified four missense mutations associated with CHB, our results providing evidence for rare inborn genetic defects that contribute to increased host susceptibility to CHB.(HEPATOLOGY 2012.).

The perennial evergreen herb, Chimaphila japonica is found exclusively in East Asian temperate coniferous or sometimes in deciduous forests. By using the Fast Isolation by Amplified Fragment Length Polymorphism (AFLP) of Sequences Containing repeats (FIASCO) protocol, 20 microsatellite primer sets were identified in two wild populations. Of these primers, 16 displayed polymorphisms and 4 were monomorphic. The number of alleles per locus ranged from one to six among populations, values for expected and observed heterozygosities ranged from 0.000 to 0.848 and from 0.000 to 1.000, respectively. The new SSR markers will be useful in obtaining estimates of population-level genetic diversity and in phylogeographic studies of C. japonica.

Background:   To investigate changes of anterior segment morphology in primary angle-closure glaucoma after phacoemulsification using the Pentacam system. Design:   Prospective, interventional study, Zhongshan Hospital of Sun Yat-sen University. Participants:   Eighty-five eyes from 60 patients with primary angle-closure glaucoma undergoing phacoemulsification. Methods:  Intraocular pressure was measured by Goldmann applanation tonometry. Anterior segment morphology was assessed using the Pentacam camera. Main Outcome Measures:   The intraocular pressure, central anterior chamber depth, peripheral anterior chamber depth, anterior chamber volume, pupil diameter, and anterior chamber angle pre- and 3 months postoperative. Results:   A total 78 eyes of 55 patients with primary angle-closure glaucoma were included in the analysis. Thirty-two eyes (41.0%) had acute primary angle-closure glaucoma and 46 eyes (59.0%) had chronic primary angle-closure glaucoma. In both groups, statistically significant decreases in intraocular pressure and increases in anterior chamber volume, central anterior chamber depth, peripheral anterior chamber depth, and anterior chamber angle inferiorly, nasally, temporally, and, superiorly were observed at three months after phacoemulsification (all P < 0.001). Conclusions:  Phacoemulsification induces significant changes in anterior segment morphology in primary angle-closure glaucoma accompanied by a significant fall in intraocular pressure in the short term. © 2012 The Authors. Clinical and Experimental Ophthalmology © 2012 Royal Australian and New Zealand College of Ophthalmologists.

In HCT116 colorectal cancer cells, HeLa cervical cancer cells and HuH-7 hepatoma cells, miR-223 is expressed at a low level. Through infection with lentivirus containing miR-223 precursor, miR-233 was overexpressed in all these cells. Interestingly, the expression levels of FOXO1 mRNA and protein, and phosphorylation levels became significantly lower than those of their control. FOXO1 was down-regulated mainly in the cytoplasm, while the nuclear FOXO1 level became relatively high compared to the cytoplasm. As the unphosphorylated active form of FOXO1 increased in the cells, cyclin D1/p21/p27 were up-regulated at either mRNA or protein level. Proliferation of the cells was also greatly inhibited when miR-223 was over-expressed. Therein, our data suggest that miR-223 regulates FOXO1 expression and cell proliferation.

Soluble ligands are important targets for therapy of cancers and other diseases. Therapeutic monoclonal antibodies (mAbs) against such ligands block their interactions with corresponding receptors but do not enhance their removal from the circulation and can increase their half-lives because of the long half-lives of the antibodies. We have hypothesized that mAbs targeting two or more nonoverlapping epitopes on the same ligand could form oligomeric antibody-ligand complexes that can bind to cells expressing Fc gamma receptors (FcγRs) with high avidity leading to their fast and irreversible removal from the circulation. Insulin-like growth factor II (IGF-II) is an example of such ligands and an important target for human cancer therapy. We identified two mAbs, m610.27 and m630.3, which bound to nonoverlapping epitopes on IGF-II with nanomolar affinity, and generated a bispecific antibody, m660. m660 inhibited the interaction of human IGF-II (hIGF-II) with the human breast cancer cell line MCF-7, hIGF-II-mediated IGF receptor type I and insulin receptor phosphorylation, and cell growth. In the presence of hIGF-II, large complexes of m660 were formed that bound to FcγRII-expressing BJAB cells much more efficiently than the monospecific antibody-hIGF-II complexes and were presumably phagocytosed by phorbol 12-myristate 13-acetate-stimulated macrophage-like U937 cells. A mixture of m610.27 and m630.3 exhibited similar properties. To our knowledge, these mAbs are the first reported to target nonoverlapping epitopes on a cancer-related ligand and could represent a novel class of candidate therapeutics against cancers. This approach could also be used to irreversibly eliminate other disease-related soluble ligands.

A partial wave analysis of the pp[over ¯] mass-threshold enhancement in the reaction J/ψ→γpp[over ¯] is used to determine its J^{PC} quantum numbers to be 0^{-+}, its peak mass to be below threshold at M=1832_{-5}^{+19}(stat)_{-17}^{+18}(syst)±19(model)  MeV/c^{2}, and its total width to be Γ<76  MeV/c^{2} at the 90% C.L. The product of branching ratios is measured to be BR[J/ψ→γX(pp[over ¯])]BR[X(pp[over ¯])→pp[over ¯]]=[9.0_{-1.1}^{+0.4}(stat)_{-5.0}^{+1.5}(syst)±2.3(model)]×10^{-5}. A similar analysis performed on ψ(3686)→γpp[over ¯] decays shows, for the first time, the presence of a corresponding enhancement with a production rate relative to that for J/ψ decays of R=[5.08_{-0.45}^{+0.71}(stat)_{-3.58}^{+0.67}(syst)±0.12(model)]%.

A ternary polymer memory device based on a single polymer with on-chain Ir(III) complexes is fabricated by combining multiple memory mechanisms into one system. Excellent ternary memory performances-low reading, writing, and erasing voltages and good stability for all three states-are achieved.

It has been reported that OIC-A006, an osteogenically inducible compound, is able to promote osteogenesis in vitro and in vivo. In this study, we used a rabbit critical-sized segmental radial defect model (CSD)(15 mm) to analyse the osteogenic activity of OIC-A006 in non-cell-seeded tissue engineering bone substitutes. The scaffold carrier was bovine sintered bone "true bone ceramic (TBC)". OIC-A006 was delivered by PLGA (D,L-lactide-co-glycolide acid) microspheres. Drug-free PLGA microspheres and rhBMP-2-loaded PLGA microspheres were used as negative and positive control groups, respectively. Three kinds of composite were fabricated by coupling TBC, type-I collagen and the corresponding microspheres. The animals were randomly divided into 4 groups:(1) Group A: defect only,(2) group B:TBC/Collagen/drug-free-microspheres,(3) group C:TBC/Collagen/ OIC-A006-microspheres,(4) group D: TBC/Collagen/rhBMP-2-microspheres. The samples were analysed by histology, X-ray,microcomputed tomography (micro-CT), and biomechanical analyses. The results showed that OIC-A006 promoted bone regeneration to a remarkable extent. It is suggested that the application of OIC-A006 might be a valuable method in bone tissue engineering for healing large segmental defects of long bones. However, the biomechanical strength was a little inferior to that of BMPs.

Low environmental temperatures promote anthocyanin accumulation and fruit colouration by upregulating the expression of genes involved in anthocyanin biosynthesis and regulation in many fruit trees. However, the molecular mechanism by which fruit trees regulate this process in response to low temperature remains largely unknown. In this study, the cold-induced bHLH transcription factor gene MdbHLH3 was isolated from an apple tree and was found to interact physically and specifically through two regions (amino acids 1-23 and 186-228) at the N terminus with the MYB partner MdMYB1 (allelic to MdMYB10). Subsequently, MdbHLH3 bound to the promoters of the anthocyanin biosynthesis genes MdDFR and MdUFGT and the regulatory gene MdMYB1 to activate their expression. Furthermore, the MdbHLH3 protein was posttranslationally modified, possibly involving phosphorylation following exposure to low temperatures, which enhanced its promoter-binding capacity and transcription activity. Our results demonstrate the molecular mechanism by which MdbHLH3 regulates low temperature-induced anthocyanin accumulation and fruit colouration in apple. © 2012 Blackwell Publishing Ltd.

Although considerable effort has been devoted to the design of various nanoprobes for the fluorescent detection of multiple biomarkers in a single assay, they often suffer from emission-overlapping, owing to small Stokes shifts and wide emission spectra, which results in cross-talk and inaccurate quantification. Herein, we report the design and synthesis of a new nanoprobe for multienzyme detection with completely resolved emission peaks under single-wavelength excitation. The probe was assembled by attaching a cleavable peptide spacer, which was comprised from a matrix metalloproteinase-2 (MMP-2) substrate and a MMP-7 substrate, onto the surface of gold nanoparticles (AuNPs) through cysteine residues. A lanthanide complex, BCTOT-Eu(III)(BCTOT=1,10-bis(5'-chlorosulfo-thiophene-2'-yl)-4,4,5,5,6,6,7,7-octafluorodecane-1,3,8,10-tetraone), and 7-amino-4-methylcoumarin (AMC) were attached to the N terminus and the C terminus of the peptide, respectively. In the presence of one or both targeting enzymes, the substrate was cleaved and fluorescence resonance energy transfer (FRET) between the dyes and AuNPs was prohibited, thereby resulting in the dramatic fluorescence emission of dyes. Importantly, there was no emission cross-talk between the two dyes, thereby ensuring accurate detection of each enzyme. Based on this, the simultaneous fluorescence image of MMP-2 and MMP-7 was accomplished in living cells under single wavelength excitation. The apparent differences in the fluorescence imaging indicated distinct differences between the expression levels of MMPs between the human normal liver cells and the human hepatoma cells.