Biological impacts of 'hot-spot' mutations of hepatitis B virus X proteins are genotype B and C differentiated

AIM: To investigate the biological impacts of “hot-spot” mutations on genotype B and C HBV X proteins (HBx). METHODS: Five types of “hot-spot” mutations of genotype B or C HBV X genes, which sequentially lead to the amino acid substitutions of HBx as I127T, F132Y, K130M+V131I, I127T+K130M+V131I, or K130M+V131I+F132Y, respectively, were generated by means of site-directed mutagenesis. To evaluate the anti-proliferative effects, HBx or related mutants’ expression vectors were transfected separately to the Chang cells by lipofectamine, and the cells were cultured in hygromycin selective medium for 14 d, drug-resistant colonies were fixed with cold methanol, stained with Giemsa dyes and scored (increase of the colonies indicated the reduction of the anti-proliferation activity, and vice versa). Different types of HBx expression vectors were co-transfected separately with the reporter plasmid pCMVβ to Chang cells, which were lysed 48 h post-transfection and the intra-cellular β-galactosidase activities were monitored (increase of the β-galactosidase activities indicated the reduction of the transactivation activity, and vice versa). All data obtained were calculated by paired-samples t-test. RESULTS: As compared to standard genotype B HBx, mutants of I127T and I127T+K130M+V131I showed higher transactivation and anti-proliferative activities, while the mutants of F132Y, K130M+V131I, and K130M+V131I+F132Y showed lower activities. As compared to standard genotype C HBx, I127T mutant showed higher transactivation activity, while the other four types of mutants showed no differences. With regard to anti-proliferative activity, compared to standard genotype C HBx, F132Y and K130M+V131I mutants showed lower activities, and K130M+V131I +F132Y mutant, on the other hand, showed higher activity, while the mutants of I127T and I127T+K130M+V131I showed no differences. CONCLUSION: “Hot-spot” mutations affect the anti-proliferation and transactivation activities of genotype B and/or C HBx, and the biological impacts of most “hot-spot” mutations on HBx are genotype B and C differentiated.published_or_final_versio

A predictive continuum dynamic user-optimal model for a polycentric urban city

A predictive continuum dynamic user-optimal model is extended to investigate the traffic equilibrium problem for a polycentric urban city with multiple central business districts (CBDs). The road network within the city is assumed to be dense and can be viewed as a continuum in which travellers can choose their routes in a two-dimensional space. Travellers are assumed to choose their route to minimise the actual total cost to the destination (i.e. the CBD). The model consists of two parts: the conservation law part and the Hamilton–Jacobi part. The finite volume method is used to solve each part on unstructured meshes. Because the two parts are closely interconnected and have different initial times, solving the model can be treated as a fixed-point problem, which is solved using a self-adaptive method of successive averages. Numerical experiments for an urban city with two CBDs are presented to demonstrate the effectiveness of the model and the numerical algorithm.postprin

Cloning, expression and location of RNase9 in human epididymis

<p>Abstract</p> <p>Background</p> <p>Mammalian spermatozoa become fully motile and fertile during transit through the luminal fluid of the epididymis. At least 200 proteins are present in the epididymal lumen, but the potential roles of these luminal proteins in male fertility are unknown. Investigation of the function of these proteins will elucidate the mechanism of sperm maturation, and also provide new drug targets for male contraception. We cloned RNase9 from a human epididymis cDNA library for characterization and analysis of its functions.</p> <p>Findings</p> <p>It was predicted that human <it>RNase9 </it>gene was located on chromosome 14q11.2 and encoded a 205 amino acids protein with a signal peptide of 26 amino acids at the N-terminus. The protein had eight conserved cysteine residues characteristic of the RNase A family members and several potential post-translational modification sites.</p> <p>At the transcriptional level, <it>RNase9 </it>was expressed in a wide variety of tissues, and the expression was higher in men than in boys. <it>RNase9 </it>was localized to the post-equatorial region of the sperms' head. Immunofluorescence staining showed that RNase9 protein was present mostly in the epithelium of the epididymal tubule. Recombinant RNase9 had no ribonuclease activity. In addition, RNase9 had no detectable effect on sperm motility and fertilization as demonstrated by blocking spermatozoa with anti-RNase9 polyclonal serum.</p> <p>Conclusion</p> <p><it>RNase9 </it>is expressed in a wide variety of tissues. It is located on the post-equatorial region of the sperm head and the epithelium of epididymal tubule. Although <it>RNase9 </it>belongs to the RNase A family, it has no ribonuclease activity.</p

Genes Underlying Positive Influence Of Prenatal Environmental Enrichment And Negative Influence Of Prenatal Earthquake Simulation And Corrective Influence Of Chinese Herbalmedicine On Rat Offspring: Irf7 And Ninj2

Background: Prenatal environmental enrichment (EE) has been proven to positively affect but prenatal stress negatively influence the physiological and psychological processes in animals, whose trans-generational genetic mechanism remains unclearly defined. We aimed to investigate and find out key genes underlying the positive-negative effects derived from prenatal interventions.Materials and Methods: Pregnant rats were randomized into EE group (EEG), earthquake simulation group (ESG), herbal group (HG) received herbal supplements in feed after earthquake simulation, and control group (CG).Results: Light Box Defecation Test (LBDT) showed EEG offspring presented less fecal pellets than CG offspring, ESG’s more than CG’s, and HG’s less than ESG (p’s&lt;0.05). Open-field Test (OFT) score of EEG was higher than CG offspring, of ESG’s was lower than CG’s, and HG’s higher than ESG’s. Irf7 and Ninj were screened, which were up-regulated in EEG, down-regulated in ESG (FC&lt;0.5), and were neutralized in HG. Prenatal EE could positively promote the nervous system development, prenatal earthquake simulation could retard the nervous system development and Chinese herbal remedy (JKSQW) which could correct the retardation.Conclusion: The negative-positive prenatal effect could contribute to altered gene expression of Irf7 and Ninj2 which also could play a key role in the improving function of JKSQWfor the kidneys.Keywords: Prenatal stress; Earthquake simulation; Light Box Defecation Test; Open-field Test; Irf7; Ninj

Vertically aligned smooth ZnO nanorod films for planar device applications

The growth of smooth and continuous zinc oxide (ZnO) films, consisting of densely packed vertical ZnO nanorods with (002) crystal orientation on silicon substrates has been achieved in this work by a chemical solution method. These ZnO thin films have much stronger photoluminescence emission than those from discrete ZnO nanorods under identical conditions. Large area surface acoustic wave devices were fabricated on these films using conventional photolithography, and exhibited two well-defined resonant modes of the Sezawa wave and its harmonic mode

SHM benchmark for high-rise structures : a reduced-order finite element model and field measurement data

2012-2013 > Academic research: refereed > Publication in refereed journalVersion of RecordPublishe

Rapid quantification of semen hepatitis B virus DNA by real-time polymerase chain reaction

Aim: To examine the sensitivity and accuracy of real-time polymerase chain reaction (PCR) for the quantification of hepatitis B virus (HBV) DNA in semen. Methods: Hepatitis B viral DNA was isolated from HBV carriers' semen and sera using phenol extraction method and QIAamp DNA blood mini kit (Qiagen, Germany). HBV DNA was detected by conventional PCR and quantified by TaqMan technology-based real-time PCR (quantitative polymerase chain reaction (qPCR)). The detection threshold was 200 copies of HBV DNA for conventional PCR and 10 copies of HBV DNA for real time PCR per reaction. Results: Both methods of phenol extraction and QIAamp DNA blood mini kit were suitable for isolating HBV DNA from semen. The value of the detection thresholds was 500 copies of HBV DNA per mL in the semen. The viral loads were 7.5×10 7 and 1.67×10 7 copies of HBV DNA per mL in two HBV infected patients' sera, while 2.14×10 5 and 3.02×10 5 copies of HBV DNA per mL in the semen. Conclusion: Real-time PCR is a more sensitive and accurate method to detect and quantify HBV DNA in the semen. © 2005 The WJG Press and Elsevier Inc. All rights reserved.published_or_final_versio

Near-Infrared Super Resolution Imaging with Metallic Nanoshell Particle Chain Array

We propose a near-infrared super resolution imaging system without a lens or a mirror but with an array of metallic nanoshell particle chain. The imaging array can plasmonically transfer the near-field components of dipole sources in the incoherent and coherent manners and the super resolution images can be reconstructed in the output plane. By tunning the parameters of the metallic nanoshell particle, the plasmon resonance band of the isolate nanoshell particle red-shifts to the near-infrared region. The near-infrared super resolution images are obtained subsequently. We calculate the field intensity distribution at the different planes of imaging process using the finite element method and find that the array has super resolution imaging capability at near-infrared wavelengths. We also show that the image formation highly depends on the coherence of the dipole sources and the image-array distance.Comment: 15 pages, 6 figure

H-induced platelet and crack formation in hydrogenated epitaxial Si/Si <inf>0.98</inf>B <inf>0.02</inf>/Si structures

An approach to transfer a high-quality Si layer for the fabrication of silicon-on-insulator wafers has been proposed based on the investigation of platelet and crack formation in hydrogenated epitaxial Si Si0.98 B0.02 Si structures grown by molecular-beam epitaxy. H-related defect formation during hydrogenation was found to be very sensitive to the thickness of the buried Si0.98 B0.02 layer. For hydrogenated Si containing a 130 nm thick Si0.98 B0.02 layer, no platelets or cracking were observed in the B-doped region. Upon reducing the thickness of the buried Si0.98 B0.02 layer to 3 nm, localized continuous cracking was observed along the interface between the Si and the B-doped layers. In the latter case, the strains at the interface are believed to facilitate the (100)-oriented platelet formation and (100)-oriented crack propagation. © 2006 American Institute of Physics