Ze-Zhou Wang

Improvement of the immunity of pig to Hog cholera vaccine by recombinant plasmid with porcine interleukin-6 gene and CpG motifs

In order to observe the dosage-effect of recombinant pig interleukin-6 gene and CpG motifs on the immune responses of swine to vaccine, a novel recombinant eukaryotic VPIL6C plasmid was packed with chitosan nanoparticles (CNP) prepared by ionic cross linkage, which contains pig interleukin-6 gene and immunostimulatory sequence consisted of 11 CpG motifs. CNP-VRIL6C was then utilized to inoculate 30-day-old piglets intramuscularly at the dosage of 0.5, 1.0 and 1.5mg/per capita, respectively. Meanwhile, the piglets were injected with attenuated classical Hog cholera vaccine and designated as A1, A2 and A3 group. The blood was weekly collected from the piglets after vaccination to detect the changes of immunoglobulins, specific antibody, interleukins, IFN-γ and immune cells. The results were found that compared to those of the control piglets injected with VR1020-CNP, the content of IgG, IgA and IgM, specific antibodies, IL-2, IL-6 and IFN-γ significantly increased in the sera from the treated three groups from 14 to 70 days after vaccination (P<0.05); the number of T(H), T(C) and CD3(+) positive T cells raised obviously in the blood of VPIL6C treated piglets (P<0.05). Also the above immune indexes of A1 group were significantly lower to different extent in comparison with those of A2 and A3 group from 14 to 56 days post inoculation (P>0.05). Moreover, the lymphocytes also remarkably elevated in the treated groups (P<0.05). These indicate that VPIL6C entrapped with CNP is a novel effective adjuvant to boost the humoral and cellular immunity of pig to Hog cholera, implying its potentiality to enhance the resistance of pig against infectious diseases.

A Novel Broadband E-Plane Omni-Directional Planar Antenna

A novel broadband E-plane omni-directional planar antenna is designed and fabricated, which consists of four curved printed dipole radiators with eight parasitical strips. The impedance bandwidth is expanded by properly adding two parasitical strips along the boresight of each dipole radiator. A good omni-directionality is achieved at horizontal plane, and the measured variations of far field-patterns in the E-plane are less than ±1.5 dB, cross polarization levels of all directions in the E-plane better than −10 dB over the operating frequency range 5.1 ∼ 6.8 GHz with the reflection coefficients less than −10 dB. It is demonstrated experimentally that the proposed antenna is suitable for WLAN operation in the 5 GHz band, such as the 5.2 GHz (5.15 ∼ 5.35 GHz) and 5.8 GHz (5.725 ∼ 5.875 GHz) bands.

Promotion of immunity of mice to Pasteurella multocida and hog cholera vaccine by pig interleukin-6 gene and CpG motifs

A novel oligodeoxynuleotides containing 11 CpG motifs was synthesized and inserted into the VR1020 plasmid containing pig interleukin-6 (IL-6) gene (VPIL6) to construct recombinant plasmid, VPIL6C. The chitosan nanoparticles (CNP) were prepared by ionic cross linkage to entrap the VPIL6C (VPIL6C-CNP), VPIL6 (VPIL6-CNP) and CpG (CpG-CNP). 42-Day old female mice were divided into four groups and intramuscularly injected respectively with 6 pmol VPIL6C-CNP, VPIL6-CNP, CpG-CNP and VR1020-CNP along with the bivalent vaccines against the Pasteurellosis and hog cholera. The blood was weekly collected from mice after vaccination to detect the changes of immunoglobulins, specific antibodies, IL-2, IL-4, IL-6 and immune cells. 28 days after vaccination, the mice were orally challenged with virulent Pasteurella multocida. The results showed that in comparison with those of the control VR1020 group, the content of immunoglobulins, specific antibodies and interleukins significantly increased in the sera from the treated two groups (P<0.05). Meanwhile, the number of lymphocytes and monocytes also remarkably elevated in the treated groups (P<0.05). The immune responses of VPIL6C mice were notably stronger than those of VPIL6 and CpG group. The challenge results proved that the overall immunity was further promoted in the treated mice which resisted the challenge infection; while the control mice manifested evident symptoms and lesions, and died of infection. These suggested that VPIL6C-CNP could better promote the immunity and resistance of mice against Pasteurellosis than VPIL6-CNP and CpG-CNP, and facilitate the development of effective adjuvant to enhance the immunity of animal against infection.

Prognostic significance of CD24 and CD44 in breast cancer: a meta-analysis.

Objective Numerous studies have focused on the prognostic roles of CD24 and CD44 in breast cancer, but the results have been equivocal. The aim of this study was to gain better insight into the relationship between expression of CD24 and of CD44, either alone or in combination, and prognostic parameters in breast cancer. Methods Publications addressing the associations of CD24 or CD44 expression with survival outcome in breast cancer were selected for the meta-analysis according to defined criteria. Studies were pooled and odds ratios (ORs) or hazard ratios (HRs) were calculated. Publication bias and sensitivity analyses were also conducted. Results Sixteen studies comprising 5,697 breast cancer cases were included in our meta-analysis. Overall, CD24 overexpression was significantly associated with histological grade (OR = 1.52; 95% CI 1.12-2.06, p = 0.007), stage (OR = 1.74; 95% CI 1.27-2.40, p<0.001), shortened overall survival (HR = 1.48; 95% CI 1.21-1.80, p<0.001) and disease-free survival (HR 1.45, 95% CI 1.19-1.76, p<0.001), while no such association was observed when we limited our analysis to CD44 and CD44+/CD24- phenotypes. Subgroup analyses for CD24 according to the studies categorized by ethnicity, staining patterns and follow-up period were also conducted, and supported the stability of the prognostic role of CD24. Conclusions Our study demonstrated that the putative stem cell marker CD24 was significantly associated with worse survival based on the obtained data. In particular, CD24 may play a role in tumorigenesis and cancer progression. However, further large-scale studies are needed to confirm these findings.

Improvements in a 4-Element High Gain Directional UWB Antenna Array

In this paper, a novel serrated structure at the edges of the antipodal Vivaldi antenna is introduced. Compared with the Vivaldi antenna without serrated structure, the novel structure antenna has a better Front-to-Back ratio (F/B ratio). Additionally, when building a 4-element array with mirror image elements placed alternately along the H plane, the cross-polarization level at the boresight is reduced a lot compared to conventional array configuration. The design of the novel 4-element Vivaldi antenna array with mirror image elements described in this paper experimentally produces a gain with a range of 9∼15 dBi, F/B ratio more than 15 dB, and cross-polarization level at the boresight is reduced more than 10 dB from 3.5 GHz to more than 10.6 GHz with –10 dB return loss. The novel antenna array can be used for directional UWB systems.

Matching Network Design for a Monopole Antenna Using the Micro-Genetic Algorithm

The design procedure of a novel antenna matching network is proposed to overcome the narrowband characteristics of the monopole antenna. The matching network operating from 156 to 200 MHz is designed based on micro-genetic algorithm (micro-GA). Optional topology and component parameters of the matching network can be determined simultaneously by appropriately coding. The micro-GA iteratively guides a population of randomly selected design candidates toward the optimal solution. Measured results show that voltage standing wave ratio (VSWR) of the antenna system is less than 2.0 and transducer power gain is greater than 88.89% over the desired band.

Enhancement of immunity and resistance in mice by pig IL‐6 gene and CpG motifs encapsulated in chitosan nanoparticle

This study was conducted to explore the synergetic effect of a novel plasmid containing a porcine IL‐6 gene and CpG motifs on immunity of mice in order to develop an effective adjuvant to boost resistance against infection. The synthetic oligodeoxynucleotide containing 11 CpG motifs was inserted into the reconstructed VR1020 plasmid containing the pig IL‐6 gene (VRPIL6), designated VRIL6C, and then encapsulated in chitosan nanoparticles (CNP) prepared by ionic cross linkage, designated VRIL6C‐CNP. The 3‐week old mice were injected, respectively, with VRIL6C‐CNP, VRIL6‐CNP, CpG‐CNP and VR1020‐CNP to detect the changes of immunity. At 28 days post inoculation, the mice were challenged with virulent hemolytic serotype 2 Streptococcus to test their resistance against infection. The results showed that there was a significant increase in immunoglobulins and interleukins in mice receiving VRIL6C‐CNP compared with the control groups, as well as an increase in the lymphocytes and monocytes in the inoculated mice, so that the immunity was remarkably improved in the VRIL6C‐CNP group. The challenge provoked stronger immunity and protection against infection in the VRIL6C‐CNP group than in the control mice that manifested severe symptoms and lesions. This suggests that VRIL6C‐CNP could remarkably enhance the nonspecific immunity of mice, and facilitate the development of an effective immunopotentiator to promote the resistance of the animals against infection.

Knockdown expression of IL-10Rα gene inhibits PRRSV replication and elevates immune responses in PBMCs of Tibetan pig in vitro

Increase of interleukin-10 (IL-10) induced by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection has been intensely studied to inhibit the anti-viral responses of host pigs. Blockade of expression of IL-10 receptor (IL-10R) by RNA interference (RNAi) may relieve the immunosuppression caused by excessive IL-10 in PRRSV infection. The recombinant short hairpin expressing plasmid targeted to pig IL-10Rα was transfected into peripheral blood mononuclear cells of Tibetan pig (Tp-PBMCs) prior to PRRSV inoculation, then the replication of PRRSV and immune responses in Tp-PBMCs were evaluated. The recombinant interfering plasmid greatly decreased PRRSV yield. The transcriptional level of IL-10Rαwas obviously inhibited by recombinant interfering plasmid; the expression of IL-10 was also down-regulated, while that of TGF-β1 was not affected. Furthermore, the recombinant plasmid notably up-regulated the mRNA levels of TLR3, TLR7, IFN-α, IFN-γ, IL-2, IL-4, IL-12p40 and MyD88, while that of IL-8 was apparently decreased; In addition, cell viability of Tp-PBMCs was clearly enhanced by the interfering recombinant plasmid. Our results suggest that knockdown the expression of pig IL-10Rα can evidently inhibit the PRRSV infection and enhance the anti-viral immune responses of pig immune cells, which may be a promising way for preventing virus infection and developing new effective immune-regulator to strengthen the host immunity against PRRS.

Enhancement of immune response of piglets to PCV-2 vaccine by porcine IL-2 and fusion IL-4/6 gene entrapped in chitosan nanoparticles

In order to develop a novel effective immunomodulator to enhance pig resistance against post-weaning multi systemic wasting syndrome (PMWS), a recombinant plasmid co-expressing pig interleukin-2 (IL-2) and fusion interleukin-4/6 (IL-4/6) genes, designated VRIL4/6-2, was constructed and encapsulated in chitosan (CS) nanoparticles prepared by the ionotropic gelation method. Then 21-day old piglets were divided into two groups and intramuscularly injected respectively with VPIL4/6-2-CS and saline along with the porcine circovirus-2 (PCV-2) vaccine. The blood was collected from each piglet on days 0, 7, 14, 28, 56 and 84 after vaccination to assay the immunological changes. Content of IgG2a, CD4+, CD8+ T cells increased significantly in the sera or blood of piglets treated with VPIL4/6-2-CS (P<0.05). Furthermore, the expression level of IL-2, IL-4, IL-6, IL-15, TLR-2, TLR-7, Bcl-2, TNF-α, CD45 and STATs (STAT1, STAT2, STAT3, STAT4) genes were significantly elevated in the treated piglets respectively in different days after inoculation (P<0.05). The growth weight gain of the treated piglets was markedly improved in comparison with the controls (P<0.05). These indicate that VPIL-4/6-2 entrapped with chitosan nanoparticles is a safe and promising effective adjuvant to promote the immune response of pig to PCV-2 vaccination.

Improved expression of recombinant fusion defensin geneplasmids packed with chitosan-derived nanoparticlesand effect on antibacteria and mouse immunity

In order to develop a secure and competent technique to express the human immune gene for fighting infections, we cloned and expressed the BD2/3 using VR1020 (a eukaryotic expression plasmid). BD2/3 contains human β-defensin 2 (BD2) and human BD3. To explore safe and effective DNA delivery molecules in vitro and in vivo, the fusion genes of BD2/3 were used as an immune-labelled gene to verify transfection effectivness of modified chitosan (CS). Plasmid of VR1020-BD2/3 was packed with biomaterials: CS, average molecular weight: 25000D; polyethylene glycol-O-chitosan-polyethylenimine (PEG-O-CS-PEI); liposomes (LP); polyamine cationic liposomes (PCL); polyamine cationic liposomes of protamine (PCL-protamine) by ionotropic gelation. We observed that BD2/3 fusion gene showed high bioactivity in vitro and in vivo. The BD2/3 fusion protein inhibited the proliferation of bacteria (S. aureus, S. pneumoniae, P. aeruginosa and E. coli). The Kunming mice were immune to these nanoparticles and we analyzed their delivery efficiency and gene expression effect. BD2/3 results in multiple changes of innate and required immune system of mice. BD2/3 increases expression of IgG, IgG1, IgG2a, IL-2, IL-6, IFN-γ, as well as of lymphocytes and monocytes. Following challenge with virulent E. coli, CD4+ and CD8+ positive T-cell counts were highly elevated in the BD2/3 immunized mice, resulting in higher survival rates of mice. These results indicate that nanoparticles containing modified CS and BD2/3 are potentially safe and effective drugs in vivo to improve the immunity against bacterial infection and enhance innate immunity and adaptive immunity against infectious diseases.