The Design and Construction of K11: A Novel α-Helical Antimicrobial Peptide

Amphipathic α-helical antimicrobial peptides comprise a class of broad-spectrum agents that are used against pathogens. We designed a series of antimicrobial peptides, CP-P (KWKSFIKKLTSKFLHLAKKF) and its derivatives, and determined their minimum inhibitory concentrations (MICs) against Pseudomonas aeruginosa, their minimum hemolytic concentrations (MHCs) for human erythrocytes, and the Therapeutic Index (MHC/MIC ratio). We selected the derivative peptide K11, which had the highest therapeutic index (320) among the tested peptides, to determine the MICs against Gram-positive and Gram-negative bacteria and 22 clinical isolates including Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Klebsiella pneumonia. K11 exhibited low MICs (less than 10 μg/mL) and broad-spectrum antimicrobial activity, especially against clinically isolated drug-resistant pathogens. Therefore, these results indicate that K11 is a promising candidate antimicrobial peptide for further studies

Effect of intravitreal injection of anti-VEGF on choroidal thickness and blood flow in posterior ciliary artery in patients with wet ARMD

AIM:To investigate the effect of intravitreal injection of anti-vascular endothelial growth factor(VEGF)on choroidal thickness and posterior ciliary artery blood flow in patients with wet age-related macular degeneration(ARMD). METHODS:A total of 93 wet ARMD patients enrolled into the ophthalmology department in our hospital from September 2014 to December 2016 were selected into our study. The patients were treated with anti-VEGF vitreous injection for 3mo. The central retinal thickness(CRT), subfoveal choroidal thickness(SFCT), best corrected visual acuity(BCVA, LogMAR), choroidal neovascularization(CNV)and parameters of ciliary arterial blood flow \〖systolic peak flow velocity(PSV), end-diastolic blood flow velocity(EDV)and mean blood velocity(mV), resistance index(RI)/pulse index(PI)\〗 before treatment(T0), 1mo after treated(T1), 2mo after treated(T2), 3mo after treated(T3)were recorded and compared. All patients had been followed for 18mo, and side-effect were recorded. RESULTS: All wet ARMD patients had successfully completed intravitreal anti-VEGF injection. Compared with before treatment, BCVA, CRT, SFCT, and CNV had gradually decreased(F=72.738, 32.333, 7.305 and 957.450, PPF=2.619, 1.942, 0.820, P>0.05). Compared with before treatment, RI index had gradually decreased(F=25.451, PF=8.660, PT2≈T1>T0; RI: T3≈T2CONCLUSION: Intravitreal injection of anti-VEGF can effectively reduce choroidal thickness and increase ciliary posterior arterial perfusion, with a good long-term effect and less risk

Immune responses of pigs inoculated with a recombinant fowlpox virus coexpressing ORF2/ORF1 of PCV2 and P1 2A of FMDV

A recombinant fowlpox virus (rFPV-ORF2ORF1-P12A) containing the open reading frame ORF2)/ORF1 DNAs of the porcine circovirus 2 (PCV2) (strain, Inner Mongolia) and foot-and-mouth disease virus (FMDV) capsid polypeptide of O/NY00 was evaluated for its abilities to induce humoral and cellular responses in piglets. In addition, we examined its abilities to protect cell cultures against a homologous virus challenge. To approach the feasibility of different united ways of immunization, the recombinant fowlpox virus rFPV-ORF2ORF1-P12A and the recombinant DNA plasmid pVAX1-IL18-ORF2ORF1 were used to immunize the pigs in “prime-boost”programme. We observed that priming the pigs with DNA plasmid pVAX1-IL18-ORF2-ORF1, followed by boosting with the recombinant virus rFPV-ORF2ORF1- P12A produced partially cellular immunity and humoral immunity. Control groups were inoculated with wild-type fowlpox virus (wtFPV) and phosphate buffer saline (PBS). All animals vaccinated with rFPVORF2ORF1- P12A developed specific anti-PCV2/anti-FMDV enzyme-linked immunosorbent assay (ELISA) and neutralizing antibodies and also showed T lymphocyte proliferation response. The antibody level produced by PCV2 was lower than that of O type FMDV to 1:20 and 1:200 respectively. We examined specific cytotoxic T lymphocyte (CTL) production in pigs serum and T lymphocytes (CD4, CD8, and CD4/CD8 double positive T cells) in the peripheral blood. First inoculating pVAX1-IL18- ORF2ORF1 and then rFPV-ORF2ORF1-P12A, had considerably higher CD4+, CD8+ and CD4+CD8+ T lymphocytes subgroups compared with the control groups. Whether the ratio between effective cells and target cells was 50:1 or 25:1, the specific CTL of experimental groups had much more significant differences with the control (FPV), even still the group of priming nucleic acid vaccine boosting recombinant virus had the bravest cytotoxicity of specific CTL. Moreover, the E/T ratio of 50:1 was more excellent. Following infection respectively with a mixture of a pathogenic strain of PCV2 (strain, Inner Mongolia)/FMDV (O/NY00) and neutralizing antibody, PK15 cells (BHK21) inoculated with recombinant fowlpox virus (rFPV) showed less (P < 0.05) yellow-green fluorescence and cytopathogenesis, suggesting the establishment of partial protection against PCV2/FMDV infection. The results show that the immunization programme here, in which pVAX1-IL18-ORF2ORF1 DNA vaccine was inoculated firstly and rFPV-ORF2ORF1-P12A was followed, is viable and indicates the potential use of a fowlpox virusbased recombinant vaccine for the control and prevention of PCV2/FMDV infections.Key words: PCV2, rFPV, FMDV, immune response, prime-boost

Lentinan relieves hepatitis B surface antigen induced functional impairment of monocytes/macrophages

Purpose: To investigate the efficacy of lentinan in relieving hepatitis B surface antigen (HBsAg)- induced functional impairment of monocytes/macrophages.Methods: Monocytic cell line THP-1 was incubated with lentinan and HBsAg for 24 h and then stimulated with LPS (Lipopolysaccharide). The expression levels of interleukin-1β (IL-1β), IL-12 and tumour necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent (ELISA) assays and quantitative reverse transcriptase-PCR (q-PCR). Protein levels of IkB-α, phospho-ERK, and phospho-p38 were measured by western blotting.Results: THP-1 cells treated with lentinan and HBsAg showed higher IL-1β, IL-12, and TNF-α levels than cells treated with HBsAg alone. The underlying mechanisms were associated with NF-kB and MAPK signal pathways. Decreased expression of IkB-α and phospho-ERK and increased expression of phospho-JNK and phospho-p38 were observed in cells treated with lentinan and HBsAg when compared with cells treated with HBsAg alone (p < 0.001). THP-1 cells incubated with 500 μg/mL lentinan secreted lower levels of cytokines than did control cells after LPS stimulation, suggesting an anti-inflammatory effect for lentinan.Conclusion: Lentinan shows both pro- and anti-inflammatory functions and may be a promising candidate for hepatitis B virus (HBV) treatment.Keywords: Hepatitis B surface antigen, Lentinan, Immuno-suppression, Pro-inflammatory, Antiinflammator

An In Vivo Screen Identifies PYGO2 as a Driver for Metastatic Prostate Cancer

Advanced prostate cancer displays conspicuous chromosomal instability and rampant copy number aberrations, yet the identity of functional drivers resident in many amplicons remain elusive. Here, we implemented a functional genomics approach to identify new oncogenes involved in prostate cancer progression. Through integrated analyses of focal amplicons in large prostate cancer genomic and transcriptomic datasets as well as genes upregulated in metastasis, 276 putative oncogenes were enlisted into an in vivo gain-of-function tumorigenesis screen. Among the top positive hits, we conducted an in-depth functional analysis on Pygopus family PHD finger 2 (PYGO2), located in the amplicon at 1q21.3. PYGO2 overexpression enhances primary tumor growth and local invasion to draining lymph nodes. Conversely, PYGO2 depletion inhibits prostate cancer cell invasion in vitro and progression of primary tumor and metastasis in vivo In clinical samples, PYGO2 upregulation associated with higher Gleason score and metastasis to lymph nodes and bone. Silencing PYGO2 expression in patient-derived xenograft models impairs tumor progression. Finally, PYGO2 is necessary to enhance the transcriptional activation in response to ligand-induced Wnt/β-catenin signaling. Together, our results indicate that PYGO2 functions as a driver oncogene in the 1q21.3 amplicon and may serve as a potential prognostic biomarker and therapeutic target for metastatic prostate cancer.Significance: Amplification/overexpression of PYGO2 may serve as a biomarker for prostate cancer progression and metastasis. Cancer Res; 78(14); 3823-33. ©2018 AACR

Opposing roles of TGFβ and BMP signaling in prostate cancer development

SMAD4 constrains progression of Pten-null prostate cancer and serves as a common downstream node of transforming growth factor β (TGFβ) and bone morphogenetic protein (BMP) pathways. Here, we dissected the roles of TGFβ receptor II (TGFBR2) and BMP receptor II (BMPR2) using a Pten-null prostate cancer model. These studies demonstrated that the molecular actions of TGFBR2 result in both SMAD4-dependent constraint of proliferation and SMAD4-independent activation of apoptosis. In contrast, BMPR2 deletion extended survival relative to Pten deletion alone, establishing its promoting role in BMP6-driven prostate cancer progression. These analyses reveal the complexity of TGFβ-BMP signaling and illuminate potential therapeutic targets for prostate cancer