Co-expression of apoptin (VP3) and antibacterial peptide cecropin B mutant (ABPS1) genes induce higher rate of apoptosis in HepG2 and A375 cell lines

The antibacterial peptide cecropin B mutant (ABPS1) gene has a broad range of antibacterial and  antiproliferative properties. Apoptin (VP3), a chicken anaemia virus-encoded protein is known to induce  apoptosis in human transformed cells. To explore drug combination in human tumor cells, apoptin and ABPS1 eukaryotic expression vector pIRES2-EGFP-apoptin and pIRES2-EGFP-ABPS1 were constructed and their expression effect individually and in combinations were studied in HepG2 and A375 cells. The vector pIRES2-EGFP-ABPS1 and pIRES2-EGFP-apoptin were transfected into tumor cells HepG2 and A375 by the  lipofectamine-mediated DNA transfection procedure. At 48 h post transfection, the apoptotic rate obtained by flow cytometry and the morphological changes under light and scanning electron microscope of tumor cells  were significant. In contrast, the microvilli on the surface of the control cells were disrupted, decreased and even disappeared. The cell membrane was injured and intracellular substances leaked out. Furthermore, our  results indicate that the apoptotic rates of apoptin (27.32% in HepG2 and 9.34% in A375 cells), were higher  than ABPS1 (23.79% in HepG2 and 8.33% in A375 cells). Moreover, the co-expression of Apoptin and ABPS1  showed higher apoptotic rates which were 27.66 and 10.33% in HepG2 and A375 cells respectively. However, the apoptotic rates obtained in HepG2 cells treated with apoptin and apoptin and ABPS1 together were closely  similar, but, not in A375 cells. Therefore, the results of the present study showed that the combination of  Apoptin and ABPS1 has synergistic effect in HepG2 and A375 cell lines.Keys words: Apoptin, ABPS1, apoptosis, co-expression, HepG2, A375

Simulation and experiment of ultrasonic-assisted grinding process for natural diamond

To improve the surface quality of natural diamond grinding, the ultrasonic vibration was introduced for composite grinding, and an inelastic collision theory model was established to calculate the amplitude of ultrasonic vibration. The effects of ultrasonic amplitude, grinding speed and grinding grain size on the material removal of natural diamond (100) crystal surface were studied, and the optimal process parameters were sought. The results show that the ultrasonic amplitude ranges from 3.1 to 8.7 μm in the direction of easy grinding of natural diamond (110) crystal surface and 2.9 to 9.1 μm in the direction of easy grinding of natural diamond (100) crystal surface. The optimal process combination parameters of natural diamond (100) crystal surface grinding obtained by orthogonal experiment are ultrasonic amplitude of 6.0 μm, grinding disk speed of 2 800 r/min, diamond grinding grain size code of M3/6. The surface roughness Ra of diamond (100) crystal surface after ultrasonic assisted grinding is 16.21 nm, which is 63.83% lower than that of traditional mechanical grinding. Ultrasonic assisted grinding of natural diamonds can achieve better surface quality than traditional mechanical grinding

Selective and Effective Binding of Pillar[5,6]arenes toward Secondary Ammonium Salts with a Weakly Coordinating Counteranion

The selective and effective binding of secondary ammoniums with a weakly coordinating tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (BArF) counteranion by per-ethylated pillar[5,6]arenes is reported. The construction of a first pillararene-based self-sorting system consisting of two wheels and two axles is also described

Unique epitopes recognized by monoclonal antibodies against HP-PRRSV: deep understanding of antigenic structure and virus-antibody interaction.

Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a member of the genus Arterivirus within the family Arteriviridae. N and GP3 proteins are the immunodominance regions of the PRRSV viral proteins. To identify the B-cell linear antigenic epitopes within HP-PRRSV N and GP3 proteins, two monoclonal antibodies (mAbs) against N and GP3 proteins were generated and characterized, designated as 3D7 and 1F10 respectively. The mAb 3D7 recognized only HuN4-F112 not the corresponding virulent strain (HuN4-F5). It also recognized two other commercial vaccines (JXA1-R and TJM-F92), but not two other HP-PRRSV strains (HNZJJ-F1 and HLJMZ-F2). The B-cell epitope recognized by the mAb 3D7 was localized to N protein amino acids 7-33. Western blot showed that the only difference amino acid between HuN4-F112-N and HuN4-F5-N did not change the mAb 3D7 recognization to N protein. The epitope targeted by the mAb 1F10 was mapped by truncated proteins. We found a new epitope (68-76aa) can be recognized by the mAb. However, the epitope could not be recognized by the positive sera, suggesting the epitope could not induce antibody in pigs. These results should extend our understanding of the antigenic structure of the N protein and antigen-antibody reactions of the GP3 protein in different species

Complete genomic characteristics and pathogenic analysis of the newly emerged classical swine fever virus in China

Abstract Background Classical swine fever (CSF) is one of the most devastating and highly contagious viral diseases in the world. Since late 2014, outbreaks of a new sub-genotype 2.1d CSF virus (CSFV) had caused substantial economic losses in numbers of C-strain vaccinated swine farms in China. The objective of the present study was to explore the genomic characteristics and pathogenicity of the newly emerged CSFV isolates in China during 2014–2015. Results All the new 8 CSFV isolates belonged to genetic sub-genotype 2.1d. Some genomic variations or deletions were found in the UTRs and E2 of these new isolates. In addition, the pathogenicity of HLJ1 was less than Shimen, suggesting the HLJ1 of sub-genotype 2.1d may be a moderated pathogenic isolate and the C-strain vaccine can supply complete protection. Conclusions The new CSFV isolates with unique genomic characteristics and moderate pathogenicity can be epidemic in many large-scale C-strain vaccinated swine farms. This study provides the information should be merited special attention on establishing prevention and control policies for CSF

Primer sets for the amplification of the ORF3 gene and its truncated fragments.

<p>In the F112-GP3 primers, the forward and reverse primers contain <i>Bam</i>HI and <i>Xho</i>I recognition sites, respectively (underlined). Other oligomeric nucleic acid fragments were annealed directively and be used for amino acids pursue absence, which also contain <i>Bam</i>HI and <i>Xho</i>I recognition sites (underlined). Bold font indicates termination codons.</p><p>Primer sets for the amplification of the ORF3 gene and its truncated fragments.</p

PRRSV strains cited in this study.

<p>PRRSV strains cited in this study.</p

Reactivity of mAbs 3D7 (B) and 1F10 (C) with HP-PRRSV HuN4 and vaccine strains in Marc-145 cells and transiently transfected 293T cells expressing expressing N and GP3 proteins.

<p>HuN4-F112 (attenuated in our laboratory), JXA1-R, and TJM-F92 are commercial vaccines used in China. HuN4, HNZJJ, and HLJMZ are HP-PRRSVs isolated by our laboratory. The two mAbs 3D7 and 1F10 recognized N and GP3 protein expressed by transiently transfected cells. 293T cells pCAGGS-transfected were used as a negative control. Marc-145 cells infected by PRRSV staining with the mAb 3F7 and 293T cells pCAGGS-N/GP3-transfected staining with the mAbs 2E9 and 4G5 were used as positive controls (A). Magnification 200×.</p