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Due to the emergence of antibiotic resistance, bacteriophage therapy appears to be an ideal weapon to utilize against pathogenic bacteria. This study aimed to isolate, identify and characterize the lytic bacteriophage effective against the multidrug-resistant Acinetobacter baumannii clinical isolates. The isolated bacteriophage caused lysis by applying the double-layer agar technique on A. baumannii up to 99% in 18 hours of incubation at 37

Antimicrobial resistance (AMR) has arisen as a global concern in recent decades. Plant extracts used in combination with antibiotics are promising against AMR, synergistically. The purpose of this study was to evaluate the component of the bitter ginger (

Activity of plant essential oils and their fractions was evaluated against characterized isolates of antibiotic resistant Enterococcus faecalis recovered from diarrheic children. The isolates were confirmed by polymerase chain reaction (PCR) targeting 16S rRNA gene amplification followed by nucleotide sequencing and accession numbers retrieved were MW349990.1, MW349859.1, MW332122.1, MW356805.1, MW349975.1, MW349988.1, MW356790.1, MW356244.1, MW341593.1 and MW332549.1. These isolates were screened for antibiotic susceptibility to a wide range of antibiotic groups and mean zone of inhibition (ZOI) of all antibiotics were recorded. Antibacterial activity of plant essential oils (n=05) was checked against three antibiotic resistant isolates of E. faecalis. Three plant essential oils having higher ZOI including Cinnamomum verum, Syzygium aromaticum and Nigella sativa were used against resistant E. faecalis isolates to determine minimum inhibitory concentration (MIC). The lowest MIC observed was of S. aromaticum (11.39±3.94 mg mL

Objectives The objectives of this study were to evaluate P. Aeruginosa isolates from cancer patients for the phenotypic pattern of antibiotic resistance and to detect the gene responsible for virulence as well as antibiotic resistance. Methods A total of 227 P. aeruginosa isolates were studied and 11 antibiotics were applied for susceptibility testing. PCR detection of the genes BIC, TEM, IMP, SPM, AIM, KPC, NDM, GIM, VIM, OXA, toxA and oprI was done. Finally, the carbapenem resistant isolates were tested for phenotypic identification of carbapenemase enzyme by Modified Hodge test. Results The results showed that the isolates were resistant to imipenem (95%), cefipime (93%), meropenem (90%), polymixin B (71%), gentamicin (65%), ciprofloxacin (48%), ceftazidime (40%), levofloxacin (39%), amikacin (32%), tobramycin (28%) and tazobactum (24%). The PCR detection of the carbapenem resistant genes showed 51% isolates were positive for IMP, GIM and VIM, 38% for AIM and SPM, 30% for BIC, 20% for TEM and NDM, 17% for KPC and 15% for OXA. However, toxA and oprI genes were not detected. 154 carbapenem resistant isolates were found positive phenotypically for carbapenemase enzyme identification by Modified Hodge test. Conclusion The co-existence of multiple drug-resistant bodies and virulent genes has important implications for the treatment of patients. This study provides information about treating drug-resistant P. Aeruginosa and the relationship of virulent genes with phenotypic resistance patterns. .

Outbreaks of Pseudorabies (PR) by numerous highly virulent and antigenic variant Pseudorabies virus (PRV) strains have been causing severe economic losses to the pig industry in China since 2011. However, current commercial vaccines are often unable to induce thorough protective immunity. In this study, a TK/gI/gE deleted recombinant PRV expressing GM-CSF was developed by using the HDR-CRISPR/Cas9 system. Here, a four-sgRNA along with the Cas9

Aminoglycosides are used in empiric treatment of critically ill patients. Efficacy of aminoglycoside has been reduced due to dissemination of resistance. The aim of this study was to evaluate aminoglycoside resistance in cancer patients with pneumoniae. A total of 150 Bronchoalveolar lavage and Bronchial washing samples were collected from cancer patients. The samples were identified with standard microbiological procedures. Phenotypic susceptibility pattern of the isolates was determined against various groups of antibiotics such as Penicillins, Cephalosporins, Carbapenems, Monobactams, Aminoglycosides, Tetracyclins, Glycopeptides and Sulphonamides. The isolates with phenotypic resistant to aminoglycosides were further evaluated for the presence of armA gene. The strains of E. coli (12.5%), S. aureus (15.6%), Streptococcus (15.6%), Pseudomonas (18.7%) and K. pneumoniae (37.5%) were isolated. The phenotypic resistance profile showed highest resistance against aminoglycosides (Tobramycin, 53.1% Gentamicin and 50% Amikacin) followed by cephalosporins and sulfonamides group. The armA gene was detected in aminoglycoside resistant isolates. The overall genotypic resistance was evaluated as 21.8%. The armA gene was found in K.pneumoniae 23.5%, Pseudomonas 11.8% (4/24) and E. coli 5.9%. High level resistance to aminoglycosides raises therapeutic concern to health care professionals. These findings highlight the importance of effective monitoring and surveillance to the use of broad-spectrum antibiotics.

Background: Dengue virus (DENV) is an RNA virus belonging to the family Flaviviridae of the genus Flavivirus with worldwide distribution. Dengue fever is caused by any of four closely-related serotypes DENV, an emerging pandemic-prone viral disease in many regions of the world. Objectives: The present study aimed to determine the prevalence of dengue virus genotypes and serotypes in children aged below 15 years in Lahore, Pakistan. Methods: In this study, 112 serum samples were collected from clinically suspected dengue fever patients from March 2017 to December 2018 at different tertiary care hospitals in Lahore, Pakistan. Regarding the patients’ age, the samples were divided into four groups from A to D (i.e., 0 - 1, 1 - 5, 5 - 10, and 10 - 15 years of age). Rapid immuno-chromatography (ICT) test was conducted on the collected serum samples, followed by quantitative RT-PCR for serotype of dengue virus. Results: Out of 112 samples, 34 samples were diagnosed as DENV positive by the rapid ICT screening method. No virus was detected in groups A and B, while three samples were positive in group C (1 boy and two girls), and 31 samples (23 boys and 8 girls) were positive in group D. The results of quantitative RT-PCR exclusively showed DEN-3 serotype in all the ICT positive samples. The results indicated that the prevalence of DEN-3 serotype in children was 100%, indicating that DEN-3 serotype might cause severe epidemics in the future in Lahore, Pakistan. Hematological analysis revealed an increase in hematocrits in 41.1% dengue-positive cases. Leucopenia was prominent in 79.4% of the cases, while Thrombocytopenia was reported in 70.5% of the participants. The biochemical analysis also indicated an increase in liver enzymes in patients (ALT 88%, AST 79%), while the lower levels of cholesterol (69 %) and serum albumin (25%) were also observed. Conclusions: Dengue virus spreads and grows quickly worldwide over a highly short time interval. Dengue fever claims for a significant number of lives. This study would help individuals know about the status of laboratory parameters in dengue fever and detect how to overcome the prevalence of Dengue virus.

Background: Silver nanoparticles (AgNPs) have received great attention in the biomedical field because of their intrinsic therapeutic properties. Nanoparticles synthesized from silver have been studied as antimicrobial, antiviral and anticancer agents and found particularly an attractive source for the development of a new and advance group of antimicrobial agents. Objectives: In the present study, silver nanoparticles were synthesized from non-pathogenic Bacillus subtilis strain to assess their antimicrobial activity. Methods: Different strains of Bacillus spp. were selected and screened against silver nitrate (AgNO3) toxicity. Finally, B. subtilis strain (FCBP-WB-0174) was selected based on its silver resistant nature, among other strains. Silver nanoparticles were synthesized and optimized from the supernatant of B. subtilis culture at 37°C by the reduction of silver ions using the various molar concentration of AgNO3. The synthesized AgNPs were characterized by UV-Vis spectrophotometry and scanning electron microscopy (SEM). These synthesized AgNPs were used for evaluating antimicrobial activity against four multidrug-resistant bacterial strains. Results: The silver ion reduction was found at a ratio of 1:1 from all the three molar concentrations (1, 2, and 3 mM) of AgNO3. The characterized nanoparticles were found to have a characteristic absorption peak at 426 nm, and the particles were found to have spherical shape under SEM with an average diameter of about 80 ± 0.18 nm, which was also reconfirmed using Zeta Sizer Nano. Prepared Silver nanoparticles have found potential antimicrobial activities against all tested pathogenic, including strains, e.g., Acinetobacter baumannii, Pseudomonas aeruginosa, Methicillin-Resistant Staphylococcus aureus(MRSA) and Escherichia coli. Conclusions: Effective AgNP’s were produced from selected B. subtilis strain, and the strain itself was resistant to AgNO3. The current study evidenced that biologically synthesized silver nanoparticles from B. subtilis has promising antimicrobial activities against pathogenic and multidrug-resistant bacteria.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes huge economic loss to China's swine industry. Currently, a novel type 2 PRRSV, called the NADC30-like strain, is epidemic in numerous provinces of China. In this study, a NADC30-Like PRRSV strain was isolated in primary alveolar macrophage (PAM) cells from fecal samples collected from a local pig farm, which suffered severe diarrhea. A pathogenicity comparison study was conducted in 6-week-old piglets by inoculating highly pathogenic HP-PRRSV and NADC30-Like PRRSV isolates. RT-qPCR revealed detection of NADC30-Like PRRSV but not the HP-PRRSV in the intestine. PRRSV infection-related lesions were observed in the intestine were further confirmed by histopathological and immunohistochemical examination (IHC). In addition, severe virus infections were also detected by RT-qPCR. Based on clinical observation and pathogenicity experiments, we confirmed that NADC30-Like PRRSV gained more tissue tropism, especially in the small intestine. This may be the one reason explaining why NADC-Like 30 PRRSV become a major epidemic strain in China since the first outbreak in 2013.

Cancer is a leading cause of high death rate worldwide. One strategy to control the disease is the early diagnosis by novel biomarkers that express during early stage of the disease. The recent diagnostic strategies in cancer don’t have enough specificity to promote the detection of cancer at its beginning. Many biomarkers like protein biomarkers and metabolites are being used for diagnosis of various cancer types but miRNAs are excellent among them, because they have distinctive biochemical characteristics. Moreover, to raise the precision and capability of miRNA to diagnose cancer, the analyzing of both miRNAs and as well as selective mRNA will help in creating a more complete categorizer. Virus constitutes the cause of 20% of entire human cancer cases and both RNA and DNA viruses are linked with tumors in both animal and man. Even though many viruses can cause different tumors in animals, only some of them are linked with human cancers and are presently regarded as oncogenic viruses. These viruses include Human Papillomavirus (HPV), Hepatitis B (HBV) and Hepatitis C Virus (HCV), Epstein Barr Virus (EBV), Human Herpes Virus 8 (HHV8), Human T cell Leukemia Virus (HTLV) and Merkel Cell Polyomavirus (MCPyV). Expression data of miRNA in several cancers reveal that miRNA profile is different in cancer cells as compared to normal cells. So, miRNA could be useful biomarker for the detection of cancer. The present study strengthens a foundation and gives a logic to investigate the ability of miRNAs as circulating biomarkers in various cancers.

Complete S1 gene of the Infectious Bronchitis Virus (IBV) was amplified and cloned into transfer vector. Truncated S1 gene designated as Sf200 (containing five antigenic sites located at 24–61, 291–398 and 497–543 amino acid residues of S1 glycoprotein) were amplified by overlap PCR, cloned into prokaryotic expression vector resulting pET-Sf200 and confirmed the construct by sequencing. The recombinant plasmid was identified by restriction enzyme and sequencing analysis. The in vitro expression of the truncated protein was analyzed in E. coli with a molecular weight of 38kDa determined through SDSPAGE and confirmed by Western blotting. The recombinant truncated protein was then purified from the culture media. The immunogenicity of the protein was studied in an animal experiment on mice, in which mice were injected subcutaneously. These findings suggest that the truncated Sf200 expressed in the pET- 32a (+) prokaryotic vector can be used as antigen to detect antibodies against IBV.

Boer crossbred goats (n=48) single born kids, seven-day-old (live weight 4.4±0.09kg) were divided into 3 treatment groups, each having 8 females and 8 males. The kids from first treatment group had free access to creep feed with roughage (CFR), second group had free access to creep feed without roughage (CF) while last group (control) had access to their does’ milk (DM) only. All kids had access to their dam’s milk. The kids were weaned at day 84 and two kids from each treatment group were sacrificed. The results showed that the rumen morphological characteristics, papillary surface area and keratinized layer of CFR (4.15mm2, 50.54µm), were not significantly different from CF (3.81mm2, 49.26µm) (P>0.05) but it was significantly different from kids in DM group (1.17mm2, 26.36µm) (P 0.05), while total surface area of rumen of both creep fed groups was significantly greater than DM group. The rumen tissue weight of creep-fed kids was 17% greater than those of milk fed kids. Overall, the present study showed that the creep feed supplementation to nursing goat kids improve rumen morphology. Read more at http://researcherslinks.com/current-issues/Creep-Feeding-Supplemented-with-Roughages/20/1/1134/html#lyAKjwLAfprvXT3I.99.

Quantum Dots have shown remarkable potentials in biomedical research. Herein, we reported the effects of CdTe quantum dots (QDs) and CdTe@SiO2 nanoparticles (NPs) on human embryonic kidney 293 (HEK 293A) cells with the aim of investigating their in vitro cytotoxicity. The CdTe@SiO2 particles were prepared by reverse microemulsion method. The structural morphology of the CdTe and hydrophilic silica-coated CdTe particles were characterized by transmission electron microscopy (TEM), ultraviolet-visible (UV-vis) spectrometry and photoluminescence (PL) spectrometry. The in vitro cytotoxicity of CdTe QDs and CdTe@SiO2 nanoparticles was assessed in 293A cells using standard MTT assay, western blot and fluorescent microscopy. The results showed that the CdTe and CdTe@SiO2 particles were relatively uniform with the diameter of about 3.8 nm, 75 nm respectively. The cell viability and the adhesion ability were similar to the control 293A cells. The level of the fibronectin protein expression was decreased with the increasing concentration of CdTe while the no effects were observed on expression of beta-actin in CdTe as well as CdTe@SiO2 treated cells even at highest concentration of 45 microg/mL which demonstrated their good biocompatibility to 293A cells. The results indicate that the CdTe@SiO2 nanoparticles are attractive candidates for biological imaging studies as expected.

Encephalomyocarditis virus (EMCV) could infect many host species and cause acute myocarditis and sudden death in pre-weaned piglets. It was necessary to develop new antiviral strategies for the treatment of the virus infection. Here, four plasmids expressing shRNA (small hairpin RNA) targeted to 1D or 3AB protein genes of EMCV were constructed and their inhibition efficiency on the replication of EMCV was evaluated in both BHK21 cells and mice. The results showed that three out of those four shRNA constructs could significantly inhibit EMCV replication in BHK21 cells on the levels of viral RNA and protein. Moreover, it was found that the shRNAs could suppress significantly the load of EMCV in the brain tissue of the mice pretreated with the constructs for 6-24 h. The clinical signs and pathological lesions of the mice in the groups inoculated with the shRNA constructed were milder obviously, compared with those in pSUPER-mN3 and challenge control groups. The survival rates of mice inoculated with pSUPER-3AB-1, pSUPER-3AB-2, and pSUPER-1D-1 for 12 h was 100, 80, and 40%, respectively, while, in the control groups it was only 20%. It indicated that the vector-based shRNA targeting to 3AB and 1D genes might be a potential anti-EMCV strategy.

Infectious bronchitis virus (IBV) causes tremendous economic losses associated with production inefficiencies and mortality in poultry industry worldwide. In the present report, the recombinant adenoviruses expressing chicken granulocyte-macrophage colony stimulating factor (GM-CSF) and S1 gene of nephropathogenic IBV were constructed and characterized. Then, the immunological efficacy and protection against homologous IBV challenge were assessed in specific pathogen free (SPF) chickens. The results showed that the chickens vaccinated in ovo with rAd-S1, rAd-GM-S1 (GM-CSF fused with S1 using glycine linkers) and rAd-GM-CSF plus rAd-S1 (co-administered) developed specific anti-IBV HI antibodies. Moreover, the fusion of the GM-CSF markedly increased spleen cell proliferation and IFN-γ production while mild increased in IL-4 production, which demonstrated the enhancement of cell-mediated immune responses. Following challenge with IBV, the chickens in the group vaccinated with rAd-S1 fused or co-administered with GM-CSF had fewer nephropathic lesions and showed 100% protection as compared to that of rAd-S1 alone which showed 70% protection. It indicated that the single dose in ovo vaccination of the GM-CSF fused or co-administered with S1 of IBV could enhance significantly the humoral, cellular immune responses and provide complete protection against nephropathogenic IBV challenge. This finding may provide basic information for effective in ovo vaccines design against IBV.

Mitigating methane losses from cattle has economic as well as environmental benefits. The aim of this paper is to review the current approaches in relation to associated advantages and disadvantages and future options to reduce enteric methane emission from cattle. Current technologies can be broadly grouped into those that increase productivity of the animal (improved nutrition strategies) so that less methane is produced per unit of meat or milk, and those that directly modify the rumen fermentation so that less methane is produced in total. Data suggest that many of these practices are not appropriate for long term mitigation of methane emissions in ruminants because of their constraints. So it is necessity to develop long term strategies in suppressing methane production. An integrated research investigating animal, plant, microbe and nutrient level strategies would offer a long term solution of methane production. Genetic selection of animals, vaccination, probiotics, prebiotics and plant improvement are the most promising options of all the future approaches discussed. These approaches will reduce enteric methane production without any hazard to animal or environment.

The circulating miRNAs are sufficiently stable and detectable to serve as clinical biomarkers as recent studies have revealed that the aberrant expression of circulating miRNAs can directly reflect disease status. Based on the analysis of the data (using miRanda software, TargetScan software and SOLID high-throughput sequencing) obtained from the literature, Schizophrenia Gene database, NCBI database, the quantification of the nine miRNAs in the serum samples of 115 patients suffering from schizophrenia and 40 healthy individuals using qRT-PCR and semi-nested qRT-PCR was conducted. The results suggested that the miR-181b, miR-219-2-3p, miR-346, miR-195, miR-1308, miR-92a, miR-17, miR-103 and let-7g are the key players to reflect the schizophrenia illnesses status and may serve as candidate biomarkers for diagnosis of schizophrenia. In addition, we also found that the risperidone improved the serum miR-346 level of schizophrenia significantly, and therefore may not be an effective drug in regulating serum miR-346 level of schizophrenia. Furthermore, the expression level of serum miRNAs levels and schizophrenia patients were regardless of family history subtypes, ages, and gender. Collectively, these findings suggested that the serum miRNAs have strong potential to reflect schizophrenia disease status. To the best of our knowledge, this is the first report demonstrating the analysis of the circulating miRNAs in schizophrenia.

Porcine reproductive and respiratory syndrome virus (PRRSV) has recently caused catastrophic losses in swine industry worldwide. Current vaccination strategies only provide a limited protection against PRRSV infection. This study was aimed to construct the recombinant adenovirus co-expressing GP3 and GP5 of highly pathogenic PRRSV fused with swine granulocyte-macrophage colony stimulating factor (GM-CSF) (rAd-GF35), and to detect the immune response in mice and pigs. The results showed that the rAd-GF35 could induce significantly higher PRRSV-specific neutralizing antibodies than the recombinant adenovirus only expressing GP3 and GP5 (rAd-GP35). Moreover, the fusion of GM-CSF markedly increased the secretion of IFN-gamma and IL-4 in PRRSV-stimulated mice lymphocytes culture and pigs sera. Following challenge with PRRSV, piglets inoculated with recombinant rAd-GF35 had lighter clinical signs, lower viremia and less gross lesion of lungs, as compared to that of rAd-GP35 immunized group. It demonstrated that GM-CSF fused with GP3 and GP5 of PRRSV could significantly enhance the humoral and cellular immune responses and provide protection against PRRSV challenge in pigs. The recombinant adenovirus rAd-GF35 might be an attractive candidate vaccine for the prevention and control of highly pathogenic PRRSV infection.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been mainly responsible for the heavy economic losses in pig industry all over the world. Current vaccination strategies provide only a limited protection. In this study recombinant adenoviruses expressing GP3/GP5 of highly pathogenic PRRSV and heat shock protein 70 (HSP70) gene of Heamophilus parasuis were constructed, and the immune responses and protective efficacy against homologous challenge were examined in pigs. The results showed that all animals vaccinated with rAd-GP35 (co-expressing GP3-GP5), rAd-HS35 and rAd-HSA35 (co-expressing GP3-GP5 fused with HSP70 using different linkers), developed specific anti-PRRSV ELISA antibody and neutralizing antibody. The humoral immune responses of rAd-HS35, especially rAd-HSA35 containing 2A of FMDV between HSP70 and GP3 gene, were significantly higher than that of rAd-GP35. Moreover, the fusion of HSP70 markedly induced both IFN-gamma and IL-4 in pigs' sera. Following challenge with PRRSV, pigs inoculated with recombinant rAd-HS35 and rAd-HSA35 showed lighter clinical signs, lower viremia and less pathological lesion of lungs, as compared to those in rAd-GP35 group. Moreover, the protective efficiency induced by rAd-HSA35 was higher than that of rAd-HS35. It indicated that HSP70 fused with GP3 and GP5 of PRRSV could induce enhanced immune responses and provide protection against virulent PRRSV challenge in pigs. The recombinant adenovirus rAd-HSA35 might be an attractive candidate vaccine for the prevention and control of highly pathogenic PRRSV infections.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been mainly responsible for the heavy economic losses in many swine-producing regions. Current vaccination strategies and antiviral drugs provide only limited protection. Consequently, there is a need to develop a new antiviral strategy. In this study, two recombinant adenoviruses expressing short-hairpin RNAs (shRNAs) directed against ORF1b of PRRSV S1 strain were constructed and the inhibition of PRRSV replication was determined. The results showed that pretreatment with these shRNAs delivered by recombinant adenovirus could induce a significant inhibition of viral RNA and protein level in Marc-145 cells infected with PRRSV S1 strains. One recombinant adenovirus (rAd-P2) was found to be also effective in inhibiting the replication of highly virulent PRRSV SY0608 strain in Marc-145 cells and porcine alveolar macrophages at both the protein and ORF1b mRNA level. The antiviral effect was dose-dependent and sustained for at least 96 h. Twenty 6-week old piglets were assigned to four groups each with five piglets. Groups 1 and 2 were inoculated intramuscularly with rAd-P2 and mock construct rAd-mP2 individually. After 24 h, groups 1, 2 and 3 were challenged intramuscularly with the SY0608 strain. Group 4 remained unchallenged but with PBS as mock. The results showed that the viral load of PRRSV in serum and lung tissue of swine was suppressed effectively by rAd-P2. The clinical signs and pathological lesions in the pigs inoculated with rAd-P2 were milder than those in rAd-mP2 negative and PRRSV control. These results indicated that shRNAs mediated by the adenovirus could inhibit PRRSV infection sufficiently in vitro as well as in vivo . RNAi mediated by recombinant adenovirus might be a potential new tool for controlling PRRSV infection. Of course, the protective efficiency of rAd-P2 should be made by using a large number of pigs in future.

Porcine reproductive and respiratory syndrome virus has caused hundreds of thousands of deaths in pig farms in many swine-producing areas in the world in recent years. However, at present there is no effective method to prevent and control the disease, and there is a need to develop new antiviral strategies. In this study, four recombinant adenoviruses expressing shRNAs targeting ORF1b, ORF5 and ORF7 were constructed, and it was found that they could down-regulate effectively specific gene expression and inhibit viral replication in MARC-145 cells when compared to the controls. They could also inhibit effectively PRRSV replication in porcine alveolar macrophages. The inhibition effect was dose-dependent and could be sustained for at least 96h in macrophages. In addition, PRRSV replication could be suppressed significantly by shRNA in cells infected previously or simultaneously with PRRSV. The results indicated that the shRNA-expressing rAd5 targeting to various gene regions of PRRSV might be a potential anti-PRRSV strategy.

Introduction Granulocyte-monocyte colony stimulating factor (GM-CSF) has been reported to to mediate antitumor effects through stimulation of various antigen presentation cells. Clearly, these properties make GM-CSF a potent adjuvant. Therefore, an oncolytic adenovirus coding for GMCSF was engineered. Objective Contraction, identification and characterization of recombinant adenovirus encoding GM-CSF. Methods In this study, a recombinant adenovirus expressing GM-CSF (rAd-GM) was constructed and characterized. Homologous recombination in bacteria followed by transfection of recombinant plasmid into a mammalian packaging cell line, viral production was conveniently followed by the gene incorporated into the viral backbone. Results & Discussion We also showed that the construct produced a 12 kDa recombinant protein which stimulated colony formation in bone marrow cells in vitro , indicating that the recombinant GM-CSF was biologically active. Conclusion Future work will demonstrate the rAd-GM mediated antitumor immunity in rats with experimentally induced breast cancer.

Background H9N2 influenza A viruses have undergone extensive reassortments in different host species, and could lead to the epidemics or pandemics with the potential emergence of novel viruses. Methods To understand the genetic and pathogenic features of early and current circulating H9N2 viruses, 15 representative H9N2 viruses isolated from diseased chickens in northern China between 1998 and 2010 were characterized and compared with all Chinese H9N2 viruses available in the NCBI database. Then, the representative viruses of different genotypes were selected to study the pathogenicity in mice with the aim to investigate the adaptation and the potential pathogenicity of the novel H9N2 reassortants to mammals. Results Our results demonstrated that most of the 15 isolates were reassortants and generated four novel genotypes (B62-B65), which incorporated the gene segments from Eurasian H9N2 lineage, North American H9N2 branch, and H5N1 viruses. It was noteworthy that the newly identified genotype B65 has been prevalent in China since 2007, and more importantly, different H9N2 influenza viruses displayed a diverse pathogenicity to mice. The isolates of the 2008-2010 epidemic (genotypes B55 and B65) were lowly infectious, while two representative viruses of genotypes B0 and G2 isolated from the late 1990s were highly pathogenic to mice. In addition, Ck/SD/LY-1/08 (genotype 63, containing H5N1-like NP and PA genes) was able to replicate well in mouse lungs with high virus titers but caused mild clinical signs. Conclusion Several lines of evidence indicated that the H9N2 influenza viruses constantly change their genetics and pathogenicity. Thus, the genetic evolution of H9N2 viruses and their pathogenicity to mammals should be closely monitored to prevent the emergence of novel pandemic viruses.

Background Encephalomyocarditis virus (EMCV) can cause myocarditis, respiratory failure, reproductive failure, and sudden death in pre-weaned piglets, which has been isolated in China. EMCV VP1 protein was one of the most important structural proteins and played an important role in the protective immunity. In this study, 10 monoclonal antibodies (McAbs) against EMCV VP1 were screened and identified. Results Epitope mapping results indicated that McAbs (6E11, 7A7, 7C9) specifically recognized the linear epitopes V(2)ENAEK(7), McAbs (1D1, 2A2, 5A1, 5A11, 5G1) recognized the epitope F(19)VAQPVY(25), and McAbs 1G8 and 3A9 recognized P(42)IGAFTVK(49). Protein sequence alignment of VP1 with 16 EMCV isolates indicated that the epitope F(19)VAQPVY(25) was conserved in all the reference strains. The epitopes P(42)IGAFTVK(49) and V(2)ENAEK(7) only had 1 or 2 variable amino acid among the reference strains. The 3D model analysis results showed that these epitopes presented as spheres were shown within the context of the complete particle. Conclusions In this study, ten McAbs against EMCV VP1 were developed and three B-cells epitopes (2-7aa, 19-25aa and 42-49aa) were defined in VP1. All the results herein will promote the future investigations into the function of VP1 of EMCV and development of diagnostic methods of EMCV.