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1.  A Single Point Mutation Creating a Furin Cleavage Site in the Spike Protein Renders Porcine Epidemic Diarrhea Coronavirus Trypsin Independent for Cell Entry and Fusion 
Journal of Virology  2015;89(15):8077-8081.
The emerging porcine epidemic diarrhea virus (PEDV) requires trypsin supplementation to activate its S protein for membrane fusion and virus propagation in cell culture. By substitution of a single amino acid in the S protein, we created a recombinant PEDV with an artificial furin protease cleavage site N terminal of the putative fusion peptide (PEDV-SFCS). PEDV-SFCS exhibited trypsin-independent cell-cell fusion and was able to replicate in culture cells independently of trypsin, though to low titer.
doi:10.1128/JVI.00356-15
PMCID: PMC4505673  PMID: 25972540
2.  Inhibitory effects of metformin at low concentration on epithelial–mesenchymal transition of CD44+CD117+ ovarian cancer stem cells 
Background
Although metformin, a first-line drug for treating diabetes, may play an important role in inhibition of epithelial ovarian cancer cell growth and cancer stem cells (CSCs), metformin at low dose showed less effect on the proliferation of ovarian cancer cells. In this study, we evaluated the effect of metformin at low dose on ovarian CSCs in order to understand the molecular mechanisms underlying.
Methods
The inhibitory effects of metformin at los dose on proliferation and population of ovarian cancer cells including SKOV3 and A2780 were assessed by cell proliferation assay and flow cytometry. Quantitative real-time PCR assay on expression of Bcl-2, Survivin and Bax was performed to determine the effect of metformin at low dose on epithelial-mesenchymal transition (EMT) of cancer cells and CSCs. Tumor sphere formation assay was also performed to evaluate the effect of metformin on spheres forming ability of CSCs. The therapeutic efficacy and the anti-CSC effects of metformin at low dose were investigated by using both SKOV3 cells and primary tumor xenografts. In addition, the CSC frequency and EMT in tumor xenograft models were also assessed by flow cytometry and quantitative real-time PCR.
Results
Metformin at low dose did not affect the proliferation of ovarian cancer cells. However, it inhibited population of CD44+CD117+ selectively, neither CD133+ nor ALDH+ cells. It suppressed expression of snail2, twist and vimentin significantly in cancer cells and CD44+CD117+ CSCs in vitro. Low dose of metformin reduced survivin expression in CSCs. Low concentrations of metformin inhibited the secondary and the tertiary tumor sphere formation, decreased SKOV3 and primary ovarian tumor xenograft growth, enhanced the anticancer effect of cisplatin, and lowered the proportion of CD44+CD117+ CSCs in the xenograft tissue. Metformin was also associated with a reduction of snail2, twist, and vimentin in CD44+CD117+ ovarian CSCs in vivo.
Conclusions
Our results implicate that metformin at low dose inhibits selectively CD44+CD117+ ovarian CSCs through inhibition of EMT and potentiates the effect of cisplatin.
doi:10.1186/s13287-015-0249-0
PMCID: PMC4697317  PMID: 26718286
Metformin; Epithelial–mesenchymal transition; Ovarian cancer stem cells
3.  Gamma knife radiosurgery (GKRS) for pineal region tumors: a study of 147 cases 
Background
The purpose of this study is to evaluate the effectiveness of gamma knife radiosurgery (GKRS) in the treatment of pineal region tumors (PRTs).
Methods
We retrospectively reviewed 147 cases of PRTs primarily treated with GKRS at our hospital between 1999 and 2009. Mean follow-up time was 67 months (range 60.5–100.1). The local tumor control rates (LTCRs) and overall survival rates were calculated to evaluate the results of the GKRS treatment.
Results
At 2 months after GKRS, tumor volume was significantly reduced in 91 cases (61.9 %). At 6 months, average tumor volume was 4.2 cm3 as compared to 8.47 cm3 before GKRS. By 1 year after GKRS, the tumor completely disappeared in 57 patients. Fourteen patients underwent second treatment, and one patient had third treatment. The overall survival rates were 72.1 % at 3 years and 66.7 % at 5 years for all patients and 62.4 % at 3 years and 54.5 % at 5 years for germ cell tumors (GCTs). The LTCRs were 94.30 % at 3 years and 90.80 % at 5 years for all patients and 88.00 % at 3 years and 77.27 % at 5 years for GCTs.
Conclusions
GKRS is an effective and safe modality that can be widely used to PRTs as the primary therapy.
doi:10.1186/s12957-015-0720-5
PMCID: PMC4617952  PMID: 26490154
Gamma knife radiosurgery; Pineal region tumors; Germ cell tumors; Follow-up
4.  Radiofrequency ablation of advanced lung tumors: imaging features, local control, and follow-up protocol 
To prospectively observe imaging features and local control of advanced lung tumors after radiofrequency ablation (RFA), and to propose a follow-up protocol post-ablation. 58 stage IV malignant lung tumor patients were enrolled in our study. One hundred of lung lesions were performed 77 sessions of RFA. Enhanced computed tomographic (CT) images of pre-ablation, 1-month, and 3-month post-ablation and thereafter every 3 months were obtained. Positron emission tomographic/CT (PET/CT) was performed pre-ablation, 3-month post-ablation and thereafter every 6 months. The CT size, shape, enhancement, and PET/CT metabolic activity of the ablated zone were analyzed to assess local lesion control. There was significant difference in lesion size between pre-ablation and 1-month post-ablation (P=0.000), 1 and 3-month post-ablation (P=0.000), 3 and 6-month post-ablation (P=0.006). Metabolic activity of the ablated zone after 3 months decreased markedly as compared with pre-ablation (p=0.001). Local control rate was 88%, and forms of definite recurrence or residual included increased size, nodular enhancement, and central enhancement. Time to local progression (TTLP), progression-free survival (PFS), and overall survival (OS) were 15.4±7.5, 9.6±5.8 and 18.0±7.0 months respectively. No death related to operation occurred, and the main complication rate was 29%, of which 9% needed clinical management. RFA is a safe and effective approach for local control of lung tumors even if in advanced patients. To obtain definite CT evaluation, lesion size at 1-month post-ablation as the baseline is appropriate, with efficacy assessment 6-month post-ablation. PET/CT is a useful tool to predict recurrence or residual at least 3 months post-ablation.
PMCID: PMC4694311  PMID: 26770411
Radiofrequency ablation; advanced lung tumors; imaging features; local control; follow-up protocol
5.  Evaluation of anti-smoking television advertising on tobacco control among urban community population in Chongqing, China 
Tobacco Induced Diseases  2015;13(1):31.
Background
China is the largest producer and consumer of tobacco in the world. Considering the constantly growing urban proportion, persuasive tobacco control measures are important in urban communities. Television, as one of the most pervasive mass media, can be used for this purpose.
Methods
The anti-smoking advertisement was carried out in five different time slots per day from 15 May to 15 June in 2011 across 12 channels of Chongqing TV. A cross-sectional study was conducted in the main municipal areas of Chongqing. A questionnaire was administered in late June to 1,342 native residents aged 18–45, who were selected via street intercept survey.
Results
Respondents who recognized the advertisement (32.77 %) were more likely to know or believe that smoking cigarettes caused impotence than those who did not recognize the advertisement (26.11 %). According to 25.5 % of smokers, the anti-smoking TV advertising made them consider quitting smoking. However, females (51.7 %) were less likely to be affected by the advertisement to stop and think about quitting smoking compared to males (65.6 %) (OR = 0.517, 95 % CI [0.281–0.950]). In addition, respondents aged 26–35 years (67.4 %) were more likely to try to persuade others to quit smoking than those aged 18–25 years (36.3 %) (OR = 0.457, 95 % CI [0.215–0.974]). Furthermore, non-smokers (87.4 %) were more likely to find the advertisement relevant than smokers (74.8 %) (OR = 2.34, 95 % CI [1.19–4.61]).
Conclusions
This study showed that this advertisement did not show significant differences on smoking-related knowledge and attitude between non-smokers who had seen the ad and those who had not. Thus, this form may not be the right tool to facilitate change in non-smokers. The ad should instead be focused on the smoking population. Gender, smoking status, and age influenced the effect of anti-smoking TV advertising on the general population in China.
doi:10.1186/s12971-015-0057-4
PMCID: PMC4558728  PMID: 26339225
Anti-smoking; television advertising; evaluation; urban community; China
6.  Large-mode-area single-mode-output Neodymium-doped silicate glass all-solid photonic crystal fiber 
Scientific Reports  2015;5:12547.
We have demonstrated a 45 μm core diameter Neodymium-doped all-solid silicate glass photonic crystal fiber laser with a single mode laser output. The structure parameters and modes information of the fiber are both demonstrated by theoretical calculations using Finite Difference Time Domain (FDTD) method and experimental measurements. Maximum 0.8 W output power limited by launched pump power has been generated in 1064 nm with laser beam quality factor M2 1.18.
doi:10.1038/srep12547
PMCID: PMC4648479  PMID: 26205850
7.  Homoepitaxial SrTiO3(111) Film with High Dielectric Performance and Atomically Well-Defined Surface 
Scientific Reports  2015;5:10634.
The six-fold symmetry possessed by the (111) surfaces of perovskite oxides allows the epitaxial growth of novel quantum materials such as topological insulators. The dielectric SrTiO3(111) thin film is an ideal buffer layer, providing the readily tunability of charge density in gate-controlled structures. But the high-quality film growth is challenging due to its strong surface polarity as well as the difficulty of obtaining the chemical stoichiometry. Here we show that the layer-by-layer growth of homoepitaxial SrTiO3(111) thin films can be achieved in molecular beam epitaxy method by keeping the growing surface reconstructed. And the cation stoichiometry is optimized precisely with the reflective high energy electron diffraction as the feedback signal that changes sensitively to the variation of metal concentration during growth. With atomically well-defined surfaces, the SrTiO3(111) films show high dielectric performance with the charge density modulated in the range of 2 × 1013/cm2 with the back gate voltage lower than 0.2 V. Methods of further broadening the range are also discussed.
doi:10.1038/srep10634
PMCID: PMC4466594  PMID: 26073468
8.  Clinical features and mutation status of EGFR, KRAS, BRAF, EML4-ALK and ROS1 between surgical resection samples and non surgical resection samples in lung cancer 
Journal of Thoracic Disease  2015;7(5):875-880.
Background
Target therapy is the first-line treatment in lung cancer. The testing of driver gene mutations is crucial for decision of treatment. Many lung cancer patients are in advanced grade, and lose the chance of operation.
Methods
The tissue used to perform mutation testing is only from biopsy. In order to analysis the difference between surgical resection samples (SRSs) and non-surgical resection samples (NSRSs), 1,357 surgical tissues and 145 biopsy samples histopathologically diagnosed with lung cancer were collected to detect the mutation status of EGFR, KRAS, BRAF, EML4-ALK and ROS1 in this study.
Results
There were no significant differences of age, gender, and histological type between the two group patients we collected; however, the significant difference was present in grade. More early stage patients were in the surgical group, but more advanced stage lung cancer patients were in non surgical group. In the mutation analysis, we also found no significant differences in all driver genes we detected between the two groups.
Conclusions
Both surgical resection samples and biopsy samples could be used to perform the testing the driver gene mutation.
doi:10.3978/j.issn.2072-1439.2015.04.49
PMCID: PMC4454852  PMID: 26101643
Surgical resection sample (SRS); non-surgical resection sample (NSRS); lung cancer; driver gene mutation
9.  Cloning and Characterisation of Two H+ Translocating Organic Pyrophos-phatase Genes in Salix and Their Expression Differences in Two Willow Varieties with Different Salt Tolerances 
Current Genomics  2014;15(5):341-348.
Willows are one of the most important tree species for landscaping, biofuel and raw timber. Screening salt-tolerant willow varieties is an effective approach to balance wood supply and demand. However, more salt-tolerant willow varieties are required and little is known regarding the mechanism of salt tolerance at the gene expression level. In this paper, two willow varieties were studies in terms of their differences in salt-tolerances and mechanism of salt tolerance at the level of VP1 gene expression. The results showed that Salix L0911 (L0911) had higher biomass than Salix matsudana (SM), and salt injuries were less severe in L0911 than in SM. The activities of peroxidase and superoxide dismutase, as well as the contents of soluble protein and proline, were higher in L0911 than in SM, whereas the contents of Na+ and K+, as well as the Na+/K+ ratio, were lower in L0911 than in SM. Two VP1 genes (VP1.1 and VP1.2) cloned in L0911 and SM had similar sequences and structures. VP1.1 and VP1.2 belonged to different subgroups. Total expression levels of the VP1.1 gene in both roots and leaves of L0911 were higher than that in SM under normal conditions. Under salt stress, expression of VP1 in SM roots initially increased and then decreased, whereas the expression of VP1 in leaves of L0911 and SM, as well as in roots of L0911, decreased with increasing salt concentrations. This study increased our understanding of the salt-tolerance mechanism of willow and may facilitate the selection of salt-tolerant willow resources.
doi:10.2174/138920291505141106102544
PMCID: PMC4245694  PMID: 25435797
Willow; VP1 gene; Salt tolerance; Gene expression; Phylogenetic analysis.
10.  A prospective clinical trial of radiofrequency ablation for pulmonary metastases 
Molecular and Clinical Oncology  2015;3(3):559-562.
Pulmonary oligometastases can be treated by local treatment options, such as resection, radiofrequency ablation (RFA), and radiotherapy, in selected patients. In the present study, 67 patients with 115 pulmonary metastases were treated, and all the lesions were treated by RFA and followed with a computed tomography scan. The local control, overall survival (OS), progression-free survival (PFS) and treatment-related toxicity were observed. Median follow-up duration after the initial RFA was 24 months (range, 3–39 months). The median PFS from RFA was 14 months [95% confidence interval (CI), 11.6–16.4]. The 6-, 12- and 18-month PFS rates were 82.1, 55.7 and 27.5%, respectively. The median OS rate from RFA was 24 months (95% CI, 18.2–29.8). The 1-, 2- and 3-year OS rates were 83.6, 46.3 and 14.3%, respectively. Primary tumor was significantly correlated to PFS and OS on multivariate analysis, and other variates showed no significance. Therefore, RFA is safe for patient treatment and can be considered as a promising treatment option for patients with pulmonary metastases.
doi:10.3892/mco.2015.525
PMCID: PMC4471635  PMID: 26137267
pulmonary metastases; radiofrequency ablation; local tumor control; prognostic factors
11.  Proteolytic Activation of the Porcine Epidemic Diarrhea Coronavirus Spike Fusion Protein by Trypsin in Cell Culture 
Journal of Virology  2014;88(14):7952-7961.
ABSTRACT
Isolation of porcine epidemic diarrhea coronavirus (PEDV) from clinical material in cell culture requires supplementation of trypsin. This may relate to the confinement of PEDV natural infection to the protease-rich small intestine of pigs. Our study focused on the role of protease activity on infection by investigating the spike protein of a PEDV isolate (wtPEDV) using a reverse genetics system based on the trypsin-independent cell culture-adapted strain DR13 (caPEDV). We demonstrate that trypsin acts on the wtPEDV spike protein after receptor binding. We mapped the genetic determinant for trypsin-dependent cell entry to the N-terminal region of the fusion subunit of this class I fusion protein, revealing a conserved arginine just upstream of the putative fusion peptide as the potential cleavage site. Whereas coronaviruses are typically processed by endogenous proteases of the producer or target cell, PEDV S protein activation strictly required supplementation of a protease, enabling us to study mechanistic details of proteolytic processing.
IMPORTANCE Recurring PEDV epidemics constitute a serious animal health threat and an economic burden, particularly in Asia but, as of recently, also on the North-American subcontinent. Understanding the biology of PEDV is critical for combatting the infection. Here, we provide new insight into the protease-dependent cell entry of PEDV.
doi:10.1128/JVI.00297-14
PMCID: PMC4097775  PMID: 24807723
12.  Transcription bypass of DNA lesions enhances cell survival but attenuates transcription coupled DNA repair 
Nucleic Acids Research  2014;42(21):13242-13253.
Transcription-coupled DNA repair (TCR) is a subpathway of nucleotide excision repair (NER) dedicated to rapid removal of DNA lesions in the transcribed strand of actively transcribed genes. The precise nature of the TCR signal and how the repair machinery gains access to lesions imbedded in stalled RNA polymerase II (RNAP II) complexes in eukaryotic cells are still enigmatic. RNAP II has an intrinsic capacity for transcription bypass of DNA lesions by incorporation or misincorporation of nucleotides across the lesions. It has been suggested that transcription bypass of lesions, which exposes the lesions, may be required for TCR. Here, we show that E1103G mutation of Rpb1, the largest subunit of RNAP II, which promotes transcription bypass of UV-induced cyclobutane pyrimidine dimers (CPDs), increases survival of UV irradiated yeast cells but attenuates TCR. The increased cell survival is independent of any NER subpathways. In contrast, G730D mutation of Rpb1, which impairs transcription bypass of CPDs, enhances TCR. Our results suggest that transcription bypass of lesions attenuates TCR but enhances cell tolerance to DNA lesions. Efficient stalling of RNAP II is essential for efficient TCR.
doi:10.1093/nar/gku1150
PMCID: PMC4245964  PMID: 25389266
13.  Clinical efficacy of radical nephrectomy versus nephron-sparing surgery on localized renal cell carcinoma 
Background
The aim of the present study was to compare the clinical efficacy of radical nephrectomy (RN) with nephron-sparing surgery (NSS) in treating patients with localized renal cell carcinoma (RCC).
Methods
The literature search was performed in PubMed, MEDLINE Springer, Elsevier Science Direct, Cochrane Library, and Google Scholar up to December 2012. The software Review Manager 5.1 and the STATA software package v.11.0 were used for analyses. The odds ratios (ORs) and its 95% confidence interval (95% CI) were calculated for comparison. Subgroup analyses were performed based on the tumor size of RCC.
Results
In total, 10 studies with 10,174 RCC patients (7,050 treated with RN and 3,124 treated with NSS) were selected. The pooled estimate (OR = 1.58, 95% CI = 1.15–2.15, P = 0.004) showed a significantly lower rate of cancer-specific deaths in the patients treated with NSS compared to RN. However, no statistically significant differences were found in the rate of tumor recurrence (OR = 0.84, 95% CI = 0.67–1.06, P = 0.14) and complications (OR = 0.91, 95% CI = 0.51–1.63, P = 0.74) between the patients treated with NSS and RN. In addition, all the subgroup analyses presented consistent results with the overall analyses.
Conclusions
NSS had no significantly different from RN in tumor recurrence and complications for localized RCC. However, the significantly lower rate of cancer-specific deaths supported the use of NSS not only for RCC with tumor size >4.0 cm but also for tumor sizes ≤4.0 cm compared with RN.
doi:10.1186/s40001-014-0058-4
PMCID: PMC4226856  PMID: 25374003
Meta-analysis; Nephron-sparing surgery; Radical nephrectomy; Renal cell carcinoma
14.  Further Study of Multigranulation T-Fuzzy Rough Sets 
The Scientific World Journal  2014;2014:927014.
The optimistic multigranulation T-fuzzy rough set model was established based on multiple granulations under T-fuzzy approximation space by Xu et al., 2012. From the reference, a natural idea is to consider pessimistic multigranulation model in T-fuzzy approximation space. So, in this paper, the main objective is to make further studies according to Xu et al., 2012. The optimistic multigranulation T-fuzzy rough set model is improved deeply by investigating some further properties. And a complete multigranulation T-fuzzy rough set model is constituted by addressing the pessimistic multigranulation T-fuzzy rough set. The full important properties of multigranulation T-fuzzy lower and upper approximation operators are also presented. Moreover, relationships between multigranulation and classical T-fuzzy rough sets have been studied carefully. From the relationships, we can find that the T-fuzzy rough set model is a special instance of the two new types of models. In order to interpret and illustrate optimistic and pessimistic multigranulation T-fuzzy rough set models, a case is considered, which is helpful for applying these theories to practical issues.
doi:10.1155/2014/927014
PMCID: PMC4156983  PMID: 25215336
15.  Insights into how Spt5 functions in transcription elongation and repressing transcription coupled DNA repair 
Nucleic Acids Research  2014;42(11):7069-7083.
Spt5, a transcription elongation factor, and Rpb4, a subunit of RNA polymerase II (RNAP II) that forms a subcomplex with Rpb7, play important roles in transcription elongation and repression of transcription coupled DNA repair (TCR) in eukaryotic cells. How Spt5 physically interacts with RNAP II, and if and/or how Spt5 and Rpb4/7 coordinate to achieve the distinctive functions have been enigmatic. By site-specific incorporation of the unnatural amino acid p-benzoyl-L-phenylalanine, a photoreactive cross-linker, we mapped interactions between Spt5 and RNAP II in Saccharomyces cerevisiae. Through its KOW4-5 domains, Spt5 extensively interacts with Rpb4/7. Spt5 also interacts with Rpb1 and Rpb2, two largest subunits of RNAP II, at the clamp, protrusion and wall domains. These interactions may lock the clamp to the closed conformation and enclose the DNA being transcribed in the central cleft of RNAP II. Deletion of Spt5 KOW4-5 domains decreases transcription elongation and derepresses TCR. Our findings suggest that Spt5 is a key coordinator for holding the RNAP II complex in a closed conformation that is highly competent for transcription elongation but repressive to TCR.
doi:10.1093/nar/gku333
PMCID: PMC4066765  PMID: 24813444
16.  Broad Activity against Porcine Bacterial Pathogens Displayed by Two Insect Antimicrobial Peptides Moricin and Cecropin B 
Molecules and Cells  2013;35(2):106-114.
In response to infection, insects produce a variety of antimicrobial peptides (AMPs) to kill the invading pathogens. To study their physicochemical properties and bioactivities for clinical and commercial use in the porcine industry, we chemically synthesized the mature peptides Bombyx mori moricin and Hyalophora cecropia cecropin B. In this paper, we described the antimicrobial activity of the two AMPs. Moricin exhibited antimicrobial activity on eight strains tested with minimal inhibitory concentration values (MICs) ranging between 8 and 128 μg/ml, while cecropin B mainly showed antimicrobial activity against the Gramnegative strains with MICs ranging from 0.5 to 16 μg/ml. Compared to the potent antimicrobial activity these two AMPs displayed against most of the bacterial pathogens tested, they exhibited limited hemolytic activity against porcine red blood cells. The activities of moricin and cecropin B against Haemophilus parasuis SH 0165 were studied in further detail. Transmission electron microscopy (TEM) of moricin and cecropin B treated H. parasuis SH 0165 indicated extensive damage to the membranes of the bacteria. Insights into the probable mechanism utilized by moricin and cecropin B to eliminate pathogens are also presented. The observations from this study are important for the future application of AMPs in the porcine industry.
doi:10.1007/s10059-013-2132-0
PMCID: PMC3887904  PMID: 23456332
antimicrobial peptide; cecropin B; Haemophilus parasuis SH 0165; moricin; transmission electron microscopy
17.  Computed tomography-guided core needle biopsy of lung lesions: Diagnostic yield and correlation between factors and complications 
Oncology Letters  2013;7(1):288-294.
The aim of the present study was to determine the diagnostic accuracy of computed tomography (CT)-guided core needle biopsy (CNB) and to retrospectively analyze the correlation between the factors and complications of the procedure. Between January 2009 and June 2010, CNB was performed on 345 lung lesions in 343 patients. These patients were then followed up for at least two years. The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the CNB diagnoses were calculated. The correlation between factors, such as smoking, positoin and maximal diameter, and the complications of pneumothorax and hemorrhage was analyzed by χ2 test. The sensitivity, specificity, accuracy, PPV and NPV of the CNB diagnoses were 97.3, 100, 97.7, 100 and 87.7%, respectively. A statistically significant correlation was found between pneumothorax and the factors of smoking (P=0.015) and position (P<0.01) and length of the needle in the normal parenchyma (P=0.011), as well as between hemorrhage and the maximal diameter (P=0.005) and length of the needle in the normal parenchyma (P<0.01) and the frequency of needle adjustments (P<0.01). A CT-guided core needle biopsy of the lung lesions provides a high diagnostic yield. Smoking, the decubitus position and a longer length of the needle in the normal parenchyma were found to represent risk factors for a pneumothorax. In addition, a small diameter and longer length of the needle in the normal parenchyma and a more frequent adjustment of the needle were poor predictive factors of hemorrhage.
doi:10.3892/ol.2013.1680
PMCID: PMC3861590  PMID: 24348866
CT-guided; core needle biopsy; pneumothorax; hemorrhage; core needle biopsy diagnosis; final diagnosis
18.  Transcription analysis of the porcine alveolar macrophage response to porcine circovirus type 2 
BMC Genomics  2013;14:353.
Background
Porcine circovirus type 2 (PCV2) is the causal agent of postweaning multisystemic wasting syndrome (PMWS), which has severely impacted the swine industry worldwide. PCV2 triggers a weak and atypical innate immune response, but the key genes and mechanisms by which the virus interferes with host innate immunity have not yet been elucidated. In this study, genes that control the response of primary porcine alveolar macrophages (PAMs), the main target of PCV2, were profiled in vitro.
Results
PAMs were successfully infected by PCV2-WH strain, as evidenced quantitative real-time polymerase chain reaction (qPCR) and immunofluorescence assay (IFA) results. Infection-related differential gene expression was investigated using pig microarrays from the US Pig Genome Coordination Program and validated by real-time PCR and enzyme-linked immunosorbent assay (ELISA). Microarray analysis at 24 and 48 hours post-infection (HPI) revealed 266 and 175 unique genes, respectively, that were differentially expressed (false discovery rate <0.05; fold-change >2). Only six genes were differentially expressed between 24 and 48 HPI. The up-regulated genes were principally related to immune response, cytokine activity, locomotion, regulation of cell proliferation, apoptosis, cell growth arrest, and antigen procession and presentation. The down-regulated genes were mainly involved in terpenoid biosynthesis, carbohydrate metabolism, translation, proteasome degradation, signal transducer activity, and ribosomal proteins, which were representative of the reduced vital activity of PCV2-infected cells.
Conclusions
PCV2 infection of PAMs causes up-regulation of genes related to inflammation, indicating that PCV2 may induce systematic inflammation. PCV2 persistently induced cytokines, mainly through the Toll-like receptor (TLR) 1 and TLR9 pathways, which may promote high levels of cytokine secretion. PCV2 may prevent apoptosis in PAMs by up-regulating SERPINB9 expression, possibly to lengthen the duration of PCV2 replication-permissive conditions. The observed gene expression profile may provide insights into the underlying immunological response and pathological changes that occur in pigs following PCV2 infection.
doi:10.1186/1471-2164-14-353
PMCID: PMC3680065  PMID: 23711280
19.  Efficacy of single dose of an inactivated porcine circovirus type 2 (PCV2) whole-virus vaccine with oil adjuvant in piglets 
Background
Post-weaning multisystemic wasting syndrome (PMWS) associated with PCV2 is one of the most costly diseases currently faced by the swine industry. The development of effective vaccines against PCV2 infection has been accepted as an important strategy in the prophylaxis of PMWS.
Methods
In the present study, a PK-15 cell-adapted formalin-inactivated prototype vaccine candidate was prepared using a strain of PCV2 from China. Inactivation of the virus was accomplished using a standard formalin inactivation protocol. The protective properties of the inactivated PCV2 vaccine were evaluated in piglets. Ten 28-day-old pigs were randomly assigned to two groups, each with five. Group 1 was vaccinated intramuscularly with the inactivated virus preparation; Group 2 received sterile PBS as a placebo. By 28 days post-vaccination (DPV), Groups 1 and 2 were challenged intranasally and intramuscularly with 5 × 107 TCID50 of a virulent PCV2 isolate.
Results
The vaccinated pigs seroconverted to PCV2 and had high levels of serum antibodies to PCV2 at 28 days after vaccination, whereas the control pigs remained seronegative. No significant signs of clinical disease were recorded following the challenge with PCV2, but moderate amounts of PCV2 antigen were detected in most lymphoid organs of the control pigs. PCV2 was detected in two out of the five vaccinated pigs. Furthermore, pathological lesions and viremia were milder in the vaccinated group.
Conclusions
The obtained results indicate that the inactivated PCV2 virus vaccine with an oil adjuvant induce an immunological response in pigs that appears to provide protection from infection with PCV2. The vaccine, therefore, may have the potential to serve as a vaccine aimed to protect pigs from developing PMWS.
doi:10.1186/1751-0147-54-67
PMCID: PMC3539982  PMID: 23171757
Porcine circovirus type 2; Post-weaning multisystemic wasting syndrome; Single-dose immunization; Formalin-inactivated vaccine
20.  Design and Synthesis of Benzenesulfonamide Derivatives as Potent Anti-Influenza Hemagglutinin Inhibitors 
ACS Medicinal Chemistry Letters  2011;2(8):603-607.
Structural optimization of salicylamide-based hemagglutinin (HA) inhibitor 1 resulted in the identification of cis-3-(5-hydroxy-1,3,3-trimethylcyclohexylmethylamino)benzenesulfonamide 28 and its derivatives as potent anti-influenza agents. The lead compound 28 and its 2-chloro analogue 40 can effectively prevent cytopathic effects (CPE) caused by infection of influenza A/Weiss/43 strain (H1N1) with EC50 values of 210 and 86 nM, respectively. Mechanism of action studies indicate that 40 and its analogues inhibit the virus fusion with host endosome membrane by binding to HA and stabilizing the prefusion HA structure. With significantly improved metabolic stability, the reported series represents the first generation of orally bioavailable HA inhibitors that have a good selectivity window and potential for further development as novel anti-influenza agents.
doi:10.1021/ml2000627
PMCID: PMC4018120  PMID: 24900355
Anti-influenza; hemagglutinin; benzenesulfonamide; inhibitor
21.  New Variants of Porcine Epidemic Diarrhea Virus, China, 2011 
Emerging Infectious Diseases  2012;18(8):1350-1353.
In 2011, porcine epidemic diarrhea virus (PEDV) infection rates rose substantially in vaccinated swine herds. To determine the distribution profile of PEDV outbreak strains, we sequenced the full-length spike gene from samples from 9 farms where animals exhibited severe diarrhea and mortality rates were high. Three new PEDV variants were identified.
doi:10.3201/eid1808.120002
PMCID: PMC3414035  PMID: 22840964
porcine epidemic diarrhea virus; prevalence; China; phylogeny; variant strain; viruses
22.  Combination of CT-guided hookwire localization and video-assisted thoracoscopic surgery for pulmonary nodular lesions: Analysis of 103 patients 
Oncology Letters  2012;4(4):824-828.
At present, there is no standardized method for the diagnosis and management of pulmonary nodular lesions (PNLs) smaller than 3 cm. This study investigated the use of computed tomography (CT)-guided hookwire localization and video-assisted thoracoscopic surgery (VATS) for PNLs. A total of 103 patients undergoing CT-guided hook-wire localization and VATS were enrolled, and 107 lesions were collected. We assessed the localization achievement ratio, complications rate, conversion thoracotomy rate, intraoperative dislodgement rate, pathological diagnosis rate, duration of surgery and average days of hospitalization. All 107 nodules from 103 patients were successfully localized (100%), the asymptomatic pneumothorax rate was 36.9%, the asymptomatic hemorrhage rate was 40.8% and the simultaneous pneumothorax and hemorrhage rate was 8.7%. A conversion thoracotomy was required in 2 (1.9%) patients and the intraoperative dislodgement rate was 2.9%. The average time for localization was 11±4 min, and the average times for wedge resection and lobectomies were 16±2 and 95±30 min, respectively. The mean hospitalization time following the surgery was 6±3 days. All 107 nodules managed to achieve pathological diagnoses. A combination of CT-guided hook-wire localization and VATS for PNL is a safe and efficient procedure of great clinical value.
doi:10.3892/ol.2012.800
PMCID: PMC3506590  PMID: 23205107
pulmonary nodular lesions; ground-glass nodules; hookwire system; video-assisted thoracoscopic surgery
23.  Transcription analysis on response of porcine alveolar macrophages to Haemophilus parasuis 
BMC Genomics  2012;13:68.
Background
Haemophilus parasuis (H. parasuis) is the etiological agent of Glässer's disease in pigs. Currently, the molecular basis of this infection is largely unknown. The innate immune response is the first line of defense against the infectious disease. Systematical analysis on host innate immune response to the infection is important for understanding the pathogenesis of the infectious microorganisms.
Results
A total of 428 differentially expressed (DE) genes were identified in the porcine alveolar macrophages (PAMs) 6 days after H. parasuis infection. These genes were principally related to inflammatory response, immune response, microtubule polymerization, regulation of transcript and signal transduction. Through the pathway analysis, the significant pathways mainly concerned with cell adhesion molecules, cytokine-cytokine receptor interaction, complement and coagulation cascades, toll-like receptor signaling pathway, MAPK signaling pathway, suggesting that the host took different strategies to activate immune and inflammatory response upon H. parasuis infection. The global interactions network and two subnetworks of the proteins encoded by DE genes were analyzed by using STRING. Further immunostimulation analysis indicated that mRNA levels of S100 calcium-binding protein A4 (S100A4) and S100 calcium-binding protein A6 (S100A6) in porcine PK-15 cells increased within 48 h and were sustained after administration of lipopolysaccharide (LPS) and Poly (I:C) respectively. The s100a4 and s100a6 genes were found to be up-regulated significantly in lungs, spleen and lymph nodes in H. parasuis infected pigs. We firstly cloned and sequenced the porcine coronin1a gene. Phylogenetic analysis showed that poCORONIN 1A belonged to the group containing the Bos taurus sequence. Structural analysis indicated that the poCORONIN 1A contained putative domains of Trp-Asp (WD) repeats signature, Trp-Asp (WD) repeats profile and Trp-Asp (WD) repeats circular profile at the N-terminus.
Conclusions
Our present study is the first one focusing on the response of porcine alveolar macrophages to H. parasuis. Our data demonstrate a series of genes are activated upon H. parasuis infection. The observed gene expression profile could help screening the potential host agents for reducing the prevalence of H. parasuis and further understanding the molecular pathogenesis associated with H. parasuis infection in pigs.
doi:10.1186/1471-2164-13-68
PMCID: PMC3296652  PMID: 22330747
24.  Use of a radiopaque localizer grid to reduce radiation exposure 
Background
Minimally invasive spine surgery requires placement of the skin incision at an ideal location in the patient's back by the surgeon. However, numerous fluoroscopic x-ray images are sometimes required to find the site of entry, thereby exposing patients and Operating Room personnel to additional radiation. To minimize this exposure, a radiopaque localizer grid was devised to increase planning efficiency and reduce radiation exposure.
Results
The radiopaque localizer grid was utilized to plan the point of entry for minimally invasive spine surgery. Use of the grid allowed the surgeon to accurately pinpoint the ideal entry point for the procedure with just one or two fluoroscopic X-ray images.
Conclusions
The reusable localizer grid is a simple and practical device that may be utilized to more efficiently plan an entry site on the skin, thus reducing radiation exposure. This device or a modified version may be utilized for any procedure involving the spine.
doi:10.1186/1750-1164-5-6
PMCID: PMC3177900  PMID: 21827694
Radiation; Exposure; Minimally Invasive; Spine Surgery; Localization; Innovation; Grid

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