Efficacy evaluation of a bivalent subunit vaccine against classical swine fever virus and porcine circovirus type 2.

Classical swine fever virus (CSFV) and porcine circovirus type 2 (PCV2) are two of the most devastating and economically significant pathogens affecting pig populations worldwide. Administration of a combination of vaccines against swine pathogens has been demonstrated to be as efficacious as the administration of single vaccines. In this study, we developed and tested a novel bivalent subunit vaccine against CSFV and PCV2. The safety and efficacy of this vaccine were demonstrated in mice and specific pathogen-free (SPF) piglets. In addition to investigating the serological responses after immunization, challenge studies with both viruses were also conducted. The results showed that this CSFV/PCV2 bivalent vaccine elicited a high level of neutralizing antibodies against both viruses and provided protection in challenge studies. In conclusion, the CSFV/PCV2 bivalent vaccine is safe and effective against CSFV or PCV2 challenge.

Geometry-Based Camera Calibration Using Closed-Form Solution of Principal Line.

Camera calibration is a crucial prerequisite in many applications of computer vision. In this paper, a new geometry-based camera calibration technique is proposed, which resolves two main issues associated with the widely used Zhang's method: (i) the lack of guidelines to avoid outliers in the computation and (ii) the assumption of fixed camera focal length. The proposed approach is based on the closed-form solution of principal lines with their intersection being the principal point while each principal line can concisely represent relative orientation/position (up to one degree of freedom for both) between a special pair of coordinate systems of image plane and calibration pattern. With such analytically tractable image features, computations associated with the calibration are greatly simplified, while the guidelines in (i) can be established intuitively. Experimental results for synthetic and real data show that the proposed approach does compare favorably with Zhang's method, in terms of correctness, robustness, and flexibility, and addresses issues (i) and (ii) satisfactorily.

Effects of Composition Type and Activator on Fly Ash-Based Alkali Activated Materials.

The compressive strengths of fly ash-based alkali-activated materials (AAM), produced using various activators of only sodium hydroxide, were measured. Fly ash-based AAM specimens, produced by mixing different kinds of fly ash and ground granulated blast-furnace slag (GGBFs) with an activator containing only sodium hydroxide, were cured at ambient temperature, and then placed in air for different numbers of days. The short- and long-term compressive strengths and shrinkage of fly ash-based AAM were measured and compared to one another. The effects of type of fly ash, alkali-equivalent content, GGBFs replace percentage, and ages on the compressive strengths and shrinkage of fly ash-based AAM were investigated. Even when different fly ash was used as the raw material for AAM, a similar compressive strength can be achieved by alkali-equivalent content, GGBFs replaces percentage. However, the performance of shrinkage due to different types of fly ash differed significantly.

Clincopathological characteristics and treatment outcomes of oral leukoplakia by carbon dioxide laser excision in the elderly patients.

BACKGROUND: Older age is one of the factors associated with malignant transformation of oral leukoplakia (OL). The purpose of this study is to analyze the clincopathological features and treatment outcomes of OL in the elderly patients. METHODS: The demographic data and histopathological results of the patients (age ≥ 65) who received carbon dioxide laser surgery for OL from 2002 to 2017 were analyzed statistically. RESULTS: There were 53 males and 16 females, with a mean age 71.2 ± 4.9. The follow-up time was 42.5 ± 35.2 months. In the univariate analysis, morphology, pathology, and area were found to be factors associated with postoperative recurrence. Among these factors, pathology and area were the independent predictive factors for recurrence in the multivariate logistic regression model. Malignant transformation occurred in 8 of 69 patients (11.6%). CONCLUSIONS: The pathological high-risk dysplasia and area of OL were the two prognostic factors for postoperative recurrence.

Treatment outcome of vocal cord leukoplakia by transoral laser microsurgery.

The aim of this study is to evaluate the treatment outcome and analyze the associated factors of postoperative recurrence in patients who received transoral laser microsurgery for vocal cord leukoplakia. The demographic, histopathological data were retrospectively reviewed and the factors associated with recurrence of vocal leukoplakia after surgery were analyzed statistically. A total of 44 patients, including 36 males and 8 females, with a mean age of 50.4 ± 13.4 years, were enrolled. All the patients received excision of the vocal leukoplakia by carbon dioxide laser (2-4 Watt, ultrapulse mode) under general anesthesia. No patients had malignant transformation after surgery. Postoperative recurrence occurred in 10 patients (22.7 %). Univariate analysis showed that patients who had the habit of cigarette smoking, alcohol drinking, and presence of gastroesophageal reflux disease tended to recur. Among these risk factors, presence of gastroesophageal reflux disease (odds ratio 8.43) was the independent prognostic factor for recurrence using multivariate logistic regression analysis. Carbon dioxide laser excision is effective for treating vocal leukoplakia that is still confined to dysplasia of any degree, with acceptable morbidity. This study suggests that the presence of gastroesophageal reflux disease is the prognostic indicator for postoperative recurrence of vocal leukoplakia. Aggressive treatment of reflux disease for those who have received surgical excision for vocal leukoplakia is indicated.

Stimulation of Proliferation and Migration of Mouse Macrophages by Type B CpG-ODNs Is F-Spondin and IL-1Ra Dependent.

Macrophage proliferation and migration are important for many facets of immune response. Here we showed that stimulation of macrophages with type B CpG oligodeoxynucleotides (CpG-B ODNs) such as CpG-ODN 1668 increased the production of anti-inflammatory cytokine interleukin 1 receptor antagonist (IL-1Ra) in a TLR9- and MyD88-dependent manner. The CpG-B ODNs-induced IL-1Ra increased macrophage migration and promoted macrophage proliferation by down-regulating the expression of a cell cycle negative regulator, p27 to increase cell population in the S phase. The induction of IL-1Ra by CpG-B ODNs was F-spondin dependent. Knockdown of F-spondin and IL-1Ra decreased CpG-B ODNs-induced macrophage migration whereas overexpression of IL-1Ra increased migration of those cells. These findings demonstrated novel roles for F-spondin and IL-1Ra in CpG-B ODNs-mediated cell proliferation and migration of macrophages.

Clinical characteristics of narrow-band imaging of oral erythroplakia and its correlation with pathology.

BACKGROUND: To analyze the clinical application of endoscope with narrow-band imaging (NBI) system in detecting high-grade dysplasia, carcinoma in situ, and carcinoma in oral erythroplakia. METHODS: The demographic, histopathological data, and NBI vasculature architectures of patients receiving surgical intervention for oral erythroplakia were retrospectively reviewed and analyzed statistically. RESULTS: A total of 72 patients, including 66 males and 6 females, with mean age of 54.6 ± 11.2 years, were enrolled. The odds ratio of detecting high-grade dysplasia, carcinoma in situ, and carcinoma by twisted elongated morphology and destructive pattern of intraepithelial microvasculature was 15.46 (confidence interval 95%: 3.81-72.84), and the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 80.95%, 78.43%, 60.71%, 90.91%, and 79.17%, respectively, which were significantly better than other two established NBI criteria (p < 0.001). CONCLUSIONS: Twisted, elongated, and destructive patterns of intraepithelial papillary capillary loop of NBI images are indicators for high-grade dysplasia, carcinoma in situ, and invasive carcinoma in oral erythroplakia.

Use of endoscopy with narrow-band imaging system in detecting squamous cell carcinoma in oral chronic non-healing ulcers.

OBJECTIVES: This study seeks to analyze the factors associated with the occurrence of squamous cell carcinoma in oral non-healing ulcers for more than 3 weeks and investigate the role of endoscopy with narrow-band imaging system (NBI) in detecting carcinoma in these lesions. METHODS: The demographic and clinicopathological data of patients receiving surgical interventions for chronic oral non-healing ulcers, and the intraepithelial papillary capillary loop patterns shown by NBI were retrospectively reviewed and analyzed. RESULTS: A total of 63 patients (41 males and 22 females) with mean age of 57.9 ± 16.7 years were enrolled. In univariate analysis, ulcers with induration, history of oral cancer, and intraepithelial microvasculature of NBI were factors associated with the occurrence of squamous cell carcinoma in oral non-healing ulcers. Multivariate analysis showed that the intraepithelial microvasculature of NBI was the only independent factor predicting the occurrence of carcinoma in oral chronic non-healing ulcers with sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 93.75 %, 91.49 %, 78.95 %, 97.73 %, and 92.06 %, respectively. CONCLUSIONS: Morphology of intraepithelial microvasculature of NBI, or twisted, elongated, and destructive pattern of intraepithelial papillary capillary loop, is the only independent factor associated with the occurrence of squamous cell carcinoma in oral chronic non-healing ulcers. Endoscope with NBI is a rapid, safe, and promising tool in detecting squamous cell carcinoma in oral chronic non-healing ulcers.

Synthesis and biological evaluation of helioxanthin analogues.

Helioxanthin and analogues have been demonstrated to suppress gene expression of human hepatitis B virus. In the continuous attempt to optimize antiviral activity, various structural motifs were grafted on the helioxanthin scaffold. Many such analogues were synthesized and evaluated for their anti-hepatitis B virus activity. Structure-activity relationships of these helioxanthin derivatives are also discussed. Among these new compounds, 15 exhibits the highest activity against HBV (EC50=0.06 µM). This compound can suppress viral surface antigen and DNA expression. Furthermore, viral RNA is also diminished while the core promoter is deactivated upon treatment by 15. A plausible working mechanism is postulated. Our results establish helioxanthin lignans as potent anti-HBV agents with unique mode of action. Since their antiviral mechanism is distinct from current nucleoside/nucleotide drugs, helioxanthin lignans constitute a potentially new class of anti-HBV agents for combination therapy.

Implications of morphologic patterns of intraepithelial microvasculature observed by narrow-band imaging system in cases of oral squamous cell carcinoma.

PURPOSE: To investigate the intraepithelial microvascular morphology of oral squamous cell carcinoma (OSCC) by using narrow-band imaging (NBI) and analyze whether the intraepithelial papillary capillary loop (IPCL) patterns correlate with infiltration depth and disease severity in OSCC. METHODS: The clinicopathologic data, morphology of vascular architecture as observed by NBI, and histopathology of patients with OSCC were retrospectively reviewed and analyzed. RESULTS: A total of 80 patients, including 73 males and 7 females with an average age of 54.18±12.23 years, were enrolled. Three patterns of intraepithelial microvasculature were revealed by NBI and differences in these three patterns were significant with regard to pathologic T-classification (p<0.0001), N-classification (p=0.00022), TNM stage (p<0.0001), lymphovascular invasion (p<0.0001), perineural invasion (p=0.000299), depth of tumor infiltration (p<0.0001), and tumor differentiation (p<0.0001). A cut-off point of tumor infiltration of 10.012 mm was best predicted for the destructive pattern of IPCL (sensitivity=100%, specificity=90.0%). CONCLUSIONS: Three different patterns of IPCL, showing step-wise increased severity according to pathologic parameters, were observed by NBI in cases of OSCC. The pattern indicating IPCL destruction with angiogenesis was associated with more advanced disease stage.

Recombinant viral protein VP1 suppresses HER-2 expression and migration/metastasis of breast cancer.

Breast cancer is one of the most common cancers in women worldwide and metastasis is the major cause of breast cancer death. Development of new therapeutic agents for inhibiting breast cancer metastasis is therefore an urgent need. We previously demonstrated that recombinant DNA-derived viral capsid protein VP1 (rVP1) of foot-and-mouth disease virus-induced apoptosis of MCF-7 breast cancer cells in vitro. Here, we investigated whether rVP1 exhibits any inhibitory effects on migration/metastasis and human epidermal growth factor receptor 2 (HER-2), a well-known biomarker for poor prognosis of breast cancer. The effects of rVP1 on cancer cell migration/invasion and metastasis were evaluated using Transwell migration assay and animal cancer models of metastasis. Western blotting, RT-PCR, flow cytometry, immunohistochemistry, and immunofluorescence staining techniques were used to investigate the effects of rVP1 on HER-2 and signal transduction mediators. Non-cytotoxic concentrations of rVP1-induced mesenchymal-epithelial transition and significantly suppressed AP-2α and HER-2 expression as well as the migration and invasion of a variety of breast cancer cell lines in a ß1-integrin-dependent manner in vitro. Gross and histopathologic examinations showed that rVP1 also suppressed metastasis of several breast cancer cell lines, including HER-2-overexpressing SK-BR-3 and BT-474 cells to lung, liver, or peripheral lymph node in orthotopic allograft/xenograft murine models. In addition, rVP1 significantly prolonged survival in breast cancer-bearing mice. Notably, no apparent side effects of rVP1 were detected, as shown by normal complete blood count levels and serum biochemistry profiles, including AST, ALT, BUN, and creatine. This study demonstrates that rVP1 suppresses the migration, invasion, and metastasis of breast cancer cells via binding to ß1 integrin receptor and down-regulation of AP-2α and HER-2 expression. The effectiveness of rVP1 on inhibiting migration/metastasis of breast cancer and HER-2 expression suggests that it may be suitable for serving as potential therapeutics for metastatic breast cancer particularly HER-2-overexpressing cancer.

Diagnostic significance of narrow-band imaging for detecting high-grade dysplasia, carcinoma in situ, and carcinoma in oral leukoplakia.

OBJECTIVES/HYPOTHESIS: To investigate the diagnostic accuracy of the established patterns of intraepithelial microvasculature of narrow-band imaging (NBI) in diagnosing upper aerodigestive tract neoplasm of the squamous epithelium for detecting high-grade dysplasia, carcinoma in situ, and carcinoma in oral leukoplakia. STUDY DESIGN: Retrospective case-control study. METHODS: Using histopathological findings as the standard, clinical diagnosis of oral leukoplakia and 3 different but established NBI criteria were compared and evaluated statistically. RESULTS: A total of 414 patients, including 365 males and 49 females, with mean age of 52.15 ± 10.75 years, were enrolled. The odds ratio of detecting high-grade dysplasia and carcinomatous lesions by twisted elongation of intraepithelial papillary capillary loop (IPCL) and IPCL pattern destruction was 95.53 (confidence interval 95%: 42.19-216.29), and the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 84.62%, 94.56%, 74.32%, 97.06%, and 93.0%, respectively, which were significantly better than the other two established NBI criteria (p < 0.001). CONCLUSIONS: The NBI images of twisted elongation of IPCL and IPCL pattern destruction are indicators of high-grade dysplasia or carcinomatous lesions in oral leukoplakia.

3'poly-G-tailed ODNs inhibit F-spondin to induce cell death and neurite retraction in rat embryonic neurons.

The effects and mechanism of action of oligodeoxyribonucleotides containing CpG motif (CpG-ODNs) on neuron cells are largely unexamined. Here, we found that CpG-A ODNs but not other types of CpG-ODNs induced neurite retraction and cell apoptosis of rat embryonic neurons in a TLR9-independent manner. These effects of CpG-A ODNs were primarily due to the poly-guanosine at the 3' terminus (3'G-ODNs). Pull-down analysis showed that 3'G-ODNs associated with transcription factor Y-BOX1 (YB-1) to facilitate the translocation of YB-1 into the nucleus via the nuclear localizing sequence of YB-1. YB-1 then interacted with the promoter of F-spondin directly at -45 and -1,375 sites as demonstrated by chromatin immunoprecipitation (ChIP) analysis. Binding of YB-1 to F-spondin promoter resulted in downregulation of F-spondin expression. Overexpression of F-spondin rescued the cell death and neurite retraction induced by 3'G-ODNs in embryonic neuron cells. Taken together, these findings suggest that 3'G-ODNs enhance nucleus YB-1 to inhibit F-spondin leading to cell death and neurite retraction of embryonic neuron cells.

Clinical appraisal of endoscopy with narrow-band imaging system in the evaluation and management of homogeneous oral leukoplakia.

PURPOSE: To evaluate the clinical usefulness of endoscopy with a narrow-band imaging (NBI) system for the evaluation and management of homogeneous oral leukoplakia. METHODS: The chart records, morphology of vascular architecture of NBI, and histopathology of patients with homogeneous leukoplakia were retrospectively reviewed and analyzed. RESULTS: A total of 160 patients, with an average age of 50.96 ± 10.25 years, were enrolled. In 35 cases of thin leukoplakia, only intraepithelium papillary capillary loop (IPCL) type I was shown by NBI, and only squamous hyperplasia was revealed pathologically. In 125 cases of thick leukoplakia, IPCL type I was found in 94, IPCL type II in 29, and IPCL type III in 2. The Kendall rank correlation between pathology and NBI images was significant (p < 0.0001). CONCLUSIONS: The dichotomous classification of homogeneous leukoplakia by NBI is meaningful, and endoscopy with the NBI system is a promising tool for the evaluation and management of homogeneous oral leukoplakia.

Use of endoscopy with narrow-band imaging system in evaluating oral leukoplakia.

BACKGROUND: The purpose of this study was to analyze the relationship between clinical features of oral leukoplakia using endoscopy with broadband white light, narrow-band imaging (NBI) illumination, and histopathology, and to discuss the clinical relevance of the NBI system. METHODS: Clinicopathologic data of patients receiving biopsy or excision after endoscopic examination for leukoplakia from April 2009 to December 2010 were reviewed and analyzed statistically. RESULTS: A total 154 patients, 130 men and 24 women (average age, 52.60 ± 10.86 years) were enrolled. The correlation between intraepithelium papillary capillary loop (IPCL) classification and stepwise increased severity of pathology (Pearson's r = .88, R(2) = 0.78) was significantly better than that of broadband white light images (p < .001). CONCLUSIONS: Flexible endoscopy can enhance detailed inspection of oral cavity mucosa and can be a powerful tool for examining oral leukoplakia. The IPCL patterns shown by NBI system can be helpful in detecting oral leukoplakia with higher grade dysplasia or invasive carcinoma.

Recombinant VP1, an Akt inhibitor, suppresses progression of hepatocellular carcinoma by inducing apoptosis and modulation of CCL2 production.

BACKGROUND: The application of viral elements in tumor therapy is one facet of cancer research. Recombinant capsid protein VP1 (rVP1) of foot-and-mouth disease virus has previously been demonstrated to induce apoptosis in cancer cell lines. Here, we aim to further investigate its apoptotic mechanism and possible anti-metastatic effect in murine models of hepatocellular carcinoma (HCC), one of the most common human cancers worldwide. METHODOLOGY/PRINCIPAL FINDINGS: Treatment with rVP1 inhibited cell proliferation in two murine HCC cell lines, BNL and Hepa1-6, with IC50 values in the range of 0.1-0.2 µM. rVP1 also induced apoptosis in these cells, which was mediated by Akt deactivation and dissociation of Ku70-Bax, and resulted in conformational changes and mitochondrial translocation of Bax, leading to the activation of caspases-9, -3 and -7. Treatment with 0.025 µM rVP1, which did not affect the viability of normal hepatocytes, suppressed cell migration and invasion via attenuating CCL2 production. The production of CCL2 was modulated by Akt-dependent NF-κB activation that was decreased after rVP1 treatment. The in vivo antitumor effects of rVP1 were assessed in both subcutaneous and orthotopic mouse models of HCC in immune-competent BALB/c mice. Intratumoral delivery of rVP1 inhibited subcutaneous tumor growth as a result of increased apoptosis. Intravenous administration of rVP1 in an orthotopic HCC model suppressed tumor growth, inhibited intra-hepatic metastasis, and prolonged survival. Furthermore, a decrease in the serum level of CCL2 was observed in rVP1-treated mice. CONCLUSIONS/SIGNIFICANCE: The data presented herein suggest that, via inhibiting Akt phosphorylation, rVP1 suppresses the growth, migration, and invasion of murine HCC cells by inducing apoptosis and attenuating CCL2 production both in vitro and in vivo. Recombinant protein VP1 thus has the potential to be developed as a new therapeutic agent for HCC.

Treatment outcome of dysplastic oral leukoplakia with carbon dioxide laser--emphasis on the factors affecting recurrence.

PURPOSE: To evaluate the associated factors of recurrence in patients who received laser surgery for dysplastic oral leukoplakia. MATERIALS AND METHODS: Clinicopathologic data of patients and their human papillomavirus genome status were used to analyze their association with recurrence after surgery. RESULTS: Of 114 enrolled patients, 90 were men and 24 were women with an average age of 49.7 ± 12.2 years, and follow-up was 1.75 to 9.1 years (mean, 3.4 ± 1.3 years). Recurrence after surgery occurred in 20 patients (17.5%). Thirteen patients had malignant transformation (11.4%). Twenty patients showed positivity for human papillomavirus (21.7%). Univariate analysis showed that patients who did not quit smoking or chewing betel quid after surgical treatment or whose oral leukoplakia took the form of widespread multiple-focus lesions, nonhomogeneous leukoplakia, and higher-grade dysplasia tended to have recurrence. Among these significant risk factors, continuous smoking after surgical treatment (odds ratio, 3.82) and widespread multiple-focus lesions (odds ratio, 4.54) were the independent prognostic factors for recurrence using multivariate logistic regression analysis. Those who did not quit chewing betel quid or smoking cigarettes were 19.8 or 9.7 times, respectively, more likely to develop recurrence than those who did quit. CONCLUSIONS: This study suggests that continuous smoking after surgical treatment and widespread multiple-focus lesions are the prognostic indicators for recurrence after laser surgery. Changes in oral habits can be of great importance to the outcome of laser surgery of dysplastic oral leukoplakia.

Postoperative recurrence as an associated factor of malignant transformation of oral dysplastic leukoplakia.

PURPOSE: To evaluate the clinically associated factors of malignant transformation in patients who received laser surgery for oral dysplastic leukoplakia. PROCEDURES: The clinicopathological data of patients receiving laser surgery for dysplastic leukoplakia from 2002 to 2008 and HPV genomic DNA extracted from paraffin-embedded specimens were analyzed statistically. RESULTS: A total 114 patients, 90 males and 24 females with an average age of 49.7 ± 12.2 years were enrolled. The follow-up period ranged from 1.07 to 7.43 years (mean 3.4 ± 1.3 years). Thirteen cases had malignant transformation (11.4%). Multiple-focus lesions, non-homogeneous leukoplakia, high-grade dysplasia and recurrence after laser treatment were significant factors in univariate analysis. Multivariate analysis showed that recurrence (relative risk = 9.40) was the independent prognostic factor for malignant transformation. CONCLUSIONS: Recurrence after laser treatment is an independent prognostic factor predicting malignant transformation of dysplastic oral leukoplakia.

High-dose bisacodyl plus water lavage compared with oral sodium phosphate as bowel preparation for outpatient colonoscopy.

BACKGROUND: The aim of this prospective study was to compare the efficacy and patient compliance of an oral high dose of bisacodyl plus water lavage and oral sodium phosphate in adults undergoing elective colonoscopy. METHODS: A total of 276 patients were randomized to receive an oral high-dose bisacodyl preparation (6 tablets of 5mg bisacodyl) plus water lavage (2 L of water) or an oral sodium phosphate (NaP) preparation (90 mL in divided doses). All endoscopic procedures were conducted in the afternoon. The endoscopist was blinded to the preparation the patient had received and graded the quality of colon cleansing as excellent, good, fair, poor, or failed. Patients' demographic data, indications for the procedure, and colonoscopic findings were recorded and compared. RESULTS: Significantly improved (p < 0.001) bowel preparation after NaP compared with bisacodyl plus water lavage was reported. The completion rate of colonoscopy was significantly lower (p < 0.001) in the bisacodyl group (70.6%) than in the NaP group (92.9%). The detection of polyps was significantly increased (p = 0.017) in the NaP group (43.1%) compared with the bisacodyl group (27.1%). A procedural indication of constipation was an independent predictor of failed preparation in the bisacodyl group (odds ratio, 4.8; 95% confidence interval, 1.9-12.0; p < 0.001). CONCLUSION: The quality of bowel preparation with oral NaP was better than that with bisacodyl plus water lavage.