Thermostable vacuum foam dried Newcastle disease vaccine: Process optimization and pilot-scale study.

Vacuum foam drying (VFD) has been shown to improve the thermostability and long-term shelf life of Newcastle Disease Virus (NDV). This study optimized the VFD process to improve the shelf life of NDV at laboratory-scale and then tested the optimized conditions at pilot-scale. The optimal NDV to T5 formulation ratio was determined to be 1:1 or 3:2. Using the 1:1 virus to formulation ratio, the optimal filling volumes were determined to be 13-17% of the vial capacity. The optimized VFD process conditions were determined to be at a shelf temperature of 25℃ with a minimum overall drying time of 44 h. The vaccine samples prepared using these optimized conditions at laboratory-scale exhibited virus titer losses of ≤ 1.0 log10 with residual moisture content (RMC) below 3%. Furthermore, these samples were transported for 97 days around China at ambient temperature without significant titer loss, thus demonstrating the thermostability of the NDV-VFD vaccine. Pilot-scale testing of the NDV-VFD vaccine at optimized conditions showed promising results for up-scaling the process as the RMC was below 3%. However, the virus titer loss was slightly above 1.0 log10 (approximately 1.1 log10). Therefore, the NDV-VFD process requires further optimization at pilot scale to obtain a titer loss of ≤ 1.0 log10. Results from this study provide important guidance for possible industrialization of NDV-VFD vaccine in the future. KEY POINTS: • The process optimization and scale-up test of thermostable NDV vaccine prepared through VFD is reported for the first time in this study. • The live attenuated NDV-VFD vaccine maintained thermostability for 97 days during long distance transportation in summer without cold chain conditions. • The optimized NDV-VFD vaccine preparations evaluated at pilot-scale maintained acceptable levels of infectivity after preservation at 37℃ for 90 days, which demonstrated the feasibility of the vaccine for industrialization.

Synergetic integration of catalase and Fe3O4 magnetic nanoparticles with metal organic framework for colorimetric detection of phenol.

Toxic phenol pollutants pose a great threat to the environment, it is urgent to develop an efficient and recyclable method to monitor phenol. Herein, we reported the synthesis of catalase-Fe3O4@ZIF-8 (CAT-Fe3O4@ZIF-8) through a novel amino-acid-boosted one-pot embedding strategy that synergically integrated catalase and magnetic Fe3O4 nanoparticles with ZIF-8. As expected, CAT-Fe3O4@ZIF-8 exhibited enhanced catalytic activity compared with Fe3O4@ZIF-8, CAT@ZIF-8 and catalase. Depending on the satisfactory performance of CAT-Fe3O4@ZIF-8, a colorimetric detection platform for phenol based on CAT-Fe3O4@ZIF-8 was constructed. The corresponding detection limit was as low as 0.7 µM, and a wide linear range of 5-100 µM was obtained. Besides, CAT-Fe3O4@ZIF-8-based colorimetric detection platform has been verified to possess high stability and recyclability. The proposed method was proven to have potential practical applications in the field of water treatment, which would advance efficient, recyclable monitoring of water quality.

☆DNA assembly technique simplifies the construction of infectious clone of fowl adenovirus.

Plasmid bearing adenovirus genome is generally constructed with the method of homologous recombination in E. coli BJ5183 strain. Here, we utilized Gibson gene assembly technique to generate infectious clone of fowl adenovirus 4 (FAdV-4). Primers flanked with partial inverted terminal repeat (ITR) sequence of FAdV-4 were synthesized to amplify a plasmid backbone containing kanamycin-resistant gene and pBR322 origin (KAN-ORI). DNA assembly was carried out by combining the KAN-ORI fragment, virus genomic DNA and DNA assembly master mix. E. coli competent cells were transformed with the assembled product, and plasmids (pKFAV4) were extracted and confirmed to contain viral genome by restriction analysis and sequencing. Virus was successfully rescued from linear pKFAV4-transfected chicken LMH cells. This approach was further verified in cloning of human adenovirus 5 genome. Our results indicated that DNA assembly technique simplified the construction of infectious clone of adenovirus, suggesting its possible application in virus traditional or reverse genetics.

Pentraxin 3, a Predicator for 28-Day Mortality in Patients With Septic Shock.

BACKGROUND: Useful biomarkers that can serve as prognostic predictors are of great value in clinical practice because of the complex individual response to sepsis. Pentraxin 3 (PTX3), as a multifunctional pattern-recognition molecule, has been reported to be closely associated with the severity of infectious diseases in intensive care units (ICU). The aim of this study was to investigate whether PTX3 could serve as a potential prognostic biomarker in patients with septic shock. MATERIALS AND METHODS: This single-center prospective observational study was conducted during May 2012-May 2015 in the ICU of Taizhou People׳s Hospital. We compared the clinical data and laboratory tests in surviving and deceased patients with septic shock within 28 days from admission. Potential independent prognostic factors for septic shock were analyzed by using univariate and multiple Cox proportional hazards regression analyses. RESULTS: A total of 112 patients admitted to the ICU with septic shock were enrolled in our study with an overall 28-day mortality of 25.9% (29 of 112 patients). PTX3 level was the only independent risk factor for the 28-day mortality by univariate and multivariate Cox analysis (hazard ratio = 3.87; 95% CI: 1.66-8.81, P = 0.004). The deceased patients had significant higher levels of PTX3 at the 4 different points (baseline, day 1, day 2 and day 3) versus the survivors (P < 0.001). Results from Kaplan-Meier curves and log-rank test revealed that high PTX3 level (above the median value) was statistically associated with a lower 28-day survival rate (P = 0.014). CONCLUSIONS: The baseline PTX3 level was an independent predictor for 28-day mortality in patients with septic shock.

Three Circulating LncRNA Predict Early Progress of Esophageal Squamous Cell Carcinoma.

BACKGROUND/AIMS: Previous studies revealed that circulating (either from plasma or serum) long non-coding RNA may predict the occurrence or prognosis of multiple human malignant tumors. In this study, we mainly explored whether circulating lncRNAs can be utilized as biomarkers predicting the development of human esophageal squamous cell carcinoma (ESCC). METHODS: LncRNA microarray was applied to screen the potential biomarkers for ESCC. Each group contained three individual plasma samples. A multi-stage validation and risk score formula detection were used for validation. RESULTS: Eleven dysregulated lncRNAs were obtained after Venny analysis. Further validation in a larger cohort including 205 ESCC patients, 82 patients suffering from esophagus dysplasia and 210 healthy controls confirmed that increased Linc00152, CFLAR-AS1 and POU3F3 might be potential biomarkers for predicting the early progress with an area under curve (AUC) of 0.698, 0.651 and 0.584, respectively. The merged AUC of the three factors and merged with CEA was 0.765 and 0.955, respectively. We also revealed that circulating levels of three lncRNAs were associated with poor post-surgery prognosis of ESCC patients. CONCLUSIONS: The three circulating lncRNAs might serve as potential biomarkers for predicting the early occurrence of ESCC.

Perioperative restricted fluid therapy preserves immunological function in patients with colorectal cancer.

AIM: To investigate the effect of perioperative restricted fluid therapy on circulating CD4(+)/CD8(+) T lymphocyte ratio, percentage of regulatory T cells (Treg) and postoperative complications in patients with colorectal cancer. METHODS: A total of 185 patients met the inclusion criteria and were included in the randomized clinical trial. These patients were divided into two groups according to receipt of either perioperative standard (S, n = 89) or restricted (R, n = 96) fluid therapy. Clinical data of these patients were collected in this prospective study. Perioperative complications and cellular immunity changes (CD4(+)/CD8(+) and Treg) were analyzed comparatively between the two groups. RESULTS: Both during surgery and on postoperative days, the total volumes of fluids administered in the R group were significantly lower than those in the S group (1620 ± 430 mL vs 3110 ± 840 mL; 2090 ± 360 mL vs 2750 ± 570 mL; 1750 ± 260 mL vs 2740 ± 490 mL; 1620 ± 310 mL vs 2520 ± 300 mL; P < 0.05). Decreased ratios of circulating CD4(+)/CD8(+) T lymphocytes (1.47 ± 0.28 vs 2.13 ± 0.26; 1.39 ± 0.32 vs 2.21 ± 0.24; P < 0.05) and Treg percentage values (2.79 ± 1.24 vs 4.26 ± 1.04; 2.46 ± 0.98 vs 4.30 ± 1.12; P < 0.05) were observed after surgery in both groups. However, in the R group, these values restored more quickly starting from postoperative day 2 (1.44 ± 0.24 vs 1.34 ± 0.27; 2.93 ± 1.08 vs 2.52 ± 0.96; P < 0.05). The proportion of patients with complications was significantly lower in the restricted group (36 of 89 vs 59 of 96, P < 0.01). CONCLUSION: Perioperative restricted intravenous fluid regimen leads to a low postoperative complication rate and better cellular immunity preservation in patients with colorectal cancer.