This study aimed to construct full-length cDNA clones of the Japanese encephalitis virus (JEV). SA14-14-2 strain and discuss the feasibility of constructing chimeric viruses for exogenous gene expression based on the JEV genetic skeleton. Long-fragment RT-PCR techniques were applied to amplify JEV cD-NAs, and two amplified fragments with corresponding restriction endonuclease sites at both ends were cloned into the pACYC184 vector sequentially. Using standard molecular techniques, the enhanced green fluorescent protein (EGFP) gene was inserted into the 3' non-coding region of JEV as a reporter gene. After in vitro transcription and transfection procedures, wild-type JEV and chimeric JEV that expressed the EGFP as the reporter gene were successfully rescued. The recovered viruses were characterized by RT-PCR, plaque assays, and direct fluorescence microscopy. After six serial passage generations, the stability of the recovered viruses were studied in terms of virus growth characteristics and structural gene expression. The results showed that cDNA clones of rJEV and rJEV-EGFP were successfully constructed and rescued in BHK-21 cells after in vitro transcription and transfection. Each generation of the recovered viruses was stable and the chimeric virus rJEV-EGFP could stably express EGFP. The findings of this study indicate that both rJEV and rJEV-EGFP could be constructed and rescued in BHK-21 cells, and the JEV SA14-14-2 strain could be obtained as a viral vector to express foreign genes.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Encephalitis Virus, Japanese ; genetics ; metabolism ; Encephalitis, Japanese ; virology ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Vaccines, Attenuated ; genetics ; metabolism ; Viral Vaccines ; genetics ; metabolism

Acute Disease ; Antigens, CD ; analysis ; Biomarkers, Tumor ; analysis ; Chromosome Aberrations ; Diagnosis, Differential ; Humans ; Immunophenotyping ; Leukemia ; classification ; diagnosis ; genetics ; immunology ; Leukemia, Biphenotypic, Acute ; classification ; diagnosis ; genetics ; immunology ; Phenotype

Animals ; Brain ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Morphinans ; pharmacology ; Morphine Dependence ; metabolism ; Nitric Oxide Synthase Type I ; metabolism

Objective To investigate the clinical effect of anterior internal fixation plus vacuum sealing drainage in the treatment of ulnar and radial fractures of Gustilo type Ⅲ.Methods Twenty-eight patients with open ulnar and radial fracture of Gustilo type Ⅲ were managed from April 2007 to March 2014,and were divided into four groups(n =4).Group A were managed with external fixator and conventional changing dressings.Group B were managed with internal fixation and vacuum sealing drainage.Group C were managed with external fixator and vacuum sealing drainage.Group D were managed with internal fixation and conventional changing dressings.Result Twenty-eight cases were adopted telephone follow-up for 6 to 27 months.The soft tissue recovery time of each group respectively was (20.5 ± 2.37) days,(14.7 ±2.16) days,(15.6 ±2.17) days and(19.7 ±2.18) days.The hospital stay of each group respectively was (9.7 ± 2.54) weeks,(4.7 ± 1.46) weeks,(5.2 ± 2.34) weeks and 8.6 ± 2.16) weeks.The fracture healing time of each group respectively was (19.6 ± 2.74) weeks,(13.1±1.84) weeks,(18.1 ±2.54) weeks and (14.7 ± 1.74) weeks.There was significant difference of these data between the two groups(P ＜ 0.05).Conclusions Anterior internal fixation plus vacuum sealing drainage is a better way to treat ulnar and radial fractures of Gustilo type Ⅲ.The technique has short period,less complications,less painful and less expense.

Adult ; Aged ; Female ; Humans ; Infratentorial Neoplasms ; diagnosis ; surgery ; Male ; Meningioma ; diagnosis ; surgery ; Middle Aged ; Neurosurgical Procedures ; methods ; Supratentorial Neoplasms ; diagnosis ; surgery ; Treatment Outcome ; Young Adult

Objective To investigate the variation law of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values in patients with cerebral infarction, and to explore the relationship between the changes and the prognosis of cerebral infarct patients. Methods Sixteen patients with cerebral infarction were recruited and divided into 2 groups:good recovery and poor rehabilitation. ADC and FA values were calculated in infarct areas and control areas which were the regions with symmetrical position and the same area as infarct areas. The difference of ADC and FA values in patients at acute and earlier chronic stage between the two areas were analyzed. Results ①At acute stage, ADC values in infarct areas were lower than those in control areas (P<0.05). At early chronic stage, there was no significant difference of ADC values between infarct areas and control areas (P>0.05), moreover ADC values were higher than that at acute phase (P<0.05). ②FA values in infarct areas were lower than those in control areas at both acute and early chronic stage (P<0.05). At early chronic stage, FA values were lower than those at acute stage (P<0.05). ③There was no significant difference of ADC and FA values at both acute and early chronic stage between good recovery group compared with poor rehabilitation group (P>0.05). Conclusion There are certainly rules in changes of ADC and FA values in patients with cerebral infarction at acute and earlier chronic stage.

The historical surveillance results showed, there were 10 schistosomiasis cases in Huangshan City from 1994 to 2006. The survey in 2007 showed, the positive rates of blood examination for schistosomiasis in migrant workers and immigrant workers were 0.49% and 0.47% , respectively, but no schistosome-infected patients were detected by using the stool examination. An area with snails of 3 000 m~2 was found in the residence of the immigrant workers, but no infected snails were found. It is indicated that the mobile population has some impact on the transmission of schistosomiasis in the transmission-interrupted area. The surveillance and health education for the mobile population should be strengthened, and the imported infectious source should be prevented.

Objective To study correlation between systemic sclerosis(SSc)and plasma D-dimer and to reveal the probably rules of the fibrinolytic systems in SSc.Methods In 2013 January to 2014 January,cellected 32 patients with SSc and 35 with healthy controls to detected level of plasma D-dimer.Logistic and t student test were used for statistical analysis for correlation between SSc and pulmonary artery hypertension.Results When compared to healthy controls (0.28±0.04μg/ml),the level of plasma D-dimer were significantly increased in SSc patient (0.31±0.05μg/ml,t=1.997,P=0.008).After stratifying SSc patients according to disease subset,whereas patients with diffuse subset displayed substantially increased values (0.41±0.06μg/ml,t=2.051,P<0.001).The level of plasma D-dimer was associated with pulmonary artery hyper-tension (OR=4.38,95%CI=2.59~8.91,P=0.008).Conclusion Demonstrated that SSc patients with diffuse subset are characterized by increased plasma D-dimer values,reflecting a potential activation of fibrinolytic cascaded,which might finally predispose these patients to thrombotic complications and pulmonary artery hypertension.

Objective:To investigate the potential factors influencing the extent of coronary artery lesion in acute coronary syndrome (ACS) patients with an emphasis on the role of serum SIRT1.Methods:We assessed the clinical data from 81 ACS patients admitted to China-Japan Friendship Hospital. Serum SIRT1 was detected by enzyme linked immunosorbent assay (ELISA), and the extent of coronary artery lesion was evaluated by SYNTAX score before revascularization. All the patients were divided into two groups: high SYNTAX score (severe coronary artery lesion, n=38) and low SYNTAX score (moderate coronary artery lesion, n=43), by means of the median of SYNTAX score. Potential factors influencing SYNTAX score were analyzed through multiple linear regression analysis. Results:Compared with the low SYNTAX score group, patients in the high SYNTAX score group had higher serum SIRT1 level [379.38 (490.14) ng/L vs. 242.95 (173.85) ng/L, P<0.001] and frequency of coronary artery disease family history (42.11% vs. 20.93%, P=0.039). There was no statistical difference among other factors between the two groups. Serum SIRT1 was positively correlated with SYNTAX score in ACS patients ( R=0.452, P<0.010). Serum SIRT1 (ln adjusted), age and estimated glomerular filtration rate were independently correlated with SYNTAX score (ln adjusted) in multiple linear regression analysis (Adjusted R2=0.330, P<0.001). Conclusions:For the first time, we discussed the correlation of serum SIRT1 with extent of coronary artery lesion in ACS patients. Cardiologists should pay more attention to high-risk patients in order to improve the prognosis of ACS patients through timely revascularization strategies.

Four salts of ticagrelor, ticagrelor-3,5-dinitrobenzoic acid, ticagrelor-pyrazinamide, ticagrelor-D-proline and ticagrelor-L-proline were prepared by solvent suspension and liquid-assisted grinding to improve the solubility of ticagrelor. The compounds were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, elemental analysis, and the intermolecular salt-bonding forces were analyzed. The equilibrium solubility of salts and pure drug in hydrochloride buffer pH 1.2 and phosphate buffer pH 6.8 were measured by high-performance liquid chromatography. Ticagrelor was salted with 3,5-dinitrobenzoic acid, pyrazinamide, D-proline, L-proline all in a stoichiometric ratio of 1∶1; with the exception of ticagrelor-D-proline, the solubility of the other three salts provided significantly improved solubility in hydrochloride buffer pH 1.2, and the equilibrium solubility of ticagrelor-3,5-dinitrobenzoic acid was increased by approximately 1.7 folds as compared to pure drug. Salt-forming technology is convenient and can improve the solubility of ticagrelor.