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

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

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.

To establish a green fluorescent protein (GFP)-based cellular model for screening proteasome inhibitors from compounds including extracts from Traditional Chinese Medicinal herbs, we transfected A549 cells with lentivirus expression vector pGC-E1-ZU1-GFP, and selected clones stably expressing ZU1-GFP. The A549-ZU1-GFP cells were treated with PS-341 for 24 h, and the accumulation of GFP was analyzed by fluorescence microscope. We found that the fluorescence intensity of A549-ZU1-GFP cells treated with PS-341 was significantly increased as compared to the control. We screened for proteasome inhibitors from compounds including some from Traditional Chinese Medicinal herbs, and the data suggested a few compounds could be novel proteasome inhibitors.
Boronic Acids ; pharmacology ; Bortezomib ; Cell Line ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Models, Biological ; Protease Inhibitors ; isolation & purification ; pharmacology ; Pyrazines ; pharmacology

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 discuss the effects of pressure control (PC) and positive end-expiratory pressure (PEEP) incremental method lung recruitment maneuver (RM) on haemodynamics in piglets with acute lung injury (ALI) induced by paraquat (PQ) poisoning. Methods The ALI/acute respiratory distress syndrome (ARDS) model was reproduced by intraperitoneal injection of 20% PQ (20 mL) in 10 healthy female piglets, and they were randomly divided into PC lung RM group (RM1 group) and PEEP incremental method lung RM group (RM2 group), with 5 piglets in each group. Heart rate (HR), mean arterial pressure (MAP), and cardiac index (CI) were monitored by pulse-indicated continuous cardiac output (PiCCO) monitoring before model reproduction (baseline), on the time of successfully set up of model and at 5, 15 and 30 minutes after RM. At the same time the arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2) were recorded, and oxygenation index was calculated. Lung tissues were collected before model reproduction, on the time of successfully set up of model, and at 30 minutes after RM respectively, and pulmonary pathology changes were observed after hematoxylin and eosin (HE) staining under light microscopy. Results The HR, MAP, and PaCO2 on the time of successfully set up of model in both groups were increased obviously while CI, PaO2, and oxygenation index were decreased obviously as compared with those at baseline, all of which conformed to the expression of ALI/ARDS. With RM time extended, the HR in both groups was declined while MAP and CI were increased gradually. The HR and MAP at 5 minutes after RM of RM1 group were significantly lower than those of the RM2 group [HR (bpm): 126.8±5.2 vs. 134.0±3.8, MAP (mmHg, 1 mmHg = 0.133 kPa): 98.4±3.3 vs. 102.8±2.6, both P < 0.05]. The CI at 5 minutes and 15 minutes after RM of RM1 group was significantly higher than that of the RM2 group (mL·s-1·m-2: 56.7±5.0 vs. 46.7±6.7, 65.0±5.0 vs. 56.7±5.0, both P < 0.05). PaO2 and oxygenation index at 5 minutes after RM in both groups were significantly higher than those on the time of sucessfully set up of model. The PaO2 and oxygenation index were gradually decreased, and PaCO2 was increased with RM time extended, but no statistically significant differences at all time points were found between the two groups (all P > 0.05). The lung tissue in both groups showed a variety of pathological changes at 30 minutes after RM. The main performances were the loss of alveolar epithelial cells, the further wideness of alveolar interval and the distension of alveolar, and the part breakage of alveolar interval. The wideness of alveolar interval was more significant in RM2 group than that of RM1 group, and alveolar cleft was more common too. Conclusion Both PC and PEEP incremental method lung RM can improve the oxygenation of the piglets with ALI/ARDS induced by PQ, and the PC lung RM has less impact on haemodynamics.