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

AIM: To explore the effects of selenium-enriched Spirulina platensis (Se-SP) on proliferation of hepatocytes in rat hepatectomy. METHODS: Rat hepaectomy model was conducted using male Wistar rats. The rats were randomized into five groups: operation groups with 150 (H), 50 (M) and 15 (L) mg?kg-1?d-1 of Se-SP, placebo-control (P) and sham operation group (F). Activities of glutathione peroxidase (GPx) and thioredoxin reductases (TR) in hepatocytes were determined by chemical colorimetry. The expression index of proliferating cell nuclear antigen (PCNA) in hepatocytes was detected by immunohistochemistry, and the level of [3H]-TDR incorporation in regenerative hepatocytes was analyzed by radio-immunity. RESULTS: Activity of GPx and TR, PCNA expression index as well as [3H]-TDR insertion in hepatocytes (in vitro) were obviously higher (P

Objective To explore the effects of selenium enriched Spirulina platensis(Se-SP) on(antioxidation) and regeneration of rat hepatocytes after partial hepatectomy.Methods Sixty-seven percent(hepatectomy) was performed in rats after high dose(H) and low dose(L) Se-SP was given via gastric tube for 7 days.Then selenium(Se),glutathione peroxidase(GPx),superoxide dismutase(SOD) and(malondiaoldehyde)(MDA) in hepatocytes were determined.The results were compared with normal saline(giving) via gastric tube before hepatectomy(C) and sham operation(S) groups.Hepatocytes were obtained(before) operation and 24h after operation,and the expression index of proliferating cell nuclear antigen((PCNA)) in hepatocytes was detected by immunohistochemistry,and the DNA damage of hepatocytes induced by peroxide was analyzed by comet assay.Results After partial hepatectomy,Se content,GPx and SOD(activity),and PCNA expression index were obviously higher(P

AIM: To investigate the effect of Se-containing spirulina phycocyanin (Se-SPC) on liver injury of mice induced by carbon tetrachloride (CCl 4). METHODS: The mouse model was conducted by intragastric feeding with 2% CCl 4 oil for three times, meanwhile Se-SPC, spirulina phycocyanin (SPC) and Na 2SeO 3 were injected (ip) to various groups for 7 days. Then selenium (Se), glutathione peroxidase (GPx), superoxide dismutase (SOD), alanine aminotransferase (ALT), malondiaoldehyde (MDA) and nitric oxide (NO) levels in blood and liver were measured. RESULTS: The level of Se, GPx and SOD activities were obviously higher ( P

Objective To evaluate the characteristics and safety of capsule endoscopy for elderly patients.Methods The clinical data of 376 patients who underwent capsule endoscopy between May 2002 and September 2007 were retrospectively analyzed,and patients were divided into geriatric group(≥60 years old,n=134)and control group(

ObjectiveTo evaluate the effect of Rhubarb associated preparations (rhubarb or prescriptions of traditional Chinese medicine including rhubarb) on sepsis patients with acute gastrointestinal dysfunction (AGI).Methods The retrieval of databases from libraries including PubMed, Medline, Cochrane Central Register of Controlled Trials, CNKI, CBMdisc, Wan Fang Database, VIP database were searched to identify randomized controlled trials (RCTs) about Rhubarb associated preparations for treatment of sepsis patients with AGI from the foundation of the various databases to March 2016. And in the mean time, the references of the studies accepted were also retrieved. The retrieving and screening of literatures were performed independently by two researchers, the methodological quality and data extraction of the enrolled literatures were assessed by Jadad scale, and Cochrane Collaboration 5.3 software was used to perform Meta analyses to observe the effects of rhubarb associated preparations on gastrointestinal function score, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score and 28-day mortality in sepsis patients with AGI; the bias of published literatures was evaluated by funnel plot.Results ① Finally, 16 studies involving 1 171 patients (610 in rhubarb preparation group and 561 in the control group) were identified and enrolled. 12 studies had a Jadad score ≥ 3 and 4 studies < 3. The random method was used in classification of groups in all the studies in which the intergroup baseline data being comparable was clearly indicated. The blind method was applied in 5 contained RCTs.② The results of Meta-analyses showed that rhubarb associated preparation could improve gastrointestinal function score [mean difference (MD) = -0.52, 95% confidence interval (95%CI) = -0.55 to -0.48, P < 0.000 01], reduce the APACHEⅡ score (MD = -3.66, 95%CI = -5.00 to -2.33,P < 0.000 01) and 28-day mortality [odds ratio (OR) = 0.46, 95%CI = 0.30 to 0.71,P < 0.000 01] compared with those in the control group, the differences being statistically significant. No publication bias was seen in 16 literatures containing RCTs from the funnel plot.Conclusions Compared with the control group, the rhubarb associated preparations combined with conventional theraph can significantly improve the gastrointestinal function score, reduce APACHE Ⅱ score and 28-day mortality of sepsis patients with AGI, which suggests the rhubarb associated preparations have better efficacy. In addition, the result of sensitivity analysis has not substantially changed the results of Meta-analysis.

Objective To evaluate the diagnostic and therapeutic strategy of traumatic pulmonary pseudocyst (TPP).Methods Fifteen patients who were diagnosed and treated as TPP between January 2000 and November 2011 were studied retrospectively.Results Nonpenetrating chest trauma was the underlying cause in all cases.A typical sign shown on chest radiograph was a thin-walled cavitary lesion in 9 patients,6 patients accompanied by traumatic wet lung,with or without an air-fluid level.Serial radiological images of CT showed high resolution of the above lesions.Single TPP lesion occurred in 9 patients,and multiple TPP lesions in 6 patients.The size of the lesions was 5 -75 (32 ± 17) mm.The pseudocyst was located in the left lung in 5 patients(33％),located in the right lung in 7 patients (47％),located in bilateral lung in 3 patients (20％).All TPP patients were treated conservatively with no occurrence of complications.Conclusions TPP is an uncommon benign lesion secondary to thoracic trauma.CT scan is an optimal option for diagnosis and evaluation of TPP.Uncomplicated cases can take conservative treatment.For complicated patients,theraneutic strategy should be made individually.

Objective To evaluate the relationship between serum matrix metalloproteinase-9(MMP-9), white blood cell(WBC)count and type 2 diabetes mellitus with macroangiopathy and to investigate the mechanism of the protective effects of rusiglitazone(RSG)on blood vessel.Methods Serum MMP-9 was determined by ELISA in 30 normal controls and 80 type 2 diabetic patients(including 40 cases with macroangiopathy and 40 without maeroangiopathy).WBC count and other clinical parameters were also determined.32 type 2 diabetic patients received RSG(4rag qd)for 12 weeks.All parameters were determined after 4 weeks(17 patients)and after 12 weeks(all patients)to observe the changes in MMP-9,WBC and other parameters.Results In the diabetic patients,serum MMP-9 and WBC were markedly higher as compared with normal controls;and MMP-9 and WBC in patients with macroangiopathy[579(440-949)?g/L,(7.51?1.47)?10~9/L]were higher than those [324(275-423)?g/L,(6.22?0.79)?10~9/L]in the cases without macroangiopathy(P

Objective To explore the effect of nuclear factor-kappaB(NF-?B) on cardiac myocyte apoptosis and understand the molecular mechanism of decrease of heart function in septic shock rats.Methods Twenty Wistar rats were randomly divided into the septic shock group and the normal control group.The septic shock group(n=10) were fixed with anaesthesia and made into 1.5 cm slices just along the abdomen midline.Then the roots of appendices were returned to the belly cavity after being ligated and punctured 4 times with the No.14 pinhead.After that,the slices were sewn up layer by layer.After 12 hours,the rats were all sacrificed,the blood taken and the serum separated.In the end,the heart specimens of the septic shock group were set aside.Meanwhile,the normal control group were dealt with in the same way except that they were not subjected to cecal ligation and puncture.Then NF-?B protein levels in cardiac tissue and the index of cardiac myocyte apoptosis were measured.Results NF-?B protein levels in the septic shock group(9 cases were strong masculine gender,1 case was middle masculine gender)elevated significantly compared with the normal control group(8 cases were negative gender,2 cases were weak masculine gender) in cardiac tissue(P

Objective To investigate the role of white matter astrocytes and their specific protein S100A4 in sensory neurite outgrowth in vitro.Methods White matter astrocyte cultures expressing S100A4 were prepared. Dissociated adult dorsal root ganglion(DRG)cells were placed on the top of the astrocytes and co-cultured for 6,12, 18,24 hours.Small interfering S100A4 RNA was used to eliminate S100A4 expression.The growth of DRG cell neu- rites on S100A4-sileneed and S100A4-expressing astrocytes was compared.Results 12,18 and 24 hours after the co-culture with S100A4-expressing or S100A4-silenced astroeytes,neurite growth from the DRG cells was observed. Neurite outgrowth was significantly greater in S100A4 siRNA treated cultures compared to control siRNA treated white matter astrocyte cultures.Conclusion These findings suggest that white matter astroeytes are able to support axonal regeneration and,furthermore,that administration of small interfering S100A4 RNA provides strong additional support for axon regeneration.