AIM:To investigate the regulatory effects and underlying molecule-mechanism of clonidine on learning and memory in rats with chronic cerebral ischemia.METHODS: Sprague-Dawley rats(n=45)were randomly divided into sham-operation group,cerebral ischemia model group and clonidine group,15 rats in each group.The chronic cerebral ischemia rat model was established by right middle cerebral artery occlusion for 2 h and reperfusion for 30 d. Clonidine was administrated by i.g.for 7 days in clonidine group.The ability of spatial reference memory of the rats with cerebral ischemia was tested by Morris water maze.The protein levels of extracellular signal-regulated kinase 1/2(ERK1/2),phosphorylated ERK1/2(p-ERK1/2), cAMP-response element binding protein(CREB)and phosphorylated CREB (p-CREB)were determined by immunohistochemistry and Western blot.RESULTS:The results of Morris water maze test showed that compared with the sham-operation group,the ability of spatial reference memory was obviously impaired in the cerebral ischemia model group.Compared with the cerebral ischemia model group,the ability of spatial reference memory in the clonidine group were improved.Compared with the sham-operation group, the protein levels of p-ERK1/2 and p-CREB in hippocampus were increased in model group(P<0.01).Compared with the cerebral ischemia model group,the protein levels of p-ERK1/2 and p-CREB in hippocampus were decreased in the clonidine group(P<0.01).CONCLU-SION:Clonidine improves the learning and memory abilities of the rats with cerebral ischemia, and ERK1/2 and CREB are involved in this process.

Objective· To investigate the association between neck circumference (NC) and cardiovascular risk factors among middle-aged and elderly people without diabetes in Jiading District in Shanghai.Methods· A cross-sectional study was conducted among 4 657 nondiabetic inhabitants aged 40 and above in Jiading District,Shanghai from August 2014 to July 2015.Clinical information collection,anthropometric measurements,and biochemical analyses were performed.The objects were divided into 4 groups according to the quartiles of NC in order to analyze association between NC and cardiovascular risk factors.Results· With increase of NC,the prevalences of abdominal obesity,insulin resistance,hypertension,and dyslipidemia all increased as well as waist circumference,body mass index (BMI),blood pressure,lipid profile,fast blood glucose,and HOMA-IR level (all Ptrend ＜0.01).Multiple Logistic regression analysis showed that individuals in Q2,Q3 and Q4 group had significantly higher risk of abdominal obesity,insulin resistance,hypertension,and dyslipidemia compared with those in Q1 group after age,sex,smoking,drinking,physical activity,BMI,waist circumference,systolic blood pressure,C-reactive protein,fast blood glucose,and lipid profile were corrected (all Ptrend ＜0.01).Conclusion· NC is positively and independently correlated with cardiovascular risk factors in middle-aged and elderly nondiabetic people in Jiading District in Shanghai.

Objective To investigate the bladder function in primipara and bipara within 1 week after delivery using urodynamic study.Methods Investigations on urodynamic changes were performed in 36 primipara volunteers and 12 bipara volunteers according to the recommendations of the International Continence Society(ICS).Fourteen women with upper urinary tract diseases but having normal lower urinary tract function,who had not experienced parturation were included as controls.Results Functional bladder volume(FBV)of primipara and bipara after delivery and normal desire cytometric capacity (NDCC)were respectively lower than those of control group(437?193)ml and(338?120)ml,however FBV and NDCC between primipara and bipara(310?154),(215?90)ml vs(243?141),(225?115) ml were not significantly different.The static Pure.max and Pure.clos.max of primipara and bipara were respectively higher than those of control group(87?7)cm H_2O(1 cm H_2O=0.098 kPa)and(78?8) cm H_2O(P

To study the effect of Yinghua Pinggan granule (YHPG) against influenza A/H1N1 virus in vivo and on the immunologic function of infected mice. The intranasal influenza virus infection was adopted in ICR mouse to establish the influenza virus pneumonia model. At the 3rd and 7th day after the infection, the lung index and pathologic changes in lung tissues of mice were detected. Realtime PCR and flow cytometry were employed to observe the virus load in lung tissues and the levels of CD4+, CD8+, and CD4+/CD8+ in peripheral blood. The result showed that at the 3rd and 7th day after the infection, YHPG (15, 30 g x kg(-1)) can significant decrease in the lung index and virus load in lung tissues of mice infected with influenza virus, alleviate the pathologic changes in lung tissues, significantly increase the levels of CD4+ and CD4+/CD8+ ratio and reduce the levels of CD8+ in whole blood. This indicated that YHPG can inhibit the influenza virus replication, alleviate pulmonary damage and adjust the weak immunologic function of infected mice, with a certain therapeutic effect on mice infected by H1N1 virus in vivo.
Animals ; Antiviral Agents ; administration & dosage ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; physiology ; Influenza, Human ; drug therapy ; pathology ; virology ; Lung ; pathology ; virology ; Male ; Mice ; Mice, Inbred ICR ; Virus Replication ; drug effects

OBJECTIVETo review the experience of staged total cavopulmonary connection (TCPC) in complex congenital heart diseases.

METHODSFrom June 1998 to March 2008, 22 patients underwent staged TCPC for complex congenital heart diseases. Among them, 9 were univentricular and pulmonary artery valve stenosis; 3 were univentricular and pulmonary artery atresia; 1 was transposition of great arteries, crisscross heart and pulmonary artery valve stenosis; 1 was complete atrioventricular canal defects, left ventricular hypoplasia, pulmonary artery atresia and atrioventricular valvular regurgitation; 1 was complete atrioventricular canal defects, left ventricular hypoplasia, pulmonary artery valve stenosis and atrioventricular valvular regurgitation after Glenn procedure; 1 was mirror image dextrocardia, single ventricle, pulmonary artery atresia, major aortopulmonary collateral arteries (MAPCAs) and right pulmonary arteriovenous fistula after Glenn procedure; 4 were tricuspid atresia and pulmonary artery valve stenosis; 1 was tricuspid atresia and pulmonary atresia; 1 was mirror image dextrocardia, double-outlet of right ventricle, left ventricular hypoplasia, pulmonary artery valve stenosis, tricuspid incompetence, and MAPCAs. Among them, 5 patients received systemic-to-pulmonary artery shunt, bidirectional Glenn procedure and TCPC. Seventeen patients received bidirectional Glenn procedure, the mean age was (5.9+/-4.4) years old. Pulmonary artery pressure pre-Glenn procedure was 17 to 20 mm Hg (1 mm Hg=0.133 kPa). Atrioventricular valve incompetence in 3 patients. Nakata index was less than 200 mm2/m2 in 4 patients before the first stage operation. The age of TCPC procedure was (9.6+/-4.9) years old, the interval time was (3.7+/-1.2) years.

RESULTSThere was one in-hospital death, the mortality was 4.5%. The patient with univentricular and pulmonary atresia, received systemic-to-pulmonary artery shunt, bidirectional Glenn procedure and TCPC and died of pneumorrhagia. Other patients were recovered well, postoperative central venous pressure was 12 to 18 mm Hg, percutaneous oxygen saturation was 90% to 96%. The cardiac function were in NYHA class I to II.

CONCLUSIONSThe staged TCPC was a good procedure in high-risk Fontan candidates. The results were satisfactory for those patients. This staged strategy may extend the operative indications for the Fontan procedure.

Adolescent ; Anastomosis, Surgical ; methods ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Heart Bypass, Right ; methods ; Heart Defects, Congenital ; surgery ; Humans ; Male ; Pulmonary Artery ; surgery ; Retrospective Studies ; Treatment Outcome ; Venae Cavae ; surgery ; Young Adult

Adult ; Catheter Ablation ; methods ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Low Back Pain ; surgery ; Male ; Middle Aged

OBJECTIVETo evaluate the initial clinical characteristics, the response to treatment, and the outcome in adult patients with Evans syndrome.

METHODSThe clinical data of 84 adult patients (20 males, 64 females) with Evans syndrome diagnosed at our center between 1984 and 2007 were retrospectively analyzed.

RESULTSThe patients were followed up for a median duration of 17.5 (0.03 - 140) months. All the patients initially received intravenous steroids with or without intravenous immunoglobulin (IVIG). Forty-seven patients were treated with corticosteroids alone initially. Complete remission (CR) and partial remission (PR) were achieved in 38 of the patients, but 92.1% of them relapsed during a median follow-up of 12 months. Twenty-eight patients who were resistant to corticosteroids therapy or with severe bleeding were subsequently administered immunosupressive agents. CR and PR were obtained in 89.3% of them. Within a median follow-up of 8 months, 84% of these patients relapsed.

CONCLUSIONSEvans syndrome is a chronic and easy to recurrent disease, which is often refractory to conventional therapy. Treatment with combination agents might be a useful therapeutic approach to the patients.

Adrenal Cortex Hormones ; Adult ; Anemia, Hemolytic, Autoimmune ; Follow-Up Studies ; Humans ; Remission Induction ; Retrospective Studies ; Treatment Outcome

To construct a hepatitis C virus (HCV) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, Huh7.5 and Huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of HCV replication and selection studies for putative anti-HCV drugs. Site-directed mutagenesis was used to delete the Core-E1-E2-p7-NS2 fragment (about 3090 bp) from plasmid pJ6JFH1BlaRL. The resultant trianglepJ6JFH1BlaRL plasmid was digested with AgeI and AvrII to release the cDNA fragment (hereafter, referred to as fragment L) containing partial 5'-untranslated region (UTR), the first 12 amino acid (aa) of HCV Core coding sequence, full-length coding sequences for the blasticidin-resistance gene, Renilla luciferase, foot-and-mouth disease virus (FMDV) 2a antiprotease and ubiquitin, and partial coding sequence for HCV NS3. To generate the monocistronic replicon, pSGRmJFH1BlaRL, fragment L was ligated into the pSGR-JFH1 vector that had been digested with AgeI and AvrII to remove the partial 5'-UTR, the first 19 aa of HCV Core coding sequence, the full-length coding sequence for the neomycin phosphotransferase II gene, the internal ribosomal entry site from encephalomyocarditis virus, and partial HCV NS3 coding sequence. A replication-defective mutant replicon, pSGRmJFH1BlaRL/GND, was constructed by a similar procedure using the pSGR-JFH1/GND vector. Fragment L was confirmed in both constructs by sequencing. Replicon RNAs were prepared from XbaI-linearized plasmid DNA templates with Invitrogen's T7 MEGAscript kit, and were purified by DNase I treatment and LiCl precipitation. RNAs were quanti?ed by optical density, and the quality and concentration were con?rmed by agarose gel electrophoresis. Replicon RNAs were transfected into Huh7.5 and Huh7.1 cells using Invitrogen's DMRIE-C transfection reagent at a ratio of 5 mug of lipid to 1mug of RNA. Time course assay of Renilla luciferase activity indicated the replicon's replication function. The pSGRmJFH1BlaRL monocistronic replicon and pSGRmJFH1BlaRL/GND replication-defective mutant replicon were successfully constructed. The pSGRmJFH1BlaRL replicon was replication-proficient in Huh7.5 and Huh7.1 cells, with replication peaking at 72 hours post-transfection and decreasing after 96 hours. No replication was detected at any time point post-transfection for the defective mutant replicon. A monocistronic replicon of HCV genotype 2a was constructed and shown to be replication-proficient in human hepatocarcinoma cell lines.
Cell Line, Tumor ; Genetic Vectors ; Genome, Viral ; Genotype ; Hepacivirus ; genetics ; Humans ; Mutagenesis, Site-Directed ; RNA, Viral ; Transfection ; Virus Replication

OBJECTIVETo investigate the impact of interferon-stimulated exonuclease 20 kDa (ISG20) on replication of genotype 2a hepatitis C virus (HCV) subgenomic replicon RNA and infectivity of the cell culture-derived HCV strain JFH1 to determine the potential of exogenously expressed ISG20 as an anti-viral therapy of chronic hepatitis C.

METHODSPlasma vectors containing wild-type (WT) ISG20 or a catalytically-inactive mutant ISG20m were transiently transfected into Huh7, Huh7.5 and HEK293 cells, and the replication of a monocistronic subgenomic JFH1 RNA replicon, SGRm-JFH1BlaRL, was measured. Huh7.5 cells stably expressing ISG20, ISG20m, or the control vector were established by transducing replication incompetent pCX4-Bsr-myc retroviruses encoding WT ISG20, D94G mutant ISG20, or the empty vector, respectively, and selecting with 5 mug/mL of blasticidin for approximately three weeks. The stable Huh7.5 cells were then transfected with HCV replicon RNA and infected with cell culture-derived HCV to investigate inhibition capacity of ISG20 against HCV.

RESULTSHuh7.5-ISG20, Huh7.5-ISG20m, and Huh7.5-Bsr controls cells stably expressing ISG20, ISG20m, or the control vector, respectively, were constructed successfully; the ectopically expressed ISG20 and ISG20m were distributed in both nucleus and cytoplasm, as detected by immuno uorescence. SGRm-JFH1BlaRL replicated efficiently and with similar kinetics in the Huh7.5-Bsr and Huh7.5-ISG20m cells, with expression levels plateauing at 48-96 h post-transfection. In contrast, at all time points examined, SGRm-JFH1BlaRL replication was 9.1% to 16.7% in the Huh7.5-ISG20 cells. The Huh7, Huh7.5 and HEK293 cells transiently expressing ISG20 also showed 16.7% to 25.0% of HCV replication that the respective controls. In addition, the amount of infectious progeny JFH1 virus released in culture supernatants was 9.1% to 12.5% from the Huh7.5-ISG20 cells than from the Huh7.5-Bsr and Huh7.5-ISG20m cells at 48-72 h post-infection, and the latter two cultures produced similar JFH1 virus yields. Finally, the expression of HCV core protein was also lower in the Huh7.5-ISG20 cells, as detected by immunoblot analysis.

CONCLUSIONExogenous expression of ISG20, either in a transient or stable manner, suppresses not only replication of genotype 2a HCV RNA replicons but also JFH1 virus propagation in cultured hepatocytes. The exonuclease activity of ISG20 is required for its antiviral activities against HCV.

Antiviral Agents ; pharmacology ; Cell Line ; Genome, Viral ; HEK293 Cells ; Hepacivirus ; genetics ; Humans ; RNA, Viral ; genetics ; Replicon ; Virus Replication ; drug effects