Based on the conserved region nucleotide sequences of five potato viruses/viroid(Alfalfa Mosaic Virus,ALMV;Cucumber mosaic virus,CMV;Cucumber mosaic virus-satellite,CMV-sat;Potato virus Y,PVY;Potato spindle tuber viroid,PSTVd) and one inner control(18S rRNA),the microarray containing specific oligonucleotide probes and PCR probes were designed and fabricated.The effects of probe concentration,hybridization time,hybridization temperature and spotting solutions on microarray hybridiazation were evaluated.Finally the specificity of optimized plant virus detection array was validated.No significant effect on hybridization signal intensity was observed when the concentration of the oligonucleotide probes ranged from 5 to 20 ?mol/L,there was a linear relationship between the concentration of PCR probe and hybridization signal intensity.The greatest signal intensity were obtained when hybridized at 45℃ for 4 h,and the oligonucleotide probes and PCR probes had a similar effect on microarray hybrization.Among the different spotting solutions,DMSO produced a good reproducibility.The plant virus could be detected specifically by oligonucleotide probe microarray and PCR probe microarray after optimization.

OBJECTIVETo introduce the framework of evidence-based practice with a case of castration-resistant prostate cancer (CRPC) as an example.

METHODSA clinical question was formulated according the clinical scenario. A systematic search was conducted for the published literature in the databases of PubMed, EMBASE, Cochrane Library, Clinical Trial Registries, and Web of Knowledge up to Dec 2014. The identified literature was reviewed for quality appraisal before the evidence was applied to clinical practice.

RESULTSThe treatment was effective and the patient achieved disease remission.

CONCLUSIONEvidence-based practice should be integrated with clinical scenario, current evidence, and patients' willingness, and follow a systematic framework.

Evidence-Based Medicine ; Humans ; Male ; Orchiectomy ; Prostatic Neoplasms, Castration-Resistant ; therapy

Objective To determine the effect of mitochondria on the protection of heat-shock proteins B1(HSPB1) from oxidative damage in rat cardiac cell.Methods HSPB1 gene-transfected rat cardiomyocytes cell line H9c2 (HSPB1 H9c2) and empty vector transfected H9c2 (control) were established,and treated by 0-1000?mol/L H_2O_2 for 2h.And then the cell morphology, mitochondrial membrane potential and endogenous reactive oxygen species (ROS) were detected. Results (1)HSPB1 inhibited the morphological changes induced by H_2O_2 markedly.(2)HSPB1 inhibited the loss of mitochondrial membrane potential induced by H_2O_2.Following the stimulation of 0,75,150,300,500,1000?mol/L H_2O_2,mitochondrial membrane potential in HSPB1 and control H9c2 cells were (10.0?0.11)vs (7.01?0.26),(9.11?0.17)vs (6.05?0.19),(7.69?0.28)vs (5.14?0.28),(6.95?0.13)vs (4.66?0.11),(6.61?0.20)vs (1.85?0.35),(6.60?0.05)vs (1.19?0.01),respectively (all P0.05).Conclusions HSPB1 protects rat cardiomyocytes cell line(H9c2) from oxidative damage,which suggests that stabilization of mitochondrial membrane potential and the decreased endogenous reactive oxygen species after oxidative stress may be involved in the protection of HSPB1 against oxidative stress in H9c2.

OBJECTIVETo study the epilepsy patients and their family members on their knowledge of the disease.

METHODSA 34-point questionnaire with 34 questions related to epilepsy knowledge was used for the survey on 170 pairs of epilepsy patients and their family members in Huashan hospital. Characters of the disease on the subjects were recorded.

RESULTSThe mean scores of the epilepsy knowledge of the patients and their family members were 16.5 +/- 8.2 and 16.1 +/- 8.5, respectively. The scores were quite low with no statistical difference between patients and their family members. The rate of correct answer in the urban subjects was obviously higher than those subjects living in the rural areas. All the subjects lacked the knowledge on the "cause of disease" when comparing with items as "diagnosis" and "treatment". Multivariate analysis showed that rural residents (P = 0.0001, OR = 52.963) and low education level (P = 0.0294, OR = 2.266) related to low epilepsy knowledge score among epilepsy patients. However, for family members, the factor related to low score was only living in the rural area (P = 0.0001, OR = 37.229).

CONCLUSIONEducation on the epilepsy knowledge should be strengthened, especially in the rural areas.

Adolescent ; Adult ; Epilepsy ; psychology ; Family Health ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Middle Aged ; Rural Health ; Surveys and Questionnaires

【Objective】 To establish a simple, economical and rapid method for the determination of methylene blue (MB) release in virus inactivation bag. 【Methods】 Based on the fluorescence energy transfer between MB and BSA-stabilized gold nanoclusters (BSA-AuNCs), the standard curve of MB determination was established by measuring the fluorescence quenching degree of MB to BSA-AuNCs in different concentrations to conduct the determination of MB release in virus inactivation bag. 【Results】 There was a good linear relationship between the MB concentration (cMB) and the fluorescence quenching degree of BSA-AuNCs[ (I0-I)/I0=0.018cMB+ 0.021(r=0.996)] when the fluorescence emission wavelength was about 620 nm and the cMB was in the range of (0.9-36) μmoL/L. The recovery of MB was 98.00% -101.95 % when applied to determine MB at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.73%, 0.81% and 0.77% respectively, and the obtained inter-day variation coefficients were 3.92%, 3.81%, and 4.73% respectively. There was no significant difference between the results measured by this method and those measured by combination of solid-phase extraction and spectrophotometry(P>0.05). 【Conclusion】 The fluorescence energy transfer method could achieve simple and rapid determination of MB release in virus inactivation bag with accurate and reliable results.

OBJECTIVETo investigate the clinical significance of neutralizing anti-interferon-alpha antibodies (NA) in chronic hepatitis B patients treated with recombinant interferon-alpha(rIFN-alpha).

METHODSOne hundred and eighty-one patients (128 male and 53 female) with histological proven chronic hepatitis B were treated with 5 MU recombinant interferon-alpha 1b (rIFN-alpha 1b) subcutaneously thrice weekly for 6 to 37 (median 10) months. For each patient, Specific detection of serum HBV DNA level with fluorescent-quantitative PCR, HBeAg with enzymoimmunoassay and NA with an antiviral neutralizing biological assay were performed during therapy.

RESULTSNA was found in 61 (33.7%) of 181 patients. At the end of treatment, complete-response was achieved in 17 (27.9%) of 61 patients with NA and in 54 (45.0%) of 120 patients without NA, respectively (chi2=4.979). For NA positive patients, the complete-response rate was significantly lower in those who had not achieved partial-response prior to or at the same time as NA occurred than in those who did (3.8%, 1/26, vs. 45.7%, 16/35, chi2 = 7.457). Moreover, it was lower in patients who either had 20pg/ml of serum HBV DNA or above or HBV DNA had being reduced by less than 60% recent 3 months, but higher in those who had less than 20pg/ml of HBV DNA and HBV DNA had being reduced by 60% or above (20.0%, 9/45, vs. 56.3%, 9/16, chi2 = 11.009).

CONCLUSIONNA may negate the antiviral effects of rIFN-alpha in chronic hepatitis B patients treated with rIFN-alpha, especially if they appear before partial-response or at the occasion at which serum HBV DNA level was not below 20pg/ml or HBV DNA had being reduced by less than 60% recent 3 months.

Antibodies ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; immunology ; therapeutic use ; Male ; Recombinant Proteins ; therapeutic use

AIMTo discuss the effect of Pitavastatin on angiogenesis in vivo and its mechanism in Klotho heterozygous deficient mice.

METHODSThe heterozygous deficient Klotho mice (kl +/-) and wild mice (kl +/+) from the same litter were used to establish the animal model of hind-limb ischemia and grouped into control and Pitavastatin group, respectively. Hind-limb blood flow was evaluated using Laser Doppler perfusion imager (LDPI) before treatment and after operation of hind-limbs. The capillaries in muscle of limbs were counted by means of CD-31 labeled immuno-fluorescence. The phosphorylation of Akt (Protein kinase B) in cells was measured by direct immunohistochemical technique. The expression of vascular endothelial growth factors (VEGFs) in muscle of limbs was assessed using Western blotting.

RESULTSAfter treatment of Pitavastatin, the blood flow in ischemic limbs of the Kl +/- and wild mice improved obviously, the ratio of blood flow area in ischemic limb to that in non-ischemic limb increased and the density of capillaries increased in ischemic limbs of the Kl +/- and wild mice. Pitavastatin enhanced the phosphorylation of Akt and the expression of VEGF in ischemic limbs of the Kl +/- and wild mice.

CONCLUSIONPitavastatin has the pro-angiogenesis effect in vivo and the VEGF-p-Akt-NO pathway may be involved in the mechanism of the effect of Pitavastatin.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Heterozygote ; Ischemia ; Male ; Mice ; Mice, Knockout ; Quinolines ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the survival of hand allograft under the state of immunosuppression and the pathological changes of rejection in the recovery process.

METHODSThe biopsies of the skin, nerve, muscle, tendon and bone tissue of hand allografts during different stages from 1 day to 7 months after operation were observed using routine histological technique.

RESULTSNo significant changes due to rejection in skin, nerve, muscle and bone tissue were observed. But different degrees of weak rejective changes were found on the wall of blood vessels; in the muscle and nerve the reactions were markedly stronger than those found in skin tissues.

CONCLUSIONSThe rejection in deep tissues should be monitored in controlling the rejection of hand allograft.

Adult ; Biopsy ; Graft Rejection ; pathology ; Hand Transplantation ; Humans ; Immunosuppression ; Male ; Skin ; immunology ; pathology ; Transplantation, Homologous

Objective To predict the value of advanced glycation end products (AGEs) in the risk and severity of coronary heart disease in diabetic patients.Methods Totally 120 cases were divided into 3 groups.Group A had no diabetes mellitus (DM) and no coronary atherosclerotic heart disease (CAD).Group B had DM without CAD.Group C had DM with CAD.The levels of AGEs,low density lipoprotein cholesterol (LDL-C),glycosylated hemoglobin,and the severity of coronary stenosis were detected.Receiver operating characteristic (ROC) curve was used to evaluate the sensitivities and specificities of AGEs for the diagnosis of DM with CAD.Results The highest level of AGEs,LDL-C,glycosylated hemoglobin and carotid plaque was in the C group,with a statistically significant difference (P ＜ 0.05).The severity of AGEs was significantly correlated with the severity of CAD and the Gensini score of AGEs (r =0.445).ROC curve showed that the sensitivity,specificity and area under ROC curve of AGEs determination of DM with CAD were 80.0％,75.0％,and 0.86,respectivity.Conclusions The level of AGEs is significantly correlated with atherosclerosis and prognosis in DM.The severity of CAD was higher in the patients with higher AGEs,and the incidence and severity of restenosis increased.

To explore the diagnostic value of copeptin (CPP) in cardiorenal syndrome (CRS) in rats and the association between CPP and impairment of heart and kidney, 60 male SD rats were randomly divided into blank control group (CK group), kidney failure group (SNX group), heart failure group (MI group), and CRS group. Heart and kidney function and their histology changes in rats from each group were detected. The correlation between serum CPP and heart and kidney function indexes was performed with Pearson correlation analysis. The HE staining of heart and kidney showed that the tissue lesion was more severe in CRS group than in SNX group and MI group. There was a significant positive correlation between serum CPP and brain natriuretic peptide (BNP) (r=0.638, P<0.05). No correlation was observed between serum CPP and cardiac function index (left ventricular systolic pressure, left ventricular diastolic pressure, left ventricular end-diastolic pressure) or renal function index (serum creatinine, urine creatinine, blood urea nitrogen) (r=0.512, 0.189, -0.063, 0.207, 0.290, 0.595, respectively, all P>0.05). The CPP level is associated with the degree of heart and kidney damage in CRS rats.