Objective To evaluate the performance of enzyme linked immunosorbent assay (ELISA) kit in detection of Hepatitis B virus(HBV) markers by using improved and optimized method ,so as to provide a practical and feasible method and reagents for clinical laboratory .Methods ELISA test was used for the detection of HBV markers .The gradient dilution method was used to e‐valuate the lower limit .The verifiation of cut off value was carried out based on clinical and laboratory standards institute (CLSI) EP12‐A2 document .Samples with cut off values were collected to evaluate the precision ,including repeatability and intermediate precision .The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000SR chemiluminescence instrument .Results The lower detection limit of HBsAg ,HBsAb , HBeAg ,HBeAb and HBcAb were 0 .2 IU/mL ,20 mIU/mL ,1 NCU/mL ,0 .75 NCU/mL and 0 .05 NCU/mL respectively .The cut‐off value(C50 )± 20% concentration included the concentration range between C5 and C95 .The within‐run coefficient of variation (CV)≤15% ,in sandwich method the between‐run CV≤25% ,in competition method the between‐run CV≤35% .The positive and negative coincidence rates in accuracy and comparing with i 4000SR all were more than 95% and all κ>0 .75 .Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs .

Objective To clarifywhether HepG2 cells actively secrete Y-box binding protein-1 (YB-1) under stress conditions,and to investigate the pathological significance and mechanism of action of extracellular YB-1.Methods HepG2 cells were stimulated and treated by gradient concentrations of lipopolysaccharide (LPS) and adtiamycin,the supematant of the culture solution was collected by centrifugation,and the established chemiluminescence immunoassay (CLIA) was used for real-time quantitative determination of YB-1 level in the supernatant.The co-immunoprecipitation assay was used to detect whether extracellular YB-1 specifically bound to Notch3 receptor,and Western blot was used to measure the expression of Notch-NICD.The gradient concentrations of recombinant YB-1 were co-cultured with HepG2 cells,and MTT and migration assays were used to analyze the proliferation and invasion/metastasis of HepG2 cells.One-way analysis of variance was used for comparison of data between multiple groups.Results The results of CLIA confirmed that the level of extracellular YB-1 in the supematant was significantly higher than that in the control group (F =10.54,P ＜ 0.001),and the secretory expression of YB-1 reached its peak after 4 hours of stimulation (LPS:8 ng/ml;adriamycin:10 ng/ml).The results of co-immunoprecipitation assay and Western blot showed that extracellular YB-1 specifically bound to Notch3 receptor and upregulated the expression of the Notch3 receptor.MTT and migration assays showed that extracellular YB-1 significantly promoted the proliferation and invasion/metastasis of HepG2 cells (F =9.405,P ＜ 0.001).Conclusion Under the stress conditions induced by chemotherapeutics,HepG2 cells can actively secrete YB-1 via non-classical pathways.Extracellular YB-1 can specifically bind to Notch3 receptor and further up-regulate its expression,and then promote the proliferation and invasion/metastasis of HepG2 cells.This study lays a foundation for further clarifying the pathogenesis of hepatocellular carcinoma and investigating the biological relationship between extracellular YB-1 and malignant tumors.

OBJECTIVETo investigate GNE gene mutations in 5 Chinese patients with distal myopathy with rimmed vacuoles (DMRV).

METHODSFive patients with typical clinical and pathological features of DMRV were studied. All the 11 coding exons and the flanking intron sequences of GNE gene were amplified by PCR and sequenced. Four family members of case 5 were also examined for GNE gene mutations.

RESULTSAll the patients were identified to have different GNE gene mutations: Cases 1-4 had complex heterozygous mutations and case 5 had homozygous mutation. Six reported mutations had been identified, including 1 nonsense mutation (p.R8X) and 5 missense mutations (p.D176V, p.I298T, p.A591T, P.A631V, and p.V696M). A novel mutation (c.317T>C, p.I106T) was identified in case 2.

CONCLUSIONThis is the first report of p.R8X, p.I298T, p.A591T and p.V696M mutations in GNE gene in Chinese population, and a novel mutation p.I106T was identified. These findings further expand the clinical and genetic spectrum of DMRV in China.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA Mutational Analysis ; Distal Myopathies ; enzymology ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; Multienzyme Complexes ; genetics ; Mutation ; genetics ; Mutation, Missense ; genetics ; Young Adult

Objective To investigate the association between subclinical hypothyroidism and levels of systolic blood pressure (SBP), so as to provide evidence for the development of prevention strategy and understanding the etiology of hypertension. Methods The articles on the association of subclinical hypothyroidism and systolic blood pressure levels were retrieved by searching international and national databases from 1999 to 2010. The relationship between subclinical hypothyroidism and systolic blood pressure levels was assessed by meta analysis with Stata 11software. The weighted mean difference (WMD) and 95% confidence interval (CI) were calculated,and the publication bias was assessed by Begg's test and Egger's test. Results (1) There was significant difference in SBP levels between patients with subclinical hypothyroidism and normal subjects (WMD= 2.04 mm Hg, 95% CI: 0.64 to 3.45, P＜ 0.05 ). (2) Subgroup analysis indicated that there was significant difference seen in thyroid stimulating hormone (TSH) mean difference values ＜7 mU/L group(WMD=2.33 mm Hg,95%CI:0.60 to 4.06,P＜0.05) but not in the group that TSH mean difference values were ＞7 mU/L. There was significant difference seen in the Asian group (WMD=2.62 mm Hg, 95%CI: 1.69 to 3.55,P＜0.05) in the community group(WMD=2.77mm Hg, 95%CI: 1.61 to 3.93, P＜0.05) but not in the European group and or in the hospital group.There was significant difference in the cross-sectional group (WMD=2.77 mm Hg, 95%CI: 1.61 to 3.93, P＜0.05), but not in the case-control group. (3) Results from both Begg' s test and Egger's test did not show significant difference, indicating that there was no publication bias existed.Conclusion Subclinical hypothyroidism was associated with the elevated systolic blood pressure. In terms of the role of subclinical hypothyroidism that might serve as one of the potential risk factor for the elevated systolic blood pressure. Well designed and large sample-sized prospective studies were necessary to confirm the association between subclinical hypothyroidism and systolic blood pressure.Random controlled trials were also needed to study whether the treatment could lower the risk. Active treatment for subclinical hypothyroidism might be useful for prevention and treatment of hypertension.

Objective: To analyze the relationship between thrombin activatable fibrinolysis inhibitor ( TAFI ) and coronary heart disease ( CHD ), and to provide evidence for the prevention of CHD. Methods: The patients with CHD in Fushun Central Hospital in Liaoning Province were selected as the case group, the patients without CHD in the same hospital and period were selected as the control group. The demographic information and clinical examination results ( serum TAFI, lipid, glucose, etc. ) were collected to analyze the association between TAFI and CHD by logistic regression models.The multivariate logistic regression analysis was used to explore the relationship between TAFI and CHD. Results: There were 222 cases, including 100 cases of stable angina, 44 cases of unstable angina and 78 cases of acute myocardial infarction, and 222 controls. The median ages of cases and controls were 62 and 57 years old. The results of multivariate logistic regression analysis showed that serum TAFI>22.88 μg/mL ( P75 of controls ) was associated with the risk of CHD ( OR=1.619, 95%CI: 1.011-2.593 ), unstable angina ( OR=2.917, 95%CI: 1.433-5.939 ) and acute myocardial infarction ( OR=2.626, 95%CI: 1.007-6.847 ). Conclusion The high level of TAFI is related to CHD, unstable angina and acute myocardial infarction.

The study aims to explore the effects and mechanisms of water extract of Potentilla anserina (PA) on myelosuppression mice induced by cyclophosphamide based on metabonomics. The myelosuppressive mouse model was established by injected with cyclophosphamide and treated with water extract of PA. Thymus and spleen indexes, peripheral hemogram and bone marrow nucleated cells of each group was detected. Bone marrow pathology analysis was performed by hematoxylin-eosin staining. The levels of interleukin 3 (IL-3), interleukin 6 (IL-6), erythropoietin (EPO), granulocyte colony stimulating factor (GM-CSF), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in serum were measured. The changes of biomarkers and related metabolic pathways were analyzed by UPLC-Q-TOF/MS-based metabonomics. Animal experiments were approved by the Animal Ethics Committee of Southwest Minzu University. The high doses of PA could significantly improve the decrease of white blood cell (WBC), red blood cell (RBC) counts and hemoglobin (HGB) levels of mice induced by cyclophosphamide (P < 0.05), and significantly increase the number of nucleated cells and the area of hematopoietic tissue in femoral bone marrow. The medium and high doses of PA could significantly improve the serum levels of SOD, CAT, MDA, IL-6 and GM-CSF (P < 0.05), and have no significant effect on the expression of IL-3 and EPO (P > 0.05). Serum metabolomics analysis showed that the aqueous extracts of PA could alleviate myrosuppression by regulating the aminoacyl-tRNA, valine, leucine and isoleucine biosynthesis mediated by 13 different metabolites such as valine, leucine, asparagine and hydroxyisohexic acid. PA improve the inhibition of hematopoietic function in myelosuppression mouse, and its mechanisms may be related to anti-oxidation and promoting the expression of hematopoietic-related cytokines and regulating the related metabolic pathways.

OBJECTIVETo explore the linkage of the polymorphism at D17S1878 site with susceptible gene of essential hypertension.

METHODSForty-five pedigrees from the high prevalence region of the hypertension were collected. The polymorphism of D17S1878 site was genotyped with genetic analyzer and gene-scan software. Discordant sib pair analysis and affected sib pair analysis were used in linkage analysis.

RESULTSThere were significant differences in the age, male, alcohol-consuming, over-salt intake, average systolic pressure, average diastolic pressure, waist-to-hip-ratio, total cholesterol, high-density lipoprotein, and low density lipoprotein between the hypertensive sib and the normotensive sib (P < 0.05). There were eleven alleles at D17S1878 site, and the allele frequency was significantly different between the hypertensive and normotensive sibs (P < 0.05). Forty-three pedigrees were analyzed with affected sib pair analysis (t = 3.05, P < 0.05).

CONCLUSIONThe polymorphism of D17S1878 may be linked with susceptible genes of essential hypertension.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Genetic Linkage ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Phenotype ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Potassium Channels, Inwardly Rectifying ; genetics

BACKGROUNDMany studies have explored the diagnostic performance of soluble suppression of tumorigenicity-2 (sST2) for heart failure (HF), but the results are inconsistent. Here, we performed a meta-analysis to assess the role of sST2 in the diagnosis of HF.

METHODSWe searched PubMed, Web of Science, Cochrane Library, China National Knowledge Infrastructure, and Wanfang Database from inception to April 2015. Studies that investigated the diagnostic role of sST2 for HF were reviewed. The numbers of true-positive, false-positive, false-negative, and true-negative results were extracted to calculate pooled diagnostic odds ratio (DOR) with 95% confidence interval (CI) and the summary receiver operating characteristic curve and area under the curve (AUC). The Spearman correlation coefficient was used to check the threshold effect. The Cochran Q statistic (P < 0.05) and the inconsistency index (I2 > 50%) were used to assess the nonthreshold effect. Meta-regression was conducted to explore the source of heterogeneity; subgroup analysis showed the results in different subgroups. Finally, the Deeks' test was performed to assess the publication bias.

RESULTSNine articles including 10 studies were included in the meta-analysis. The pooled sensitivity was 0.84 (95% CI: 0.81-0.86), and pooled specificity was 0.74 (95% CI: 0.72-0.76). The summary DOR was 8.49 (95% CI: 4.54-15.86), and AUC was 0.81 (standard error: 0.03). The Spearman correlation coefficient identified the nonsignificant threshold effect (coefficient = 0.49, P = 0.148), but the nonthreshold effect heterogeneity was significant (Cochran Q = 58.52, P < 0.0001; I2 = 84.6%). Meta-regression found that characteristics of controls might be the suggestive source of nonthreshold effect heterogeneity (P = 0.095). Subgroup analysis found that DOR was 5.65 and 7.86, respectively for the controls of hospital patients and healthy populations. Deeks' test demonstrated that there was no publication bias (P = 0.616).

CONCLUSIONThe meta-analysis illustrated that sST2 might play a role in diagnosing HF.

Aged ; Female ; Heart Failure ; diagnosis ; Humans ; Interleukin-1 Receptor-Like 1 Protein ; physiology ; Male ; Middle Aged ; Publication Bias

OBJECTIVETo prepare antibodies (pAbs) against phosphorylated Y-box binding protein 1 (pYB-1), perform qualitative detection of the ascites/pYB-1 ratio in patients with hepatocellular carcinoma with pulmonary metastasis (HCC-PM), and assess the clinical significance of the ascites/pYB-1 ratio as a diagnostic biomarker for HCC-PM.

METHODSBioinformatic prediction and chemical synthesis was used to identify and generate the YB-1 polypeptide with phosphorylation at serine position 102 (KYLRSVGDG). Rabbits were immunized with the YB-1 polypeptide coupled to a carrier protein. Protein A affinity chromatography was used to prepare highly-purified pAbs.ELISA and SDS-PAGE were used to determine concentration and purity of the pAbs. A total of 109 ascites specimens were collected from patients (36 cases of HCC,44 cases HCC-PM, and 29 cases of liver cirrhosis) and concentrated to obtain the pYB-1. Western blotting was used to qualitatively detect pYB-1 in ascites. Regression analysis and receiver operating characteristic (ROC) curve analysis were used to assess the qualitative data.

RESULTSThe prepared pAbs had a concentration of more than or equal to 1:1 * 106 and high purity. The pAbs/YB-1S102 specifically recognized endogenous pYB-1S102. The pYB-1S102 detected in ascites specimens from patients with HCC and HCC-PM, and the positive rate of detection was 30.6% and 77.3% respectively (P < 0.01).The pYB-1S102 showed sensitivity of 77.3% and a accuracy rate of 73.8% for diagnosis of HCC-PM.

CONCLUSIONDetection of pYB-1S102 in ascites could be a useful biomarker for diagnosis and metastasis monitoring in patients with HCC.

Antibodies ; Biomarkers, Tumor ; Blotting, Western ; Carcinoma, Hepatocellular ; Humans ; Liver Cirrhosis ; Liver Neoplasms ; Neoplasm Metastasis ; Phosphorylation ; ROC Curve

OBJECTIVETo observe the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on biofilm of Staphylococcus epidermidis.

METHODThe serial dilution method was adopted to determine MIC of the combination of sodium houttuyfonate and erythromycin on S. epidermidis; the checkerboard method was used to evaluate the combination of sodium houttuyfonate and erythromycin on suspended bacteria of S. epidermidis; S. epidermidis biofilm was built in vitro, and XTT reduction assay was used to evaluate the effect of the combination of sub-MIC sodium houttuyfonate and erythromycin on the adhesion of S. epidermidis and bacterial metabolism inside the biofilm. Microscope was applied to observe the impact the single administration and combination of the two medicines under sub-MIC on biofilm morphology of S. epidermidis.

RESULTThe MIC of sodium houttuyfonate and erythromycin were 62.5, 7.812 5 mg x L(-1), respectively. The combination of 1/8MIC sodium houttuyfonate and 1/2MIC erythronmycin showed a synergistic effect on S. epidermidis. Sodium houttuyfonate, erythromycin and their combination had an inhibitory effect on the adhesion and metabolism of S. epidermidis biofilm bacteria, and made impact on the morphology of S. epidermidis biofilm.

CONCLUSIONThe sub-MIC sodium houttuyfonate and erythromycin have an inhibitory effect on S. epidermidis biofilm. The combination of sodium houttuyfonate and erythromycin shows a synergistic effect in inhibiting suspended bacteria and biofilm of S. epidermidis, particularly in inhibiting the metabolism of S. epidermidis biofilm bacteria and impacting the morphology of biofilm.

Alkanes ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; drug effects ; growth & development ; Dose-Response Relationship, Drug ; Drug Interactions ; Erythromycin ; pharmacology ; Microbial Sensitivity Tests ; Staphylococcus epidermidis ; drug effects ; physiology ; Sulfites ; pharmacology