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【Objective】 To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. 【Methods】 Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. 【Results】 The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. 【Conclusion】 This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.
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The healthy development of science and technology (Sci. & Tech) innovation is inseparable from Sci. & Tech ethics. The innovation subject is the driving force of Sci. & Tech innovation. The improvement of Sci. & Tech ethical management ability is conducive to improving the China’s Sci. & Tech ethics governance system. Therefore, this paper deeply analyzed the current status of Sci. & Tech ethics management of the 76 innovation subjects from five aspects, and found that the construction of Sci. & Tech ethics management of Chinese innovation subjects has achieved initial results, but there was still room for further improvement. Based on the problems existing in the Sci. & Tech ethics management of innovation subjects, this paper proposed six strategic suggestions that strengthening the construction and research of artificial intelligence ethics management systems, establishing and improving ethics committees and management systems, actively implementing the registration of Sci. & Tech ethics management information, regularly organizing and participating in various forms of Sci. & Tech ethics training, attaching importance to the reserve and cultivation of composite Sci. & Tech ethics management talents, and cooperating to establish a Sci. & Tech ethics case library, with a view to enhancing the Sci. & Tech ethics management ability of Chinese innovation subjects to adapt to the needs of the healthy development of Sci. & Tech in China.
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This paper discusses the application of MRN combined with 3D printing in SNM in neurogenic bladder patients with special comorbidities. In this study, 21 cases of neurogenic bladder patients with special comorbidities involved, including 14 cases into the control group and 7 cases in the experimental group. Traditional X-ray guidance was used in the control group, while the MRN combined with 3D printing navigation template positioning was used in the experimental group. Compared with those in the control group, the X-ray fluoroscopy, number of punctures, puncture time, intraoperative adjustment and testing time were significantly decreased in the experimental group. The difference of implantable pulse generator implantation rates between the two groups was statistically significant.
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DNA damage repair (DDR) genes are overexpressed in liver cancer. Studies have found that DDR is closely related to the occurrence and development of liver cancer. There is a synergistic effect between DDR inhibitors and chemotherapy and targeted drugs for liver cancer. DDR inhibitors can enhance the sensitivity of radiotherapy. Some DDR genes can be used as biomarkers for the prognosis of liver cancer.
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Objective:To investigate the prognostic value and related factors of heart-type fatty acid binding protein (H-FABP) in patients with heart failure.Methods:A total of 877 consecutive patients who were admitted to heart failure care unit of Fuwai hospital and diagnosed as heart failure from July 2015 to July 2017 were enrolled in this study. Baseline serum H-FABP concentration was measured by fluorescence lateral flow immunoassay. According to serum H-FABP levels, patients were divided into three groups: low H-FABP group (H-FABP≤4.04 ng/ml, n=292), middle H-FABP group (H-FABP 4.04-7.02 ng/ml, n=292) and high H-FABP group (H-FABP≥7.02 ng/ml, n=293). The general clinical characteristics were collected and compared among the three groups. According to whether heart failure was caused by coronary artery disease or not, patients with heart failure were divided into ischemic heart failure and non-ischemic heart failure. Multivariate linear regression analysis was performed to explore the independent risk factors of H-FABP. The primary endpoint events were the composite of all-cause death or heart transplantation. Multivariate Cox regression analyses, receiver operating characteristic (ROC) curves, risk prediction tests with multivariate Cox regression model and Kaplan-Meier analyses were conducted to investigate the relationship between H-FABP and the prognosis of heart failure. Results:Multivariate linear regression analysis showed that age, coronary artery disease, alanine aminotransferase, uric acid and N-terminal pro-B type natriuretic peptide (NT-proBNP) were positively associated with H-FABP (β=0.012, 0.238, 0.001, 0.345 and 0.063 respectively,all P<0.05), while female, hemoglobin, albumin, sodium, and estimated glomerular filtration rate (eGFR) were negatively associated with H-FABP (β=-0.184, -0.006, -0.016, -0.034 and -0.006 respectively, all P<0.05). One hundred and nineteen patients (13.6%) lost to follow-up, and 246 patients (32.5%) suffered from all-cause death or heart transplantation during the median follow-up duration of 931 (412-1 185) days. Multivariate Cox regression analysis showed that baseline H-FABP (log 2H-FABP) level was the independent predictor of all-cause death or heart transplantation in patients with heart failure ( HR=1.39, P<0.001). ROC curves showed that baseline H-FABP was a predictor of all-cause death or heart transplantation in patients with heart failure within 3 months, 1 year and 2 years (areas under the curves were 0.69, 0.69 and 0.71 respectively), and the best cut-off values were 5.85 ng/ml, 6.54 ng/ml and 6.54 ng/ml respectively. Risk prediction test with multivariate Cox regression model showed that baseline H-FABP could provide additional prognostic value in predicting all-cause death or heart transplantation for patients with heart failure on top of basic model and baseline NT-proBNP ( P<0.001). Taking 6.54 ng/ml and trisected levels of H-FABP as cut-off values respectively, Kaplan-Meier analyses showed that the survival rates were significantly different among the two or three groups ( P<0.001). Subgroup analyses showed that baseline H-FABP (log 2H-FABP) level was an independent predictor of all-cause death or heart transplantation in patients with ischemic heart failure ( HR=1.74, P<0.001), as well as in patients with non-ischemic heart failure ( HR=1.28, P=0.027). Conclusions:Age, sex, coronary artery disease, hemoglobin, albumin, alanine aminotransferase, sodium, eGFR, uric acid and NT-proBNP are associated with H-FABP level. Baseline H-FABP level is an independent predictor of all-cause death or heart transplantation in patients with heart failure. On top of basic model and baseline NT-proBNP, baseline H-FABP could provide additional prognostic value in predicting adverse events for patients with heart failure.
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Objective:To study the experience of frontline medical workers in COVID-19 quarantine points.Methods:Purposive sampling method was employed and 12 frontline medical workers in COVID-19 points of Zhaoyuan City were selected for semi-structured and in-depth interview. Colaizzi′s seven-step analysis method was adopted to analyze and categorize the data.Results:The real experience of frontline medical workers in quarantine points were classified into five themes: professional mission and pride, inadequate initial response in dealing with public health emergencies (shortage of personal protective equipment,inadequate preparation of medical workers and imperfect settings of quarantine points), having some negative emotions (worrying about being infected and feeling fearful for family members), facing complex and diverse working difficulties (repugnance and diversified demands of people quarantined, heavy workload, discomfort from personal protective equipment and occupational exposure), and gaining support and appreciation (getting support from epidemic control and prevention departments as well as medical systems and winning appreciation from people quarantined).Conclusions:The frontline medical workers are lack of experience in dealing with public health emergencies and have some psychological and working pressure. Administrative department should further complete the emergency plan of centralized isolation medical observation, strengthen the contingency management of public health emergencies, attach importance to the mental health of medical workers, optimize management methods and establish flexible work schedules.
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Objective Toinvestigatetheapplicationvalueofthreedimensionalliveraccelerationvolumeacquisition(3D LAVA) multiphasedynamiccontrastenhancedMRIinBudd-Chiarisyndrome(BCS).Methods Aretrospectiveanalysiswasperformedforthe clinicaldataof49patientswithBCSwhounderwent3DLAVA multiphasedynamiccontrastenhanced MRIandwereconfirmedby DSA.Theappearanceofinferiorvenacava,hepaticvein,accessoryhepaticvein(AHV)andcollateralcirculationonbothDSAand3D LAVA wereobserved.BCStypeandAHVnumberswereanalyzedusingtheχ2test.3DLAVAandDSAinthemeasurementofthe mean diameter of A H V were compared by Student’s t test.The Kappa statistics was calculated to quantify the consistency between thetwomethodsinthedetectionofAHV.Results Amongthe49patients,therewasnosignificantdifferencebetweenMRIandDSA inBCSclassification (P=0.694).MRIandDSAshowedstatisticalsignificances(P<0.01)inthenumberofAHVandthemeancaliber of the A H V respectively.T he Kappa coefficient (K=0.621 ) de m onstrated good consistency for the tw o m ethods in detecting the numberofAHV.3DLAVAshowedmoreadvantagesincollateralcirculationthanDSA.Conclusion 3DLAVA mayaccuratelydetect hepaticvein,inferiorvenacava,AHV,andextrahepaticcollateralcirculationinBCSandcanhelptoevaluatethepatient’sconditionas wellastoselectthealternativemodalityininterventionaltherapy.
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Objective ToexplorethevalueofMRIindifferentiatingbetweenmass-formingchronicpancreatitis(MFCP)andpancreatic carcinoma(PC).Methods MRIdataof19caseswith MFCP,36caseswithPCand30normalcontrolcasesconfirmedbypathology orfollow-upwereanalyzedretrospectively.AllofthesubjectsunderwentroutineMRIandDWIscan.MRIcharacteristicsofdiseases andnormalpancreaswereanalyzed,andADCvalueswerecomparedamongthethreegroups.Results TheaverageADCvalueofthe MFCPgroupwas(1.41±0.25)×10-3 mm2/s,higherthanthatofthePCgroup (1.13±0.11)×10-3 mm2/s(P<0.05),andlower thanthatofthenormalcontrolgroup(1.50±0.27)×10-3 mm2/s(P<0.05).IntheT2WI,enhancedscanningarterialphase,andenhanced scanningportalphase,thesignalcharacteristicsofthelesionswerestatisticallydifferentbetweentheMFCPandPCgroup (P<0.05).The sensitivityandspecificityofthecombinationT2WI,enhancedarterialimagingandADCvaluewere86.9%and88.9%indifferentiatingMFCPand PC,whichwasbetterthananysinglemethod.Conclusion MRImulti-sequencecombinationisoneoftheeffectivemethodsforidentifyingPCand MFCP,andhasreferencevalueforclinicaldiagnosis.
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Objective To investigate the value of intravoxel incoherent motion (IVIM)and three-dimensional arterial spin labeling (3D-ASL)technique in preoperative grading diffuse gliomas.Methods Thirty-one patients with diffuse gliomas proved by pathology were enrolled,including 12 cases with low grade gliomas and 19 cases with high grade gliomas.All patients underwent 3D-ASL and IVIM examination before operation.Original dates were disposed in particular software,then the images of cerebral blood flow (CBF), D? and D were obtained,and ROI were selected and relevant parameter values were calculated.The parameter values were analyzed in SPSS,and unpaired two-tailed student t test were performed to compare parameter values between two groups.P<0.05 was regarded as statistically significant. ROC analysis curves were performed to acquire optimum threshold value,sensibility and specificity of each parameter.Finally,the sensibility and specificity of the associating screening of 3D-ASL and IVIM were obtained.Results The CBF and relative CBF(rCBF) values of high grade gliomas group [(103.89±27.00)mL/min?100 g and (4.28±0.63)mL/min?100 g]were significantly higher than those of low grade gliomas group [(63.96±22.17)mL/min?100 g and (2.72±0.84)mL/min?100 g];The D? and relative D?(rD?) values of high grade gliomas group [(3.82 ± 0.60 )mm2/s and 1.9 5 ± 0.30 ]were significantly higher than those of low grade gliomas group [(2.54±0.50)mm2/s and 1.28±0.14],the D and relative D (rD)values of high grade gliomas group [(0.58±0.12)mm2/s and 2.40±0.49]were significantly lower than those of low grade gliomas group [(0.75±0.12)mm2/s and 3.1 1±0.42](P<0.05). Statistical analysis in ROC demonstrated AUC value of 0.879 and 0.917 for CBF and rCBF,optimum threshold value of 79.24 mL/min?100 g and 2.85,sensibility of 84.2% and 100%,specificity of 83.3%and 83.3%;AUC value of 0.925,0.846,0.956 and 0.882 for D?,D ,rD? and rD ,optimum threshold value of 2.95 mm2/s, 0.63 mm2/s,1.40 and 2.95,sensibility of 94.7%,91.7%,94.7%and 75%,specificity of 83.3%,73.3%,91.7% and 94.7%.The sensibility and specificity of the associating screening of 3D-ASL and IVIM were 100% and 9 1.7%.Conclusion IVIM and 3D-ASL can be used to grade diffuse gliomas,and the values of rCBF,rD? and rD are optimal indicator.Combination of IVIM and 3D-ASL can improve the sensibility and specificity of grading diffuse gliomas.
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Zfyve16 (a.k.a. endofin or endosome-associated FYVE-domain protein), a member of the FYVE-domain protein family, is involved in endosomal trafficking and in TGF-β, BMP, and EGFR signaling. The FYVE protein SARA regulates the TGF-β signaling pathway by recruiting Smad2/3 and accelerating their phosphorylation, thereby altering their susceptibility to TGF-β-mediated T cell suppression. Zfyve16 binds to Smad4 and their binding affects the formation of Smad2/3-Smad4 complex in TGF-β signaling. However, the in vivo function of Zfyve16 remains unknown. In this study, we generated a Zfyve16 knockout mouse strain (Zfyve16) and examined its hematopoietic phenotypes and hematopoietic reconstruction ability. The proportion of Tcells in the peripheral blood of Zfyve16 mice increases compared with that in wild-type mice. This finding is consistent with the role of Zfyve16 in facilitating TGF-β signaling. Unpredictably, B cell proliferation is inhibited in Zfyve16 mice. The proliferation potential of Zfyve16 B-lymphoid cells also significantly decreases in vitro. These results suggest that Zfyve16 inhibits the proliferation of T cells, possibly through the TGF-β signaling, but upregulates the proliferation of B-lymphoid cells.
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Animales , Ratones , Linfocitos B , Metabolismo , Linfocitos T CD4-Positivos , Metabolismo , Movimiento Celular , Proliferación Celular , Genética , Péptidos y Proteínas de Señalización Intracelular , Genética , Metabolismo , Ratones Noqueados , Serina Endopeptidasas , Genética , Metabolismo , Transducción de Señal , Proteínas Smad Reguladas por Receptores , Metabolismo , Factor de Crecimiento Transformador beta , Metabolismo , Regulación hacia ArribaRESUMEN
We constructed recombinant adenoviruses expressing miR-29b2c (Ad-miR29b2c), and analyzed their effects on the proliferation and migration of HGC-27 and MGC-803 cells. miR-29b2c gene was amplified by PCR from genomic DNA and cloned into the pAdTrack-CMV vector to create the shuttle plasmid pAdT-29b2c. The recombinant plasmid was verified by restriction enzyme digestion and sequencing. The linearized shuttle vector was mixed with an adenoviral backbone plasmid (pAdEasy-1), followed by cotransformation into competent BJ5183 cells to generate the recombinant plasmid pAd-miR-29b2c. Finally, recombinant adenoviral vectors were generated by transfecting the recombinant plasmid into 293A packaging cell line. HGC-27 and MGC-803 cells were infected with the recombinant adenoviruses expressing pAd-miR-29b2c, then MTT and wound-healing assay were used to analyze the effects of pAd-miR-29b2c on the proliferation and migration of HGC-27 and MGC-803 cells. The miR-29b and miR-29c levels were significantly increased in HGC-27 cells after infected with pAd-miR-29b2c. MTT and wound-healing analysis also revealed a significant decrease in proliferation and migration of HGC-27 and MGC-803 cells compared to the control Ad-GFP-infected cells. Furthermore, western blotting results demonstrated that the protein expression level of δ-catenin was reduced in pAd-miR-29b2c transfected HGC-27 and MGC-803 cells. Taken together, the recombinant adenoviral vector was generated, and it can significantly inhibit the proliferation and migration of HGC-27 and MGC-803 cells.
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Objective To evaluate the diagnostic value of ESAT6 and CFP10 in smear-negative pulmonary tuberculosis.Methods 32 patients of smear-negative pulmonary tuberculosis were selected as the observation group.80 cases of other non-tuberculous respiratory diseases were selected as the control group.ESAT6 and CFP10 polypeptides were used as a stimulating source,the amount of IFN-γsecreted by peripheral blood T lymphocytes was measured by ELISPOT technique.The result was compared with serum anti-tuberculosis antibody (TB-Ab).Results The positive rates of ESAT-6,CFP-10 and combination detection in the observation group were 84.4%,78.1% and 90.6% respectively,which were significantly higher than those in the control group,the differences were statistically significant (x2 =71.94,67.43,73.51,all P < 0.01).The sensitivity (90.6%) and negative predictive value (96.1%) of ESAT6,CFP10 combined detection had no statistically significant differences compared with TB-Ab method (x2 =2.20,1.78,all P > 0.05).The specificity (92.5%),positive expected value (82.9%) and the diagnostic accuracy (92.0%) of ESAT6,CFP10 combined detection were significantly higher than those of TB-Ab (60.0%,40.7 %,62.5 %),the differences were statistically significant (x2 =11.24,12.07,7.92,all P < 0.01).The positive likelihood ratio of ESAT6,CFP10 combined detection was greater than 10 and negative likelihood ratio was less than 0.1.The possibility of diagnosis was significantly increased.Youden index co-detected by ESAT6 and CFP10 was 0.8,and that detected by TB-Ab was 0.3,indicated that the combined accuracy of ESAT6 and CFP10 was higher than TB-Ab.Conclusion Mycobacterium tuberculosis specific antigen ESAT6,CFP10 can be used as a stimulating source to stimulate effect T lymphocyte to secrete interferon-γ (IFN-γ).IFN-γ can be detected using ELISPOT.Mycobacterium tuberculosis specific antigen ESAT6,CFP10 can be used to quickly assistant the diagnosis of tuberculosis.They have higher sensitivity,specificity and good application value in the auxiliary diagnosis of smear-negative pulmonary tuberculosis.
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This paper was aimed to study the protective effects and related mechanisms of Zhen-Gan Xi-Feng (ZGXF) decoction containing serum on 6-OHDA-induced oxidative stress in PC12 cells.The ZGXF decoction containing rat serum with low-,medium-,and high-dose (8,16,or 32 g.kg-1) or blank serum was used to preprocess PC12 cells for 1 h,and cultured together with 100 μM 6-OHDA for 24 h.And then,cells were collected.The fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH-DA) and fluorescence microplate reader were used to detect the level of reactive oxygen species (ROS).Real-time quantitative PCR was used to analyze the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2),Nfe2l2,heme oxygenase-1 (HO-1),glutamate-cysteine ligase catalytic subunit (GCLc),and GCL modulatory subunit (GCLm).The luciferase report gene system was used to detect the antioxidant response element (ARE) activation.The results showed that ZGXF decoction-containing serum inhibited the 6-OHDA-induced oxidative stress,upregulated the Nfe2l2,HO-1 and GCLc mRNA expressions in cells processed with 6-OHDA.However,it has no significant effect on GCLm mRNA expression.It was concluded that ZGXF decoction-containing serum had protective effects on 6-OHDA-induced oxidative stress in PC 12 cells.Its mechanism may be correlated with the upregulation on Nfe2l2 mRNA expression,which activated ARE and further induced its downstream gene of phase Ⅱ detoxifying enzyme,as well as the HO-1 and GCLc mRNA expressions of antioxidant enzyme gene.
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Objective: Heart failure (HF) patients are usually associated with liver function impairment, Child-Turcotte-Pugh (CTP) scores can evaluate liver function, but its effect in HF patients has been unclear. We want to study the application of CTP scores in predicting the risk of death for in-hospital HF patients. Methods: A total of 1180 consecutive in-hospital HF patients were enrolled. According to CTP scores evaluated liver function at admission, the patients were divided into 3 groups: CTP grade A group, n=951, CTP grade B group, n=206 and CTP grade C group, n=23. The endpoint of this study was all-cause death. Results: There were 180 patients died at 1 year follow-up period, the in-hospital and 1 year mortalities were increased with the elevated CTP grades accordingly: for in-hospital mortalities in CTP grade A, B and C groups were (0.8%, 11.7% and 56.5%) respectively, P Conclusion: CTP scores may independently predict the risk of death for in-hospital HF patients, the levels of CTP scores might be used for evaluating the efficacy of in-hospital treatment.
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<p><b>OBJECTIVE</b>This study was designed to investigate the plasma level of soluble ST2 (sST2) and related influencing factors, and establish its reference value in the healthy community-based population in Beijing area of China.</p><p><b>METHODS</b>We measured plasma sST2 level by enzyme-linked immunosorbent assay between March 2012 and August 2012 in 1 334 healthy subjects in communities, including 597 males and 737 females. Empiric and quantile regression methods were used to determine the reference range of plasma sST2. A multiple linear regression model was established to analyze the factors that might affect the level of plasma sST2.</p><p><b>RESULTS</b>Gender is the most important factor affecting the plasma level of sST2 in healthy people. Plasma level of sST2 is significantly higher in men than in women (P < 0.01). Within each age strata, i.e. < 45, 45-54, 55-64, ≥ 65 years old, the plasma levels of sST2 were significantly higher in men than age-matched female (all P < 0.01). Age, body mass index (BMI), systolic blood pressure, diastolic blood pressure, and smoking did not affect plasma sST2 level. Reference range of sST2 was 5.7-53.5 µg/L for men and 4.4-42.4 µg/L for women (95% nonparametric reference interval). The one-side upper 95th percentile value of sST2 to discriminate the cardiovascular disease from healthy state was 47.2 µg/L for men and 37.2 µg/L for women.</p><p><b>CONCLUSIONS</b>This study established the normal reference range of plasma sST2 in healthy community-based population. The major influencing factor of sST2 level in healthy population is gender.</p>
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Femenino , Humanos , Masculino , Pueblo Asiatico , Enfermedades Cardiovasculares , China , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Análisis Multivariante , Valores de ReferenciaRESUMEN
<p><b>OBJECTIVE</b>To investigate the plasma level of amino-terminal pro-brain natriuretic peptide (NT-proBNP) and related influencing factors in a community-based healthy population in Beijing area.</p><p><b>METHODS</b>We measured plasma NT-proBNP level by fluoroimmunoassay between March 2012 and July 2012 from 1 034 healthy subjects (including 486 men and 548 women). Empiric method was used to determine the reference value and influencing factors were analyzed.</p><p><b>RESULTS</b>Age and gender are important factors affecting the level of NT-proBNP in healthy subjects. NT-proBNP plasma level is significantly higher in women than in men within each age strata below 75 years old, i.e. < 45, 45-54, 55-64 and 65-74 years old (P = 0.005, 0.001, 0.001, 0.011 respectively), but NT-proBNP plasma level is similar between male and female older than 75 years (P = 0.504). NT-proBNP level also increases with age irrespective of gender. Body mass index (BMI) is another independent influencing factor of NT-proBNP (P < 0.001), while estimated glomerular filtration rate is not influencing factor. The reference range of NT-proBNP is < 133 ng/L for men and < 289 ng/L for women aged < 55 years old, < 185 ng/L for men and < 333 ng/L for women aged between 55 and 64 years old, and < 465 ng/L for men and < 378 ng/L for women aged ≥ 75 years old.</p><p><b>CONCLUSION</b>The major influencing factors of NT-proBNP level in the healthy population are age, gender and BMI. It essential to establish normal reference range of NT-proBNP according to these factors for Chinese population.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pueblo Asiatico , Estado de Salud , Péptido Natriurético Encefálico , Fragmentos de Péptidos , Valores de ReferenciaRESUMEN
Objective To optimize the synthesis route of substituted quinazolinone and investigate the influence of tem-perature and time in the synthesis of key intermediates .Methods A parallel test was carried out to compare temperature ,time and the ester bond ammonolysis of condensing agent ,including the total analysis of experimental result .And orthogonal experi-mental design was used and the influence of temperature and time on the yield were investigated .Results and Conclusion The structure of the compound was confirmed by 1 H NMR ,MS and 13C NMR .When the reaction temperature was 30 ℃ ,the reac-tion time was 24 h with the condensing agent (DCC) and the reactant ratio of 1 .5∶1 ,the yield of ester bond ammonolysis was higher .The optimal preparation procedure of quinazolinone compounds was more available for industrial production .In vitro antifungal activity test results showed that the lead compounds measured on five clinical pathogenic fungi have the potential an-tifungal activity ,and are worth for further study .
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Objective:To observe the expression of Ki-67,P53 and LAT1 in tissues of esophageal squamous cell carcinoma and precancerous lesions and to investigate its clinical significance.Methods: Immunohistochemical method was used to detect the expressions of Ki-67,P53 and LAT1 in tissues from 20 cases of normal esophageal mucosa,68 cases of precancerous lesions including 21 cases of mild atypical hyperplasia,22 cases of atypical hyperplasia,25 cases of severe atypical hyperplasia and cancer tissues from 34 esophageal cancer patients.The relationship between the expression of the three in esophageal carcinoma was analysed.Results: Re-spectively,the positive expression rate of Ki-67 in normal esophageal mucosa, mild atypical hyperplasia, moderate dysplasia, severe dysplasia and carcinoma was 0%( 0/20 ) , 23.8%( 5/21 ) , 40.9%( 9/22 ) , 76.0%( 19/25 ) , 82.4%( 28/34 ) and the positive expression rate of P53 was 0%(0/20),14.3%(3/21),31.8%(7/22),48.0%(12/25),67.6%(23/34) and the positive expression rate of LAT1 was 0%( 0/20 ) , 19.0%( 4/21 ) , 36.4%( 8/22 ) , 52.0%( 13/25 ) , 76.5%( 26/34 ).The rank correlation analysis showed that the positive expression of Ki-67,P53 and LAT1 were significantly correlated with histological grade(r=0.626,0.427, 0.586,P<0.01) and the expression of Ki-67,P53 was positively correlated with LAT1 in esophageal carcinoma tissues(r=0.428, 0.596,P<0.01).Conclusion:Abnormal expression of Ki-67,P53,LAT1 protein was significant related to carcinogenesis of esophageal cancer and the combined detection of the three has important clinical significance.
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This study is to screen the Chinese herbal compounds which could inhibit the production of Abeta and investigate the underlying mechanism. Ten types of compounds which have potential value in the treatment of AD were selected as initial screening trial. The cell models which used could overexpress Abeta and beta-secretases or Abeta and gamma-secretases. Extracellular Abeta was determined by ELISA after the cell models treated with different concentrations of compounds (0.5-100 micromol x L(-1)), separately. Then the compounds were selected which could inhibit extracellular Abeta and their best concentration ranges were decided, too. Furthermore, the cell viability and apoptosis rate, the level of intracellular Abeta, beta and gamma-secretases were determined after the cell models treated with different concentrations of selected compounds. The results showed that 4 of the 10 compounds could reduce the level of extracellular Abeta; they were cryptotanshinone, astragalosides, gastrodin and paeoniflorin, and their best concentration ranges were 0.5-5.0, 0.5-5.0, 5.0-50, 1.0-25 micromol x L(-1), respectively. Further study indicated that the 4 selected compounds were nontoxic to the cellular models and lowering intracellular Abeta were more effective compared with extracellular; of which astragalosides and gastrodin showed dose-dependent inhibition to the activities of beta and gamma-secretases, with the maximum inhibiting rates of 78.2% and 80.3%, respectively. In conclusion, cryptotanshinone, astragalosides, gastrodin and paeoniflorin could inhibit the expression and secretion of Abeta, and the underlying inhibiting mechanism of astragalosides and gastrodin were related with the reduction of the beta and gamma-secretase activities, respectively.
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Five compounds, huyouyisu (1), vomifoliol (2), isoferulic acid (3), 1,2,3-trihydroxy-phenol (4) and p-hydroxy-phenol (5), were isolated from the peels of Citrus changshan-huyou Y.B.Chang for the first time by utilizing repeated column chromatography on silica gel. Among them, huyouyisu (1) is a new compound. The structure was elucidated by using 1D and 2D spectra.