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1.
Background
New high‐performance liquid chromatography (HPLC) method was developed for the determination of vitamin K1 and two forms of vitamin K2 (MK‐4 and MK‐7) in human serum, and the levels of vitamin K were determined in 350 samples of postmenopausal women.Methods
Vitamin K was determined by HPLC with fluorescence detection after postcolumn zinc reduction. The detection was performed at 246 nm (excitation) and 430 nm (emission). The internal standard and 2 mL of ethanol were added to 500 μL of serum. The mixture was extracted with 4 mL of hexane, and solid phase extraction was then used.Results
The HLPC method was fully validated. The intra‐ and interday accuracy and precision were evaluated on two QC samples by multiple analysis, and CV were less than 10%. The limit of quantification for MK‐4 was found at 0.04 ng/mL, for K1 0.03 ng/mL, and for MK‐7 0.03 ng/mL. The mean recoveries of the corresponding compounds were 98%‐110%. Serum levels of MK‐4, K1, and MK‐7 in postmenopausal women with osteoporosis were 0.890 ± 0.291 ng/mL, 0.433 ± 0.394 ng/mL, and 1.002 ± 1.020 ng/mL, respectively (mean ± SD). Serum levels of MK‐4, K1, and MK‐7 in postmenopausal women without osteoporosis were 0.825 ± 0.266 ng/mL, 0.493 ± 0.399 ng/mL, and 1.186 ± 1.076 ng/mL, respectively (mean ± SD).Conclusion
New HPLC method for the determination of vitamins K1, MK‐4, and MK‐7 in serum was evaluated and validated. This method is highly specific and sensitive with the low limit of quantification.2.
Background
To establish reference intervals of carbohydrate antigen 19‐9(CA 19‐9) according to the CLSI CA28‐A3 guideline and to evaluate age‐ and gender‐related variations.Methods
Serum CA 19‐9 values of 10 149 healthy subjects (from 20 years old to 60 years old) were measured from location health checkups. The relationship between CA 19‐9 and age was analyzed using Spearman's approach. The reference intervals of CA19‐9 were established using Q2.5 and Q97.5, and the 90% confidence intervals of upper limits were calculated.Results
The reference intervals of CA 19‐9 were 1.98‐25.12 U/mL for males (1.97‐25.06 U/mL for 20‐50 years old and 2.31‐26.13 U/mL for 50‐60 years old) and 2.36‐29.29 U/mL for adult (20‐60 years old) females. The upper limit of reference intervals for all individuals was 26.45 U/mL; the level of CA 19‐9 is higher in females than males. Carbohydrate antigen (CA) 19‐9 is significantly associated with aging in adult males(r = .0930, P < .0001), but not in females (P = .4734).Conclusions
Establishing reference intervals for CA19‐9 and giving age‐related reference intervals of CA19‐9 using a big data of healthy adult, we first discovered that CA19‐9 tends to increase with age in adult males but not in females.3.
Background
Therapeutic monitoring of tacrolimus is essential for reducing organ rejection and adverse effects. The measurement of tacrolimus in whole blood is taken by many automated platforms. We evaluated the analytical performance of the Dimension TAC assay, which is an upgraded reagent from the previous Dimension TACR assay.Methods
The evaluations involved determination of precision, linearity, detection capability, and reagent lot‐to‐lot variability between three lot numbers. Correlation studies were conducted using the Dimension TACR assay, Architect, Elecsys assay, and MassTrak LC‐MS/MS.Results
The total coefficient of variation was below 10%. Acceptable linearity was observed in their respective reportable ranges. The limit of blank, limit of detection, and limit of quantification were 0.29, 0.47, and 0.81 ng/mL, respectively. Correlation analysis indicated that the Dimension TAC assay results were comparable to that of the Dimension TACR assay, Architect, and Elecsys results in liver and heart transplant patients. In kidney transplant patients, the Dimension TAC assay showed the poor correlation with Architect and Elecsys. The results from these assays were slightly higher than that of MassTrak. We found little lot‐to‐lot reagent variation among the reagents evaluated.Conclusion
The overall analytical performance of the Dimension TAC assay is acceptable for therapeutic monitoring in clinical practice. Our study that compared different platforms may provide some useful information regarding which test method to use.4.
Background
To improve the accuracy of the routine methods in laboratory medicine, ion chromatography with a simple sample treatment procedure, which can completely remove the proteins and/or organics in human serum, has been developed for the determination of serum cations.Methods
Chromatographic conditions for the separate and simultaneous determination of K, Na, Ca, and Mg were investigated. Furthermore, various factors influencing the mineralization of human serum, such as the selection and amount of oxidant, were also examined systematically and optimized.Results
The optimized experimental conditions are as follows: 1.0 mL of serum specimen digested with 2 mL nitric acid (120°C) followed by 2 mL hydrogen peroxide (80°C). The specimens were then redissolved and determined by ion chromatography under the optimum eluent concentration of 32 mmol/L methanesulfonic acids. The measurement accuracy and precision are less than 1.0% for all the analytes by analyzing NIST certified reference materials, IFCC‐RELA specimens and serum specimens. The results were also comparable with the reference values obtained by the inductively coupled plasma mass spectrometry (ICP‐MS), which were found to be in good agreement.Conclusions
Ion chromatography with a simple sample treatment procedure for the determination of cations in human serum with high sensitivity and specificity was developed. The proposed method could be recommended as a candidate reference method for the determination of serum cations.5.
Background
Ongoing efforts in the development of HBsAg detection kits are focused on improving sensitivity and specificity. The purpose of this study was to evaluate an improved, highly sensitive quantitative assay, “Lumipulse HBsAg‐HQ”, a chemiluminescent enzyme immunoassay designed for a fully automated instrument, the “Lumipulse G1200”.Methods
Serum samples for reproducibility, dilution, correlation, sensitivity, and specificity studies were obtained from patients at the Osaka University Hospital. Seroconversion and sensitivity panels were purchased from a commercial vender. Subtype, sensitivity panels, and HBsAg recombinant proteins with one or two amino acid substitutions were prepared in‐house.Results
The coefficients of variation for the low, medium, and high concentration samples ranged from 1.93 to 2.55%. The HBsAg‐HQ reagent for dilution testing showed good linearity in the 0.005‐150 HBsAg IU/mL range and no prozone phenomenon. All 102 HBV carrier samples were positive by HBsAg‐HQ, while other commercial reagents showed one or more to be negative. In the seroconversion panel, the 14‐day blood sample was positive. The sensitivity against HBsAg‐HQ “ad” and “ay” subtypes was 0.025 ng/mL. Comparisons among the HBsAg‐HQ, HISCL, and Architect HBsAg reagents were performed using the Bland‐Altman plot. Specificity for 1000 seronegative individuals was 99.7%. HBsAg‐HQ detected 29 positive serum among 12 231 routinely obtained serum samples, which showed concentrations of 0.005‐0.05 HBsAg IU/mL.Conclusions
According to these results, the Lumipulse HBsAg‐HQ assay, with a highly sensitive limit of detection of 0.005 IU/mL, may facilitate the development of a better management strategy for a considerable proportion of infected patients.6.
Background
Dysfibrinogenemia is a rare coagulation disorder caused by mutations in the fibrinogen gene that results in abnormal fibrinogen function. Dysfibrinogenemia has a wide spectrum of clinical manifestations including asymptomatic(55%), hemorrhage (25%), and thrombosis (20%).Methods
We reported a 30‐year‐old woman with 35 weeks gestation. She was misdiagnosed with hypofibrinogenemia in a local hospital, and then she was treated with fibrinogen concentrate. However, she was diagnosed as dysfibrinogenemia in our hospital base on her low function fibrinogen level (0.55 g/L) and her normal immunologic fibrinogen level (3.80 g/L). This patient had neither bleeding symptom nor thromboembolic event. Her obstetrical history included one normal pregnancy in 2008 with uneventful full‐term delivery.Results
Multidisciplinary experts suggested that there should be no specific intervention in this case because of the patient had no previous episodes of abnormal bleeding or thrombotic. She had an uneventful delivery with no abnormal bleeding symptom or thromboembolic.Conclusion
Dysfibrinogenemia patients without personal or family history of bleeding and thromboembolic events, do not need specific therapeutic intervention.7.
Background
Among cardiovascular diseases (CVD), acute coronary syndrome (ACS) is the main manifestation, corresponding to signs and symptoms that occur with ischemia and outcome of angina or acute myocardial infarction (AMI). The aim of this study was to investigate the performance of biochemical markers eligible in a chest pain protocol, using Point of care Test (POCT), in patients in a reference emergency room.Methods
In this study, 1380 medical records of patients of both genders were evaluated, ranked by applying chest pain protocol using the Manchester Triage System (MTS). Markers for myocardial injury were measured in serial analysis including myoglobin (Mgb), creatine kinase MB fraction mass (CK‐MB), and cardiac troponin I (cTnI).Results
Acute myocardial infarction was predominant in males (P < .001), in patients with hypertension (P < .001), and in those with previous myocardial infarction (P < .026) and significant electrocardiogram (ECG) data for AMI screening (P < .001). A multivariate regression model showed as predictors for AMI the variables ECG data by admittance at the emergency room, previous AMI history, levels of both Mgb at the third hour, and cTnI at the sixth hour after admission.Conclusion
This study showed the importance of a rapid and serial test as a cardiac marker for AMI screening, as well as has indicated the importance of time between the onset of chest pain and admission to the emergency room as an efficient aid in diagnosing this life‐threatening disease.8.
Background
Spontaneous bacterial peritonitis (SBP) is frequently occurring infection among patients with liver cirrhosis, defined by polymorphonuclear (PMN) leukocytic count ≥250 cell/mm3 with or without a positive ascitic fluid (AF) bacterial culture . So, this study aimed to investigate the diagnostic value of flow cytometry versus manual counting of ascitic fluid PMNL in cirrhotic patients, with clinical suspicion of SBP.Methods
A hospital‐based cross‐sectional study was carried out on 320 cirrhotic patients with clinical suspicion of SBP . Abdominal paracentesis was performed in all cases for microscopic manual and flow cytometry counting of PMNL. Anti‐HLA‐DR, anti‐CD15, anti‐CD16, and anti‐CD45 monoclonal antibodies were used for flow cytometry method.Results
Flow cytometric PMNL count had 100% sensitivity and specificity, while manual PMNL count had a sensitivity of 65.52% and specificity of 90% with significant difference (P value < .05).Conclusion
Flow cytometry is more reliable rapid method for PMNL counting, than the manual method that is less accurate and time‐consuming in diagnosing clinically suspected SBP.9.
Purpose
Endothelial damages are one of the most important causes of persistent hypertensive complications. The aim of our study was to discuss the relationship between the molecular markers of endothelial damage, thrombomodulin, and complications in hypertension.Methods
A total 132 cases of hypertensive patients, including 13 patients with damage of target organs, were selected as research subjects. And grouping was based on different levels of blood pressure. The blood pressure and thrombomodulin levels were detected among all cases, and their drinking, smoking, and other medical records were tracked.Results
Higher plasma concentration of thrombomodulin was demonstrated in subjects with hypertensive complications compared with without complications [24.5 (18.1,37.55) vs 12.1 (9.1,22.3) TU/mL, P = .001, respectively]. The optimum thrombomodulin cutoff value was determined to be more than 15.5 TU/mL, with a sensitivity of 92.3% and a specificity of 63%. With the increase in blood pressure level, thrombomodulin levels in three groups gradually raised [6.15(5.475,12.75) vs 9.75(7.725,13.35) vs 16.45 (10.125,23.725) TU/mL, P = .007, respectively].Conclusion
With the increase in blood pressure and the occurrence of complications, thrombomodulin showed an increasing trend, which was caused by an increase in the degree of endothelial injury. So, thrombomodulin may serve as a clinically meaningful marker of the progression of hypertension.10.
Background
To establish maternal thyroid‐stimulating hormone (TSH) reference ranges for first trimester screening from 11 + 0 to 13 + 6 weeks of gestation.Methods
A total of 10 592 singleton and 201 twin consecutive Caucasian pregnant women who underwent simultaneously prenatal first trimester Down's syndrome screening and thyroid function screening from January 2010 to November 2017 were included in the study. Women with positive antithyroid peroxidase antibody (TPOAb) and positive personal history of thyroid disease were previously excluded. TSH was measured by immunochemiluminescent assay on ci 16200 Abbott Architect analyzer. Nonparametric percentile method (also known as CLSI C28.A3) was used for the determination of reference ranges.Results
We established reference ranges of TSH for the period of gestation from 11 + 0 to 13 + 6 weeks of pregnancy as 0.16‐3.43 mU/L for singleton Caucasian pregnancies and 0.02‐2.95 mU/L for twin Caucasian pregnancies. The median (IQR) of TSH for singleton pregnancies was higher than that for twin pregnancies (1.25 mU/L (0.83‐1.81) vs 0.84 (0.37‐1.47), respectively; P < .0001).Conclusions
Each first trimester screening center should be aware of which type of immunoassay their laboratory uses. TSH reference ranges in women during the first trimester of pregnancy are lower than those for general population. Twin pregnancies have lower TSH than singleton pregnancies.11.
Objective
The prevalence of nonalcoholic fatty liver disease (NAFLD) has been rapidly increased, becoming a public health problem worldwide. Our objective was to investigate the association between urine retinol‐binding protein (RBP) and NAFLD in a Chinese population and develop a multivariate logistic regression model for NAFLD prediction.Methods
A total of 317 NAFLD patients and 391 healthy controls were enrolled in this cross‐sectional study based on inclusion and exclusion criteria, from whom fasting urine and blood were collected for further study. Urine RBP level and other parameters were measured and compared between NAFLD subjects and controls.Results
Urine RBP levels (expressed by RBP/creatinine ratio) in NAFLD patients were significantly higher than controls (median 133.1 mg/g vs 110.7 mg/g; P < .001). Urine RBP/creatinine ratio was verified as an independent factor for NAFLD prediction after adjustment in multivariate logistic regression. The area under curve (AUC) of receiver operating characteristic (ROC) was 0.889 with the 95% confidence interval from 0.867 to 0.912.With a cutoff point of 0.215, the sensitivity and specificity of urine RBP/creatinine ratio in NAFLD prediction were 81.1% and 84.5%, respectively.Conclusion
Our results demonstrated that urine RBP/creatinine ratio was an independent risk factor for NAFLD while the predictive model for NAFLD diagnosis is noninvasive with high sensitivity and specificity.12.
Background
The inflammatory response to Mycobacterium tuberculosis bacilli influences tuberculosis (TB) progression. In this study, we aimed to identify the Phe206Leu polymorphism and serum L‐selectin level in TB patients, compared to healthy individuals.Methods
Ninety patients with a diagnosis of TB and 90 healthy controls were selected in this study. The serum L‐selectin level was determined, using ELISA. L‐selectin polymorphism was also evaluated using PCR. For data analysis, SPSS was used at a significance level of 0.05.Results
According to the findings, the mean±SD age of the participants was 57.5 ± 18.4 and 56.5 ± 17.5 years in the TB and healthy groups, respectively. The TB group showed a significantly higher serum L‐selectin level (1721.1 ± 330.9) versus the healthy controls (1624 ± 279). The L‐selectin Phe allele frequencies were higher than the Leu allele frequencies in the main population, whereas the patients and controls were not significantly different. Eight (0.04%) subjects had Leu/Leu genotypes, 84 (46.6%) carried Phe/Leu genotypes, and 88 (48.8%) had Phe/Phe genotypes. Our results showed that the groups were not significantly different regarding L‐selectin genotypes.Conclusion
TB patients had a significantly higher serum L‐selectin level, compared to the controls. Based on the findings, the incidence of TB and L‐selectin polymorphism in the Phe206Leu gene had no significant association.13.
Background
Cellular and brain metabolism of dopamine can be correlated with a number of neurodegenerative disorders, our study was to explore a simple and efficient method to detect dopamine in real samples.Methods
A new quantum dots (CdTe QDs) could be prepared using the hydrothermal method, the electrochemical biosensor was established by dropping CdTe QDs on the surface of glassy carbon electrode (GCE).Results
The CdTe QDs/GCE exhibited the excellent electrochemical catalytic activity toward dopamine (DA) with good stability and high sensitivity in presence of interfering substances. The detection limit of DA was calculated by differential pulse voltammetry (DPV) as low as 0.3 μmol L−1 with a linear dynamic range of 1 μmol L−1 to 400 μmol L−1.Conclusion
In this paper, the proposed electrochemical biosensor could be effectively used for the direct and rapid detection of DA in human serum and urine samples.14.
Background
Recent studies have found circular RNAs (circRNAs) involved in the biological process of cancers. However, little is known about their functional roles in glioblastoma.Methods
Human circRNA microarray analysis was performed to screen the expression profile of circRNAs in IDH1 wild‐type glioblastoma tissue. The expression of hsa_circ_0008344 in glioblastoma and normal brain samples was quantified by qRT‐PCR. Functional experiments were performed to investigate the biological functions of hsa_circ_0008344, including MTT assay, colony formation assay, transwell assay, and cell apoptosis assay.Results
CircRNA microarray revealed a total of 417 abnormally expressed circRNAs (>1.5‐fold, P < .05) in glioblastoma tissue compared with the adjacent normal brain. Hsa_circ_0008344, among the top differentially expressed circRNAs, was significantly upregulated in IDH1 wild‐type glioblastoma. Further in vitro studies showed that knockdown of hsa_circ_0008344 suppressed glioblastoma cell proliferation, colony formation, migration, and invasion, but increased cell apoptotic rate.Conclusions
Hsa_circ_0008344 is upregulated in glioblastoma and may contribute to the progression of this malignancy.15.
Background
Use of total laboratory automation (TLA) system has expanded to microbiology and hemostasis and upgraded to second and third generations. We herein report the first successful upgrades and fusion of different versions of the TLA system, thus improving laboratory turnaround time (TAT).Methods
A 21‐day schedule was planned from the time of pre‐meeting to installation and clinical sample application. We analyzed the monthly TAT in each menu, distribution of the “out of range for acceptable TAT” samples, and “prolonged time out of acceptable TAT,” before and after the upgrade and fusion.Results
We installed and customized hardware, middleware, and software. The one‐way CliniLog 2.0 version track, 50.0‐m long, was changed to a 23.2‐m long one‐way 2.0 version and an 18.7‐m long two‐way 4.0 version. The monthly TAT in the outpatient samples, before and after upgrading the TLA system, were uniformly satisfactory in the chemistry and viral marker menus. However, in the tumor marker menu, the target TAT (98.0% of samples ≤60 minutes) was not satisfied during the familiarization period. There was no significant difference in the proportion of “out of acceptable TAT” samples, before and after the TLA system upgrades (7.4 ‰ and 8.5 ‰) . However, the mean “prolonged time out of acceptable TAT” in the chemistry samples was significantly shortened to 17.4 (±24.0) minutes after the fusion, from 34.5 (±43.4) minutes.Conclusions
Despite experimental challenges, a fusion of the TLA system shortened the “prolonged time out of acceptable TAT,” indicating a distribution change in overall TAT.16.
Background
Syntax score (SS), which is an angiographic tool used in grading the complexity of coronary artery disease (CAD), has prognostic importance in coronary artery disease (CAD) and provides important information regarding selection of revascularization strategy. C‐reactive protein (CRP) and albumin are indicators of inflammation, and high levels of them are associated with high SS. We aimed to investigate whether baseline CRP to albumin ratio C‐Reactive Protein/Albumin Ratio (CAR), an easily available and novel inflammatory marker, is associated with SS.Method
A total 403 consecutive patients with stabile angina pectoris, who underwent coronary angiography for suspected CAD from January 2015 to June 2016, were classified into two groups, low SS (≤22) and intermediate‐high SS (>22).Results
C‐Reactive Protein/Albumin Ratio was significantly higher in patients with intermediate‐high SS group (P < .001). In multivariate regression analysis, CAR remained an independent predictor of intermediate‐high SS group together with hypertension and LDL. The predictive performance of CAR, CRP, and albumin was compared by ROC curve analysis. CAR surpassed CRP and albumin in predicting intermediate‐high SS group. CAR >6.3 predicted an intermediate‐high SS with sensitivity and specificity of 86.8% and 43.4%, respectively.Conclusion
C‐Reactive Protein/Albumin Ratio was more tightly associated with the complexity and severity of CAD than CRP and albumin alone and was found to be an independent predictor for intermediate‐high SS group.17.
Background
Nephrolithiasis is a worldwide health problem that affects almost all populations. This study aimed to evaluate the association between rs12654812 of regulator of G protein signaling 14 (RGS14) gene and nephrolithiasis in the Chinese population.Methods
A total of 1541 participators including 830 cases and 711 controls were included from Guangxi area in China. Age, sex, BMI, smoking status, drinking status, creatinine, uric acid, and urea nitrogen were analyzed between the case group and control group.Results
We found that the G/A+A/A genotypes of rs12654812 had a significantly increased nephrolithiasis risk after adjusting age, sex, BMI, smoking, drinking, and hypertension, compared with G/G genotype (OR = 1.361, 95% CI = 1.033‐1.794, P = .029). This hazardous effect was more pronounced in subgroup of age < 50, ever smoking, ever drinking, creatinine normal, and high uric acid. The G/A genotype of rs12654812 also had a significantly increased nephrolithiasis risk compared with G/G genotype. The A allele of rs12654812 significantly increased the risk of nephrolithiasis compared with the G allele after adjusting for age, sex, BMI, smoking, drinking and hypertension (OR = 1.277, 95% CI = 1.013‐1.609, P = .038).Conclusions
Our results suggest that the RGS14 polymorphism is involved in the etiology of nephrolithiasis and thus may be a genetic marker for nephrolithiasis.18.
Backround
The relationship between maternal zinc level and birth weight, birth week, delivery type, garvida, maternal age, etc., contribute to diagnosis and clinical follow‐up.Method
Multivariate investigated for data of 275 patients were obtained during their pregnancy periods until birth. 3 cc blood samples were centrifuged for 15 minutes at 2500 g within a period of 30 minutes and were stored at −80°C until the time of analysis. The zinc levels of the patients were found to be within the range of 49‐129 μg/dL. Patients were divided into 8 groups according to their zinc levels (49‐59, 60‐69, …, 120‐129) and the relationships of zinc level with the parameters related to the mode of delivery, week of delivery, birth weight, age, early membrane rupture, live‐stillbirth, and gravid were statistically analyzed to determine differences between the groups.Results
There was a significant difference between the live births and stillbirths with a 95% confidence level regarding the zinc level. The zinc level affected the live‐stillbirth status; patients with a zinc level of 49‐59 μg/dL had stillbirths, the live birth rate for 59‐69 μg/dL was approximately 50%, whereas it was approximately 88% for in the patients with a zinc level of 109‐119 μg/dL. All patients with a zinc level of 119 μg/dL and above had live births.Conclusion
Based on the results of this study, it is suggested that zinc supplementation may be an appropriate treatment for the pregnant women with low zinc levels to provide the realization of live births .19.
Background
The von Willebrand factor (VWF) multimer test is required to correctly subtype qualitative type 2 von Willebrand disease (VWD). The current VWF multimer assays are difficult, nonstandardized, and time‐consuming. The purpose of this study was to evaluate the clinical utility of the commercial VWF multimer kit by Sebia (Lisses, France), an electrophoresis technique yielding same‐day results.Methods
Ten healthy volunteer plasma samples, in‐house reference plasma (IRP) and commercial normal plasma (CNP) samples, 10 plasma samples from patients with a known VWD type, 1 hemophilia A plasma sample, and 7 external quality assurance (EQA) samples were analyzed using the commercial VWF multimer kit. Additional coagulation testing included measurements of VWF antigen (VWF:Ag), VWF activity (VWF:Ac), and FVIII activity (FVIII:C).Results
The CNP results revealed a relative loss of the highest molecular weight multimers; therefore, IRP was preferred as the reference sample. The interpretations of 10 patients with a known VWD type could be successfully reproduced and agreed with previous VWF multimer results. In all EQA surveys, the multimer results and final VWD diagnosis agreed with expert opinion.Conclusions
The VWF multimer assay by Sebia is easy to perform and can be successfully implemented in any clinical laboratory for second‐stage evaluation of VWD. The resolution power of multimer distribution is adequate to correctly classify VWD types 1, 2A, 2B, and 3.20.