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BACKGROUND: It is not yet clear whether the trace blood gas analyzer can be used for biochemical detection of newborns. This study aimed to evaluate the reliability of the method for the detection of bilirubin in infants. METHODS: Based on the Clinical and Laboratory Standards Institute (CLSI) EP15-A2 document, the analytical performance of the blood gas analyzer method for bilirubin detection in neonates was validated. The resulting data of 363 simultaneous bilirubin detection with blood gas analyzer (optical method) and biochemical analyzer (enzymatic method) were reviewed. According to the CLSI EP9-A3 document, the relevance and consistency of the measurement results were evaluated by Pearson correlation analysis, Passing-Bablok regression, and Bland-Altman deviation analysis. RESULTS: The precision and accuracy of the Werfen GEM 4000 blood gas analyzer for the detection of different levels of bilirubin samples adhered to the manufacturer's statement and industry quality standards. The bilirubin detection values of the 2 methods showed a good correlation, and both of them were significantly correlated (P300 μmol/L group. CONCLUSIONS: The method for detecting neonatal total bilirubin by trace blood gas analyzer basically meets the clinical requirements and can be used for the preliminary screening of neonatal jaundice. However, for severe hyperbilirubinemia that requires close monitoring of dynamics, a precise enzymatic quantification is required.

Objective. To assess KIF11 expression in human thyroid tumor tissues and further evaluate its involvement in thyroid cancer. Methods. The expression of KIF11 in 71 cases of thyroid carcinoma as well as corresponding tissues was detected by the immunohistochemical (IHC) method. Patients were divided into the high KIF11 expression as well as low expression groups based on the staining levels. In addition, to study the relationship between the expression of KIF11 as well as clinicopathological features, the effects of KIF11 were detected on the proliferation, apoptosis, and cell cycle of two types of thyroid cancer cells, TPC-1 and KTC-1, through colony formation assays, MTT assays, and FCM assays, respectively. We further assessed the potential effects of KIF11 on tumor growth using an animal model. Results. The significantly high expression of KIF11 in thyroid tumor tissues was revealed, and the correlations between KIF11 expression levels as well as clinical pathological features (T stage and intraglandular dissemination) of patients were revealed. We further noticed that KIF11 knockdown remarkably suppressed thyroid cancer cell proliferation as well as induced cell apoptosis of thyroid cancer cells. Additionally, KIF11 contributed to tumor growth of thyroid cancer cells in mice. Conclusions. We noticed the involvement of KIF11 in the progression of thyroid cancer.

Hemolysis is one of the most frequent causes of pre-analytical errors in the emergency department (ED), and it can lead to inaccurate blood results and often requires repeat testing. The purpose of the present study was to evaluate the effects of true hemolysis occurring in ED blood samples on routine clinical biochemistry tests using the VITROS(®) 5600 Integrated system. A total of 92 pairs of blood samples were collected from 92 ED patients. Each pair of samples included one hemolyzed sample and one successful (non-hemolyzed) redraw from the same patient. A total of 21 common laboratory analytes and the hemolytic index (HI) were examined. The degree of hemolysis (slight, mild, moderate and heavy) was determined based on the HI. A clinically significant difference in one analyte was defined as a difference greater than its Clinical Laboratory Improvement Amendments of 1988 (CLIA'88) total allowable error (TAE) limits. The results demonstrated that the mean differences in 11 serum analytes (unconjugated bilirubin, Ca(2+), equivalent CO(2), Cl(-), creatinine, glucose, Mg(2+), phosphorus, Na(+), urea nitrogen and uric acid) in hemolyzed and non-hemolyzed samples were within their CLIA'88 TAE limits, while the differences in the other 10 analytes [alanine aminotransferase (ALT), albumin (ALB), amylase (AMYL), aspartate aminotransferase (AST), total bilirubin (TBIL), creatine kinase (CK), CK-myocardial band isoenzyme (CK-MB), lactate dehydrogenase (LDH), K(+) and total protein (TP)] in paired samples in at least one of the four groups were greater than their CLIA'88 TAE limits. These results suggest that hemolysis had a notable impact on ALT, ALB, AMYL, AST, TBIL, CK, CK-MB, LDH, K(+) and TP levels. Furthermore, for ALT, AMYL, TBIL and TP, wet chemistry methods displayed superior anti-hemolytic ability compared with dry chemistry methods. Notably, a high concentration of AST was less affected by hemolysis.

Background/aims To evaluate cellular immune function in systemic lupus erythematosus (SLE) patients over 60 years old, the association between antinuclear antibody (ANA) titers and the ratio of CD4+ /CD8+ was analyzed in this study. The distribution of ANAs and extractable nuclear antibodies (ENAs) in a healthy elderly population was also investigated. Methods Serum ANA titers were assayed by indirect immunofluorescence (IIF) and the CD4+ /CD8+ T-cell ratio was determined by flow cytometry in 76 SLE patients and 30 healthy control individuals. IIF of cytoplasm and nuclear and nucleolar staining were performed on samples taken from 286 healthy elderly individuals. ENA levels were determined using a strip enzyme immunoassay among patients that tested positive for ANAs. Results ANA titers were negative in the 30 control individuals, but were positive in the 76 SLE patients. Based on ANA titers, the SLE patients were stratified to low (≤ 1:320), medium (1:640 to 1:1,280), and high (≥ 1:2,560) titer groups. The average CD4+ /CD8+ ratio of the SLE group was significantly lower than that of the control group. Among the 286 healthy elderly volunteers, 59 (20.63%) tested positive for ANAs. A homogeneous pattern was present in 47.46% of those 59 patients and a granule pattern in the karyoplasm was present in 33.90%. Furthermore, of the 59 patients, ENAs immunoassay was positive in 18 (30.51%); Sjogren syndrome-related antigen A (SSA)/52 kd and Sjogren syndrome antigen B (SSB)/La were the two major antibodies. Conclusion The significantly lower CD4+ /CD8+ ratio among SLE patients over 60 years old is associated with deregulated immune responses and the development of SLE. A low ANA titer (1:160) is common in healthy elders, emphasizing the importance of considering age when determining if the evaluation of ANA titers is to be included in autoimmune disease diagnosis.

The determination of specific IgE (sIgE) level is of great importance in IgE-mediated food allergies. Our aim was to develop a homogeneous immunoassay—light-initiated chemiluminescent assay (LICA)—for measuring allergen sIgE of a single component in egg white, thus evaluating the LICA-sIgE assay as a useful tool in the diagnosis of food allergy. The LICA-sIgE assay was performed by incubating serum sample with anti-human IgE antibody coated with chemiluminescer beads, streptavidin-coated sensitizer beads, and biotinylated antigens, which consist of four components in egg white. Serum samples from egg allergic patients (n = 70) and healthy volunteers (n = 30) were collected. For calibration, purified human IgE was used as the calibrator. Working conditions of this homogeneous immunoassay were optimized, analytical performance was determined, and correlation of the results between LICA and ImmunoCAP was evaluated. The assays were performed in 8-well plates with a sample volume diluted to 1:10 of 25 μl. Intra-assay precision (% coefficient of variation) ranged from 1.83 to 4.13%, and inter-assay precision ranged from 2.70 to 8.70%. It exhibited excellent sensitivity, which could distinguish between positive samples and negative samples even at a large dilution level. The sIgE-LICA and ImmunoCAP correlated well in patients allergic to single component (r 2 = 0.929). Also, the components ovomucoid and ovalbumin were best at predicting ImmunoCAP results, with the same area under the ROC curve (AUC) of 0.81, and a specificity of 90.0 and 93.3%, respectively. Our data show effective performance characteristics of LICA to detect sIgE in human serum based on component-resolved diagnostic tests (CRD). The homogeneous sIgE-LICA assay has the following key advantages: requires no washing, simplicity and rapidity, reproducibility, high-throughput, good performance in a liquid phase assay, and good suitability for sIgE diagnosis in food allergy based on CRD.

Background The long non-coding RNAs (lncRNAs) are emerging as important regulators in cancer progression. Clear cell renal cell carcinoma (ccRCC) is one of the most common urological cancers with poor prognosis. In this study, we examined the functional role of the lncRNA, nuclear enriched abundant transcript 1 (NEAT1) in ccRCC progression. Methods We performed quantitative real time PCR and western blotting assays to measure mRNA and protein expression levels, respectively. CCK-8 assay, cell invasion and migration assays were used to determine the cell proliferative, cell invasive and migratory ability. Flow cytometric analysis was performed to examine cell apoptosis. Results The expression levels of NEAT1 was up-regulated in ccRCC tissues and up-regulation of NETA1 was positively correlated with tumor size, higher Fuhrman grade, and lymph node metastasis, and also predicts worse 5-year survival rate of patients with ccRCC. NEAT1 knock-down by NEAT siRNAs transfection suppressed cell proliferation and induced cell apoptosis in ccRCC cell lines. In addition, NEAT1 knock-down suppressed cell invasion and migration and inhibited the mRNA and protein expression levels of epithelial-mesenchymal transition-related markers in ccRCC cell lines. Conclusions In conclusion, NEAT1 may be an important mediator in the regulation of ccRCC progression and predicts the poor prognosis in patients with ccRCC.

Quantification of testosterone serves an important role in the differential diagnosis of androgen‑related endocrine diseases. Mass spectrometry exhibits higher accuracy and lower variability than immunoassays, especially at low testosterone concentrations. The present study developed and validated an isotope dilution ultra‑performance liquid chromatography tandem mass spectrometry method for determination of human serum testosterone. The serum was equilibrated with an isotopic internal standard and treated with acidic buffer to release hormones from their binding proteins. Testosterone was extracted via two serial liquid‑liquid extractions. In the first stage, the lipid fractions from an acidic buffer solution were isolated using ethyl acetate and n‑hexane. The organic phase was evaporated and reconstituted in a basic buffer solution. In the second stage, the polar impurities of n‑hexane extraction were removed. Total testosterone in serum was quantified via ultra‑performance liquid chromatography tandem mass spectrometry in multiple reaction monitoring mode with positive electrospray ionization. The coefficient of variation of the method for intra‑ and inter‑assay was 2.13% (1.40‑2.77%) and 3.44% (3.06‑3.66%), respectively. The recovery ranged from 94.32 to 108.60% for different samples. The limit of detection was 0.50 ng/dl and the linear range was from 1.00 to 1,000.00 ng/dl. In addition, the extraction efficiency in three different levels of quality control of the serum ranged from 85.02 to 93.29%. Moreover, structural analogues were investigated and were not indicated to affect the quantification of testosterone. The present method may enable quantification of testosterone in a clinical setting with high precision and accuracy.

Long noncoding RNAs (lncRNAs) are emerging as important modulators in the biological processes and tumorigenesis. However, whether lncRNAs are involved in endometrial endometrioid adenocarcinoma (EEC) remains unclear. In the present study, we explored the expression pattern, clinical significance and biological function of nuclear enriched abundant transcript 1 (NEAT1) in EEC. The expression levels of NEAT1 were elevated in EEC tissues and cell lines, and higher expression levels of NEAT1 were positively correlated with FIGO stage and lymph node metastasis. Overexpression of NEAT1 in HEC-59 cells transfected with pGCMV-NEAT1 promotes cell growth, colony formation ability as well as invasive and migratory ability; while knock-down of NEAT1 in HEC-59 cells by siNEAT1 transfection exhibited the opposite effects. Flow cytometry analysis showed that overexpression of NEAT1 led to an increase in S-phase cells and attenuated cell apoptosis, and knock-down of NEAT1 induced G0/G1 arrest and also induced cell apoptosis in HEC-59 cells. Tumor metastasis real-time PRC array showed that six metastasis-related genes (c-myc, insulin like growth factor 1(IGF1), matrix metallopeptidase 2 (MMP-2) and matrix metallopeptidase 7(MMP-7) were up-regulated, and Cadherin 1 and TIMP metallopeptidase inhibitor 2 were down-regulated) in NEAT1-overexpressing HEC-59 cells. Further qRT-PCR and western blot results confirmed that c-myc, IFG1, MMP-2 and MMP-7 were dys-regulated by NEAT1. Together, our data underscore the significance of NEAT1 in EEC development, and NEAT1 may a potential therapeutic target for EEC.

Contrast-induced acute kidney injury (CI-AKI) is a serious complication of diagnostic coronary angiograph and percutaneous coronary intervention (PCI). However, the exact pathophysiological mechanisms underlying CI-AKI development are largely unknown. The present study examined whether urinary semaphorin 3A levels predict the development of CI-AKI in patients undergoing PCI. This study enrolled 168 patients with stable angina undergoing elective PCI. Serial urine samples, obtained at baseline and 2, 6, 12, 24, 36, and 48 h post-PCI were analyzed by semaphorin 3A and neutrophil gelatinase-associated lipocalin (NGAL) ELISA kit. AKI was defined as an increase in serum creatinine beyond 50% according to the RIFLE classification system. Receiver operator characteristic (ROC) curve analyses identified optimal semaphorin 3A and NGAL values for diagnosing CI-AKI. CI-AKI occurred in 20 of 168 patients. There were no significant differences in the baseline clinical characteristics and angiographic findings between non-AKI patients group and AKI patients group. Both urinary semaphorin 3A and NGAL levels significantly increased at 2 and 6 h post-PCI. ROC analysis showed that the cut-off value of 389.5 pg/mg semaphorin 3A at 2 h post-PCI corresponds to 94% sensitivity and 75% specificity and the cut-off value of 94.4 ng/mg NGAL at 2 h post-PCI corresponds to 74% sensitivity and 82% specificity. Logistic regression showed that semaphorin 3A levels at 2 and 6 h post-PCI were the significant predictors of AKI in our cohort. Urinary semaphorin 3A may be a promising early biomarker for predicting CI-AKI in patients undergoing PCI.

// Yixin Zhang 1, 2, * , Xiaoteng Cui 3, * , Li Ning 1 and Dianjun Wei 1 1 Department of Clinical Laboratory, The Second Hospital of Tianjin Medical University, Tianjin 300211, PR China 2 School of Medical Laboratory, Tianjin Medical University, Tianjin 300070, PR China 3 Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, PR China * These authors have contributed equally to this work Correspondence to: Dianjun Wei, email: weidianjun01@163.com Keywords: tumor necrosis factor-α; single nucleotide polymorphisms; sepsis; rs1800629; rs361525 Received: September 16, 2017 Accepted: November 13, 2017 Published: November 30, 2017 ABSTRACT This meta-analysis of 23 eligible articles comprehensively and quantitatively evaluated the effects of tumor necrosis factor-α ( TNF-α ) rs1800629 and rs361525 polymorphisms on sepsis risk. We found that TNF-α rs1800629 was associated with increased sepsis risk in the overall population in four genetic models, including A vs. G ( P 1). TNF-α rs361525 was also associated with increased sepsis risk in Asian patients in the four genetic models (all P 1). Begg’s and Egger’s tests excluded large publication bias, and sensitivity analysis indicated stable results. Our results suggest that the G/A genotype of TNF-α rs1800629 and rs361525 increases sepsis risk in an Asian population.

Persistent low-grade inflammation is thought to underlie the pathogenesis of many chronic diseases, such as cardiovascular diseases and metabolic syndrome. Autoimmunity is correlated with increased levels of chronic low-grade inflammation, and immunoglobulin M (IgM) is reactive to autoantigens and believed to be important for autoimmunity. Triglyceride (TG) is fatty acid carrier and initiator of oxidative stress, and it has been hypothesized that TG stimulates B cells to secrete IgM. However, few studies have investigated the relationship between TG and IgM in human populations. We designed a cross-sectional and prospective cohort study to evaluate how serum TG levels are related to IgM concentration. Participants were recruited from Tianjin Medical University General Hospital-Health Management Centre. Both a baseline cross-sectional (n = 10,808) and a prospective assessment (n = 2,615) were performed. Analysis of covariance was used in the cross-sectional analysis. After multiple adjustments for confounding factors, serum IgM level in the highest quartile of TG in males was significantly higher than levels in lower quartiles (P

Silver nanoparticles are increasingly recognized for their utility in biological applications, especially antibacterial effects. Herein, we confirmed the antibacterial effect of silver nanoparticles on Escherichia coli using the conventional optical density (OD) and colony-forming units (CFU) method and used flow cytometry (FC), TEM and BrdU ELISA to investigate the mechanisms of this effect. From the results, we conclude that AgNPs can simultaneously induce apoptosis and inhibit new DNA synthesis in the cells in a positive concentration-dependent manner. This study presents the first induction of apoptosis in these bacteria by AgNPs in this field. Our findings may provide a new strategy for the use of silver nanoparticles in antibacterial applications.