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1.
J Clin Lab Anal ; : e23089, 2019 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-31709651

RESUMO

BACKGROUND: Trisomy 21 is a common aneuploid condition in humans and accounts for approximately one quarter of all aneuploid live births. To date, early diagnosis of Trisomy 21 remains a challenging task. Metabolomics may prove an innovative tool to study the early pathophysiology of Trisomy 21 at a functional level. METHODS: Ultra-performance liquid chromatography coupled with mass spectrometer (UPLC-MS) was used for untargeted metabolomic analysis of amniotic fluid samples from women having normal and trisomy 21 fetuses. RESULTS: Many significantly changed metabolites were identified between amniotic fluid samples from Trisomy 21 pregnancies and normal euploid pregnancies, such as generally lower levels of several steroid hormones and their derivatives, higher levels of glutathione catabolites coupled with lower levels of gamma-glutamyl amino acids, and increased levels of phospholipid catabolites, sugars, and dicarboxylic acids. The identification of a human milk oligosaccharide in amniotic fluid may worth further investigation, since confirmation of this observation may have significant implications for regulation of fetal development. CONCLUSIONS: The metabolisms in amniotic fluid from Trisomy 21 and normal pregnancies are quite different, and some of the significantly changed metabolites may be considered as candidates of early diagnostic biomarkers for Trisomy 21.

2.
Zootaxa ; 4691(5): zootaxa.4691.5.9, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31719383

RESUMO

A new species of stag beetle, Dorcus tianlongi Wang Zhou, new species (Coleoptera: Lucanidae: Lucaninae) is described from Guizhou Province, China. It is closely related to D. liyingbingi Huang Chen, 2013 and D. mencius (Kriesche, 1935). Diagnostic characters of the three species are illustrated and compared. Dorcus mencius is for the first time recorded from Henan Province of China.


Assuntos
Besouros , Animais , China
3.
Zootaxa ; 4668(2): zootaxa.4668.2.7, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31716631

RESUMO

A new species of flanged bombardier beetle form Hainan Island, China, Eustra yinggelingensis sp. nov. (Coleoptera, Carabidae, Paussinae, Ozaenini) is described. The new species was collected in rotten wood from Yinggeling Nature Reserve. Habitus and diagnostic features of the new species are illustrated. List, key and distribution map of all known Chinese Eustra species are given.


Assuntos
Besouros , Distribuição Animal , Animais , China , Ilhas
4.
Scand J Immunol ; 2019 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-31660620

RESUMO

Ulcerative colitis (UC) is an idiopathic, chronic inflammatory disease, which is characterized with overactive immune response. It is well established that the imbalance between Tregs and Th17 cells plays a pivotal role in pathogenesis of UC. In this study, we investigated the impact of functional changes in Treg subsets on Treg/Th17 ratio and further explored their clinical significance in the activity of UC. Treg subsets were comprehensively analysed using flow cytometry and in vitro cultured in both active and remission UC patients, of which nine active UC patients were further followed up. The correlation analyses were performed to explore the potential associations between Treg subsets and clinical indicators, as well as the impact of serum cytokines, detected by ELISA, on IL-17A secretion and CCR6 co-expression of Treg subsets. In active UC patients, we found CD45RA- FoxP3hi Tregs were obviously decreased and inversely correlated with disease activity, while CD45RA+ FoxP3lo Tregs were increased and positively correlated with disease activity. Meanwhile, IL-17A secretion and CCR6 co-expression levels in Tregs were significantly increased in active UC. Moreover, Tregs co-expressing CCR6 possesses higher level of IL-17A secretion. In nine followed up patients, we observed downregulated IL-17A secreting and CCR6 co-expression when achieving remission from active stage. In addition, IL-17A+ FoxP3+ and IL-17A+ FoxP3+ CCR6+ Tregs were positively correlated with serum IL-21 and disease activity, respectively. These findings suggested that upregulated IL-17A secretion and CCR6 co-expression in Treg subsets may be related to the imbalance between Tregs and Th17 cells and associated with the disease activity in UC patients.

5.
Proteomics Clin Appl ; : e1900075, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31579992

RESUMO

PURPOSE: Due to a lack of effective early diagnostic measures, new diagnostic methods for bacterial bloodstream infections (BSIs) are urgently needed. A protein-peptide profiling approach can be used to identify novel diagnostic biomarkers of BSIs. EXPERIMENTAL DESIGN: In this study, MALDI-TOF MS and nano-LC/ESI-MS/MS were used to analyze serum peptides. In addition, GO and network analyses were conducted as a means of analyzing these potential protein markers. Finally, the potential biomarkers were verified in independent clinical samples via ELISA. RESULTS: M/z 1533.8, 2794.3, 3597.3, 5007.3, and 7816.7 revealed an identical trend; the intensity of m/z 1533.8, 2794.3, and 3597.3 were higher in the infection group relative to controls, whereas the intensity of m/z 5007.3, and 7816.7 were lower in the infection group. Four peaks were successfully identified including ITIH4, KNG1, SAA2, and C3. GO and network analyses found these proteins to be involved in a myriad of biological processes, to form an interaction network, which may be correlated with BSI. ELISA results indicated that ITIH4, KNG1, and SAA2 were effective in differentiating infected from normal control group and the febrile group. CONCLUSIONS AND CLINICAL RELEVANCE: These biomarkers have the potential to offer new insights into the signaling networks underlying the development and progression of BSI. This article is protected by copyright. All rights reserved.

6.
Clin Chem Lab Med ; 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31639100

RESUMO

Introduction Element-tagged immunoassay coupled with inductively coupled plasma-mass spectrometry (ICP-MS) detection has the potential to revolutionize immunoassay analysis in clinical detection; however, a systematic evaluation with the standard guidelines of the assay is needed to ensure its performance meets the requirements of the clinical laboratory. Methods Carcinoembryonic antigen (CEA) was chosen for analysis using the proposed method. A systematic evaluation of the proposed assay was carried out according to the Clinical and Laboratory Standards Institute (CLSI). The 469 clinical samples were analyzed using the new method and compared with the electrochemiluminescent immunoassay (ECLIA) method. Results The measurement range of the assay was 1-900 ng/mL, with a detection limit of 0.83 ng/mL. The inter-assay and intra-assay imprecision were 4.67% and 5.38% with high concentration samples, and 9.27% and 17.64% with low concentration samples, respectively. The cross-reactivity (%) for different antigens was less than 0.05%, and the recovery was between 94% and 108%. Percentage deviation of all the dilutions was less than 12.5% during linearity estimation. The interference bias caused by different substances was less than 10%. The reference interval of the assay was 0-4.442 ng/mL. Comparison with the commercial ECLIA method for clinical sample detection, the proposed method showed a correlation of 0.9878 and no significant differences between the methods were observed (p = 0.6666). Conclusions The ICP-MS based immunoassay was successfully developed, and the analytical performance of the assay met the requirements of the CLSI, which fully proved the clinical transferability and application of the new method.

7.
Medicine (Baltimore) ; 98(38): e17315, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31568018

RESUMO

Early differential diagnosis of bloodstream infections (BSIs) caused by different sources and species of bacteria in hospitalized patients is crucial for the timely targeted interventions including appropriate use of antibiotics. The aim of this study was to identify 9 biomarkers for the early differentiation of gram-negative-bloodstream infection (GN-BSI), gram-positive (GP)-BSI, and fungal-BSI.A prospective study was conducted for a total of 390 inpatients who underwent blood culture in the Chinese PLA General Hospital from September 2015 to March 2018. Patients with positive culture of a single pathogen were divided into GN-BSI, GP-BSI, and Fungal-BSI groups, and a culture-negative disease control group was also established. The serum levels of macrophage inflammatory protein 1ß (MIP-1ß), tumor necrosis factor α (TNF-α), interleukin (IL)-3, interferon (IFN)-γ, IL-17A, IL-4, IL-12p70, and P-selectin were detected and the NLR was calculated from routine blood test. Receiver-operating characteristic analysis was used to determine the efficacy of various indicators in the differential diagnosis of BSIs. Prediction and validation experiments on clinical patient samples (263 cases) were also performed.The level of IL-3 in the GP-BSI group was significantly higher than those in the other 3 groups. The level of IFN-γ in the fungal-BSI group was significantly higher than those in the other 3 groups. NLR, MIP-1ß, TNF-α, IL-17A, and IL3 exhibited some efficacy when distinguishing between GN-BSI and GP-BSI and NLR had the largest area under curve (AUC) (0.728), followed by MIP-1ß with an AUC of 0.679. IFN-γ and IL-3 exhibited some value in differential diagnosis between GN-BSI and Fungal-BSI. IL-3, MIP-1ß, TNF-α, IFN-γ, NLR, IL-17A, and IL-4 exhibited some value in distinguishing fungal-BSI and GP-BSI, with IL-3 had the largest AUC (0.722), followed by MIP-1ß with an AUC of 0.703.NLR and MIP-1ß may be valuable in differentiating GN-BSI from GP-BSI in hospitalized patients. IFN-γ and IL-3 may be helpful in differential diagnosis GN-BSI and fungal-BSI. IL-3 and MIP-1ß exhibited some diagnostic efficacy in distinguishing fungal-BSI and GP-BSI. Additionally, IL-3 with high serum level may be a marker for GP-BSI and IFN-γ with high serum level may be a valuable marker for the prediction of Fungal-BSI. The utility of these biomarkers to predict BSIs owing to different pathogens in hospitalized patients needs to be assessed in further studies.


Assuntos
Bacteriemia/diagnóstico , Quimiocina CCL4/sangue , Infecção Hospitalar/diagnóstico , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Positivas/diagnóstico , Interferon gama/sangue , Interleucina-12/sangue , Interleucina-17/sangue , Interleucina-3/sangue , Interleucina-4/sangue , Micoses/diagnóstico , Proteínas NLR/sangue , Selectina-P/sangue , Fator de Necrose Tumoral alfa/sangue , Bacteriemia/sangue , Bacteriemia/microbiologia , Biomarcadores/sangue , Infecção Hospitalar/sangue , Infecção Hospitalar/microbiologia , Diagnóstico Diferencial , Feminino , Infecções por Bactérias Gram-Negativas/sangue , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Positivas/sangue , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Micoses/sangue , Micoses/microbiologia , Estudos Prospectivos
8.
J Bone Joint Surg Am ; 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31644522

RESUMO

BACKGROUND: Diagnosing periprosthetic joint infection (PJI) requires various laboratory and clinical criteria. The purpose of this study was to explore novel biomarkers that could rapidly diagnose PJI with high accuracy. METHODS: In this retrospective study of prospectively collected samples, 50 synovial fluid aspirates, 20 from the hip and 30 from the knee, were collected before surgery; 25 of the patients were diagnosed as having aseptic loosening (non-PJI) and 25, as having PJI according to the Musculoskeletal Infection Society criteria. A quadrupole orbital-trap mass spectrometry (MS) instrument was used to compare expression of proteins in patients with and without PJI. Proteins that were most efficacious for diagnosis of PJI were then determined using prediction analysis of microarray software and a random forest model. The most promising proteins were selected, and altered expression of these selected proteins was verified by ELISA (enzyme-linked immunosorbent assay) in an extended sample cohort. RESULTS: A total of 256 proteins were significantly upregulated (≥3.0-fold) and 14 proteins were downregulated in synovial fluid of patients with PJI compared with patients without PJI. The 3 most promising proteins were lactoferrin (LTF), polymorphonuclear leukocyte serine protease 3 (PRTN3), and myeloid nuclear differentiation antigen (MNDA). When MS was used for diagnosis of PJI, the area under the curve was 0.9888 for LTF, 0.9488 for PRTN3, and 0.9632 for MNDA. ELISA results verified that LTF, MNDA, and PRTN3 were sensitive, while LTF and MNDA were specific, for diagnosis of PJI. CONCLUSIONS: This proteomic study identified a previously noted protein and 2 novel candidate proteins as promising synovial fluid biomarkers for PJI diagnosis, and they should be further validated in future clinical trials. LEVEL OF EVIDENCE: Diagnostic Level III. See Instructions for Authors for a complete description of levels of evidence.

9.
Oncol Rep ; 2019 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-31524277

RESUMO

The present study aimed to investigate the effects of perfluorooctanoic acid (PFOA) on tumor cell migration, invasion and apoptosis by activation of the PI3K/AKT signaling pathway in human rhabdomyosarcoma (RMS). PFOA is a persistent, synthetic organic environment pollutant, which has been previously associated with multiple diseases, including cancer. The present study aimed to confirm whether PFOA can elicit cell growth in the RD subline of RBS. RD cells were treated with different concentrations of PFOA. Cell proliferation was evaluated using Cell Counting Kit­8 and cell cycle assays. Cell migration and invasion were determined using wound healing and Transwell assays. Apoptotic rates were estimated by Annexin V­FITC/propidium iodide staining. The expression levels of vimentin, serum/glucocorticoid­regulated kinase 1 (SGK1), cyclin E2, cyclin dependent kinase (CDK)2, p53, p21, p27, phosphatidylinositol­3 kinase (PI3K) and protein kinase B (AKT), and apoptosis­associated genes and proteins (including Bcl­2 and Bax) were detected by reverse transcription­PCR and western blot analyses. The results showed that PFOA significantly promoted RD cell proliferation, migration and invasion and significantly inhibited RD cell apoptosis. Exposure to PFOA also induced the expression of vimentin, SGK1, cyclin E2, CDK2, AKT, PI3K and Bcl­2, but suppressed the expression of Bax in the RD cells. The treatment of RD cells with BEZ235, a PI3K inhibitor, antagonized the effects of PFOA on metastatic formation and apoptosis. The results obtained show that the PI3K/AKT signaling pathway is implicated in mediating the pro­neoplastic effects of PFOA. The data suggests that PFOA is a carcinogen capable of promoting RD cell migration and invasion and inhibiting apoptosis through the PI3K/AKT signaling pathway.

10.
Biochem Biophys Res Commun ; 517(3): 439-444, 2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31376932

RESUMO

The clinical management of pediatric acute myeloid leukemia (AML) is still challenging and identification of drugs that can enhance the efficacy of standard of care is a potential therapeutic strategy. We show that pamidronate, a FDA-approved drug used for bone disorders, is an attractive candidate for AML treatment. Pamidronate inhibits proliferation and induces apoptosis of AML cells regardless of cellular and genetic heterogeneity. Pamidronate displays selective anti-AML activity by preferentially inhibiting survival and colony formation of AML CD34+ cells while normal bone marrow CD34+ cells are largely unaffected. Importantly, pamidronate remarkably enhances the inhibitory effects of all tested AML standard of care at subtoxic concentration. Mechanism studies show that pamidronate inhibits protein prenylation via dual action on geranylgeranylation and farnesylation, and subsequently decreases Ras activity. The rescue studies using overexpression of constitutively active Ras further confirm that pamidronate augments the efficacy of AML standard of care through inhibiting Ras. Since pamidronate is already used in clinic, our preclinical findings suggest that it may be an effective addition to treatment armamentarium for AML.

11.
Malar J ; 18(1): 262, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31366365

RESUMO

BACKGROUND: The Mindray BC-6800 haematology analyzer (BC-6800) provides a dedicated flag 'Infected RBC' (InR) and the number of InR (InR#)/the permillage of InR (InR‰) in routine blood testing as a screening tool for malaria in endemic areas. This study sought to evaluate the effectiveness of the BC-6800 flag parameter for aiding the diagnosis of malaria. METHODS: A total of 181 samples were tested using the Mindray BC-6800 haematology analyzer, including 117 malaria-infected samples collected from Yunnan, China, and 64 samples from healthy controls. Microscopy examination was conducted as reference when stained thick blood film revealed the presence of malaria parasites identified as Plasmodium vivax and Plasmodium falciparum. The receiver operating characteristic (ROC) curve analysis was developed using Analyse-it v4.92.3. The Kappa value was determined to evaluate the agreement between BC-6800 and light microscopy. RESULTS: The sensitivity of InR‰ generated by BC-6800 for P. vivax and P. falciparum was 88.3 and 24.1%, respectively; specificity of InR‰ for malaria parasites was 84.3 and 84.3%, respectively; positive predictive value and negative predictive value was 89.4 and 82.7% for P. vivax, and 52.8 and 60.3% for P. falciparum. There was a strong correlation between ΔWBC and InR‰ (R2 = 0.9731 for P. vivax and R2 = 0.9757 for P. falciparum). There was also a significant correlation between parasitaemia and InR# in P. vivax-infected samples (R2 = 0.734). InR# was evaluated using ROC curve analysis, the area under the ROC curve is 0.95 with a 95% confidence interval of 0.926 to 0.974, and the cut-off value is 0.01 × 109/L for P. vivax. However, the ring stage and the early trophozoite stage of Plasmodium cannot be detected easily on BC-6800, possibly because of the small size and low nucleic acid content of these stages. CONCLUSIONS: The findings suggest that the flag 'InR' and the parameters 'InR#/InR‰' provided by the BC-6800 haematology analyzer could be used to screen for malaria in a clinical setting.


Assuntos
Análise Química do Sangue/métodos , Sangue/parasitologia , Hematologia/métodos , Malária Falciparum/diagnóstico , Malária Vivax/diagnóstico , Plasmodium falciparum/isolamento & purificação , Plasmodium vivax/isolamento & purificação , Adolescente , Adulto , Idoso , Análise Química do Sangue/instrumentação , Criança , Pré-Escolar , China/epidemiologia , Feminino , Hematologia/instrumentação , Humanos , Malária Falciparum/epidemiologia , Malária Falciparum/parasitologia , Malária Vivax/epidemiologia , Malária Vivax/parasitologia , Masculino , Pessoa de Meia-Idade , Parasitemia/diagnóstico , Parasitemia/epidemiologia , Parasitemia/parasitologia , Prevalência , Curva ROC , Sensibilidade e Especificidade
12.
Clin Chim Acta ; 499: 34-40, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31469979

RESUMO

Mucopolysaccharindosis type II (MPS II) is a rare lysosomal storage disorder caused by deficient or absent activity of the iduronate-2-sulfatase (IDS) enzyme, which leads to pathological accumulation of the glycosaminoglycans(GAGs). The absence of early diagnosis can result in irreversible developmental, neurological, and physiological damage. The lack of clear understanding of the etiology of physiological dysfunction in MPS II has been a major obstacle to the development of new treatment. Therefore, a reliable biomarker for early diagnosis and exploration of pathogenic mechanism are of great importance. Proteomics provides powerful tool for protein expression alterations and study of complicated pathological process. This study was performed to identify the differential protein profile in urine of MPS II patients using two-dimensional gel electrophoresis(2D-PAGE)combining with MALDI-TOF/TOF and a total of 15 differentially expressed proteins were identified. Content of alpha1-antitrypsin, Gm2 activator and lipocalin-type prostaglandin D synthase was measured by ELISA method. The value of urinary α1-AT/Cr in MPS II group was 0.79 ±â€¯0.10 mg/mmol, significantly higher than 0.42 ±â€¯0.05 mg/mmol in healthy control group; whereas the value of GM2A/Cr and L-PGDS/Cr in MPS II group was 1.30 ±â€¯0.12 µg/mmol and 9.86 ±â€¯1.16 ng/mmol respectively, which was significantly lower than 2.19 ±â€¯0.19 µg/mmol and 13.98 ±â€¯1.48 ng/mmol in healthy control group. The proteins can be considered as accessory diagnostic biomarkers for MPS II. This approach helped to discover early diagnostic markers and provided a better understanding of the pathogenic mechanism of MPS II.

13.
Medicine (Baltimore) ; 98(30): e16570, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31348286

RESUMO

BACKGROUND: Perioperative bleeding during total knee arthroplasty (TKA) is an ongoing problem for surgeons. Intravenous or topical application of tranexamic acid (TXA) can effectively stop bleeding, but there is still no uniform standard for the best method of administration and dose. METHODS: From October 2016 to September 2018, 218 patients with unilateral primary knee osteoarthritis requiring knee replacement were enrolled and randomly divided into four groups. Group 1 (n = 55) received intra-articular injection (IAI) of TXA and peri-articular injection (PAI) of placebo, group 2 (n = 55) received IAI of placebo and PAI of TXA, group 3 (n = 51) received IAI of TXA and PAI of TXA, and group 4 (n = 57) received double placebo (IAI of placebo and PAI of placebo). The demographic characteristics, surgical indices, hematological indices, wound healing history, and thromboembolic events were investigated. RESULTS: Eight patients were lost to follow-up and 210 patients were included in the analysis. The median TBLs in patients who received IAI of TXA and PAI of placebo and those who received IAI of placebo and PAI of TXA were 470.81 ml and 481.54 ml, respectively. These TBL levels were significantly higher compared to those in patients who received IAI of TXA and PAI of TXA (359.18 ml, P ≤ .001), but significantly lower compared to those in patients who received the double placebo (522.71 ml, P ≤ .001). Compared to other groups, more patients in the double placebo group needed a blood transfusion (P = .013). In the short-term, the double placebo group had higher VAS pain scores and less ROM after surgery (P = .011 and P = .001, respectively). In the long-term (6-month follow-up), there were no significant differences in ROM, VAS, DVT, PE, or wound-related complications. CONCLUSION: The combined use of IAI and PAI of TXA can significantly reduce the TBL and the need for blood transfusion without delaying wound healing or increasing the risk of DVT and PE. In the short-term after surgery, this combined method reduces the pain VAS scores and improves the ROM; however, there are no long-term effects on VAS and ROM.


Assuntos
Antifibrinolíticos/uso terapêutico , Artroplastia do Joelho/métodos , Perda Sanguínea Cirúrgica/prevenção & controle , Ácido Tranexâmico/uso terapêutico , Fatores Etários , Idoso , Antifibrinolíticos/administração & dosagem , Transfusão de Sangue/estatística & dados numéricos , Índice de Massa Corporal , Relação Dose-Resposta a Droga , Método Duplo-Cego , Vias de Administração de Medicamentos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/cirurgia , Fatores Sexuais , Fatores Socioeconômicos , Tromboembolia/epidemiologia , Cicatrização/efeitos dos fármacos
14.
Clin Appl Thromb Hemost ; 25: 1076029619863492, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31311294

RESUMO

Pregnancy is a hypercoagulable state associated with an increased risk of venous thrombosis. Calibrated automated thrombogram (CAT) is a test to monitor the thrombin generation (TG), a laboratory marker of thrombosis risk, and increases during normal pregnancy, but it is still unclear whether TG is related to the use of insulin in pregnant women with gestational diabetes mellitus (GDM). We performed thrombin generation by CAT on 135 normal pregnant women, including 43 in first trimester, 32 in second trimester, 60 in third trimester, respectively; 68 pregnant women with GDM were also enrolled, 19 patients with GDM using insulin to control blood glucose and 49 patients control their blood glucose through diet and exercise with noninsulin treatment. The overall CAT parameters were calculated using descriptive statistics method with mean ± standard deviation. Mean endogenous thrombin potential, peak thrombin generation, and StartTail time increased significantly with the pregnancy. There was no significant difference in TG test parameters except StartTail time(P = .003) in insulin-treated GDM group when compared to those without insulin in the GDM group. The normal ranges for CAT parameters in pregnant women were determined. Thrombin generation increased significantly in first trimester and remains stable in second and third trimester. The use of insulin in patient with GDM did not affect thrombin generation test. Our study helps to establish the reference range of thrombin generation in Chinese normal pregnant population and provide more basis to predict the risk of thrombus complicating during pregnancy.

15.
J Glob Antimicrob Resist ; 19: 358-364, 2019 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-31216492

RESUMO

OBJECTIVES: The aim of this study was to perform a detailed genomic characterisation of IncR plasmids from China. METHODS: Three IncR plasmids (p13190-tetA, p02085-tetA and p30860-tetA) from clinical isolates ofKlebsiella pneumoniae, Citrobacter freundii and Enterobacter cloacae, respectively, were fully sequenced using high-throughput genome sequencing and were compared with five previously sequenced IncR plasmids (pHN84KPC, pSH-01, pK245, pKPC_P16 and pKPC-LK30) from China. RESULTS: The eight IncR plasmids from China possessed conserved IncR backbones composed of repB, parAB, umuCD, retA and resD. Resistance accessory modules integrated into the IncR backbones included multidrug resistance (MDR) regions in p30860-tetA, p02085-tetA, p13190-tetA and pK245, blaKPC-2 regions in pHN84KPC, pKPC-LK30 and pKPC_P16, and the ΔTn1721-sil region in pSH-01. These resistance accessory modules were inserted at a site between retA and vagD, resulting in loss of the backbone genes vagCD in some of the plasmids. The resistance accessory modules differed dramatically from one another and carried distinct profiles of resistance markers. In particular, all of p13190-tetA, p02085-tetA, p30860-tetA, pHN84KPC, pSH-01 and pK245 carried tetracycline resistance tet gene modules, and the carbapenemase gene blaKPC-2 was identified in pHN84KPC, pKPC-LK30 and pKPC_P16. In addition, one or more regions responsible for plasmid replication and/or maintenance were found in some of the resistance accessory modules, facilitating stable replication of corresponding IncR plasmids at steady-state copy numbers. CONCLUSIONS: This detailed comparative genomics analysis of IncR plasmids from China provides a deeper insight into the diversification and evolution of IncR plasmids.

16.
Clin Lab ; 65(6)2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31232024

RESUMO

BACKGROUND: The Pentra MS CRP hematology analyzer (hereinafter the Pentra analyzer) can simultaneously provide 5-part leukocyte differential and C-reactive protein (CRP). The aim of the study was to investigate the performance of CRP determination by the Pentra analyzer. METHODS: The precision, limit of quantitation (LoQ), carryover, linearity, stability, and comparability of the Pentra analyzer were determined. The Passing-Bablok regression analysis and the Bland-Altman graphs illustrated the correlation for CRP concentration analyzed by the Pentra analyzer and BN-II analyzer. RESULTS: The within-run precision of CRP determination by the Pentra analyzer had a CV < 2.0% in peripheral blood, which met the requirements of the instructions (CV ≤ 10%). The Pentra analyzer had a total CV of 5.35% and 5.52% at a CRP concentration of 4.1 and 80 mg/L, respectively. The LoQ value for the Pentra analyzer was 0.96 mg/L. The carryover was 0.57% for peripheral blood and 0.86% for plasma by the analyzer. The stability of CRP results was good, when the anticoagulation samples were stored at room temperature or 4°C within 48 hours (deviation < 5%). The linearity range for whole blood samples was 0 - 188.13 mg/L (r² = 0.9992). There was high correlation of the CRP results analyzed with the Pentra analyzer and BN II analyzer. The Passing-Bablok regression analysis and the Bland-Altman graphs showed the bias plot display excellent agreement between the two assays (the mean value for the Pentra 2.19 mg/L and the BN-II 2.35 mg/L, n = 101). CONCLUSIONS: The results of CRP determination by the Pentra analyzer have the advantages of accuracy and reliability, and it is suitable for routine use in emergency laboratory and small to medium-size laboratories.

17.
J Cardiothorac Vasc Anesth ; 33(10): 2685-2694, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31064730

RESUMO

OBJECTIVE: Reducing mortality is a key target in critical care and perioperative medicine. The authors aimed to identify all nonsurgical interventions (drugs, techniques, strategies) shown by randomized trials to increase mortality in these clinical settings. DESIGN: A systematic review of the literature followed by a consensus-based voting process. SETTING: A web-based international consensus conference. PARTICIPANTS: Two hundred fifty-one physicians from 46 countries. INTERVENTIONS: The authors performed a systematic literature search and identified all randomized controlled trials (RCTs) showing a significant increase in unadjusted landmark mortality among surgical or critically ill patients. The authors reviewed such studies during a meeting by a core group of experts. Studies selected after such review advanced to web-based voting by clinicians in relation to agreement, clinical practice, and willingness to include each intervention in international guidelines. MEASUREMENTS AND MAIN RESULTS: The authors selected 12 RCTs dealing with 12 interventions increasing mortality: diaspirin-crosslinked hemoglobin (92% of agreement among web voters), overfeeding, nitric oxide synthase inhibitor in septic shock, human growth hormone, thyroxin in acute kidney injury, intravenous salbutamol in acute respiratory distress syndrome, plasma-derived protein C concentrate, aprotinin in high-risk cardiac surgery, cysteine prodrug, hypothermia in meningitis, methylprednisolone in traumatic brain injury, and albumin in traumatic brain injury (72% of agreement). Overall, a high consistency (ranging from 80% to 90%) between agreement and clinical practice was observed. CONCLUSION: The authors identified 12 clinical interventions showing increased mortality supported by randomized controlled trials with nonconflicting evidence, and wide agreement upon clinicians on a global scale.

18.
Clin Lab ; 65(5)2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31115220

RESUMO

BACKGROUND: CYP2D6*10 is mainly responsible for the large pharmacokinetic variability of routinely administered metoprolol in middle-aged and elderly Asian patients. Utilizing an efficient method for identifying the CYP2D6*10 genotypes is clinically important for evaluating the pharmacokinetic effect of administration of metoprolol. This study attempted to evaluate the effectiveness of the two methods used to detect the rs1065852 and rs1135840 SNPs of the CYP2D6*10 gene. METHODS: Blood samples were processed for the collection of genomic DNA from 198 subjects across Chinese population, and detection of CYP2D6*10 (rs1065852 and rs1135840) was performed using the PyroMark Q24 pyrose-quencing and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS). The discordant results were further validated with Sanger sequencing. We eventually attempted to assess some features of these two methods including reliability, rapidness, being appropriate, and cost-effectiveness. RESULTS: Genotyping of rs1065852 and rs1135840 detected by MALDI-TOF MS were concordant with those identified by PyroMark Q24 pyrosequencing in all 198 (100%) individuals. The hands-on-time and the turnaround time were shorter in the PyroMark Q24 pyrosequencing method than in the MALDI-TOF MS method for SNP of CYP2D6*10. In terms of being cost-effective and high-throughput, the MALDI-TOF MS method outperformed the PyroMark Q24 pyrosequencing method. CONCLUSIONS: CYP2D6*10 genotypes detected by PyroMark Q24 pyrosequencing and MALDI-TOF-MS showed that both methods were reliable, rapid, appropriate, and cost-effective methods. These methods are valuable for clinical applications.

19.
PLoS One ; 14(5): e0217298, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31125378

RESUMO

OBJECTIVE: The aim of this study is to evaluate the performance of different platelet counting methods (optical, impedance, fluorescence and hand counting) applied in different analysers by comparing with the international flow cytometric reference method (IRM). METHODS: A total of 333 blood samples from different subgroups (168 cases with thrombocytopenia, 136 cases with normal platelet counts and 29 cases with thrombocytosis) were tested. Regarding IRM as the gold standard, we compared the accuracy and precision of different platelet count methods; i.e. LH780 (impedance), BC-6000 Plus (optical (O) and impedance (I)), Sysmex XN-9000 (optical (O), impedance (I), fluorescence (F)), and hand counting. RESULTS: Sysmex XN-9000-F (r = 0.988) had the best correlation with IRM for thrombocytopenic samples; BC-6000 Plus-I (r = 0.966) was more relevant to IRM than any other method for samples with normal platelet counts. Correlation between Sysmex XN-9000-I (r = 0.960) and IRM was the highest among these methods for samples with thrombocytosis. For bias evaluation, the average bias of Sysmex XN-9000-F was -1.5 × 109/L (95% LA = -9.4 to +6.4) for samples with thrombocytopenia, compared with IRM. BC-6000 Plus-I had a small mean difference with IRM for samples with normal platelet counts or thrombocytosis. Moreover, all evaluated methods had acceptable sensitivity, specificity, and concordance rates as compared with IRM in the diagnosis of thrombocytopenia and thrombocytosis. CONCLUSIONS: Platelet counting by Sysmex XN-9000-F is more accurate than other methods for thrombocytopenic samples. BC-6000 Plus-I has superior association and consistency for normal platelet counts. As for thrombocytosis patients, Sysmex XN-9000-I has the highest correlation with IRM while Sysmex XN-9000-O has the highest diagnosis efficacy.

20.
Clin Lab ; 65(4)2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30969092

RESUMO

BACKGROUND: The Mindray BC-6800 automated hematology analyzer is an automated hematology analyzer and 5-part leukocyte differential counter for in vitro diagnostic use in clinical laboratories. It is necessary to undergo an evaluation before the instrument is used to test patient samples. METHODS: The performance was evaluated with regards to precision, linearity, carry-over, and method comparison. The flag performances were evaluated and compared with the Sysmex XE-2100 hematology analyzer and manual microscope in the hematology laboratory of a tertiary hospital in China. RESULTS: There was minimal carryover (< 0.05%) and excellent linearity for white blood cells and platelet (PLT) counts (r > 0.999). The BC-6800 displayed very good correlation (r > 0.97) with the XE-2100 for blood cell count and cell differential parameters. In a comparison of 295 leukocyte differential count results analyzed in parallel with manual microscopy, the main flags (immature granulocytes, blasts, abnormal lymphocytes) showed approxi-mately the same sensitivity and specificity on both analyzers (sensitivity > 90%, specificity > 78%). CONCLUSIONS: The BC-6800 showed excellent performance and supplied confidence in flag information for abnormal samples in the routine hematology laboratory.


Assuntos
Química Clínica/instrumentação , Química Clínica/métodos , Hematologia/instrumentação , Hematologia/métodos , Automação , Automação Laboratorial , Contagem de Células Sanguíneas/instrumentação , China , Humanos , Contagem de Leucócitos , Leucócitos/citologia , Modelos Lineares , Microscopia , Contagem de Plaquetas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
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