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BACKGROUND: This study investigated the performance of the MC-100i, a pre-commercial digital morphology analyzer utilizing a convolutional neural network algorithm, in a multicentric setting involving up to 11 tertiary hospitals in China. METHODS: Blood smears were analyzed by MC-100i, verified by morphologists, and manually differentiated. The classification performance on WBCs and RBCs was evaluated by comparing the classification results using different methods. The PLT and PLT clump counting performance was also assessed. The total assay time including hands-on time was evaluated. RESULTS: The agreements between pre- and post-classification were high for normal WBCs (κ > 0.96) and lower for overall abnormal WBCs (κ = 0.90). The post-classification results correlated well with manual differentials for both normal and abnormal WBCs (r > 0.93), except for basophils (r = 0.8480) and atypical lymphocytes (r = 0.8211). The clinical sensitivity and specificity of each RBC abnormality after verification were above 90 % using microscopy reviews as the reference. The PLTs counted by the MC-100i before and after verification correlated well with those measured by the PLT-O mode (r = 0.98). Moreover, PLT clumps were successfully classified by the analyzer in EDTA-dependent pseudothrombocytopenia blood samples. CONCLUSIONS: The MC-100i is an accurate and reliable digital cell morphology analyzer, offering another intelligent option for hematology laboratories.
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Hematología , Leucocitos , Humanos , Centros de Atención Terciaria , Eritrocitos , China , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: IgA nephropathy (IgAN) is the most frequently occurring primary glomerulonephritis. A lack of specific biomarkers hinders the early diagnosis and treatment of this disease. This study analyzes and validates potential serum biomarkers using mass spectrometry proteomics. METHODS: Global proteomics profiles of serum from 60 patients with IgAN and 43 healthy control subjects were compared to identify significantly changed proteins. These proteins were validated with targeted proteomics using parallel reaction monitoring (PRM) in an independent validation set consisting of samples from 67 different stage IgAN patients and 60 healthy controls. RESULTS: A total of 37 significantly changed proteins were found in the discovery set, among which 18 proteins were identified as potential biomarkers for IgAN through PRM assays in the validation set. Of these 18 proteins, IgGFc-binding protein, MS-A1 light chain variable region, transthyretin, ficolin-3, and myosin-reactive immunoglobulin light chain variable region were up-regulated in different IgAN stages, B cell receptor heavy chain variable region, rheumatoid factor RF-ET6, heavy chain Fab, cryocrystalglobulin CC1 heavy chain variable region, FLJ94213, lumican, and Q68CN4 (uncharacterized protein) were down-regulated in different IgAN stages. These proteins support previous findings that CKD is accompanied by altered immune response. CONCLUSIONS: This study lays the groundwork for additional research using biomarkers to clinically diagnose IgAN. These proteins are potential molecular markers that could help us understand the potential molecular mechanism of IgAN.
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Glomerulonefritis por IGA , Insuficiencia Renal Crónica , Humanos , Cromatografía Liquida , Proteómica/métodos , Espectrometría de Masas en Tándem , Glomerulonefritis por IGA/diagnóstico , Inmunoglobulina A , BiomarcadoresRESUMEN
BACKGROUND: Neuromyelitis optica spectrum disorders (NMOSD) is an autoimmune demyelinating disease, leading recurrently relapses and severe disability. There is a need for new biomarkers to meet clinical needs in diagnosis and monitoring. METHODS: Through liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis, brain lesions from NMO animal models were analyzed to identify potential biomarkers. Then, we assessed the levels of serum glial fibrillary acidic protein (sGFAP), neurofilament light chain (sNfL), Tau protein (sTau) and Ubiquitin C-terminal hydrolase L1 (sUCHL1) using an ultrasensitive single molecule array (Simoa) of AQP4-IgG + NMOSD patients, myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) patients, multiple sclerosis (MS) patients and healthy controls (HCs). Additionally, we further explored the early diagnosis value of these proteins. RESULTS: There were 72 differentially expressed proteins between the NMO and control groups. NfL abundance was elevated when GFAP, UCHL1, and Tau abundance was decreased in the NMO group. Then, we observed that the sGFAP and sUCHL1 levels in patients with NMOSD in the early stage were significantly increased compared to those in control participants. Combined ROCs of the sGFAP, sNfL, and sUCHL1 levels to better predict NMOSD with relapse stages was optimal. Notably, univariate and multivariate analyses demonstrated that the sGFAP and sNfL levels were higher in patients with brain lesions, while the sUCHL1 levels were higher in those with spinal cord lesions during recent relapse. CONCLUSIONS: These findings suggested that sGFAP, sNfL, and sUCHL1 displayed good diagnostic performance in AQP4-IgG + NMOSD and could be novel candidates for early discrimination.
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Neuromielitis Óptica , Animales , Neuromielitis Óptica/diagnóstico , Acuaporina 4/análisis , Cromatografía Liquida , Espectrometría de Masas en Tándem , Biomarcadores , Autoanticuerpos , Inmunoglobulina G , Glicoproteína Mielina-OligodendrócitoRESUMEN
BACKGROUND: Neoadjuvant chemoradiotherapy (nCRT) prior to surgery is the standard treatment for patients with locally advanced rectal cancer (LARC), while parts of them show poor therapeutic response accompanied by therapy adverse effects. Predictive biomarkers for nCRT response could facilitate the guidance on treatment decisions but are still insufficient until now, which limits the clinical applications of nCRT in LARC patients. METHODS: In our study, 37 formalin-fixed paraffin-embedded tumor biopsies were obtained from patients with LARC before receiving 5-fluorouracil based nCRT. Proteomics analyses were conducted to identify the differentially expressed proteins (DEPs) between total responders (TR) and poor responders (PR). The DEPs were validated via ROC plotter web tool and their predictive performance was evaluated by receiver operating characteristic analysis. Functional enrichment analyses were performed to further explore the potential mechanisms underlying nCRT response. RESULTS: Among 3,998 total proteins, 91 DEPs between TR and PR were screened out. HSPA4, NIPSNAP1, and SPTB all with areas under the curve (AUC) ~ 0.8 in the internal discovery cohort were independently validated by the external mRNA datasets (AUC ~ 0.7), and their protein levels were linearly correlated with the graded responses to nCRT in the internal cohort. The combination of HSPA4 and SPTB could distinctly discriminate the TR and PR groups (AUC = 0.980, p < 0.0001). Moreover, multiple combinations of the three proteins realized increased specificity and/or sensitivity, while achieving favorable predictive value when moderate responders were introduced into the ROC analysis. Pathways including DNA damage repair, cell cycle, and epithelial mesenchymal transition were involved in nCRT response according to the enrichment analysis results. CONCLUSIONS: HSPA4, SPTB and NIPSNAP1 in tumor biopsies and/or their optional combinations might be potential predictive markers for nCRT response in patients with LARC. The DEPs and their related functions have implications for the potential mechanisms of treatment response to nCRT in patients with LARC.
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Puerarin, which is a widely used in Traditional Chinese Medicine, was previously demonstrated to regulate the subsets of CD4+ lymphocytes in gunpowder smog-induced acute lung injury (ALI). However, the underlying mechanism remains largely unknown. Previous studies on autoimmune diseases have revealed that the renin-angiotensin system (RAS) and NF-κB participate in regulating the levels of CD4+ T lymphocytes. The aim of the present study was to further investigate the mechanisms underlying the protective effects of puerarin. Wistar rats were randomly divided into four groups as follows: Normal control, puerarin control, smoke inhalation injury and puerarin treatment plus smoke inhalation injury groups. The levels of angiotensin II (Ang II) in lung tissue and in the circulation, and the levels of interleukin (IL)-6, IL-1ß, IL-17A and tumor necrosis factor (TNF)-α in the bronchoalveolar lavage fluid (BALF) were assayed using ELISA kits. The expression of Ang II type 1 receptor (AT1-R), angiotensin-converting enzyme (ACE) and ACE2 were examined by immunohistochemical analysis and western blotting. Phosphorylated (p-) NF-κB p65 and NF-κB inhibitor α (IκB-α) protein expression levels were also determined using western blotting. Puerarin treatment reduced the levels of inflammatory cytokines in the BALF. Furthermore, puerarin treatment significantly decreased the levels of Ang II, AT1-R and ACE, which were increased following smoke inhalation. Conversely, puerarin treatment upregulated the expression of ACE2, which was downregulated following smoke inhalation. Additionally, puerarin decreased the expression of p-NF-κB p65 and increased that of IkB-α. Thus, the antiinflammatory effects of puerarin were partly mediated via the RAS and via regulation of the NFĸB signaling pathway in rats with gunpowder smog-induced ALI.
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Colorectal cancer (CRC) is the fourth leading cause of cancer deaths worldwide, with poor prognosis mainly related to metastasis. Fibronectin (FN), a vital component of the extracellular matrix (ECM), has been found involved in tumorigenesis and malignant progression in different types of malignancy. Numerous studies have indicated the distinct expression of FN in various cancers and demonstrated the different functions of FN in the proliferation, migration, and invasion of cancers. Meanwhile, FN isoforms have been extensively used for targeted drug delivery and imaging for tumors. Although a growing number of studies on FN in CRC have been reported, integrated reviews on the relationship between FN and CRC are rare. In this review, we will summarize the association between FN and CRC, including the signaling pathways and molecules involved in, as well as potential diagnostic and therapeutic values of FN for patients with CRC.
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Neoplasias Colorrectales/metabolismo , Fibronectinas/química , Transducción de Señal , Empalme Alternativo , Sitios de Unión , Biomarcadores de Tumor/metabolismo , Carcinogénesis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Dimerización , Progresión de la Enfermedad , Disulfuros/química , Sistemas de Liberación de Medicamentos , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Péptidos/química , Pronóstico , Unión Proteica , Isoformas de ProteínasRESUMEN
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.
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Citocromo P-450 CYP2D6/genética , Técnicas de Genotipaje/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Polimorfismo de Nucleótido Simple , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Anciano , Antihipertensivos/sangre , Antihipertensivos/metabolismo , Antihipertensivos/uso terapéutico , Pueblo Asiatico/genética , China , Análisis Costo-Beneficio , Citocromo P-450 CYP2D6/metabolismo , Femenino , Genotipo , Técnicas de Genotipaje/economía , Humanos , Hipertensión/sangre , Hipertensión/tratamiento farmacológico , Hipertensión/genética , Masculino , Metoprolol/sangre , Metoprolol/metabolismo , Metoprolol/uso terapéutico , Persona de Mediana Edad , Reproducibilidad de los ResultadosRESUMEN
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.
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Química Clínica/instrumentación , Química Clínica/métodos , Hematología/instrumentación , Hematología/métodos , Automatización , Automatización de Laboratorios , Recuento de Células Sanguíneas/instrumentación , China , Humanos , Recuento de Leucocitos , Leucocitos/citología , Modelos Lineales , Microscopía , Recuento de Plaquetas , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
An iron scarcity often occurs in chronic kidney disease (CKD). Neutrophil gelatinase-associated lipocalin (NGAL), a biomarker of acute kidney injury, is associated with iron metabolism. The present study determined the association between serum NGAL and iron status in chronic kidney disease with anemia. A total of 154 adult CKD patients were divided into anemia and without anemia groups. The anemia groups were further subdivided into two groups based on the presence or absence of iron deficiency, defined as a transferrin saturation (TSAT) < 20%. The NGAL was measured for all the 154 patients, and the possible relationships with iron status were analyzed. 27.7% patients with TSAT < 20% presented lower hemoglobin, serum iron, serum ferritin, and higher NGAL values than those without iron deficiency. NGAL was inversely correlated with hemoglobin, hematocrit, MCV, MCH, serum iron, and TSAT. NGAL adequately diagnosed the status of iron deficiency among CKD patients by ROC analysis. The optimal NGAL cutoff value able to identify iron deficiency was found to be > 244.8 ng/mL, with 73.01% sensitivity and 68.29% specificity. CKD patients with anemia presented altered NGAL values as this protein is involved in the maintenance of iron balance. Thus, NGAL might be proposed as a new tool for assessing the iron deficiency and in the management of iron therapy for CKD patients.
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Anemia Ferropénica/diagnóstico , Anemia/complicaciones , Hierro/metabolismo , Lipocalina 2/sangre , Insuficiencia Renal Crónica/complicaciones , Regulación hacia Arriba , Adulto , Anciano , Anemia/sangre , Anemia/metabolismo , Anemia Ferropénica/sangre , Biomarcadores/sangre , Comorbilidad , Femenino , Ferritinas/sangre , Humanos , Masculino , Persona de Mediana Edad , Curva ROC , Insuficiencia Renal Crónica/sangre , Insuficiencia Renal Crónica/metabolismo , Sensibilidad y EspecificidadRESUMEN
Quantitative PCR (qPCR) assay using TaqMan™ probe was widely used in the detection of different nucleic acids. However, this technology has several drawbacks, including false negative results caused by primer-dimer (PD) and false positive issues due to primer-probe aggregations. Here, we designed a modified TaqMan™-Molecular Beacon probe by adding an antisense base and a new type of primer pair named central-homo primer pairs bearing 5-10 bases homologous sequence on the 3' end. Using the HBV qPCR assay as a proof of concept, the new design significantly improved the accuracy of the TaqMan™ qPCR assay for HBV detection. Application of the central-homo primer pair led to significantly delayed Ct values by 5-10 cycles compared with conventional primer design. The modified probe containing an antisense base did not produce any detectable signal in repeating primer-probe aggregation experiments. Furthermore, the use of the central-homo primer pair and the non-competitive internal control could solve the false negative problem caused by PD formation. We validated this customized duplex qPCR system using 208 clinical samples collected from patients in clinic showing accuracy was higher than that of the conventional qPCR method.
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Virus de la Hepatitis B/genética , Hepatitis B/diagnóstico , Hepatitis B/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Cartilla de ADN , Femenino , Humanos , Masculino , Sondas Moleculares , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The aim of this study is to evaluate the frequency of CTNNA1 hypermethylation in acute myeloid leukemia (AML) patients in an attempt to improve molecular prognostic model. CTNNA1 promoter methylation levels in 319 newly diagnosed AML patients were detected using quantitative methylation-specific polymerase chain reaction (qMS-PCR). Furthermore, hematological characteristics, cytogenetic abnormalities, and genetic mutation status were analyzed, followed by assessment of clinical impact. Our findings demonstrated that CTNNA1 hypermethylation was observed in 25% AML patients. Hypermethylation of the CTNNA1 promoter was associated with unfavorable karyotype, and also possessed the higher frequency of coexisting with ASXL1 and RUNX1 mutations. Patients with CTNNA1 hypermethylation exhibited the shorter relapse-free survival (RFS) and overall survival (OS) in the whole AML and non-M3 AML patients. Moreover, patients with the higher methylation levels had more aggressive course than those with relative lower levels. In multivariate analyses, CTNNA1 hypermethylation was an independent factor predicting for poor RFS, but not for OS. In conclusion, CTNNA1 hypermethylation may be a reliable factor for improving prognostic molecular model for AML.
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Metilación de ADN , Leucemia Mieloide Aguda/genética , Regiones Promotoras Genéticas/genética , alfa Catenina/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Regulación Leucémica de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Cariotipo , Leucemia Mieloide Aguda/patología , Persona de Mediana Edad , Análisis Multivariante , Mutación , Pronóstico , Proteínas Represoras/genética , Adulto JovenRESUMEN
OBJECTIVE: To explore the value of morphological examination, cytochemical staining combined with bone marrow biopsy in the differential diagnosis between myelodysplastic syndrome (MDS) with low blasts and hemolytic anemia (HA). METHODS: The clinical data of 85 cases of myelodysplastic syndrome with low blasts (< 5%) and 61 patients with hemolytic anemia in Chinese PLA's Gerneral hospital from September 2009 to March 2015 were retrospectively analysed. The clinical characteristics, cytogenetic and molecular features, bone marrow cell count and morphology features, cytochemical staining results and bone marrow biopsy features of above-methioned patients were compared. RESULTS: There was no significant difference (P > 0.05) in clinical data between MDS group and HA group. Megakaryocytic dysplasia-positive rate, and ring sideroblasts positive rate, and PAS positive rate were significantly higher in MDS group than those that in HA group (P < 0.05). Abnormal localization of immature precursors (ALIP) and megakaryocytic dysplasia positive rate in bone marrow biopsy were significantly higher in MDS group than those that in HA group (P < 0.05), 90.6% of MDS with low blasts patients were identifiable by combined detections. CONCLUSION: Combining detection of morphology, cytochemistry staining and bone marrow biopsy has been confirmed to be more useful for differential diagnosis between MDS with low blasts and HA.
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Anemia Hemolítica/diagnóstico , Síndromes Mielodisplásicos/diagnóstico , Anemia Hemolítica/complicaciones , Biopsia , Células de la Médula Ósea/citología , Diagnóstico Diferencial , Células Precursoras Eritroides/citología , Humanos , Megacariocitos/citología , Síndromes Mielodisplásicos/complicaciones , Estudios Retrospectivos , Coloración y EtiquetadoRESUMEN
T helper (Th) 17 cells and CD4(+) CD25(+) regulatory T (Treg) cells are supposed to be critically involved in regulating autoimmune and inflammatory diseases. The aim of this study was to investigate the Th17/Treg pattern in rats with gunpowder smog-induced acute lung injury. Wistar rats were equally randomized to three groups: normal control group, ALI 6 h group (smoke inhalation for 6 h) and ALI 24 h group (smoke inhalation for 24 h). We observed changes in cell counting in bronchoalveolar lavage fluid (BALF), alveolar-capillary membrane permeability and lung tissue pathology. Moreover, rats in ALI 6 h and ALI 24 h group showed increased expression of Th17 cell and related cytokines (IL-17 A, IL-6, TGF-ß and IL-23). Meanwhile, Treg prevalence and related cytokines (IL-10, IL-2 and IL-35) were decreased. Consequently, the ratio of Th17/Treg was higher after smoke inhalation. Additionally, Th1 cell decreased while Th2 cell increased at 6 h and 24 h after smoke inhalation. In conclusion, Th17/Treg imbalance exists in rats with smoke inhalation-induced acute lung injury, suggesting its potential role in the pathogenesis of this disease.
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Lesión Pulmonar Aguda/etiología , Lesión por Inhalación de Humo/complicaciones , Lesión por Inhalación de Humo/inmunología , Linfocitos T Reguladores/inmunología , Células Th17/inmunología , Lesión Pulmonar Aguda/metabolismo , Lesión Pulmonar Aguda/patología , Animales , Líquido del Lavado Bronquioalveolar/citología , Permeabilidad Capilar , Inmunofenotipificación , Recuento de Leucocitos , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Recuento de Linfocitos , Subgrupos Linfocitarios/inmunología , Subgrupos Linfocitarios/metabolismo , Peroxidasa/metabolismo , Fenotipo , Ratas , Humo/efectos adversos , Humo/análisis , Lesión por Inhalación de Humo/patología , Linfocitos T Reguladores/metabolismo , Células Th17/metabolismoRESUMEN
BACKGROUND: The diagnosis of myelodysplastic syndrome (MDS), especially hypoplastic MDS, and MDS with low blast counts or normal karyotype may be problematic. This study characterized ID4 gene methylation in patients with MDS and aplastic anemia (AA). METHODS: The methylation status of ID4 was analyzed by bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time methylation-specific PCR (MethyLight PCR) in 100 patients with MDS and 31 patients with AA. RESULTS: The MDS group had a higher ID4 gene methylation positivity rate (22.22%) and higher methylation levels (0.21 [0-3.79]) than the AA group (P < 0.05). Furthermore, there were significant differences between the hypoplastic MDS and AA groups, the MDS with low blast count and the AA groups, and the MDS with normal karyotype and the AA groups. The combination of genetic and epigenetic markers was used in much more patients with MDS (62.5% [35/56]) than the use of genetic markers only (51.79% [29/56]). CONCLUSIONS: These results showed that the detection of ID4 methylation positivity rates and levels could be a useful biomarker for MDS diagnosis.
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Anemia Aplásica/genética , Proteínas Inhibidoras de la Diferenciación/genética , Síndromes Mielodisplásicos/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Islas de CpG/genética , Metilación de ADN/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
OBJECTIVE: Puerarin, a kind of traditional Chinese medicine, possesses immunomodulatory property. However, the immunomodulatory effects of puerarin on smoke inhalation injury have not been determined. The aim of the current study was to investigate the therapeutic efficacy of puerarin on gunpowder smog-induced acute lung injury in rats via regulation of Th1/Th2/Th17 expression. MATERIALS AND METHODS: Wistar rats were equally randomized to four groups (normal control group, puerarin control group, smoke inhalation injury group, puerarin treatment plus smoke inhalation injury group). The severity of lung injury was evaluated by histopathology, myeloperoxidase (MPO) activity in lung homogenates, cell counting in bronchoalveolar lavage fluid (BALF), and lung vascular permeability parameters including lung wet to dry weight ratio and protein concentration in BALF. Flow cytometry was used to analyze the expression of Th1/Th2/Th17 lymphocytes in blood of rats. RESULTS: Puerarin showed significant therapeutic effects against neutrophil infiltration and tissue injury, as evidenced by histopathological findings and MPO activity. Lung vascular permeability was also relieved by puerarin administration. Additionally, puerarin significantly decreased the number of neutrophils and lymphocytes in BALF compared with smoke inhalation injury group. Furthermore, puerarin increased Th1 immunity and reduced Th2 and Th17 responses and thereby altering the Th1/Th2/Th17 imbalance induced by smoke inhalation. CONCLUSIONS: Our findings suggested that puerarin suppressed inflammatory responses in gunpowder smog-induced acute lung injury by regulation of Th1/Th2/Th17 expression, and may be a potential therapeutic agent for smoke inhalation injury.
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Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Isoflavonas/farmacología , Isoflavonas/uso terapéutico , Lesión por Inhalación de Humo/tratamiento farmacológico , Lesión por Inhalación de Humo/inmunología , Animales , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Permeabilidad Capilar/efectos de los fármacos , Recuento de Células , Citocinas/inmunología , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Pulmón/patología , Masculino , Peroxidasa/metabolismo , Ratas Wistar , Lesión por Inhalación de Humo/patología , Células TH1/inmunología , Células Th17/inmunología , Células Th2/inmunologíaRESUMEN
OBJECTIVE: To investigate the clinical significance of ZO-1 gene methylation level in MDS progression in order to provide a theoretical basis for evaluating progrosis of MDS patients. METHODS: The methylation specific PCR (MS-PCR) was performed to evaluate the ZO-1 gene methylation status in bone marrow samples of normal persons as control (NC). MDS and AML patients, the bisulfite sequencing PCR (BSP) was applied to detect the ZO-1 gene methylation status in serial bone marrow samples of MDS-RA, MDS-RAEB and AML stages of a MDS patients. RESULTS: The possitive rate of ZO-1 gene methylation in samples of NC, MDS and AML patients displayed significant difference; in sample of NC group the positive of ZO-1 gene methylation was not observed, but the positive rate of ZO-1 gene methylation in samples of AML patients was highest (65.0%), the proportion of ZO-1 gene methylation in myeloid blast count of MDS/AML patients was higher (P=0.000). The serial samples in one MDS patient showed that along with progress of disease, the positive rate of ZO-1 gene methylation in MDS-RA, MDS-RAEB and AML stages was found to be obvious different (P=0.000), the positive rate of ZO-1 gene methylation in AML stage was highest (64.65%). CONCLUSION: The high methylation in promoter region of ZO-1 gene has been found in MDS/AML patients, and along with clonal proliferation, the positive rate of ZO-1 methylation and positive froguency of methylation sites increase graduatly which suggests that the MDS progresses in a certain degree, and the ZO-1 gene methylation level may be used as an new indicator for monitoring desease progression from MDS to AML.
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Metilación de ADN , Síndromes Mielodisplásicos , Médula Ósea , Progresión de la Enfermedad , Humanos , Leucemia Mieloide Aguda , Metilación , Reacción en Cadena de la Polimerasa , Proteína de la Zonula Occludens-1RESUMEN
OBJECTIVE: This study was aimed to evaluate the significance of bone marrow(BM) morphological examination and many tumor marker(TM) detection, especially carcinoembryonic antigen (CEA), cancer antigen 125(CA125), cancer antigen 15-3 (CA15-3) and serum ferritin (SF) for lymphoma diagnosis and prognosis. METHODS: A total of 47 confirmed patients with lymphoma in our hospital from January 2012 to October 2013 and 20 health peoplels as normal controls were performed with bone marrow morphological examination, at the same time, the electrochemistry luminescent technique was applied for detecting levels of TM (especially CEA, CA125, CA15-3 and SF) in serum samples of lymphoma patient and normal controls, then the BM immature lymphocyte counts of these people and clinical parameters were analyzed for diagnosis and prognosis. RESULTS: There was significant differences in all the four TM levels between serum samples of lymphoma patients and normal control (P=0.029, P=0.000, P=0.005, P=0.000). These TM levels had no correlation with age, sex white blood cell, lymphocyte, platelet counts and anemia of lymphoma patients (P>0.05). It was also found that the patients with elevated TM levels had high BM immature lymphocytes (lymphoma cells) counts, B symptoms, advanced clinical stage and high IPI index (P<0.05). The CA15-3 and SF levels in serum samples of lymphoma patients with BM infiltration were higher than that in lymphoma patients without BM infiltration (P=0.002, P=0.000). CONCLUSION: Combination of BM morphological examination with serum TM level detection plays an important role in diagnosis, clinical stage and prognosis evaluation of lymphoma patients. It is also very important for assessing BM infiltration status of lymphoma patients.
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Médula Ósea , Biomarcadores de Tumor , Examen de la Médula Ósea , Antígeno Ca-125 , Antígeno Carcinoembrionario , Humanos , Linfoma , PronósticoRESUMEN
OBJECTIVE: To evaluate significance of ID4 gene mehtylation in demethylating myelodysplastic syndrome(MDS) cell Line MUTZ1 and 2 patients with MDS. METHODS: The methylation-specific PCR (MS-PCR) and reverse transcription-PCR (RT-PCR) were applied to identify the methylation status and gene expression of ID4 gene in MDS cell line MUTZ1, a patient with aplastic anemia(AA) and a donor with normal bone marrow (NBM). RT-PCR was applied to detect the ID4 gene expression status in MUTZ1 cell line treated with decitabine at 3 different concentrations. Then bisulfite sequencing PCR (BSP) was applied to detect ID4 gene methylation status in 2 MDS parients treated with decitabine. RESULTS: The MDS cell line MUTZ-1 displayed a complete methylation of ID4 gene promoter with little mRNA expression. Inversely, bone marrow of an AA patient and NBM showed complete unmethylation of this gene with intensity mRNA expression. With the increase of decitabine concentration, ID4 gene mRNA expression was more and more increased. After decitabine treatment, ID4 gene methylation-positive frequencies of both the 2 MDS patients were much more decreased than that of the first treatment. So, ID4 gene mRNA expression inhibited by promoter hypemethylation could be recovered by using demethylation medicine. CONCLUSION: ID4 as a new potential anti-oncogene suggests that its methylation may become a marker for selection and assessment of therapeutic schedules in patients with MDS.
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Metilación de ADN , Síndromes Mielodisplásicos , Anemia Aplásica , Azacitidina/análogos & derivados , Médula Ósea , Línea Celular , Decitabina , Expresión Génica , Genes Supresores de Tumor , Humanos , Proteínas Inhibidoras de la Diferenciación , Reacción en Cadena de la Polimerasa , Regiones Promotoras GenéticasRESUMEN
BACKGROUND: Myelodysplastic syndrome (MDS) eventually transforms into acute leukemia (AL) in about 30% of patients. Hypermethylation of the inhibitor of DNA binding 4 (ID4) gene may play an important role in the initiation and development of MDS and AL. The aim of this study was to quantitatively assess ID4 gene methylation in MDS and to establish if it could be an effective method of evaluating MDS disease progression. METHODS: We examined 142 bone marrow samples from MDS patients, healthy donors and MDS-AL patients using bisulfite sequencing PCR and quantitative real-time methylation-specific PCR. The ID4 methylation rates and levels were assessed. RESULTS: ID4 methylation occurred in 27 patients (27/100). ID4 gene methylation was more frequent and at higher levels in patients with advanced disease stages and in high-risk subgroups according to WHO (P < 0.001, P < 0.001, respectively) and International Prognostic Scoring System (IPSS) (P = 0.002, P = 0.007, respectively) classifications. ID4 methylation levels changed during disease progression. Both methylation rates and methylation levels were significantly different between healthy donor, MDS patients and patients with MDS-AL (P < 0.001, P < 0.001, respectively). Multivariate analysis indicated that the level of ID4 methylation was an independent factor influencing overall survival. Patients with MDS showed decreased survival time with increased ID4 methylation levels (P = 0.011, hazard ratio (HR) = 2.371). Patients with ID4 methylation had shorter survival time than those without ID4 methylation (P = 0.008). CONCLUSIONS: Our findings suggest that ID4 gene methylation might be a new biomarker for MDS monitoring and the detection of minimal residual disease.
Asunto(s)
Metilación de ADN , Proteínas Inhibidoras de la Diferenciación/genética , Síndromes Mielodisplásicos/genética , Adulto , Estudios de Casos y Controles , Humanos , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/diagnósticoRESUMEN
BACKGROUND: A revised classification of chronic kidney disease (CKD) was proposed by the Kidney Disease: Improving Global Outcomes (KDIGO) in 2012. Neutrophil gelatinase-associated lipocalin (NGAL) was considered as one of the most promising biomarkers in clinical nephrology. The aim of this study was to examine the level of NGAL in patients with different impairment of GFR based on the new classification, and to evaluate whether NGAL in serum or urine was associated with different risk categories in CKD patients. METHODS: A cross-sectional study was performed in 240 patients with CKD. NGAL, serum cystatin C, ß2-macroglobulin (ß2-MG), urine α1-macroglobulin (α1-MG) and albuminuria were tested in patients with various degrees of renal impairment. RESULTS: Good correlation was found between the NGAL and the cystatin C, ß2-MG and the α1-MG (r > 0.7). The level of sNGAL in CKD stage 3b was more than that in CKD stage 3a (P = 0.025). The concentration of the NGAL increased progressively with the increasing of risk categories (proposed by the revised CKD classification). The cutoff value of NGAL was calculated from stage 2 to stage 5. ROC analysis showed good AUC (sNGAL > 0.8, uNGAL > 0.7) and high specificity (sNGAL > 87%, uNGAL > 90%) on the cutoff value of NGAL. CONCLUSION: The results confirm NGAL as a useful biomarker in clinical nephrology which is helpful to diagnosis and evaluate the categories for CKD proposed by the KDIGO.