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BACKGROUND: There has been much research into the impact of shift systems on clinicians and nurses, but little research into quality control in clinical laboratories. This topic focuses on assessing the impact of shift systems on clinical laboratory scientists. METHODS: A total of 34,955 CBCs from pediatric patients who visited the hospital during night-time hours over a period of three years were selected for analysis. The quality of routine blood tests was evaluated using four indica-tors: red blood cell count, white blood cell count, platelet count, and hemoglobin levels. The effects of gender, years of experience, and length of the night shift on test results were evaluated separately for each clinical laboratory scientist. RESULTS: The results showed that the gender and years of experience of the clinical laboratory scientists did not affect the CBC results. However, a significant impact was observed as the number of hours worked on night shifts increased. CONCLUSIONS: The findings of this study suggest that the night shift schedule of clinical laboratory scientists can have an impact on the accuracy of pediatric CBCs. It is essential for healthcare institutions to consider the length of night shifts for clinical laboratory scientists and implement measures to minimize the impact on test results.
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Servicios de Laboratorio Clínico , Horario de Trabajo por Turnos , Humanos , Niño , Laboratorios Clínicos , Recuento de Células Sanguíneas , Recuento de PlaquetasRESUMEN
We report a case of hemophagocytic syndrome (HPS) secondary to brucellosis, in which typhoidal cells were found in bone marrow, suggesting typhoidal cells present not only in Salmonella typhi infections but also in other bacterial infections. Typhoidal cells in bone marrow can be used to quickly identify the presence of bacterial infection pending the results of bone marrow and/or blood cultures.
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Brucelosis , Linfohistiocitosis Hemofagocítica , Fiebre Tifoidea , Femenino , Humanos , Fiebre Tifoidea/complicaciones , Fiebre Tifoidea/microbiología , Linfohistiocitosis Hemofagocítica/etiología , Brucelosis/complicacionesRESUMEN
BACKGROUND: The COVID-19 outbreak, which began in late 2019, continues to ravage the globe and has become the greatest threat to human health. As nucleic acid test is the primary means of screening for COVID-19, this makes the laboratory the most important node in the epidemic prevention and control system. METHODS: As a small laboratory in the hospital, we can meet a large number of demands for nucleic acid test by optimizing staff process, strictly disinfecting experimental batches and changing experimental methods. RESULTS: Through the improvement of the above aspects, our daily maximum detection quantity has been increased from 256/day to 1,012/day. Besides, none of the medical staff has been infected. And there have been no nosocomial infections. CONCLUSIONS: Nucleic acid laboratories, especially small laboratories, should promptly adjust their strategies in the face of unexpected outbreaks and conduct risk assessment in accordance with laboratory activities.
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Prueba de Ácido Nucleico para COVID-19 , COVID-19/diagnóstico , COVID-19/virología , Necesidades y Demandas de Servicios de Salud/organización & administración , Tamizaje Masivo/organización & administración , Manejo de Especímenes , Flujo de Trabajo , Carga de Trabajo , Humanos , Control de Infecciones/organización & administración , Salud Laboral , Valor Predictivo de las PruebasRESUMEN
Preeclampsia (PE) is a specific obstetric disorder that may result in maternal and neonatal morbidity and mortality. Increasing evidence has been indicated that some candidate genes related to oxidative stress, such as glutamate-cysteine ligase, catalytic subunit (GCLC), glutamate-cysteine ligase, modifier subunit (GCLM), involve in the pathogenesis of PE. After the genetic contribution of GCLC rs17883901 polymorphism was analyzed by TaqMan allelic discrimination real-time PCR in 1001 PE patients and 1182 normal pregnant women, a case-control association analysis was performed. Although no statistical difference was found in genetic distribution of rs17883901 in GCLC between PE and control group (χ2 = 2.201, p = .333 by genotypic, χ2 = 0.524, p = .469, OR = 0.932, 95%CI = 0.771-1.128 by allelic), significant differences in the genotypic frequencies were investigated between mild PE group (χ2 = 6.999, p = .030) or late-onset PE group (χ2 = 6.197, p = .045) and control group. Furthermore, when dividing the mild PE patients, the late-onset PE patients and the controls into TT/CT + CC, TT + CT/CC, and TT/CC subgroups, we found statistical differences between mild PE and controls (TT/CT + CC:χ2 = 5.132, p = .023, OR = 2.948, 95%CI = 1.107-7.854; TT/CC:χ2 = 4.564, p = .033, OR = 2.793, 95%CI = 1.046-7.460) as well as late-onset PE and controls (TT/CT + CC:χ2 = 4.043, p = .044, OR = 2.248, 95%CI = 1.000-5.055). This is the first study to indicate GCLC rs17883901 polymorphism may be associated with a risk of mild PE and late-onset PE in Chinese Han women. However, additional well-designed studies with multi-ethnic and large-scale samples should be performed to validate our results.
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Glutamato-Cisteína Ligasa/genética , Polimorfismo de Nucleótido Simple , Preeclampsia/genética , Adulto , Pueblo Asiatico/genética , Pueblo Asiatico/estadística & datos numéricos , Estudios de Casos y Controles , China/epidemiología , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Preeclampsia/etnología , Embarazo , Adulto JovenRESUMEN
Objectives: The involvement of oxidative stress in the pathophysiology of preeclampsia (PE) has been already suggested. In this present study, we aimed to investigate the association of the genetic frequency of heme oxygense-1 (HMOX1) polymorphism with PE in Chinese Han women.Methods: We researched the genetic distribution of rs2071746 polymorphism in HMOX1 by the TaqMan allelic discrimination real-time PCR between 1235 PE patients and 1720 healthy women.Results: We found there were't significant differences in the distribution of HMOX1 rs2071746 polymorphism in PE compared to the control group (rs2071746, genotype χ2 = 0.282, P = 0.869 and allele χ2 = 0.027, P = 0.869, OR = 1.009, 95% = 0.909-1.120).Conclusion: The rs2071746 polymorphism in HMOX1 might not be related to PE in Chinese women, although further investigations should be conducted to confirm our findings.
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Hemo-Oxigenasa 1/genética , Preeclampsia/genética , Adulto , Alelos , Pueblo Asiatico/genética , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad/genética , Genotipo , Humanos , Hipertensión/genética , Polimorfismo de Nucleótido Simple , Embarazo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Previous studies have indicated that an increased inflammatory response plays an important role in preeclampsia (PE), and rising levels of interleukin (IL)-22 can trigger inflammation and hyperproliferation, leading to increased production of several pro-inflammatory cytokines such as IL-1, IL-6, and IL-8. We aimed to investigate the association between polymorphisms of IL-22 and IL-22 receptor alpha 1 gene (IL-22RA1) and PE in Chinese Han population. Single nucleotide polymorphisms (SNPs) rs2227485 in IL-22 and rs3795299 in IL-22RA were genotyped by Taqman real-time PCR in 1071 PE patients and 1263 control subjects. Differences in genetic distribution were compared between two groups using the chi-square test. Significant differences were observed in genotypic and allelic frequencies of IL-22RA1 rs3795299 between healthy controls and PE patients (P < 0.001 by genotype; P = 0.001, odds ratio = 1.253, 95% confidence interval 1.103-1.424 by allele). There were also significant differences in genotypic and allelic frequencies of rs3795299 between late-onset/mild PE and control groups. In addition, we found obvious statistic difference for the allele of early-onset PE/the genotype of late-onset PE and control subgroups for IL-22 rs2227485. IL-22 rs2227485 and IL-22RA1 rs3795299 may be associated with the development of PE in Chinese Han population. However, further validation is required in other populations, as well as an evaluation of the association of other SNPs in IL-22 and IL-22RA1 with PE.
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Alelos , Frecuencia de los Genes , Interleucinas/genética , Polimorfismo Genético , Preeclampsia/genética , Receptores de Interleucina/genética , Adulto , Pueblo Asiatico , China/epidemiología , Femenino , Humanos , Preeclampsia/epidemiología , Embarazo , Factores de Riesgo , Interleucina-22RESUMEN
BACKGROUND/AIMS: Preeclampsia (PE) is a systemic inflammatory response syndrome involving varieties of cytokines, and previous studies have shown that IL-33 and its receptor IL-1RL1 play pivotal roles in the development of it. As a polygenetic hereditary disease, it is necessary to study the gene analysis for PE. Therefore, the present study was to determine whether IL-33 rs3939286 and IL-1RL1 rs13015714 associated with susceptibility to PE in Chinese Han women. METHODS: 1,031 PE patients and 1,298 controls were enrolled and the genotyping for rs3939286 in IL-33 and rs13015714 in IL-1RL1 was performed by TaqMan allelic discrimination real-time PCR. Hardy-Weinberg equilibrium (HWE) was examined to ensure the group representativeness and Pearson's chi-square test was used to compare the differences in genetic distributions between the two groups. RESULTS: No significant differences in genotypic and allelic frequencies of the two polymorphisms loci were observed between cases and controls. There were also no significant differences in genetic distributions between mild/severe and early/late-onset PE and control groups. CONCLUSION: Although our data suggested that the polymorphisms of IL-33 rs3939286 and IL-1RL1 rs13015714 might not be critical risk factors for PE in Chinese Han women, the results need to be validated in different nations.
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Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-33/genética , Preeclampsia/genética , Alelos , Pueblo Asiatico/genética , Estudios de Casos y Controles , China , Femenino , Frecuencia de los Genes , Humanos , Oportunidad Relativa , Polimorfismo de Nucleótido Simple , Embarazo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
BACKGROUND/AIMS: Several lines of evidence have been reported that oxidative stress plays an important role in the pathogenesis of Preeclampsia (PE). Therefore, this research is aimed to investigate whether polymorphisms of CYBA are related to susceptibility to PE in Chinese Han women. METHODS: We studied the genetic frequency of the rs9932581 and 1049255 polymorphisms in CYBA in 1029 PE patients and 1400 controls of later pregnant women by the TaqMan allelic discrimination real-time PCR and a case-control model. RESULTS: Our research indicated that no significant differences were found for the genotypic or allelic frequencies at the two polymorphic sites in CYBA between PE patients and controls. To further study the relationship between the polymorphic sites and PE, we also found that there is no significant difference in the genetic distributions identified between the mild or severe PE and early or the late-onset PE and controls. CONCLUSION: The study demonstrated that the genetic variants of rs9932581 and rs1049255 in CYBA might not be associated with PE. However, investigations of genetic variability that influence on the disease outcome are needed in other large prospective populations or regions, so the complicated interconnection of genetic and environmental elements can be emulated for better understanding.
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NADPH Oxidasas/genética , Polimorfismo de Nucleótido Simple , Preeclampsia/genética , Adulto , Alelos , Pueblo Asiatico , Estudios de Casos y Controles , Femenino , Expresión Génica , Frecuencia de los Genes , Genotipo , Humanos , Preeclampsia/diagnóstico , Preeclampsia/etnología , EmbarazoRESUMEN
IL-33 is a member of IL-1 superfamily that drives production of Th2-related cytokines. Recently, accumulating evidence suggest an involvement of IL-33 in carcinogenesis. Herein, we determine a close correlation of IL-33 expression and cancer progress in patients with non-small-cell lung cancer (NSCLC). Overexpression of IL-33 by transfection with IL-33 expression vector enhances NSCLC outgrowth and metastasis, while genetic knockdown of IL-33 by transfection with IL-33 shRNA limits NSCLC progression. In consistent, IL-33 stimulation of NSCLC cells leads to robust NSCLC outgrowth and metastasis in vitro and in vivo. Mechanically, IL-33-triggered NSCLC progression relies on ST2 receptor and could be abrogated by ST2 blockade. IL-33/ST2 pathway up-regulates membrane glucose transporter 1 (GLUT1) on NSCLC cells, enhancing their glucose uptake and glycolysis. Accordingly, interfering GLUT1 expression dampens IL-33-enhanced glucose uptake and glycolysis in NSCLC cells, thereby abrogates IL-33-induced NSCLC outgrowth and metastasis. In essence, these findings derived from patients' NSCLC cells uncover a new function of IL-33 in NSCLC pathogenesis and identify GLUT1 as a novel target of IL-33 signaling. Block IL-33 is a promising therapeutic strategy to limit NSCLC glycolysis and tumor progression in clinical practice.
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Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Interleucina-33/metabolismo , Neoplasias Pulmonares/metabolismo , Metástasis de la Neoplasia , Animales , Proliferación Celular , Progresión de la Enfermedad , Femenino , Citometría de Flujo , Glucosa/química , Transportador de Glucosa de Tipo 1/metabolismo , Glucólisis , Humanos , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Ratones , Ratones Endogámicos BALB C , Invasividad Neoplásica , Transducción de SeñalRESUMEN
OBJECTIVE: To investigate whether IL-12B rs3212227 was associated with susceptibility to PE (preeclampsia) in Chinese Han women. METHODS: We enrolled 1000 PE patients and 1287 controls and performed rs3212227 genotyping by PCR-restriction fragment length polymorphism. RESULTS: No significant differences in genetic distributions of IL-12B rs3212227 were observed between cases and controls (χ(2) = 4.62, p = 0.10 by genotype; χ(2) = 0.03, p = 0.87 by allele). There were also no significant differences in genotypic and allelic frequencies between mild/severe or early/late-onset PE and control subgroups. CONCLUSION: Our data suggested that IL-12B rs3212227 might not be a critical risk factor for PE in Chinese Han women.
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Subunidad p40 de la Interleucina-12/genética , Preeclampsia , Adulto , Alelos , China/epidemiología , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Polimorfismo de Nucleótido Simple , Preeclampsia/epidemiología , Preeclampsia/genética , EmbarazoRESUMEN
OBJECTIVES: We aimed to explore the relationship between clinical characteristics and circulating lymphocyte profiles in Chinese male patients with primary Sjögren's syndrome (pSS). METHOD: Data from 397 patients with pSS were analyzed retrospectively. 37 were male, which is a prevalence of 9.3%. The clinical, laboratory, and immunophenotypic profiles of peripheral blood lymphocyte subsets were compared between male and female pSS patients. RESULTS: Male patients with primary Sjögren's syndrome have unique clinical manifestations and circulating lymphocyte profiles. Male patients complained more about xerophthalmia and presented with more extra-glandular manifestations as compared with female patients. The CD4+/CD8+ ratio (P = 0.030), the prevalence of CD4-CD8- T cells in lymphocytes (P = 0.020), the absolute number of CD4-CD8- T cells (P = 0.035), the prevalence of CD4+ T cells in lymphocytes (P < 0.001), and the absolute number of CD4+ T cells (P = 0.023) were significantly lower in male patients compared to female patients. On the other hand, the prevalence of CD8+CD28+ T cells (P = 0.030) and CD4+CD25high T cells (P = 0.040) in lymphocytes was significantly higher in male patients than in female patients. Moreover, compared to females with pSS, an elevated serum IgG level, low C3 and C4 levels, anti-SSB positivity, and ANA titers of ≥ 1:160 positivity were more frequent in male with pSS. CONCLUSIONS: Male patients with pSS have distinctive peripheral blood lymphocyte subpopulations, present with more severe clinical symptoms and immunological features, and have an unfavorable prognosis. Key Points ⢠Male patients with pSS have more severe clinical symptoms and specific characteristics of peripheral blood lymphocyte subsets. ⢠Male pSS patients exhibit a higher intensity of the disease (as evaluated by ESSDAI). ⢠Male patients with pSS require individualized treatment regimens and closer follow-up.
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Síndrome de Sjögren , Humanos , Síndrome de Sjögren/inmunología , Síndrome de Sjögren/sangre , Síndrome de Sjögren/complicaciones , Masculino , Persona de Mediana Edad , Femenino , Estudios Retrospectivos , Adulto , Factores Sexuales , Anciano , Relación CD4-CD8 , Subgrupos Linfocitarios/inmunologíaRESUMEN
Recent data support the role of S100A10 in tumorigenesis. In this study, we evaluated the value of S100A10 positivity as a possible biomarker in colorectal cancer. We evaluated S100A10 positivity by immunohistochemistry in a large population of colorectal cancer patients (n = 882). The relationships between S100A10 positivity and clinicopathological features and clinical outcome were analyzed. There were 36 % (319/882) tumors positive for S100A10 in all colorectal cancer samples. In contrast, normal colorectal epithelium was negative for S100A10 among all 562 specimens of adjacent normal mucosa. S100A10 positivity was correlated with poor differentiation (p = 0.0012) and disease stage (p = 0.003). S100A10 positivity was significantly correlated with shortened specific [log-rank p < 0.001; multivariate hazard ratio (HR), 1.49; 95% confidence interval (CI), 1.09-2.04] and overall survival (log-rank p = 0.0012; multivariate HR, 1.34; 95% CI, 1.06-1.73). Knockdown of S100A10 by siRNA significantly reduced the proliferation, migration, and invasion capacity of colorectal cancer cell lines. Our results suggest a role for S100A10 as a prognostic marker and potential therapeutic target in colorectal cancer.
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Anexina A2/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias Colorrectales/metabolismo , Proteínas S100/metabolismo , Anciano , Anexina A2/genética , Western Blotting , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Femenino , Células HCT116 , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Análisis Multivariante , Invasividad Neoplásica , Pronóstico , Interferencia de ARN , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas S100/genéticaRESUMEN
Purpose: Klebsiella pneumoniae, a gram-negative bacterium, poses a severe hazard to public health, with many bacterial hosts having developed resistance to most antibiotics in clinical use. The goal of this study was to look into the development of resistance to both ceftazidime-avibactam and carbapenems, including imipenem and meropenem, in a K. pneumonia strain expressing a novel K. pneumoniae carbapenemase-2 (KPC-2) variant, referred to as KPC-49. Methods: After 1 day of incubation of K1 on agar containing ceftazidime-avibactam (MIC = 16/4 mg/L), a second KPC-producing K. pneumoniae strain (K2) was recovered. Antimicrobial susceptibility assays, cloning assays, and whole genome sequencing were performed to analyse and evaluate antibiotic resistance phenotypes and genotypes. Results: K. pneumoniae strain (K1), that produced KPC-2, was susceptible to ceftazidime-avibactam but resistant to carbapenems. The K2 isolate harboured a novel bla KPC-49 variant, which differs from bla KPC-2 by a single nucleotide (C487A), and results in an arginine-serine substitution at amino acid position 163 (R163S). The mutant K2 strain was resistant to both ceftazidime-avibactam and carbapenems. We demonstrated the ability of KPC-49 to hydrolyse carbapenems, which may be attributed to high KPC-49 expression or presence of an efflux pump and/or absence of membrane pore proteins in K2. Furthermore, blaKPC-like was carried on an IncFII (pHN7A8)/IncR-type plasmid within a TnAs1-orf-orf-orf-orf-orf-orf-ISKpn6-bla KPC-ISKpn27 structure. The bla KPC-like gene was flanked by various insertion sequences and transposon elements, including the Tn3 family transposon, such as TnAs1, TnAs3, IS26, and IS481-ISKpn27. Conclusion: New KPC variants are emerging owing to sustained exposure to antimicrobials and modifications in their amino acid sequences. We demonstrated the drug resistance mechanisms of the new mutant strains through experimental whole genome sequencing combined with bioinformatics analysis. Enhanced understanding of laboratory and clinical features of infections due to K. pneumoniae of the new KPC subtype is key to early and accurate anti-infective therapy.
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In order to improve the resource utilization of recycled concrete powder (RCP), this study aimed to investigate the effect of RCP admixture, curing age, and alkali excitation on the strength of RCP concrete. In addition, the pore structure characteristics of RCP concrete were analyzed in combination with low-field NMR. Furthermore, a gray predictive GM (1, 4) model was established to predict the mechanical properties of the concrete based on the pore structure parameters, especially the compressive and flexural tensile strengths. The results of the study indicate that the mechanical properties, namely compressive strength and flexural strength, of RCP concrete exhibit an initial increase followed by a subsequent decrease with increasing RCP content at 3 d, 7 d, and 28 d curing ages. In particular, the concrete exhibits the highest mechanical properties when the RCP content reaches 10%. As the curing age increases, the RCP gradually achieves full hydration, resulting in further refinement of the concrete pores and a denser structure, which subsequently improves the mechanical properties. In addition, the strength growth rate of alkali-excited recycled concrete (ARC) showed a continuous increase, indicating that alkali excitation increasingly improved the mechanical properties of the concrete. Furthermore, the study accurately predicted the mechanical properties of RCP concrete by using GM (1, 4) prediction models for its compressive strength and flexural tensile strength using pore characteristic parameters.
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Being a key industrial enzyme, tannase is extensively applied in various fields. Despite the characterizations of a large number of tannases, there are hardly a few tannases with exceptional thermostability. In this detailed study, a tannase-encoding gene named tanA was identified from Aureobasidium melanogenum T9 and heterologously expressed in Yarrowia lipolytica host of food grade. The purified tannase TanA with a molecular weight of above 63.0 kDa displayed a specific activity of 941.4 U/mg. Moreover, TanA showed optimum activity at 60°C and pH 6.0. Interestingly, TanA exhibited up to 61.3% activity after incubation for 12 h at 55°C, signifying its thermophilic property and distinguished thermostability. Additionally, TanA was a multifunctional tannase with high specific activities to catalyze the degradation of various gallic acid esters. Therefore, this study presents a novel tannase, TanA, with remarkable properties, posing as a potential candidate for food and agricultural processing.
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Clinically, red blood cell abnormalities are closely related to tumor diseases, red blood cell diseases, internal medicine, and other diseases. Red blood cell classification is the key to detecting red blood cell abnormalities. Traditional red blood cell classification is done manually by doctors, which requires a lot of manpower produces subjective results. This paper proposes an Attention-based Residual Feature Pyramid Network (ARFPN) to classify 14 types of red blood cells to assist the diagnosis of related diseases. The model performs classification directly on the entire red blood cell image. Meanwhile, a spatial attention mechanism and channel attention mechanism are combined with residual units to improve the expression of category-related features and achieve accurate extraction of features. Besides, the RoI align method is used to reduce the loss of spatial symmetry and improve classification accuracy. Five hundred and eighty eight red blood cell images are used to train and verify the effectiveness of the proposed method. The Channel Attention Residual Feature Pyramid Network (C-ARFPN) model achieves an mAP of 86%; the Channel and Spatial Attention Residual Feature Pyramid Network (CS-ARFPN) model achieves an mAP of 86.9%. The experimental results indicate that our method can classify more red blood cell types and better adapt to the needs of doctors, thus reducing the doctor's time and improving the diagnosis efficiency.
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The classification of six types of white blood cells (WBCs) is considered essential for leukemia diagnosis, while the classification is labor-intensive and strict with the clinical experience. To relieve the complicated process with an efficient and automatic method, we propose the Attention-aware Residual Network based Manifold Learning model (ARML) to classify WBCs. The proposed ARML model leverages the adaptive attention-aware residual learning to exploit the category-relevant image-level features and strengthen the first-order feature representation ability. To learn more discriminatory information than the first-order ones, the second-order features are characterized. Afterwards, ARML encodes both the first- and second-order features with Gaussian embedding into the Riemannian manifold to learn the underlying non-linear structure of the features for classification. ARML can be trained in an end-to-end fashion, and the learnable parameters are iteratively optimized. 10800 WBCs images (1800 images for each type) is collected, 9000 images and five-fold cross-validation are used for training and validation of the model, while additional 1800 images for testing. The results show that ARML achieving average classification accuracy of 0.953 outperforms other state-of-the-art methods with fewer trainable parameters. In the ablation study, ARML achieves improved accuracy against its three variants: without manifold learning (AR), without attention-aware learning (RML), and AR without attention-aware learning. The t-SNE results illustrate that ARML has learned more distinguishable features than the comparison methods, which benefits the WBCs classification. ARML provides a clinically feasible WBCs classification solution for leukemia diagnose with an efficient manner.
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Atención , LeucocitosRESUMEN
OBJECTIVE: To observe the effect of signal transducers and activators of transcription 3 (STAT3) gene silence on the growth of breast cancer cell line MCF7 in vitro and in vivo and discuss the feasibility and effectiveness of STAT3 used as gene therapeutic target for breast cancer. METHODS: Human breast cancer cell line MCF7 cells were divided into 3 groups: mock control group, control group transfected with scrambled sequence siRNA, and experimental group transfectod with STAT3 siRNA. The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were examined by MTT method and flow cytometry. MCF7 cells treated with STAT3-siRNA were transplanted subcutaneously in nude mice and their tumorgenic ability was observed. The STAT3 mRNA and protein levels of the samples from nude mice of different groups were detected by semi-quantity RT-PCR and Western blotting and compared. RESULTS: After treatment with STAT3-siRNA, STAT3 mRNA (0.327 ± 0.020 vs. 1.035 ± 0.050, 1.093 ± 0.018) and ptotein (0.153 ± 0.006 vs. 1.320 ± 0.033, 1.374 ± 0.022) levels in the MCF7 cells transfected with STAT3-siRNA were significantly lower than that in the two control groups (P < 0.05). MTT assay showed that after transfection of the STAT3-siRNA into MCF7 cells, cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was (44.00 ± 5.10)%, significantly higher than that in the control group (16.1 ± 1.05)% (P < 0.05). Flow cytometry results suggested that more apoptosis was observed in the STAT3-siRNA group. The apoptosis rate was (14.79 ± 0.22)%, much higher than that in the control group [(7.06 ± 0.71)%, (8.45 ± 0.43)%, P < 0.05]. The tumor growth in the experimental group was significantly slower than that in the two control groups. 0n the 22th day after transplantation, the tumor weight [(21.4 ± 10.6) mg vs. (88.6 ± 12.2) mg, (57.2 ± 21.9) mg] and volume [(41.15 ± 12.17) mm³ vs. (118.45 ± 24.68) mm³, (101.36 ± 21.90) mm³] in the experimental group were significantly lower than that in the two control groups (P < 0.05). The STAT3 mRNA and protein levels of the samples from nude mice in the experimental group were significantly lower than that in the two control groups. CONCLUSION: siRNA targeting STAT3 can inhibit the proliferation of MCF7 cells in vitro and in vivo. STAT3 may become a novel therapeutic target for breast cancer.
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Neoplasias de la Mama , Proliferación Celular , Silenciador del Gen , ARN Interferente Pequeño/genética , Factor de Transcripción STAT3/genética , Animales , Apoptosis , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , ARN Mensajero/metabolismo , Factor de Transcripción STAT3/biosíntesis , Factor de Transcripción STAT3/fisiología , Transfección , Carga TumoralRESUMEN
Preeclampsia (PE) is a major obstetrical complication that results in maternal and fetal morbidity and mortality. Aberrant epigenetic modifications are widely involved in the pathogenesis of PE. Previously, the activated leukocyte cell adhesion molecule (ALCAM) was reported to be required for blastocyst implantation but has not been described in the context of pathological pregnancy. This study explored the expression of ALCAM and its methylation levels in the placentas and peripheral venous blood of patients with PE from a Chinese Han population. The mRNA and protein expression levels of ALCAM were downregulated in the PE placentas compared with the control placentas (P < 0.05). The methylation rate of the ALCAM gene promoter was considerably elevated in the placentas (P = 0.003, odds ratio (OR) = 0.264, 95% confidence interval (95% CI) [0.108-0.647], cases n = 47, controls n = 53) and peripheral blood (P = 0.007, OR = 0.455, 95% CI [0.256-0.806], cases n = 100, controls n = 100) of the PE patients compared with those of the normotensive women, suggesting a negative relationship between ALCAM methylation and gene transcription. Moreover, the transcriptional expression of ALCAM was dramatically increased by demethylating treatment in trophoblastic cells. ALCAM is expected to be involved in the pathogenesis of PE through methylation regulation.
Asunto(s)
Antígenos CD/metabolismo , Moléculas de Adhesión Celular Neuronal/metabolismo , Metilación de ADN , Proteínas Fetales/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Regiones Promotoras Genéticas , Adulto , Antígenos CD/genética , Estudios de Casos y Controles , Moléculas de Adhesión Celular Neuronal/genética , Femenino , Proteínas Fetales/genética , Humanos , Preeclampsia/genética , Embarazo , Trofoblastos/metabolismoRESUMEN
OBJECTIVE: To investigate two single nucleotide polymorphism sites of IRF5 and TLR-9 and to detect their relationship with SLE (systemic lupus erythematosus) in a Han population from Shandong province. METHODS: The polymorphisms of rs2004640 G/T, rs10954213 G/A in IRF-5 and rs187084C/T, rs352139A/G in TLR-9 were detected with PCR-RFLP in 92 cases of SLE and 88 healthy controls. The genotype and allele frequencies were calculated and analyzed. RESULTS: (1) The genotype frequencies of GG, GT and TT in rs2004640 site in SLE were 0.198, 0.521 and 0.281 respectively. The difference was significant between SLE and controls (chi(2) = 8.73, P < 0.05); the genotype frequencies of GG, GA and AA at rs109542130 site in SLE were 0.318, 0.409 and 0.273 respectively. The difference was significant between SLE and controls (chi(2) = 6.36, P < 0.05). (2) The genotype frequencies of CC, CT and TT at rs187084 site in SLE were 0.185, 0.413 and 0.402 respectively. There was no difference between SLE and controls (chi(2) = 2.99, P > 0.05); the genotype frequencies of AA, AG and GG at rs352139 site in SLE were 0.228, 0.533 and 0.239 respectively. There was no difference between SLE and controls (chi(2) = 4.54, P > 0.05). CONCLUSION: The polymorphism of rs2004640 and rs10954213 in IRF5 may be associated with SLE in the population of Han nationality from Shandong province. However, the polymorphism of rs187084 and rs352139 in TLR-9 is not associated with SLE.