Neutrófilos en el diagnóstico de infarto del miocardio tipo 4a / Neutrophils in the Diagnosis of Type 4 Myocardial Infarction

Introducción: El infarto del miocardio tipo 4a es una complicación del intervencionismo coronario percutáneo que incrementa el estado inflamatorio de los pacientes. Objetivo: Evaluar el valor diagnóstico del conteo absoluto de neutrófilos en la aparición de infarto del miocardio tipo 4a. Métodos: Se realizó una cohorte prospectiva en el Hospital Hermanos Ameijeiras. El universo estuvo constituido por 412 pacientes a los que se les realizó intervencionismo coronario percutáneo en el período comprendido de noviembre de 2018 a enero de 2021, la muestra fue de 232 pacientes. Se definieron variables clínicas, anatómicas, e inflamatorias. Resultados: Existieron diferencias significativas entre los pacientes con infarto tipo 4a y los que no tuvieron esta complicación según las variables clínicas: edad, índice de masa corporal, diabetes mellitus, enfermedad renal crónica y disfunción sistólica ventricular. La elevación del conteo absoluto de neutrófilos posterior al proceder con un área bajo la curva de 0,947 tuvo buena capacidad de discriminación de esta complicación (p = 0,000). En el diagnóstico de infarto periproceder el conteo absoluto de neutrófilos fue 7,35 posterior al proceder, tuvo una sensibilidad de 91,3 por ciento una especificidad de 96,2 por ciento. Conclusiones: Los neutrófilos fueron sensibles y específicos para el diagnóstico de infarto del miocardio tipo 4a(AU)

Introduction: Type 4 myocardial infarction is a complication of percutaneous coronary intervention that increases the inflammatory state of patients. Objective: To evaluate the diagnostic value of the absolute neutrophil count in the occurrence of type 4 myocardial infarction. Methods: A prospective cohort was carried out at Hermanos Ameijeiras Clinical Surgical Hospital. The universe consisted of 412 patients who underwent percutaneous coronary intervention from November 2018 to January 2021, two hundred thirty-two (232) patients form the sample. Clinical, anatomical and inflammatory variables were defined. Results: There were significant differences between patients with type 4 infarction and those who did not have this complication according to the clinical variables such as age, body mass index, diabetes mellitus, chronic kidney disease and ventricular systolic dysfunction. The subsequent elevation of the absolute neutrophil count when proceeding with an area under the 0.947 curve had good ability to discriminate this complication (p = 0.000). In the diagnosis of periprocedural infarction, the absolute neutrophil count was ≥ 7.35 after the procedure, it had 91.3percent sensitivity and 96.2percent specificity. Conclusions: Neutrophils were sensitive and specific for the diagnosis of type 4 myocardial infarction(AU)

Periodontite associada à artrite reumatoide: o papel das redes extracelulares de neutrófilos (NETs) / Periodontitis associated with rheumatoid arthritis: the role of neutrophil extracellular traps (NETs)

Introdução: As redes extracelulares de neutrófilos (NETs) são apontadas como um dos mecanismos relevantes na patogênese da periodontite e artrite reumatoide (AR). No entanto, permanece pouco compreendido a participação das NETs como mecanismo de ligação entre as duas doenças. Objetivos: 1) Investigar a concentração das NETs na saliva, no plasma e in vitro em indivíduos com AR e controles saudáveis e a associação com a periodontite e atividade da AR; 2) Avaliar o impacto do tratamento periodontal não cirúrgico na concentração das NETs na saliva e no plasma; 3) Investigar a associação entre a presença de polimorfismos de nucleotídeo único no gene codificador da enzima peptidil arginina deaminase 4 (PAD4) com a AR, a produção de NETs in vitro e a periodontite; 4) Sistematizar as evidências disponíveis na literatura sobre o efeito do tratamento periodontal não cirúrgico sobre os principais parâmetros clínicos e laboratoriais da AR e score de atividade da doença 28 (DAS28). Material e Métodos: Para atender aos objetivos 1 e 2, a concentração de NETs na saliva, no plasma e na cultura de neutrófilos isolados do sangue periférico foi determinada por meio da identificação do complexo mieloperoxidase (MPO)-DNA com o uso do kit PicoGreen®. Para atender ao objetivo 3 foi realizada a extração do DNA genômico das células mononucleares do sangue periférico de indivíduos com AR e controles e foi realizada a genotipagem para os polimorfismos de nucleotídeo único PADI4_89, PADI4_90, PADI4_92 e PADI_104. Para atender ao objetivo 4 foi realizada uma overview incluindo revisões sistemáticas que avaliaram o efeito do tratamento periodontal não cirúrgico sobre os parâmetros da AR. A busca foi realizada nas principais bases de dados, sem restrição de idioma ou data de publicação. Foi realizada ainda uma meta-análise incluindo dados dos estudos primários identificados nas revisões sistemáticas analisadas. Resultados: 1) e 2) Indivíduos com AR e com periodontite apresentaram maior concentração de NETs na saliva, no plasma e in vitro. O tratamento periodontal não cirúrgico reduziu a concentração de NETs na saliva e plasma de indivíduos com AR. 3) Não foi observada associação entre a presença de genótipos polimórficos e a AR. A presença de um haplótipo homozigoto para o polimorfismo foi associada a uma maior produção de NETs in vitro e piores parâmetros periodontais. 4) Foram incluídas na overview nove revisões sistemáticas. Os principais desfechos avaliados foram DAS28; proteína C-Reativa e/ou velocidade de hemossedimentação. A meta-análise mostrou que o tratamento periodontal não cirúrgico resultou em diminuição significativa do DAS28. Conclusão: A concentração das NETs na saliva, no plasma e na cultura de neutrófilos de sangue periférico está associada a AR e a periodontite, podendo representar o elo entre as duas doenças. O tratamento periodontal não cirúrgico leva à redução da atividade da AR. Polimorfismos no gene PADI4 estão associados a produção de NETs in vitro e à presença de periodontite.

Introduction: Neutrophil extracellular traps (NETs) are recognized as one of the relevant mechanisms in the pathogenesis of periodontitis and rheumatoid arthritis (RA). However, the participation of NETs as a linking mechanism between the two diseases remains poorly understood. Objectives: 1) To investigate the concentration of NETs in saliva, plasma, and in vitro in individuals with RA and healthy controls and the association of NETs with periodontitis and RA activity; 2) To evaluate the impact of non-surgical periodontal treatment on the concentration of NETs in saliva and plasma; 3) To investigate the association between the presence of single nucleotide polymorphisms in the gene coding for the enzyme peptidyl arginine deaminase 4 (PAD4) with RA, in vitro production of NETs, and periodontitis; 4) To systematize the evidence available in the literature on the effect of non-surgical periodontal treatment on the main clinical and laboratory parameters of RA and disease activity score 28 (DAS28). Material and Methods: To accomplish Objectives 1 and 2, the concentration of NETs in saliva, plasma, and culture of neutrophils isolated from peripheral blood was determined by identifying the myeloperoxidase (MPO)-DNA complex using the PicoGreen kit®. To accomplish Objective 3, genomic DNA was extracted from peripheral blood mononuclear cells of individuals with RA and healthy controls, and genotyping was performed for single nucleotide polymorphisms PADI4_89, PADI4_90, PADI4_92, and PADI_104. To accomplish the Objective 4, an overview, including systematic reviews that evaluated the effect of non-surgical periodontal treatment on RA parameters, was performed. The search was carried out in the main databases, with no restriction on language or date of publication. A meta- analysis, including data from the primary studies identified in the analyzed systematic reviews, was also performed. Results: For Objectives 1 and 2, individuals with RA and periodontitis showed a higher concentration of NETs in saliva, plasma, and in vitro. Non-surgical periodontal treatment reduced the concentration of NETs in saliva and plasma of individuals with RA. For Objective 3, no association between the presence of polymorphic genotypes and RA was observed. The presence of a homozygous haplotype for the polymorphism was associated with a higher production of NETs in vitro and worse periodontal parameters. For Objective 4, nine systematic reviews were included in the overview. The main outcomes evaluated were DAS28, C-Reactive protein, and/or erythrocyte sedimentation rate. The meta- analysis showed that non-surgical periodontal treatment resulted in a significant decrease in DAS28. Conclusion: The concentration of NETs in saliva, plasma and culture of peripheral blood neutrophils is associated with RA and periodontitis and may represent the link between the two diseases. Non-surgical periodontal treatment leads to reduced RA activity. Polymorphisms in the PADI4 gene are associated with the in vitro production of NETs and with presence of periodontitis.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5–16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5–16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios [...].

PAI-1 genetic polymorphisms influence septic patients' outcomes by regulating neutrophil activity / 中华医学杂志(英文版)

BACKGROUND@#Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the pathophysiology of sepsis, but the exact mechanism remains debatable. In this study, we investigated the associations among the serum levels of PAI-1, the incidence of 4G/5G promoter PAI-1 gene polymorphisms, immunological indicators, and clinical outcomes in septic patients.@*METHODS@#A total of 181 patients aged 18-80 years with sepsis between November 2016 and August 2018 in the intensive care unit in the Xinhua Hospital were recruited in this retrospective study, with 28-day mortality as the primary outcome. The initial serum level of PAI-1 and the presence of rs1799768 single nucleotide polymorphisms (SNPs) were examined. Univariate logistic regression and multivariate analyses were performed to determine the factors associated with different genotypes of PAI-1, serum level of PAI-1, and 28-day mortality.@*RESULTS@#The logistic analysis suggested that a high serum level of PAI-1 was associated with the rs1799768 SNP of PAI-1 (4G/4G and 4G/5G) (Odds ratio [OR]: 2.49; 95% confidence interval [CI]: 1.09, 5.68). Furthermore, a high serum level of PAI-1 strongly influenced 28-day mortality (OR 3.36; 95% CI 1.51, 7.49). The expression and activation of neutrophils (OR 0.96; 95% CI 0.93, 0.99), as well as the changes in the expression patterns of cytokines and chemokine-associated neutrophils (OR: 1.00; 95% CI: 1.00, 1.00), were both regulated by the genotype of PAI-1.@*CONCLUSIONS@#Genetic polymorphisms of PAI-1 can influence the serum levels of PAI-1, which might contribute to mortality by affecting neutrophil activity. Thus, patients with severe sepsis might clinically benefit from enhanced neutrophil clearance and the resolution of inflammation via the regulation of PAI-1 expression and activity.

Delineating asthma according to inflammation phenotypes with a focus on paucigranulocytic asthma / 中华医学杂志(英文版)

Asthma is characterized by chronic airway inflammation and airway hyper-responsiveness. However, the differences in pathophysiology and phenotypic symptomology make a diagnosis of "asthma" too broad hindering individualized treatment. Four asthmatic inflammatory phenotypes have been identified based on inflammatory cell profiles in sputum: eosinophilic, neutrophilic, paucigranulocytic, and mixed-granulocytic. Paucigranulocytic asthma may be one of the most common phenotypes in stable asthmatic patients, yet it remains much less studied than the other inflammatory phenotypes. Understanding of paucigranulocytic asthma in terms of phenotypic discrimination, distribution, stability, surrogate biomarkers, underlying pathophysiology, clinical characteristics, and current therapies is fragmented, which impedes clinical management of patients. This review brings together existing knowledge and ongoing research about asthma phenotypes, with a focus on paucigranulocytic asthma, in order to present a comprehensive picture that may clarify specific inflammatory phenotypes and thus improve clinical diagnoses and disease management.

Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation / 中国血吸虫病防治杂志

OBJECTIVE@#To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM).@*METHODS@#Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA.@*RESULTS@#Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells.@*CONCLUSIONS@#CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.

Study on the prognostic influencing factors of esophageal squamous cell carcinoma and the predictive value of inflammatory reaction indexes on its postoperative recurrence / 中华肿瘤杂志

Objective: To explore the influence factors of poor prognosis of esophageal squamous cell carcinoma (ESCC) and the predictive value of inflammatory reaction indexes including neutrophils and lymphocytes ratio (NLR), platelet and lymphocyte ratio (PLR), monocyte and lymphocyte ratio (MLR) provision and differentiation degree, infiltration depth, lymph node metastasis number on the postoperative recurrence of ESCC. Methods: A total of 130 patients with ESCC who underwent radical resection from February 2017 to February 2019 in Nanyang Central Hospital were selected and divided into good prognosis group (66 cases) and poor prognosis group (64 cases) according to the prognostic effect. The clinical data and follow-up data were collected. Multivariate logistic regression analysis was used to determine the independent influencing factors of poor prognosis. Spearman correlation analysis was used to determine the correlation between preoperative NLR, PLR and MLR with the degree of differentiation, depth of invasion and number of lymph node metastases. Receiver operating characteristic (ROC) curve analysis was used to evaluate the efficacy of NLR, PLR and MLR in predicting poor prognosis of ESCC. Results: Univariate analysis showed that the degree of differentiation, the degree of invasion and the number of lymph node metastasis were related to the prognoses of patients with ESCC (P<0.05). Multivariate logistic regression analysis showed that the degree of differentiation, depth of invasion and number of lymph node metastases were independent influencing factors for poor prognosis of patients with ESCC, moderate differentiation (OR=2.603, 95% CI: 1.009-6.715) or low differentiation (OR=9.909, 95% CI: 3.097-31.706), infiltrating into fibrous membrane (OR=14.331, 95% CI: 1.333-154.104) or surrounding tissue (OR=23.368, 95% CI: 1.466-372.578), the number of lymph node metastases ≥ 3 (OR=9.225, 95% CI: 1.693-50.263) indicated poor prognosis. Spearman correlation analysis showed that NLR was negatively correlated with the degree of differentiation and the number of lymph node metastases (r=-0.281, P=0.001; r=-0.257, P=0.003), PLR was negatively correlated with the degree of differentiation, depth of invasion and number of lymph node metastasis (r=-0.250, P=0.004; r=0.197, P=0.025; r=-0.194, P=0.027), MLR was positively correlated with the degree of differentiation and the number of lymph node metastasis (r=0.248, P=0.004; r=0.196, P=0.025). ROC curve analysis showed that the areas under the curve of NLR, PLR and MLR in predicting poor prognosis of ESCC were 0.971, 0.925 and 0.834, respectively. The best cut-off value of NLR was 2.87. The sensitivity and specificity of NLR in predicting poor prognosis of ESCC were 90.6% and 87.9%, respectively. The optimal cut-off value of PLR was 141.75. The sensitivity and specificity for predicting poor prognosis of ESCC were 92.2% and 87.9%, respectively. The best cut-off value of MLR was 0.40. The sensitivity and specificity of MLR in predicting poor prognosis of esophageal squamous cell carcinoma were 54.7% and 100.0%, respectively. Conclusions: The degree of differentiation, the degree of invasion and the number of lymph node metastases are closely related to the poor prognosis of patients with esophageal squamous cell carcinoma. NLR, PLR and MLR can provide important information for predicting the poor prognosis of esophageal squamous cell carcinoma.

Diagnostic and prognostic value of neutrophil CD64, IL-6, and PCT in ICU patients with sepsis / 中华危重病急救医学

OBJECTIVE@#To explore the significance of neutrophil CD64 (nCD64), interleukin-6 (IL-6), and procalcitonin (PCT) alone and in combination in the diagnosis and prognosis evaluation of patients with sepsis.@*METHODS@#A prospective study was conducted. Adult patients admitted to the Western Intensive Care Unit (ICU) of Yantai Yuhuangding Hospital Affiliated to Medical College of Qingdao University from September 2020 to October 2021 were selected as subjects. The venous blood of the selected patients was collected within 6 hours of entering the ICU to measure the levels of nCD64, IL-6 and PCT. The levels of nCD64, IL-6 and PCT of septic patients were measured again on the 3rd and 7th days after admission to ICU. According to the diagnostic criteria of Sepsis-3, the patients were divided into sepsis group and non-sepsis group to evaluate the diagnostic value of nCD64, IL-6 and PCT in sepsis. The patients with sepsis were divided into sepsis group and septic shock group according to their condition on admission to ICU, and the evaluation value of three biomarkers for sepsis was evaluated. Sepsis patients were divided into the survival group and the death group according to survival after 28 days, and the relationship between the three biomarkers and the prognosis of sepsis was evaluated.@*RESULTS@#Finally, 47 patients with sepsis, 43 patients with septic shock, and 41 patients without sepsis were enrolled. Seventy-six patients with sepsis survived and 14 died after 28 days. (1) The levels of nCD64, IL-6 and PCT on the first day of ICU admission in the sepsis group were significantly higher than those in the non-sepsis group [nCD64: 26.95 (14.05, 86.18) vs. 3.10 (2.55, 5.10), IL-6 (ng/L): 93.45 (52.73, 246.30) vs. 34.00 (9.76, 62.75), PCT (μg/L): 6.63 (0.57, 68.50) vs. 0.16 (0.08, 0.35), all P < 0.01]. The receiver operator characteristic curve (ROC curve) showed that the area under curve (AUC) of nCD64, IL-6 and PCT in the diagnosis of sepsis were 0.945, 0.792 and 0.888, respectively. The diagnostic value of nCD64 was the highest. When the cut-off value of nCD64 was 7.45, the sensitivity and specificity were 92.2% and 95.1% respectively. When nCD64, IL-6 and PCT were diagnosed in pairs or in combination, the combination of the three had the highest diagnostic performance, with AUC of 0.973, sensitivity of 92.2% and specificity of 97.6%. (2) The levels of nCD64, IL-6 and PCT in septic shock group were higher than those in sepsis group on the 1st, 3rd and 7th day after admission to ICU. ROC curve analysis showed that nCD64, IL-6 and PCT had some accuracy in evaluating the severity of sepsis on the 1st, 3rd and 7th day after entering ICU, with AUC ranging from 0.682 to 0.777. (3) The levels of nCD64, IL-6 and PCT in the death group were significantly higher than those in the survival group. Except for the nCD64 and PCT values on the first day after admission to the ICU, there were significant differences between the two groups in all indicators at other time points. ROC curve analysis showed that the AUC of nCD64, IL-6 and PCT predicting the prognosis of sepsis at each time point ranged from 0.600 to 0.981. The clearance rates of nCD64, IL-6 and PCT at 3 and 7 days after ICU admission were calculated by dividing the difference between the 1st and 3rd or 7th days after entering the ICU by the value at the 1st day. Logistic regression was used to analyze their predictive value for the prognosis of sepsis. The results showed that the clearance rates of nCD64, IL-6 and PCT on the 3rd and 7th day of ICU were protective factors for 28-day mortality in patients with sepsis, except the clearance rate of IL-6 on the 7th day.@*CONCLUSIONS@#nCD64, IL-6 and PCT have good value as biomarkers for the diagnosis of sepsis. The diagnostic value of nCD64 is higher than that of PCT and IL-6. When they are used together, the diagnostic value is the highest. nCD64, IL-6 and PCT have certain value in evaluating the severity and predicting the prognosis of patients with sepsis. The higher the clearance rate of nCD64, IL-6 and PCT, the lower the 28-day mortality risk of patients with sepsis.

A YAP/TAZ-CD54 axis is required for CXCR2-CD44- tumor-specific neutrophils to suppress gastric cancer

As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.

Predictive Value of Complete Blood Count and Inflammation Marker on Risk of Recurrence in Children with Henoch-Schönlein Purpura / 中国实验血液学杂志

OBJECTIVE@#To investigate the predictive value of complete blood count (CBC) and inflammation marker on the recurrence risk in children with Henoch-Schönlein purpura (HSP).@*METHODS@#One hundred and thirty-three children with HSP admitted to Cangzhou Central Hospital from February 2017 to March 2019 were enrolled. The clinical data of the children were collected, at the time of admission CBC and C-reactive protein (CRP) were detected. After discharge, the children were followed up for 1 year, the clinical data of children with and without recurrence were compared, and multivariate logistic regression was used to analyze the risk factors affecting HSP recurrence. Receiver operating characteristic (ROC) curve should be drawn and the predictive value of CBC and CRP on HSP recurrence should be analyzed.@*RESULTS@#In the follow-up of 133 children, 8 cases were lost and 39 cases recurred, with a recurrence rate of 31.20% (39/125). The age, skin rash duration, proportion of renal damage at the initial onset, percentage of neutrophils, percentage of lymphocytes, platelet count (PLT), mean platelet volume (MPV) and neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), MPV/PLT ratio (MPR), and CRP level of patients with recurrence were statistically different from those without recurrence (P <0.05). Multivariate logistic regression analysis showed that long skin rash duration, renal damage at the initial onset, increased PLR, high PLT, increased MPV and elevated CRP level were independent risk factors for recurrence in children with HSP (P <0.05). The ROC curve analysis showed that the area under the curve (AUC) of the combination of the four blood and inflammation marker (PLT, MPV, PLR and CPR) in the early prediction of HSP recurrence was 0.898, which was higher than the initial renal damage (AUC=0.687) and persistent skin rash time (AUC=0.708), with a sensitivity of 84.62% and a specificity of 83.72%.@*CONCLUSION@#Observation of CBC and CPR can predict the risk of HSP recurrence early and guide early clinical intervention.

Influence of Pre-treatment Lymphocyte/Monocyte Ratio and Neutrophil/Lymphocyte Ratio on the Prognosis of Patients with Extranodal NK/T-Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To explore the influence of lymphocyte-to-monocyte ratio (LMR) and neutrophil-to-lymphocyte ratio (NLR) on the prognosis of patients with extranodal NK/T cell lymphoma (ENKTL).@*METHODS@#The clinical data of 203 patients with ENKTL admitted to the First Affiliated Hospital of Zhengzhou University from January 2011 to January 2020 were retrospectively analyzed. The ROC curve determined the limit values of LMR and NLR; Categorical variables were compared using a chi-square test, expressed as frequency and percentage (n,%). Continuous variables were expressed as medians and extremes and compared with the Mann-Whitney U test; Progression-free survival (PFS) and overall survival (OS) of different grouped LMR and NLR patients were analyzed using Kaplan-Meier curves and compared with log-rank tests. The COX proportional risk regression model was used to perform one-factor and multi-factor analysis of PFS and OS.@*RESULTS@#The optimal critical values of LMR and NLR were determined by the ROC curve, which were 2.60 and 3.40, respectively. LMR≤2.60 was more likely to occur in patients with bone marrow invasion (P=0.029) and higher LDH (P=0.036), while NLR≥3.40 was more likely to occur in patients with higher ECOG scores (P=0.002), higher LDH (P=0.008), higher blood glucose (P=0.024), and lower PLT (P=0.010). Kaplan-Meier survival analysis showed that PFS and OS of patients in the high LMR group were significantly better than the low LMR group, while PFS and OS in the low NLR group were significantly better than the high NLR group. The results of multivariate COX analysis showed that EBV-DNA positive (P=0.047), LMR≤2.60 (P=0.014), NLR≥3.40 (P=0.023) were independent risk factors affecting PFS in patients with ENKTL. LMR≤2.60 (P<0.001), NLR≥3.40 (P=0.048), and high β2-MG (P=0.013) were independent risk factors affecting OS in patients with ENKTL.@*CONCLUSION@#Low LMR and high NLR before treatment are associated with poor prognosis in patients with ENKTL, which also can be used as an easily testable, inexpensive, and practical prognostic indicator in the clinic.

Recent research on platelet-leukocyte aggregates and their role in the pathogenesis of Kawasaki disease / 中国当代儿科杂志

Activated platelets may interact with various types of leukocytes such as monocytes, neutrophils, dendritic cells, and lymphocytes, trigger intercellular signal transduction, and thus lead to thrombosis and synthesis of massive inflammatory mediators. Elevated levels of circulating platelet-leukocyte aggregates have been found in patients with thrombotic or inflammatory diseases. This article reviews the latest research on the formation, function, and detection methods of platelet-leukocyte aggregates and their role in the onset of Kawasaki disease, so as to provide new ideas for studying the pathogenesis of Kawasaki disease.

Research progress on the relationship between low-density neutrophils and infectious diseases / 细胞与分子免疫学杂志

Neutrophils play an important role in infectious diseases by clearing pathogens in the early stages of the disease and damaging the surrounding tissues along with the disease progress. Low-density neutrophils (LDNs) are a crucial and distinct subpopulation of neutrophils. They are a mixture of activated and degranulated normal mature neutrophils and a considerable number of immature neutrophils prematurely released from the bone marrow. Additionally, they may be involved in the occurrence and development of diseases through the changes in phagocytosis, the generation of reactive oxygen species (ROS), the enhancement of the ability to produce neutrophils extracellular traps and immunosuppression. We summarizes the role of LDNs in the pathogenesis and their correlation with the severity of infectious diseases such as COVID-19, severe fever with thrombocytopenia syndrome (SFTS), AIDS, and tuberculosis.

Increased autophagy of peripheral blood neutrophils and neutrophils extracellular traps formation in systemic lupus erythematosus / 细胞与分子免疫学杂志

Objective To explore the role of autophagy, apoptosis of neutrophils and neutrophils extracellular traps (NET) formation in systemic lupus erythematosus (SLE). Methods Thirty-six patients with SLE were recruited as research subjects, and 32 healthy controls matched accordingly were enrolled as control subjects. The expression levels of microtubule associated protein 1 light chain 3B (LC3B), autophagy-related gene5(ATG5), P62, B-cell lymphoma 2(Bcl2), Bcl2-related X protein (BAX) in neutrophils were detected by Western blot analysis. Flow cytometry was employed to analyze the expression of LC3B on neutrophils. The expression level of myeloperoxidase(MPO) in plasma was estimated by ELISA. Furthermore, neutrophils were cultured in vitro and stimulated by 100 nmol/L rapamycin and 10 μg/mL lipopolysaccharide (LPS) for 6 hours, respectively. And then, the expression levels of LC3B, ATG5, P62, Bcl2 and BAX in neutrophils were detected by Western blot analysis. The level of MPO in culture supernatant was detected by ELISA. The change of fluorescence intensity of NET in culture supernatant was assayed by SytoxTM Green staining combined with fluorescence spectrophotometry. Results Compared with healthy controls, the levels of autophagy and apoptosis of neutrophils and NET formation in SLE patients were increased. The level of apoptosis and NET formation was positively associated with neutrophil autophagy. The level of autophagy showed an increase but had no effect on apoptosis and NET formation for neutrophil stimulated by rapamycin. The levels of autophagy and NET formation also increased with no significant effect on apoptosis for neutrophil induced by LPS. Conclusion The autophagy, apoptosis and NET formation of neutrophils increase in SLE patients. The activation of autophagy and NET in neutrophils possibly result from the inflammatory internal environment in SLE patients.

Accelerating Wound Healing of Traumatic Ulcer with Topical Application of Dental Pulp Mesenchymal Stem Cell Secretome and Robusta Green Coffee Bean Extract Combination in vivo

ABSTRACT Objective: To investigate the ability of a combination dental pulp mesenchymal stem cell secretome (DPMSCS), robusta green coffee bean extract (RGCBE), and Carboxymethylcellulose-Natrium (CMC-Na) in a Wistar rats model of traumatic ulcers. Material and Methods: Twenty-eight young, male, healthy Wistar rats (Rattus norvegicus) were divided into seven groups randomly: Group K0, group K1-3 (traumatic ulcer rats that received CMC-Na gel for three days), group K1-7 (traumatic ulcer rats that received CMC-Na gel for seven days), group K2-3 (traumatic ulcer rats that received RGCBE for three days), group K2-7 (traumatic ulcer rats that received RGCBE for seven days), group K3-3 (traumatic ulcer rats that received DPMSCS for three days), and group K3-7 (traumatic ulcer rats that received DPMSCS for seven days). An ulcer was made with an amalgam stopper on the right buccal mucosa of the rats. DPMSCS 50% gel was applied to the ulcer on the left buccal mucosa. The ulcer diameter was measured on day 3 and day 7. Results: There was a significant difference in the diameter of the ulcer, the number of neutrophils, and fibroblasts in the treatment group compared to the control group on day 7. Conclusion: A combination of DPMSCS and RGCBE 50% accelerates traumatic ulcer wound healing by lowering ulcer diameter, decreasing neutrophil counts, and increasing fibroblast proliferation in vivo.

Can NLR be a biomarker for mucositis and gvhd in patients undergoing allogeneic HSCT?

Allogeneic hematopoietic stem cell transplantation (HSCT) is a treatment for many diseases; however, it can induce complications such as Oral Mucositis (OM) and Graft-versus- Host Disease (GVHD). The neutrophil-lymphocyte ratio (NLR) is a peripheral biomarker of systemic inflammation and an independent prognostic factor for several inflammatory diseases. Aim: This study aimed to evaluate the association of NLR with OM and GVHD in patients undergoing allogeneic HSCT. Methods: Patients who underwent allogeneic HSCT at the Bone Marrow Transplant Service of the Hospital de Clínicas Complex of the Federal University of Paraná were included in the study. Socio-demographic data and blood counts were collected from patients' medical records. The NLR was calculated and associated with OM and GVHD. Results: 45 patients were included in the study. Although NLR was higher in patients with OM and oral GVHD, no statistical difference was observed, and no relationship between OM and GVHD with NLR could be stated. Conclusion: Although both OM and GVHD are associated with an inflammatory response as well as the immune system, it was not associated with NLR. Further investigation considering other variables related to HSCT might find possible associations, as it could favor patient management and prevention

Índice neutrófilos-linfocitos en el diagnóstico de apendicitis aguda. Una evaluación de su precisión diagnóstica / Neutrophil-to-lymphocyte ratio in the diagnosis of acute appendicitis. Assessment of its diagnostic accuracy

Introducción. La apendicitis aguda (AA) en pacientes pediátricos requiere de un diagnóstico certero. El índice neutrófilos-linfocitos (INL) es un parámetro accesible que puede ser útil en su diagnóstico. Objetivo. Determinar la precisión del INL para diagnosticar AA en pacientes con dolor abdominal. Población y métodos. Estudio de prueba diagnóstica. Se incluyeron 520 pacientes atendidos en el servicio de urgencias pediátricas. Para cuantificar la precisión diagnóstica, se estimó la sensibilidad, la especificidad, los valores predictivos (VP) y los cocientes de probabilidad (CP). Se utilizó un modelo de regresión logística múltiple para evaluar el efecto de las potenciales variables confusoras en la relación entre el INL y la AA. Resultados. La prevalencia de AA fue del 49 %. Para un punto de corte de 5, la sensibilidad fue del 85,1 %, especificidad: 78,9 %, VP+: 79,5 % y VP-: 84,6 %. Sin embargo, basándose en los cocientes de probabilidad, el INL es una prueba poco potente para el diagnóstico de AA (CP+ = 4,03 y CP- = 0,18) y resultó una prueba sin utilidad diagnóstica en el caso de apendicitis complicada (CP+ = 1,57 y CP- = 0,55). Después del ajuste por edad, sexo, obesidad, tiempo de evolución y uso de analgésicos, el INL fue una variable explicativa de la presencia de AA (odds ratio = 23,53; IC95 % 13,14-42,15). Conclusiones. El INL no es lo suficientemente preciso aisladamente para confirmar o descartar la presencia de AA. No obstante, el INL puede emplearse junto con otras pruebas para seleccionar a los pacientes en los cuales es necesario un mayor estudio.

Introduction. Acute appendicitis (AA) in pediatric patients requires an accurate diagnosis. The neutrophil-to-lymphocyte ratio (NLR) is an accessible parameter useful for its diagnosis. Objective. To determine NLR accuracy to diagnose AA in patients with abdominal pain. Population and methods. Diagnostic test study. A total of 520 patients seen at the Pediatric Emergency Department were included. Diagnostic accuracy was estimated based on sensitivity, specificity, predictive values, and likelihood ratios. A multiple logistic regression model was used to assess the effect of potentially confounding variables in the relationship between NLR and AA. Results. The prevalence of AA was 49%. For a cutoff point of 5, sensitivity was 85.1%, specificity: 78.9%; positive predictive value: 79.5%; and negative predictive value: 84.6%. However, based on likelihood ratios, the NLR is not powerful enough to diagnose AA (positive likelihood ratio = 4.03 and negative likelihood ratio = 0.18) and did not exhibit diagnostic usefulness in complicated appendicitis (positive likelihood ratio = 1.57 and negative likelihood ratio = 0.55). Following adjustment for age, sex, obesity, time since symptom onset, and analgesic use, the NLR was an explanatory variable for the presence of AA (odds ratio = 23.53; 95% confidence interval: 13.14­42.15). Conclusions. The NLR alone is not sufficiently accurate to confirm or rule out the presence of AA. However, the NLR can be used together with other tests to select patients in whom further study is necessary.

Perfiles hematológicos en pacientes infectados con malaria en un área endémica del Perú / Hematological profiles of malaria-infected patients in an endemic area of Peru

RESUMEN Objetivos. Evaluar la variación de los perfiles hematológicos antes, durante y después del tratamiento de pacientes infectados con malaria no complicada por Plasmodium vivax (Pv) y P. falciparum (Pf) en una población de la región Loreto. Materiales y métodos. El estudio se realizó entre 2010 y 2012, en Zungarococha (Iquitos). Los 425 participantes tuvieron tres visitas (visita 1-día 0-antes del tratamiento, visita 2-día 7-durante tratamiento, visita 3-día 28-después del tratamiento), hemograma completo, diagnóstico microscópico y molecular (PCR). Resultados. En la primera visita, se encontraron 93 (21,9%) positivos a Pv y 34 (8,0%) a Pf. Todos los positivos mostraron una reducción en los indicadores hematológicos de hematocrito, recuento de glóbulos blancos (RGB), neutrófilos abastonados y segmentados, eosinófilos y plaquetas (p<0.001) en comparación con el grupo negativo. Se encontró un porcentaje mayor de neutrófilos abastonados en Pf y de neutrófilos segmentados en Pv comparado al grupo negativo. Se observó variaciones en los perfiles hematológicos después del tratamiento para ambas especies, los neutrófilos abastonados disminuyeron, las plaquetas aumentaron, los eosinófilos se incrementaron al día 7 y decaen el día 28, el hematocrito y los neutrófilos segmentados disminuyeron al día 7 y se normalizaron el día 28. Las diferencias entre especies en el tiempo mostraron una disminución diaria de neutrófilos abastonados en infectados con Pv que en Pf. Conclusiones. El perfil hematológico en pacientes positivos a malaria no complicada varía en el tiempo durante y después del tratamiento. Estos son indicadores de la progresión de la enfermedad y ayudan en la vigilancia terapéutica de pacientes infectados con Plasmodium.

ABSTRACT Objectives. To evaluate the variation of hematological profiles of patients infected with uncomplicated Plasmodium vivax (Pv) and P. falciparum (Pf) malaria before, during and after treatment in a population of the Loreto region. Materials and methods. This study was conducted between 2010 and 2012, in Zungarococha (Iquitos). The 425 participants had three visits (visit 1-day 0-before treatment, visit 2-day 7-during treatment, visit 3-day 28-after treatment), complete blood count, microscopic and molecular diagnosis (PCR). Results. At the first visit, 93 (21.9%) participants were found positive for Pv and 34 (8.0%) for Pf. All positives showed a reduction in hematocrit, white blood cell count (WBC), ablated and segmented neutrophils, eosinophils and platelets (p<0.001) compared to the negative group. A higher percentage of ablated neutrophils was found in Pf and segmented neutrophils in Pv compared to the negative group. Variations in hematological profiles were observed after treatment for both species; ablated neutrophils decreased, platelets increased, eosinophils increased at day 7 and declined at day 28, hematocrit and segmented neutrophils decreased at day 7 and normalized at day 28. Interspecies differences over time showed a bigger daily decrease in ablated neutrophils in Pv-infected when compared to Pf. Conclusions. The hematological profile in uncomplicated malaria-positive patients varies over time during and after treatment. These are indicators of disease progression and help in the therapeutic surveillance of Plasmodium-infected patients.

Morphological changes in leukocytes of acute SARS-CoV-2 infection patients, Amazon, Brazil / Alterações morfológicas em leucócitos de pacientes com infecção aguda por SARS-CoV-2, Amazônia, Brasil

Introduction: The pandemic for the new coronavirus (SARS-CoV-2) brought many uncertainties about which laboratory parameters would be most suitable during the evolution of COVID 19. Objectives: Correlate the results of the blood count (BC), the neutrophil/lymphocyte ratio (N/LR), the C-reactive protein (CRP) and morphological findings of individuals diagnosed with SARS-CoV-2 infection through Polymerase Chain Reaction in Real Time (RT-PCR) in a private laboratory in Belém, Pará, from March to September 2020. Materials and Methods: Retrospective study with 30 individuals, of both sexes, any age and clinical complaint, of home or hospital origin who underwent BC, CRP and RT-PCR for COVID 19 until the 8th day of infection. Morphological changes were analyzed after selecting the slides for these patients. Results: Sample composed of 15 men and 15 women, aged between 7 and 92 years. Of these 12/30 individuals were at home and 18/30 were hospitalized. The main complaints were fever, malaise, diarrhea and respiratory distress. The statistical study showed a direct dependency relationship between increases in N/LR, CRP and the need for hospitalization (p = 0.0005). Morphological analysis showed hyposegmented neutrophils with toxic granulations, vacuolated monocytes, and reactive lymphocytes with basophilic cytoplasm. Conclusion: Our results associate intermediate and elevated levels of N/LR with increased CRP and disease severity, however, unrelated to the morphological findings in neutrophils, lymphocytes and monocytes that were common to all patients diagnosed up to the 8th day of infection (AU)

Introdução: A pandemia pelo novo coronavírus (SARS-CoV-2) trouxe muitas incertezas sobre quais parâmetros laboratoriais seriam mais adequados durante a evolução da COVID 19. Objetivos: Correlacionar os resultados do hemograma (HGM), da relação neutrófilos/linfócitos (R N/L), da proteína C reativa (PCR) e dos achados morfológicos de indivíduos diagnosticados com infecção por SARS-CoV-2 através de Reação em Cadeia da Polimerase em Tempo Real (RT-PCR) em um laboratório particular de Belém, Pará, no período de março a setembro de 2020. Materiais e Métodos: Estudo retrospectivo com 30 indivíduos, de ambos os sexos, qualquer idade e queixa clínica, de origem domiciliar ou hospitalar que realizaram HGM, PCR e RT-PCR para COVID 19 até o 8o dia de infecção. As alterações morfológicas foram analisadas após a seleção das lâminas desses pacientes. Resultados:Amostra composta por 15 homens e 15 mulheres, com idades entre 7 e 92 anos. Desses, 12/30 indivíduos estavam em domicílio e 18/30 internados. As principais queixas foram febre, mal-estar geral, diarreia e desconforto respiratório. O estudo estatístico mostrou a existência de relação de dependência direta entre os aumentos da R N/L, PCR e necessidade de internação (p=0,0005). A análise morfológica mostrou neutrófilos hipossegmentados com granulações tóxicas, monócitos vacuolizados e linfócitos reativos com citoplasma basofílico. Conclusão: Nossos resultados associam os níveis intermediários e elevados da R N/L com o aumento de PCR e a gravidade da doença, porém, sem relação com os achados morfológicos em neutrófilos, linfócitos e monócitos que foram comuns a todos os pacientes diagnosticados até o 8o dia de infecção