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

Mean platelet volume and neutrophil to lymphocyte ratio in patients with tinnitus: a case-control study / Volume plaquetário médio e relação neutrófilos/linfócitos em pacientes com zumbido: estudo de caso-controle

Abstract Introduction: Different theories have been proposed on the etiology of tinnitus, including metabolic and audiologic causes. We suggest that mean platelet volume and neutrophil to lymphocyte ratio levels change in tinnitus, indicating microcirculatory disturbance and inflammatory process in the etiopathogenesis of tinnitus. Objectives: We aimed to evaluate the mean platelet volume and neutrophil to lymphocyte ratio in patients with tinnitus in comparison to healthy controls. Methods: Retrospective case-control study. Two-hundred and eighty-seven patients aged 18-59 years and diagnosed with tinnitus in the Ear, Nose, and Throat Clinic between December 2014 and May 2017 (patient group) and 275 healthy individuals who applied for a hearing screening within the same time period (control group). Demographics, concomitant diseases, laboratory results, and audiometric data were recorded. Mean platelet volume and neutrophil to lymphocyte ratio were the outcome measures. Patients with hearing loss due to presbycusis or another reasons, and patients with anatomical disorders in the external and middle ear were excluded from the study by using physical examinations, pure audio audiometry results and radiological imaging. The upper age limit was set at 59 to exclude presbycusis patients. Results: The ratio of female patients was higher in patient group than control group (58.5%, n = 168 vs. 49.4%, n= 127; respectively; p = 0.033). The mean age of patient group was significantly higher than those of control group (44.89 ± 10.96 years and 38.37 ± 10.65 years, respectively; p = 0.001). The percentage of subjects with high mean platelet volume level was significantly higher in patient group than control group (9.4%, n = 27, and 3.1%, n = 8 respectively; p = 0.008). The mean neutrophil to lymphocyte ratio was higher in patients with tinnitus than control group (1.95 ± 1.02 and 1.67 ±0.57, p = 0.012). A neutrophil to lymphocyte ratio level of 2.17 and above is associated with 1.991 times higher risk of tinnitus (odds ratio = 1.99, 95% confidence interval 1.31-3.02). Conclusion: High mean platelet volume and neutrophil to lymphocyte ratio values are associated with idiopathic tinnitus, suggesting the role of vascular pathologies in etiology of tinnitus. Tinnitus may be a sign of underlying systemic or local disorders. Therefore, patients with tinnitus should undergo detailed evaluation including hematological indices.

Resumo Introdução: Diferentes teorias já foram propostas sobre a etiologia do zumbido, inclusive causas metabólicas e audiológicas. Acreditamos que os níveis do volume plaquetário médio e da relação neutrófilos/linfócitos se alteram no zumbido, sugerem distúrbio microcirculatório e processo inflamatório na etiopatogenia do zumbido. Objetivo: Avaliar o volume plaquetário médio e a relação neutrófilos/linfócitos em pacientes com zumbido em comparação com controles saudáveis. Método: Estudo de caso-controle retrospectivo, com 287 pacientes entre 18 e 59 anos e diagnosticados com zumbido na Clínica de Otorrinolaringologia entre dezembro de 2014 e maio de 2017 (grupo pacientes) e 275 indivíduos saudáveis que solicitaram uma triagem auditiva no mesmo período (grupo controle). Foram registrados dados demográficos, doenças concomitantes, resultados laboratoriais e dados audiométricos. O volume plaquetário médio e a relação neutrófilos/linfócitos foram as medidas de desfecho. Pacientes com perda auditiva por presbiacusia ou por outros motivos e pacientes com distúrbios anatômicos na orelha externa e média foram excluídos do estudo por meio de exame físico, resultados de audiometria tonal pura e imagens radiológicas. O limite de idade superior foi fixado em 59 anos para excluir pacientes com presbiacusia. Resultados: A proporção de pacientes do sexo feminino foi maior no grupo de pacientes do que no grupo controle (58,5%, n = 168 vs. 49,4%, n = 127; respectivamente; p = 0,033). A média de idade do grupo de pacientes era significantemente maior do que a do grupo controle (44,89 ± 10,96 anos e 38,37 ± 10,65 anos, respectivamente; p = 0,001). A porcentagem de indivíduos com nível alto de volume plaquetário médio foi significantemente maior no grupo de pacientes do que no grupo controle (9,4%, n = 27 e 3,1%, n = 8, respectivamente; p = 0,008). A relação neutrófilos/linfócitos média foi maior nos pacientes com zumbido do que no grupo controle (1,95 ± 1,02 e 1,67 ±0,57, p = 0,012). Um nível de relação neutrófilos/linfócitos de 2,17 e acima está associado a um risco 1,991 vez maior de zumbido (odds ratio = 1,99, Intervalo de Confiança de 95% 1,31 a 3,02). Conclusão: Altos valores de volume plaquetário médio e relação neutrófilos/linfócitos estão associados ao zumbido idiopático, sugerem o papel de doenças vasculares na etiologia do zumbido. O zumbido pode ser um sinal de distúrbios sistêmicos ou locais subjacentes. Portanto, pacientes com zumbido devem ser submetidos a uma avaliação detalhada, inclusive índices hematológicos.

Prognostic value of pre-operative peripheral inflammation markers in patients with squamous cell carcinoma of the external auditory canal / Valor prognóstico dos marcadores de inflamação periférica pré-operatórios em pacientes com carcinoma espinocelular do meato acústico externo

Abstract Introduction: Squamous cell carcinoma of the external auditory canal is a rare entity. Previous studies have suggested predictors for tumor recurrence. However, most of the prognostic factors were from the clinicopathological aspect. Objective: This study aims to analyze the correlation between pre-operative peripheral inflammation markers and survival outcomes, in order to identify prognostic biomarkers for patients with squamous cell carcinoma of the external auditory canal. Methods: We retrospectively analyzed patients diagnosed with squamous cell carcinoma of the external auditory canal who underwent surgery at our institute. The pre-operative circulating inflammatory markers, such as the neutrophil, lymphocyte, platelet, and monocyte counts were measured and their ratios including neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio and lymphocyte-to-monocyte ratio were calculated. The prognostic value of the measured hematologic parameters in relation to the survival outcomes was also evaluated. Results: A total of 83 patients were included, of which 26 patients showed tumor recurrence and 57 without recurrence. Neutrophil counts and neutrophil-to-lymphocyte ratio were closely connected with tumor stage. In the patients with recurrence, neutrophil counts, neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio were elevated (p< 0.0001, p< 0.0001 and p = 0.001), while lymphocyte counts and lymphocyte-to-monocyte ratio were decreased (p = 0.013 and p = 0.016, respectively). The receiver operating curve analysis indicated that pre-operative neutrophil-to-lymphocyte ratio is a potential prognostic marker for recurrence of squamous cell carcinoma of the external auditory canal (area under curve = 0.816), and the cut-off points was 2.325. Conclusions: Pre-operative neutrophil and lymphocyte counts, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, lymphocyte-to-monocyte are significantly correlated with tumor recurrence in patients with external auditory canal squamous cell carcinoma. Furthermore, neutrophil-to-lymphocyte ratio may be unfavorable prognostic factors of this disease.

Resumo Introdução: O carcinoma espinocelular do meato acústico externo é uma doença rara. Estudos anteriores sugeriram preditores de recorrência do tumor. Entretanto, a maioria dos fatores prognósticos se originou do aspecto clínico-patológico. Objetivo: Analisar a correlação entre marcadores inflamatórios periféricos pré-operatórios e os desfechos de sobrevida e identificar biomarcadores prognósticos para pacientes com carcinoma espinocelular do meato acústico externo. Método: Analisamos retrospectivamente pacientes com diagnóstico de carcinoma espinocelular do meato acústico externo submetidos à cirurgia em nosso instituto. Os marcadores inflamatórios circulantes pré-operatórios, como as contagens de neutrófilos, linfócitos, plaquetas e monócitos, foram medidos e as suas relações calculadas, inclusive as relações neutrófilos/linfócitos, plaquetas/linfócitos e linfócitos/monócitos. O valor prognóstico dos parâmetros hematológicos medidos em relação aos desfechos de sobrevida também foi avaliado. Resultados: Foram incluídos 83 pacientes, entre os quais 26 apresentaram recorrência tumoral e 57 não apresentaram. A contagem de neutrófilos e a relação neutrófilo/linfócito estavam intimamente associadas ao estágio do tumor. Nos pacientes com recorrência, a contagem de neutrófilos, a relação neutrófilos/linfócitos e a relação plaquetas/linfócitos eram elevadas (p < 0,0001, p > 0,0001 e p = 0,001), enquanto a contagem de linfócitos e a relação linfócitos/monócitos estavam diminuídas (p = 0,012 ep = 0,016, respectivamente). A análise da curva, Receiver Operating Characteristic, indicou que a relação neutrófilos/linfócitos pré-operatória era um potencial marcador prognóstico para a recorrência de carcinoma espinocelular do meato acústico externo (Área sob a curva = 0,816) e o ponto de corte foi de 2,325. Conclusão: A contagem pré-operatória de neutrófilos e linfócitos, as relações neutrófilos/linfócitos, plaquetas/linfócitos e linfócitos/monócitos estão significativamente correlacionadas com a recorrência do tumor em pacientes com carcinoma espinocelular do meato acústico externo. Além disso, a relação neutrófilos/linfócitos pode ser um fator prognóstico desfavorável dessa doença.

Severity Markers of Acute Appendicitis: Diagnostic Test Study / Marcadores de severidad de la apendicitis aguda: estudio de prueba diagnóstica

Abstract Introduction: Acute appendicitis (AA) is one of the most frequent abdominal surgical pathologies globally, with appendectomy being the most performed emergency surgery. Aim: To determine potential markers of AA severity for diagnostic purposes and timely management, thus avoiding possible complications. Materials and methods: This research relies on a randomized sample of 239 patients diagnosed with AA at the Orinoquía Regional Hospital, Colombia. We analyzed blood count, C-reactive protein (CRP), and neutrophil/lymphocyte ratio (NLR) records and established their relationship with the AA surgical findings described by Guzmán-Valdivia. Results: In the emergency department, these reactants can provide an approximate diagnosis as markers of the AA severity, with CRP > 15 mg/dL (diagnostic accuracy of 76.15 %) and NLR > 85 % (diagnostic accuracy of 61.09 %) having the best initial operating performance. Regarding complications such as intestinal perforation, we found a statistical relationship; CRP > 15 mg/dL and NLR > 85 % were the markers with the highest predictive performance, with OR 14.46 and OR 2. 17, respectively, regarding Guzmán-Valdivia's findings. Conclusions: CRP and NLR > 85 % are the acute phase reactants with the best diagnostic characteristics to predict potential AA complications.

Resumen Introducción: la apendicitis aguda (AA) es una de las patologías quirúrgicas abdominales más frecuentes en el mundo, siendo la apendicectomía, la cirugía de emergencia más realizada a nivel mundial. Objetivo: determinar los posibles marcadores de severidad en la apendicitis aguda con fines diagnósticos y para el manejo oportuno de la apendicitis y, de esta manera, evitar posibles complicaciones. Metodología: esta investigación se basó de una muestra aleatorizada de un total de 239 pacientes con diagnóstico de apendicitis aguda en el Hospital Regional de Orinoquía, Colombia. Se analizaron registros de cuadro hemático, proteína C reactiva (PCR) e índice de neutrófilo/linfocito (INL), y se estableció la relación de los mismos con los hallazgos quirúrgicos de apendicitis aguda descritos por Guzmán-Valdivia. Resultados: en el servicio de urgencias, estos reactantes son capaces de realizar una aproximación en el diagnóstico y como marcadores de la severidad de la AA, siendo la PCR > 15 mg/dL (precisión diagnóstica 76,15 %) y el porcentaje de neutrófilos > 85 % (precisión diagnóstica 61,09 %) los de mejor rendimiento operativo inicial. En cuanto a las complicaciones, como la perforación intestinal, se encontró en relación estadística, que la PCR > 15 mg/dL y el porcentaje de neutrófilos > 85 % fueron los marcadores con mayor rendimiento predictivo, con OR 14,46 y OR 2,17, respectivamente. Lo anterior en relación con los hallazgos descritos por Guzmán-Valdivia. Conclusiones: la elevación de la PCR y del porcentaje de neutrófilos > 85 % son los reactantes de fase aguda que presentan mejores características diagnósticas y para predecir posibles complicaciones de la apendicitis aguda.

Usefulness of the neutrophil-to-lymphocyte ratio in predicting the severity of COVID-19 patients: a retrospective cohort study

ABSTRACT BACKGROUND: Quick and accurate identification of critically ill patients ensures appropriate and correct use of medical resources. In situations that threaten public health, like pandemics, rapid and effective methods are needed for early disease detection among critically ill patients. OBJECTIVE: To determine the relationship between the neutrophil-to-lymphocyte ratio (NLR) of coronavirus disease-19 (COVID-19) patients upon admission to the emergency department (ED) and these patients' prognosis. DESIGN AND SETTING: Retrospective cohort study among COVID-19 patients in the ED of a tertiary-level hospital. METHODS: Data on patients' age, gender, vital signs, chronic diseases, laboratory tests and clinical outcomes were collected from electronic medical records. Receiver operating characteristic (ROC) curve analysis was performed. The area under the curve (AUC) was used to assess the accuracy of NLR for predicting in-hospital mortality risk and intensive care unit (ICU) requirement. The Youden J index (YJI) was used to determine optimal threshold values. RESULTS: 1,175 patients were included. Their median age was 63 years (IQR, 48-75). With an NLR cutoff value of 5.14, the sensitivity, specificity, PPV, AUC and YJI for ICU requirement were calculated as 77.87%, 74.08%, 92.4%, 0.811 and 0.5194, respectively. With the same cutoff value, the sensitivity, specificity, AUC and YJI for in-hospital mortality were 77.27%, 75.82%, 0.815 and 0.5309, respectively. In addition, advanced age, leukocytosis, anemia and lymphopenia were found to be associated with poor prognosis. CONCLUSION: The NLR, which is a widely available simple parameter, can provide rapid insights regarding early recognition of critical illness and prognosis among COVID-19 patients.

Hemogram index parameters in the evaluation of male breast cancer and inflammatory response: a case-control study

SUMMARY OBJECTIVE: Our aim was to investigate the hemogram index parameters and their clinical significance in the evaluation of the inflammatory response of patients with male breast cancer, who are rarely observed in the literature. METHODS: In total, 22 (n=22) healthy male and 28 (n=28) male breast cancer patients without synchronous/metachronous tumors were included in this study. They were grouped as the healthy male control group (Group 1) and the male breast cancer patient group (Group 2). The male breast cancer was divided into two subgroups, namely, early stage [(stage: 0/I/II) (Group 2A)] and late stage [(stage: III/IV) (Group 2B)], and their hemogram index parameters were compared. RESULTS: A significant (p>0.05) increase was observed in neutrophil/lymphocyte ratio (NLR) and·platelet/lymphocyte ratio (PLR) values in the late stage (Group 2B: stage III/IV) compared to the early stage (Group 2A: stage 0/I/II) and healthy control (Group 1) groups. CONCLUSIONS: In male breast cancer patients, neutrophil/lymphocyte ratio and platelet/lymphocyte ratio values were significantly higher as the stage of cancer increased. These readily available simple tests can be used to evaluate the host's inflammatory response in male breast cancer.

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

RESUMO: 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. (AU)

ABSTRACT: Morphological changes in leukocytes of acute SARS-CoV-2 infection patients, Amazon, BrazilORIGINAL ARTICLEIntroduction: 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)

A map of neutrophil granules by proteomics / Mapa dos neutrófilos por proteômica (OU) Mapa dos grânulos do neutrófilo por proteômica

Neutrophils are polymorphonuclear leukocytes that play a key role in the organism defense. These cells enroll in a range of actions to ensure pathogen elimination and orchestrate both innate and adaptative immune responses. The main physiological structures of neutrophils are their storage organelles that are essential since the cells activation and participate in all their functions. The storage organelles are divided into 2 types: granules and secretory vesicles. The granules are subdivided into azurophilic, specific and gelatinase. The granules are distinguished by their protein content, and since they play an important role on the neutrophil function, the knowledge of the proteins stored in these organelles can help to better understand these cells. Some proteins are present in high abundance and are used as markers for each storage organelle. These proteins are myeloperoxidase (MPO) for azurophil granules, neutrophil gelatinase associated with lipocalin-2 (NGAL) and lactoferrin (LTF) for specific granules, matrix metalloproteinase-9 (MMP9) for gelatinase granules and alkaline phosphatase (AP) for secretory vesicles. The isolation of neutrophils granules, however, is challenging and the existing procedures rely on large sample volumes, about 400 mL of peripheral blood or 3 x 108 neutrophils, not allowing for multiple biological and technical replicates. Therefore, the aim of this study was to develop a miniaturized neutrophil granules isolation method and to use biochemical assays, mass spectrometry-based proteomics and a machine learning approach to investigate the protein content of the neutrophils storage organelles. With that in mind, 40 mL of the peripheral blood of three apparently healthy volunteers were collected. The neutrophils were isolated, disrupted using nitrogen cavitation and organelles were fractionated with a discontinuous 3-layer Percoll density gradient. The presence of granules markers in each fraction was assessed using western blot , gelatin zymography and enzymatic assays. The isolation was proven successful and allowed for a reasonable separation of all neutrophils storage organelles in a gradient of less than 1 mL, about 37 times smaller than the methodsdescribed in the literature. Moreover, mass spectrometry-based proteomics identified 369 proteins in at least 3 of the 5 samples, and using a machine learning strategy, the localization of 140 proteins was predicted with confidence. Furthermore, this study was the first to investigate the proteome of neutrophil granules using technical and biological replicates, creating a reliable database for further studies. In conclusion, the developed miniaturized method is reproducible, cheaper, and reliable. In addition, it provides a resource for further studies exploring neutrophil granules protein content and mobilization during activation with different stimuli

Neutrófilos são leucócitos polimorfonucleares que possuem papel fundamental na defesa do organismo. Essas células desempenham diversas ações a fim de assegurar a eliminação de um patógeno e, além disso, orquestram a resposta imune inata e adaptativa. O conjunto composto pelos grânulos de armazenamento e as vesículas secretórias compõe a principal estrutura fisiológica dos neutrófilos. Estes componentes são essenciais desde a ativação celular, participando de todas as funcionalidades desta célula. Os grânulos são subdivididos em azurófilos, específicos e gelatinase. Eles podem ser distinguidos por meio de seu conteúdo proteico e, como são importantes na funcionalidade dos neutrófilos, identificar quais proteínas são armazenadas nestas organelas é imprescindível para entender melhor essa célula como um todo. Algumas proteínas, estão presentes de forma abundante e, portanto, são utilizadas como marcadores dos grânulos. Tais proteínas são mieloperoxidase (MPO) para os grânulos azurófilos, gelatinase de neutrófilo associada a lipocalina (NGAL) e lactoferrina (LTF) para os específicos, metaloproteinase de matrix 9 (MMP9) para os grânulos de gelatinase e fosfatase alcalina (AP) para as vesículas secretórias. Isolar estas estruturas, no entanto, é desafiador visto que os protocolos existentes na literatura utilizam grandes volumes de amostra, cerca de 400 mL de sangue ou 3 x 108 neutrófilos, para apenas um isolamento, impedindo a realização de replicatas técnicas e biológicas. Desta forma, o objetivo do presente estudo foi desenvolver um protocolo miniaturizado de isolamento dos grânulos neutrofílicos e utilizar métodos bioquímicos, de proteômica e machine learning para investigar o conteúdo proteico destas estruturas celulares. Para isto, 40 mL de sangue periférico de três voluntários aparentemente saudáveis foi coletado. Os neutrófilos foram então isolados, lisados com cavitação de nitrogênio e o fracionamento subcelular foi realizado baseado em um gradiente descontínuo de 3 camadas de Percoll. O método de isolamento foi avaliado através da investigação dos marcadores utilizando western blotting (WB), zimografia de gelatina e ensaios enzimáticos em cada fração coletada. O isolamento demonstrou-se eficiente e permitiu uma ótima separação dos grânulosem um gradiente menor que 1 mL, cerca de 37 vezes menor que os métodos atualmente descritos na literatura. Além disso, a análise proteômica foi capaz de identificar 369 proteínas presentes em pelo menos 3 das 5 réplicas investigadas e, utilizando ferramentas de machine learning, 140 proteínas foram classificadas como pertencentes a um dos tipos de grânulos ou vesícula secretória com alto nível de confiabilidade. Por fim, o presente estudo foi o primeiro a investigar o proteoma dos grânulos utilizando replicatas técnicas e biológicas, criando e fornecendo uma base de dados robusta que poderá ser utilizada em estudos futuros. Conclui-se, portanto, que a metodologia miniaturizada desenvolvida é eficaz, reprodutível e mais barata, além de permitir estudos mais complexos e profundos sobre o proteoma dos grânulos dos neutrófilos em diferentes momentos celulares, tais como quando ativados via estímulos distintos

PFKL, a novel regulatory node for NOX2-dependent oxidative burst and NETosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Neutrophils are predominant leukocytes in the circulation, which are essential for killing invading pathogens via the activation of effector responses and the production of reactive oxygen species (ROS), also named as "oxidative burst." When infected, activated neutrophils fight bacteria, fungi, and viruses through oxidative burst, phagocytosis, degranulation, and the production of neutrophil extracellular traps (NETs) in a neutrophil death process named as "NETosis" (Mutua and Gershwin, 2021). NETs, consisting of DNA fibers decorated with modified histones and numerous antimicrobial proteins from cytoplasmic granules and the nucleus, can either be beneficial or detrimental (Mutua and Gershwin, 2021). Several pathways can lead to this death process. In response to various stimuli, NETosis traps and clears pathogens, facilitating phagocytosis by other neutrophils and phagocytes. However, excessive NETosis often results in disease due to increasing the pro-inflammatory response and perpetuating the inflammatory condition (Hellebrekers et al., 2018; Hidalgo et al., 2019; Klopf et al., 2021). Accordingly, inhibiting aberrant NETosis may alleviate the severity of various autoimmune and inflammatory diseases.

Roles of neutrophil reactive oxygen species (ROS) generation in organ function impairment in sepsis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Sepsis is a condition of severe organ failure caused by the maladaptive response of the host to an infection. It is a severe complication affecting critically ill patients, which can progress to severe sepsis, septic shock, and ultimately death. As a vital part of the human innate immune system, neutrophils are essential in resisting pathogen invasion, infection, and immune surveillance. Neutrophil-produced reactive oxygen species (ROS) play a pivotal role in organ dysfunction related to sepsis. In recent years, ROS have received a lot of attention as a major cause of sepsis, which can progress to severe sepsis and septic shock. This paper reviews the existing knowledge on the production mechanism of neutrophil ROS in human organ function impairment because of sepsis.

Design, simulation and application of multichannel microfluidic chip for cell migration / 生物医学工程学杂志

Cell migration is defined as the directional movement of cells toward a specific chemical concentration gradient, which plays a crucial role in embryo development, wound healing and tumor metastasis. However, current research methods showed low flux and are only suitable for single-factor assessment, and it was difficult to comprehensively consider the effects of other parameters such as different concentration gradients on cell migration behavior. In this paper, a four-channel microfluidic chip was designed. Its characteristics were as follows: it relied on laminar flow and diffusion mechanisms to establish and maintain a concentration gradient; it was suitable for observation of cell migration in different concentration gradient environment under a single microscope field; four cell isolation zones (20 μm width) were integrated into the microfluidic device to calibrate the initial cell position, which ensured the accuracy of the experimental results. In particular, we used COMSOL Multiphysics software to simulate the structure of the chip, which demonstrated the necessity of designing S-shaped microchannel and horizontal pressure balance channel to maintain concentration gradient. Finally, neutrophils were incubated with advanced glycation end products (AGEs, 0, 0.2, 0.5, 1.0 μmol·L -1), which were closely related to diabetes mellitus and its complications. The migration behavior of incubated neutrophils was studied in the 100 nmol·L -1 of chemokine (N-formylmethionyl-leucyl-phenyl-alanine) concentration gradient. The results prove the reliability and practicability of the microfluidic chip.

Participação dos neutrófilos durante a implantação embrionária no desenvolvimento da placenta em camundongos / Participation of neutrophils during embryonic implantation in the placental development in mice

A implantação do embrião na parede uterina é um processo complexo que consiste na interação do blastocisto com as células epiteliais do útero, e depende de diferentes tipos celulares do microambiente uterino. Embora a literatura mostre a participação de neutrófilos neste processo, os dados ainda são incipientes para proposição da função exata destas células nos períodos iniciais da gestação. Dados do nosso grupo de pesquisa mostraram que neutrófilos pró-angiogênicos induzem a tolerância gestacional, e que a depleção de neutrófilos durante as fases iniciais da gestação prejudica a implantação do blastocisto e a progressão da gestação. Com base nestes resultados, o presente estudo visou investigar se a depleção de neutrófilos na fase pré-receptiva da janela de implantação do blastocisto altera a morfologia placentária. Para tanto, foi utilizado o modelo de gestação alogênica, onde camundongos fêmeas C57BL/6, após cruzamento com machos Balb/C foram tratadas com anticorpo anti-Ly6G ou isotipo no dia 1,5 da gestação (24 horas após a detecção do plug vaginal) em dose suficiente para manter a depleção de neutrófilos circulantes por 48 horas (200µg/ 500µL; i.p). No final da gestação (dia 18,5), o sangue periférico foi coletado e, em seguida, os animais foram submetidos a laparotomia para retirada da placenta, a qual foi submetida à análise histológica. As análises dos leucócitos circulantes evidenciaram a efetividade do tratamento para depleção de neutrófilos periféricos. A análise histológica mostrou alterações significativas na morfologia da placenta nos animais tratados com anti-Ly6G. Foram detectadas a redução da zona juncional, de células trofoblásticas e de fatores angiogênicos, como fator de crescimento do endotélio vascular (VEGF), e das moléculas de adesão intracelular-1 (ICAM-1) e de plaqueta e endotélio (PECAM-1). Esses dados evidenciam a importância dos neutrófilos nos primeiros dias de gestação para o desenvolvimento da placenta

Blastocyst implantation is a complex process, consisting of the interaction between blastocyst and uterine epithelial cells. Also, it is well known that the implantation site resembles an inflammatory response, with a profusion of recruited immune cells into the endometrial stroma and lumen from the blood. The role of macrophages, natural killers, and dendritic cells have been extensively studied, however, the participation of neutrophils in this process remains unclear. Data from our research group showed that pro-angiogenic neutrophils induced gestation tolerance, also peripheral neutrophils depletion at the time of active placental development led to smaller embryo sizes and abnormal placentation in mice. In this context, the present study aimed to investigate whether pharmacological depletion of neutrophils in mice in the blastocyst implantation phase alters placental morphology. Therefore, C7/BL/6 female mice, after mating with Balb/C males, were treated with an anti-Ly6G antibody or isotype on day 1 of gestation (after detection of the vaginal plug) at a dose sufficient to maintain the depletion of circulating neutrophils for 48 hours (200 µg/500µL; i.p). At the end of the gestational day (day 18), peripheral blood was collected, and then the animals were submitted to laparotomy for the placenta removal and subsequent histological analysis. The analysis of circulating leukocytes from neutrophils depleted mice showed a reduction of peripheral neutrophils up to 48 hours after antibody injection. The histological analysis showed significant alterations in the placenta morphology of the animals treated with anti-Ly6G. The morphometric analyses showed a reduction in the size of neutrophils depleted placenta due to diminished junctional zone and reduction of trophoblast cells. Also, it was observed a reduction of vascular endothelial growth factors (VEGF), reduction of adhesion molecules intracell-1 (ICAM-1), and platelets and endothelium (PECAM-1) positive cells in the junctional zone. In conclusion, these data show the importance of neutrophils on the first days of pregnancy for the development of the placenta

Changes of heparin-binding protein in severe burn patients during shock stage and its effects on human umbilical vein endothelial cells and neutrophils / 中华烧伤杂志

Objective: To investigate the changes of heparin-binding protein (HBP) in severe burn patients during shock stage and its effects on human umbilical vein endothelial cells (HUVECs) and neutrophils in vitro. Methods: Prospective observational and experimental research methods were used. Twenty severe burn patients who met the inclusion criteria and were admitted to the Department of Burns and Plastic Surgery of Affiliated Suzhou Hospital of Nanjing Medical University from August to November 2020 were included in severe burn group (12 males and 8 females, aged 44.5 (31.0, 58.0) years). During the same period, 20 healthy volunteers with normal physical examination results in the unit's Physical Examination Center were recruited into healthy control group (13 males and 7 females, aged 39.5 (26.0, 53.0) years). Enzyme-linked immunosorbent assay (ELISA) method was used to detect the protein expression levels of HBP and tissue inhibitor of metalloproteinase 1 (TIMP-1) in plasma of patients within 48 hours after injury in severe burn group and in plasma of volunteers in healthy control group. The correlation between protein expression of HBP and that of TIMP-1 in the plasma in the two groups was analyzed by Pearson correlation analysis. The fourth passage of HUVECs in logarithmic growth phase were used for the experiment. The HUVECs were divided into normal control group with routine culture (the same treatment below) and recombinant HBP (rHBP)-treated 12 h group, rHBP-treated 24 h group, and rHBP-treated 48 h group with corresponding treatment according to the random number table (the same grouping method below), and the mRNA expression of TIMP-1 in cells was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. The HUVECs were divided into normal control group and rHBP-treated 48 h group with corresponding treatment, and the protein expression of TIMP-1 in the cells was detected by Western blotting. The HUVECs were divided into normal control group, rHBP alone group, aprotinin alone group, and rHBP+aprotinin group treated with the corresponding reagents (with the final molarity of rHBP being 200 nmol/L and the final concentration of aprotinin being 20 μg/mL, respectively), cultured for 48 h, and ELISA was used to detect the protein expression of TIMP-1 in the culture supernatant of cells. The neutrophils were isolated from the peripheral venous blood of the aforementioned 10 healthy volunteers by immunomagnetic bead sorting, and the cells were divided into normal control group, recombinant TIMP-1 (rTIMP-1) alone group, phorbol acetate (PMA) alone group, and rTIMP-1+PMA group treated with corresponding reagents (with the final concentration of rTIMP-1 being 500 ng/mL and the final molarity of PMA being 10 nmol/L, respectively). After being cultured for 1 h, the expression of CD63 protein in cells was detected by immunofluorescence method, the positive expression rate of CD63 protein in cells was detected by flow cytometry, and the protein expression levels of HBP and myeloperoxidase (MPO) in the culture supernatant of cells were detected by ELISA. The normal control group underwent the above-mentioned related tests at appropriate time points. The number of samples was 3 in each group of cell experiment. Data were statistically analyzed with chi-square test, Mann-Whitney U test, Kruskal-Wallis H test, and Tamhane's T2 test. Results: The protein expression levels of HBP and TIMP-1 in the plasma of patients in severe burn group were 404.9 (283.1, 653.2) and 262.1 (240.6, 317.4) ng/mL, respectively, which were both significantly higher than 61.6 (45.0, 68.9) and 81.0 (66.3, 90.0) ng/mL of volunteers in healthy control group (with Z values of -5.41 and -5.21, respectively, P<0.01). The correlation between the protein expression of HBP and that of TIMP-1 in the plasma of volunteers in healthy control group was not strong (P>0.05). The protein expression of HBP was significantly positively correlated with that of TIMP-1 in the plasma of patients in severe burn group (r=0.64, P<0.01). Compared with that in normal control group, the mRNA expression of TIMP-1 in HUVECs was significantly increased in rHBP-treated 12 h group, rHBP-treated 24 h group, and rHBP-treated 48 h group (with t values of -3.58, -2.25, and -1.26, respectively, P<0.05). Western blotting detection showed that compared with that in normal control group, the protein expression of TIMP-1 in HUVECs in rHBP-treated 48 h group was significantly enhanced. After 48 h of culture, compared with that in normal control group, the protein expression level of TIMP-1 in the culture supernatant of HUVECs in rHBP alone group was significantly increased (t=9.43, P<0.05), while the protein expression level of TIMP-1 in the culture supernatant of HUVECs didn't change significantly in aprotinin alone group or rHBP+aprotinin group (P>0.05); compared with that in rHBP alone group, the protein expression level of TIMP-1 in the culture supernatant of HUVECs in rHBP+aprotinin group was significantly decreased (t=4.76, P<0.01). After 1 h of culture, the trend of CD63 protein expression in neutrophils detected by immunofluorescence method and that by flow cytometry were consistent in each group. After 1 h of culture, compared with that in normal control group, the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells in rTIMP-1 alone group all had no significant changes (P>0.05), while the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells were all significantly increased in PMA alone group and rTIMP-1+PMA group (with t values of 2.41, 3.82, 5.73, 1.05, 4.16, and 1.08, respectively, P<0.05 or P<0.01); compared with that in PMA alone group, the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells in rTIMP-1+PMA group were all significantly decreased (with t values of 5.26, 2.83, and 1.26, respectively, P<0.05 or P<0.01). Conclusions: The expression level of HBP in the plasma of severe burn patients is increased during shock stage. HBP can induce HUVECs to secrete TIMP-1 in vitro, and TIMP-1 can reduce the expression of CD63 molecule in human neutrophils.

Re-understanding the physiological and pathophysiological roles of neutrophils / 中华烧伤杂志

Neutrophils have always been considered as a short-lived and homogeneous cell type in the innate immune system, which have limited pro-inflammatory or anti-inflammatory effects. However, in recent 10 years, the understanding of neutrophils has been undergoing some kind of revival as researches progressed. The researches on the heterogeneity of neutrophils and the mechanism of their interaction with other immune cells have promoted the researchers to re-understand the physiological and pathophysiological roles of neutrophils. In the following decades, with the development of single-cell sequencing technology, spatial transcriptome sequencing technology, and multi-omics combined sequencing technology, researchers will have a better understanding of the biological behaviors of neutrophils. This paper briefly reviews the biological behaviors of neutrophils and their roles in various diseases in recent years.

Relationship between preoperative inflammatory indexes and prognosis of patients with rectal cancer and establishment of prognostic nomogram prediction model / 中华肿瘤杂志

Objective: To compare the prognostic evaluation value of preoperative neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), and systemic immune-inflammation index (SII) in rectal cancer patients. Nomogram survival prediction model based on inflammatory markers was constructed. Methods: The clinical and survival data of 585 patients with rectal cancer who underwent radical resection in the First Affiliated Hospital of Xi'an Jiao tong University from January 2013 to December 2016 were retrospectively analyzed. The optimal cut-off values of NLR, PLR, LMR, and SII were determined by the receiver operating characteristic (ROC) curve. The relationship between different NLR, PLR, LMR and SII levels and the clinic pathological characteristics of the rectal cancer patients were compared. Cox proportional risk model was used for univariate and multivariate regression analysis. Nomogram prediction models of overall survival (OS) and disease-free survival (DFS) of patients with rectal cancer were established by the R Language software. The internal validation and accuracy of the nomograms were determined by the calculation of concordance index (C-index). Calibration curve was used to evaluate nomograms' efficiency. Results: The optimal cut-off values of preoperative NLR, PLR, LMR and SII of OS for rectal cancer patients were 2.44, 134.88, 4.70 and 354.18, respectively. There was statistically significant difference in tumor differentiation degree between the low NLR group and the high NLR group (P<0.05), and there were statistically significant differences in T stage, N stage, TNM stage, tumor differentiation degree and preoperative carcinoembryonic antigen (CEA) level between the low PLR group and the high PLR group (P<0.05). There was statistically significant difference in tumor differentiation degree between the low LMR group and the high LMR group (P<0.05), and there were statistically significant differences in T stage, N stage, TNM stage, tumor differentiation degree and preoperative CEA level between the low SII group and the high SII group (P<0.05). The multivariate Cox regression analysis showed that the age (HR=2.221, 95%CI: 1.526-3.231), TNM stage (Ⅲ grade: HR=4.425, 95%CI: 1.848-10.596), grade of differentiation (HR=1.630, 95%CI: 1.074-2.474), SII level (HR=2.949, 95%CI: 1.799-4.835), and postoperative chemoradiotherapy (HR=2.123, 95%CI: 1.506-2.992) were independent risk factors for the OS of patients with rectal cancer. The age (HR=2.107, 95%CI: 1.535-2.893), TNM stage (Ⅲ grade, HR=2.850, 95%CI: 1.430-5.680), grade of differentiation (HR=1.681, 95%CI: 1.150-2.457), SII level (HR=2.309, 95%CI: 1.546-3.447), and postoperative chemoradiotherapy (HR=1.837, 95%CI: 1.369-2.464) were independent risk factors of the DFS of patients with rectal cancer. According to the OS and DFS nomograms predict models of rectal cancer patients established by multivariate COX regression analysis, the C-index were 0.786 and 0.746, respectively. The calibration curve of the nomograms showed high consistence of predict and actual curves. Conclusions: Preoperative NLR, PLR, LMR and SII levels are all correlated with the prognosis of rectal cancer patients, and the SII level is an independent prognostic risk factor for patients with rectal cancer. Preoperative SII level can complement with the age, TNM stage, differentiation degree and postoperative adjuvant chemoradiotherapy to accurately predict the prognosis of rectal cancer patients, which can provide reference and help for clinical decision.

Association between the Neutrophil-to-lymphocyte Ratio and New-onset Subclinical Macrovascular and Microvascular Diseases in the Chinese Population / 生物医学与环境科学（英文）

OBJECTIVE@#The association between neutrophil-to-lymphocyte ratio (NLR) with subclinical macrovascular and microvascular diseases has been less investigated. We sought to examine the association between NLR and new-onset subclinical macrovascular and microvascular abnormalities in the Chinese population.@*METHODS@#From a community cohort, we included 6,430 adults aged ≥ 40 years without subclinical macrovascular and microvascular diseases at baseline. We measured subclinical macrovascular and microvascular abnormalities separately using the ankle-brachial index (ABI), brachial-ankle pulse wave velocity (baPWV), and albuminuria.@*RESULTS@#During a mean follow-up of 4.3 years, 110 participants developed incident abnormal ABI, 746 participants developed incident elevated baPWV, and 503 participants developed incident albuminuria. Poisson regression analysis indicated that NLR was significantly associated with an increased risk of new-onset abnormal ABI, elevated baPWV, and albuminuria. Compared to overweight/obese participants, we found a much stronger association between NLR and subclinical vascular abnormalities in participants with normal weight. Furthermore, we found an interaction between the NLR and body mass index (BMI) on the risk of new-onset abnormal ABI ( P for interaction: 0.01).@*CONCLUSION@#NLR was associated with subclinical macrovascular and microvascular diseases in the Chinese population. Furthermore, in participants with normal weight, the association between NLR and subclinical vascular abnormalities was much stronger.

Effects of hydroxyurea on cytotoxicity, inflammation and oxidative stress markers in neutrophils of patients with sickle cell anemia: dose-effect relationship

ABSTRACT Introduction: Although the efficacy of hydroxyurea (HU) in inhibiting erythrocyte sickling has been well demonstrated, the action of this drug on human neutrophils and the mechanism by which it improves the manifestations of the disease have not been studied thoroughly. We aimed to investigate the cell viability, along with inflammatory and oxidative markers in the neutrophils of sickle cell anemia (SCA) patients and the effects of HU therapy on these cells, by evaluating the dose-responsiveness. Methods: In the present study, 101 patients (45 men and 56 women, aged 18-69 years) with SCA were divided into groups according to the use or not of HU: the SS group (without HU treatment, n = 47) and the SSHU group (under HU treatment, n = 54). The SSHU group was further stratified into subgroups according to the daily dose of the drug that patients already used: SSHU - 0.5 g (n = 19); SSHU - 1 g (n = 26) and SSHU - 1.5-2 g (n = 9). A control group (AA) comprised 50 healthy individuals. Neutrophils isolated from whole blood were analyzed using Trypan Blue, monoiodotyrosine (MTT) and lactate dehydrogenase (LDH) toxicity assays. Myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activities and concentrations of interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-α) and malonaldehyde (MDA) were also measured. Results: Neutrophils from SCA patients showed membrane fragility and a significant decrease in cell viability when analyzed by Trypan Blue (p < 0.05), MTT (p < 0.001) and LDH (p = 0.011), compared to the AA group. Levels of inflammatory (MPO, TNF-α, and IL-10) and oxidative markers (SOD, GSH-Px, and MDA) were also altered (p < 0.05) in these cells, showing a significant difference in the SSHU-1g and SSHU - 1.5-2 g groups, compared to the SS group. Treatment with HU reverted the levels of all markers to concentrations similar to those in healthy individuals in a positive dose-effect relationship. Conclusion: The HU did not generate a cytotoxic effect on neutrophils in SCA patients, but it modulated their oxidative and inflammatory mechanisms, promoting cytoprotection with a positive dose-effect.