Clinical and laboratory differences between pediatric hospitalized patients with sickle cell disease infected or not by SARS-CoV-2 / Diferenças clínicas e laboratoriais entre pacientes pediátricos hospitalizados com doença falciforme infectados ou não por SARS-CoV-2

Abstract Objective: The aim of this study was to identify clinical and complete blood count differences between pediatric hospitalized patients with sickle cell disease infected or not by SARS-CoV-2 and compare the complete blood count of patients with sickle cell disease infected by SARS-CoV-2 before hospitalization and on admission. Methods: This study was a single-center prospective cohort. Data were collected from medical records of pediatric inpatients with sickle cell disease under 18 years old infected or not with SARS-CoV-2 from the first visit to the hospital until discharge and from the last medical appointment. All patients were tested for SARS-CoV-2 by the real-time reverse transcription polymerase chain reaction. Results: Among 57 pediatric patients with sickle cell disease hospitalized from March to November 2020 in a Brazilian academic hospital, 11 (19.3%) had a positive result for SARS-CoV-2. Patients infected by SARS-CoV-2 had a higher prevalence of comorbidities than the ones who were not infected (63.6 vs. 30.4%; p=0.046). During hospital stay, no clinical or complete blood count differences between groups were found. There was a decrease in eosinophil count on hospital admission in patients with sickle cell disease infected by SARS-CoV-2 (p=0.008). Conclusions: Pediatric hospitalized patients with sickle cell disease infected by SARS-CoV-2 had more comorbidities and had a decrease in eosinophil count between hospital admission and the last medical appointment.

RESUMO Objetivo: Identificar diferenças clínicas e laboratoriais entre pacientes pediátricos hospitalizados com doença falciforme infectados ou não por SARS-CoV-2 e comparar o hemograma completo de pacientes com doença falciforme infectados por SARS-CoV-2 antes da hospitalização e durante a admissão. Métodos: Coorte prospectiva unicêntrica, cujos dados foram coletados em prontuários de pacientes pediátricos internados com doença falciforme, menores de 18 anos, infectados ou não com SARS-CoV-2, desde a primeira visita ao hospital até a alta e desde a última consulta médica. Todos os pacientes foram testados para SARS-CoV-2 pela transcrição reversa seguida de reação em cadeia da polimerase em tempo real. Resultados: Dos 57 pacientes pediátricos com doença falciforme internados de março a novembro de 2020 em um hospital universitário brasileiro, 11 (19,3%) apresentaram resultado positivo para SARS-CoV-2. Pacientes infectados pelo SARS-CoV-2 apresentaram maior prevalência de comorbidades do que aqueles não infectados (63,6 vs. 30,4%; p=0,046). Durante a internação hospitalar, não foram encontradas diferenças clínicas ou laboratoriais entre os grupos. Houve diminuição da contagem de eosinófilos na admissão hospitalar em pacientes com doença falciforme infectados pelo SARS-CoV-2 (p=0,008). Conclusões: Pacientes pediátricos hospitalizados com doença falciforme infectados pelo SARS-CoV-2 apresentaram mais comorbidades e diminuição da contagem de eosinófilos entre a admissão hospitalar e a última consulta médica.

Construction and validation of an early predictive model for intraoperative massive transfusion of red blood cells in patients with Stanford type A aortic coarctation / 中国输血杂志

【Objective】 To analyze the risk factors for intraoperative massive red blood cell (RBC) transfusion in patients with Stanford type A aortic dissection (TAAD), in order to develop a risk-prediction model and validate its predictive effect. 【Methods】 The clinical data of 233 patients with TAAD admitted to our hospital from July 2018 to June 2021 (modeling set) were retrospectively analyzed. They were divided into routine transfusion group (n=128, RBC≤8 U) and massive transfusion group (n=105, RBC>8 U). Risk factors for intraoperative massive RBC transfusion in TAAD patients were analyzed by multivariate logistic regression and a risk prediction model was developed. Calibration curve and receiver operating characteristic (ROC) curve were used to assess the accuracy and discrimination of the model. In addition, 61 TAAD patients admitted to our hospital from July 2021 to May 2022 (validation set) were used for external validation. 【Results】 The rate of intraoperative massive RBC transfusion in 233 TAAD patients was 45.06% (95% CI: 38.59%-51.69%). Logistic analysis showed that women, age >50 years, preoperative Hb≤131.50 g/L, intraoperative bleeding >720 mL, and CPB time >155 min were independent risk factors for massive intraoperative RBC transfusion (P<0.05). The intraoperative risk prediction model formula for massive RBC infusion was: -4.427+ 0.925×gender+ 1.461×age+ 2.081×preoperative Hb+ 1.573×bleeding volume+ 2.823×CPB time. The area under the ROC curve of the modeling set and validation set were 0.904 (95% CI: 0.865-0.943) vs 0.868 (95%CI: 0.779-0.958), and the slopes of the calibration curves all converged to 1, indicating that the model predicted the risk of intraoperative massive RBC infusion in TAAD patients in good consistency with the actual risk of massive infusion. The decision curve shows that the model exhibits a positive net benefit with a threshold probability of 0.15-0.67 and has a high clinical application value. 【Conclusion】 The prediction model constructed based on the risk factors of intraoperative massive RBC infusion in TAAD patients can effectively predict the risk of intraoperative massive RBC infusion with high clinical predictive efficacy.

Preliminary analysis of qualitative changes in suspended red blood cells under different low-pressure and hypoxic conditions / 中国输血杂志

【Objective】 To evaluate the quality of suspended red blood cells (SRBC) under low-pressure and hypoxic conditions, in order to lay a theoretical foundation for airline delivery, air drop and storage of SRBC on plateau. 【Methods】 The low-pressure and hypoxic conditions (0.026 MPa and 0.047 MPa) were simulated by a pressure control device. SRBC were divided into 3 groups (5 bags/group, 1.5 U/bag). Each group was stored in 0.026 MPa environment at 2℃-6℃ for 24 hours (named 0.026 MPa), in 0.047MPa environment at 2℃-6℃ for 7 days (named 0.047 MPa), and in a blood storage refrigerator at 4℃ (named the control), respectively. The storage cell characteristics were examined on day 2, 9, 14, 28 and 35. 【Results】 We found that HCT, MCV, K+, Na+, FHb, hemolytic ratio, 2, 3-DPG and rheological properties in group 0.026 MPa and 0.047 MPa were not significant different compared with that in control (P>0.05). The consumptions of Glu in group 0.026 MPa and 0.047 MPa were significantly higher (P<0.05), and a transient increase in LAC concentration of group 0.026 MPa and 0.047MPa were observed, compared to the control. 【Conclusion】 The conditions of low-pressure and hypoxia have no significant effect on the quality of suspended red blood cells.

Recurrent neural network prediction on clinical usage of red blood cells / 中国输血杂志

【Objective】 To explore the prediction of clinical red blood cells (RBCs) consumption under major public health emergencies, so as to provide scientific basis for blood collection and blood inventory management. 【Methods】 The clinical consumption of different types of RBCs in Yichang from 2001 to 2017 was analyzed and modeled using the recurrent neural network (RNN) model, and the clinical RBCs consumption between January 2019 and December 2021(36 months) were scientifically predicted. 【Results】 The RNN model showed good prediction performance. The root mean square errors (RMSE) of RNN prediction values of A, B, O, AB type of RBCs were 156.7, 133.4, 204.2 and 51.3, respectively, with the average relative errors (MRE) at 6.4%, 6.6%, 8.5% and 7.1%, respectively. The model predicted the changing trend of RBCs consumption during the first round of COVID-19 outbreak (January to June, 2020) and forecasted the lowest level of consumption in February 2020 and a subsequent recovery in growth. The prediction of RBCs consumption during the second round of COVID-19 pandemic (January to June, 2021) was of high accuracy. For example, the relative errors of RNN models for A type RBCs consumption were 5.2% in Feb 2021 (the lowest level, 1 621.5 U) and 2.5% in May 2021 (the highest level, 2 397.0 U). 【Conclusion】 The artificial intelligence RNN model can predict clinical RBCs consumption well under major public health emergencies.

Discussion on the collection and supply mode of RhD negative blood suitable for prefecture-level city: a retrospective analysis / 中国输血杂志

【Objective】 To explore a RhD negative blood collection and supply mode suitable for Liaocheng area and improve the blood demand satisfaction rate of RhD negative patients. 【Methods】 Under different collection and supply modes (collection mode: in 2020, not advocate walk-in donation by RhD negative repeat blood donors, but in 2021 and 2022 were the opposite; supply mode: in 2020, type A, B, O and AB RBCs were frozen within 6 days of blood collection without inventory, but in 2021 and 2022, they were stored of 2-6 U and the remaining were frozen within 6 days of blood collection), RhD negative blood was divided into Type A, B, O and AB, appointment donation and walk-in donation, first donation and repeat donation, cold storage red blood cells(RBCs), frozen RBCs and frozen thawed deglycerolized RBCs, and the collection and supply data of each observation group from 2020 to 2022 were compared. Based on whether blood demand of RhD negative patients was met, the patients were divided into the group of going to other places for medical treatment, the group of RhD positive blood transfusion and the group of RhD negative blood transfusion to analyze the blood usage. 【Results】 From 2020 to 2022, the proportion of RhD negative blood donated by repeat appointment donors decreased year by year (P<0.05); the proportion of RhD negative blood donated by repeat walk-in donors increased year by year (P<0.05); the proportion of frozen thawed deglycerolized RBCs to RhD negative RBCs increased year by year (P<0.05); the proportion of cold storage RBCs distributed as RhD negative and RhD positive decreased year by year respectively (P<0.05); the proportion of the increase in frozen RBCs inventory to the current year's frozen RBCs inventory decreased year by year (P<0.05). The satisfaction rate of blood demand of RhD negative patients increased year by year (P<0.05), with the number of patients with RhD positive blood transfusion and going to other places for medical treatment decreased year by year respectively(P<0.05). 【Conclusion】 In Liaocheng area, it is appropriate to encourage RhD negative blood donors to donate blood randomly, and A, B, O and AB type cold storage RBCs kept in 2-6 U inventory respectively. When exceeding the inventory, frozen RBCs are prepared within 6 days, which can improve the blood demand satisfaction rate of RhD negative patients and avoid sending excessive RhD negative RBCs to clinical use as RhD positive RBCs.

Animalexperiment on regulating effect of stored red blood cells transfusion on BMDMs in inflammatory conditions / 中国输血杂志

【Objective】 To discuss the regulating effect of stored red blood cells (RBCs) transfusion on BMDMs in inflammatory conditions, and the relationship between stored RBCs transfusion and inflammatory response induced by bacterial infection. 【Methods】 Forty C57BL/6 male mice of 6-8 weeks (18-22 g/mouse) were randomly divided into experimental group and control group. Each mouse was infected with 200 µL Pseudomonas aeruginosa injecting into the tail vein, and 400 µL fresh (storage >14 d) and stored RBCs (storage 0.05). F4/80 of experimental group and control group 2, 4 and 8 hours after RBCs infused were 1.83±0.11 vs 0.75±0.06, 0.46±0.06 vs 0.33±0.06 (P0.05), respectively. iNOS, TNF-α, MCP1 of M1 in liver of experimental group and control group 2, 4 and 8 hours after RBCs infused were respectively: iNOS 3.44±0.20 vs 2.46±0.08, 9.25±0.55 vs 2.67±0.12, 2.80±0.08 vs 2.39 ±0.01; TNF-α 1.69±0.22 vs 1.13±0.03, 1.44±0.24 vs 0.96±0.09, 1.31±0.05 vs 0.96±0.06; MCP1 4.96±0.08 vs 4.28±0.27, 4.63±0.04 vs 2.07±0.09, 2.28±0.19 vs 1.33±0.03 (P0.05). 【Conclusion】 Stored RBCs infusion can greatly promote the M1 polarization of BMDMs in liver.

Exploration of the relationship between the storage time of leukodepleted red blood cell and transfusion adverse reactions / 中国输血杂志

【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Factors influencing hemolysis rate of leukocyte-reduced red blood cells at the end of preservation period in blood donors / 中国输血杂志

【Objective】 To compare the current standards and explore the influencing factors for hemolysis rate of leukocyte-reduced red blood cells at the end of the preservation period, in order to formulate reasonable internal control indicators. 【Methods】 A retrospective analysis was performed on hemolysis rate of 427 samples of leukocyte-reduced red blood cells at the end of the preservation period in Nanning Blood Center from 2015 to 2022. Compared with the current standard for hemolysis rate at the end of the preservation period (GB 18469-2012 Quality Requirements for Whole Blood and Component Blood), the differences were analyzed, and the factors influncing the hemolysis rate were analyzed in terms of different blood donor groups. 【Results】 1) Among the 427 samples, the hemolysis rate of 418 (97.89%) did not exceed 0.4%, all lower than 0.8%; 2)the hemolysis rate of the male group was higher than that of the female group; 3) the hemolysis rate of the 18-29 years old group was lower than that of the 30-39 year old group and the 40-60 year old group, with statistically significant difference; 4) in terms of occupation, the hemolysis rate of students was the lowest, and the differences between groups were statistically significant; 5) no statistical significance was found in ethnicity and blood type. 【Conclusion】 Statistics indicated that gender, age, blood donation volume and occupation of blood donors were the influencing factors of hemolysis rate. The current standard is obviously higher in the qualified range of blood quality control in Nanning. It is advisable to formulate a reasonable quality control strategy with internal control index of hemolysis rate set <0.4%, which is conducive to making accurate evaluation of internal quality control and ensuring blood safety.

Distribution and transfusion efficacy of unexpected antibody / 中国输血杂志

【Objective】 To investigate the clinical characteristics and antibody distribution as well as evaluate the transfusion efficacy in unexpected antibody positive patients. 【Methods】 A total of 12 235 patients from January 1, 2022 to March 31, 2023 who hospitalized in our hospital and applied for blood transfusion were selected, and those with unexpected antibody were included. The clinical data, including gender, age, diagnosis, blood type, history of transfusion and pregnancy were collected for antibody distribution analysis. Patients who received transfusion were grouped according to the DAT results and the components of red blood cells transfused, and the Hb values of each group before and after transfusion were compared. 【Results】 Among12 235 patients, 118 were positive for antibody screening, with a prevalence of 0.96%. The antibodies from Rh system were the most common (27.43%, 48/175), followed by MNS system (8.57%, 15/175) and Lewis system (6.29%, 11/175), mainly anti-E (18.29%, 32/175), anti-M (8.00%, 14/175) and anti-Lea (5.71%, 10/175). In addition, 62 transfused patients were divided into group A with suspended red blood cell transfusion and group B with washed red blood cell transfusion for positive DAT, and group C for negative DAT. Hb values (g/L) pre- and post-transfusion were 59.19±15.67 vs 77.52±15.09 in group A, 56.35±14.08 vs 74.44±15.63 in group B, 56.00±12.06 vs 75.00±4.73 in group C, respectively. The Hb values of post-transfusion for three groups were all higher than those of pre-transfusion (P<0.05). 【Conclusion】 Anti-E from Rh system is the most common antibody in patients with unexpected antibody. Appropriate red blood cells transfusion with Hb increases by an average of 6-7 g/L per 1 U of red blood cells indicating good transfusion efficacy. For positive DAT patients, transfusion of suspended red blood cell is feasible.

Research update on DNA methylation in peripheral blood cells as a risk factor for coronary heart disease / 中华检验医学杂志

Coronary heart disease (CHD) is a kind of cardiovascular diseases originated from atherosclerosis (AS), and chronic inflammation is one of the pathological characteristics. The peripheral blood leukocytes, especially mononuclear cells, play an important role in the AS processes. Recently, in a series of Epigenome-Wide Association Studies (EWAS), multiple DNA differential methylation sites in peripheral blood cells were found to be statistically associated with CHD, which suggested that these DNA differential methylation sites might serve as new risk factors for CHD. The recognition of the variant of DNA methylation as a common epigenetic nucleic acid modification in the occurrence and development of CHD, is ongoing. DNA methylation has the potential to become warning biomarkers, which might provide new ideas and evidences for mechanistic studies of CHD.

Hemoglobin S identification in blood donors: A cross section of prevalence

ABSTRACT Introduction: In Brazil, the sickle cell trait (SCT) has an average prevalence of 4% in the general population and 6-10% among Afro-descendants. Although SCT is highly prevalent, a large segment of the population ignores their status. The Therapeutic Guidelines prohibit the transfusion of SCT red blood cells into patients with hemoglobin disorders or severe acidosis and newborns. Methods: This was a cross-sectional study with data from 37,310 blood donation candidates. The study included only eligible first-time donors qualified to be tested for the presence of hemoglobin S (HbS) at the Fundação Hemominas Juiz de Fora, Brazil. The variables studied were gender, skin color, age, type of donation, place of birth, blood type, result of the solubility test for hemoglobin S (HbST) and hemoglobin electrophoresis (HbEF). Statistical analysis was performed using the Q square test and the Kappa index of agreement for comparing biochemical methods. This project was approved by the National Research Ethics Committee. Results: The analysis of first-time donor data showed that 7166 were considered eligible. A total of 127 of the 7166 donors were carriers of SCT (1.77%). Among the blood donors, 73.23% were from the local area. The HbST and HbEF were found to be 100% in concordance. Sensitivity was not tested in the present study. Conclusions: The HbST is highly specific for identifying the HbS, but sensitivity was not tested in this study. The screening of blood donors for abnormal hemoglobins is useful, helping to detect and counsel heterozygous people. The study seeks to identify the prevalence of SCT in a region of Brazil.

Asociación entre la variación de glóbulos blancos y las manifestaciones clínicas, anticuerpos e infecciones en adultos con lupus eritematoso sistémico / White blood cells count variation in systemic lupus erythematosus: its association to disease manifestations, antibodies and risk of infections

Introducción: la asociación de leucopenia, linfopenia y neutropenia con la presencia de autoanticuerpos, manifestaciones clínicas e infecciones en pacientes con lupus eritematoso sistémico (LES) no está bien establecida. Los objetivos de este estudio fueron analizar los cambios en los recuentos de leucocitos y linfocitos en pacientes con LES y su asociación con manifestaciones clínicas, autoanticuerpos y riesgo de infecciones. Materiales y métodos: se recolectaron retrospectivamente los valores de leucocitos, linfocitos y neutrófilos. Se agruparon a los pacientes en cinco categorías: recuento de glóbulos blancos normales, leucopenia (persistente o intermitente) y linfopenia (persistente o intermitente). Se registraron las manifestaciones clínicas, los autoanticuerpos acumulados, el daño, la mortalidad, las infecciones y los tratamientos inmunosupresores recibidos de cada paciente. Resultados: se incluyeron 89 pacientes. La linfopenia (89%) fue la anormalidad más frecuente. La leucopenia intermitente y la persistente se detectaron en el 44% y en el 11% de los pacientes, respectivamente. La linfopenia intermitente y la persistente se hallaron en el 44% y en el 45% de los casos. En el análisis univariado, la presencia de rash discoide se asoció a leucopenia (20,4 vs. 5,1; p=0,059) y el tratamiento con mofetil micofenolato a un recuento normal de leucocitos (p=0,046). El compromiso neurológico se asoció a recuento normal de linfocitos (22,2% vs. 0% y 7,5%; p=0,027); los pacientes con anti-RNP (anti ribonucleoproteína nuclear) presentaron más frecuentemente linfopenia persistente (47% vs. 15,4% y 20%; p=0,007). Ninguno de los grupos se asoció a una mayor prevalencia de infecciones. En el análisis multivariado, el mofetil micofenolato se asoció negativamente a leucopenia (OR 0.33 IC 95% 0,1-0,9; p=0,042) y el compromiso neurológico se asoció negativamente a linfopenia (OR 0.08; p=0,022). Conclusiones: en el análisis univariado, el rash discoide se asoció a leucopenia y el anti-RNP a linfopenia. Al ajustar por otras variables significativas, el tratamiento con mofetil micofenolato se asoció a un recuento normal de leucocitos, mientras que las manifestaciones neurológicas se relacionaron a linfocitos normales. No se demostró asociación de las infecciones con ninguno de los grupos.

Introduction: leukopenia, lymphopenia and neutropenia association to clinical manifestations and infections in systemic lupus erythematosus (SLE) is not well defined. The objectives were to analize leucocytes and lymphocytes variations in SLE patients and their association to clinical manifestations, autoantibodies and infections risk. Materials and methods: total white blood cell (WBC) count, lymphocyte, and neutrophils counts were collected retrospectively. Data were grouped into normal WBC cell count, persistent or intermittent leucopenia and lymphopenia. Disease manifestations, accumulated autoantibodies, damage, mortality, infections and immunosuppressants ever received were registered. Results: study sample included 89 patients. Lymphopenia (89%) was the most common abnormality. Intermittent and persistent leukopenia were detected in 44% and 11% cases. Intermittent and persistent lymphopenia were found in 44% and 45% cases. In univariate analysis, discoid rash was associated to leukopenia (20.4 vs 5.1 p=0.059) and mycophenolate treatment to normal leukocyte count (p=0.046). Patients with neurological disorder tended to have normal lymphocyte counts rather than intermittent or persistent lymphopenia (22.2% vs 0% and 7.5% p=0.027); patients with anti-RNP tended to belong to the persistent lymphopenia group (47% vs 15.4% and 20% p=0.007). Infections were not associated to any of the categories. In multivariate analysis mycophenolate was negatively associated to leukopenia (OR 0.33 95% CI 0.1-0.9 p=0.042) while neurological disorder was negatively associated to lymphopenia (OR 0.08 p=0.022). Conclusions: in univariate analysis, discoid rash was associated to leukopenia and anti-RNP to lymphopenia. When adjusted to other significant variables, mycophenolate was related to normal leukocyte while neurological manifestations were to normal lymphocyte counts. Infections were not associated to any of the categories.

BLOOD CELL MORPHOLOGY AND LEUKOCYTE DIFFERENTIAL OF MORELET'S CROCODILE (Crocodylus moreletii) (CROCODYLIDAE) / Morfología de células sanguíneas y recuento leucocitario del cocodrilo de Morelet (Crocodylus moreletii) (CROCODYLIDAE)

ABSTRACT The objective of the study was to determine the morphological characteristics of peripheral blood cells (erythrocytes, leukocytes, thrombocytes) and the leukocyte differential count (heterophils, eosinophils, basophils, lymphocytes, monocytes, azurophils) of captive Morelet's crocodiles (Crocodylus moreletii) from Veracruz, Mexico. Peripheral blood from 80 apparently healthy farmed crocodiles (39 subadults [19 females, 20 males] and 41 adults [18 females, 23 males]) was examined for morphology through stained blood smears and manual count was used for the leukocyte differential. Blood was collected during the non-breeding (n = 42) and breeding (n = 38) seasons. Blood examination indicated similar morphological characteristics of blood cells in subadult and adult individuals and in females and males in both seasons. Erythrocytes were the largest blood cells and lymphocytes the smallest. The leukocyte differential count showed that lymphocytes were the most abundant leukocytes and basophils the least numerous. The percentages of some leukocytes showed difference by season (non-breeding and breeding) in subadult and adult males (p < 0.05) and by size (subadults and adults) in males and females but only in the non-breeding season (p < 0.05). The leukocytes that showed the greatest variation were lymphocytes, heterophils and eosinophils. The knowledge of blood cell morphology and the leukocyte differential count in healthy farmed Morelet's crocodiles will allow the accurate diagnosis of some diseases of captive and wild individuals.

RESUMEN El objetivo del estudio fue determinar las características morfológicas de células sanguíneas periféricas (eritrocitos, leucocitos, trombocitos) y el conteo diferencial de leucocitos (heterófilos, eosinófilos, basófilos, linfocitos, monocitos, azurófilos) del cocodrilo de Morelet (Crocodylus moreletii) mantenido en cautiverio en Veracruz, México. Se examinó sangre periférica de 80 cocodrilos aparentemente sanos criados en granja (39 subadultos [19 hembras, 20 machos] y 41 adultos [18 hembras, 23 machos]) mediante frotis sanguíneos teñidos para determinar la morfología celular y mediante conteo manual para el diferencial de leucocitos. La sangre se recolectó durante las temporadas no reproductiva (n = 42) y reproductiva (n = 38). El examen sanguíneo indicó similar morfología de células sanguíneas en individuos subadultos y adultos, así como en hembras y machos, en ambas temporadas. Los eritrocitos fueron las células sanguíneas más grandes y los linfocitos las más pequeñas. El conteo diferencial de leucocitos mostró que los linfocitos fueron los leucocitos más abundantes y los basófilos los menos numerosos. Los porcentajes de algunos leucocitos mostraron diferencia por época (no reproductiva y reproductiva) en machos subadultos y adultos (p < 0,05) y por talla (subadultos y adultos) en machos y hembras sólo en temporada no reproductiva (p < 0,05). Los leucocitos que tuvieron la mayor variación fueron linfocitos, heterófilos y eosinófilos. El conocer la morfología de las células sanguíneas y el conteo diferencial de leucocitos en individuos sanos de cocodrilo de Morelet criados en granja permitirá diagnosticar con precisión algunas enfermedades de individuos manejados en cautiverio y también de individuos silvestres.

Spectrum of Reactive Morphological Alterations in Leucocytes in HIV Patients.

Background: The earliest hematological manifestation of HIV infection can occur at the time of primary infection. Several peripheral blood abnormalities are known in HIV. However, not many studies have reported morphological changes in white blood cells in antiviral naïve patients and even fewer have compared them with HIV seropositive patients. Material and methods: Peripheral blood smears from 100 patients were studied. 50 of them were newly diagnosed HIV seropositive cases and 50 were HIV seronegative. A peripheral blood smear was made to study morphology of various white blood cells A written informed consent was taken. Statistical analysis of the data obtained was done. Results: Morphological abnormalities in the lymphocytes had been the most significant finding, which included nuclear lobulation, nuclear convolution and plasmacytoid and monocytoid features. Neutrophils also showed various morphological abnormalities. Most common were hypogranulation, hypersegmented nuclei and cytoplasmic vacuolations in descending order of frequency. Monocytes also revealed few morphological abnormalities including nuclear irregularities and cytoplasmic vacuolations. Conclusion: The observations from this study will serve as a guide to clinicians as treatment of these patients should include supplements to monitor and improve these changes.

Influence of low-dose occupational radiation exposure on peripheral blood cells in radiation workers / 国际生物医学工程杂志

Objective:To investigate the effect of low-dose ionizing radiation on blood cell parameters of radiation workers.Methods:A total of 124 staff members engaged in radiology were selected into the observation group, and they were divided into 4 subgroups of physicians, physicists, technicians, and maintainer according to their jobs. A total of 130 non-radiation-related staff members from the same hospital were selected into the control group. Blood cell parameters of peripheral blood of all subjects from 2016 to 2019 were collected, and the differences in blood cell parameters between the radiation group and the control group as well as 4 subgroups of the control group were analyzed and compared, and the correlation between the differences in blood cell parameters and the cumulative radiation dose was compared.Results:Compared with the control group, the white blood cell count, neutrophil count, red blood cell count and hemoglobin count in the observation group were lower than those in the control group (all P<0.05). There are no significant differences in cumulative radiation dose among different types of work (all P>0.05). Correlation analysis showed that the blood cell parameters of peripheral blood cells were not significantly correlated with the cumulative radiation dose. The blood cell count changes after 4-year low-dose ionizing radiation between the physicist group, the technician group and the maintainer sub-group were significantly different (all P<0.05), but the above differences were not related to the cumulative radiation dose (all P>0.05). Conclusions:Under the same exposure and protection conditions, the blood cell counts of different radiation-related workers are not significantly different, and the long-term cumulative radiation dose has no significant correlation with blood cell parameters. Therefore, peripheral blood cell parameters can no longer be used as a good indicator to reflect radiation damage, and it is urgent to find more convenient, intuitive and sensitive indicators of radiation damage.

Blood cells predict the outcomes of patients with acute ischemic stroke after endovascular therapy / 国际脑血管病杂志

Endovascular therapy has been proved to be the best treatment for acute ischemic stroke caused by large vessel occlusion, but its efficacy is closely associated with time, so it is of great significance to obtain biomarkers that can predict its outcome early. Blood cells can reflect inflammatory reaction, which is very low cost and easy to obtain. This article expounds the predictive value of blood cells on the outcomes of acute ischemic stroke after endovascular therapy.

Comparación de la eficacia de la transfusión de paquetes globulares de ≤ 15 vs ≥ 16 a 40 días de extracción sobre los efectos hemodinámicos y gasométricos de pacientes con traumatismo craneoencefálico severo en la Unidad de Cuidados Intensivos / Comparison of the efficacy of the transfusion of globular packages of ≤ 15 vs ≥ 16 to 40 days of extraction on the hemodynamic and gasometric effects of patients with severe traumatic brain in the Intensive Care Unit / Comparação da eficácia da transfusão de hemácias de ≤ 15 vs ≥ 16 a 40 dias de extração sobre os efeitos hemodinâmicos e gasométricos de pacientes com traumatismo cranioencefálico grave na Unidade de Terapia Intensiva

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Abstract: Introduction: since the beginning of blood banks, refinements in laboratory processes have allowed longer red blood cell storage times. While advantageous to the logistics of stock management, the clinical impact of RBC storage duration prior to transfusion remains uncertain and a topic of growing interest. Although the structural, biochemical, and impermeable changes that red blood cells undergo during storage are well described, the evidence that this storage injury translates into adverse clinical outcomes for patients receiving blood with longer storage times remains controversial. Objective: to compare the efficacy of the transfusion of globular packages of ≤ 15 days of extraction vs. globular packages ≥ 16-40 days of extraction in the hemodynamic and gasometric repercussion of patients with severe traumatic brain injury (TBI), hospitalized in the Intensive Care Unit of HE CMNO. Material and methods: a controlled, single-center, randomized, single-blind, prospective and comparative clinical trial was conducted, where patients between 18 and 80 years of age with severe postoperative cranioencephalic trauma who required blood transfusions were invited to participate. Participants were included in two random groups, group F and group E. Group F was administered packed cells ≤ 15 days of extraction. Group E will be administered globular packages ≥ 16-40 days of extraction. During the study, various hemodynamic and biochemical variables were measured before, during, and after blood transfusion, and a comparison of the results obtained between groups F and E was subsequently made. Results: in the present study, a total of 26 patients with severe TBI were included and who were transfused with red blood cell packs, of the total number of patients, 13 were transfused with a «standard¼ red blood cell pack and another 13 patients with a «fresh¼ red blood cell pack. The average days of transfusion after extraction were 18 and 14 days for the «standard¼ and «fresh¼ packs, respectively (p ≤ 0.001). Twenty-one infectious events were reported, 11 in the group of patients who were transfused with the standard pack and 10 in the fresh pack group. Mortality at 28 days was estimated in 31% of the patients transfused with the standard pack and in 23% of the patients with the fresh pack (RR, for 28-day mortality of 0.90 [95% CI 0.56-144]). The median duration time in the intensive care unit was 8 days for both groups (0.32 SD), and of the days associated with the ventilator, 15 days were observed for the group of patients with the standard package and 7 for the group with fresh package (0.60 SD), without discovering statistically significant differences in these variables. However, in this analysis statistically significant differences were found for the gasometric parameters of central venous oxygen saturation, cardiac output (Fick) and lactate before and after transfusion in favor of the group of patients transfused with fresh pack (p ≤ 0.05). Conclusion: the results of this study infer that there is no association between the storage time of transfused red blood cells and the presence of adverse clinical outcomes with longer storage times. In both groups, transfusions were equally safe and effective. The researchers refer to the sample size as a limitation for this study.

Resumo: Introdução: desde o surgimento dos bancos de sangue, o refinamento dos processos laboratoriais permitiu tempos de estocagem cada vez mais longos para as hemácias. Embora vantajoso para a logística do gerenciamento de estoque, o impacto clínico da duração do armazenamento de glóbulos vermelhos antes da transfusão permanece incerto e um tópico de interesse crescente. Embora as alterações estruturais, bioquímicas e imunológicas pelas quais as hemácias sofrem durante o armazenamento sejam bem descritas, as evidências de que essa lesão de armazenamento resulta em resultados clínicos adversos para pacientes que recebem sangue armazenado por mais tempo permanecem controversas. Objetivo: comparar a eficácia da transfusão de concentrado de hemácias ≤ 15 dias de extração vs concentrado de hemácias ≥ 16-40 dias de extração nas repercussão hemodinâmica e gasométrica dos pacientes com traumatismo cranioencefálico (TCE) grave, internados em Unidade de Terapia Intensiva do HE CMN Occidente. Material e métodos: realizou-se um ensaio clínico, controlado, unicêntrico, randomizado, simples-cego, prospectivo e comparativo, onde foram convidados a participar pacientes com idade entre 18 e 80 anos com traumatismo cranioencefálico grave submetidos a hemicraniectomia descompressiva com necessidade de transfusão sanguínea. Os participantes foram incluídos aleatoriamente em 2 grupos, grupo F e grupo E. Grupo F recebeu concentrado de hemácias ≤ 15 dias após a extração. O grupo E receberá concentrado de hemácias ≥ 16-40 dias após a extração. Durante o estudo, várias variáveis hemodinâmicas e bioquímicas foram medidas antes, durante e após a transfusão de sangue, e posteriormente foi feita uma comparação dos resultados obtidos entre os grupos F e E. Resultados: no presente estudo incluíram-se um total de 26 pacientes com TCE grave e que foram transfundidos com concentrado de hemácias, do total de pacientes 13 foram transfundidos com um concentrado de hemácias «padrão¼ e outros 13 pacientes com um concentrado de hemácias «fresco¼. A média de dias de transfusão após a extração foi de 18 e 14 dias para as pacotes «padrão¼ e «fresco¼, respectivamente (p ≤ 0.001). Relatou-se 21 eventos infecciosos, 11 no grupo de pacientes que receberam transfusão com concentrado padrão e 10 no grupo com concentrado fresco. A mortalidade em 28 dias foi estimada em 31% dos pacientes transfundidos com concentrado padrão e em 23% dos pacientes com concentrado fresco (RR, para mortalidade em 28 dias de 0.90 (IC 95% 0.56-144). A duração mediana na unidade de terapia intensiva foi de 8 dias para ambos os grupos (0.32 DP), sendo que 2 dias associados ao ventilador, foram observados 15 dias para o grupo de pacientes com concentrado padrão e 7 para o grupo de concentrado fresco (0.60 DE), sem descobrir diferenças estatisticamente significativas nas referidas variáveis.No entanto, nesta análise foram encontradas diferenças estatisticamente significativas para os parâmetros gasométricos de saturação venosa central de oxigênio, débito cardíaco (Fick) e lactato antes e após a transfusão em favor do grupo de pacientes transfundidos com concentrado fresco (p ≤ 0.05). Conclusão: os resultados deste estudo inferem que não há associação entre o tempo de estocagem das hemácias transfundidas e a presença de desfechos clínicos adversos com tempos de estocagem mais longos. Em ambos os grupos, as transfusões foram igualmente seguras e eficazes. Os pesquisadores referem-se ao tamanho da amostra como limitante para este estudo.

The comparison of methods of the microscopic examination with Sternheimer-Malbin stain and UF-5000 analyzer for urine sediment / Эрүүл мэндийн лаборатори

Background@#Chronic kidney disease (CKD) is a global health problem. In Mongolia, urine is analyzed by methods of urine chemistry and urine sediment to diagnose kidney disease. The currently automated urine sediment analyzers have been widely used in clinical laboratories and are replacing traditional manual microscopic examination. Nonetheless, visual microscopic examination is still required in many cases. When chemical and sediment analyzers are used together, urine sediment could be confirmed under a microscope, if the results are inconsistent. Sternheimer-Malbin stain has contained a variety of dyes that help to distinguish particles (white blood cells, red blood cells, epithelial cells, casts, crystals, fatty drops, bacteria, yeast, trichomonas) in urine sediment, improve the differentiation between cell nuclei and cytoplasm, and provide more information about cell shape and image. </br> Therefore, the low-cost method that can be used on a daily basis.Although there are more than 4,500 laboratories in Mongolia that need to perform urinalysis, which is an important part of clinical laboratories, less than 10 percent of hospitals have fully automated sediment analyzers. For this reason, one of the most important issues in the clinical laboratories, the search for low-cost and useful methods for the analysis of urine sediments in order to provide access to services to the public. Our aim was the comparison of methods of the microscopic examination with Shternheimer-Malbin stain and fully automated UF-5000 analyzer for urine sediment. @*Methods@#There was a comparative study, people who served the Clinical Central Laboratory of Mongolia-Japan Hospital received permission to participate in this research. One hundred five fresh, first morning, clean catch mid-stream urine samples were collected in accordance with standard operating instructions for urinalysis, between November 2020 and May 2021. Sternheimer-Malbin (SM) staining and direct microscopy observation methods with Fuchs-Rosenthal counting chamber were used to red blood cells (RBC), white blood cells (WBC) and epithelial cells (EC) in urine samples. The agreements between the automated urine analyzer and microscopic methods were calculated using Cohen’s kappa (k) with 95% confidence intervals (CI).@*Results@#A total of 105 samples were collected and analysed in this study. The average age was 46.97±15.0and gender by 18% (n=19)were male and 82% (n=86) were female. </br> Compared to traditional manual methods and automated analyzer, the agreement within the same grade was 99/105 (94.3%) for erythrocytes, 96/105 (91.4%) for leukocytes, 92/105 (87.6%) for epithelial cells. And compared to Sternheimer-Malbin staining microscopy observation and automated analyzer, the agreement within the same grade was 98/105 (93.3%) for erythrocytes, 99/105 (94.3%) for leukocytes, 96/105 (91.4%) for epithelial cells. Agreement between traditional manual method and automated analyzer was higher than 85% and between Sternheimer-Malbin staining microscopy observation and automated analyzer was higher than 90%. The concordance between traditional manual method and automated analyzer was substantial (k=0.74, p<0.001; k=0.79, p<0.001) for RBC and EC, almost perfect (k=0.92, p<0.001) for WBC. Whereas the concordance between SternheimerMalbin staining microscopy observation and automated analyzer was substantial (k=0.70, p<0.001) for RBC, almost perfect (k=0.94, p<0.001; k=0.89, p<0.001) for WBC and EC. Comparison of Sysmex UF-5000 with microscopic particle counting methods resulted specificity was 98.9/100% for RBC, sensitivity was 97.7/95.3% and negative predictive value was 98.4/96.8% for WBC, sensitivity was 87.5/68.8% and negative predictive value was 97.8/94.7% for EC.@*Conclusion@#The Cohen’s k analysis result of comparisons between the SternheimerMalbin staining microscopic method and automated urine sediment analyzer showed significant almost perfect agreement (k=0.70-0.94, p<0.001). </br> The sensitivity and negative predictive value were high for both of WBC and EC were determined by Sternheimer-Malbin (SM) staining microscopy observation method. Results indicate the ability of a test to correctly identify those with the true positive and individual with a negative test result is truly negative better than comparison of Sysmex UF-5000 with traditional manual microscopic method assessment.

Cyclic arginine-glycine-aspartic acid-modified red blood cells for drug delivery:Synthesis and in vitro evaluation / 药物分析学报

Red blood cells(RBCs)are an excellent choice for cell preparation research because of their biocom-patibility,high drug loading,and long half-life.In this study,doxorubicin(DOX)was encapsulated with RBCs as the carrier.The biotin-avidin system binding principle was used to modify biotinylated cyclic arginine-glycine-aspartic acid(cRGD)onto RBC surfaces for accurate targeting,high drug loading,and sustained drug release.The RBC drug delivery system(DDS)was characterized,and the concentration of surface sulfur in the energy spectrum was 6.330％.The physical and chemical properties of RBC DDS were as follows:drug content,0.857 mg/mL;particle size,3339 nm;potential value,-12.5 mV;and cumu-lative release rate,81.35％.There was no significant change in RBC morphology for up to seven days.The results of the targeting and cytotoxicity studies of RBC DDS showed that many RBCs covered the surfaces of U251 cells,and the fluorescence intensity was higher than that of MCF-7 cells.The IC50 value of un-modified drug-loaded RBCs was 2.5 times higher than that of targeted modified drug-loaded RBCs,indicating that the targeting of cancer cells produced satisfactory inhibition.This study confirms that the RBC DDS has the characteristics of accurate targeting,high drug loading,and slow drug release,which increases its likelihood of becoming a clinical cancer treatment in the future.

How phosphorylation of RBCs band 3 protein affects storage quality of suspended red blood cells prepared from Tibetan high Hb population / 中国输血杂志

【Objective】 To investigate the quality changes of suspended red blood cells (SRBCs) prepared from the blood of Tibetan high Hb population, and explore the availability and safety of blood collected from Tibetan high Hb population. 【Methods】 The voluntary blood donors were grouped according to the Hb concentration at the initial screening: female blood donors from Tibet Autonomous Region (>3 500 m) with Hb≥190 g/L and male blood donors with Hb≥210 g/L were classified as plateau high hemoglobin group. A total of 13 male blood donors from Tibet Autonomous Region were recruited. And the female blood donors (n=13) with Hb(115~165) g/L and male blood donors (n=12) with Hb(120~185) g/L from Chengdu were classified as control group. Whole blood of 200 mL specification was centrifuged to remove the plasma, and MAP additive solution was added to prepare SRBCs, then SRBCs were divided into four aliquots (50 mL/bag and stored at 4℃. Parameters as blood routine, free Hb and hemolysis rate were measured aseptically at day 1, 14, 21, 35 of storage. And 10 mL SRBCs was used to extract membrane proteins for tyrosine phosphorylation detection of band 3 protein. 【Results】 The RBCs counts(×1012/L), hematocrit(%) and hemoglobin(g/L) of Tibetan high Hb group and control group were 6.76±0.95 vs 4.65±0.52, 63.3±6.8 vs 43.1±4.4 and 214.4±19.8 vs 143.2±16.9 (P<0.01). The erythrocyte deformability test on the day 1, 14, 21, 35 of storage showed that the deformability of SRBCs prepared from Tibetan high Hb group was significantly lower than that of the control group under shear stress of 3, 5.33, 9.49, 16.87, and 30 Pa, while the hemolysis rate of SRBCs prepared from the Tibetan high Hb group and the control group on the day 1, 14, 21, 35 were 0.050 2±0.040 2 vs 0.022 2±0.011 1, 0.055 4±0.043 vs 0.032 1±0.028 7, 0.061 2±0.025 9 vs 0.034 3±0.031 7 and 0.069 6±0.032 0 vs 0.044 0±0.033 3 (P<0.05). Western blotting showed that the cytoplasmic N-terminal Y21 of band 3 protein of SRBCs prepared from Tibetan high Hb group was highly phosphorylated. 【Conclusion】 The deformability of SRBCs prepared from the Tibetan high Hb group was significantly lower while the hemolysis rate of SRBCs was higher than that of the control group. The hemolysis rate of the SRBCs at the end of storage prepared from the Tibetan high Hb group meets the requirements of the national standard GB18469-2012(<0.8%). The increase of hemolysis rate of SRBCs prepared from the Tibetan high Hb group was closely related to the phosphorylation of band 3 protein.