Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation / Efeitos de oxidação e osmotolerância dos eritrócitos de Salvator merianae (Squamata: Teiidae) durante a hibernação

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or "tegu" lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou "teiú". Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or tegu lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou teiú. Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Hemoglobina como predictor del recuento de hematocrito y hematíes según edad y sexo en una población de Villa El Salvador en Lima-Perú

Objetivo: Evaluar el grado de correlación de la hemoglobina como posible predictor del hematócrito y recuento de hematíes según la edad y el sexo en una población del distrito de Villa El Salvador, Lima-Perú. Materiales y métodos: Estudio observacional, analítico, correlacional y prospectivo realizado a 550 hombres y 700 mujeres de todas las edades que acudieron a las campañas de prevención promocionales de salud mensuales, así como a las consultas médicas de rutina, desde enero de 2021 a junio de 2022. El muestreo fue no probabilístico por conveniencia. Las variables fueron sexo, grupo etario, hematocrito y recuento de hematíes; se aplicó la prueba de correlación de Spearman debido a que la prueba de normalidad de Kolmogorov-Smirnov mostró que la distribución no era normal. Se consideró un valor de p significativo menor del 0,05 con un intervalo de confianza al 95 %. Resultados: Hubo mayor prevalencia de anemia en el grupo de mujeres mayores de 11 años (35,10 %), mientras que los niveles de hemoglobina en menores de 11 años fueron, principalmente, normales en la mayoría de los pacientes. Se encontraron correlaciones altas y positivas entre hemoglobina y hematocrito cercanas a 1 en todos los grupos de edad y para ambos sexos. También se halló correlaciones moderadas y positivas entre infantes y niñas (Rho = 0,525), adolescentes y mujeres (Rho = 624), así como infantes y niños (Rho = 0,597). Conclusiones: Los niveles de hemoglobina se correlacionan de forma alta y positiva con el hematocrito. Es posible que el análisis simultáneo de hemoglobina y hematocrito sea clínica y económicamente innecesario en el cribado y en las campañas de prevención promocionales sobre anemia en grandes grupos poblacionales. Asimismo, la hemoglobina se correlaciona de forma moderada y positiva con los hematíes en niños de ambos sexos y adolescentes mujeres. Es necesario ampliar la investigación con estudios que profundicen las razones que causan que la correlación varíe en estos grupos.

Objective: To assess the degree of hemoglobin correlation as a possible predictor of hematocrit and red blood cell count according to age and sex in a population of the district of Villa El Salvador, Lima, Peru. Materials and methods: An observational, analytical, correlational and prospective study carried out with 550 men and 700 women of all ages who attended monthly preventive-promotional health campaigns, as well as routine medical consultations, from January 2021 to June 2022. Collected data consisted of hematocrit, red blood cells and hemoglobin counts. The Spearman's correlation coefficient was used. Results: There was a higher prevalence of anemia in the group of women older than 11 years (35.10 %), while hemoglobin levels in most children younger than 11 years were mainly normal. High and positive correlations between hemoglobin and hematocrit close to one were found in all age groups and both sexes. In addition, moderate and positive correlations were found in female infants and girls (Rho = 0.525), female adolescents (Rho = 624), as well as male infants and boys (Rho = 0.597). Conclusions: Hemoglobin levels are highly and positively correlated with hematocrit. Simultaneous hemoglobin and hematocrit testing may be clinically and economically unnecessary in screening and preventive-promotional campaigns on anemia conducted in large population groups. Likewise, hemoglobin correlates moderately and positively with red blood cell in children of both sexes and female adolescents. Further research is needed to deepen the reasons why correlation varies in these groups.

Ministernotomy in Aortic Root and Arch Surgery: Early Outcomes

ABSTRACT Introduction: Minimally invasive methods have become more preferred in cardiac surgery today. In this study, the comparative results of patients who underwent an aortic root, arch or hemiarch replacement by ministernotomy and full sternotomy in our clinic are presented. Methods: Between January 2017 and October 2019, a series of operations including aortic root, ascending aorta, and aortic arch replacements were performed on 278 patients. The ministernotomy technique was used in 25 of them. Twenty patients who underwent full sternotomy were selected and matched to this group for comparison. Results: The ministernotomy group had a longer cross-clamping time (128.3±30.8 vs. 104.7±23.4 min, P=0.007) but the total operating time was similar in the two groups (249.76±28.56 vs. 248.25±37.53 min, P=0.879). The number of red blood cell (RBC) transfusions per patient was higher in the full sternotomy group (4.65±3.74 vs. 2.44±1.85 unit, P=0.020). The ministernotomy group had shorter ventilation times (7.60±4.88 vs. 32.30±32.25 h, P<0.001) and shorter ICU stay (1.56±0.58 vs. 3.35±1.46 d, P<0.001). The 30-day mortality was 0% in the ministernotomy group. Conclusion: Early results of our study show that, in combined or isolated aortic root, ascending aorta, and aortic arch surgeries, ministernotomy can be applied with relatively safety and low mortality and morbidity rates.

Microfluidics-based study on the damage pattern of red blood cells and VWF / 国际生物医学工程杂志

Objective:To investigate the effects of shear stress magnitude and exposure time on the damage of blood component erythrocytes and von willebrand factor (VWF) based on microfluidic technology.Methods:A blood shear platform was built based on a microfluidic chip, samples were prepared under different shear stress magnitudes and exposure time lengths, free hemoglobin assay experiments were performed on blood samples, the hemolysis indices of different samples were measured, and the relative molecular masses of different samples of VWF were analyzed by immunoblotting and chemiluminescence imaging.Results:The quantitative relationships between the hemolysis index and the degradation rate of high relative molecular mass VWF with shear stress and exposure time followed the power function model well.Conclusions:The microfluidic experimental platform has the advantages of a precise and controllable internal microenvironment and easy and rapid detection, which can be used for the quantitative study of blood damage patterns.

Red blood cell distribution width and white matter hyperintensities / 国际脑血管病杂志

Dementia is one of the age-related diseases with the fastest increase in incidence. With the increasingly severe aging problem of the population, it further exacerbates the demand and burden of the healthcare system. Red blood cell distribution width (RDW) is an easily obtainable blood routine indicator that reflects the variability of red blood cell size. As an early marker of dementia risk, researches have shown an association between white matter hyperintensities (WMHs) and RDW. This article reviews the relationship between RDW and WMHs.

Validation of Blood Transfusion Risk Scores (TRACK and TRUST) in a Cardiac Surgery Service in Brazil

ABSTRACT Introduction: Transfusion of red blood cells is recurrent in cardiac surgery despite the well-established deleterious effects. Identifying patients with higher chances of requiring blood transfusion is essential to apply strategic preventive measures to reduce such chances, considering the restricted availability of this product. The most used risk scores to predict blood transfusion are the Transfusion Risk and Clinical Knowledge (TRACK) and Transfusion Risk Understanding Scoring Tool (TRUST). However, these scores were not validated for the Brazilian population. The objective of this study was to assess the accuracy of TRACK and TRUST scores in estimating the need for postoperative transfusion of red blood cell concentrates (TRBCC) after cardiac surgery. Methods: A clinical retrospective study was conducted using the database of a Brazilian reference service composed of patients operated between November 2019 and September 2021. Scores were compared using Mann-Whitney U test. Hosmer-Lemeshow goodness of fit test assessed calibration of the scores. Accuracy was assessed using the area under the receiver operating characteristic curve (AUC). All analyses considered a level of significance of 5%. The study was approved by the research ethics committee (CAAE 55577421.4.0000.5201). Results: This study assessed 498 patients. Only the TRACK score presented good calibration (P=0.238; TRUST P=0.034). AUC of TRACK was 0.678 (95% confidence interval 0.63 to 0.73; P<0.001), showing a significant accuracy. Conclusion: Between the scores analyzed, only the TRACK score showed a good calibration, but low accuracy, to predict postoperative TRBCC after cardiac surgery.

Investigation of the relationship between red blood cell distribution width and mortality in patients with hemophagocytic lymphohistiocytosis: a retrospective study

ABSTRACT BACKGROUND: Red blood cell distribution width (RDW) is related to sepsis-related mortality. Hemophagocytic lymphohistiocytosis (HLH) is a syndrome caused by severe infection, tumors, or autoimmunity without a specific diagnosis. OBJECTIVE: To explore the correlation between RDW and mortality in patients with HLH. DESIGN AND SETTING: A retrospective study conducted in a hospital in China. METHODS: A total of 101 inpatients with HLH from January 1, 2017 to December 31, 2021 were divided into non-survivor (n = 52) and survivor (n = 49) groups. A non-parametric test was used to analyze demographic, clinical, and laboratory data between groups. Independent variables with P < 0.05 were analyzed using binary logistic regression to screen out mortality-related variables. Selected variables were subjected to multivariate logistic regression analysis, and those with strong correlations were screened. Receiver operating characteristic (ROC) curves of strongly correlated variables and area under curve (AUC) values were obtained. RESULTS: The APACHE II score, RDW, and platelet (PLT) and fibrinogen (FIB) levels (P < 0.05) different significantly. RDW, PLT, FIB were correlated with mortality. The AUC values of RDW, PLT, and FIB were 0.857, 0.797, and 0.726, respectively. RDW was associated with mortality in patients with HLH (P < 0.01, cut-off value: 16.9). The sensitivity and specificity of predicting mortality were 97.96% and 96.1%, respectively. CONCLUSION: Logistic regression analysis showed a correlation between RDW and patients' mortality. Therefore, RDW can be used to predict mortality in patients with HLH.

Research progress on stable gene transfection methods of Plasmodium falciparum / 中国热带医学

@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

Effect of phenotypes of Duffy blood group on chemokine storage and chemokine scavenging function of erythrocytes / 中国输血杂志

【Objective】 To investigate the effect of phenotypes of Duffy blood group on chemokine storage and chemokine scavenging function of erythrocytes. 【Methods】 Twenty-four erythrocyte samples were collected and tested Duffy blood phenotype using the anti-human globulin method, and erythrocyte CCL2, CCL5, CXCL8, and CCL11 content and their chemokine scavenging function using ELISA. The expression of Duffy antigens on erythrocytes was detected using a flow analyzer. 【Results】 The difference in CCL2 content(41.1±14.7 pg/mL vs 63.1±20.8 pg/mL)of erythrocyte lysate between Fy(a+b-) and Fy(a+b+) phenotype was statistically significant (P0.05).The difference in the scavenging function of CCL2(1471±202.1 pg/mL vs 1860±267.5 pg/mL)and CCL5 (848.5±461.7 pg/mL vs 1797±546.1pg/mL) between Fy(a+b-) and Fy(a+b+) phenotype were statistically significant (P0.05).The difference in Duffy antigen expression (mean fluorescent intensity:105.3±20.45 vs 111.9±18.30)on erythrocytes between Fy(a+b-) and Fy(a+b+) phenotype was not statistically significant (P>0.05). 【Conclusion】 The Fy(a+b+) and Fy(a+b-) phenotypes of the Duffy blood group can affect the chemokine storage and scavenging function of erythrocytes. Fy(a+b+) phenotypes are able to store more chemokines and have a stronger chemokine scavenging function than Fy(a+b-) phenotypes.

Role of caspase-3 in aggravation of oxidative stress injury in stored red blood cells by CPB: cell experiment / 中华麻醉学杂志

Objective:To evaluate the role of caspase-3 in aggravation of the oxidative stress injury in stored red blood cells (sRBCs) by cardiopulmonary bypass (CPB).Methods:Eight patients with type O blood undergoing valve replacement with CPB were selected, blood samples 7 ml were collected through the central venous catheter before CPB, blood samples 20 ml were collected at 2 h after CPB, and the plasma before and after CPB was obtained after centrifugation. Eight samples of sRBCs 14 ml of blood type O stored for 7-14 days were collected and each sample was divided into A, B, C and D groups with 3.5 ml in each group. Normal saline 30 μl was added to group A and group B, 10% dimethyl sulfoxide 30 μl was added to group C, and 3.5 mmol/L Z-DEVD-fmk solution 30 μl was added to group D. The sRBCs were pretreated in a 37 ℃ water bath for 2 h in the four groups. Then group A was incubated with plasma before CPB, group B, C and D were incubated with plasma at 2 h of CPB, and four groups were incubated for 48 h in a thermostatic oscillator at 37 ℃ and 80 rpm. At 2, 24 and 48 h of incubation, the activity of caspase-3 and concentration of ATP were determined by enzyme-linked immunosorbent assay, the concentrations of glutathione (GSH) and free hemoglobin (FHb) were measured by colorimetry, and the exposure rate of cell membrane phosphatidylserine (PS) was detected by flow cytometry.Results:With the prolongation of incubation time, the activity of caspase-3, exposure rate of PS at cell membrane and concentration of FHb were gradually increased, and the concentrations of ATP and GSH were gradually decreased in the four groups ( P<0.05). Compared with group A, the activity of caspase-3 was significantly increased at each time point of incubation in group B and group C, the activity of caspase-3 was increased at 24 and 48 h of incubation in group D, and the concentration of ATP was decreased at 24 and 48 h of incubation, and the concentration of GSH was decreased and the concentration of FHb was increased at each time point of incubation in group B, group C and group D ( P<0.05). Compared with group B and group C, the activity of caspase-3 was significantly decreased, the concentrations of ATP and GSH were increased, and the exposure rate of PS at cell membrane and concentration of FHb were decreased at 24 and 48 h of incubation in group D ( P<0.05). There was no significant difference in the parameters mentioned above between group B and group C ( P>0.05). Conclusions:Caspase-3 is involved in aggravation of oxidative stress injury in sRBCs by CPB.

Mechanism of epidural scar hyperplasia after laminectomy induced by blood siltation / 中华骨科杂志

Objective:To explore the mechanism of epidural scar tissue hyperplasia induced by erythrocyte rupture and release of interleukin-33 (IL-33) after laminectomy in mice.Methods:In the zoological experiment, the operation group (Laminectomy) and the sham operation group were set, and HE staining and Masson staining were performed to test for blood accumulation in the operation area after laminectomy in mice. Then 12 wild-type mice with 6-8 week old were selected and divided into 4 groups: the sham operation group, the operation group (normal saline control), the pure red blood cell intervention operation group, the whole blood intervention operation group. The normal saline (100 mg/kg) was injected into the postoperative area. The red blood cells or whole blood with the same volume were injected into the postoperative area in the pure red blood cell intervention group and the whole blood intervention group. The postoperative recovery of mice in each group was observed. The levels of fibronectin in the postoperative scar tissues of mice in four groups were detected by western blot technology, and the degree of postoperative epidural scar hyperplasia was directly observed by immunohistochemistry. In the cytological experiment, the wild-type mouse erythrocyte normal saline group, the control group of IL-33 knockout mouse erythrocyte normal saline, the wild-type mouse erythrocyte lysis group, and the IL-33 knockout mouse erythrocyte lysis group were set. The levels of IL-33 in the red blood cells of four groups were detected by western blot. Then, a blank wild-type mouse erythrocyte control group, a wild-type mouse relative to the control group (only secondary antibody added to test for non-specific binding), a wild-type mouse erythrocyte group and an IL-33 knockout mouse erythrocyte group (to test for antigen specificity of the primary antibody) were set. Immunofluorescence staining was performed on the erythrocytes of four groups and the level of IL-33 was detected by flow cytometry.Results:HE staining and Masson staining after laminectomy showed that there was blood stasis in the local incision area of mice in the operation group. The epidural scar hyperplasia in the incision area of mice after whole blood or red blood cells intervention was higher, especially in the whole blood intervention group. IL-33 expression was almost undetectable in the wild-type erythrocyte normal saline control group, the IL-33-knockout erythrocyte normal saline control group, and the IL-33-knockout erythrocyte lysis group, while significant IL-33 expression was detectable in the wild-type erythrocyte lysis group. Immunofluorescence staining showed that IL-33 was expressed in and on the erythrocyte membrane of wild-type mice, while non-specific expression of IL-33 or a very small amount of IL-33 was almost undetectable in the other three groups. The immunofluorescence intensities of IL-33 in the four groups were 0.62±0.41, 60.17±4.39, 16.78±7.43 and 0.61±0.03, respectively ( F=281.90, P<0.001). The expression of IL-33 in the erythrocyte group of wild-type mice was the highest ( P<0.05). According to the results of flow cytometry, except for the trace amount of IL-33 detected in the wild-type mouse erythrocyte group, the expression of IL-33 in the other three groups was basically 0. The ratios of fibronectin to β-actin in the modeling area of the four groups gradually increased, and the ratios were 0.79±0.09, 1.26±0.23, 1.79±0.05 and 2.29±0.58, respectively, and the differences were statistically significant ( F=12.86, P=0.002). Fibronectin in the operation area of the three operation groups (normal saline control group, red blood cell intervention group and whole blood intervention group) was significantly higher than that of the sham operation group. The immunohistochemical staining results of fibronectin in the modeling area of the four groups were the same as those in western blot experiment. The average optical density values of fibronectin in each group were 0.09±0.01, 0.18±0.01, 0.22±0.01 and 0.24±0.01, respectively, and the difference was statistically significant ( F= 210.7, P<0.001). Conclusion:There is indeed blood accumulation in the surgical area after laminectomy in mice, and it can aggravate the hyperplasia of epidural scar tissue. Erythrocyte is the main component in blood, and there is a large amount of IL-33 expression in the inner and outer membrane of erythrocyte membrane. The mechanism of promoting the proliferation of epidural scar tissue may be related to the release of IL-33 by erythrocyte lysis.

Diagnostic and prognostic values of the ratio of red blood cell distribution width to platelet count for hepatitis B and liver cirrhosis / 中国基层医药

Objective:To analyze the diagnostic and prognostic values of the red blood cell distribution width-to-platelet count ratio (RPR) for hepatitis B and liver cirrhosis.Methods:The clinical data of 80 patients with hepatitis B and liver cirrhosis who were diagnosed and treated in Yiwu Central Hospital from June 2020 to August 2021 were retrospectively analyzed. These patients were included in the hepatitis B and liver cirrhosis group. They were subdivided into survival ( n = 69) and death ( n = 11) groups according to their prognosis outcomes. Eighty patients with chronic hepatitis B were included in the chronic hepatitis B group. Eighty healthy controls who concurrently underwent physical examination were included in the control group. The diagnostic and prognostic values of RPR, aspartate aminotransferase-to-platelet ratio index (APRI), and fibrosis index based on four factors (FIB-4) for hepatitis B and liver cirrhosis were analyzed. Results:Red blood cell distribution width, alanine transaminase, and aspartate transaminase in the hepatitis B and liver cirrhosis group and chronic hepatitis B group were significantly higher compared with the control group (all P < 0.05). Platelet count in the hepatitis B and liver cirrhosis group and chronic hepatitis B group was significantly lower than that in the control group (both P < 0.05). Red blood cell distribution width in the hepatitis B and liver cirrhosis group was significantly higher than that in the chronic hepatitis B group [(18.25 ± 3.28)% vs. (14.67 ± 2.15)%, t = 8.16, P < 0.05]. Platelet count, alanine transaminase, and aspartate transaminase levels in the hepatitis B and liver cirrhosis group were (78.47 ± 11.43) × 10 9/L, (49.48 ± 6.85) U/L, (45.86 ± 6.28) U/L, respectively, which were significantly lower than (133.36 ± 18.42) × 10 9/L, (128.36 ± 15.40) U/L, (98.67 ± 14.41) U/L in the chronic hepatitis B group ( t = -22.65, -41.86, -30.05, all P < 0.05). PRP, APRI, and FIB-4 in the hepatitis B and liver cirrhosis group were (0.23 ± 0.05), (1.85 ± 0.44), (4.25 ± 0.81) respectively, which were significantly higher than (0.11 ± 0.02), (1.46 ± 0.33), (3.38 ± 0.63) in the chronic hepatitis B group ( t = 19.93, 6.34, 7.58, all P < 0.001). The RPR, APRI, and FIB-4 in the death group were (0.25 ± 0.08), (1.97 ± 0.48), (4.52 ± 1.31), respectively, which were significantly higher than (0.18 ± 0.05), (1.68 ± 0.40), (3.69 ± 1.21) in the survival group ( t = 3.94, 2.17, 2.09, all P < 0.05). The receiver operating characteristic curve revealed that PRP has an extremely high value in diagnosing hepatitis B and liver cirrhosis and predicting the death of patients with hepatitis B and liver cirrhosis. Conclusion:RPR has an extremely high value in diagnosing hepatitis B and liver cirrhosis and predicting the prognosis of this disease.

Eritrocitos humanos como biomodelo farmacológico de citoprotección antioxidante para la evaluación de nuevas alternativas terapéuticas en pacientes con COVID-19 / Human erythrocytes as pharmacological biomodel of antioxidant cytoprotection for the evaluation of new therapeutic alternatives in patients with COVID-19

Introducción: El estrés oxidativo puede afectar las membranas biológicas de diferentes tipos celulares en el organismo, lo cual se ha evidenciado en los daños a los tejidos y órganos de pacientes con COVID-19, por lo cual las investigaciones recientes están relacionadas con la búsqueda de fármacos citoprotectores y antioxidantes que minimicen estos daños. Objetivo: Evaluar los eritrocitos humanos como biomodelo farmacológico de citoprotección antioxidante. Métodos: Se evaluó el modelo de citotoxicidad en eritrocitos inducido por peróxido de hidrógeno y se valoró el sistema de diagnóstico propuesto en un ensayo de citoprotección en eritrocitos, con el empleo del ácido ascórbico como sustancia de referencia. Resultados: Para la concentración de eritrocitos utilizada se logró un modelo de citotoxicidad a la concentración de 10 mM de peróxido a los 30 minutos de incubación. La sustancia de referencia empleada no mostró signos de citotoxicidad en el test de hemólisis. En el ensayo de citoprotección se evidenció un efecto farmacológico del referente, con un valor del índice de citoprotección de 12,71 µg/mL. El estudio de microscopía óptica mostró daños morfológicos severos en los eritrocitos tratados con peróxido de tipo esferocitos, equinocitos y esferoequinocitos, que disminuyeron significativamente en presencia de dicha sustancia de referencia. Conclusiones: El biomodelo farmacológico propuesto puede ser empleado en la evaluación de nuevas alternativas terapéuticas con propiedades citoprotectoras antioxidantes para el tratamiento de pacientes con COVID-19.

Introduction: The oxidative stress can affect the biological membranes of different cellular types in the organism, which has been evidenced in the damages to the tissues and organs of patients with COVID-19, reason why the recent investigations are related to the search of cytoprotector and antioxidant drugs that minimize these damages. Objective: To evaluate the human erythrocytes as pharmacological biomodel of antioxidant cytoprotection. Methods: The cytotoxicity pattern was evaluated in erythrocytes induced by peroxide of hydrogen and the system of diagnosis proposed was valued in a cytoprotection assay in erythrocytes, with the use of ascorbic acid as reference substance. Results: For the concentration of erythrocytes used a cytotoxicity model was achieved to the concentration of 10 mM of peroxide at 30 minutes of incubation. The substance of reference used didn't show cytotoxicity signs in the hemolysis test. In the cytoprotection assay a pharmacological effect of the referent was evidenced, with a value of the cytoprotection index of 12.71 µg/mL. The study of optic microscopy showed severe morphological damages in the erythrocytes treated with peroxide of spherocytes, echinocytes and spheroechinocytes type that significantly diminished in presence of this reference substance. Conclusions: The proposed pharmacological biomodel can be used in the evaluation of new therapeutic alternatives with antioxidant cytoprotector properties for the treatment of patients with COVID-19.

Macrophages: key players in erythrocyte turnover

ABSTRACT The development of red blood cells (RBCs), or erythropoiesis, occurs in specialized niches in the bone marrow, called erythroblastic islands, composed of a central macrophage surrounded by erythroblasts at different stages of differentiation. Upon anemia or hypoxemia, erythropoiesis extends to extramedullary sites, mainly spleen and liver, a process known as stress erythropoiesis, leading to the expansion of erythroid progenitors, iron recruitment and increased production of reticulocytes and mature RBCs. Macrophages are key cells in both homeostatic and stress erythropoiesis, providing conditions for erythroid cells to survive, proliferate and differentiate. During RBCs aging and injury, macrophages play a fundamental role again, performing the clearance of these cells and recycling iron for new erythroblasts in development. Thus, macrophages are crucial components of the RBCs turnover and in this review, we aimed to cover the main known mechanisms involved in the process of birth and death of RBCs, highlighting the importance of macrophage functions in the whole RBC lifecycle.

A Retrospective Study of the Relationship Between Blood Transfusion and 30-Day Postoperative Outcomes in Patients Undergoing Isolated Off-Pump Coronary Artery Bypass Grafting

ABSTRACT Introduction: The objective of this single-center study it to retrospectively analyze the relationship between transfusion and 30-day postoperative outcomes in patients undergoing isolated off-pump coronary artery bypass grafting. Methods: Perioperative data of 2,178 patients who underwent isolated off-pump coronary artery bypass grafting from 2018 to 2019 were collected. A 1:1 propensity score matching was performed to control for potential biases between patients who received blood transfusion and those who did not. After propensity score matching, we analyzed the clinical outcomes of transfusion and non-transfusion patients. Postoperative complications and the survival of patients within 30 days after surgery in both groups were analyzed. Kaplan-Meier survival curve and log-rank test were used for survival analysis. Results: The total blood transfusion rate of all patients was 29%, including red blood cell (27.6%), plasma (7.3%), and platelet (1.9%). Four hundred and forty patients in each group were compared after propensity score matching. There were no significant differences in the incidence of stroke, myocardial infarction, atrial fibrillation, acute kidney function injury, and sternal wound infection of both groups (P>0.05). However, higher incidence of postoperative pulmonary infection and more mechanical ventilation time and days of stay in the intensive care unit and postoperative in-hospital stay were associated with blood transfusion (P<0.05). The 30-day cumulative survival rate of the transfusion group was lower than that of the control group (P<0.05). Conclusion: Perioperative blood transfusion increases the risks of postoperative pulmonary infection and short-term mortality in off-pump coronary artery bypass grafting patients.

Avaliação da citotoxicidade em hemácias de humanos do extrato etanólico de Praxelis clematidea (Griseb.)

Introdução: Espécies da família Asteraceae são conhecidas por apresentarem propriedades aromática, cosmética e terapêutica; tendo diversas pesquisas que evidenciou potencial medicinal dessa família. Dentre as espécies de Asteraceae, está Praxelis clematidea, que é rica em substâncias químicas como flavonoides, terpenóides e esteroides, as quais podem desempenhar uma série de atividades biológicas. Objetivo: Verificar o potencial tóxico do extrato etanólico das folhas de P. clematidea frente à células sanguíneas humanas, afim de determinar a toxicidade teórica dessa espécie. Métodos: Para a realização do teste de atividade citotóxica foram preparadas suspensões sanguíneas dos tipos A, B e O, que posteriormente foram misturadas a concentrações distintas do extrato etanólico por 1 (uma) hora. A hemólise foi quantificada por espectrofotometria em comprimento de onda de 540 nm. Resultados: O extrato etanólico das folhas de P. clematidea em diferentes concentrações apresentou baixa citoxicidade contra os eritrócitos humanos in vitro, enfatizando o produto como uma possível opção viável para a indústria de medicamentos fitoterápicos. No entanto, é importante elucidar que mais estudos in vivo precisam ser realizados para confirmar esse perfil toxicológico do extrato.

SUMMARY Introduction: Species of the Asteraceae family are known to have aromatic, cosmetic and therapeutic properties; having several researches that evidenced medicinal potential of this family. Among the species of Asteraceae, there is Praxelis clematidea, which is rich in chemical substances such as flavonoids, terpenoids and steroids, which can perform a series of biological activities. Aim: To verify the toxic potential of the ethanolic extract of P. clematidea leaves against human blood cells, in order to determine the theoretical toxicity of this species. Method: For the performance of the cytotoxic activity test, blood suspensions of types A, B and O were prepared, which were subsequently mixed at different concentrations of the ethanolic extract for 1 (one) hour. Hemolysis was quantified by spectrophotometry at a wavelength of 540 nm. Results: The ethanolic extract of P. clematidea leaves in different concentrations showed low cytotoxicity against human erythrocytes in vitro, emphasizing the product as a possible viable option for the herbal medicine industry. However, it is important to clarify that more in vivo studies need to be carried out to confirm this toxicological profile of the extract.

Introducción: Se sabe que las especies de la familia Asteraceae tienen propiedades aromáticas, cosméticas y terapéuticas; habiendo varias investigaciones que evidenciaron el potencial medicinal de esta familia. Entre las especies de Asteraceae, se encuentra Praxelis clematidea, que es rica en sustancias químicas como flavonoides, terpenoides y esteroides, que pueden realizar una serie de actividades biológicas. Objetivo: Verificar el potencial tóxico del extracto etanólico de hojas de P. clematidea contra las células sanguíneas humanas, con el fin de determinar la toxicidad teórica de esta especie. Metodos: Para realizar la prueba de actividad citotóxica se prepararon suspensiones de sangre de los tipos A, B y O, que luego se mezclaron a diferentes concentraciones del extracto etanólico durante 1 (una) hora. La hemólisis se cuantificó mediante espectrofotometría a una longitud de onda de 540 nm. Resultados: El extracto etanólico de hojas de P. clematidea a diferentes concentraciones mostró baja citotoxicidad contra eritrocitos humanos in vitro, destacando el producto como una posible opción viable para la industria de la fitoterapia. Sin embargo, es importante aclarar que es necesario realizar más estudios in vivo para confirmar este perfil toxicológico del extracto.

Development of a cost-effective ex vivo lung perfusion system for lung transplantation in India

Background & objectives: Standard donor lung preservation with cold flush and storage allows up to six hours between retrieval of lungs from the donor and transplantation in the recipient. Ex vivo lung perfusion (EVLP) systems mimic physiological ventilation and perfusion in the donor lungs with potential for prolonged lung preservation and donor lung reconditioning. In this study, it was aimed to perform EVLP on discarded donor lungs using a locally developed EVLP system. Methods: Equipment that are routinely used for cardiac surgeries were collected and a functional EVLP system was assembled. This system was used on five pairs of lungs retrieved from brain-dead organ donors. The lungs were ventilated and pulmonary circulation was continuously perfused with a solution containing oxygen and nutrients for four hours. The system was tested without red blood cells (RBCs) added to the solution (acellular group; n=3; A1, A2 and A3) and also with RBCs added to the solution (cellular group; n=2; C1 and C2). Results: The EVLP system was successfully used in four (A1, A2, A3 and C2) of the five lung pairs. Mechanical and gas exchange functions of the lungs were preserved in these lung pairs. One lung pair (C1) worsened and developed pulmonary oedema. Histopathological examination of all five lung pairs was satisfactory at the end of the procedure. Major challenges faced were leakage of solution from the system and obstruction to drainage of RBCs containing solution from the lungs. Interpretation & conclusions: The results of the present study suggest that, it is possible to maintain the lungs retrieved for transplantation in a physiological condition using a locally prepared EVLP system and a solution without RBCs.

Diagnosis and treatment of 50 children with mycoplasma pneumoniae pneumonia with pseudo-macrocytic erythrocytes / 中国小儿急救医学

Objective:To investigate the clinical characteristics, diagnosis and treatment of mycoplasma pneumoniae(MP) pneumonia in children with pseudo-macrocytic erythrocytes.Methods:The clinical data of 50 children with mycoplasma pneumoniae pneumonia with pseudo-macrocytic erythrocytes in the Department of Pediatrics at Shengjing Hospital of China Medical University from January 2019 to August 2020 were analyzed retrospectively.Results:Among the 50 cases, there were 32 boys and 18 girls.The blood routine examination showed that pseudo-macrocytic erythrocytes, red blood cells decreased significantly, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration increased significantly, as well as MP-IgM was positive.Sixteen cases were complicated with herpes simplex virus infection, one with Epstein Barr virus infection, and six with both herpes simplex virus and Epstein Barr virus infection at the same time.All 50 cases were MP pneumonia, pulmonary imaging showed lobar pneumonia, and 25 cases were complicated with pleural effusion, including 32 cases of refractory MP pneumonia.The clinical symptoms of three cases were extrapulmonary manifestations, hemolytic anemia and diagnosed with cold agglutinin syndrome.In 36 children with D-dimer more than 252 μg/L, one case had femoral vein thrombosis and one case had pulmonary embolism.Conclusion:Pseudo-macrocytic phenomenon may play important roles in clinical etiological diagnosis, severity of disease and refractory MP pneumonia.The children with hemolytic anemia suggest cold agglutinin syndrome, and the hypercoagulable state of MP infection may be related to the aggregation of red blood cells caused by cold agglutinin in MP infection.

Erythrocyte sphingolipid species as biomarkers of Alzheimer's disease / 药物分析学报

Diagnosing Alzheimer's disease(AD)in the early stage is challenging.Informative biomarkers can be of great value for population-based screening.Metabolomics studies have been used to find potential biomarkers,but commonly used tissue sources can be difficult to obtain.The objective of this study was to determine the potential utility of erythrocyte metabolite profiles in screening for AD.Unlike some commonly-used sources such as cerebrospinal fluid and brain tissue,erythrocytes are plentiful and easily accessed.Moreover,erythrocytes are metabolically active,a feature that distinguishes this sample source from other bodily fluids like plasma and urine.In this preliminary pilot study,the erythrocyte metab-olomes of 10 histopathologically confirmed AD patients and 10 patients without AD(control(CTRL))were compared.Whole blood was collected post-mortem and erythrocytes were analyzed using ultra-performance liquid chromatography tandem mass spectrometry.Over 750 metabolites were identified in AD and CTRL erythrocytes.Seven were increased in AD while 24 were decreased(P＜0.05).The ma-jority of the metabolites increased in AD were associated with amino acid metabolism and all of the decreased metabolites were associated with lipid metabolism.Prominent among the potential bio-markers were 10 sphingolipid or sphingolipid-related species that were consistently decreased in AD patients.Sphingolipids have been previously implicated in AD and other neurological conditions.Furthermore,previous studies have shown that erythrocyte sphingolipid concentrations vary widely in normal,healthy adults.Together,these observations suggest that certain erythrocyte lipid phenotypes could be markers of risk for development of AD.