Fragmented Red Blood Cells Research Articles

A-153 Study of the evaluation of fragment flag and percentage schistocyte count in MINDRAY® BC-6800 PLUS in a public pediatric hospital in Argentina

Abstract Background Schistocytes are fragmented red blood cells (FRCs) that arise from the mechanical destruction of red blood cells (RBCs). They indicate the possible presence of a thrombotic microangiopathy (TMA). TMA includes hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura(TTP). ICSH defined schistocytes as 4 well stablish shapes always smaller than intact RBCs. The gold standard for the identification of schistocytes is microscopic evaluation. A schistocyte count should be considered clinically meaningful if it represent the main morphological RBCs abnormality in the periferic blood smear (PBS). The results should be expressed as a percentage, after counting at least 1000 RBCs in optimal areas of the PBS. The cut off for TMA was previously stablished in above 1% (adult population) and a higher percentaje in pre term newborns (above 5%). Some blood cell count analyzers have a flag for FRC, and some of them also provide methods for an automated FRC count, through the measurement of forward scatter and the intensity of fluorescence or 2-dimensional optical analysis. The Mindray® BC 6800 PLUS provides a flag for fragments and the research FRC% parameter that quantifies fragmented RBCs. When the flag and the FRC% are combined, can bring value to diagnostic TMA. Objetives Evaluate the sensitivity and specificity of the fragment flag to predict schistocyte on the PBS of pediatric samples. Evaluate the utility of FRC% as TMA diagnostic tool and compare with the gold standard. Methods 725 pediatric samples were analyzed (0-18 years).16 of them was patient samples diagnosed with TMA. All blood cell counts were determined using Mindray®BC-6800 PLUS. Cut off 1% was considered for FRC% in TMA. Inclusion criteria for TMA samples include thrombocytopenia, increased lactate dehydrogenase and decreased hemoglobin concentration. Schistocyte counting was carried out from May-Grunwald-Giemsa-stained PBS according to the recommendations proposed by ICSH. Only ICSH established shapes were kept as valid. Results Regarding the sensitivity of the fragment flag, it was 84.62% with a specificity of 91.13% to detect the presence of schistocytes in PBS samples. In the Bland-Altman analysis, the FRC% shows a difference with a tendency to underestimation with respect to the observation on the PBS, which was less at lower percentages of schistocytes. The sensitivity of the FRC% to TMA was 75% while the specificity was 94.36% and a negative predictive value (NPV) of 99.41%. Conclusions Fragments flag represents a potentially powerful tool for screening of squistocytes in pediatric samples. Although sensibility and NPV for FRC% authomatical counts were acceptable, a few false-negative samples were found. Presence of FRC% below a threshold of 1% could exclude TMA.However PBS microscopic observation is still the gold standard to determine squistocytes presence. For the purpose of complete our study and to get further larger scale prospective data, we started collecting all data for all samples entering the laboratory in order to improve an authomatical method that should have a more rapid turn-around time at lower costs.

Veno-venous extracorporeal membrane oxygenation in managing acute respiratory distress syndrome associated with hemolytic uremic syndrome and septic shock: a case report.

Veno-venous extracorporeal membrane oxygenation (VV-ECMO) is a rescue therapy for severe respiratory failure in which conventional mechanical ventilation therapy is unsuccessful. Hemolysis during VV-ECMO support arises from multiple factors associated with organ damage and poor outcomes. Therefore, close and prompt monitoring is needed. Hemolytic uremic syndrome (HUS) is characterized by hemolysis, acute renal failure, and thrombocytopenia. Hemolytic features of the disease may complicate VV-ECMO management. A 26-year-old man with a history of cerebral palsy underwent VV-ECMO for acute respiratory distress syndrome (ARDS) due to septic shock caused by bacterial translocation during treatment for HUS. He showed features of hemolysis, with elevated lactate dehydrogenase (LDH), fragmented red blood cells, and low haptoglobin levels. Plasma free hemoglobin was measured daily throughout the whole course of ECMO with levels higher than 10mg/dL but not exceeding 50mg/dL. The extracorporeal membrane oxygenation (ECMO) circuit pressures were carefully monitored to ensure the pump generated no excessive negative pressure. The patient was weaned off ECMO on the eleventh day. There have been several cases of VA-ECMO in patients with HUS; however, there is limited literature on VV-ECMO. As the days on VV-ECMO tend to be longer than those on VA-ECMO, features of hemolysis may complicate management. Although HUS did not directly influence the clinical course in the present case, features of hemolysis were continuously observed. This case highlighted the importance of standard ECMO monitoring, especially daily measurement of plasma free hemoglobin.

Aortic Valve Stenosis Causes Accumulation of Extracellular Hemoglobin and Systemic Endothelial Dysfunction.

Whether aortic valve stenosis (AS) can adversely affect systemic endothelial function independently of standard modifiable cardiovascular risk factors is unknown. We therefore investigated endothelial and cardiac function in an experimental model of AS mice devoid of standard modifiable cardiovascular risk factors and human cohorts with AS scheduled for transcatheter aortic valve replacement. Endothelial function was determined by flow-mediated dilation using ultrasound. Extracellular hemoglobin (eHb) concentrations and nitric oxide (NO) consumption were determined in blood plasma of mice and humans by ELISA and chemiluminescence. This was complemented by measurements of aortic blood flow using 4-dimensional flow acquisition by magnetic resonance imaging and computational fluid dynamics simulations. The effects of plasma and red blood cell (RBC) suspensions on vascular function were determined in transfer experiments in a murine vasorelaxation bioassay system. In mice, the induction of AS caused systemic endothelial dysfunction. In the presence of normal systolic left ventricular function and mild hypertrophy, the increase in the transvalvular gradient was associated with elevated eryptosis, increased eHb, and increased plasma NO consumption; eHb sequestration by haptoglobin restored endothelial function. Because the aortic valve orifice area in patients with AS decreased, postvalvular mechanical stress in the central ascending aorta increased. This was associated with elevated eHb, circulating RBC-derived microvesicles, eryptotic cells, lower haptoglobin levels without clinically relevant anemia, and consecutive endothelial dysfunction. Transfer experiments demonstrated that reduction of eHb by treatment with haptoglobin or elimination of fluid dynamic stress by transcatheter aortic valve replacement restored endothelial function. In patients with AS and subclinical RBC fragmentation, the remaining circulating RBCs before and after transcatheter aortic valve replacement exhibited intact membrane function, deformability, and resistance to osmotic and hypoxic stress. AS increases postvalvular swirling blood flow in the central ascending aorta, triggering RBC fragmentation with the accumulation of hemoglobin in the plasma. This increases NO consumption in blood, thereby limiting vascular NO bioavailability. Thus, AS itself promotes systemic endothelial dysfunction independent of other established risk factors. Transcatheter aortic valve replacement is capable of limiting NO scavenging and rescuing endothelial function by realigning postvalvular blood flow to near physiological patterns. URL: https://www.clinicaltrials.gov; Unique identifiers: NCT05603520 and NCT01805739.

RED CELL FRAGMENTATION SYNDROME: A REVIEW

Hemolytic anemia and perhaps fatal consequences can result from red cell fragmentation syndrome, a disorder marked by the fragmentation of red blood cells. This syndrome can occur due to various underlying causes, including abnormalities of the heart, blood vessels, or vascular malformations. This review aims to discuss the current understanding of red cell fragmentation syndrome, its causes, and its management. In this review, we will discuss the current understanding of red cell fragmentation syndrome, its causes, and its management.

#2699 Rare glomerular microangiopathy: clinicopathological characteristics and prognosis

Abstract Background and Aims Different from the typical histologic changes of thrombotic microangiopathy (TMA), we identified a group of patients whose pathology with glomerular involvement without blood vessel lesions, which was rare and known as glomerular microangiopathy (GMA). Method In Peking Union Medical College Hospital, a total of 33 patients were diagnosed as GMA with proliferation and swelling of glomerular endothelial cells, fragmented red blood cells, fibrin or platelet thrombi in the capillary lumen, thickening of the basement membrane, stenosis or occlusion of capillary lumen, but no abnormalities in the endothelium of renal small vessels. The pathology and clinical data were from the medical records and analyzed. Results The patients were 66.7% male with a mean of 45 years (14 to 74 years). The primary diseases were Castleman's disease (CD, 30.3%), POEMS syndrome (21.2%), cancer patients receiving agents targeting vascular endothelial growth factor (21.2%), and others. All the patients showed various degrees of proteinuria, with an average 24-hour urinary protein of 2.66 g/d, hematuria (63.6%), and acute kidney injury (AKI, 39.4%). The median serum creatinine (SCr) was 142 (107.5, 169) μmol/L. The AKI patients had lower Alb (P < 0.0001), PLT (P = 0.0192), higher high-sensitivity C-reactive protein (hs-CRP) (P = 0.0199), and reduction of IgA (P = 0.0021). There is no significant difference in SCr and proteinuria among patients with different etiologies. The CD patients exhibited higher hs-CRP and D-Dimer. Treatment included targeting therapy of underlying hematological disorders, glucocorticoid combination with or without immunosuppressant, and supportive treatment. After treatment, we observed proteinuria remission (66.7%), partial improvement (27.3%), or complete recovery (45.5%) of renal function. Conclusion In Castleman's disease, POEMS syndrome, and patients receiving agents targeting VEGF, initial attention to GMA might improve the prognosis and shed light on the study of the mechanisms.

Tacrolimus-Associated Thrombotic Microangiopathy Presenting with Ischemic Colitis After Kidney Transplantation: A Case Report

Calcineurin inhibitors (CNIs) are a significant component of the immunosuppressive regimen after kidney transplantation. By inhibiting cytokine gene transcription, CNIs suppress T cell and T cell-dependent B cell activation. Tacrolimus is preferred in most patients undergoing kidney transplantation. Thrombotic microangiopathy (TMA) is a severe but rare complication of CNIs therapy. TMA defines a specific pathologic lesion of arterioles and capillaries that leads to microvascular thrombosis. A 45-year-old male underwent kidney transplantation five months ago due to autosomal dominant polycystic kidney disease (ADPKD). His triple-maintenance immunosuppressive therapy includes tacrolimus. Abdominal pain and bloody diarrhea occurred in the fifth month of posttransplant. The edematous and erythematous mucosa of the ascending colon was detected on the colonoscopy. The foci of microthrombi inside the vessel lumen in the lamina propria were shown biopsy. The thrombosis or occlusion was excluded with computerized tomography (CT) angiography in abdominal vessels. The fragmented red blood cells and moderate thrombocytopenia were detected on the peripheral blood smear. Eventually, TMA diagnosis was established through laboratory and histological findings. Tacrolimus was suspected as the trigger of the pathological process and promptly switched to the everolimus. Afterward, laboratory abnormalities and clinical symptoms were improved. In this case, we intend to emphasize drug-associated TMA and atypical presentations, such as colonic microvasculature involvement.

Schistocyte detection in artificial intelligence age.

Schistocytes are fragmented red blood cells produced as a result of mechanical damage to erythrocytes, usually due to microangiopathic thrombotic diseases or mechanical factors. The early laboratory detection of schistocytes has a critical impact on the timely diagnosis, effective treatment, and positive prognosis of diseases such as thrombocytopenic purpura and hemolytic uremic syndrome. Due to the rapid development of science and technology, laboratory hematology has also advanced. The accuracy and efficiency of tests performed by fully automated hematology analyzers and fully automated morphology analyzers have been considerably improved. In recent years, substantial improvements in computing power and machine learning (ML) algorithm development have dramatically extended the limits of the potential of autonomous machines. The rapid development of machine learning and artificial intelligence (AI) has led to the iteration and upgrade of automated detection of schistocytes. However, along with significantly facilitated operation processes, AI has brought challenges. This review summarizes the progress in laboratory schistocyte detection, the relationship between schistocytes and clinical diseases, and the progress of AI in the detection of schistocytes. In addition, current challenges and possible solutions are discussed, as well as the great potential of AI techniques for schistocyte testing in peripheral blood.

Bufalin reprograms erythrocyte lifespan through p38 MAPK and Rac1 GTPase

Ample evidence indicates that bufalin (BFN), a cardiotonic steroid in Bufo toad toxin, possesses a potent anticancer activity mainly by stimulating apoptosis in cancer cells. Human red blood cells (RBCs) undergo eryptosis which contributes to a plethora of pathological conditions. No reports, however, have examined the potential toxicity of BFN to RBCs. This study aims to characterize the biochemical mechanisms governing the influence of BFN on the physiology and lifespan of RBCs.Isolated RBCs from healthy volunteers were exposed to anticancer concentrations of commercially available BFN from the skin of Bufo gargarizans (10–200 μM) for 24 h at 37 °C. Photometric assays were used to estimate hemolysis and hemolytic markers, and flow cytometry was used to detect eryptotic markers. Phosphatidylserine externalization was captured by fluorescein isothiocyante-labeled annexin V, cellular dimensions by light scatter patterns, and intracellular Ca2+ and reactive oxygen species (ROS) by fluorogenic dyes Fluo4/AM and 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), respectively. BFN caused Ca2+-independent hemolysis and release of LDH, AST, CK, and K+, and increased annexin V-bound cells, cytosolic Ca2+, cell shrinkage, and ROS levels. BFN also disrupted Na+ and Mg2+ trafficking, and was sensitive to PEG 8000, sucrose, SB203580, and NSC 23766. In whole blood, BFN depleted hemoglobin stores, increased fragmented RBCs, and was selectively toxic to reticulocytes, lymphocytes, and platelets. In conclusion, BFN elicits premature RBC death, subject to regulation by p38 MAPK and Rac1 GTPase, and is detrimental to other peripheral blood cells. Altogether, these novel findings prompt cautious consideration of the toxin in anticancer therapy.

Eculizumab Utilization in Hemolytic Uremic Syndrome (HUS) Presenting with Acute Hemolysis

Background: Hemolytic uremic syndrome (HUS) is thrombotic microangiopathy in which microthrombi are formed, consisting mainly of platelets, which will occlude the arterioles and capillaries leading to a triad of intravascular hemolysis, thrombocytopenia, and acute kidney failure. Case Presentation: A 2-year-old boy presented to our outpatient clinic with persistent diarrhea streaked with blood, vomiting, signs of dehydration, and an ingested throat. The complete blood count (CBC) revealed acute hemolysis with mild normocytic hypochromic anemia, anisopoikilocytosis, fragmented red blood cells, and moderate thrombocytopenia. The patient experienced a drastic course which was dramatically improved after the administration of eculizumab. Conclusion: Eculizumab is a recombinant humanized monoclonal antibody against complement component C5; that effectively blocks the cleavage of complement factor C5 and, subsequently, the complement membrane attacks complex C5B-9.78. While on eculizumab, our patient's kidney function improved significantly after the first dose with a creatinine of 0.56 mg/dl.

Short Prewarming at 41°C to Correct the Interference in Samples with the Presence of Cold Agglutination.

Cold agglutinins (CAs) in blood samples can cause a reversible agglutination of red blood cell (RBC) which result in an incorrect complete blood count (CBC). So, it is important to explore new simple and feasible treatment conditions for clinical work. The CAs group included 32 samples with CAs. The parameters of CBC at room temperature or after prewarming at 37°C or 41°C for different time periods were compared. The consistency and correlation of those parameters were analyzed. The morphology of erythrocytes in the CAs group was observed manually. The control group included 45 samples without CAs and prewarmed at 37°C or 41°C for different time periods. The differences were also analyzed. CAs have a significant effect on CBC. After prewarming at 37°C or 41°C the interferences are all corrected. Consider prewarming at 37°C for 120 minutes as the standard procedure. The consistency and correlation analysis showed there was no statistical difference between the results of each subgroup and standard group, except the MCHC of group 41°C 10 minutes. The correlation of parameters between all subgroups and the standard group is satisfied. Microscopic examination showed no RBC aggregation or fragmentation after prewarming at 41°C or 37°C. According to the maximum bias requirements for expert performance in Validation, Verification, and Quality Assurance of Automated Hematology Analyzers, 2nd Edition (CLSI H26-A2), the differences in overall results in control group are negligible. The 41°C 2 minutes prewarming method is a rapid and effective way for treating samples with CAs. It is an efficient way to obtain more reliable CBC results, without specific instruments.

Assessing the accuracy and clinical utility of GPT-4O in abnormal blood cell morphology recognition.

To evaluate the accuracy and clinical utility of GPT-4O in recognizing abnormal blood cell morphology, a critical component of hematologic diagnostics. GPT-4O's blood cell morphology recognition capabilities were assessed by comparing its performance with hematologists. A total of 70 images from the Chinese National Center for Clinical Laboratories, External Quality Assessment (EQA) from 2022 to 2024 were analyzed. Two experienced hematology experts evaluated GPT-4O's recognition accuracy using a Likert scale. GPT-4O achieved an overall accuracy of 70% in blood cell morphology recognition, significantly lower than the 95.42% accuracy of hematologists (p < 0.05). For peripheral blood smears and bone marrow smears, GPT-4O's accuracy was 77.14% and 62.86% respectively. Likert scale evaluations revealed further discrepancies, with GPT-4O scoring 288.50 out of 350, compared to higher manual scores. GPT-4O accurately recognized certain intracellular inclusions such as Howell-Jolly bodies and Auer rods, while it misidentified fragmented red blood cells as neutrophilic metamyelocytes and oval-shaped red blood cells as sickle cells. Additionally, GPT-4O had difficulty accurately identifying intracellular granules and distinguishing cell nuclei and cytoplasm. GPT-4O's performance in recognizing abnormal blood cell morphology is currently inadequate compared to hematologists. Despite its potential as a supplementary tool, significant improvements in its recognition algorithms and an expanded dataset are necessary for it to be reliable for clinical use. Future research should focus on enhancing GPT-4O's diagnostic accuracy and addressing its current limitations.

Multifaktorska etiologija atipičnog hemolitičko uremijskog sindroma - prikaz slučaja

Abstract: Introduction: Hemolytic uremic syndromes are characterized by the simultaneous occurrence of hemolytic anemia, microangiopathy, thrombocytopenia, and acute renal insufficiency. In terms of the clinical prodrome, they can be classified as typical, which is more common and occurs in 90% of cases, often preceded by diarrheal syndrome induced by enterohemorrhagic Escherichia coli. Alternatively, there is an atypical and rarer form associated with pneumococcal infection, dysregulation of the alternative complement pathway, and cases involving the use of cyclosporine. Hemolytic anemia is confirmed in laboratory analyses (presence of fragmented red blood cells, decreased hemoglobin, undetectable haptoglobin values, and elevated LDH values), along with thrombocytopenia and an increase in nitrogenous substances (urea and creatinine). Case report: The report details the case of an 18-month-old girl who experienced acute renal insufficiency subsequent to a respiratory infection. Ten days preceding admission, the patient exhibited nasal discharge, and during the seven days leading up to hospitalization, she presented with fever. Furthermore, two days prior to admission, the onset of persistent vomiting and abdominal pain occurred. Suspected of bowel intussusception, the patient underwent a surgical assessment where acute surgical pathology was ruled out. The absence of urination, coupled with heightened urea and creatinine levels, prompted consideration of hemolytic-uremic syndrome, later confirmed as atypical during hospitalization. This was grounded in the clinical presentation, devoid of diarrhea syndrome but marked by nasal discharge over the preceding ten days. The administration of fresh frozen plasma yielded no improvement, and there were decreased values of the C3 complement component, H factor, and reduced ADAMTS13 activity. The lack of verotoxins from enterohemorrhagic Escherichia coli further supported the diagnosis of atypical hemolytic-uremic syndrome. After the first dose of eculizumab, a terminal complement C5 component inhibitor, the girl recovered renal function and established diuresis. Conclusion: The prompt diagnosis of atypical hemolytic-uremic syndrome is challenging due to nonspecific symptoms like nasal discharge, vomiting, fatigue, and abdominal pain. Laboratory analyses, lacking specific criteria, make it difficult to conclusively identify aHUS at the disease's onset. In Serbia, pneumococcal immunization is recommended as a preventive measure, administered through a conjugated vaccine in three doses starting from the second month of life. Rapid and accurate differentiation between typical and atypical HUS is crucial for effective treatment and prognosis. Typical HUS requires hemodialysis and plasmapheresis, whereas atypical HUS is managed with plasmapheresis, immunosuppressive therapy, and eculizumab. Administering eculizumab heightens the risk of meningococcal infection by inhibiting the C5 complement component. Therefore, it is crucial not to disregard the importance of meningococcal immunization.

Comparison of Platelet Counts Estimated by Peripheral Blood Smear Examination and Automated Haematology Analyzer

Introduction: Platelet count estimation is one of the common as well as important investigations in clinical practice to diagnose many diseases like dengue, malaria, etc. Different methods for platelet estimations are available. These methods are manual counting semi-automated and automated hematology analyzer counting, platelet count estimation by peripheral blood smear (PBS) method, etc. Semi-automated or automated analyzers may produce erroneous results in the presence of particles or light scatter like giant platelets, fragmented red blood cells, and platelet clumps, so alternative methods like PBS examination can be used for validation. Aim: To compare platelet count estimation performed by the automated cell counter method and the PBS examination method. Objective: Peripheral Blood smear examination acts as a good quality control tool to validate the results produced by the automated cell counter. Materials and Methods: The present study was carried out in the Department of Pathology, Akshaya Health Centre, Urban Bangalore. The study included 100 random blood samples collected into ethylenediaminetetraacetic acid (EDTA) vacutainers over 3 months. These were analyzed by both peripheral blood smear and automated cell counter for platelet estimation. The statistical analysis was done for test performances and their comparisons by using the coefficient of variation (CV), linear regression, and mean differences with SPSS software. Results: No significant difference (p = 0.06635) was observed between the manual peripheral blood smear (PBS) method (platelets average per 100x, multiplied by 15000/ µl) of platelet estimation (207.13 ±15.898 x 1000/ µl) and that of automated cell counter platelet value (206.53 ±16.278 x 1000/ µl). A significant positive correlation was observed between the results of both methods (r=0.9995, p &lt; 0.001) when analyzed by the Pearson correlation test. Conclusion: The peripheral blood smear platelet estimation results are comparable with automated analyzer results. Hence, the PBS examination serves as a quality control tool in assessing the results of the automated cell counters.

Shiga Toxin-Producing Escherichia Coli Associated Hemolytic Uremic Syndrome

Shiga toxin-producing Escherichia coli (STEC)-associated hemolytic uremic syndrome (STEC-HUS) is a clinical syndrome involving hemolytic anemia (with fragmented red blood cells), low levels of platelets in the blood (thrombocytopenia), and acute kidney injury (AKI). It is the major infectious cause of AKI in children. Severe forms can be associated with multiorgan involvement during the acute stage of the disease. Endothelial injury is the trigger event in the microangiopathic process. The host inflammatory response to toxin and E. coli lipopolysaccharide (LPS) is involved in disease pathophysiology. Early diagnosis and identification of underlying pathogenic mechanisms are of great significance for improving prognosis and reducing sequelae and mortality. Typical management of STEC-HUS patients relies on supportive care for electrolyte and water imbalance, anemia, hypertension, and renal failure. At present there is no specific therapy to ameliorate the prognosis. The Immediate outcome is most often favorable but long-term renal sequelae are frequent due to nephron loss. This review summarizes current knowledge regarding the epidemiological findings, the pathophysiological and clinical aspects, and its diagnosis and management.

Performance evaluation of a novel platelet count parameter, hybrid platelet count, on the BC-780 automated hematology analyzer.

Automated hematology analysis is expected to improve the performance of platelet counting. We evaluated the performance of a new platelet counting, hybrid (PLT-H) and also impedance (PLT-I) and optical (PLT-O) on the BC-780 automated hematology analyzer compared to the international reference method (IRM) in blood samples with thrombocytopenic and platelet interference. The basic platelet count performance of the BC-780 automated hematology analyzer was evaluated according to the requirements of the Clinical Laboratory and Standards Institute (CLSI) Document H26-A2. Additionally, the thrombocytopenic (low PLT count) blood samples and the platelet interference blood samples including fragmented red blood cells (RBCs), microcytes or small RBCs, and giant platelets were determined with the BC-780 hematology analyzer compared to the IRM. Blank counting and the carry-over contamination rate of platelet count using the BC-780 both met the manufacturers' claim. For both 123 thrombocytopenic and 232 platelet interference blood samples (72 fragmented RBCs, 91 microcytes and 51 giant platelets), all three platelet counting methods exhibited high comparability with the IRM (the lowest correlation (r)=0.916). Interestingly, the comparability of PLT-H (r=0.928-0.986) with the IRM was better than that of PLT-I (r=0.916-0.979). The performance of PLT-H in the BC-780 met the manufacturer's specifications. PLT-H exhibits better reproducibility than did PLT-I, correlates well with the PLT-O for thrombocytopenic samples and demonstrates good anti-interference ability. PLT-H counting is therefore recommended as a zero-cost alternative platelet counting method for platelet interference samples in clinical settings.

ESTIMATION AND COMPARISON OF PLATELET COUNT: AUTOMATED ANALYZERVS MANUAL PERIPHERAL SMEAR METHOD

Accurately assessing platelet counts holds paramount importance in the realm of diagnostics and treatment. The adoption of automated methods within the diagnostic landscape is steadily growing, attributed to their multifaceted benefits. Nonetheless, automated analyzers sometimes yield erroneous results, particularly in scenarios involving particles of comparable sizes or instances of light scattering. These encompass fragmented red blood cells (RBCs), microcytic RBCs, and the presence of oversized platelets or platelet clusters. In response, the manual estimation of platelet counts using Leishmans stain validation gains significance. Method: this study encompassed the collection of blood samples from a diverse pool of 100 patients, including both inpatients and outpatients, spanning the period from 1st February 2023 to 31st July 2023. These samples, procured from GMC Doda and its affiliated hospital, were collected in ethylenediaminetetraacetic acid (EDTA) tubes. The manual platelet count was done, followed by a comprehensive comparative analysis with the counts obtained through automated platelet analysis. Results: the outcomes unveiled a nuanced trend, with manual slide-derived results slightly surpassing those generated by automated analyzers. Conclusion: This study concluded a discernible correlation between automated and manual methodologies for platelet count assessment. Nevertheless, in instances marked by exceedingly high or low platelet counts, the manual method holds its ground, ensuring precision by circumventing concerns like platelet clumping or uneven distribution.

Study on the value of fragmented red blood cell and fragmented red blood cells percentage for predicting the prognosis of sepsis patients

To explore the predictive efficacy of fragmented red blood cells (FRC) and fragmented red blood cells percentage(FRC%) with regarding for the prognosis of septic patients, along with comparing with routine coagulation parameters. A prospective study was conducted. A total of 101 patients with sepsis who met the requirements admitted to the Intensive Care Department of the Third Xiangya Hospital of Central South University from June 1, 2022 to January 10, 2023 were selected as the research objects, they were divided into survival group and death group according to the 30-day prognosis. The clinical data and laboratory indexes such as FRCs, FRC% and Platelet (PLT) were compared between the survival group and the death group. Univariate logistic regression analysis was used first, then multivariate logistic regression analysis was used to analyze the factors affecting the prognosis. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of each parameter in the regression model for the prognosis of sepsis. The results showed that the levels of FRCs, FRC%, prothrombin time (PT), activated partial thromboplastin time (APTT) and D-dimer in death group were significantly higher than those in survival group, and PLT in death group were significantly lower than those in survival group (Z or t values were -3.712,-3.793,-2.119,-2.007,-2.209,all P<0.05). FRCs or FRC% and PLT could be independent predictors of 30-day mortality. The area under the ROC curve (AUC) of PLT for predicting 30-day death in sepsis patients was 0.727 (95%CI 0.629-0.811, P<0.01), when the optimal threshold was 137 ×109/L, the sensitivity was 83.87% and the specificity was 57.14%. The AUC of FRCs for predicting 30-day death in sepsis patients was was 0.732 (95%CI 0.635-0.815, P<0.01), when the optimal threshold was 10.1×109/L, the sensitivity was 77.42%, and the specificity was 67.14%. The AUC of FRC% for predicting 30-day death in sepsis patients was 0.737 (95%CI 0.640-0.820, P<0.01), When the optimal threshold was 0.34%, the sensitivity was 77.42%, and the specificity was 65.71%. In conclusion,PLT, FRCs and FRC% have great application value in the prognosis of sepsis. When the PLT, FRCs and FRC% of sepsis patients are more than 137 ×109/L, 10.1×109/L and 0.34% respectively, it is necessary to take necessary and reasonable clinical intervention measures as soon as possible.

Isolated arterial mucoid intimal thickening lesion in early post-transplant kidney allograft biopsies.

Thrombotic microangiopathy (TMA) on kidney biopsy shows a variable combination of features: arterial mucoid intimal thickening, acellular closure of glomerular capillary loops, fragmented red blood cells, fibrin thrombi, and arterial fibrinoid necrosis. However, some early post-transplant kidney biopsies show only arterial mucoid intimal thickening. We aimed to elucidate the importance of this finding. We identified 19 biopsies showing isolated arterial mucoid intimal thickening and compared them with 22 bona fide TMA biopsies identified based on the pathological findings (excluding rejection) (2011-2020). Additionally, delayed graft function (DGF) (n=237), and no DGF (control, n=1314) groups were included for survival analysis. Seven of 19 cases with isolated arterial mucoid intimal thickening showed peripheral blood schistocytes but no other systemic features of TMA. Eight patients underwent adjustments in maintenance immunosuppression (mainly calcineurin inhibitors). None of the cases progressed to full-blown TMA on consecutive biopsies. The overall and death-censored graft survival rates in this group were comparable to the DGF group, but significantly better than the TMA group (P=.005 and .04, respectively). Isolated arterial mucoid intimal thickening in early post-transplant biopsies may be an early/mild form of TMA, probably requiring adjustment in immunosuppressive regimen. Careful exclusion of known causes of TMA, and donor-derived arterial injury are important.

Micro-Red Blood Cell, Fragmented Red Blood Cell, Platelet Distribution Width, Mean Platelet Volume, and Platelet-Large Cell Ratio on Sysmex XN Series Hematology Analyzers Can Be Used for the Reflex Test of Impedance Platelet Count in Clinical Practice.

Platelet (PLT) counting with impedance (PLT-I) is widely used but has low specificity. PLT counting with fluorescence (PLT-F), tested by the Sysmex XN series with high specificity, can be a complementary method to PLT-I. To identify red blood cell (RBC)- and PLT-related parameters as potential influencing factors for PLT-I and establish PLT reflex test rules with PLT-F. We prospectively tested both PLT-I and PLT-F in all 3480 samples. In a development data set of 3000 samples, differences between the reflex and nonreflex groups were compared and influencing factors for PLT-I were identified by logistic regression. The area under the receiver operating characteristic (ROC) curve and cutoff values were obtained by ROC curve analysis. Validation was conducted in the remaining 480 samples (validation data set). PLT-F showed comparable results with immunoplatelet counting. In logistic regression, increased micro-RBC absolute count (micro-RBC#), fragmented RBC absolute count (FRC#), PLT distribution width (PDW), mean PLT volume (MPV), PLT-large cell ratio (P-LCR), and immature PLT fraction absolute count (IPF#) were influencing factors for PLT-I. In ROC curve analysis, the cutoff values of micro-RBC#, FRC#, PDW, MPV, and P-LCR were 0.64 × 106/μL, 0.082 × 106/μL, 15.40 fL, 11.15 fL, and 33.95%, respectively. The areas under the ROC curve of micro-RBC# and FRC# were 0.77 and 0.79, respectively. Micro-RBC#, FRC#, PDW, MPV, P-LCR, and IPF# were factors affecting PLT-I. Among them, micro-RBC# and FRC# were the most impactful factors. From our study results, micro-RBC#, FRC#, MPV, PDW, and P-LCR can be used to establish reflex test rules for PLT counting in clinical work.

Clinical-Pathological Characteristics of Renal Injuries Identify Different Clusters in Patients With Antiphospholipid Antibodies

Significant heterogeneity still exists in the nomenclature of renal involvement in antiphospholipid syndrome (APS). We applied a hierarchical cluster analysis to determine subgroups of patients according to clinical, laboratory, and renal histology characteristics in a cohort of subjects with confirmed antiphospholipid antibodies (aPL) positivity and biopsy proven aPL-related renal injuries. Kidney outcomes were then assessed at 12 months. A total of 123 aPL-positive patients were included in the study (101 [82%] female, 109 [88.6%] with systemic lupus erythematosus [SLE], 14 (11.4%) with primary APS [PAPS]). Three clusters were identified. Twenty-three patients (18.7%) were included in the first cluster (cluster 1), characterized by a higher prevalence of glomerular capillary and arteriolar thrombi and fragmented red blood cells in the subendothelial space. Cluster 2 included 33 patients (26.8%) and showed a higher prevalence of fibromyointimal proliferative lesions as seen in hyperplastic vasculopathy. Cluster 3 was the largest (67 patients, mainly with SLE) and was characterized by higher prevalence of subendothelial edema, of both glomerular capillaries and arterioles. Three different clusters of patients with aPL and renal injuries emerged from our study as follows: the first, with the worst renal prognosis, was associated with features of thrombotic microangiopathy (TMA), thrombosis, triple aPL positivity and higher adjusted Global APS Score (aGAPSS) values; the second, characterized by hyperplastic vasculopathy with an intermediate prognosis, was seen more frequently in patients with cerebrovascular manifestations; and the third, more benign in terms of outcomes and with no overt association with thrombotic features, was characterized by endothelial swelling in concomitant lupus nephritis (LN).