Immune cells and inflammatory cytokines play pivotal roles in immune response against viral infections. Herein, we performed a comprehensive immunocyte profiling to delineate the variations in immunocyte populations and cytokine levels among patients infected with Omicron presenting different clinical severities. Whole blood samples were collected from a cohort of 109 participants who sought medical assistance at Peking Union Medical College Hospital between December 2022 and February 2023 in China. Study participants were categorized into three groups based on clinical severity determined by their oxygen requirements: those without the need for oxygen supplementation (No O2, n = 30), those requiring supplemental oxygen as maximal respiratory support (O2, n = 35), and those requiring assisted ventilation (VENT, n = 44). Flow cytometry was used to evaluate the levels of immune cells and cytokines. Patients requiring oxygen supplementation and those requiring assisted ventilation exhibited widespread CD28 downregulation across T-cell subsets, along with elevated expression of CD38, HLA-DR, and CD279. Further characterization revealed reduced Th1-like and Th2-like cell frequencies and increased Th17-like cell frequencies in patients requiring either oxygen supplementation or assisted ventilation. We also observed a trend toward B-cell subset alterations and changes in innate immune populations across groups. Notably, significantly higher levels of IL-6, IL-8, and IL-10, along with lower levels of IL-1β, were observed in patients with more severe disease. This study offers valuable insights into systemic immunocyte profiling among Omicron-infected patients with varying clinical severity, which may contribute to a deeper understanding of the immunopathology of COVID-19 and inform future prospective studies.

A 91-year-old man presented with a 2-year history of non-Hodgkin lymphoma with recent purpura and oedema in the lower leg for a month. His renal function was damaged with creatinine 113 μmol/L, urea 12.22 mmol/L and uric acid 549 μmol/L. His full blood count showed haemoglobin concentration 106 g/L, white blood cells 32.91 × 109/L, lymphocyte 26.15 × 109/L and platelets 136 × 109/L. Wright–Giemsa staining of peripheral blood films demonstrated multiple spherical inclusions packed in the cytoplasm and even on the nucleus of lymphoma cells (left image, ×100 objective). Furthermore, numerous spherical inclusions were observed being released from lymphoma cells and deposited as cryoglobulin between red blood cells and lymphocytes (middle and right images, ×100 objective). The presence of immunoglobulin inclusions can be found in plasma cell neoplasms and even certain B-cell lymphomas. However, the occurrence of immunoglobulin inclusion discharge leading to cryoglobulin deposition in this case is a rare phenomenon. The formation of cryoglobulin crystals may have aggravated the renal damage and led to his recent renal insufficiency. This case provides direct evidence regarding both the cellular origin of cryoglobulins and their extracellular release process. This work was funded by Beijing Natural Science Foundation (No. 7222135), and National High Level Hospital Clinical Research Funding (No. 2022-PUMCH-A-135).

ObjectiveThe purpose of this study was to evaluate the methodological comparison of reticulocytes by using the intelligent learning system Faster R-CNN, a set of reticulocyte image detection systems developed using deep neural networks.MethodsWe selected 59 EDTA-K2 anticoagulated whole blood samples and calculated the RET% using seven different Sysmex XN full-automatic hematology analyzers with Faster R-CNN in the laboratory. We compared and evaluated the methods and statistically analyzed the correlation between the various test results.ResultsThe results indicated a high degree of consistency between the seven Sysmex XN full-automatic hematology analyzers and Faster R-CNN in detecting RET%. The correlation coefficients were 0.987, 0.984, 0.986, 0.987, 0.987, 0.988, and 0.986, respectively.ConclusionWe found that the Sysmex XN full-automatic hematology analyzers in our laboratory using the Faster R-CNN system met the requirements of the methodological comparison of reticulocyte detection and this intelligent learning system can be a useful clinical tool.

Abstract Background Iron deficiency anemia (IDA) is one of the most common leading global health problems, which could cause extreme fatigue, chest pain, headache, and even neurodevelopment deterioration in children. Traditionally, assessing serum ferritin, transferrin, and iron are the tools to diagnose IDA. However, a high false-positive rate of these conventional biomarkers might be observed due to inflammation, infection, or malignancy. Reticulocyte hemoglobin equivalent (RET-He) is a new parameter that reflects the recent functional availability of iron for erythropoiesis, with the advantage of real-time results as part of automated reticulocyte analysis. The present study, therefore, aimed to investigate the utility of RET-He for the diagnosis of patients with IDA. Methods This retrospective study collected laboratory data from patients with both serum ferritin and reticulocyte results from January 2022 through December 2022, and excluded the high white blood cell count (WBC > 9380 cells/μL) and high C-reaction protein (CRP > 0.5 mg/dL). RET-He was measured by Sysmex XN-10 or XN-20 automated hematology analyzer (Sysmex, Kobe, Japan). Serum ferritin was measured by Roche Cobas e801 (Roche Diagnostics, Mannheim, Germany). Iron and total iron binding capacity (TIBC) were measured by Roche Cobas c702 (Roche Diagnostics, Mannheim, Germany). Anemia was defined as hemoglobin concentration <13 g/dL for men and < 12 g/dL for women. Iron deficiency was defined as serum ferritin <15 μg/L. Student T-tests and Pearson correlation were performed, and P-values less than 0.05 were considered statistically significant. Statistical analyses were performed with SPSS software (Version 22.0). Results A total of 2208 patient data were enrolled in this study and were classified into the IDA, iron deficiency, anemia, and control groups according to ferritin level and hemoglobin concentration. The RET-He were 20.6, 27.7, 30.4, and 32.3 pg in the IDA, iron deficiency, anemia, and control groups, respectively. The IDA group showed significantly lower RET-He levels than the control group (P < 0.001). By Pearson correlation analysis, the RET-He showed a weak positive correlation with ferritin and hemoglobin (r = 0.236, 0.274), moderate positive correlation with iron (r = 0.354), and moderate negative correlation with TIBC (r = −0.369). In addition, the average turn-around time (received-reported) of RET-He was 16.4 min, which was shorter than other biochemical markers such as ferritin, iron, and TIBC (57.7, 34.2, 34.4 min). Conclusion The RET-He could be an assessment of the iron actually used for the biosynthesis of hemoglobin and as a useful and rapid tool for diagnosing and monitoring iron deficiency anemia.

Cytomegalovirus (CMV) infection is very common in patients suffering from sepsis and may cause poor prognosis. To explore the relationship between immune status of patients with sepsis and CMV infection, we assessed T lymphocyte subtyping and other commonly used clinical parameters in patients with sepsis upon admission to the intensive care unit (ICU) and evaluated their potential impact on diagnosis and outcomes of active CMV infection. In our study, 82 of 599 patients with sepsis were diagnosed with active CMV infection. The 28-day mortality was higher in active CMV-infected than nonactive CMV-infected patients (20.7% versus 9.9%); 51of 82 active CMV-infected patients with sepsis were assessed to have CMV-DNA-negative conversion, while 31 were persistently positive for CMV DNA. Higher CD8(+)CD28(+) T-cell counts at presentation were associated with CMV-DNA-negative conversion and lower 28-day mortality. The CMV-DNA-negative conversion and 28-day mortality of active CMV-infected patients with sepsis could be predicted using cutoff values of 151 (74.5% sensitivity and 87.1% specificity) and 64.5 (52.9% sensitivity and 92.3% specificity) CD8(+)CD28(+) T cells mL(-1) at ICU admission, respectively. Higher CD8(+)CD28(+) T-cell count was significantly associated with active CMV infection, higher CMV-DNA-negative conversion and lower 28-day mortality, which may be a potential marker for early warning of active CMV infection and outcome prediction.

ObjectiveThere is a high incidence of delirium among patients with organ dysfunction undergoing cardiac surgery who need critical care. This study aimed to explore the risk factors for delirium in critically ill patients undergoing cardiac surgery and the predictive value of related risk factors.MethodsWe conducted a prospective observational study on adult critically ill patients who underwent cardiac surgery between January 2019 and August 2021. Patients were consecutively assigned to delirium and non-delirium groups. Univariate analysis and multivariate logistic analysis were used to determine the risk factors for delirium. Receiver operating characteristic curves and a nomogram were used to identify the predictive value of related risk factors.ResultsDelirium developed in 242 of 379 (63.9%) participants. Acute Physiology and Chronic Health Evaluation II (APACHE II) and Sequential Organ Failure Assessment (SOFA) scores were 14.2 ± 5.6 and 18 ± 8.4, respectively. Patients with delirium had longer cardiopulmonary bypass time (149.6 ± 59.1 vs. 126.7 ± 48.5 min, p < 0.001) and aortic cross-clamp time (98.7 ± 51.5 vs. 86.1 ± 41.6 min, p = 0.010) compared with the non-delirium group. The area under the curve was 0.824 for CD4+ T cell count and 0.862 for CD4/CD8 ratio. Multivariate analysis demonstrated that age [odds ratio (OR) 1.030, p = 0.038], duration of physical restraint (OR 1.030, p < 0.001), interleukin-6 (OR 1.001, p = 0.025), CD19+ B cell count (OR 0.996, p = 0.016), CD4+ T cell count (OR 1.005, p < 0.001) and CD4/CD8 ratio (OR 5.314, p < 0.001) were independent risk factors for delirium. A nomogram revealed that age, cardiopulmonary bypass duration, CD4+ T cell count and CD4/CD8 ratio were independent predictors of delirium.ConclusionAge, duration of physical restraint, CD4+ T cell count and CD4/CD8 ratio were reliable factors for predicting delirium in critically ill patients after cardiac surgery. The receiver operating characteristic curves and nomogram suggested a potential role for CD4+ T cells in mediating potential neuroinflammation of delirium.

Cytomegalovirus (CMV) infection is very common in patients suffering from sepsis and may cause poor prognosis. To explore the relationship between immune status of patients with sepsis and CMV infection, we assessed T lymphocyte subtyping and other commonly used clinical parameters in patients with sepsis upon admission to the intensive care unit (ICU) and evaluated their potential impact on diagnosis and outcomes of active CMV infection. In our study, 82 of 599 patients with sepsis were diagnosed with active CMV infection. The 28-day mortality was higher in active CMV–infected than nonactive CMV–infected patients (20.7% versus 9.9%); 51of 82 active CMV–infected patients with sepsis were assessed to have CMV-DNA–negative conversion, while 31 were persistently positive for CMV DNA. Higher CD8 + CD28 + T-cell counts at presentation were associated with CMV-DNA–negative conversion and lower 28-day mortality. The CMV-DNA–negative conversion and 28-day mortality of active CMV–infected patients with sepsis could be predicted using cutoff values of 151 (74.5% sensitivity and 87.1% specificity) and 64.5 (52.9% sensitivity and 92.3% specificity) CD8 + CD28 + T cells mL −1 at ICU admission, respectively. Higher CD8 + CD28 + T-cell count was significantly associated with active CMV infection, higher CMV-DNA–negative conversion and lower 28-day mortality, which may be a potential marker for early warning of active CMV infection and outcome prediction.

Nodal marginal zone lymphoma (NMZL) with peripheral blood involvement is observed in only a small proportion of patients and usually shows small-to medium-sized centrocyte-like neoplastic lymphocytes in a peripheral blood smear.1Camacho F.I. Algara P. Mollejo M. et al.Nodal marginal zone lymphoma: a heterogeneous tumor: a comprehensive analysis of a series of 27 cases.Am J Surg Pathol. 2003; 27: 762-771Crossref PubMed Scopus (86) Google Scholar, 2Inamdar K.V. Medeiros L.J. Jorgensen J.L. Amin H.M. Schlette E.J. Bone marrow involvement by marginal zone B-cell lymphomas of different types.Am J Clin Pathol. 2008; 129: 714-722Crossref PubMed Scopus (26) Google Scholar, 3Traverse-Glehen A. Felman P. Callet-Bauchu E. et al.A clinicopathological study of nodal marginal zone B-cell lymphoma. A report on 21 cases.Histopathology. 2006; 48: 162-173Crossref PubMed Scopus (82) Google Scholar This disease may present with frequent plasmacytoid differentiation or occasional monocytoid appearance; however, the granulocytoid appearance of nuclei is rarely observed. Here, we report a case of NMZL with a granulocytoid appearance of nuclei and a Pelger–Huët-like anomaly in a peripheral blood smear. A 56-year-old female presented with neck lymphadenopathy and night sweats for 4 months. Her complete blood count demonstrated haemoglobin 88 g/L, lymphocytes 22.73×109/L and platelets 156×109/L. The blood film revealed 78% abnormal lymphocytes, which mimicked granulocytic development in nuclear morphology, including metamyelocyte-like kidney bean-shaped nuclei (Fig. 1A–D), band-shaped nuclei (Fig. 1E–H) and multilobed nuclei (Fig. 1I–L). It was interesting to note that some neoplastic lymphocytes showed pseudo Pelger–Huët-like pince-nez-shaped nuclei (Fig. 1M–P). Flow cytometric analysis performed on bone marrow revealed the presence of an atypical lymphocyte population that comprised 83% of the total lymphocytes analysed. The atypical lymphocytes expressed CD19, CD22, lambda, and CD200 and partially expressed CD23 but lacked CD5, CD10, CD103, kappa, CD138, CD38, CD34, and sIgM. Molecular study revealed BIRC3 and P53 mutations. Bone marrow and lymph node biopsies confirmed the diagnosis of NMZL with bone marrow infiltration. Pelger–Huët anomaly is an inherited benign condition that is characterised by the abnormality of chromosome segregation of neutrophils leading to bilobed, peanut- or dumbbell-shaped nuclei. The classic morphological abnormality is known to be caused by mutations of the Lamin B receptor (LBR) gene, which maintains the nuclear membrane structure.4Nikolakaki E. Mylonis I. Giannakouros T. Lamin B receptor: interplay between structure, function and localization.Cells. 2017; 6: 28Crossref Google Scholar Pseudo Pelger–Huët anomaly, as a marker of granulocytic dysplasia, is only involved in neutrophils, which may occur in myelodysplastic syndrome (MDS) and acute and chronic leukaemias or can be drug-induced.5Roberts T. Linenberger M. A case of benign Pelger-Huet anomaly.Blood. 2015; 126: 693Crossref PubMed Scopus (0) Google Scholar,6Cunningham J.M. Patnaik M.M. Hammerschmidt D.E. Vercellotti G.M. Historical perspective and clinical implications of the Pelger-Huet cell.Am J Hematol. 2009; 84: 116-119Crossref PubMed Scopus (36) Google Scholar The Pelger–Huët-like anomaly of lymphocytes is an extremely uncommon phenomenon that, to the best of our knowledge, has not been reported before. In contrast to the neutrophil Pelger–Huët anomaly, which is caused by failure of nuclear segmentation, the lymphocyte Pelger–Huët-like anomaly is caused by abnormal nuclear lobulation. This work was funded by National Natural Science Foundation of China (No. 81702070 ), China; CAMS Innovation Fund for Medical Sciences (CIFMS) (No. 2018-12M-AI-002 ) and Beijing Key Clinical Specialty for Laboratory Medicine - Excellent Project (No. ZK201000 ), China. The authors state that there are no conflicts of interest to disclose.

A 69-year-old man presented with a 9-year history of repeat fingertip skin lesions with itch and pain on cold exposure. His full blood count showed haemoglobin concentration 143 g/l, white blood cells 7.7 9 10/l and platelets 221 9 10/l. Wright-Giemsa staining of peripheral blood films demonstrated numerous hexagonal crystals and more amorphous deposits (top left image, 9100 objective). The crystals were very evident after red blood cells were lysed (top right image, 940). Furthermore, cryoglobulin crystals were observed phagocytosed by neutrophils (lower images, 9100). Cryoglobulin testing confirmed the presence of a type 1 cryoglobulin (monoclonal IgG kappa) in a concentration of 7.3 g/l. Coombs’ test was positive. Bone marrow examination showed 0.5% cytologically normal plasma cells with no clonality being shown on flow cytometry. Renal and liver function tests were normal and hepatitis B and C serology was negative. Cryoglobulin decreased to 2.5 g/l and his skin lesions remitted after seven cycles of chemotherapy with bortezomib, cyclophosphamide and dexamethasone. Cryoglobulin crystallization that is clearly evident on blood film examination is uncommon. The crystals and precipitates of cryoglobulin of various sizes may be misclassified as leucocytes or platelets by automated instruments and can also induce red blood cell morphological changes. Recognition of cryoglobulin in the peripheral blood films is important to avoid misdiagnosis. Cryoglobulin crystallization accompanied by neutrophil phagocytosis is an extremely rare phenomenon, which may provide insight into a potential mechanism of clearance of cryoglobulin crystals by neutrophils. The process of cryoprecipitation is thought to be dependent on temperature and concentration, so distal extremities with low temperature and kidneys with ultrafiltration function are usually the major targets. The formation of cryoglobulin crystals may aggravate the end-organ damage.

Introduction Red blood cells (RBCs) in patients with thalassemia and iron deficiency anemia (IDA) exhibit different patterns of morphological changes. However, manual quantitative analysis of the morphological changes in the RBCs is time-consuming and subjective, limiting its use in differential diagnosis. The aim of this study was to evaluate the CellaVision Advanced RBC software as a prescreening tool for differential diagnosis of thalassemia and IDA. Methods The study cohort consisted of 54 thalassemia and 46 IDA cases in the training group and 36 thalassemia and 31 IDA patients in the validation group. The CellaVision DM96 Advanced RBC software was used to analyze the RBC morphology. Results and Conclusion Specific patterns of quantitative changes in RBC shapes were found in thalassemia and IDA patients. As a single parameter, target cell was the best morphological cell type to distinguish thalassemia from IDA, with an area under the curve (AUC) of 0.79, followed by hypochromatic cells with an AUC of 0.70. Combination of target and hypochromatic cells expressed as a ratio of the percentage of target cells to percentage of hypochromatic cells (T/H ratio) presented better differential diagnostic ability with an AUC of 0.88. A cutoff value of 1.755 for T/H ratio showed a sensitivity and specificity of 80.43% and 81.48% in the training group and 88.89% and 80.65% in the validation group, respectively. Assessment of the T/H ratio using the CellaVision Advanced RBC software represents a relatively simple and economical screening procedure for diagnostic testing of thalassemia and IDA.

A 52-year-old woman presented to our hospital with postmenopausal osteoporosis and coronary atherosclerosis. Her full blood count showed a platelet count of 73 9 10/l. A blood film of EDTA-anticoagulated blood showed platelet clumping and platelet satellitism around both neutrophils and lymphocytes. More importantly, platelet phagocytosis was noted not only in neutrophils and monocytes, but also in lymphocytes (image). Platelet satellitism was absent in both citrated and heparinised blood specimens. However, platelet clumping was more obvious in heparinised blood and platelet phagocytosis by neutrophils was occasionally seen in citrated blood. A finger-prick blood film showed no platelet satellitism or phagocytosis. The corrected platelet count was 158 9 10/l. Flow cytometry for platelet-associated immunoglobulin was strongly positive with platelets from EDTA-anticoagulated blood but not with those from citrated blood. Platelet phagocytosis is an in vitro phenomenon usually involving neutrophils and occasionally monocytes. It is quite uncommon for lymphocytes, which have usually been thought to lack phagocytic ability. We postulate that the phenomenon of platelet satellitism and phagocytosis in this patient relates to immunoglobulin bound to platelets in the presence of EDTA.

![Figure][1] A 70-year-old woman presented with fatigue and splenomegaly. She had an 8-year history of polycythemia vera and had been diagnosed with secondary hemolytic anemia 2 years previously. Molecular analysis demonstrated a JAK2 V617F mutation. Laboratory testing revealed the following

Hyperhomocysteinemia (HHcy) can result from liver disease or dysfunction and further alters intracellular lipid metabolism. Cytochrome P450 (CYP) arachidonic acid epoxygenases are expressed in human cancers and promote cancer metastasis. This study explored the interaction of Hcy and CYP450 metabolism in hepatocellular carcinoma (HCC). The levels of 4-epoxyeicosatrienoic acid (EET) isomers and their generative enzyme CYP2J2 level as well as intracellular Hcy level were higher in 42 cases of HCC than in paired non-tumor tissue. Elevated Hcy-decreased DNA methylation on SP1/AP1 binding motifs and enhancement on the CYP2J2 promoter via ERK1/2 signaling was essential for CYP2J2 upregulation and EET metabolism. Increased Hcy level enhanced the neoplastic cellular phenotype, which was reversed by CYP2J2 knockdown in vitro. Furthermore, tumor growth and size as well as patterns of CYP2J2 expression and DNA demethylation were increased with HHcy in mice induced orthotopically by 2% (wt/wt) L-methionine with or without folate deficiency. Moreover, the effect was attenuated by shRNA knockdown of CYP2J2. Thus, HHcy results from but can also promote hepatocarcingenesis via CYP450-EET metabolism by crosstalk of DNA demethylation of CYP2J2 and ERK1/2 signaling.

The understanding of genetic basis for Philadelphia-negative myeloproliferative neoplasm (MPN) has got much progress in recent years. But the effect of CALR vs. JAK2V617F mutations on the clinical progression and prognosis of primary fibrosis (PMF) remains relatively obscure. In this meta-analysis, we searched Pubmed, Embase, and Web of Science databases for observational studies published until February 2016. Researches that evaluated CALR vs. JAK2V617F mutations on PMF-relevant complications (splenomegaly, leukemic transformation, or thrombosis) and overall survival were selected. Pooled adjust odds ratio (OR), hazard risk (HR), and the corresponding 95 % confidence intervals (CI) were calculated for the CALR-mutant versus the JAK2-mutant categories. Twelve studies involving 435 CALR-mutated and 1116 JAK2V617F PMF patients were analyzed. CALR-mutated patients displayed a lower risk of splenomegaly (OR 0.47, 95 % CI 0.29–0.78) and thrombosis (OR 0.52, 95 % CI 0.29–0.92) but showed no significant difference in the risk of leukemic transformation (OR 0.90, 95 % CI 0.55–1.47) when compared with JAK2-mutated patients. CALR mutation favorably affected overall survival while JAK2 mutation led to poorer survival rate (HR 2.58, 95 % CI 2.08–3.20). This meta-analysis confirmed that a genetic classification of PMF by CALR and JAK2 mutations carried significant prognostic relevance.

To improve the identification for CTCs with weak or negative CK and diploid CTCs in pancreatic cancer, we combined immune‐staining of CK, CD45, DAPI and fluorescence in situ hybridization with the centromere of chromosome 8 (CEP8) probe method. CTCs in 3.75 mL of blood were depleted for CD45 positive cells with anti‐CD45 antibodies and identified by combining CK, CD45, DAPI and CEP8 in 61 cases including 22 pancreatic cancers, 3 borderline pancreatic solid pseudopapillary tumors, 6 pancreatic benign tumors, and 30 healthy individuals. We found that enriched cells could be classified into 5 patterns: CK+CD45‐DAPI+CEP8=2 (2 hybridization signals), CK+CD45‐DAPI+CEP8>2 (>2 hybridization signals), CK‐CD45‐DAPI+CEP8>2, CK‐CD45‐DAPI+CEP8=2, and CK+/‐CD45+DAPI+CEP8=2 or >2. Among 22 pancreatic cancers, CK+CD45‐DAPI+CEP8=2 and CK+CD45‐DAPI+CEP8>2 patterns were identified in two cases, and CK‐CD45‐ DAPI+CEP8>2 pattern was identified in 16 cases. CK‐CD45‐DAPI+CEP8=2 and CK+/‐CD45+DAPI+CEP8=2 or >2 patterns were detected in pancreatic cancers, other pancreatic diseases and healthy individuals. Among the five patterns, CK+CD45‐DAPI+CEP8=2, CK+CD45‐DAPI+CEP8>2 and CK‐CD45‐DAPI+CEP8>2 were considered as CTCs, while CK‐CD45‐DAPI+CEP8=2 and CK+/‐CD45+DAPI+CEP8=2 or >2 were considered as indeterminate cells. When the cutoff value was set as 2 cells/3.75 mL based on ROC curve, the sensitivity and specificity in the diagnosis of pancreatic cancer was 68.18 and 94.87%, respectively. Dynamically monitoring CTCs changes prior to and after surgery in pancreatic patients revealed that CTCs count decreased in 3 days after surgery, but increased in 10 days after surgery in most patients. During our one and a half year follow‐up, CTCs positive patients showed metastasis and worse survival rate.

Background: This study intends to develop the microscopic review criteria for automated urine chemistry analyzer and integrated urine chemistry and formed-element analyzer.Methods: A total of 1058 samples were analyzed using chemistry analyzer (Siemens Atlas) for proteins (PRO), blood (BLD) and WBC. Cast, RBC and WBC were analyzed using 4 different instruments, IRIS IQ200, AVE-766, US 2026, and Sysmex UF-1000i. A phase-contrast microscopy was used as reference to evaluate false-negative rate (FNR) and review rate (RR).Results: The optimized review criteria for Atlas were either PRO 2+, or BLD 2+, or WBC 2+, or specimen from nephrology department. FNR was 4.65% and RR was 40.41%. The optimized criteria for integrated Atlas and IQ200, or AVE-766 or US 2026 or UF-1000i were either BLD >= 2 + (>2 + for females), or RBC count >= 2 times reference range, or different WBC results between chemistry and formed-element analysis, or PRO >= 2 + or CAST > the reference range. One additional rule for integrated Atlas and UP-1000i was different results between BLD and RBC counts. FNR was 1.94%, 2.03%, 1.74%, 1.65%, and RR was 41.09%, 40.12%, 44.86%, 50.39%, respectively.Conclusions: These specific review criteria ensured a low missed-diagnosis rate and a reasonable workload in practice. (C) 2015 Elsevier B.V. All rights reserved.

Background: CD45 is universally expressed on nucleated hematopoietic cells in healthy individuals with variable intensities. Although down-regulation of CD45 expression is commonly seen in immature B-cell neoplasm, loss of CD45 expression is rarely observed in mature B-cell lymphoproliferative disorders.Case report: A 64-y old man was presented with fatigue and night sweat in July 2013. Physical examination revealed right axillary lymphadenopathy without splenohepatomegaly. The lymph node was sent for flow cytometry and histomorphologic evaluation. Flow cytometric analysis revealed the presence of CD45 + and CD45 - cell populations. The CD45 + population (42%) comprised mainly unremarkable CD3 + T cells. However, the CD45 - population (35%) showed CD19 +, CD20 +, and CD10 + immunophenotype with monotypic surface Lambda immunoglobulin light chain expression. Immunohistochemical studies performed on tissue section revealed that neoplastic lymphoid nodules were positive for CD20, BCL-6 and BCL-2 with low proliferation index by Ki-67 staining, but again negative for CD45. FISH analysis confirmed the presence of IGH/BCL2 rearrangement Follicular lymphoma (grade 1) with negative CD45 expression was diagnosed.Conclusions: Loss of CD45 expression has been reported, but all cases documented were large B cell lymphoma (LBCL). To the best of our knowledge, this is the first case of low grade follicular lymphoma with lack of CD45 expression. We report this case because of not only its rarity but also to raise awareness that loss of CD45 expression can occur in LBCL as well as in low grade B-cell lymphomas. (C) 2015 Elsevier B.V. All rights reserved.

BACKGROUND:Conventional cytokeratin (CK)-based methods to detect circulating tumor cells (CTCs) often present suboptimal sensitivities for decreased or non-expression of cytokeratin in some CTCs. We clinically investigated a new method that combines immunocytochemistry staining (ICC) of CD45 and fluorescence in situ hybridization (FISH). METHODS:Circulating epithelial cells from 141 subjects were enriched using an EpCAM-independent strategy and then identified by either a combination of FISH with chromosome 8 centromere probe (CEP8) and ICC staining of CD45 (CD45-FISH) or ICC staining of CK. RESULTS:For detecting CTCs enriched from lung cancers, CD45-FISH had larger areas under ROC curves of 0.963 (P=0.000) compared to ICC (0.653; P=0.031) using cut-off values of 2 and 1 cell/3.75ml blood with sensitivities of 83.3% and 43.3%, specificities of 98.6% and 89.5%, respectively. Moreover, CD45-FISH showed 76.2% sensitivity in detecting CTCs in ovarian cancers (P<0.001). Four of six ovarian cancers showed dramatical decrease in both CTCs and serum CA125 on the 7th day after surgery. CONCLUSION:CD45-FISH method had improved sensitivity and specificity in detecting CTCs of lung and ovarian cancers compared to ICC-CK. This combined detection strategy may be useful in detecting or monitoring CTCs after ovarian cancer surgery.