Role of Flow Cytometry in the Diagnosis of Chronic Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To analyze the immunological phenotype of chronic myeloid leukemia (CML), and explore its characteristics and significance.@*METHODS@#The immunophenotypes of 40 CML children and 40 controls were analyzed by multicolor flow cytometry. CD45/SSC, as the basic gate, was used to delineate neutrophils. Then, the distribution of cluster differentiation (CD) molecules on the surface of granulocytes was analyzed in three ranges (≥1%, ≥5%, and ≥20%), and the expression rates of CD molecules (≥1% included in the statistical analysis) and the mean fluorescence intensity (MFI) were compared between the two groups.@*RESULTS@#The proportion of granulocytes in the CML group was (82.1±6.4)%, which was significantly higher than (57.8±11.8)% in the control group (P <0.001). The expression rates of CD15/CD11b/CD33/CD13 in CML and control groups were high, and both distributed in the range of ≥20%. The differentiation trajectory of CD33/CD13 was normal and there were no significant differences in the expression rate and MFI between the two groups. However, both the expression rate of CD11b and CD15 MFI in the CML group were significantly lower than those in the control group (P <0.001). There were no significant differences in the expression rate and MFI of CD10 between the two groups, and the expression levels of CD10 between the two groups were consistent in different distributions. In the CML group, there was a large number of cases with abnormal high expression of CD56, 52.5% of the cases had a CD56 expression rate of ≥5%, and 42.5% had a CD56 expression rate of ≥20%, while the control group did not express CD56 (<1%). The expression distribution of CD117 was different between the two groups. In the range of expression rate ≥5%, there were 35.0% cases in the CML group, while only 2.5% in the control group. The expression rate of CD117 in the CML group was higher than that in the control group (P <0.001), though there was no significant difference in MFI.@*CONCLUSION@#The immunophenotyping of CML is characterized by increased proportion of mature neutrophils, decreased CD15 MFI, decreased proportion of CD11b and abnormal high expression of CD56 and CD117. Flow cytometric analysis of immunophenotype can effectively distinguish normal granulocytes from chronic granulocytes, and help in the diagnosis of CML.

Possible Risk Factors for Bone Marrow Fibroplasia in Patients with Polycythemia Vera / 中国实验血液学杂志

OBJECTIVE@#To understand the biological characteristics of polycythemia vera (PV) patients with myeloid fibroplasia, and further analyze the risk factors affecting myeloid fibroplasia in PV patients, so as to provide ideas for predicting the occurrence of myeloid fibroplasia in PV patients.@*METHODS@#Forty patients with PV in the Department of Hematology, Xiyuan Hospital of China Academy of Chinese Medical Sciences were collected and divided into two groups, with (hyperplasia group) and without (Non-proliferative group) hyperplasia of bone marrow fibers. The differences of basic clinical characteristics, blood routine, biochemistry, bone marrow cells, coagulation function and other indicators between the two groups were compared, and the independent risk factors affecting the proliferation of bone marrow fibrous tissue in PV patients were further analyzed by multivariate regression.@*RESULTS@#Compared with Non-proliferative group, the JAK2 mutation rate (95% vs 70%，P=0.037), eosinophilic cell count (0.19 vs 0.11, P=0.047) and eosinophilic percentage (1.84 vs 1.27, P=0.001) in PV patients with hyperplasia were significantly increased, triglycerides (1.55 vs 1.91, P=0.038) and low-density lipoprotein (1.50 vs 3.08, P=0.000) were significantly reduced, bone marrow hematopoietic volume (0.85 vs 0.6, P=0.001), granulocyte/erythrocyte ratio (3.40 vs 1.89, P=0.033), lymphocyte/erythrocyte ratio (0.60 vs 0.42, P=0.033), and granulocyte+lymphocyte/erythrocyte ratio (3.72 vs 2.37, P=0.026) were significantly increased, thrombin time (18.84 vs 18.12, P=0.043) was significantly prolonged. Multivariate regression analysis results showed that peripheral blood eosinophil ≥2% and low-density lipoprotein ≤2 mmol/L were independent risk factors for bone marrow fibrous tissue hyperplasia in PV patients (P<0.05).@*CONCLUSION@#Increased proportion of peripheral blood eosinophils and decreased low density lipoprotein are risk factors for bone marrow fibrous tissue hyperplasia in PV patients.

Echocardiographic epicardial fat thickness and immature granulocyte are novel inflammatory predictors of acute ischemic stroke: a prospective study

ABSTRACT BACKGROUND: Acute ischemic stroke (AIS) is the most common type of stroke. Inflammation is the primary factor in the pathogenesis of atherosclerosis. Use of immature granulocytes (IGs) has been recommended as a new indicator of systemic inflammation. However, data on the association between echocardiographic epicardial fat tissue thickness (EFT) and IGs in patients with AIS are limited. OBJECTIVE: To evaluate the association between the presences of IGs, epicardial fat tissue and AIS. DESIGN AND SETTING: Prospective study in a tertiary-care university hospital in Antalya, Turkey. METHODS: Our study included 53 AIS patients and 41 healthy controls with age and gender compatibility. Blood samples and transthoracic echocardiography of all participants were compared. RESULTS: IG levels were significantly higher in patients with AIS than in controls (0.62 ± 0.36 versus 0.28 ± 0.02, P < 0.001). The mean EFT was 3.74 ± 0.61 mm in the control group and 6.33 ± 1.47 mm in the AIS patient group. EFT was significantly greater in AIS patients than in controls (P < 0.001). For the optimum cut-off value for IG (0.95), the area under the curve (AUC) was determined to be 0.840; sensitivity was determined to be 81.1% and specificity, 92.5%. For the optimum cut-off value for EFT (4.95 mm), the AUC was determined to be 0.953; sensitivity was determined to be 90.6% and specificity, 90%. CONCLUSIONS: IG and echocardiographic EFT are clinical markers that can be used to predict AIS risk.

Nelore females along a new Brazilian agricultural frontier: hematological and clinical-biochemical approaches / [Abordagens hematológicas e bioquímicas clínicas de fêmeas Nelore na nova fronteira agrícola brasileira]

The Nelore breed is the second largest bovine breed in the world and has actively participated in the expansion of new Brazilian agricultural frontiers. In this context, the purpose of this study was to determine the hematological and biochemical reference intervals of healthy Nelore matrices raised under an extensive regime without supplementation along southwest of Piauí state. Blood samples were collected from fifty-five multiparous female of the Nelore breed. Biochemical and hematological parameters were analyzed using a parametric statistical method with 95% CI of reference limits. The average values of red blood cells, hemoglobin as well as hematimetric indices showed reference ranges similar to reference standards. The hematocrit as well as granulocytes and agranulocytes presented alterations typical of animals raised in environments with higher temperatures. Mineral, enzymatic, protein and metabolic profiles were similar to other bovine breeds but with a narrower range of values. However, lower mean values were observed for levels of ionized calcium, total protein and urea. Nelore females present slightly different biochemical and hematological profiles from other breeds, which might result from the environmental and nutritional management applied and the natural deficiency of nitrogen, phosphorus and calcium in the region's pastures.(AU)

Nelore é a segunda maior raça bovina do mundo e tem participado ativamente da expansão das novas fronteiras agrícolas brasileiras. Nesse contexto, o objetivo do presente estudo foi determinar os intervalos de referência hematológicos e bioquímicos de matrizes Nelore criadas em regime extensivo sem suplementação, ao longo do sudoeste do estado do Piauí. Amostras de sangue foram coletadas de 55 fêmeas multíparas da raça Nelore. Os parâmetros bioquímicos e hematológicos foram analisados por método estatístico paramétrico com IC 95% para os limites de referência. Os valores médios de hemácias, hemoglobina e índices hematimétricos apresentaram intervalos de referência semelhantes aos padrões de referência. Tanto o hematócrito quanto os granulócitos e os agranulócitos apresentaram alterações típicas de animais criados em ambientes com temperaturas mais elevadas. Os perfis mineral, enzimático, proteico e metabólico foram semelhantes aos de outras raças bovinas, mas com uma faixa de valores mais estreita. No entanto, valores médios mais baixos foram observados para os níveis de cálcio ionizado, proteína total e ureia. Fêmeas Nelore apresentam perfis bioquímicos e hematológicos ligeiramente diferentes de outras raças, o que pode resultar dos manejos ambiental e nutricional aplicados e da deficiência natural de nitrogênio, fósforo e cálcio nas pastagens da região.(AU)

Effect of PADI4 on the Expression of Inflammatory Cytokines During NB4 Cells Differentiation / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of peptidylarginine deiminase 4 (PADI4) during the process of differentiation into granulocyte of NB4 cells induced by all-trans-retinoic acid (ATRA) and whether PADI4 is involved in the inflammatory cytokines expression.@*METHODS@#Granulocyte differentiation model of NB4 cells induced by ATRA was established. The cell morphology changes were observed by Wright-Giemsa staining. The expression of cell differentiation marker CD11b was analyzed by flow cytometry. The mRNA and protein expression of PADI4 was detected by RT-PCR and Western blot, respectively. The expression of tumor necrosis factor (TNF) α and interleukin (IL) 1β was analyzed by ELISA, and also examined with the knockdown of PADI4 expression by siRNA.@*RESULTS@#After NB4 cells induced by ATRA, the cytoplasm increased and the ratio of nuclear to cytoplasmic was reduced. Nuclear dented, and rod-shaped nucleus, lobulated phenomenon increased (P<0.05). Flow cytometry analysis results showed that the cell surface molecule CD11b expression increased (P<0.01). RT-PCR and Western blot showed the expression of PADI4 increased at both transcriptional and translational levels during the process of the differentiation. ELISA showed TNF-α and IL-1β secretion increased in differentiated macrophages, while they could be inhibited by PADI4-specific siRNA.@*CONCLUSION@#During the differentiation into granulocyte of NB4 cells induced by ATRA, PADI4 expression increased. Furthermore, PADI4 appeared to play a critical role in inflammatory cytokines secretion.

A Case of Cutaneous Leukocytoclastic Vasculitis Associated with Granulocyte Colony-Stimulating Factor: An Unusual Presentation

Drug-induced vasculitis is an inflammation of small-sized blood vessel caused by the use of drugs. It accounts for approximately 10% of acute cutaneous vasculitis. Propylthiouracil, hydralazine, and allopurinol have been widely known as causative agents. The most common clinical feature of drug-induced vasculitis is palpable purpura on lower extremities. A 66-year-old Korean female presented with erythematous nodules on upper chest and back. She had been on medication for multiple myeloma. Laboratory results showed neutropenia. After a single injection of filgrastim (recombinant granulocyte colony-stimulating factor), she developed cutaneous lesions with concurrent increase in absolute neutrophil count. A skin biopsy revealed leukocytoclastic vasculitis. After discontinuation of filgrastim injection, her skin lesions disappeared spontaneously.

Myelodysplastic syndrome: validation of flow cytometry multilineage score system / Síndrome mielodisplásica: validação de ficha de escore multilinhagem por citometria de fluxo

ABSTRACT Objective To validate multilineage score system correlating results of flow cytometry, cytogenetics, cytomorphology and histology from samples of patients with suspected myelodysplastic syndrome or cytopenia of unknown origin. Methods A retrospective study analyzing laboratory data of 49 patients with suspected myelodysplastic syndrome or cytopenia of unknown origin, carried out between May and September 2017. The inclusion criteria were availability of flow cytometry results, and at least one more method, such as morphology, histology or cytogenetics. Thirty-eight patients were classified as diagnosis of myelodysplastic syndromes, whereas 11 were classified as normal. Patients were evaluated based on score systems, Ogata score and flow cytometry multilineage score. Results Comparing the scores obtained in the Ogata score and the multilineage score, it was observed that in four cases the Ogata score was zero or 1 point, while the multilineage score was higher than 3 points. In addition, in 12 cases with Ogata score of 2, the multilineage score was greater than 3. Conclusion The flow cytometry multilineage score system demonstrated to be more effective in dysplasia analysis, by assessing the erythroid, monocytic, granulocytic and precursor cell lineages, apart from the parameters evaluated by the Ogata score.

RESUMO Objetivo Validar ficha de escore multilinhagem correlacionando resultados obtidos de citometria de fluxo, citogenética, citomorfologia e histologia de amostras de pacientes com suspeita de síndrome mielodisplásica ou citopenias a esclarecer. Métodos Estudo retrospectivo de análise de dados laboratoriais de 49 pacientes com suspeita clínica de síndrome mielodisplásica ou citopenias a esclarecer realizado entre maio e setembro de 2017. Os critérios de inclusão foram a disponibilidade de resultados de citometria de fluxo e de, pelo menos, outra metodologia, entre morfologia, histologia, ou citogenética. Trinta e oito pacientes foram classificados como diagnosticados com síndromes mielodisplásicas enquanto 11 foram classificados como normais. Os pacientes foram avaliados utilizando sistemas de escore, escore de Ogata e ficha multilinhagem. Resultados Comparando as pontuações obtidas no escore de Ogata e na ficha multilinhagem, observou-se que, em quatro casos, o score de Ogata foi zero ou 1 ponto, enquanto, pela ficha multilinhagem, a pontuação foi superior a 3 pontos. Além disso, em 12 casos com escore de Ogata 2, a pontuação pela ficha multilinhagem foi superior a 3. Conclusão A ficha multilinhagem demonstrou ser mais eficaz na análise de displasia por avaliar as linhagens eritroide, monocítica, granulocítica e células precursoras, além dos parâmetros avaliados no escore de Ogata.

Analysis of hematologic parameters of donors, patients, and granulocyte concentrates to predict successful granulocyte transfusion

BACKGROUND: Granulocyte transfusion (GTx) is performed as a supportive therapy in severe neutropenic patients caused by various conditions. The study aimed to analyze the hematologic parameters of donors, patients, and granulocyte concentrates to predict successful GTx. METHODS: This study was performed in 281 donors, with their granulocyte concentrates being collected through apheresis, and in 54 severe neutropenic patients who had various hematologic diseases. Complete blood cell counts of donors pre- and post-apheresis, granulocyte concentrates, and patients pre- and post-GTx were analyzed. Patients were divided into two groups according to survival at discharge (Group S, survival; Group D, dead) to compare various factors including age, infection status, pre- and post-GTx total white blood cell counts (TWBCC) and absolute neutrophil counts (ANC), total number of GTx, infused TWBCC and ANC per weight, and use of G-CSF during therapy. RESULTS: Overall data of patients showed that both TWBCC and ANC were significantly increased after GTx (median values at pre-GTx, TWBCC=0.40×109/L, ANC=0.14×109/L; post-GTx, TWBCC=0.57×109/L, ANC=0.29×109/L, both P<0.0001). After GTx, Group S (N=25) showed significantly higher TWBCC and ANC than Group D (N=29) (P=0.01 and P=0.04, respectively). Using different cutoff levels, post-GTx TWBCC greater than 0.5×109/L showed statistically significant difference between the two groups (P<0.01). None of the other factors showed statistically significant differences. CONCLUSION: The TWBCC and ANC after GTx were significant factors to predict patients' outcome. Therefore, follow-up of those two parameters may be helpful to select or consider other therapeutic modalities including additional GTx.

Evaluation of Time and Temperature Stability of Paroxysmal Nocturnal Hemoglobinuria Cells by Flow Cytometry

BACKGROUND: Flow cytometry analysis of paroxysmal nocturnal hemoglobinuria (PNH) is significantly affected by the methodology used. The lack of data on the effect of age and refrigeration on PNH clone stability motivated us to study these aspects using flow cytometry. METHODS: Peripheral blood was collected from six patients, of which two presented with PNH. All samples were tested immediately and stored at room temperature (RT, 20–25℃) and at 4℃ for re-analysis at 24, 48, 72 hr and 7 days. Anti-CD59-fluorescein isothiocyanate (Beckman Coulter, USA) and anti-CD235a-phycoerythrin (PE; Beckman Coulter) were used to stain red blood cells (RBCs). Fluorescein-labeled proaerolysin (Cedarlane, Canada), anti-CD15-PE (Beckman Coulter), anti-CD24-PE-cyanin 5 (Beckman Coulter), and anti-CD45-PE-cyanin 7 (Beckman Coulter) were used to stain granulocytes. Flow cytometry was performed using a FC500 flow cytometer (Beckman Coulter). The effects of time and temperature were analyzed using generalized estimating equations. RESULTS: No significant differences in the gated percentage of RBCs and PNH clone size of RBCs were observed between the RT and 4℃ groups up to 7 days of testing. The percentage of gated neutrophils decreased with specimen age (P<0.001) and a better correlation with baseline was obtained at 4℃ than at RT (P=0.014). Neutrophil PNH clones were stable until 48 hr and 72 hr at RT and 4℃, respectively, and could not be analyzed at 7 days. CONCLUSIONS: RBC analysis was successfully performed up to 7 days. For neutrophils, testing within 48 hr is recommended, because the number of gated cells decreases significantly with age.

Comparison of the virulence of Streptococcus pneumoniae in ICR mouse stocks of three different origins / 한국실험동물학회지

Streptococcus pneumoniae causes many people to suffer from pneumonia, septicemia, and other diseases worldwide. To identify the difference in susceptibility of and treatment efficacy against S. pneumoniae in three ICR mouse stocks (Korl: ICR, A:ICR, and B:ICR) with different origins, mice were infected with 2 × 106, 2×107, and 2×108 CFU of S. pneumoniae D39 intratracheally. The survival of mice was observed until three weeks after the infection. The three stocks of mice showed no significant survival rate difference at 2 × 106 and 2 × 107 CFU. However, the lung and spleen weight in the A:ICR stock was significantly different from that in the other two stocks, whereas the liver weight in B:ICR stock was significantly lower than that in the other two stocks. Interestingly, no significant CFU difference in the organs was observed between the ICR stocks. The level of interferon gamma inducible protein 10 in Korl:ICR was significantly lower than that in the other two stocks. The level of granulocyte colony stimulating factor in B:ICR was significantly lower than in the other two stocks. However, tumor-necrosis factor-alpha and interleukin-6 levels showed no significant difference between the ICR stocks. In the vancomycin efficacy test after the S. pneumoniae infection, both the single-dose and double-dose vancomycin-treated groups showed a significantly better survival rate than the control group. There was no significant survival difference between the three stocks. These data showed that Korl:ICR, A:ICR, and B:ICR have no susceptibility difference to the S. pneumoniae D39 serotype 2.

Delta Neutrophil Index as an Early Marker of Sepsis in Burn Patients

Hematopoiesis Activity of Sambucus javanica on Chloramphenicol-induced Aplastic Anemia Mouse Model

Hematopoiesis has a pivotal role in the maintenance of body homeostasis. Ironically, several hematological disorder caused by chemicals, drugs, and other environmental factors lead to severe bone marrow failure. Current treatments like stem cell transplantation and immunosuppression remain ineffective to ameliorate this diseases. Therefore, a newtreatment to overcome this entity is necessary, one of them by promoting the usage of medicinal plants. Thus, this study aimed to evaluate the hematopoiesis potency of S. javanica berries and leaves extracts in chloramphenicol (CMP)-induced aplastic anemia mice model. In this present study, several types of blood progenitor cell such as TER-119⁺VLA-4⁺ erythrocytes lineage, Gr-1⁺ granulocytes, and B220⁺ B-cell progenitor cells were evaluated by flow cytometry analysis. Accordingly, we revealed that S. javanica berries and leaves extracts significantly promoted TER-119⁺VLA-4⁺ erythrocytes lineage and Gr-1⁺ granulocytes after exposed by CMP. Thus, these results suggested that S. javanica berries and leaves extracts might have hematopoiesis activity in CMP-induced aplastic anemia mice model.

Bacteriological, cytological and histopathological evaluation of the reproductive tract of slaughtered cows / Avaliação bacteriológica, citológica e histopatológica do trato reprodutivo de vacas abatidas

Reproductive diseases, mainly endometritis, are important hurdles in cattle raising, In the current study we evaluated gross, bacteriological, cytological, and histological findings from selected sites of the genital from 23 slaughtered cows and tested whether there is an association between these findings and the probability of reaching a reliable diagnosis. The results from the examinations of macroscopic aspects of uterine secretions, the cytological, bacteriological, and histopathological findings were then correlated. There was no significant correlation (P>0.05) of the statistical data from different parts of the genital tract. Trueperella pyogenes and Escherichia coli were isolated from the vagina in 3/23 cases. In only 2/23 samples Enterococcus faecalis and a gram-negative, oxidase-positive bacteria were isolated from the cervix uteri. Only Staphylococcus epidermidis, in 1/23 case, was isolated from the uterus. Histopathological findings in uterus from samples of Groups II (moderate lesions) and III (severe lesions) did not translated in grossly visible changes. Samples from reproductive tracts with secretion in the vagina and cervix uteri had no detectable changes in the other parameters analyzed from this portion. Uterus with positive bacterial culture had evidence of ascendant inflammation judging by the high granulocyte count in the three analyzed portions. This study reinforces that vaginitis and cervicitis in the cow diagnosed only by clinical examination does not reflect the real status of the uterine health. For this reason, treatment of uterine disorders should be conducted based on reliable tests to determine the appropriate therapy for each situation.(AU)

Doenças reprodutivas causam altas perdas econômicas nos rebanhos bovinos, sendo a endometrite uma das alterações mais relevantes. Os objetivos desta pesquisa foram avaliar a associação dos achados fisiopatológicos em exames macroscópicos, bacteriológicos, citológicos e histopatológicos nas porções do trato genital de 23 vacas abatidas, bem como avaliar a necessidade de associação destes exames para efetuar diagnóstico fidedigno. A avaliação macroscópica da secreção, os exames histopatológico, citológico e as bactérias identificadas foram correlacionados. Não houve associação (P>0,05) dos resultados nas diferentes porções do trato genital. Na vagina foram isoladas as bactérias Trueperella pyogenes e Escherichia coli. Na cérvix, em apenas 2/23 (8,6%) amostras isolou-se Enterococcus faecalis e gram negativo oxidase positiva. No útero houve isolamento apenas da bactéria Staphylococcus epidermidis. As amostras histopatológicas classificadas em grupo II e III não apresentaram alterações detectadas no exame macroscópico. As amostras com secreção não fisiológica na vagina e cérvix não apresentaram alterações nos outros exames na porção uterina. A amostra com cultura bacteriológica positiva no útero demonstrou uma infecção ascendente através da alta contagem de granulócitos nas três porções analisadas. O presente estudo reforça o conceito de que a vaginite e cervicite diagnosticadas pelo exame clinico na vaca não retratam o real status da saúde uterina e por esta razão, o tratamento do útero deve ser conduzido com critério, alicerçado nos exames complementares para definir a terapia adequada para cada situação.(AU)

Antiviral and Anti-Inflammatory Activities of Pochonin D, a Heat Shock Protein 90 Inhibitor, against Rhinovirus Infection

Human rhinoviruses (HRV) are one of the major causes of common cold in humans and are also associated with acute asthma and bronchial illness. Heat-shock protein 90 (Hsp90), a molecular chaperone, is an important host factor for the replication of single-strand RNA viruses. In the current study, we examined the effect of the Hsp90 inhibitor pochonin D, in vitro and in vivo, using a murine model of human rhinovirus type 1B (HRV1B) infection. Our data suggested that Hsp90 inhibition significantly reduced the inflammatory cytokine production and lung damage caused by HRV1B infection. The viral titer was significantly lowered in HRV1B-infected lungs and in Hela cells upon treatment with pochonin D. Infiltration of innate immune cells including granulocytes and monocytes was also reduced in the bronchoalveolar lavage (BAL) by pochonin D treatment after HRV1B infection. Histological analysis of the lung and respiratory tract showed that pochonin D protected the mice from HRV1B infection. Collectively, our results suggest that the Hsp90 inhibitor, pochonin D, could be an attractive antiviral therapeutic for treating HRV infection.

Serial Changes in Mannose-Binding Lectin in Patients with Sepsis / 결핵및호흡기질환

BACKGROUND: Mannose-binding lectin (MBL) deficiency leads to increased susceptibility to infection. We investigated whether serial changes in MBL levels are associated with the prognosis of patients diagnosed with septic shock, and correlated with cytokine levels. METHODS: We enrolled 131 patients with septic shock in the study. We analyzed the serum samples for MBL and cytokine levels at baseline and 7 days later. Samples on day 7 were available in 73 patients. RESULTS: We divided the patients with septic shock into four groups according to serum MBL levels ( < 1.3 µg/mL or ≥1.3 µg/mL) on days 1 and 7. Patients with low MBL levels on day 1 and high MBL levels on day 7 showed a favorable prognosis for 28-day survival (odds ratio, 1.96, 95% confidence interval, 1.10–2.87; p=0.087). The high MBL group on day 7 showed a significant decrease in monocyte chemoattractant protein 1, interleukin (IL)-1β, IL-6, IL-8, interferon-γ, and granulocyte macrophage colony-stimulating factor levels compared with the low MBL group on day 7. CONCLUSION: The increase in MBL levels of patients with septic shock may suggest a favorable prognosis and attenuate pro-inflammatory and anti-inflammatory responses.

Delta Neutrophil Index as an Early Marker for Distinguishing Myeloid from Childhood Acute Leukemia / 임상소아혈액종양

BACKGROUND: The accurate and early diagnosis of acute myeloid leukemia (AML) is important to choose proper treatment option depending on the risk stratification. The delta neutrophil index (DNI) is a relatively new blood marker that indicates the proportion of immature granulocytes in peripheral blood circulation. This study aimed to evaluate the diagnostic value of the DNI for detecting AML in the early phase of acute leukemia. METHODS: We retrospectively analyzed laboratory tests and bone marrow study results of 163 pediatric patients with acute leukemia admitted to the emergency department, who were diagnosed with acute leukemia. An automatic analyzer (ADVIA 2120 Hematology System; Siemens Healthcare Diagnostics, Forchheim, Germany) was used to measure the DNI in the peripheral blood of each patient. RESULTS: The mean DNI was significantly different between the AML (N=39) and non-AML (N=124) groups (P < 0.05), and the DNI was the only significant marker for predicting AML in patients with acute leukemia (odds ratio, 1.328; P < 0.05). The DNI more than 4.4% has the highest predictability for distinguishing the patients with AML from the patients with acute leukemia. The mean DNI of the acute promyelocytic leukemia (APL, N=8) group was statistically higher than that of the non-APL group (N=31, P=0.019), but the DNI was not significant in the univariate logistic regression analysis. CONCLUSION: The DNI might be a promising peripheral blood marker for predicting AML in the early work-up of patients with acute leukemia.

Dapsone modulates lipopolysaccharide-activated bone marrow cells by inducing cell death and down-regulating tumor necrosis factor-α production

Dapsone, an antibiotic, has been used to cure leprosy. It has been reported that dapsone has anti-inflammatory activity in hosts; however, the anti-inflammatory mechanism of dapsone has not been fully elucidated. The present study investigated the anti-inflammatory effects of dapsone on bone marrow cells (BMs), especially upon exposure to lipopolysaccharide (LPS). We treated BMs with LPS and dapsone, and the treated cells underwent cellular activity assay, flow cytometry analysis, cytokine production assessment, and reactive oxygen species assay. LPS distinctly activated BMs with several characteristics including high cellular activity, granulocyte changes, and tumor necrosis factor alpha (TNF-α) production increases. Interestingly, dapsone modulated the inflammatory cells, including granulocytes in LPS-treated BMs, by inducing cell death. While the percentage of Gr-1 positive cells was 57% in control cells, LPS increased that to 75%, and LPS plus dapsone decreased it to 64%. Furthermore, dapsone decreased the mitochondrial membrane potential of LPS-treated BMs. At a low concentration (25 µg/mL), dapsone significantly decreased the production of TNF-α in LPS-treated BMs by 54%. This study confirmed that dapsone has anti-inflammatory effects on LPS-mediated inflammation via modulation of the number and function of inflammatory cells, providing new and useful information for clinicians and researchers.

LRRC25 plays a key role in all-trans retinoic acid-induced granulocytic differentiation as a novel potential leukocyte differentiation antigen

Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.

Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea

The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5–100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.