Recent advancements in machine learning for bone marrow cell morphology analysis.

As machine learning progresses, techniques such as neural networks, decision trees, and support vector machines are being increasingly applied in the medical domain, especially for tasks involving large datasets, such as cell detection, recognition, classification, and visualization. Within the domain of bone marrow cell morphology analysis, deep learning offers substantial benefits due to its robustness, ability for automatic feature learning, and strong image characterization capabilities. Deep neural networks are a machine learning paradigm specifically tailored for image processing applications. Artificial intelligence serves as a potent tool in supporting the diagnostic process of clinical bone marrow cell morphology. Despite the potential of artificial intelligence to augment clinical diagnostics in this domain, manual analysis of bone marrow cell morphology remains the gold standard and an indispensable tool for identifying, diagnosing, and assessing the efficacy of hematologic disorders. However, the traditional manual approach is not without limitations and shortcomings, necessitating, the exploration of automated solutions for examining and analyzing bone marrow cytomorphology. This review provides a multidimensional account of six bone marrow cell morphology processes: automated bone marrow cell morphology detection, automated bone marrow cell morphology segmentation, automated bone marrow cell morphology identification, automated bone marrow cell morphology classification, automated bone marrow cell morphology enumeration, and automated bone marrow cell morphology diagnosis. Highlighting the attractiveness and potential of machine learning systems based on bone marrow cell morphology, the review synthesizes current research and recent advances in the application of machine learning in this field. The objective of this review is to offer recommendations to hematologists for selecting the most suitable machine learning algorithms to automate bone marrow cell morphology examinations, enabling swift and precise analysis of bone marrow cytopathic trends for early disease identification and diagnosis. Furthermore, the review endeavors to delineate potential future research avenues for machine learning-based applications in bone marrow cell morphology analysis.

Atorvastatin Treatment Significantly Increased the Concentration of Bone Marrow-Derived Mononuclear Cells and Transcutaneous Oxygen Pressure and Lowered the Pain Scale after Bone Marrow Cells Treatment in Patients with "No-Option" Critical Limb Ischaemia.

BACKGROUND: The present study investigated the outcomes and possible predictive factors of autologous bone marrow cells (BMCs) therapy in patients with "no-option" critical limb ischaemia (CLI). It was focused on exploring the clinical background and prior statin and renin-angiotensin system (RAS)-acting agents pharmacotherapy related to the therapeutic efficacy of BMCs treatment. METHODS: In the present study, we reviewed thirty-three patients (mean age 64.9 ± 10 years; 31 males) with advanced CLI after failed or impossible revascularisation, who were treated with 40 mL of autologous BMCs by local intramuscular application. Patients with limb salvage and wound healing (N = 22) were considered as responders to BMCs therapy, and patients with limb salvage and complete ischemic wound healing (N = 13) were defined as super-responders. Logistic regression models were used to screen and identify the prognostic factors, and a receiver operating characteristics (ROC) curve, a linear regression, and a survival curve were drawn to determine the predictive accuracy, the correlation between the candidate predictors, and the risk of major amputation. RESULTS: Based on the univariate regression analysis, baseline C-reactive protein (CRP) and transcutaneous oxygen pressure (TcPO2) values were identified as prognostic factors of the responders, while CRP value, ankle-brachial index (ABI), and bone marrow-derived mononuclear cells (BM-MNCs) concentration were identified as prognostic factors of the super-responders. An area under the ROC curve of 0.768 indicated good discrimination for CRP > 8.1 mg/L before transplantation as a predictive factor for negative clinical response. Linear regression analysis revealed a significant dependence between the levels of baseline CRP and the concentration of BM-MNCs in transplanted bone marrow. Patients taking atorvastatin before BMCs treatment (N = 22) had significantly improved TcPO2 and reduced pain scale after BMCs transplant, compared to the non-atorvastatin group. Statin treatment was associated with reduced risk for major amputation. However, the difference was not statistically significant. Statin use was also associated with a significantly higher concentration of BM-MNCs in the transplanted bone marrow compared to patients without statin treatment. Patients treated with RAS-acting agents (N = 20) had significantly reduced pain scale after BMCs transplant, compared to the non-RAS-acting agents group. Similar results, reduced pain scale and improved TcPO2, were achieved in patients treated with atorvastatin and RAS-acting agents (N = 17) before BMCs treatment. Results of the Spearman correlation showed a significant positive correlation between CLI regression, responders, and previous therapy before BMCs transplant with RAS-acting agents alone or with atorvastatin. CONCLUSIONS: CRP and TcPO2 were prognostic factors of the responders, while CRP value, ABI, and BM-MNCs concentration were identified as predictive factors of the super-responders. Atorvastatin treatment was associated with a significantly increased concentration of BM-MNCs in bone marrow concentrate and higher TcPO2 and lower pain scale after BMCs treatment in CLI patients. Similarly, reduced pain scales and improved TcPO2 were achieved in patients treated with atorvastatin and RAS-acting agents before BMCs treatment. Positive correlations between responders and previous treatment before BMCs transplant with RAS-acting agents alone or with atorvastatin were significant.

Biosynthesis and biocompatibility evaluation of zinc oxide nanoparticles prepared using Priestia megaterium bacteria.

The current study aimed to find an effective, simple, ecological, and nontoxic method for bacterial green synthesis of zinc oxide nanoparticles (ZnONPs) using the bacterial strain Priestia megaterium BASMA 2022 (OP572246). The biosynthesis was confirmed by the change in color of the cell-free supernatant added to the zinc nitrate from yellow to pale brown. The Priestia megaterium zinc oxide nanoparticles (Pm/ZnONPs) were characterized using UV-Vis spectroscopy, high-resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), and zeta potential. The Pm/ZnONPs characterization showed that they have a size ranging between 5.77 and 13.9 nm with a semi-sphere shape that is coated with a protein-carbohydrate complex. An EDX analysis of the Pm/ZnONPs revealed the presence of the shield matrix, which was composed of carbon, nitrogen, oxygen, chlorine, potassium, sodium, aluminum, sulfur, and zinc. The results of the FTIR analysis showed that the reduction and stabilization of the zinc salt solution were caused by the presence of O-H alcohols and phenols, O=C=O stretching of carbon dioxide, N=C=S stretching of isothiocyanate, and N-H bending of amine functional groups. The produced ZnONPs had good stability with a charge of - 16.2 mV, as evidenced by zeta potential analysis. The MTT assay revealed IC50 values of 8.42% and 200%, respectively, for the human A375 skin melanoma and human bone marrow 2M-302 cell lines. These findings revealed that the obtained Pm/ZnONPs have the biocompatibility to be applied in the pharmaceutical and biomedical sectors.

An Intelligent Attention-Based Transfer Learning Model for Accurate Differentiation of Bone Marrow Stains to Diagnose Hematological Disorder.

Bone marrow (BM) is an essential part of the hematopoietic system, which generates all of the body's blood cells and maintains the body's overall health and immune system. The classification of bone marrow cells is pivotal in both clinical and research settings because many hematological diseases, such as leukemia, myelodysplastic syndromes, and anemias, are diagnosed based on specific abnormalities in the number, type, or morphology of bone marrow cells. There is a requirement for developing a robust deep-learning algorithm to diagnose bone marrow cells to keep a close check on them. This study proposes a framework for categorizing bone marrow cells into seven classes. In the proposed framework, five transfer learning models-DenseNet121, EfficientNetB5, ResNet50, Xception, and MobileNetV2-are implemented into the bone marrow dataset to classify them into seven classes. The best-performing DenseNet121 model was fine-tuned by adding one batch-normalization layer, one dropout layer, and two dense layers. The proposed fine-tuned DenseNet121 model was optimized using several optimizers, such as AdaGrad, AdaDelta, Adamax, RMSprop, and SGD, along with different batch sizes of 16, 32, 64, and 128. The fine-tuned DenseNet121 model was integrated with an attention mechanism to improve its performance by allowing the model to focus on the most relevant features or regions of the image, which can be particularly beneficial in medical imaging, where certain regions might have critical diagnostic information. The proposed fine-tuned and integrated DenseNet121 achieved the highest accuracy, with a training success rate of 99.97% and a testing success rate of 97.01%. The key hyperparameters, such as batch size, number of epochs, and different optimizers, were all considered for optimizing these pre-trained models to select the best model. This study will help in medical research to effectively classify the BM cells to prevent diseases like leukemia.

Automated bone marrow cell classification through dual attention gates dense neural networks.

PURPOSE: The morphology of bone marrow cells is essential in identifying malignant hematological disorders. The automatic classification model of bone marrow cell morphology based on convolutional neural networks shows considerable promise in terms of diagnostic efficiency and accuracy. However, due to the lack of acceptable accuracy in bone marrow cell classification algorithms, automatic classification of bone marrow cells is now infrequently used in clinical facilities. To address the issue of precision, in this paper, we propose a Dual Attention Gates DenseNet (DAGDNet) to construct a novel efficient, and high-precision bone marrow cell classification model for enhancing the classification model's performance even further. METHODS: DAGDNet is constructed by embedding a novel dual attention gates (DAGs) mechanism in the architecture of DenseNet. DAGs are used to filter and highlight the position-related features in DenseNet to improve the precision and recall of neural network-based cell classifiers. We have constructed a dataset of bone marrow cell morphology from the First Affiliated Hospital of Chongqing Medical University, which mainly consists of leukemia samples, to train and test our proposed DAGDNet together with the bone marrow cell classification dataset. RESULTS: When evaluated on a multi-center dataset, experimental results show that our proposed DAGDNet outperforms image classification models such as DenseNet and ResNeXt in bone marrow cell classification performance. The mean precision of DAGDNet on the Munich Leukemia Laboratory dataset is 88.1%, achieving state-of-the-art performance while still maintaining high efficiency. CONCLUSION: Our data demonstrate that the DAGDNet can improve the efficacy of automatic bone marrow cell classification and can be exploited as an assisting diagnosis tool in clinical applications. Moreover, the DAGDNet is also an efficient model that can swiftly inspect a large number of bone marrow cells and offers the benefit of reducing the probability of an incorrect diagnosis.

Improved YOLOv7 Algorithm for Detecting Bone Marrow Cells.

The detection and classification of bone marrow (BM) cells is a critical cornerstone for hematology diagnosis. However, the low accuracy caused by few BM-cell data samples, subtle difference between classes, and small target size, pathologists still need to perform thousands of manual identifications daily. To address the above issues, we propose an improved BM-cell-detection algorithm in this paper, called YOLOv7-CTA. Firstly, to enhance the model's sensitivity to fine-grained features, we design a new module called CoTLAN in the backbone network to enable the model to perform long-term modeling between target feature information. Then, in order to cooperate with the CoTLAN module to pay more attention to the features in the area to be detected, we integrate the coordinate attention (CoordAtt) module between the CoTLAN modules to improve the model's attention to small target features. Finally, we cluster the target boxes of the BM cell dataset based on K-means++ to generate more suitable anchor boxes, which accelerates the convergence of the improved model. In addition, in order to solve the imbalance between positive and negative samples in BM-cell pictures, we use the Focal loss function to replace the multi-class cross entropy. Experimental results demonstrate that the best mean average precision (mAP) of the proposed model reaches 88.6%, which is an improvement of 12.9%, 8.3%, and 6.7% compared with that of the Faster R-CNN model, YOLOv5l model, and YOLOv7 model, respectively. This verifies the effectiveness and superiority of the YOLOv7-CTA model in BM-cell-detection tasks.

ATP-P2X7R-mediated microglia senescence aggravates retinal ganglion cell injury in chronic ocular hypertension.

BACKGROUND: Dysfunction of microglia during aging affects normal neuronal function and results in the occurrence of neurodegenerative diseases. Retinal microglial senescence attributes to retinal ganglion cell (RGC) death in glaucoma. This study aims to examine the role of ATP-P2X7R in the mediation of microglia senescence and glaucoma progression. METHODS: Forty-eight participants were enrolled, including 24 patients with primary open-angle glaucoma (POAG) and age-related cataract (ARC) and 24 patients with ARC only. We used ARC as the inclusion criteria because of the availability of aqueous humor (AH) before phacoemulsification. AH was collected and the adenosine triphosphate (ATP) concentration was measured by ATP Assay Kit. The chronic ocular hypertension (COH) mouse model was established by microbead occlusion. Microglia were ablated by feeding PLX5622 orally. Mouse bone marrow cells (BMCs) were prepared and infused into mice through the tail vein for the restoration of microglia function. Western blotting, qPCR and ELISA were performed to analyze protein and mRNA expression in the ocular tissue, respectively. Microglial phenotype and RGC survival were assessed by immunofluorescence. The mitochondrial membrane potential was measured using a JC-1 assay kit by flow cytometry. RESULTS: ATP concentrations in the AH were increased in older adults and patients with POAG. The expression of P2X7R was upregulated in the retinal tissues of mice with glaucoma, and functional enrichment analysis showed that P2X7R was closely related to cell aging. Through in vivo and in vitro approaches, we showed that pathological activation of ATP-P2X7R induced accelerated microglial senescence through impairing PTEN-induced kinase 1 (PINK1)-mediated mitophagy, which led to RGC damage. Additionally, we found that replacement of senescent microglia in COH model of old mice with BMCs from young mice reversed RGC damage. CONCLUSION: ATP-P2X7R induces microglia senescence by inhibiting PINK1-mediated mitophagy pathway. Specific inhibition of ATP-P2X7R may be a fundamental approach for targeted therapy of RGC injury in microglial aging-related glaucoma.

Harvest Quality, Nucleated Cell Dose and Clinical Outcomes in Bone Marrow Transplantation: A Retrospective Study.

Higher doses of infused nucleated cells (NCs) are associated with improved clinical outcomes in bone marrow transplantation (BMT) recipients. Most clinicians recommend infusing at least 2.0 × 108 NCs/kg. BMT clinicians request a target NC dose, but the harvested NC dose may be below the requested NC dose even before cell processing. We conducted this retrospective study to investigate the quality of bone marrow (BM) harvest and factors that influence infused NC doses at our institution. We also correlated infused NC doses with clinical outcomes. The study population included 347 BMT recipients (median age, 11 years; range, <1 to 75 years) at the University of Minnesota between 2009 and 2019. Underlying diagnoses mainly included 39% malignant and 61% nonmalignant diagnoses. Requested, harvested, and infused NC doses, as well as cell processing data, were obtained from the Cell Therapy Laboratory; clinical outcomes data were obtained from the University of Minnesota BMT Database. BM harvests were facilitated either by our institution (61%) or by the National Marrow Donor Program (39%). Associations of infused doses with baseline characteristics were assessed using the general Wilcoxon test/Pearson's correlation coefficient. The association of infused dose with neutrophil engraftment (absolute neutrophil count >500) by day 42, platelet engraftment (>20,000) by 6 months, acute graft-versus-host disease grade II-IV, and overall survival (OS) at 5 years were evaluated using regression and Kaplan-Meier curves. The median requested NC dose was 3.0 × 108/kg (range, 2 to 8 × 108/kg), and the median harvested and infused NC doses were 4.0 × 108/kg and 3.6 × 108/kg, respectively. Only 7% of donors had a harvested dose below the minimum requested dose. Moreover, the correlation between requested doses and harvested doses was adequate, with a harvested/requested dose ratio <.5 observed in only 5% of harvests. Additionally, the harvest volume and cell processing method were significantly correlated with the infused dose. Harvest volume exceeding the median of 948 mL was related to a significantly lower infused dose (P < .01). Moreover, hydroxyethyl starch (HES)/buffy coat processing (used to reduce RBCs with major ABO incompatibility) led to a significantly lower infused dose (P < .01). Donor age (median, 19 years; range, <1 to-70 years) and sex did not significantly influence the infused dose. Finally, the infused dose was significantly correlated with neutrophil and platelet engraftment (P < .05) but not with 5-year OS (P = .87) or aGVHD (P = .33). In our program's experience, BM harvesting is efficient and meets the requested minimum dose for 93% of recipients. Harvest volume and cell process play significant roles in determining the final infused dose. Minimizing harvest volume and cell processing could lead to increased infused dose and thus improved outcomes. Moreover, a higher infused dose leads to a better rate of neutrophil and platelet engraftment but not to improved OS, which may be linked to the sample size of our study.

Accumulation of Dystrophin-Positive Muscle Fibers and Improvement of Neuromuscular Junctions in mdx Mouse Muscles after Bone Marrow Transplantation under Different Conditions.

Duchenne muscular dystrophy (DMD) is a severe muscular disorder caused by mutations in the dystrophin gene. It leads to respiratory and cardiac failure and premature death at a young age. Although recent studies have greatly deepened the understanding of the primary and secondary pathogenetic mechanisms of DMD, an effective treatment remains elusive. In recent decades, stem cells have emerged as a novel therapeutic product for a variety of diseases. In this study, we investigated nonmyeloablative bone marrow cell (BMC) transplantation as a method of cell therapy for DMD in an mdx mouse model. By using BMC transplantation from GFP-positive mice, we confirmed that BMCs participate in the muscle restoration of mdx mice. We analyzed both syngeneic and allogeneic BMC transplantation under different conditions. Our data indicated that 3 Gy X-ray irradiation with subsequent BMC transplantation improved dystrophin synthesis and the structure of striated muscle fibers (SMFs) in mdx mice as well as decreasing the death rate of SMFs. In addition, we observed the normalization of neuromuscular junctions (NMJs) in mdx mice after nonmyeloablative BMC transplantation. In conclusion, we demonstrated that nonmyeloablative BMC transplantation could be considered a method for DMD treatment.

Matrix Metalloproteinase 1 as a Marker of Tonsil-Derived Mesenchymal Stem Cells to Assess Bone Marrow Cell Migration.

BACKGROUND: To achieve optimal bone marrow engraftment during bone marrow transplantation, migration of donor bone marrow cells (BMCs) toward the recipient's bone marrow is critical. Despite the enhanced engraftment of BMCs by co-administration of mesenchymal stem cells (MSCs), the efficiency can be variable depending on MSC donor. The purpose of this study is to examine the functional heterogeneity of tonsil-derived MSCs (TMSCs) and to identify a marker to evaluate efficacy for the enhancement of BMC migration. METHODS: To examine the donor-to-donor variation of TMSCs in potentiating BMC migration, we isolated TMSCs from 25 independent donors. Transcriptome of TMSCs and proteome of conditioned medium derived from TMSC were analyzed. RESULTS: Enhanced BMC migration by conditioned medium derived from TMSCs was variable depending on TMSC donor. The TMSCs derived from 25 donors showed distinct expression profiles compared with other cells, including fibroblasts, adipose-derived MSCs and bone marrow-derived MSCs. TMSCs were distributed in two categories: high- and low-efficacy groups for potentiating BMC migration. Transcriptome analysis of TMSCs and proteome profiles of conditioned medium derived from TMSCs revealed higher expression and secretion of matrix metalloproteinase (MMP) 1 in the high-efficacy group. MMP1 knockdown in TMSCs abrogated the supportive efficacy of conditioned medium derived from TMSC cultures in BMC migration. CONCLUSION: These data suggest that secreted MMP1 can be used as a marker to evaluate the efficacy of TMSCs in enhancing BMC migration. Furthermore, the strategy of analyzing transcriptomes and proteomes of the MSCs may be useful to set the standard for donor variation.

Successful Stem Cell Mobilization and CD34+ Cell Collection in a Poor Mobilizer: A Case Report Utilizing a Combination of Recombinant Growth Colony Stimulating Factor, Recombinant Human Growth Factor, and Plerixafor.

Diffuse large B-cell lymphoma is the most prevalent form of non-Hodgkin's lymphoma that is usually treated with chemoimmunotherapy. If the disease proves refractory or recurrent, the primary treatment approach involves high-dose chemotherapy with bone marrow transplantation. The collection of peripheral blood stem cells before transplantation plays a vital role in the treatment process, necessitating the mobilization of blood stem cells from the bone marrow to the peripheral blood. Despite using standard methods such as granulocyte colony-stimulating factor (G-CSF), chemotherapy, and plerixafor, some patients cannot collect an optimal count of CD34+ cells for transplantation. Managing these patients with poor mobilization poses significant challenges. In this article, we present a case of a poor mobilizer patient who achieved prosperous mobilization by using recombinant human G-CSF, recombinant human growth hormone, and plerixafor.

[Characteristics of Bone Marrow Cell Morphology and Immunophenotype in Patients with NHL Leukemia and Evaluation of NHL Bone Marrow Invasion].

OBJECTIVE: To observe the evaluation value of characteristics of bone marrow cell morphology and immunophenotype in patients with non-Hodgkin lymphoma (NHL) leukemia on the bone marrow invasion of NHL. METHODS: The clinical data of 104 patients with NHL treated in the hospital from March 2016 to March 2021 were retrospectively analyzed, the characteristics of bone marrow smear morphology were recorded, and the analysis of bone marrow immunophenotype was performed, the evaluation value of bone marrow cell morphology and immunophenotype on NHL bone marrow invasion was analyzed. RESULTS: One hundred and four patients with NHL leukemia were found to have increased lymphoma cells by examination of bone marrow cell morphology, including 57 cases of B-cell type, 39 cases of T-cell type and 8 cases of NK/T cell type. The characteristics of bone marrow cell morphology were as follows: the cell body was large, irregular or round like in shape, the cytoplasm was much and mostly stained blue, a few cells could see a few granules, the nucleus was round or round like, some were twisted, the chromatin was thick and nucleolus was different, of which the T-cell type lymphoma cells had strong heteromorphism and more obvious nucleolus. B-cell type mainly expressed CD19, HLA-DR and CD20, and the positive rate was ≥70%. T-cell type mainly expressed CD7, HLA-DR and CD38, and the positive rate was ≥40%. NK/T cell types mainly expressed CD56 and CD161, and the positive rates was ≥50%. Compared the proportion of lymphoma cells between bone marrow smear and immunophenotype examination, there was no statistical significant difference (P>0.05). CONCLUSION: The characteristics of bone marrow cell morphology and immunophenotype have certain application value in the evaluation of bone marrow invasion in patients with NHL leukemia, both can complement each other and provide a feasible mean for the effective evaluation of bone marrow invasion in patients with NHL leukemia.

Diagnostic value of bone marrow cell morphology in visceral leishmaniasis-associated hemophagocytic syndrome: Two case reports.

BACKGROUND: Visceral leishmaniasis related-hemophagocytic lymphohistiocytosis (VL-HLH) is a hemophagocytic syndrome caused by Leishmania infection. VL-HLH is rare, especially in nonendemic areas where the disease is severe, and mortality rates are high. The key to diagnosing VL-HLH is to find the pathogen; therefore, the Leishmania must be accurately identified for timely clinical treatment. CASE SUMMARY: We retrospectively analyzed the clinical data, laboratory examination results, and bone marrow cell morphology of two children with VL-HLH diagnosed via bone marrow cell morphology at Kunming Children's Hospital of Yunnan, China. Both cases suspected of having malignant tumors at other hospitals and who were unresponsive to treatment were transferred to Kunming Children's Hospital. They are Han Chinese girls, one was 2 years old and the other one is 9 mo old. They had repeated fevers, pancytopenia, hepatosplenomegaly, hypertriglyceridemia, and hypofibrinogenemia over a long period and met the HLH-2004 criteria. Their HLH genetic test results were negative. Both children underwent chemotherapy as per the HLH-2004 chemotherapy regimen, but it was ineffective and accompanied by serious infections. We found Leishmania amastigotes in their bone marrow via morphological examination of their bone marrow cells, which showed hemophagocytic cells; thus, the children were diagnosed with VL-HLH. After being transferred to a specialty hospital for treatment, the condition was well-controlled. CONCLUSION: Morphological examination of bone marrow cells plays an important role in diagnosing VL-HLH. When clinically diagnosing secondary HLH, VL-HLH should be considered in addition to common pathogens, especially in patients for whom HLH-2004 chemotherapy regimens are ineffective. For infants and young children, bone marrow cytology examinations should be performed several times and as early as possible to find the pathogens to reduce potential misdiagnoses.

[Morphological Characteristics of Bone Marrow Cells in Patients with EB Virus Infection].

OBJECTIVE: Review and analyze the characteristics of bone marrow cell morphology in patients with Epstein-Barr virus (EBV) infection, and explore the diagnostic value of bone marrow cell morphology for the early identification of EBV infection. METHODS: A total of 33 patients with EBV-DNA positive detection in the First Affiliated Hospital of Guangxi Medical University from January 2018 to May 2021 were collected as the research objects. Bone marrow cell morphology and peripheral blood cell analysis were performed, and the significance in disease diagnosis was analyzed by statistical methods. RESULTS: The sampling satisfaction of 33 patients with EBV infection was 100%. In the clinical diagnosis of all cases, 7 cases were IM, 17 cases were EBV-HLH, 3 cases were lymphoma, 2 cases were EBV-associated lymphoid hyperplasia, and 4 cases were not diagnosed. Among them, 31 patients had active bone marrow hyperplasia or above, 26 patients had active granulocytic hyperplasia or above, 21 patients had active erythroid hyperplasia or above, and 17 cases of megakaryocyte production platelet function decreased. The abnormal components of bone marrow mainly indude atypical lymphocyte cells (33 cases), hemophagocytic cells (22 cases), abnormal histiocyte (10 cases). CONCLUSION: According to the proliferation of granulocytes, erythrocytes and megakaryocytes in the bone marrow, and the emergence of abnormal components such as atypical lymphocytes, hemophagocyte, abnormal histiocyte. Bone marrow cell morphological examination can indicate the possibility of EBV infection, which is certain diagnostic value for early identification of EBV infection.

[Radioprotective effects of gallic acid on bone marrow cells in mice].

OBJECTIVE: To investigate the radioprotective effect of gallic acid(GA) on mouse bone marrow cells. METHODS: Healthy male ICR mice were randomly divided into saline control group, GA control group, X-ray irradiation group and GA protection group, with 10 mice in each group. X-ray irradiation group and normal saline control group were given 0.01 mL/g normal saline gavage, GA control group and GA protection group were given 200 mg/kg GA(20 mg/mL) gavage once a day for 14 consecutive days. On the 15 th day, 4 X-ray irradiation groups and 4 GA protection groups were given one-time X-ray irradiation to the whole body of the mice, and the absorbed doses were 1.0, 2.0, 3.0 and 4.0 Gy, respectively. The saline control group and the GA control group were not irradiated. After irradiation, detected the whole blood catalase(CAT), superoxide dismutase(SOD), malondialdehyde(MDA) and micronucleus frequency of polychromatic erythrocyte in bone marrow(MN-PCE), and use flow cytometry to detect bone marrow cell cycle, early apoptosis rate and late apoptosis rate. RESULTS: The CAT activities in the serum of mice in the 1.0, 2.0, 3.0 and 4.0 Gy GA protection groups were 2.13, 1.74, 1.49 and 1.15 U/mL, respectively, which were significantly increased compared with the corresponding X-ray irradiation group(P<0.01). SOD activities were 184.69, 156.92, 139.17 and 107.15 U/mL, which were significantly increased compared with the corresponding X-ray irradiation group(P<0.01). The contents of MDA were 3.92, 4.20, 6.32 and 9.31 nmol/mL, which were significantly lower than those of the corresponding X-ray irradiation group(P<0.05, P<0.01). The bone marrow MN-PCE rate of the mice in the 1.0, 2.0, 3.0 and 4.0 Gy GA protection groups were 4.35‰, 8.00‰, 12.90‰ and 3.80‰, respectively, which were significantly lower than those in the corresponding X-ray irradiation group(P<0.01). The proportions of G_0/G_1 phase cells of bone marrow cells in the 1.0, 2.0, 3.0 and 4.0 Gy GA protection group were 81.00%, 86.28%, 92.04% and 93.15%, respectively, which were significantly reduced compared with the corresponding X-ray irradiation group(P<0.01). The proportions of G_2/M phase cells were 4.51%, 3.05%, 2.35% and 1.81%, which were significantly increased compared with the corresponding X-ray irradiation group(P<0.05, P<0.01). The proportions of S phase cells were 15.32%, 11.36%, 5.96% and 4.92%, which were significantly increased compared with the corresponding X-ray irradiation group(P<0.01). The early apoptosis rate of bone marrow cells of mice in the 1.0, 2.0, 3.0 and 4.0 Gy GA protection groups were 3.32%, 8.96%, 12.11% and 2.26%, respectively, which were significantly reduced compared with the corresponding X-ray irradiation group(P<0.01). The late apoptosis rates of bone marrow cells were 7.21%, 11.73%, 17.11% and 19.36%, which were significantly reduced compared with the corresponding X-ray irradiation group(P<0.05, P<0.01). CONCLUSION: GA can reduce the oxidative damage, DNA damage and bone marrow cell cycle arrest of the bone marrow cells of radiation-damaged mice, inhibit the apoptosis of bone marrow cells, and have radioprotective effects on the bone marrow cells of mice.

Mouse Bone Marrow Cell Isolation and Macrophage Differentiation.

The rapid increase in the incidence of obesity contributes to a parallel increase in nonalcoholic steatohepatitis (NASH). Monocyte-derived macrophages, recruited from the bone marrow to the liver, promote NASH-related inflammation and fibrosis. In addition, adipose tissue macrophages (ATMs) release pro-inflammatory cytokines (PICs) which stimulate adipose tissue lipolysis liberating free fatty acids (FFAs) that can accumulate in the liver as triglycerides (TGs), thereby inducing steatosis. As such, bone marrow-derived macrophages (BMDMs) function as an essential tool to study the pathogenesis of NASH. BMDMs are primary bone marrow-derived cells which are differentiated into macrophages in vitro in the presence of growth factors. Macrophage colony-stimulating factor (M-CSF) is required for the proliferation and differentiation of committed myeloid progenitors into cells of the macrophage/monocyte lineage. Here, we describe a protocol for the isolation of mouse bone marrow cells and subsequent macrophage differentiation in which bone marrow cells are cultured in the presence of M-CSF, supplemented either by conditioned medium from L929 cells or in purified form. The efficiency of the differentiation is confirmed by immunofluorescent staining of macrophage surface antigen F4/80. The BMDMs serve as an excellent ex vivo model for a variety of studies, including hepatocyte-macrophage and adipocyte-macrophage cross-talk regulating NASH.

Comparing the efficacy of adipose-derived and bone marrow-derived cells in a rat model of posterolateral lumbar fusion.

Although bone marrow-derived mesenchymal stem cells (BMCs) have been widely used in spinal fusion procedures, adipose-derived stem cells (ASCs) offer a number of advantages as an alternative clinical cell source. This study directly compares the efficacy of ASCs and BMCs from the same donor animals to achieve successful fusion when combined with a clinical-grade bone graft substitute in a rat lumbar fusion model. ASCs and BMCs were isolated from the same Lewis donor rats and grown to passage 2 (P2). Single-level bilateral posterolateral intertransverse process lumbar fusion surgery was performed on syngeneic rats divided into three experimental groups: clinical-grade bone graft substitute alone (CBGS); CBGS+ rat ASCs (rASC); and, CBGS+ rat BMCs (rBMC). Eight weeks postoperatively, fusion was evaluated via micro-CT, manual palpation and histology. In vitro analysis of the osteogenic capacity of rBMCs and rASCs was also performed. Results indicated that the average fusion volume in the rASC group was the largest and was significantly larger than the CBGS group. Although the rASC group displayed the highest fusion rates via micro-CT and manual palpation, this difference was not statistically significant. Cell-seeded grafts showed more histological bone formation than cell-free grafts. P2 rASCs and rBMCs displayed similar in vitro osteogenic differentiation capacities. Overall, this study showed that, when combined with a clinical-grade bone graft substitute in a rat model, rASCs cells yielded the largest fusion masses and comparable fusion results to rBMCs. These results add to growing evidence that ASCs provide an attractive alternative to BMCs for spinal fusion procedures.

Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time.

Chicken bone marrow-derived macrophages (BMMΦ) and dendritic cells (BMDC) are utilized as models to study the mononuclear phagocytic system (MPS). A widely used method to generate macrophages and DC in vitro is to culture bone marrow cells in the presence of colony-stimulating factor-1 (CSF1) to differentiate BMMΦ and granulocyte-macrophage-CSF (GM-CSF, CSF2) and interleukin-4 (IL-4) to differentiate BMDC, while CSF2 alone can lead to the development of granulocyte-macrophage-CSF-derived DC (GMDC). However, in chickens, the MPS cell lineages and their functions represented by these cultures are poorly understood. Here, we decipher the phenotypical, functional and transcriptional differences between chicken BMMΦ and BMDC along with examining differences in DC cultures grown in the absence of IL-4 on days 2, 4, 6 and 8 of culture. BMMΦ cultures develop into a morphologically homogenous cell population in contrast to the BMDC and GMDC cultures, which produce morphologically heterogeneous cell cultures. At a phenotypical level, all cultures contained similar cell percentages and expression levels of MHCII, CD11c and CSF1R-transgene, whilst MRC1L-B expression decreased over time in BMMΦ. All cultures were efficiently able to uptake 0.5 µm beads, but poorly phagocytosed 1 µm beads. Little difference was observed in the kinetics of phagosomal acidification across the cultures on each day of analysis. Temporal transcriptomic analysis indicated that all cultures expressed high levels of CSF3R, MERTK, SEPP1, SPI1 and TLR4, genes associated with macrophages in mammals. In contrast, low levels of FLT3, XCR1 and CAMD1, genes associated with DC, were expressed at day 2 in BMDC and GMDC after which expression levels decreased. Collectively, chicken CSF2 + IL-4- and CSF2-dependent BM cultures represent cells of the macrophage lineage rather than inducing conventional DC.

A case of epithelioid hemangioendothelioma diagnosed by bone marrow cell morphology / 中华检验医学杂志

Epithelioid hemangioendothelioma (EHE) is a rare malignant vascular tumor. Its malignancy is between benign hemangioma and highly malignant angiosarcoma. It originates from vascular endothelial cells or pre-endothelial cells. It is characterized by the proliferation of vascular endothelial cells with a skin-like or histiocyte-like appearance. The incidence of EHE is less than 1% in all vascular tumors, and it can occur in multiple parts of the body, most often in the liver, followed by simultaneous involvement of the liver and lung, the lung alone, and the bone alone. At present, there is no report of epithelioid hemangioendothelioma diagnosed by bone marrow cell morphological examination in China. In this case, abnormal cells were found through bone marrow cell morphological examination, which guided the direction of further diagnosis and treatment. And finally the patient was diagnosed as epithelioid hemangioendothelioma. The bone marrow cell morphological examination can provided an important basis for clinical diagnosis and treatment. Epithelioid hemangioendothelioma needs to be differentiated from a variety of benign and malignant angiogenic tumors, especially other types of epithelioid angiogenic tumors. At present, it has been found that the disease has characters of cytogenetic and molecular biological abnormalities. Combined with histopathological morphology and immunohistochemical examination, we can make the diagnosis and differential diagnosis.