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Purpose: Diagnosis of myelodysplastic syndrome (MDS) primarily relies on the detection of morphological dysplasia in bone marrow. It is subjective and many studies have reported lack of interobserver agreement in reporting. Biopsy is preferred specimen for megakaryocyte assessment. We studied 43 bone marrow biopsies from 40 suspected MDS patient having persistent undiagnosed cytopenia. Utility of immunohistochemistry (IHC) with CD61 and p53 in detecting low-grade MDS was analyzed over routine morphology. Method and Results: Total number of megakaryocytes and number of dysplastic megakaryocytes seen on CD61 IHC was significantly higher than that on H and E stain (P value < 0.05) Out of total 43 biopsies, 13 [30.2%] cases showed dysplastic megakaryocytes that were confirmed by interobserver agreement after IHC. From 30 cases with no significant dysplasia on morphology, 21/43 [48.8%] cases showed >10% dysplastic megakaryocytes on CD61 (P value 0.0001). Nine cases showed no significant dysmegakaryopoiesis with either H and E or CD61 IHC. Fourteen cases could meet higher cut off (30%) of dysmegakaryopoiesis with CD 61 IHC. Out of total 34 cases showing significant dysplasia 7 cases (20.6%) showed positivity for p53 on IHC, which is little less than that reported in low-grade MDS. Conclusion: CD61 IHC is helpful in making correct diagnosis of MDS in cases with minimal dysplasia and should be performed before excluding possibility of MDS on morphology in a patient with undiagnosed cytopenia. IHC is cost effective tool for MDS diagnosis in developing world where access to extensive flow cytometery and molecular testing is limited.
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【Objective】 To optimize the existing spin-EB method and promote human induced pluripotent stem cells (hiPSCs) differentiate into megakaryocytes (MKs). 【Methods】 In this study, the initial inoculation amount of hiPSCs was increased from 3 500 cells/well to 8 000 cells/well, and the size of EB was increased. By observing the generation time of EB- hematopoietic cells during differentiation, and detecting the proliferation of CD34+ hematopoietic progenitor cells and CD41+ MKs in different stages, it was studied whether the optimized scheme could promote the differentiation of hiPSCs into hematopoietic progenitor cells(HPCs) and MKs. 【Results】 By increasing the initial inoculation amount of hiPSCs and the size of EB, the differentiation of hiPSCs into HPCs and MKs and the cell production efficiency can be promoted. 【Conclusion】 Our research describes an optimized and repeatable differentiation method, which can produce hematopoietic progenitor cells and mature MKs from hiPSCs in a relatively short time with higher yield. It is of great clinical significance and broad scientific research prospect to continuously optimize the culture scheme of hiPSCs differentiation to produce MKs and platelets in vitro, and to promote large-scale platelet generation in vitro in transfusion medicine.
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Objective:To study the different factors affecting platelet production post transplantation of hematopoietic stem cells (HSCs) isolated from different sources in order to explore novel options for treating platelet depletion following HSCs transplantation.Methods:HSCs and their downstream derivatives including myeloid and lymphoid cells (i.e., collective of mononuclear cells (MNCs)), were isolated from E14.5 fetal liver (FL) and bone marrow (BM) of 8-week-old mice by Ficoll separation technique. These cells were subsequently transplanted into the tibia bone marrow cavity of recipient mice post lethal myeloablative treatment in order to construct the FL-MNCs and BM-MNCs transplantation mouse model. Routine blood indices were examined in these recipient mice. The chimeric rate of donor cells in recipient peripheral blood cells were determined by flow cytometry. Different groups of cells involved in platelet reconstruction were analyzed. CD41 +megakaryocytes were sorted from fetal liver or bone marrow using magnetic beads, which were then induced to differentiate into platelets in an in vitro assay . Quantitative RT-PCR was used to detect the expression of platelet-related genes in CD41 +megakaryocytes from the two sources. Results:Both the FL-MNCs and the BM-MNCs transplantation groups resumed normal hematopoiesis at the 4th week after transplantation, and the blood cells of the recipient mice were largely replaced by the donor cells. Compared with the mice transplanted with BM-MNCs, the platelet level of mice transplanted with FL-MNCs recovered faster and were maintained at a higher level. At week 4, the PLT level of the FL-MNCs group was (1.45±0.37)×10 12/L, and of the BM-MNCs group was (1.22±0.24)×10 12/L, P<0.05. The FL-MNCs contain a higher proportion of hematopoietic stem cells (Lin -Sca-1 +c-Kit +)(7.60%±1.40%) compared to the BM-MNCs (1.10%±0.46%), P<0.01; the proportion of the megakaryocyte progenitor cells (Lin -Sca-1 -c-Kit +CD41 +CD150 +) and mature megakaryocyte cells (CD41 +CD42b +), also differ significantly between the FL-MNCs (3.05%±0.22%, 1.60%±0.06%, respectively) and the BM-MNCs (0.15%±0.02%, 0.87%±0.11%, respectively) groups, both P<0.01. In vitro functional studies showed that FL-MNCs-CD41 +megakaryocytes could produce proplatelet-like cells more quickly after induction, with proplatelet-like cells formation on day 3 and significant platelet-like particle formation on day 5, in contrast to bone marrow-derived BM-MNCs-CD41 +megakaryocytes that failed to form proplatelet-like cell on day 5. In addition, FL-MNCs-CD41 +cells expressed higher levels of platelet-related genes, Mpl (3.25-fold), Fog1 (3-fold), and Gata1 (1.5-fold) ( P<0.05). Conclusion:Compared with the BM-MNCs group, the FL-MNCs transplantation group appears to have a more efficient platelet implantation effect in the HSCs transplantation recipient in vivo , as well as a higher platelet differentiation rate in vitro. This might be related to a higher proportion of megakaryocytes and higher expression levels of genes such as Mpl, Fog1, and Gata1 that could be important for platelet formation in FL-MNCs-CD41 +cells. Further exploration of the specific functions of these genes and the characteristics of the different proportions of the donor cells will provide valuable clues for the future treatment of platelets reconstitution after HSCs transplantation clinically.
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Platelet transfusion is the main clinical treatment of thrombocytopenia. However, due to the difficulty of platelet collection, high cost of use and limited number of blood donors, the development of platelet treatment is greatly limited. Therefore, the research on thrombogenesis in vitro has attracted more attention at home and abroad. Platelet production in vitro has the advantages of donor-independence, platelet antigen free and low risk of alloimmunity. At present, the efficiency of producing functional platelets in vitro is low, and there is still a big gap to achieve the ultimate goal of producing a large number of functional platelets in vitro. This paper reviews the research progress of megakaryocyte / platelet production in vitro, focuses on the in vitro production potential of megakaryocyte / platelet, and summarizes the current platelet culture systems in vitro based on human pluripotent stem cells, embryonic stem cells and adipose stem cells. The contradictions and difficulties of platelet production in vitro were also discussed to provide theoretical support for further research.
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Thrombocytopenia (platelet count less than 150,000/µl) is commonly encountered in routine hematological investigations. Here we present a study done to understand the prevalence of various conditions leading to thrombocytopenia, referred for bone marrow examination. It is a retrospective study done on 100 cases of thrombocytopenia referred for bone marrow examination in a tertiary care hospital from January 2016 to October 2016. The commonest cause of thrombocytopenia for which bone marrow was sought came out to be megaloblastic anemia followed by acute leukemia and aplastic anemia. Aim: Calculate the prevalence of various conditions causing thrombocytopenia, in cases referred for bone marrow examination, and Understand the various megakaryocytic alterations in hematological disorders presenting with thrombocytopenia due to different mechanisms. Methods: A retrospective study was done on 100 patients of thrombocytopenia referred for bone-marrow aspiration in a tertiary care hospital catering to both rural and urban population from January 2016 to October 2016. All cases of thrombocytopenia (platelet count less than 1,50,000/µl) diagnosed on hematology analyzer and later confirmed by peripheral blood film examination, referred for bone marrow examination for various reasons were included in this study. Stained bone-marrow aspirate smears were examined. Records regarding the clinical indication for the procedure, peripheral blood smear reports, blood counts and significant findings on bone-marrow aspiration smears were retrieved. The role of bone-marrow aspiration in the diagnosis of hematological and non- hematological disorders was reviewed in the study. Results: The commonest cause of thrombocytopenia for which bone marrow examination was sought was megaloblastic anemia(76%), followed by acute leukemia(7%), aplastic anemia(5%), myelodysplastic syndrome (4%) which was followed by ITP(3%), and one case each of gelatinous marrow transformation, malaria and NHL spillover. Conclusion: Further studies on the evaluation of megakaryocytic alteration and their contribution to thrombocytopenia can provide growing knowledge to the pathogenesis of numerous hematopoietic disorders that may identify broader clinical applications of the newer strategies to regulate platelet count and functioning.
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Objective@#To investigate the effect of recombinant human thrombopoietin (rhTPO) on thrombocytopenia (TCP) induced by endotoxin lipopolysaccharide (LPS) in mice.@*Methods@#Sixty male C57BL/6 mice were divided into normal saline (NS) control group (NS group), sepsis-induced TCP model group (LPS group) and rhTPO treatment group (LPS+rhTPO group) by random number table with 20 mice in each group. Sepsis-induced TCP model was reproduced by one intraperitoneal injection of LPS 30 mg/kg, and the mice in NS group were given the same amount of NS. In LPS+rhTPO group, 2.7 kU/kg rhTPO was subcutaneously injected into mice immediately after intraperitoneal injection of LPS, once every 24 hours. The mice in NS group and LPS group were injected subcutaneously with the same amount of NS. The observation period of each group lasted for 72 hours. The inner canthus blood was harvested before and every 24 hours after modeling, and the platelet count (PLT) was measured by animal blood cell counter. The eyeball blood of mice was harvested at 72 hours after modeling, and the proportion of CD61+CD62p+ cells in platelet-rich plasma was detected by flow cytometry, by which the platelet activation was reflected. Lung and spleen tissues of mice were harvested, and the positive expression of CD41 was determined by immunohistochemistry, by which the platelet sequestration in organs was reflected. Bone marrow cells from unilateral femur of mice were harvested, and the proportion of CD41+CD61+ cells was determined by flow cytometry to reflect the proliferation of bone marrow megakaryocytes.@*Results@#There was no significant difference in PLT among the groups before modeling. With the extension of the time after modeling, PLT in LPS group was decreased continuously, and increased slightly at 72 hours, but it was still significantly lower than that in NS group (×109/L: 308.60±21.70 vs. 1 152.72±50.27, P < 0.05); PLT in LPS+rhTPO group was increased continuously with the extension of modeling time, and it was significantly higher at 72 hours than that in LPS group (×109/L: 926.78±48.85 vs. 308.60±21.70, P < 0.05). At 72 hours after modeling, the proportion of CD61+CD62p+ cells in platelet-rich plasma of LPS group was significantly higher than that of NS group [(25.07±2.55)% vs. (4.17±0.38)%, P < 0.05], while the value in LPS+rhTPO group was significantly lower than that of LPS group [(15.92±1.26)% vs. (25.07±2.55)%, P < 0.05]. The proportion of CD41+CD61+ cells in bone marrow megakaryocytes of LPS group was significantly higher than that of NS group [(11.84±0.80)% vs. (3.60±0.42)%, P < 0.05], and the proportion of CD41+CD61+ cells in LPS+rhTPO group was significantly higher than that in LPS group [(30.96±2.49)% vs. (11.84±0.80)%, P < 0.05]. Immunohistochemistry showed that the positive expressions of CD41 in lung and spleen tissues of LPS group increased significantly than NS group [A value: 828.94±119.30 vs. 447.09±16.19 in lung tissue, (280.15±16.71)×103 vs. (0.65±0.26)×103 in spleen tissue, both P < 0.05], while the positive expressions of CD41 in lung and spleen tissues of LPS+rhTPO group decreased significantly than LPS group [A value: 542.78±2.95 vs. 828.94±119.30 in lung tissue, (129.40±13.49)×103 vs. (280.15±16.71)×103 in spleen tissue, both P < 0.05].@*Conclusion@#The rhTPO in endotoxin-induced TCP may stimulate the proliferation of bone marrow megakaryocytes, inhibit platelet activation and affect platelet sequestration in organs, so as to increase platelet levels.
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Objective To investigate the effect of recombinant human thrombopoietin (rhTPO) on thrombocytopenia (TCP) induced by endotoxin lipopolysaccharide (LPS) in mice. Methods Sixty male C57BL/6 mice were divided into normal saline (NS) control group (NS group), sepsis-induced TCP model group (LPS group) and rhTPO treatment group (LPS+rhTPO group) by random number table with 20 mice in each group. Sepsis-induced TCP model was reproduced by one intraperitoneal injection of LPS 30 mg/kg, and the mice in NS group were given the same amount of NS. In LPS+rhTPO group, 2.7 kU/kg rhTPO was subcutaneously injected into mice immediately after intraperitoneal injection of LPS, once every 24 hours. The mice in NS group and LPS group were injected subcutaneously with the same amount of NS. The observation period of each group lasted for 72 hours. The inner canthus blood was harvested before and every 24 hours after modeling, and the platelet count (PLT) was measured by animal blood cell counter. The eyeball blood of mice was harvested at 72 hours after modeling, and the proportion of CD61+CD62p+ cells in platelet-rich plasma was detected by flow cytometry, by which the platelet activation was reflected. Lung and spleen tissues of mice were harvested, and the positive expression of CD41 was determined by immunohistochemistry, by which the platelet sequestration in organs was reflected. Bone marrow cells from unilateral femur of mice were harvested, and the proportion of CD41+CD61+ cells was determined by flow cytometry to reflect the proliferation of bone marrow megakaryocytes. Results There was no significant difference in PLT among the groups before modeling. With the extension of the time after modeling, PLT in LPS group was decreased continuously, and increased slightly at 72 hours, but it was still significantly lower than that in NS group (×109/L: 308.60±21.70 vs. 1 152.72±50.27, P < 0.05); PLT in LPS+rhTPO group was increased continuously with the extension of modeling time, and it was significantly higher at 72 hours than that in LPS group (×109/L: 926.78±48.85 vs. 308.60±21.70, P < 0.05). At 72 hours after modeling, the proportion of CD61+CD62p+ cells in platelet-rich plasma of LPS group was significantly higher than that of NS group [(25.07±2.55)% vs. (4.17±0.38)%, P < 0.05], while the value in LPS+rhTPO group was significantly lower than that of LPS group [(15.92±1.26)% vs. (25.07±2.55)%, P < 0.05]. The proportion of CD41+CD61+ cells in bone marrow megakaryocytes of LPS group was significantly higher than that of NS group [(11.84±0.80)% vs. (3.60±0.42)%, P < 0.05], and the proportion of CD41+CD61+ cells in LPS+rhTPO group was significantly higher than that in LPS group [(30.96±2.49)% vs. (11.84±0.80)%, P < 0.05]. Immunohistochemistry showed that the positive expressions of CD41 in lung and spleen tissues of LPS group increased significantly than NS group [A value: 828.94±119.30 vs. 447.09±16.19 in lung tissue, (280.15±16.71)×103 vs. (0.65±0.26)×103 in spleen tissue, both P < 0.05], while the positive expressions of CD41 in lung and spleen tissues of LPS+rhTPO group decreased significantly than LPS group [A value: 542.78±2.95 vs. 828.94±119.30 in lung tissue, (129.40±13.49)×103 vs. (280.15±16.71)×103 in spleen tissue, both P < 0.05]. Conclusion The rhTPO in endotoxin-induced TCP may stimulate the proliferation of bone marrow megakaryocytes, inhibit platelet activation and affect platelet sequestration in organs, so as to increase platelet levels.
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Objective@#To analyze the clinical value of megakaryocytes in the diagnosis and treatment of children with immune thrombocytopenic purpura(ITP).@*Methods@#From June 2014 to January 2018, the clinical data of 110 children with ITP diagnosed and treated in Zhucheng People's Hospital Affiliated to Weifang Medical College were analyzed and followed up for more than 1 year.The children were divided into two groups according to whether the duration of the disease was morethan 12 months(chronic group and non-chronic group). Gender, age, initial course of disease, platelet count, lymphocyte count, megakaryocyte count, white blood cell count, and initial treatment regimen were analyzed and compared between the two groups.Multivariate analysis was used to analyze the independent influencing factors of chronic ITP.The clinical value of the initial diagnosis and lymphocyte counts in evaluation of the effects of chronic ITP and initial treatment were analyzed.The clinical value of megakaryocyte in the assessment of initial treatment was analyzed.@*Results@#The initial course of disease[(5.8±2.26)d]and megakaryocyte count[(210.28±98.67)/piece] in the chronic groupwere higher than those in the non-chronic group[(3.57±2.05)d, (165.26±78.35)/piece], and the lymphocyte count[(2.87±0.90)×109/L] in the chronic groupwas lower than that in the non-chronic group[(3.66±1.12)×109/L], the differences were statistically significant(t=4.824, 2.299, 3.545, all P<0.05). Megakaryocyte count was not an independent factor of chronic ITP(P>0.05). The initial course of disease was a risk factor for chronic ITP(OR=3.826), while lymphocyte count was a protective factor(OR=0.471). The initial course of disease was evaluated as AUC=0.648 for chronic ITP, with an optimal cut-off value of 4.5 days, a sensitivity of 65.4%, and a specificity of 62.5%.Lymphocyte counts was evaluated as AUC(area under the ROC curve)=0.648 for chronic ITP, the optimal cut-off value was 3.01×109/L, the sensitivity was 59.4%, and the specificity was 78.2%.The initial course of disease and lymphocyte count had no significant value in evaluation of the treatment outcome (P<0.05). The initial treatment of patients with increased megakaryocyte counts was better than those with the normal level, and the difference was statistically significant(Z=6.051, P<0.05).@*Conclusion@#The initial course of disease and lymphocyte count can help to assess the duration of ITP in children.Patients with increased bone marrow megakaryocyte counts can achieve better results at initial treatment.
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Objective To explore the predictive factors for determining the therapeutic response and prognosis of severe thrombocytopenia (TP) in patients with primary Sj(o)gren syndrome(pSS).Method Patients with pSS and severe TP (platelet count ≤ 50× 109/L) admitted between 2010 to 2016 at Peking Union Medical College Hospital were classified according to their therapeutic response and analyzed retrospectively.The response parameters and clinical data including bone marrow aspiration results and laboratory findings were collected.Result Thirty patients were finally analyzed,including twenty with appreciable bone marrow aspiration results.Fourteen and 7 patients achieved a complete response (CR) and a partial response (PR) respectively,other 9 patients with no response (NR).The megakaryocyte counts in bone marrow (BM-MK) counts per slide in each group were 13.0 (9.2,23.5) in CR patients,7.0 (7.0,20.0) in PR patients,and 5.0 (1.0,6.0) in NR patients.BM-MK counts in patients with clinical response (CR+PR)were significantly higher than those with NR (P=0.006).A receiver-operation characteristic analysis revealed a cutoff value of BM-MK counts at 6.5 per slide stratifying patients by different responses with a sensitivity of 13/14,a specificity of 6/7,and area under the curve of 0.879.Univariate analysis indicated a better prognosis as BM-MK counts＞6.5 per slide.Conclusion BM-MK count could be a predictive factor of response in patients with pSS and severe TP.Patients with BM-MK counts≤6.5 per slide represent worse platelet improvement..
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Objective@#To observe the effect of poloxamer 188 (P188) on megakaryocyte cultivation and induction from cord blood mononuclear cells in order to obtain more megakaryocyte progenitor cells (MPC).@*Methods@#The cord blood mononuclear cells were isolated and inoculated in cell culture bag or cell culture flask respectively. The WIGGENS shaker and cell culture bags were used to mimick WAVE Bioreactor for three-dimensional (3D) cell culture, and the P188 was added to induction medium, The cells were detected for morphology, surface marker, viability, and number on day 14.@*Results@#In the two-dimensional (2D) culture, CD41+, CD41+/CD61+, CD61+ megakaryocytic numbers increased significantly after adding P188 (all P<0.01). And in the 3D culture of adding P188, the cell volume became larger and the nuclear shape was irregular, the cytoplasm appeared magenta granules, and the megakaryocyte cells became more mature. By 3D culture, the expression of CD41/CD61 was (36.30±1.27)% vs (23.95±1.34)%, hence the differentiation for MPC was significantly higher than that in the 2D group (P<0.01). Furthermore, adding P188 in 3D culture resulted in highest differentiation efficiency for MPC [(59.45±1.20)%]. There were no significantly differences in terms of cell viability and cell number among 3D culture containing P188, 2D and 3D culture groups (all P>0.05).@*Conclusion@#3D culture was beneficial for the differentiation of MPC, but the cell viability was lower than 2D group; However, the satisfied cell growth and better induction efficiency were obtained by adding of P188, which might provide a new method of megakaryocytes production for clinical application.
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Objective To analyze the risk factors related to adult patients of primary immune thrombocytopenia with infection,so as to provide basis for control and prevention of infection. Methods From January 2015 to December 2017,the clinical data of 166 adult patients of primary immune thrombocytopenia in the People's Hospital of Shaoxing-were retrospectively analyzed. Statistical analysis was performed by SPSS21. 0 statistical software. Results The infec-tion rate was 24. 70％ (41 / 166). The risk factors related to infection were age(OR = 1. 290,P = 0. 011),the number of bone marrow megakaryocytes(OR = 1. 220,P = 0. 022),diabetes(OR = 2. 163,P < 0. 001) and glucocorticoid (OR =2. 203,P <0. 001). There were 45 strains of pathogenic bacteria detected in 41 patients,gram negative bacteria accounted for 51. 11％ (23 / 45),gram positive bacteria accounted for 44. 44％ (20 / 45),and fungi accounted for 4. 44％ (2 / 45). Conclusion A variety of factors are related to infection in adult primary immune thrombocytopenia patients. It is necessary to take effective prevention measure,which will help reduce the incidence of infection.
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Objective: To observe the effect of poloxamer 188 (P188) on megakaryocyte cultivation and induction from cord blood mononuclear cells in order to obtain more megakaryocyte progenitor cells (MPC). Methods: The cord blood mononuclear cells were isolated and inoculated in cell culture bag or cell culture flask respectively. The WIGGENS shaker and cell culture bags were used to mimick WAVE Bioreactor for three-dimensional (3D) cell culture, and the P188 was added to induction medium, The cells were detected for morphology, surface marker, viability, and number on day 14. Results: In the two-dimensional (2D) culture, CD41(+), CD41(+)/CD61(+), CD61(+) megakaryocytic numbers increased significantly after adding P188 (all P<0.01). And in the 3D culture of adding P188, the cell volume became larger and the nuclear shape was irregular, the cytoplasm appeared magenta granules, and the megakaryocyte cells became more mature. By 3D culture, the expression of CD41/CD61 was (36.30±1.27)% vs (23.95±1.34)%, hence the differentiation for MPC was significantly higher than that in the 2D group (P<0.01). Furthermore, adding P188 in 3D culture resulted in highest differentiation efficiency for MPC [(59.45±1.20)%]. There were no significantly differences in terms of cell viability and cell number among 3D culture containing P188, 2D and 3D culture groups (all P>0.05). Conclusion: 3D culture was beneficial for the differentiation of MPC, but the cell viability was lower than 2D group; However, the satisfied cell growth and better induction efficiency were obtained by adding of P188, which might provide a new method of megakaryocytes production for clinical application.
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Bioreactors , Cell Differentiation , Cells, Cultured , Fetal Blood , Megakaryocytes , PoloxamerABSTRACT
Objective To investigate the effect of microRNA-223-3p (miR-223-3p) on megakaryocytic differentiation and maturation, and explore the potential mechanism .Methods The endogenous expression of miR-223-3p during megakaryocyte ( MK) differentiation was detected by real-time PCR.Flow cytometry further indicated that alteration of miR-223-3p in human cell lines exerted effects on MK differentiation and maturation .By performing integrative bioinformatic analysis, the potential miR-223-3p target gene, MYH10,was identified.Real-time PCR, luciferase reporter assay and flow cytometry revealed that MYH10 was a direct target of miR-223-3p.Results Endogenous expression of miR-223-3p was in-creased with the differentiation and maturation of MK .The expression of megakaryocytic surface markers CD41 and CD61 and the ploidy were significantly increased in K562 and Meg-01 cells after transfection with miR-223-3p mimics.The expression of MYH10 decreased with the increase in miR-223-3p.Using a luciferase reporter assay ,we demonstrated that MYH10 was a direct target of MiR-223-3p.Furthermore, direct downregulation of MYH10 promoted MK polyploidization . Conclusion MiR-223-3p might regulate the polyploidization of MK by targeting MYH10.
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Objective To investigate the effect of microRNA-223-3p (miR-223-3p) on megakaryocytic differentiation and maturation, and explore the potential mechanism .Methods The endogenous expression of miR-223-3p during megakaryocyte ( MK) differentiation was detected by real-time PCR.Flow cytometry further indicated that alteration of miR-223-3p in human cell lines exerted effects on MK differentiation and maturation .By performing integrative bioinformatic analysis, the potential miR-223-3p target gene, MYH10,was identified.Real-time PCR, luciferase reporter assay and flow cytometry revealed that MYH10 was a direct target of miR-223-3p.Results Endogenous expression of miR-223-3p was in-creased with the differentiation and maturation of MK .The expression of megakaryocytic surface markers CD41 and CD61 and the ploidy were significantly increased in K562 and Meg-01 cells after transfection with miR-223-3p mimics.The expression of MYH10 decreased with the increase in miR-223-3p.Using a luciferase reporter assay ,we demonstrated that MYH10 was a direct target of MiR-223-3p.Furthermore, direct downregulation of MYH10 promoted MK polyploidization . Conclusion MiR-223-3p might regulate the polyploidization of MK by targeting MYH10.
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Objective To observe the curative effect in patients with systemic lupus erythematosus(SLE) complicated by moderate or severe thrombocytopenia with a blood platelet count(BPC) of under 50 × 109/L and analyze its related factors. Methods We retrospectively analyzed the clinical data on 109 SLE patients with mod-erate or severe thrombocytopenia. Results Of the 109 patients,82(75.2%)had complete response(CR),15 (13.8%)had partial response(PR),and 12(11.0%)had no response(NR),respectively. As compared with the CR+PR group,the NR group had a higher incidence rate of decreased bone marrow megakaryocyte(P 0.05 for all comparisons). The total effectiveness rate did not differ signifi-cantly between MP pulse therapy and high-dose corticosteroid therapy. Conclusions A decrease in bone marrow megakaryocytes can be an adverse factorfor affecting the efficacy in patients with SLE complicated by moderate or severe thrombocytopenia.
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Tal vez por haber sido consideradas como simples restos citoplasmáticos de los megacariocitos encargadas únicamente de la reparación de heridas, las plaquetas han tenido un lugar secundario en cuanto a su estudio e interés en comparación con los otros componentes celulares de la sangre. Sin embargo, en los últimos 20 años se ha avanzado mucho en el conocimiento de estas fascinantes células que de a poco han recobrado un lugar destacado dentro de la hematología. A lo largo de este trabajo se han revisado los aportes más destacados y novedosos acerca del proceso de biogénesis plaquetaria, su regulación por el microambiente medular y factores humorales, recorriendo desde la generación de megacariocitos hasta la liberación de plaquetas libres.
Perhaps for being considered mere megakaryocyte cytoplasmic debris responsible for wound repair alone, platelets have had a secondary role when compared to other cellular blood components. However, in the last 20 years we have learned much more about these fascinating cells, which have slowly regained a prominent place in hematology. This review discusses the most outstanding and novel contributions on platelet biogenesis, its regulation by the bone marrow microenvironment and humoral factors, analyzing from megakaryocyte generation to platelet release.
Talvez por ter sido considerados simples restos citoplasmáticos dos megacariócitos, encarregadas apenas da reparação de feridas, as plaquetas têm tido um lugar secundário quanto a seu estudo e interesse em comparação com os outros componentes celulares do sangue. Entretanto, nos últimos 20 anos foi possível aprender muito a respeito destas fascinantes células que aos poucos foram recobrando um lugar de destaque dentro da hematologia. Ao longo deste trabalho foram revistas as contribuições mais destacadas e novas acerca do processo de biogênese plaquetária, sua regulação pelo microambiente medular e fatores humorais, percorrendo desde a geração de megacariócitos até a liberação de plaquetas livres.
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Female , Megakaryocytes , Cells , Origin of Life , Cytoplasm , HematologyABSTRACT
Objective To evaluate the value and significance of testing megakaryocyte micronuclei in bone marrow smears for hematopathy diagnosis.Methods Bone marrow smears from a total of 863 cases of patients with hemopathy were collected from 2002 to 2009 at the second affiliated hospital of zhejiang university school of medicine.Smears from 25 healthy individuals were used as control.The bone marrow smears were subjected to Wright-Giemsa staining.The number of megakaryocytes, morphous and positive rate of megakaryocyte micronucleis were recorded by using low power lens and immersion objective.Statistical differences of positive rate in the different diseases, pathologic subtypes, total number and type of megakaryocyte were analyzed.Results The megakaryocyte micronucleis were round or oval with varying size, distributed in paranuclear or away from nuclear and even free in extracellular, which could be observed in large and medium megakaryocytes.The positive rate of megakaryocyte micronucleis was highest in myeloid leukemia, particularly in the subtype of M6, M2, M4 and M5b with dyshaematopoiesis.Megakaryocyte micronucleis could also be found in MA, infection and benzolism, but rarely observed in the lymphocytic leukemia.Conclution The detection of megakaryocyte micronuclei was related with its amount, size and type.There is significant difference of the postive rates of megakaryocyte micronuclei testing among the different hematopathies and pathologic subtypes.Megakaryocyte micronuclei testing should be valuable in the the diagnoses of hematopathy.
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Thrombocytopenia is a common clinical problem with many etiological causes. Although transient bone marrow suppression and marrow infiltration by malignancies are important causes, certain non-malignant conditions such as nutritional causes and infections are equally important as the treatment is simple and cure is possible. Depending on the aetiology, the clinical presentation may vary. Knowing the exact aetiology is important for specific treatment and prognostication. A total of 303 cases of thrombocytopenia were studied out of which males were 44% and females were 56%. The patients’ age ranged from 5 months to 84 yrs. The commonest presenting symptom was fever with bleeding manifestations and jaundice. 14% of cases are of Grade 1, 20% of the cases of Grade 2, 5% of cases of Grade 3 and 31% of cases had counts less than 25000/cu.mm i.e. of Grade 4. 50% of cases in Grade 4 had a decreased number of megakaryocytes in the bone marrow. The most common cause of thrombocytopenia in our study was megaloblastic anaemia (48.6%), ITP (20%), post-viral (10.9%) followed by leukaemia, aplastic anaemia and others. Thrombocytopenia has a spectrum of causes which can be diagnosed by detailed history and peripheral smear examination supported by bone marrow examination. Megaloblastic anaemia was the commonest cause of thrombocytopenia followed by immune thrombocytopenia. Unlike in the western India megaloblastic anemia is highly prevalent and is the leading correctable cause of thrombocytopenia. Most of the patients with Grade 4 thrombocytopenia had a decreased number of megakaryocytes in the bone marrow suggesting a production defect.
ABSTRACT
Megakaryocytic hematopoietic disorders is one of the leading causes of death in patients with acute radiation sickness bleeding. Thrombopoietin (TPO),the main stimulation factor of megakaryocytopoiesis, can promote megakaryocytic hematopoietic recovery after radiation injury and increase peripheral platelet count. Early application of TPO after irradiation can play a key role in prevention and treatment of bleeding complications of acute radiation sickness. Studies have shown that TPO may have a stronger role in promoting hematopoiesis. In this paper, a brief overview of new progress on the TPO and acute radiation sickness is summarized.
ABSTRACT
Megakaryocytic hematopoietic disorders is one of the leading causes of death in patients with acute radiation sickness bleeding. Thrombopoietin (TP0)the main stimulation factor of megakaryocytopoiesis ’ can promote megakaryocytic hematopoietic recovery after radiation injury and increase peripheral platelet count. Early application of TP0 after irradiation can play a key role in prevention and treatment of bleeding complications of acute radiation sickness. Studies have shown that TP0 may have a stronger role in promoting hematopoiesis. In this paper,a brief overview of new progress on the TP0 and acute radiation sickness is summarized.