MDS Group Research Articles

Expression and Significance of PD-1/PD-L1 in MDS Blast Cells, T Lymphocyte Cell Subsets and Treg Cells

To investigate the expression and significance of PD-1/PD-L1 in MDS blast cells, T lymphocyte cell subsets and Treg cells. Eighty-eight MDS patients and 19 AML patients were collectd as the study subjects, and Iron deficiency anemia and healthy bone marrow donors were used as control group. The expression of PD-1/PD-L1 in MDS/AML blast cells, T lymphocyte cell subsets and Treg cells was detected by flow cytometry, and the expression level of Th1/Th2/Th17-related cytokines in peripheral serum was detected. The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells in low risk MDS group were lower than that in control group, medium and high risk MDS group and AML group（all P < 0.01）, and Th1/Th17 type cytokines were dominant. The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells of intermediate and high risk MDS group and AML group were higher than that of control group and low risk MDS group (all P < 0.01), and Th2 type and Treg type (IL-10、TGF-β) cytokines were dominant. After treatment, the differences of PD-1/PD-L1 expression were not statisticatly significant in blast cells, T lymphocyte cell subsets and Treg cells between the MDS remission group and the control group (all P >0.05). The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells in MDS non-remission group were significantly higher than that in remission group and control group (all P < 0.01). The high expression of PD-1/PD-L1, dominance of Treg (IL-10、TGF-β) and Th2-related cytokines and inhibition of effector T lymphocyte cells in patients with MDS is conducive to tumor cell proliferation and immune escape, which may promote the progression of MDS disease.

Complete Blood Counts and Research Parameters in the Detection of Myelodysplastic Syndromes.

100 samples from healthy individuals were used to establish the values of research parameters in normal subjects. A retrospective study was conducted including 66 patients diagnosed with MDS, 90 cytopenic patients due to other diseases (cancer patients receiving therapy, aplastic anemia, other hematological malignancies) and 50 with macrocytic anemia. The Wilcoxon test was applied to detect statistical differences among the groups of patients, considering p < 0.05 significant. The diagnostic performance of the RUO parameters for discriminating MDS among cytopenias was evaluated using receiver operating characteristic (ROC) curve analysis. Amultivariable logistic regression model was performed to identify the potential predictors for having MDS. The area under curve (AUC) and the Hosmer-Lemeshow test of the model were assessed. The performance of the model was verified in a prospective study including 224 cytopenic patients (validation group). In the MDS group, the mean cell volume (MCV), percentage of macrocytic red cells (MAC), red cell distribution width (RDW) and immature platelets fraction (IPF) had increased values compared to the cytopenic and normal patients, while platelets, red and white cell counts, Neu X (related to the cytoplasmic complexity of neutrophils), Neu Y (related to nucleic acid content) and Neu Z (related to cell size) were lower (p < 0.001). Neu X, Neu Y, and Neu Z showed higher AUC for detecting MDS > 0.80; MAC, RDW and IPF AUC > 0.76. The multivariable model demonstrated that Neu X and Neu Y could be used in the recognition of MDS, AUC 0.88. In the validation group, 89.0% of the MDS patients were well classified. MDS are common malignant disorders with a poor prognosis, and early diagnosis is warranted for optimal benefit from treatment. RUO gain insights to detect dysplasia of MDS and could be used in the differential diagnosis of MDS from cytopenias of other etiologies.

Real-world evidence from a retrospective study on suicide during depression: clinical characteristics, treatment patterns and disease burden

BackgroundSuicide stands as both a primary symptom and the direst outcome of major depressive disorder (MDD). The scarcity of effective treatment strategies makes managing MDD patients with suicide especially challenging. Hence, it is crucial to investigate disease characteristics and efficacious therapeutic strategies for these patients, drawing insights from disease databases and real-world data.MethodsIn this retrospective study, MDD patients hospitalized between January 2013 and December 2020 were investigated using Electronic Health Records (EHR) data from Beijing Anding Hospital. The study enrolled 4138 MDD patients with suicidal ideation or behavior (MDS) and 3848 without (MDNS). Demographic data, clinical attributes, treatment approaches, disease burden, and re-hospitalization within one year of discharge were extracted and compared.ResultsPatients in the MDS group were predominantly younger and female, exhibiting a higher prevalence of alcohol consumption, experiencing frequent life stress events, and having an earlier onset age. Re-hospitalizations within six months post-discharge in the MDS group were significantly higher than in the MDNS group (11.36% vs. 8.91%, p < 0.001). Moreover, a more considerable fraction of MDS patients underwent combined electroconvulsive therapy treatment (56.72% vs. 43.71%, p < 0.001). Approximately 38% of patients in both groups were prescribed two or more therapeutic regimes, and over 90% used antidepressants, either alone or combined. Selective serotonin reuptake inhibitors (SSRIs) were the predominant choice in both groups. Furthermore, antidepressants were often prescribed with antipsychotics or mood stabilizers. When medication alterations were necessary, the favoured options involved combination with antipsychotics or transitioning to alternative antidepressants. Yet, in the MDS group, following these initial modifications, the addition of mood stabilizers tended to be the more prioritized alternative.ConclusionsMDD patients with suicidal ideation or behaviour displayed distinctive demographic and clinical features. They exhibited intricate treatment patterns, a pronounced burden of illness, and an increased likelihood of relapse.

Cardiovascular risk in elderly Egyptians with myelodysplastic syndromes

BackgroundMyelodysplastic syndromes (MDS) are a heterogeneous group of clonal stem cell-derived disorders manifested by cytopenias peripherally. MDS initiates an inflammatory process which lead to atherosclerosis. Patients with MDS have 1.5-fold higher risks of cardiovascular risks.ObjectiveEvaluation of cardiovascular risk and unraveling the possible correlation of carotid intima media thickness (CIMT) with clinical, hematologic, and laboratory variables in elderly patients with MDS.MethodsWe conducted a case–control research on 168 participants to elicit the serum level of homocysteine, serum high-sensitive C-reactive protein (Hs-CRP), and CIMT in relation to cardiovascular risk, 84 of whom had MDS diagnoses. From March 2021 to the end of May 2023, we chose them among the inpatients and outpatients of the Menoufia University Hospitals’ Clinical Hematology Division. We contrasted the chosen MDS patients with 84 healthy individuals as controls.ResultsRight and left CIMT were significantly higher in MDS patients than controls as well as for lipid profile, blood pressure, blood sugar, and Hs-CRP. Homocysteine was higher in MDS patients but of no statistical significance.ConclusionAs CIMT and C-reactive protein level were significantly higher in studied patients, we may consider MDS group of patients to have higher risk for cardiovascular disease than normal people.

Effect of Conditioning Regimens and Graft-versus-Host Disease Prophylaxis on the Outcomes of Umbilical Cord Blood Transplantation Performed with Cyclophosphamide/Total Body Irradiation-Based Regimens

Umbilical cord blood (UCB) is a valuable alternative donor source for allogeneic hematopoietic stem cell transplantation. Various conditioning regimens and graft-versus-host disease (GVHD) prophylaxis regimens aimed at improving the outcomes of umbilical cord blood transplantation (UCBT) have been explored; however, the differences in their effects remain unclear. This study was conducted to elucidate the differences in the effects of conditioning and GVHD prophylaxis regimens on UCBT outcomes by disease type in a nationwide, retrospective study. We retrospectively analyzed the effects of conditioning and GVHD prophylaxis regimens on the outcomes of UCBT performed with cyclophosphamide (Cy)/total body irradiation (TBI)-based regimens in patients with acute myeloid leukemia (AML; n=1126), acute lymphoblastic leukemia (ALL; n=620), myelodysplastic syndrome (MDS; n=170), and lymphoma (n=128). Multivariate analysis for overall survival (OS) demonstrated the benefit of adding high-dose cytarabine to the Cy/TBI regimen for the AML group (relative risk [RR], .76; P=.003) and lymphoma group (RR, .54; P=.02), but not for the ALL and MDS groups. In the ALL group, adding etoposide to the Cy/TBI regimen was associated with a lower OS (RR, 1.45; P=.03). For GVHD prophylaxis, a tacrolimus/methotrexate regimen was associated with a lower OS compared with a cyclosporine/methotrexate regimen in the AML group (RR, 1.26; P=.01); this difference was not observed in the other groups. These differences in OS according to the conditioning and GVHD prophylaxis regimen were attributable mainly to differences in relapse risk. Our data show that the effects of conditioning regimens and GVHD prophylaxis on UCBT outcomes differed according to disease type. UCBT outcomes could be improved by selecting optimal conditioning regimens and GVHD prophylaxis for each disease type.

Next-Generation Sequencing Testing and Triaging of Bone Marrow Samples: Indications, Pre-authorization and Medical Necessity for Patient Care

Abstract Introduction/Objective Bone marrow biopsy (BMBx) is an invasive procedure and samples are usually obtained for diagnosis, staging and prognosis of hematolymphoid disorders (HDs). Given the need for WHO and ICC classification of HDs; the Next Generation Sequencing (NGS) testing (in addition to other esoteric testing) has become relevant and medically necessary. It is important to triage BMBx in real-time based on clinical indications; however, pathologists are challenged by limited published guidelines as well as need for pre-authorization (PA) thus resulting in claim denials. Methods/Case Report Retrospective analysis was performed (2018 to 2022) on 1610 BMBx submitted for various clinical indications. These were categorized as follows: Cytopenias 220 (MDS); Cytosis 250 (MPN); Lymphomas work-up 350 (LY), acute leukemias 440 (AL) and plasma cell disorders 350 (PCD). In addition to esoteric testing; the decision was made to perform NGS testing based on preview of aspirate slides, clinical history and CBC. Results (if a Case Study enter NA) Providers initially ordered NGS in 10% of cases; however, NGS was added later by pathologists in 30% of BMBx samples. Abnormal NGS results were obtained as follows: MDS (70%); MPN (75%); LY (15%); AL (90%) and PCD (40%). NGS testing provided clinically useful data for MDS, MPN and AL groups. Even though LY and PCD groups also yielded positive results; the concurrent esoteric testing (Flow, FISH and Cytogenetics) was useful thus making indication of NGS in these two subset groups less indicated. Conclusion Appropriate triaging of BMBx for NGS testing is medically necessary for following reasons: 1) Enables precise diagnosis in a timely manner; 2) to avoid repeat BMBx if triaging guidelines are followed; and 3) decision to order NGS is made in real-time as BMBx are time sensitive samples thus eliminating the need for PA as this study emphasize that NGS has become standard of care in the work-up of HDs.

Lessons of Nature in Trisomy-8 MDS and AML: Determinants of Clonal Drive

The clinical consequences of trisomy 8 (+8) are reflected by its categorization as an intermediate risk cytogenetic abnormality in AML while in MDS it has a more favorable impact. Gain of chromosome (chr.) 8 is a frequent event in other hematologic disorders and its prognostic role seems to be dependent on disease context. Some molecular lesions ( RUNX1, ASXL1) are distinctive of +8 AML and important in delineating its clinical phenotype and prognosis. However, the intrinsic molecular drivers contributing to clonal pathogenesis of myeloid neoplasia (MN) as repercussion of gain of chr.8 remain unclear. We stipulated that the study of clinical, genomic, and transcriptomic features of +8 MN will provide important clues to the pathogenesis of +8 and help refine its clinical implications in a specific clinical context. NGS and clinical data from a cohort of 3971 MDS and 3973 AML patients were reviewed, including normal karyotype MDS/AML (NK MDS or AML), isolated +8 (iso +8 MDS/AML), and non-isolated +8 (non-iso +8 MDS/AML). In addition, RNASeq data were available in a subcohort of patients (82: +8; 617 NK MDS). Expression of chr.8 genes was studied to unmask culprit genes in +8 MN. Compared to NK-AML, iso +8 AML patients were older (76% aged &amp;gt;60 years, P= .0005) and had more dysplastic than hyperproliferative phenotype as they presented with lower WBC (median: 9.5 vs 15.2x10 9/L, P= .008) and platelet counts (median: 49 vs 64 x10 9/L, P= .0002). Iso +8 AML had higher percentages of secondary AML (sAML, 17 vs 11%, P= .001) arising from antecedent MDS (14 vs 9%, P= .002) vs NK AML. As expected, survival analysis showed iso +8-AML to have worse median overall survival (mOS) vs NK AML (20 vs 32 months, P= .0001). Yet, when sub-classifying AML based on disease ontogeny (primary vs secondary), a significant mOS difference was only seen in primary AML (21.5 vs 34.5 months, P= .0002; in sAML, 14.7 vs 24.3 months, P= .3). As for molecular associations, a variety of mutations were more frequent in iso +8 AML vs NK AML, including: lineage transcriptional factors ( RUNX1, 33 vs 17%, P&amp;lt; .0001), chromatin modifiers ( ASXL1 37 vs 13%, P&amp;lt; .0001; EZH2 10 vs 3.4%, P&amp;lt; .0001), splicing factors ( SRSF2 30 vs 13%, P&amp;lt; .0001; U2AF1, 12 vs 3%, P&amp;lt; .0001) and cohesin genes ( STAG2, 13 vs 7%, P= .02). In contrast, mutations in FLT3 (33 vs 42%, = .01), DNMT3A (24 vs 35%, P=.002), NPM1 (14 vs 44%, P&amp;lt;.0001) and CEBPA (7 vs 12%, P= .02) were more frequent in NK AML. We then focused on the iso +8 MDS vs NK MDS groups, and interestingly, found that ASXL1 (52 vs 19%, P&amp;lt; .0001), EZH2 (25 vs 4, P&amp;lt; .0001), RUNX1 (29 vs 12%, P= .0009), and STAG2 (27 vs 6%, P&amp;lt; .0001) were more frequently found in the +8 MDS compared to NK-MDS (similar patten to AML), whereas SRSF2 (19 vs 18%, P= .9) and U2AF1 (9 vs 6%, P=.7) were similar in both. The comparison of AML and MDS then showed that iso +8 AML was more commonly associated with mutations in FLT3 (33 vs 3%, P&amp;lt; .0001), IDH1 (17 vs 5%, P= .03), and IDH2 (25 vs 3%, P&amp;lt; .0001), while iso +8-MDS had higher odds of SF3B1 (19 vs 4%, P= .001) ( Fig1A). We then investigated whether specific transcriptomic features may play a role in +8 MN pathogenesis by targeting the transcriptome of 412 chr.8 genes ( Fig1B). After adjusting for zygosity, we identified 186 genes correlating the change in expression levels with clonality. We further interrogated genes possibly eliciting oncogenic potential with log2 fold change (log2FC) ratio of at least 1.25 to reflect an extra chr.8 copy. We found higher mRNA expression levels of 13 putative culprit genes of known oncogenic potential in iso +8 MDS vs NK MDS including the anti-apoptotic protein BAG4 (LogFC 7.1 vs 5.4, P&amp;lt; .0001), possibly conferring growth advantage, toll-like receptor LY96 (LogFC 3.3 vs 2.0, P&amp;lt; .0001),the tyrosine protein kinase LYN (LogFC 8.4 vs 6.7, P&amp;lt; .0001), human oncogene WHSC1L1/NSD3 (LogFC 8.1 vs 6.5, P&amp;lt; .0001)and long non-coding RNA, PVT1 derived from an 8q24 intergenic region (LogFC 4.0 vs 2.2, P&amp;lt; .0001). In summary, iso +8 MN represents a distinct entity defined by older age and dysplastic phenotype, likely due to genotypic enrichment in RUNX1 mutations, while inversely associated with proliferation genes ( e.g., FLT3 and NPM1 in NK). The key culprit oncogenes implicated in +8 MN may be located on the extra chr.8 itself. Indeed, we found transcriptomic changes pertinent to +8 MDS whose role in driving leukemogenesis and interaction with specific myeloid drivers needs to be confirmed by future experimentations on cellular and murine models.

Accurate stratification between VEXAS syndrome and differential diagnoses by deep learning analysis of peripheral blood smears.

VEXAS syndrome is a newly described autoinflammatory disease associated with UBA1 somatic mutations and vacuolization of myeloid precursors. This disease possesses an increasingly broad spectrum, leading to an increase in the number of suspected cases. Its diagnosis via bone-marrow aspiration and UBA1-gene sequencing is time-consuming and expensive. This study aimed at analyzing peripheral leukocytes using deep learning approaches to predict VEXAS syndrome in comparison to differential diagnoses. We compared leukocyte images from blood smears of three groups: participants with VEXAS syndrome (identified UBA1 mutation) (VEXAS); participants with features strongly suggestive of VEXAS syndrome but without UBA1 mutation (UBA1-WT); participants with a myelodysplastic syndrome and without clinical suspicion of VEXAS syndrome (MDS). To compare images of circulating leukocytes, we applied a two-step procedure. First, we used self-supervised contrastive learning to train convolutional neural networks to translate leukocyte images into lower-dimensional encodings. Then, we employed support vector machine to predict patients' condition based on those leukocyte encodings. The VEXAS, UBA1-WT, and MDS groups included 3, 3, and 6 patients respectively. Analysis of 33,757 images of neutrophils and monocytes enabled us to distinguish VEXAS patients from both UBA1-WT and MDS patients, with mean ROC-AUCs ranging from 0.87 to 0.95. Image analysis of blood smears via deep learning accurately distinguished neutrophils and monocytes drawn from patients with VEXAS syndrome from those of patients with similar clinical and/or biological features but without UBA1 mutation. Our findings offer a promising pathway to better screening for this disease.

Myeloid-derived suppressor cells inhibit natural killer cells in myelodysplastic syndromes through the TIGIT/CD155 pathway

ABSTRACTObjectiveThis experiment will explore the role of TIGIT/PVR signaling pathway in the pathogenesis of MDS immune tolerance through in vitro co-culture of NK cells and MDSC cells.MethodsFlow cytometry was used to detect the expression percentage of MDSCs and CD155 on MDSCs in the bone marrow of MDS patients and controls. The expression of NK cell surface receptors (NKG2D, NKp30, NKp46), secreted cytokines (perforin, granzyme B, CD107a, IFN-γ) and NK cell apoptosis rate were detected by flow cytometry to evaluate the effect of MDSCs on NK cell function.ResultsThe number of MDSCs in bone marrow of MDS patients was notably higher than that of the control group (8.39 ± 7.01 vs 2.31 ± 1.65, P = 0.0001). Compared with the control group, the expression of CD155 on MDSCs in MDS group was increased (31.81 ± 21.33 vs. 10.49 ± 6.53, P < 0.0001). After NK cells were co-cultured with MDSCs, NKG2D, NKp30, NKp46, CD107a, IFN-γ, perforin and granzyme B were decreased, and the NK function partially recovered after the addition of inhibitors.ConclusionCompared with the normal control, MDSCs and CD155 on MDSCs were highly expressed in MDS patients. After co-culture with MDSCs, the expression of NK cells’ surface receptors decreased, the secretion of cytokines decreased and the apoptosis rate increased. After blocking TIGIT/CD155 pathway, NK cell function was reversed, but NK cell apoptosis was not reduced.

Venetoclax and Azacytidine Therapy in High-Risk Myelodysplastic Syndromes: A Retrospective Evaluation of a Real-World Experience. Latin-American MDS Group - Glam

Background: Venetoclax, a selective, potent, orally bioavailable BCL2 inhibitor, has shown synergistic activity with hypomethylating agents in preclinical and clinical studies in high-risk MDS and AML. The primary objective of this study was to describe the safety, effectiveness, and management of this therapy in a real-world scenario. Methods: High-risk MDS patients receiving 400 mg of oral venetoclax daily for the first 14 days of each 28-day cycle, combined with azacytidine (AZA) at 75 mg/m2 from days 1 to 7 of the 28-day cycle were reported, and retrospectively analyzed. The response rates were evaluated according to IWG-2016. A Log-Rank test (Mantel Cox) and Kaplan-Meier were performed to compare the OS of responders and non-responders. Results: From September 2017 to July 2022, 18 high-risk MDS patients under venetoclax and azacytidine were retrospectively reported. This Latin-American multicenter study showed a median age of 63 (R 37-81); 61% male; 16% median blasts at diagnosis (R 0-19); an MDS subtype of MDS_EB (72%); therapy-related MDS (11%); IPSS-R > 4.5 (83%); and high, and very high-risk cytogenetic subgroups (53%) at diagnosis. Fifteen patients (83%) were naïve for hypomethylating agents. With a median cycle of 4.5 (R 1-39), the overall response rate was 78% (Complete Remission 33%, Partial Remission 17%, medular Complete Remission 17%, and Hematology Improvement 11%) and 22% for non-responders (Stable Disease and Progressive Disease). A total of 7 (39%) patients underwent transplantation, and the median number of cycles to reach the transplant was 1 (R 1-5). The estimated median overall survival (months) for responders versus non-responders was 39.1 vs 4.5 respectively (p=0.024). Five patients (28%) progressed to AML, and there was no correlation with the type and depth of the response achieved (Graphic). The most common treatment-emergent adverse events were neutropenia and infectious diseases (22%). Dose reductions or adjustments to azoles were necessary in 10 patients (56%). There were 7 deaths, 6 of which were related to MDS: infections (3), hemorrhage (1), and AML (2). One patient in Complete Remission died because of Covid-19. Conclusion: Combination therapy of venetoclax and azacytidine in high-risk MDS patients demonstrated a tolerable safety profile, a frequent need of dose adjustments, a high overall response rate (78%), a statistically significant longer overall survival of responders (p=0.024) and a high efficacy as bridge-therapy to transplant. Further experiences are warranted. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Therapy Related Myeloid Neoplasms Following PARP Inhibitors : Real-Life Experience

Purpose: To assess the hematologic toxicity of PARP inhibitors (PARPi) treatment, and provide insights into the diagnosis and management of therapy-related myeloid neoplasms (t-MN) following PARPi. Experimental Design: In a leading cancer center, we compared the profile of 13 t-MN diagnosis among 37 ovarian cancer (OC) patients treated with PARPi referred for hematological consultation. Next, we compared these 13 t-MN post PARPi among 37 t-MN post OC. Finally, we described the specificities of 69 t-MN post PARPi treated for gynecologic cancer (breast and/or ovarian cancer) in a national observatory. Results: From 2016 to 2021, cumulative incidence of t-MNs was 3.5% (13/373) among OC patients treated with a PARPi. At the time of referral for hematological consultation, patients who developed t-MNs had a longer PARPi exposure (9 months vs. 3, p= 0.01), lower platelet count (74 vs. 173 G/L, p=0.0005), and more cytopenias (2 vs 1, p=0.0005). After NGS evaluation of patients without t-MN, 5 patients (13.5%) were finally classified as CHIP (clonal hematopoiesis of indeterminate potential), 7 (18.9%) as CCUS (clonal cytopenia of undetermined significance) and 2 (5%) as ICUS (idiopathic cytopenia of undetermined significance). Compared to t-MN not exposed to PARPi, t-MN-PARPi patients had more BRCA1/2 germline mutation (61.5% vs 0% p=0.03) but similar OS [8.2 (95% CI: 2.03-18.7) months in the t-MN-PARPi group compared to 6.1 (95% CI: 3-12.2) months in the t-MN-non-PARPi group; p=0.8]. In the national observatory, t-MN post PARPi had mostly a complex karyotype (61%) associated with a high rate of TP53 mutation (71%) with a median VAF of 41% (IQR, 17-67) and 75% of multi-hit mutations (Figure 1). IPSS score was "Low and Intermediate-1" and "Intermediate-2 and High" MDS groups for 50% of patients, respectively. AMLs were mainly considered as adverse (75%) according to the ELN 2017 classification. Median OS from t-MN diagnosis was 9.7 months (95% CI: 5.3-13.9). In multivariate analysis, a longer time between the end of PARPi and t-NM (HR 1.046, p=0.02), olaparib treatment compared to others PARPi (HR 5.82, p=0.003) and AML (HR 2.485, p=0.01) were associated with shorter OS independently of the BRCA1/2 status (Figure 2). Conclusions: In a large series, we described a high incidence of t-MN post PARPi associated with unfavorable cytogenetic and molecular abnormalities leading to poor OS. Time to diagnose t-MN after PARPi interruption seems to influence OS, suggesting the importance of early diagnosis. Particular consideration should be done on BRCA1/2 patients, and in case of thrombocytopenia or more than one cytopenia and relatively long PARPi exposure. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Influencing factors of iron metabolism assessment in patients with myelodysplastic syndrome: A retrospective study

目的分析骨髓增生异常综合征（MDS）患者铁代谢评估的影响因素。方法181例MDS患者接受磁共振成像（MRI）和（或）能谱CT（DECT）对肝脏和心脏铁浓度的检测，其中41例患者在2次检查期间接受了规律铁螯合治疗（ICT）。同步检测调整铁蛋白（ASF）、红细胞生成素（EPO）、心功能、肝转氨酶、肝炎抗体、外周血T细胞极化等指标，并收集患者是否合并骨髓纤维化、脾大、环孢素A使用等信息进行比较分析。结果MRI组、DECT组肝铁浓度均与ASF呈正相关（r分别为0.512、0.606，P值均<0.001），MRI组心铁浓度与ASF仅呈弱相关（r＝0.303，P<0.001），而DECT组心铁浓度与ASF无明显相关性（r＝0.231，P＝0.053）。输血依赖显著影响患者肝和心铁浓度［MRI组：LIC：（28.370±10.706）mg/g对（7.593±3.508）mg/g，t＝24.30，P<0.001；MIC：1.81对0.95，z＝2.625，P<0.05，DECT组：肝VIC：（4.269±1.258）g/L对（1.078±0.383）g/L，t＝23.14，P<0.001：心VIC：1.69对0.68，z＝3.142，P<0.05］。重度以上铁过载组患者EPO浓度明显高于轻中度铁过载组及正常组（P值均<0.001）；MDS伴环状铁粒幼红细胞（MDS-RS）患者与和其他MDS低危组患者相比，肝铁浓度明显增高［DECT组：3.80（1.97,5.51）g/L对1.66（0.67,2.94）g/L，P＝0.004；MRI组：13.7（8.1,29.1）mg/g对11.6（7.1,21.1）mg/g，P＝0.032］。而年龄、骨髓纤维化、脾大、T细胞极化、环孢素A的使用、肝转氨酶、肝炎抗体阳性等因素对铁代谢无明显影响。结论MDS患者肝铁浓度与ASF呈正相关，心铁浓度与ASF无明显相关性。输血依赖、EPO浓度、合并RS是铁代谢的影响因素。

Dysfunctional bone marrow endothelial progenitor cells are involved in patients with myelodysplastic syndromes

BackgroundMyelodysplastic syndromes (MDS) are a group of heterogeneous myeloid clonal disorders characterized by ineffective haematopoiesis and immune deregulation. Emerging evidence has shown the effect of bone marrow (BM) endothelial progenitor cells (EPCs) in regulating haematopoiesis and immune balance. However, the number and functions of BM EPCs in patients with different stages of MDS remain largely unknown.MethodsPatients with MDS (N = 30), de novo acute myeloid leukaemia (AML) (N = 15), and healthy donors (HDs) (N = 15) were enrolled. MDS patients were divided into lower-risk MDS (N = 15) and higher-risk MDS (N = 15) groups according to the dichotomization of the Revised International Prognostic Scoring System. Flow cytometry was performed to analyse the number of BM EPCs. Tube formation and migration assays were performed to evaluate the functions of BM EPCs. In order to assess the gene expression profiles of BM EPCs, RNA sequencing (RNA-seq) were performed. BM EPC supporting abilities of haematopoietic stem cells (HSCs), leukaemia cells and T cells were assessed by in vitro coculture experiments.ResultsIncreased but dysfunctional BM EPCs were found in MDS patients compared with HDs, especially in patients with higher-risk MDS. RNA-seq indicated the progressive change and differences of haematopoiesis- and immune-related pathways and genes in MDS BM EPCs. In vitro coculture experiments verified that BM EPCs from HDs, lower-risk MDS, and higher-risk MDS to AML exhibited a progressively decreased ability to support HSCs, manifested as elevated apoptosis rates and intracellular reactive oxygen species (ROS) levels and decreased colony-forming unit plating efficiencies of HSCs. Moreover, BM EPCs from higher-risk MDS patients demonstrated an increased ability to support leukaemia cells, characterized by increased proliferation, leukaemia colony-forming unit plating efficiencies, decreased apoptosis rates and apoptosis-related genes. Furthermore, BM EPCs induced T cell differentiation towards more immune-tolerant cells in higher-risk MDS patients in vitro. In addition, the levels of intracellular ROS and the apoptosis ratios were increased in BM EPCs from MDS patients, especially in higher-risk MDS patients, which may be therapeutic candidates for MDS patients.ConclusionOur results suggest that dysfunctional BM EPCs are involved in MDS patients, which indicates that improving haematopoiesis supporting ability and immuneregulation ability of BM EPCs may represent a promising therapeutic approach for MDS patients.

Effect of 12 Weeks Modified “Dantien-Salee” Yoga Training on Pulmonary Function, Functional Fitness, and Quality of Life in Elderly Thai

Objective: To investigate the effect of 12 weeks Modified “Dantien-Salee” Yoga training (MDS) on pulmonary function, functional fitness, and quality of life (QoL) in elderly Thai. Material and Methods: The quasi-experimental study was conducted using a sample of twenty-four elderly subjects with no previous experience of yoga training. They were equally divided into a MDS group and control group. The MDS group were asked to attend a 70-minite session of MDS training three times a week over 12 weeks. The control group was asked to continue their normal daily routine over the same period. Pulmonary function tests, functional fitness assessments, and QoL by SF-36 assessments, were all measured at baseline and after the 12 weeks study period. A two-way repeated measures ANOVA was used to evaluate differences between groups. Results: There were no differences in spirometry and lung volumes over the intervention period. Both maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP) showed tendency to improve in the MDS group but did not reach statistically significant level. Functional fitness and QoL improved significantly (p&lt;0.05) over the 12 weeks period in MDS group. Conclusion: MDS program led to improvement of functional fitness and enhance the QoL in the study group of elderly participants without significant change in pulmonary function parameters. The MDS program is safe and easy to practice. It should be considered as a mode for exercise prescription in the elderly. Keywords: Yoga; Elderly; Functional fitness; QoL

Quantifying Bone Marrow Fat Fraction and Iron by MRI for Distinguishing Aplastic Anemia from Myelodysplastic Syndromes

ROIs were manually drawn at the level containing the largest section of the left posterior superior ilium (A, B, D, E). The zoomed version of the ROIs showed in the upper left corner. (A, D) FF value of AA and MDS is 86.98% and 25.84%, respectively. (B, E) Iron content from R2*image is 117.91 and 147.98/second. (C, F) Histologic sections images of AA and MDS. Compared with MDS group (F), the number of fat cells (black arrow) in the AA group (C) was increased by Zeng et al. (1754–1760) image

BMSCs Regulate the Function of NK Cell By CD155/Tigit/CD226 Pathway in MDS Patients

Objective To analyze the immune status of MDS patients and evaluate its impact on prognosis.To investigate the specific mechanism of how mesenchymal stromal cells (MSCs) affecting immunity.Methods The study included 84 healthy controls and 84 inpatients with primary MDS, 22 patients with AML who were initially diagnosed in the Department of Hematology, General Hospital of Tianjin Medical University from January 2017 to July 2021.We collected flow cytometry(FCM) reports and clinical data. All MDS patients were stratified by IPSS-R for disease risk. According to IPSS-R classification, Very Low/Low and Int/High/Very high were divided into lower-risk MDS group and higher-risk MDS group, respectively.1. Statistical analysis of lymphocyte subset counts and cytokines in peripheral blood of different types of patients and healthy controls. 2. Correlation analysis between hemoglobin , platelets,neutrophil, blast cells, cytogenetics and lymphocyte counts was performed.3.We generated MSCs in BM samples from MDS patients and HCs.The expression of CD155 in MSCs and TIGIT/CD226 in lymphocytes were detected by FCM.Results 1.Percentage counts of NK cells in the HC group(19.94,95%CI 17.86-22.02，P<0.001)were significantly higher than other groups.The higher-risk group of MDS(11.51,95%CI 9.34-13.68) was obviously lower than the lower-risk group(16.91,95%CI 13.85-19,97)(P=0.006),close to the AML group(9.77,95%CI 6.33-13.20,Figure1).Meanwhile, the level of IFN-Ƴ in serum which is secreted mainly by NK cells was also the highest in the HC group(5.50,95%CI 3.74-7.26), followed by the MDS group and the lowest in the AML group(2.64,95%CI 1.91-3.37,Figure1).In addition, CD8+ T cells and CD19+ cells had similar results as NK cells, but there was no significant difference with CD3+ cells and CD4+ T cells.2.Significantly higher level of NK cells percentage counts were found in patients with hemoglobin ≥ 100 ×10 12/L compared to patients with lower hemoglobin(23.23,95%CI 17.26-29.21 and 15.77,95%CI 13.00-18.55,P=0.010). While the levels of NK cells were higher in patients with blast cells≤5% than in those with higher blast cells.(19.51,95%CI 16.09-22.93 and 14.65,95%CI 11.55-17.74,P=0.027). No significant difference was found between NK cell levels with regard to platelet and neutrophil counts.CD8+ T cells behaved similarly to NK cells. 3.The expression of CD155 on the surface of BMSCs was significantly higher in the MDS group than that in the HC group(P=0.001，Figure3).Moreover,MDS patients had higher expression of TIGIT(P=0.012) and lower expression of CD226(P=0.001,Figure4B) on the surface of NK cells in bone marrow than that in HCs.And CD8+ T cells also have similar results as NK cells.We analyzed the correlation between TIGIT of NK cells and CD155 of BMSCs, and found that they were negatively correlated（P=0.0297）, while CD226 was positively correlated with CD155（P=0.0706，Figure 4C）.Conclusions The decreased immune function in MDS patients is associated with prognosis.BMSCs are responsible for inducing an immune-suppressive microenvironment in MDS through CD155/TIGIT/CD226 pathway, the underlying mechanism remains to be further study. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Clonal Hematopoiesis in Normal Individuals Is Not Random and Likely Reflects Early MDS

Introduction: Clonal hematopoiesis, as determined by the presence of TET2, ASXL1, and DNMT3 mutation, is believed to be increasingly common in normal individuals as they age. This phenomenon is currently referred to as clonal hematopoiesis of indeterminate potential (CHIP). It has been reported that a few of these patients will develop myeloid neoplasms that begin as MDS and may evolve into AML. However, the clinical relevance of this particular subset of gene mutations (TET2, ASXL1, and DNMT3A) in healthy individuals is unclear as there are more than 50 genes that have been reported to be involved in myeloid neoplasms.FLT3 and NPM1 mutations have been reported in de novo AML patients and may occur as a secondary mutations on an MDS background. In order to clarify the significance of TET2, ASXL1, and DNMT3 in CHIP and their relationship to MDS and AML, we assessed the prevalence of TET2, ASXL1, and DNMT3 genes in patients with NPM1 and FLT3 mutations. In addition, since IDH1/2 gene are rarely mutated with TET2, we also studied the frequency of mutations of various myeloid genes in this group of patients.Methods: A total of 6390 consecutive bone marrow aspirate samples or peripheral blood samples submitted with clinical impression of AML, MDS, or MPN between January 2014 and mid 2017 were tested for mutations in myeloid genes using NGS. We used the TruSight Myeloid Panel (Illumina, San Diego, CA) for detecting missense mutations and fragment length analysis (FLA) for detecting ITD in FLT3 and large indels in CALR . DNA was extracted from samples using the QIAamp DNA Mini Kit. This NGS testing covers mutations in 54 myeloid-related genes. The average depth of sequencing was 10,000x.Results: In our database of consecutively tested patients, there were 311 patients with FLT3 mutations, 318 patients with NPM1 mutation, and 467 patients with IDH1/2 mutation. In addition, there were 308 patients diagnosed as MDS. All these patients tested for mutations in all 54 myeloid-related genes. The median age of patients in FLT3, MDS, IDH1/2, and NPM1 groups was 65, 75, 69, and 66, respectively. The top 4 most frequently mutated in genes in the FLT3 -positive patients, which present AML, were NPM1, DNMT3, WT1, and RUNX1 . In patients with NPM1 mutations, also representing AML, the top 4 most mutated genes were FLT3, IDH2, NRAS, and PTPN11 . In the IDH1/2 mutated patients, the top 4 mutated genes were DNMT3A, SRSF2, ASXL1, and NPM1 . As expected in the MDS group, the top 4 mutated genes were TET2, DNMT3A, ASXL1, and SRSF2 .Conclusions: The demonstration that mutations in TET2, DNMT3A, and ASXL1 are most common in patients presenting with MDS, as well as in CHIP, but not in patients presenting with acute myeloid neoplasms, suggests that CHIP likely represents early MDS. It is possible that mutations in these genes in few hematopoietic cells may not be adequate for manifestation of clinical MDS, but the presence of these mutations in large number of cells (high VAF) or the accumulation of mutations in additional genes is necessary for clinical disease. Therefore the clinical relevance of CHIP should always be considered in conjunction with other mutations and with VAF. [Display omitted] DisclosuresMa:NeoGenomics: Employment. De Dios:NeoGenomics: Employment. Funari:NeoGenomics: Employment. Sudarsanam:NeoGenomics: Employment. Jiang:NeoGenomics: Employment. Agersborg:NeoGenomics: Employment. Hummel:NeoGenomics: Employment. Blocker:NeoGenomics: Employment. Albitar:NeoGenomics: Employment.

Patients with Myelodysplastic Syndromes, Other Than Del(5)q Syndrome, Exhibiting Del(5)q Alone or Associated with One Additional Chromosomal Abnormality Have Comparable Probability and Duration of Response to Lenalidomide, with Patients Classified As Del(5)q Syndrome

Background: Lenalidomide (Len) has been approved for anemic patients with Del(5)q syndrome and induces high response rates. However, patients exhibiting Del(5)q abnormality, but not classified as Del(5)q syndrome or presenting additional abnormalities over Del(5)q are not usually treated with lenalidomide and the experience on these patient groups is limited.Aim: We investigated response rates and duration of response to Len in 67 MDS patients, exhibiting Del(5)q, but not classified as Del(5)q syndrome and compared their clinical and laboratory features with those of 120 patients with Del(5)q syndrome, treated with Len the same time period. Data were collected from the Hellenic (Greek) National MDS Registry database.Methods: Patients were classified in four groups: Group-1 (N=120), with the typical Del(5)q syndrome, Group-2 (N=25), exhibiting Del(5)q alone, but not fulfilling criteria for Del(5)q syndrome, Group-3 (N=22), with Del(5)q plus 1 additional cytogenetic abnormality and Group 4 (n=20), exhibiting Del(5)q plus >1 additional cytogenetic abnormalities. Results were expressed on an “intention to treat” basis. Patients were considered evaluable for response if they had completed at least 2 cycles of Len treatment.Results: Group-2 patients were significantly younger, compared to those of all the remaining Groups (p<0.05 for all comparisons) and the female to male ratio was higher in Group-1 (4.8 vs 2.1, 2.2 and 2.3 for Groups-2, -3 and -4, respectively). According to WHO classification patients of Group-2 were classified as RAEB-1 (n=16), RAEB-2 (n=6), CMML-D (n=1) and AML (n=2), patients of Group-3 as RA (n=7), RCMD (n=8), RAEB-1 (n=4), RAEB-2 (n=1) and AML (n=1) and patients of Group-4 as RCMD (n=5), RAEB-1 (n=5), RAEB-2 (n=8), CMML-P (n=1) and AML (n=1). Overall, 22 patients (Group-1: 11, Group-2: 1, Group-3: 5 and Group-4: 5) were not evaluable for response. Among 165 evaluable patients a complete hematological response was obtained by 83/109 (76.1%) patients from Group-1, 16/24 (66.7%) from Group-2, 9/17 (52.9%) from Group-3 and by 4/15 (26.7%) from Group-4. Partial response was achieved by 14, 4, 3 and 3 patients from each group, respectively, comprising overall response rates of 89%, 83.3%, 70.6% and 46.7%, respectively. Collectively, a favorable response was obtained by 26/38 (68.4%) patients with excess of blasts or AML in Groups-2 and -3 or by 32/41 (78%) patients from Groups -2 and -3. The median duration of response was 22.3 months in Group-1 and 17.3 months in Groups -2 and -3 (p: n.s.). Median duration of response was higher for patients without excess of blasts (21.6 vs 12.2 months, p<0.01). Complete cytogenetic response (CCyR) was confirmed in 28/56 patients from Group-1 and in 9/29 patients from the remaining Groups. CCyR was associated with higher MCV decrease and with reduction or disappearance of bone marrow biopsy-detected lymphoid nodules in 26/45 cases. Kaplan-Meier estimates for median overall survival following Len start was 67.6 (95% C.I. 43.4-91.8) months for Group-1, 43.7 (23.5-63.9) months for Group-2, 26.4 (22.1-30.6) months for Group-3 and 21.7 (12.7-30.7) months for Group-4 patients (p<0.001). In multiparametric analysis, factors associated with longer survival were achievement of complete hematological response (p<0.001), shorter interval from initial diagnosis to Len start (p<0.001), younger patient's age (p=0.01) and deeper MCV decrease at best response (p=0.05).Conclusion: Len may be highly effective, not only in patients with Del(5)q syndrome, but also in other MDS patients, exhibiting Del(5)q alone, or associated with 1 additional cytogenetic abnormality, even when excess of blasts is detected and might be considered as a treatment option in these MDS groups. [Display omitted] DisclosuresVassilakopoulos:Roche: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Celgene/GenesisPharma: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; BMS: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding. Katodritou:Janssen: Honoraria; Takeda: Honoraria, Research Funding; Amgen: Honoraria; Celgene: Honoraria, Research Funding. Pagoni:Gilead: Honoraria; Janssen: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; BMS: Consultancy, Honoraria. Angelopoulou:Janssen: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Gilead: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; BMS: Consultancy, Honoraria, Research Funding; Celgene/GenesisPharma: Consultancy, Honoraria, Research Funding. Labropoulou:Amgen: Honoraria; Celgene/GenesisPharma: Honoraria; Janssen: Honoraria. Terpos:Amgen: Honoraria, Other: SC member, Research Funding; Janssen: Honoraria, Research Funding; Genesis/Celgene: Honoraria, Other: DMC member, Research Funding; Takeda: Honoraria, Other: SC member; BMS: Honoraria; Abbvie: Honoraria; GSK: Honoraria.

A Comparative study between Decompressive Craniectomy with Multidural Stabs and Craniotomy with Durotomy in Acute Subdural Hematoma

Introduction: An acute subdural hematoma (SDH) is one of the most lethal of all head injuries. A good number of patients are encountered in Dhaka medical college and hospital with acute subdural hematoma. There are many modalities for the treatment of acute subdural hematoma, such as decompressive craniectomy with multidural stabs, craniotomy with durotomy. This comparative study was carried -out in the Department of Neurosurgery, Dhaka Medical College and Hospital, Dhaka, during Jan 2016 – Dec 2017, to compare the effect of the decompressive craniectomy with multidural stabs and craniotomy with durotomy in respect to the post operative GCS and GOS. As well as to assess and compare the post operative complications. The type of the study is prospective randomized controlled clinical trial.&#x0D; Materials and Methods: For this purpose, a total of 56 patients of both sex and any age with acute subdural hematoma attended in the above hospital were included in this study. Among them 28 patients underwent decompressive craniectomy with multidural stabs and rest 28 underwent craniotomy with durotomy. Data were collected and analyzed by different variables like age, sex, GCS on admission, GCS on 7th POD, GOS at one month and some complication like operative recurrence, seizure and mortality. Patients or attendants who refuse interview, who managed conservatively and patient who didn’t give consent, patients having severe respiratory distress and or shock, who came after 24 hours of trauma and who had no history of trauma were excluded from the study.&#x0D; Results: The following observations and results were obtained in this study. Nearly two third (63.0%) patients of DC MDS group and 18 (64.3%) in CT DT group age belonged to 21 to 40 years. Most (85.7%) of the patients of DC MDS group and 22 (78.6%) in CT DT group were male. Almost a half (46.4%) patient of DC MDS group and 12 (42.8%) patients of CT DT group GCS on admission belonged to 4 to 8 scale. The differences between two groups were not significant (p&gt;0.05). Nearly a half (46.4%) patient of DC MDS group and 6 (21.4%) patients of CT DT group GCS on 7th POD belonged to 9 to 12 scale. Mortality observed almost one third (32.1%) and 12 (42.9%) in patients of DC MDS group and CT DT group respectively. Regarding the GOS at one month follow up, it was observed 12 (42.9%) patients of DC MDS group and 2 (7.1%) patients of CT DT group had good recovery. About the relation between GCS (on admission) with GOS at one month, we saw that 10(37.7%) patient. GCS was ≥8 where GOS was 4+5 and compare to CT with DT group 9 (32.1%) patients. The difference was statistically significant (p&lt;0.05) and GCS (7th POD) with GOS, it was seen 15 (53.3%) patients GCS was e”8 where GOS was 4+5 and compare to CT with DT group 9 (32.1%) patients. About the complications 4 (14.3%) patients of DC MDS group had both surgical recurrence and seizures. 12 (42.9%) patients of CT DT group had seizures and 11 (39.3%) patients had surgical recurrence.&#x0D; Conclusion: Though mortality between these two groups was not significant, GCS on 7th POD and GOS on 30th POD were significantly better in decompressive craniectomy with multidural stabs group and operative recurrence and seizure was less in decompressive craniectomy with multidural stabs group and was statistically significant.&#x0D; Bang. J Neurosurgery 2021; 10(2): 192-200

Antibiotic self-medication and knowledge about antimicrobial resistance among medical and non-medical students of the University of Dodoma, Tanzania

Background: Antibiotic misuse and overuse are the major driving factors of antimicrobial resistance (AMR). High-income countries have been conscious of the problem for several years, while Tanzania shows the increasing trend of self-medication with antibacterial drugs. This study aimed to compare the antibiotics use practice and awareness about AMR among undergraduate students of the largest Tanzanian university. Methods and materials: Self-administered questionnaires were distributed among 2nd year medical (MDS) and non-medical students (non-MDS) at the University of Dodoma, Tanzania. A total of 356 respondent's answers were analyzed in the study. Statistical analysis was done by using MedCalc statistical software. p < 0.05 was considered statistically significant. Results: Non-MDS were more prone to use self-medication with antibiotics compared to MDS group: 46.2% and 35.1%, respectively (OR 1.58, 95%CI: 1.0 to 2.5). Among them, 45.5% of non-MDS and 49.2% of MDS reported an inability to visit a doctor as the main reason for the self-treatment. Moreover, non-MDS replied that they did not attend the doctor for financial reasons (OR 4.64, 95%CI: 2.05 to 10.46). Most students relied on a previous experience to choose medicine type: 76.3% MDS and 57.6% non-MDS (OR 2.36, 95%CI: 1.09 to 5.13). The next most common approach, notably popular among Tanzanian students, was to follow Pharmacist's recommendations: 22% MDS and 24.2% non-MDS. All surveyed participants demonstrated a high level of awareness regarding the antibiotic indications. However, less than half of MDS (46.2%) knew about AMR, and only 19.1% of non-MDS were familiar with the problem (OR 3.63, 95%CI 2.22 to 5.95). Furthermore, 48.8% of MD and 27.3% of non-MD students did not finish an entire course of antibiotics (OR 2.54, 95%CI 1.58 to 4.09) and stopped taking a drug mainly when the symptoms of disease disappeared. Analysis of antibiotic trends showed Ampicillin, Amoxicillin and others from Penicillin group to be the most popular among Tanzanian undergraduates. However, many students confused antibiotics, pain killers and antiallergic medicines. Conclusion: Our findings indicated the high rate of self-medication with antibiotics and AMR illiteracy among undergraduate students that proved the necessity of educational programs, antibiotic use guidance and the control of sales of antibiotics in the country.