Megaloblastic Anemia Patients Research Articles

Changes and clinical significance of erythrocyte lifespan in megaloblastic anemia

Objective: To investigate the lifespan of erythrocytes in megaloblastic anemia (MA) patients. Methods: A prospective cohort study analysis. Clinical data from 42 MA patients who were newly diagnosed at the Department of Hematology, Lanzhou University Second Hospital from January 2021 to August 2021 were analyzed, as were control data from 24 healthy volunteers acquired during the same period. The carbon monoxide breath test was used to measure erythrocyte lifespan, and correlations between erythrocyte lifespan and laboratory test indexes before and after treatment were calculated. Statistical analysis included the t-test and Pearson correlation. Results: The mean erythrocyte lifespan in the 42 newly diagnosed MA patients was (49.05±41.60) d, which was significantly shorter than that in the healthy control group [(104.13±42.62) d; t=5.13,P=0.001]. In a vitamin B12-deficient subset of MA patients the mean erythrocyte lifespan was (30.09±15.14) d, and in a folic acid-deficient subgroup it was (72.00±51.44) d, and the difference between these two MA subsets was significant (t=3.73, P=0.001). The mean erythrocyte lifespan after MA treatment was (101.28±33.02) d, which differed significantly from that before MA treatment (t=4.72, P=0.001). In MA patients erythrocyte lifespan was positively correlated with hemoglobin concentration (r=0.373), and negatively correlated with total bilirubin level (r=-0.425), indirect bilirubin level (r=-0.431), and lactate dehydrogenase level (r=-0.504) (all P<0.05). Conclusions: Erythrocyte lifespan was shortened in MA patients, and there was a significant difference between a vitamin B12-deficient group and a folic acid-deficient group. After treatment the erythrocyte lifespan can return to normal. Erythrocyte lifespan is expected to become an informative index for the diagnosis and treatment of MA.

Mechanism of megaloblastic anemia combined with hemolysis

ABSTRACT Megaloblastic anemia (MA) patients often exhibit hemolysis, but it is not clear whether there are other hemolytic mechanisms in addition to intramedullary hemolysis. We retrospectively analyzed the clinical characteristics of 124 MA patients, measured erythrocyte physical parameters in two patients with hemolysis and one healthy volunteer by atomic force microscopy, and measured 18F-FDG uptake in one MA patient with hemolysis. In multivariate analysis, hemolysis was associated with mean corpuscular volume (MCV) and indirect bilirubin. A receiver operating characteristic curve analysis, with sensitivity of 83.1% and specificity of 68.7%, suggested that the MCV cutoff value that predicts hemolysis is 116.4 fL. Hb was negatively correlated with MCV in the hemolysis group (r = −0.317, P = 0.007) but not in the nonhemolysis group. The erythrocyte peak-valley value, average cell surface roughness and surface area in the MA patients with hemolysis were significantly lower than those in controls (P < 0.05). 18F-FDG uptake by the liver and spleen was diffuse and increased in MA patients undergoing hemolysis. MA combined with extramedullary hemolysis could be caused by macrophages removing mechanically damaged erythrocytes and the retention of erythrocytes with decreased deformability when blood circulates through narrow spaces in the liver and spleen.

Schistocytosis in megaloblastic anemia masquerading peripheral blood picture as maha (microangiopathic hemolytic anemia) and decrease mean corpuscular volume

Severe megaloblastic anemia presenting with hemolytic anemia, thrombocytopenia and schistocytosis resemble microangiopathic hemolytic anemia. Presence of scistocytes falsely decreases the Mean corpuscular volume. Toestimate the degree of schistocytosis in megaloblastic anemia with hemolytic blood picture and its correlation with the lowering of MCV. 60 cases of megaloblastic anemia.30 patients of megaloblastic anemia with hemolysis(cases) and 30 patients without hemolysis(clinical control)were taken. Complete blood count, Peripheral smear examination, schistocyte count, serum bilirubin, LDH, Homocystine was done. Mean age of clinical control group was 22 where as that of cases was 14.1 years. Mean serum homocysteine level was increased in both clinical control group and cases. In our study of 30 megaloblastic anemia patients with hemolysis (case), 29 had &amp;#62;1% of schistocytes in their peripheral blood picture. There is a positive correlation between percentage of schistocytes and homocysteine and a negative correlation between MCV and homocysteine. There is a statistically significant negative correlation existing between percentage of schistocytes and MCV.Presence of scistocytes in peripheral blood should not be always considered as MAHA, and lowering of MCV should not be confused as microcytic hypochromic anemia. Presence of macroovalocytes and hypersegmented neutrophils can be a clue for diagnosis of megaloblastic anemia with hemolysis in such type of cases.

Etiologyical Analysis of 133 Patients with Elderly Macrocytic Anemia and Diagnostic Significance of Laboratory Tests

To study the etiology of macrocytic anemia in elderly patients and to evaluate the diagnostic significance of laborotory tests. 133 elderly macrocytic anemia patients, whose age>60 years old, hemoglobin<100 g/L, mean red cell volume(MCV)>100 fL, and bone marrow cell test was performed, and these patients were grouped according to diseases, and the bilirubin, lactate dehydrogenase, folic acid, vit B12 and serum ferritin were tested, then the results of tests were compared and analyzed. The majority of the cases were diagnosed as megaloblastic anemia (MA), myelodysplasia syndrome (MDS), acute leukemia/multiple myeloma (AL/MM) and hemolytic anemia (HA). Usually HA was a simple anemia, while others were accompanied by decrease of other 1 or 2 series. HA patients were often with significant high level of well volume (MCV), red cell distribution width(RDW), reticulocytes (RC) and indirect bilirubin (IBIL) (P<0.01). However, MA patients were often with high level of LDH. Serum ferritin (SF) level was significantly higher in both MDS and AL/MM groups (P<0.01). Common causes of macrocytic anemia in elderly patients are MA, MDS, AL/MM and HA. The combination detection of MCV, RDW, RC, LDH, IBIL and SF contributes to enhancing the accuracy of diagnosis.

Value of MCV/RDW Combined with Reticulocyte Parameters in Differential Diagnosis of Anemia Diseases

To evaluate the value of mean corpuscular volume/RBC distribution width (MCV/RDW) combined with reticulocyte parameters in differential diagnosis of aplastic anemia (AA), myelodysplastic syndrome (MDS), megaloblastic anemia (MA) and hemolytic anemia (HA) in order to provide some laboratorial evidence for clinical doctors in first diagnosis of these diseases. The data of MCV/RDW and reticulocyte parameters of AA, MDS, MA and HA patients from January 1 of 2011 to August 31 of 2014 were retrospectively collected in West China Hospital of Sichuan University. And 158 healthy unrelated individuals with age-, sex-matched were collected as controls. The value of MCV/RDW and reticulocyte parameters in differentiating diagnosis of above mentioned 4 kinds of anemia diseases was assessed. ROC analysis was used to determine the cutoff value of MCV/RDW and the reticulocyte parameters were performed in differentiating diagnosis of AA and MDS. The average values of MCV/RDW of 158 AA patients (79 acute AA patients and 79 chronic AA patients), 107 MDS patients, 13 MA patients and 81 HA patients increased in variable degrees as compared with the controls, and there was statistical difference between them, the MCV/RDW value of acute AA patients was obviously less than that of other patients. In the 4 kinds of anemia diseases, the reticulocyte absolute count in acute AA patients was the lowest, that of chronic AA, MA and MDS patients was higher, and that of HA patients was highest. The ratio of low fluorescent reticulocyte decreased, and the ratio of moderate and high fluorescent reticulocytes increased in the 4 kinds of anemia diseases, as compared to controls. The difference was statistically significant. The analysis of differential diagnosis of chronic AA and MDS showed that RDW-SD could differentiate the chronic AA from MDS. The area under the curve (AUC) of RDW-SD was 0.76 (P < 0.01). The cutoff value of RDW-SD was 22.75fl. The sensitivity and specificity of RDW-SD for differential of chronic AA and MDS was 49.5% and 98.7%, respectively. MCV/RDW and reticulocyte parameters can be used as the laboratorial differential diagnostic indicators for AA, MDS, MA and HA diseases.

Autophagy level of bone marrow mononuclear cells in patients with myelodysplastic syndromes

目的研究骨髓增生异常综合征（MDS）患者骨髓单个核细胞（BMMNC）自噬水平的变化，探讨细胞自噬在MDS中的作用。方法以38例MDS患者为实验组，以26例巨幼细胞贫血患者为对照组，抽取骨髓并分离BMMNC。应用透射电镜观察自噬情况；采用单丹（磺）酰戊二胺（MDC）染色法检测自噬泡水平；采用RT-PCR、免疫荧光技术及蛋白免疫印迹法检测微管相关蛋白轻链3（LC3）、Beclin1 mRNA及蛋白的表达水平。结果MDS患者BMMNC内易见自噬泡；MDC染色显示MDS患者BMMNC含自噬泡细胞数量显著高于对照组[（9.75±2.63）%对（2.90±0.89）%，P<0.05]；MDS患者BMMNC LC3阳性细胞比例显著高于对照组[（6.13±1.03）%对（1.50±0.58）%，P<0.05]。低危/中危-1 MDS患者BMMNC Beclin1及LC3A mRNA表达水平显著高于对照组（3.61±3.02对1.55±1.03，P<0.05；6.56±3.97对1.21±0.95，P<0.05）；低危/中危-1 MDS患者BMMNC自噬负性调控基因-哺乳动物雷帕霉素靶点（mTOR）mRNA水平显著低于对照组（0.39±0.37对1.50±1.03，P<0.05）；中危-2/高危MDS患者BMMNC Beclin1、LC3和mTOR mRNA表达水平与对照组比较差异均无统计学意义（P值均>0.05）。蛋白免疫印迹法显示低危/中危-1患者BMMNC Beclin1蛋白表达（1.257±0.197）高于对照组（0.528±0.086）及中危-2/高危（0.622±0.118），差异均有统计学意义（P<0.05）。中危-2/高危组和对照组比较差异无统计学意义（P>0.05）。结论低危/中危-1 MDS患者BMMNC自噬水平升高，中危-2/高危MDS组患者BMMNC自噬水平无明显改变，自噬对MDS患者可能起保护性作用，自噬水平相对不足可能与MDS的进展有关。

Identification of four SLC19A2 mutations in four Chinese thiamine responsive megaloblastic anemia patients without diabetes

Identification of four SLC19A2 mutations in four Chinese thiamine responsive megaloblastic anemia patients without diabetes

Expression characteristics of differentiation antigens on granulocytes in patients with megaloblastic anemia

This study was aimed to explore the change characteristics of cell differentiation antigen (CD) on bone marrow (BM) granulocytes in patients,with megaloblastic anemia (MA). In combination with BM cell morphology, hemogram, level of blood serum folic acid, level of Vit B(12), cell genetics and biological examination data, the BM granulocytes differentiation antigens in 13 patients with MA were detected by flow cyto metry and analyzed retrospectively, in order to summarize the variation characteristics of CD13, CD33 and CD15 expressed on myeloid cells in patient with MA, including forward scatter light (FSC) and side scatter light (SSC) signal intensity, then these findings were compared with that in normal healthy persons. The results showed that the expression rates of CD13, CD15 and CD33 on granulocytic in patients with MA and normal healthy persons were (44.53 ± 16)%, (96.16 ± 2.67)%, (80.81 ± 14.71)% and (62.33 ± 11.02)%, (99.53 ± 0.46)%, (70.00 ± 7.81)% respectively, in which the expression rate of CD13 and CD15 in patients with MA decreased (P < 0.01), while the expression rate of CD33 increased (P < 0.01). The mean fluorescence intensity (MFI) of CD13, CD15, CD33, SSC and FSC in MA patients and normal healthy persons were 3.39 ± 1.41, 14.29 ± 6.59, 1.95 ± 0.94, 478.78 ± 70.43, 633.46 ± 75.53 and 5.12 ± 1.15, 20.67 ± 5.13, 1.04 ± 0.17, 332.00 ± 38.16, 537.00 ± 16.70 respectively, in which the MFI of CD13 and CD15 on granulocytes in MA patients decreased (P < 0.01),while the MFI of FSC,SSC and CD33 increased (P < 0.01 and P < 0.05). It is concluded that not only the morphology of BM granulocytes in patents with MA shows dysmaturity, but the expressing feature of differentiation antigens on BM granulocytes in MA patients also displays dysmaturity.These findings will contribute to the clinical diagnosis of MA patients.

Vitamin B12-homocysteine interaction and the efficacy of B12 therapy in relation to anemia and neurological disease in a North Indian population

Background: Vitamin B12 deficiency leads to myriad hematological and neurological changes leading to megaloblastic anemia and disorders of the nervous system like subacute combined degeneration of the spinal cord, cognitive impairment, peripheral neuropathy, etc. In this case series, an attempt was made to investigate the neurological manifestations of patients with diagnosed vitamin B12-deficient megaloblastic anemia, determine the cause of vitamin deficiency in these cases, and monitor their response to vitamin B12 therapy. Materials and Methods: The study was conducted in a tertiary care referral teaching hospital in New Delhi during a period of over 2 years from August 2007 to December 2009. Eighty-five consecutive patients with megaloblastic anemia (MA) due to vitamin B12 deficiency were screened and a total of 28 patients with neurological and psychiatric symptoms were selected for the study. In addition to bone marrow examination, blood counts, red cell indices, and biochemical tests, assays of serum vitamin B-12, folate, and homocysteine were done. Neuroimaging by magnetic resonance studies and nerve conduction studies (NCS) was done where clinically applicable. Follow-up was done at least twice at 3 months and 6 months to assess clinical, neurological, and hematological response to vitamin B12 therapy. Results: The primary study group of MA with B12 deficiency had 85 patients. Of these, only 28 (32.9%) patients had neurological and/or psychiatric features. There were 18 male and 10 female subjects (M: F ratio being 1.8:1). The mean age was 32.80 years with a range of 18-68 years. Though the majority of MA patients (77.6%) were vegetarian, a good 22% consumed a nonvegetarian diet. Neurological manifestations included peripheral neuropathy in 14 (50%), myelopathy in 2 (7.14%), myeloneuropathy in 2 (7.14%), acute confusional state in 4 (14.28%), and cognitive impairment in 20 (71.42%) patients. Mood disturbances, anxiety, and irritability were also seen. NCS was done in 18 cases and showed demyelinating peripheral neuropathy in 12 cases and both axonal and demyelinating changes in two cases. Magnetic resonance imaging (MRI) showed four cases with the classical demyelination on spinal MRI with white matter hyper intense lesions in the posterior tracts on T2 weighted images. At 6-month follow-up with B12 therapy, 10/14 patients with neuropathy recovered completely, 2/14 had partial recovery, and 2 had poor recovery. While the cases of myelopathy had only partial recovery at 6 months, all four cases with acute confusional state recovered completely in response to B12 therapy. Conclusion: The electrodiagnostic and neuroimaging changes in vitamin B12 deficiency are consistent with focal demyelination of white matter tracts in the brain, the spinal cord, and peripheral nerves. In this case series of B12 deficiency MA, we found demyelinating peripheral neuropathy to be the predominant presenting neurological disease. However, cognitive impairment with behavioral changes was found to be common on neurological and psychiatric examination. Many of these features were found to be reversible with B12 therapy.

Nutritional megaloblastic anemia during childhood: Demographical, clinical and laboratory features of 134 patients from southeastern part of Turkey

Summary Background & aims Nutritional megaloblastic anemia is a common disease among young children in the developing and underdeveloped countries. The aim of this study was to examine the laboratory and clinical characteristics of megaloblastic anemia patients diagnosed at Pediatric Hematology Unit of Dicle University Hospital. Methods Between March 2002 and February 2007, a total of 665 patients with anemia were examined and 134 (20%) cases were diagnosed with nutritional megaloblastic anemia. Medical records of 134 patients (74 male, 55.2%; 60 female, 44.8%) were evaluated retrospectively. Results Of these, 126 (94%) were from low socio-economical backgrounds and 110 (82%) were younger than 2 years of age. The most common reasons for admission were paleness (48%), high fever (29%), diarrhea (23%), weakness (21%), irritability (21%), failure to thrive (18%), vomiting (16%), and neurological findings such as tremor (16%) and inability to walk (2%). The diagnosis was based on physical examination, peripheral blood smear, complete blood count, vitamin B12, and folate levels in all patients. Bone marrow aspiration was made only in 41 patients. This sample population contained a high proportion of young children from lower socio-economical backgrounds (82% Conclusions The diagnosis of nutritional megaloblastic anemia should always be borne in mind in young children presenting with anemia in the developing and underdeveloped countries.

Gene expression profiling of the bone marrow mononuclear cells from patients with myelodysplastic syndrome

The gene expression pattern of bone marrow mononuclear cells (BMNCs) from 10 patients with myelodysplastic syndrome (MDS) was studied by two-color cDNA microarray techniques. To confirm the microarray results, a semiquantitative RT-PCR was performed to analyze gene expression in fifty additional MDS patients. Ninety-five genes were shown to be abnormally expressed in at least five MDS patients compared to normal controls, involving cell growth and differentiation regulation, cell cycle control, signaling and redox; such as thrombospondin 1, phosphatase and tensin homolog, MAD, DNA-damage-inducible transcript 3 (DDIT3), ets variant gene 1 (ETV1), and G1 to S phase transition 1. CD36 was also revealed up-regulated in 4 cases. MDS patients in early and advanced stages could be clustered into two distinct groups by hierarchical clustering, wherein a case with isolated thrombocytopenia and other RA patients were clustered into two subgroups. Consistent expression patterns of 3/5 (60%) genes were confirmed by semiquantitative RT-PCR. Further analysis showed the different transcript levels of RNAHP, DDIT3 in patients with MDS in different stages, AML, and normal controls. Meanwhile, the different significance of RNAHP and ETV1 expression was revealed between RA and untypical anaplastic anemia, iron deficiency anemia, and megaloblastic anemia patients. We propose that the technology of microarray may reveal the intrinsic molecular features and the expression levels of RNAHP, DDIT3, and ETV1 may provide useful markers for the diagnosis of MDS.

Circulating and Local Bone Marrow Renin-angiotensin System in Leukemic Hematopoiesis: Preliminary Evidences

Until last the decade, the renin-angiotensin system (RAS) was considered as a circulating endocrine system. It is now known that there are local RASs in many tissues. It has also recently been hypothesized that there exists a local bone marrow (BM) RAS with paracrine/autocrine pathobiological functions. The aim of this study was to detect BM and peripheral blood levels of the essential RAS components in normal and leukemic hematopoiesis.Concentrations of renin and angiotensin-converting enzyme (ACE) were assayed in BM aspirates and in simultaneously drawn peripheral blood samples of 16 pre-chemotherapy leukemic and 10 post-treatment megaloblastic anemia patients with normal blood counts, as controls.In the leukemia group, the ACE concentration was found to be significantly higher in the BM (38±6.2 U/l) than in the peripheral blood (29.5±5.3 U/l), (p=0.029). In the leukemia group, although the BM renin concentration was higher than the peripheral blood levels (21.3±8.3 vs. 18.6±6.2 U/l), this difference was not statistically significant (p=0.196). In the control group, mean BM renin levels were insignificantly lower than in the peripheral blood (8.6±3 vs. 12.1±4.6 pg/ml) (p=0.059). In the leukemia group, serum ACE levels positively correlated with BM and peripheral blood blast percentages (p<0.05). Serum LDH level (p<0.01), BM blast (p<0.05) and peripheral blast percentages (p<0.01) were inversely correlated with serum potassium in the leukemia group.The results of this study can be considered as the preliminary evidence supporting the hypothesis of the presence of a local BM RAS. Further, molecular biologic and immunohistochemical studies are needed to shed light on this important subject. A better understanding of the interrelationships of RAS and hematopoiesis will bring new insights into the pathobiology and even novel therapies for such neoplastic diseases.

Sensitivity to Thiamine Deficiency in Cultured Human Cells is Dependent on Cell Type and is Enhanced in Cells From Thiamine-Responsive Megaloblastic Anemia Patients

To address tissue-specific variation in sensitivity to thiamine deficiency, three human cell types were grown in medium with various thiamine concentrations. The activity of a cytosolic and a mitochondrial thiamine diphosphate-dependent enzyme was examined. Each cell type displayed a unique response to thiamine depletion with respect to α-ketoglutarate dehydrogenase and transketolase activity and to inhibition of cell growth. Loss of α-ketoglutarate dehydrogenase activity was similar in lymphoblasts and fibroblasts, whereas loss of activity in neuroblastoma cells was significantly more resistant to thiamine depletion. Transketolase activity in neuroblastoma cells was only moderately resistant to thiamine depletion, with the activity in fibroblasts being the most and in lymphoblasts the least resistant. Total transketolase activity was 33% higher in fibroblasts than in lymphoblasts and neuroblastoma cells, indicating a differential requirement for production and maintenance of transketolase activity in this cell type. Compared with normal lymphoblasts, those derived from patients with thiamine-responsive megaloblastic anemia were 100 to 1000 times more sensitive to thiamine depletion. Although fibroblasts from these patients also demonstrated a 1000-fold increase in sensitivity with respect to transketolase activity, α-ketoglutarate dehydrogenase activity demonstrated no enhanced sensitivity. The results indicate a complex, cell-type dependent regulation of intracellular pools of thiamine and its phosphorylated derivatives in response to fluctuating extracellular thiamine levels.