Peripheral Blood Hemogram Research Articles

Environmental pollutant ENU induced leukemic NF-kB signaling amelioration by Eclipta alba in murine model

ABSTRACT Exposure to N-nitroso compounds (NOCs) in our environment via pesticides, tobacco, and smoked meat can be potentially carcinogenic. The induction of N-N’ ethylnitrosourea (ENU), a genotoxic NOC, leads to leukemogenesis. The study aimed to explore the ameliorating effect of the Ayurvedic herb Eclipta alba on the bone marrow cells of ENU-induced leukemic mice. Eclipta alba is investigated for its anti-cancer effect on various cell lines, but never on haematological malignant models. Theefficacy of the extract was explored on leukemia by changes in body weight, survivability, peripheral blood hemogram, bone marrow cytological, histological, and cell culture studies pre-and post-treatment. The treated group revealed significant immunomodulation of the expressional profile of NF-kB family and IL-1β in marrow cells, by flow-cytometry, and immunofluorescence study. Through our experimental endeavour we depicted the cellular mechanism, signaling modality and tried to establish the anti-cancer potency of Eclipta alba on ENU-induced leukemia.

Effect of ionizing radiation on the occupational health of radiologists

Objective To analyze peripheral blood hemogram, lymphocyte micronucleus and chromosomal aberrations of radiologists, so as to provide basis for occupational protection and health monitoring of radiologists. Methods Lymphocyte micronucleus, chromosome and blood hemogram analysis were performed on 127 radiologists who received health examinations in 2015, 2017 and 2019, and they were assigned to the radiation group. In addition, 133 medical staff with no history of radiation exposure were selected as the control group. Results The micronucleus rate and chromosome aberration rate of the radiation group were higher than those of the control group, and the white blood cell and platelet counts were lower than those of the control group, both of which were statistically significant ( P < 0.05). The total number of white blood cells in peripheral blood of 127 radiologists decreased gradually with the increase of exposure time to ionizing radiation, and the chromosome aberration rate increased gradually, all of which had statistical significance ( P < 0.05). The rate of chromosomal aberration was higher in radiologists with damage work age of more than 20 years than in the low-work age group, and there was no statistical significance between different damage work age ( P > 0.05). The chromosome aberration rate of nuclear medicine and interventional therapy was higher than that of other types, with statistical significance ( P < 0.05). Conclusion Long-term exposure to low-dose ionizing radiation can reduce the total number of white blood cells and increase the chromosome aberration rate of radiologists. It is necessary to strengthen the protective measures for radiologists to reduce the degree of ionizing radiation damage, especially to strengthen the occupational protection for radiologists in nuclear medicine and interventional therapy. 摘要： 目的 分析放射工作者外周血象、淋巴细胞微核及染色体畸变情况, 为放射工作者职业防护和健康监测提供依 据。 方法 对 2015 年、2017 年和 2019 年连续 3 次接受健康检查的 127 名放射工作者进行淋巴细胞微核、染色体及血 象分析, 将其设为放射组。另外选取 133 名无射线接触史的医务人员设为对照组; 结果 放射组中淋巴细胞微核率和 染色体畸变率高于对照组, 白细胞和血小板计数低于对照组, 均具有统计学意义 ( P < 0.05)。127 名放射工作者外周血 白细胞总数随着接触电离辐射时间的增长逐渐降低, 染色体畸变率逐渐增加, 均具有统计学意义 ( P < 0.05)。损害工龄 大于 20 年的放射工作者染色体畸变率高于低工龄组, 不同损害工龄之间比较无统计学意义 ( P > 0.05)。核医学与介入 治疗工种染色体畸变率高于其他工种, 具有统计学意义 ( P < 0.05)。 结论 长时间接触低剂量电离辐射可使放射工作 者白细胞总数降低和淋巴细胞染色体畸变率增加, 应加强放射工作者防护措施以备降低电离辐射损伤程度, 特别要加 强核医学和介入治疗放射工作人员的职业防护。

Biological activity of 4-ethynyl-, 4-oxy-, 4-butoxypropylpyperidine and azaheterocyclic compounds

The new chemical series of compounds is promising for the search new effective immunostimulating drugs. Acute toxicity and the effect on peripheral blood hemogram parameters and on the subpopulation composition of lymphocytes were determined in the compounds (piperidine-containing derivatives): 1-(3-n-butoxypropyl)-piperidine-3-methyl-4-spiro-5'-imidazolidine-2',4'-dione,1-(2-ethoxyethyl)-4-carboxy-4-hydroxypiperidine, bis[1-(2-ethoxyethyl)-4-ethynyl-4-hydroxypiperidine) dihydrochloride and, 1:1 complexes of β-cyclodextrin with 1-(2-ethoxyethyl)-4-(2-ethoxyethoxyl)-4-ethynylpiperidine, 1-(2-ethoxyethyl)-4-(2-methoxyethoxyl)-4-ethynylpiperidine, 7-(2-methoxyethyl)-2,2-dimethyl-3-thia-7-azabicyclo[3.3.1]nonane or 3-(2-morpholinoethyl)-7-(3-isopropoxypropyl) diazabicyclo[3.3.1] nonane with β-cyclodextrin. Compound ˗ 1-(3-n-butoxypropyl)piperidine-3-methyl-4-spiro-5'-imidazolidine-2',4'-dione, exceeds the immunostimulating activity of the comparison drug of levamizole by 3,1 times and has low toxicity.

Expression and Function of miR-99a-5p in Bone Marrow of Patients with MDS

To detect the expression of miR-99a-5p in myelodysplastic syndrome (MDS), to predict the target genes and to analyze its function by using bioinformatics. The expression levels of bone marrow miR-99a-5p in MDS patients were detected by qRT-PCR, and the correlation of miR-99a-5p expression with clinical pathological characteristics, percentage of marrow blasts , chromosome karyotype and peripheral blood hemogram were analyzed. The target genes of miR-99a-5p were predicted by Targetscan, Miranda and Microcosm, and the intersection of the predicted results of 3 softwares was used as a potential target gene for miR-99a-5p. The expression level of bone marrow miR-99a-5p in MDS patients was significantly higher than that of healthy controls (P<0.01); According to the prognostic score of IPSS, MDS patients were divided into relatively low risk group (including low risk group and intermediate risk group 1) and relatively high risk group (including intermediate risk group 2 and high risk group). Analysis showed that the expression level of bone marrow miR-99a-5p in relatively low risk group was 2.40 times higher than that in healthy control group (P<0.01), the expression level of bone marrow miR-99a-5p in relatively high risk group was 6.66 times higher than that in healthy control group (P<0.01), the expression level of miR-99a-5p in relatively high risk group was 2.80 times higher than that in the relatively low risk group ( P<0.01) ; the expression level of bone marrow miR-99a-5p in t-MDS group was 6.35 times higher than that in healthy control group (P<0.001). There was a significant positive correlation between the expression level of miR-99a-5p and the percentage of marrow blasts (P<0.01), but the expression of miR-99a-5p did not correlate with chromosome karyotype and peripheral blood hemogram; In this study it was obtained that ST5 might participate in pathological mechanism of MDS by bioinformatic analyses and references. The expression levels of miR-99a-5p is up-regulated in MDS patients, and its expression showed a rising tendency which may be involved in the pathogenesis of MDS by regulating target gene ST5.

Genomic insult oriented mitochondrial instability and proliferative hindrance in the bone marrow of aplastic mice including stem/progenitor population

Aplastic anemia is the bone marrow failure condition characterized by the development of hypocellularity in both marrow and peripheral blood compartments. Anti-tumor chemotherapeutic agents often exert secondary effect on hematopoietic system leading to aplastic anemia by marrow failure. The precise mechanisms behind the marrow ablative effects of the drugs remain yet to be established. The present study holds a mechanistic approach to unveil the mystery. Aplastic anemia was generated in mice with the administration of busulfan and cyclophosphamide followed by the characterization of the disease with peripheral blood hemogram, histopathological and cytochemical examinations of bone marrow. To gain deep knowledge about the molecular mechanisms of the hematopoietic disruption, cytotoxicity assay, DNA damage measurement, apoptosis study, replicative senescence analysis, redox balance study, mitochondrial membrane potential change assessment, flowcytometric expressional analysis of p21, p53, ATM, Chk-2, Necdin, Gfi-1, c-myc, KU-80 and Sod-2 were done with marrow hematopoietic stem/ progenitor cells (HSPCs). Severe blood pancytopenia and marrow hypocellularity was found in aplastic mice. Proliferative hindrance and apoptosis of marrow cells were identified as the cause behind the hematopoietic catastrophe. The genotoxic effects of the drugs triggered chromatin damage and induced replicative senescence in aplastic HSPCs by upregulating p21 in a p53 independent manner. Moreover, accumulation of genomic insults also caused apoptotic elimination of marrow cells due to disruption of mitochondrial membrane potential by generating redox imbalance. The study established the underlying mechanisms behind hematopoietic disruption during drug induced marrow aplasia. Outcome of the study may be helpful in successful designing of therapeutic strategies for the disease concerned.

Deregulation of vital mitotic kinase-phosphatase signaling in hematopoietic stem/progenitor compartment leads to cellular catastrophe in experimental aplastic anemia.

Aplastic anemia, the paradigm of bone marrow failure, is characterized by pancytopenic peripheral blood and hypoplastic bone marrow. Among various etiologies, inappropriate use of DNA alkylating drugs like cyclophosphamide and busulfan often causes the manifestation of the dreadful disease. Cell cycle impairment in marrow hematopoietic stem/progenitor compartment together with cellular apoptosis has been recognized as culpable factors behind aplastic pathophysiologies. However, the intricate molecular mechanisms remain unrevealed till date. In the present study, we have dealt with the mechanistic intervention of the disease by peripheral blood hemogram, bone marrow histopathology, cytopathology, hematopoietic kinetic study, scanning electron microscopy, DNA damage assessment and flowcytometric analysis of cellular proliferation and apoptosis in hematopoietic stem/progenitor cell (HSPC) rich marrow compartment using busulfan and cyclophosphamidemediated mouse model. To unveil the molecular mechanisms behind aplastic pathophysiology, we further investigated the role of some crucial mitotic and apoptotic regulators like Protein kinase-B (PKB), Gsk-3β, Cyclin-D1, PP2A, Cdc25c, Plk-1, Aurora kinase-A, Chk-1 regarding the hematopoietic catastrophe. Our observations revealed that the alteration of PKB-GSK-3β axis, Plk-1, and Aurora kinase-A expressions in HSPC compartment due to DNA damage response was associated with the proliferative impairment and apoptosis during aplastic anemia. The study established the correlation between the accumulation of DNA damage and alteration of the mentioned molecules in aplastic HSPCs that lead to the hematopoietic catastrophe. We anticipate that our findings will be beneficial for developing better therapeutic strategies for the dreadful disease concerned.

Alteration of classical and hematopoiesis specific p53 pathway in the bone marrow hematopoietic stem/progenitor compartment facilitates leukemia progression in experimental mice

Downregulation of p53 is associated with most of the neoplasms, however it claims additional significance for hematopoietic malignancy due to its supplementary role during hematopoiesis. Apart from the classical role as tumor suppressor, p53 during steady state hematopoiesis is associated with the maintenance of quiescent cell population in bone marrow by upregulating necdin (Ndn) and Gfi-1. We felt, it is necessary to delineate its attribution towards malignant conversion of hematopoietic system during leukemogenesis from all the possible angles. The present study deals with the characterization of N-N′ Ethylnitrosourea (ENU) induced mouse model of leukemia by peripheral blood hemogram, bone marrow cytology, histology, cytochemical staining (MPO) and scanning electron microscopic study. We further investigated the alteration of conventional and hematopoiesis specific p53 pathways by flowcytometric expressional analysis of ATM, Chk-2, p53, p21, Ndn, Gfi-1 and Tie-2. The disruption of classical p53 pathway was observed in leukemic hematopoietic stem/progenitor population which involved downregulation of ATM, Chk-2, p53 and p21. Moreover, the expressional decline of Ndn and Gfi-1 hinted towards the mechanism of hindrance of hematopoietic quiescency in leukemic bone marrow. Increased expression of Tie-2 due to reverse correlation with p53 was found to be responsible for pathological angiogenesis in bone marrow together with increased blast burden in bone marrow during leukemia. The study presents the mechanistic scenario of the alteration of both classical as well as hematopoiesis specific p53 pathways in HSPC compartment triggering leukemic pathophysiology.

Pathophysiological Scenario of Hematopoietic Disorders: A Comparative Study of Aplastic Anemia, Myelodysplastic Syndrome and Leukemia in Experimental Animals

The conversion of physiology to pathophysiology in hematological disorders viz: aplastic anemia, myelodysplastic syndrome (MDS) and leukemia in murine models was the subject of study in the present programme. Peripheral blood hemogram, spleno-somatic index, bone marrow smear study, cytochemical staining of marrow, cell release kinetics study during marrow explants culture, hematopoietic niche assessment, chromosomal aberration study, plasma membrane stability study of marrow cells, lysosomal membrane and mitochondrial membrane stability study and innate immune parameters were performed in the aplastic anemia, leukemia and MDS mouse model. In bone marrow aplasia, peripheral blood pancytopenia, marrow hypocellularity, decreased marrow cellular viability, deterioration of bone marrow hematopoiesis as well as hematopoietic microenvironment and extramedullary hematopoiesis were noticed. In addition, disruption of mitochondrial and lysosomal membrane integrity along with reduction of innate immune parameters were found in the hematopoietic suppressed condition. Surprisingly, no noticeable chromosomal aberration was found in the aplastic condition. Ineffective marrow hematopoiesis together with the disruption of hematopoietic microenvironment was observed in MDS. Also, extramedullary hematopoiesis, increased marrow cellular death, chromosomal aberration and loss of innate immunity were the common events. During leukemia, the number of functionally and structurally immature cells in the peripheral blood and bone marrow was increased together with malignant conversion of hematopoietic cells in the presence of malignancy supportive stromal microenvironment. Chromosomal aberration, decrease of cell mediated immunity with least mitochondrial apoptotic damage were also found in leukemic condition as well.

Sodium orthovanadate (vanadate), a potent mitigator of radiation-induced damage to the hematopoietic system in mice.

Previous in vitro and in vivo studies have shown that sodium orthovanadate (vanadate), an inorganic vanadium compound, could effectively suppress radiation-induced p53-mediated apoptosis via both transcription-dependent and transcription-independent pathways. As a potent radiation protector administered at a dose of 20 mg/kg body weight (20 mg/kg) prior to total body irradiation (TBI) by intra-peritoneal (ip) injection, it completely protected mice from hematopoietic syndrome and partially from gastrointestinal syndrome. In the present study, radiation mitigation effects from vanadate were investigated by ip injection of vanadate after TBI in mice. Results showed that a single administration of vanadate at a dose of 20 mg/kg markedly improved the 30-day survival rate and the peripheral blood hemogram, relieved bone marrow aplasia and decreased occurrence of the bone marrow micronucleated erythrocytes in the surviving animals. The dose reduction factor was 1.2 when a single dose of 20 mg/kg was administered 15 min after TBI in mice using the 30-day survival test as the endpoint. Results also showed that either doubling the vanadate dose (40 mg/kg) in a single administration or continuing the vanadate treatment (after a single administration at 20 mg/kg) from the following day at a dose of 5 mg/kg per day for 4 consecutive days further significantly improved the efficacy for rescuing bone marrow failure in the 30-day survival test. Taken together, these findings indicate that vanadate would be a potent mitigator suppressing the acute lethality (hematopoietic syndrome) and minimizing the detrimental effects (anhematopoiesis and delayed genotoxic effects) induced by TBI in mice.

Hemopoiesis in pregnant beagles following low-dose total-body irradiation and surgery.

Hemopoietic perturbations were observed in gravid beagles subjected to whole-body ionizing radiation during midpregnancy followed by the additional trauma of surgery, with alterations in peripheral blood hemogram parameters, bone marrow morphology, and concentration of marrow-derived granulocytic- and erythrocytic-committed stem cells. However, the trauma of surgery combined with irradiation had no apparent effects on the contralateral uterine horn with the remaining pups, with no indications of miscarriage, absorption of pups, or other complications of pregnancy.