Proliferation Of Bone Marrow Cells Research Articles

Immunostimulatory Effect of Postbiotics Prepared from Phellinus linteus Mycelial Submerged Culture via Activation of Spleen and Peyer's Patch in C3H/HeN Mice.

Medicinal mushrooms are an important natural resource promoting health benefits. Herein, Phellinus linteus mycelia were prepared under submerged cultivation, the mycelium-containing culture broth was extracted as a whole to obtain the postbiotic materials (PLME), and its effect on the immune system was evaluated in normal C3H/HeN mice. Oral administration of PLME for 4 weeks was well tolerated and safe. In the PLME-administered groups, in addition to the production of immunostimulatory cytokines, such as interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6), the mitogenic activity was significantly increased. PLME administration also significantly increased the levels of serum immunoglobulin G (IgG) and IgA in the small intestinal fluid and Peyer’s patches and enhanced Peyer’s patch-mediated bone marrow cell proliferation activity and cytokine production (IL-2, IL-6, and IFN-γ). Histomorphometric analyses showed an increase in immune cells in the spleen and small intestinal tissues of mice administered PLME, supporting the rationale for its immune system activation. PLME mainly contained neutral sugar (969.1 mg/g), comprising primarily of glucose as a monosaccharide unit. The β-glucan content was 88.5 mg/g. Data suggest that PLME effectively promote immune function by stimulating the systemic immune system through the spleen and intestinal immune tissues. PLME can thus be developed as a functional ingredient to enhance immune functions.

Modulation of local and systemic immune responses by fermented garlic extract

Fermented garlic (FG), prepared by fermenting garlic with <em>Bacillus subtilis</em> for 3‑15 days at 20‑40˚C has been linked to immune‑boosting properties. In the present study, FG was utilized to investigate its immunomodulatory effects using RAW264.7 cells, as well as BALB/c and C3H/HeN mice. Nitric oxide (NO) and pro‑inflammatory cytokine levels were quantified in RAW264.7 cells treated with or without FG (0.016, 0.08, 0.4, 2 and 10 mg/ml), garlic extract (GE) (0.016, 0.08, 0.4, 2 and 10 mg/ml) and lipopolysaccharide (1 µg/ml). Peritoneal macrophages and splenocytes were obtained from BALB/c mice for <em>ex vivo</em> analyses. MTT and enzyme‑linked immunosorbent assays were used to determine splenocyte proliferation, natural killer (NK) cell activity and cytokine production. The intestinal immunostimulatory <em>in vitro</em> effects of FG were also investigated. In a concentration‑dependent manner, FG treatment markedly accelerated the proliferation of RAW264.7 cells, murine peritoneal macrophages and splenocytes. Moreover, FG initiated macrophage activation by significantly increasing the NO, tumor necrosis factor‑α, interleukin (IL)‑12 and IL‑6 levels in RAW264.7 cells and peritoneal macrophages. In contrast to FG, GE promoted RAW264.7 cell proliferation at lower concentrations and toxicity at higher concentrations, while no detectable NO synthesis was observed at the concentrations tested. Likewise, the FG extract at larger concentrations fortified the intestinal immune system and improved bone marrow cell proliferation by significantly elevating IL‑6 and granulocyte‑macrophage colony‑stimulating factor production in splenocytes and Peyer's patch cells of mice. In addition, the NK cell activities of splenocytes from FG‑treated mice at lower doses against YAC‑1 cells were significantly increased, which must have been fostered and activated by interferon‑γ generated by splenocytes. On the whole, these findings provide relevant <em>in vitro</em> and <em>ex vivo</em> information that describes the mechanisms through which FG can strengthen a compromised immune system by influencing local and systemic gut immune responses directly or indirectly.

P1004: EFFECT AND MOLECULAR MECHANISM OF TQ05105, A NOVEL SMALL MOLECULE INHIBITOR OF JAK2 IN MYELOPROLIFERATIVE NEOPLASM

Background: The JAK2V617F gene mutation leads to upregulation of the JAK/STAT signaling pathway, which causes excessive bone marrow cell proliferation, elevated inflammatory cytokines, constitutional symptoms and shortened survival in myeloproliferative neoplasms (MPN) patients. The JAK1/2 kinase inhibitor ruxolitinib can dramatically improve disease-related symptoms, but side effect such as anemia and thrombocytopenia limit its use in a substantial proportion of patients. Several JAK2 kinase inhibitor III Clinical trials have yielded unsatisfactory outcomes recently. TQ05105 is a new small molecule inhibitor that targets JAK2 kinase. Aims: We aim to investigate the effect and molecular mechanism of TQ05105 on MPN cell lines and mouse models. Methods: We systematically measured the function of TQ05105 in MPN cell lines, including UKE1 and SET2 cells, by performing proliferation, apoptosis and colony-forming assays. RT-PCR and inflammatory factor multi-assay kit was used to detect the secretion of inflammatory factors such as TNF-α, IL-1β, MCP-1 in mRNA level and protein level respectively. CFU-E and BFU-E assays were performed on MPN cell lines and primary cells. In vivo, TQ05105 was evaluated in JAK2V617F Jak2V617F transplant mouse model and MPL515 retroviral mouse model. Mice were orally administrated with placebo, 25 mg/kg (bid) and 50 mg/kg (bid) every day for 4 weeks. Flow cytometry analysis was performed to determine the proportion of different hematopoietic progenitor cell populations. In addition, HE staining and Gomori staining was used to detect the changes in the spleen and bone marrow. Results: In UKE1 and SET2 cell lines, TQ05105 can inhibit the proliferation and induced cell apoptosis, notably reduce the expression of cytokines, including TNF-α, IL-1 and MCP-1. According to Western blot results, the phosphorylation of JAK/STAT signaling pathway associated proteins JAK2, STAT3 and STAT5 were significantly suppressed by TQ05105. Furthermore, compared to the control group, TQ05105 can strongly limit the ability of UKE1 cell lines, SET2 cell lines and primary cells from MPN patients (PMF, PV, ET) to form BFU-E and CFU-GM. TQ05105 can striking reduce mouse spleen volume, along with favorable impact on leukocyte and erythrocyte counts in Jak2V617F transplant mouse model. The proportion of stem and progenitor cells, especially CMPs and MEPs, was considerably reduced in mice after treatment with TQ05105. HE staining showed remarkably Spleen response such as recovering structure, reduced infiltration of erythrocytes and suppressed proliferation of megakaryocytes. MPLW515L retroviral mouse model recapitulated the feature of myelofibrosis, application of TQ05105 in this model resulted in reduction of the spleen volume, extramedullary hematopoiesis, leukocyte and platelets count, bone marrow fibrosis and decreased fraction of CMPs and MEPs. Summary/Conclusion: TQ05105, as a novel small molecule JAK2 inhibitor, can inhibit cell proliferation, induce cell apoptosis and decrease inflammatory cytokines by affecting the JAK/STAT signaling pathway. In vivo experiments further proved that TQ05105 can reduce spleen size and improve disease-related symptoms of MPN, indicating that it is a promising therapeutic drug for MPN.

Effect of Salvia purpurea cav. on the Proliferation of Hematopoietic Cells In Vitro

In Mexico, some plants of the Salvia genus are employed against digestive problems, circulatory problems, infections, and some hematological alterations. We evaluated the effect of Salvia purpurea Cav. (Labiateae) on the proliferation of mouse hematopoietic cells. The extracts were prepared by consecutive maceration with hexane, dichloromethane, methanol, and water from three mixtures of the aerial structures of the plant: flower; flower–leaf, and flower–leaf–stem. Their hematopoietic capacity was determined in cultures of mouse bone marrow and spleen utilizing concentrations of 1, 10, and 100 µg/mL of each extract. The 100-µg/mL concentration of the flower extracts obtained with hexane, dichloromethane, and water established the proliferation of bone-marrow cells. The 10-µg/mL concentration of the hexanic and dichloromethanic extracts reduced cellularity by 34 ad 33%, respectively (p &lt;0.005). Likewise, the 10-µg/mL concentration of the hexanic extract reduced the cell count by 30% (p &lt;0.025). The 100-µg/mL concentration of the four flower–leaf extracts increased the concentration from 135–174%. Of the 36 extracts tested in bone marrow, 14 were stimulants, 19 were cytostatic, and three, cytotoxic. The remaining extracts increased the cellular concentration from 127 to 199.6%. In spleen cultures, the 100-µg/mL concentration of the flower–leaf–stem, and the hexanic and methanolic extracts increased the cellular concentration. The 100-µg/mL concentration of the flower–leaf extracts obtained with dichloromethane and water gave rise to 4.0 and 3.4 increments in the cell count, respectively. The phytochemical study revealed the presence of phenolic and saponin compounds. &#x0D; Keywords: Hematopoietic activity, Genus Salvia, Salvia purpurea Cav, Bone marrow, Spleen.

Peculiar features of bone marrow cell proliferation in Djungarian hamsters with genetic disorders under thiotepa effect

The paper aims to examine the proliferation of bone marrow cell pool in Djungarian hamsters and the subsequent restoration of their genetic stability after the action of thiotepa (TT). The study involved 36 animals, of which 16 were in the control group (injected with 0.25 ml of physiological solution), and 20 in the experimental group (0.25 ml of thiotepa at a dose of 1.5 mg per 1 kg of body weight). The maximum number of cells with CA amounting to 30.0% was observed 13 hours after TT injection (p≤0.05 between the control and experimental groups) and rapidly declined to 5.7% over subsequent periods by the 37th hour of the experiment (p≤0.05). The results suggest that the restoration of cell pool genetic stability is largely associated with the cell selection mechanisms, which confers an advantage over cell proliferation without chromosome anomalies.

Two birds with one stone: YQSSF regulates both proliferation and apoptosis of bone marrow cells to relieve chemotherapy-induced myelosuppression

Ethnopharmacological relevanceYiqi Shengsui formula (YQSSF) is a commonly used formula to treat chemotherapy-induced myelosuppression, but little is known about its therapeutic mechanisms. Aim of this studyThis study aims to examine the effect of YQSSF in treating myelosuppression and explore its mechanism. Materials and methodsA myelosuppression BALB/c mouse model was established by intraperitoneal (i.p.) injection of cyclophosphamide (CTX). The efficacy of YQSSF in alleviating chemotherapy-induced myelosuppression was evaluated by blood cell count, immune organ (thymus, spleen, liver) index, bone marrow nucleated cell (BMNC) count and histopathological analysis of bone marrow and spleen. Then, ultra-performance liquid chromatograph quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was performed to analyze the ingredients of YQSSF extract. Key effects and potential mechanism of YQSSF extract in alleviating myelosuppression were predicted by network pharmacology method. Finally, cell cycle and TUNEL staining of bone marrow cells was detected to verify the key effects, and RT-qPCR or Western blotting were performed to measure the gene and protein expressions of the effect targets respectively to confirm the predicted mechanism of YQSSF for myelosuppression. ResultsYQSSF up-regulated the number of peripheral blood leukocytes and BMNC, reduced spleen index and liver index, improved the pathological morphology of bone marrow and spleen. A total of 40 ingredients were isolated from YQSSF extract using UPLC-Q/TOF-MS analysis. Network pharmacology revealed that YQSSF regulated both proliferation and apoptosis to alleviate myelosuppression. Finally, YQSSF decreased G0/G1 ratio, increased the proportion of bone marrow cells in S phase and proliferation index (PI), and reduced apoptotic cells in femur bone marrow. RT-qPCR and Western blotting showed that YQSSF up-regulated the expression levels of CDK4, CDK6, CyclinB1, c-Myc and Bcl-2, as well as down-regulated the expression levels of Cyt-c, Fas, Caspase-8/3 and p53. ConclusionsYQSSF promotes the proliferation and inhibits the apoptosis of bone marrow cells to relieve chemotherapy-induced myelosuppression.

АНАЛИЗ ПОЯВЛЕНИЯ МИКРОЯДЕР В ЭРИТРОЦИТАХ И АКТИВНОСТИ ПРОЛИФЕРАЦИИ КЛЕТОК КОСТНОГО МОЗГА ПОСЛЕ ПРОЛОНГИРОВАННОГО ОБЛУЧЕНИЯ МЫШЕЙ БЫСТРЫМИ НЕЙТРОНАМИ В НИЗКИХ ДОЗАХ

Purpose: To analyze the level of cytogenetic damage and the activity of bone marrow cells proliferation in C57BL/6 mice after prolonged fast neutrons low dose irradiation at 10–500 mGy. &#x0D; Material and methods: Male C57BL/6 mice at the age of 7–8 and 16 weeks were used in the experiments. Irradiation was carried out on an OR-M installation in the field of fast neutrons and gamma quanta using five Pu(α,n)Be radionuclide sources with a high fast neutron yield at a dose rate of 2.13 mGy/h. The frequency of polychromatophilic (PCE) and normochromic (NCE) erythrocytes with micronuclei (MN) and the ratio of PCE and NCE were analyzed using light microscopy after cytochemical staining of the bone marrow cells of control and irradiated mice. The proliferation activity of bone marrow cells was determined by the number of Ki-67+-cells. The parameters of the cell cycle and the level of apoptosis were studied after DNA staining with DAPI using flow cytometry. Statistical processing of the results was carried out according to the Student’s method using the computer program Origin.&#x0D; Results: It was found that prolonged irradiation of mice with fast neutrons at a low dose rate (2.13 mGy/h) at doses from 10 to 500 mGy after 24 h led to statistically significant increase in the frequency of PCE with MN at all studied doses. No dose dependence of this parameter was observed in the studied range. The increase in the frequency of PCE with MN at a dose of 500 mGy was prolonged and persisted for at least 72 h. A significant increase in the frequency of NCE with MN 24 h after irradiation was found only at a dose of 500 mGy, which persisted up to 48 h. At this dose, there was also a decrease in the number of nucleated cells in the bone marrow 24 – 72 h after exposure, a decrease in the number of Ki-67+-cells 24 h after irradiation of mice, a block of the cell cycle in the G2/M phase, and a decrease of cells in the G0/G1 phase, but after 48 h, there were no disturbances in the cell cycle. &#x0D; Conclusion: It has been shown that after a single total prolonged irradiation of mice at low doses (10–500 mGy), when analyzing the frequency of PCE with MN, cytogenetic damage is recorded in the bone marrow, which indicates the genetic danger of exposure to even such low levels of fast neutron irradiation. A decrease in Ki67+ cells and cell cycle arrest at the G2/M phase were found only after irradiation of mice at a dose of 500 mGy and only 24 h after exposure, while the number of nucleated cells in the bone marrow at this dose was reduced, at least to 72 h.

Reduction of Cell Proliferation by Acute C2H6O Exposure.

Simple SummaryAlcoholic beverages and acetaldehyde formed during their metabolism are carcinogenic to humans. Alcohol drinking may affect bone marrow stem cell niche, suppressing physiological hematopoiesis and ultimately reducing the organism’s capacity to fight against cancer, infections, and to promote tissue regeneration. To elucidate in vivo the cellular mechanisms associated with alcohol intake toxicity, we used a mouse model in which proliferating cells produce the firefly’s light-emitting protein. In this animal, alcohol exposure transiently “turns off the light”, indicating a negative effect on cell proliferation in the bone marrow and spleen. Pharmacological treatment with substances interfering with ethanol metabolism, reducing acetaldehyde production, partially restores the physiological cell proliferation rate. Over 560 million people worldwide have increased susceptibility to acetaldehyde toxicity and 4% of cancer deaths are attributable to alcohol. Our model might provide a suitable tool to further investigate in vivo the effects of alcohol metabolism and aldehydes production on carcinogenesis.Endogenous acetaldehyde production from the metabolism of ingested alcohol exposes hematopoietic progenitor cells to increased genotoxic risk. To develop possible therapeutic strategies to prevent or reverse alcohol abuse effects, it would be critical to determine the temporal progression of acute ethanol toxicity on progenitor cell numbers and proliferative status. We followed the variation of the cell proliferation rate in bone marrow and spleen in response to acute ethanol intoxication in the MITO-Luc mouse, in which NF-Y-dependent cell proliferation can be assessed in vivo by non-invasive bioluminescent imaging. One week after ethanol administration, bioluminescent signals in bone marrow and spleen decreased below the level corresponding to physiological proliferation, and they progressively resumed to pre-treatment values in approximately 4 weeks. Boosting acetaldehyde catabolism by administration of an aldehyde dehydrogenase activity activator or administration of polyphenols with antioxidant activity partially restored bone marrow cells’ physiological proliferation. These results indicate that in this mouse model, bioluminescent alteration reflects the reduction of the physiological proliferation rate of bone marrow progenitor cells due to the toxic effect of aldehydes generated by alcohol oxidation. In summary, this study presents a novel view of the impact of acute alcohol intake on bone marrow cell proliferation in vivo.

Effect of 50-Hz Magnetic Fields on Argyrophilic Nucleolar Organizer Regions in Bone Marrow Cells

Background: Exposure to an increasing amount of extremely low-frequency magnetic fields (MFs) causes some adverse effects. Considering a direct association between the sizes and numbers of argyrophilic nucleolar organizer regions (AgNORs) and cell proliferation, this study was conducted to evaluate the effect of 50-Hz MFs on the AgNORs of bone marrow (BM) cells. Methods: Eighty male rats were randomly divided into four exposed groups and one control group. The exposed groups received 50-Hz MFs with magnetic flux densities of 1, 100, 500, and 2000 µT two hours a day for two months. BM cells were aspirated from sacrificed rats’ femoral bones, smeared on glass slides, and then stained with silver nitrate for AgNORs. The area (AA), length (AL), and number (AN) of AgNORs were calculated by a microscope equipped with a camera and Scion Image software in 100 BM cells of each rat. The mean of AA, AL, and AN was computed for each group. Results: AA, AL, and AN significantly reduced in the 1 µT group compared with 2000 µT and control groups. Eventually, there was a nonlinear relationship between the effect of 50-Hz MFs and magnetic flux densities. Conclusion: Overall, 50-Hz MFs with a magnetic flux density of 1 µT reduced AgNORs in BM cells. However, 100, 500, and 2000 µT did not affect AgNORs. Therefore, 50-Hz MFs with low density may suppress BM cell proliferation.

Advanced glycosylation end products inhibit the proliferation of bone-marrow stromal cells through activating MAPK pathway

BackgroundThe purpose of present study was to explore the mechanism of nuclear factor-kappa B (NF-κB), phosphatidylinositol 3-kinase (PI3K)/protein kinase B(PKB/Akt) and mitogen-activated protein kinase (MAPK) signaling pathways after intervention of advanced glycosylation end products (AGEs) on rat bone-marrow stromal cells (BMSCs).MethodsPrepare and identify AGEs. BMSCs were isolated from 16 SD rats and cultured with different concentration of AGEs. Cell viability was detected by cell counting kit-8 (CCK-8). BMSCs were cultured with AGEs (0.25 mg/ml) for 30 min, 12 h, 24 h, 72 h and 120 h. In addition, BMSCs were cultured with AGEs, AGEs + JNK inhibitor and AGEs + P38 inhibitor for 24 h and 48 h, respectively. Western blotting and RT-PCR were used to determine the protein and mRNA expression levels, respectively.ResultsCell viability of BMSCs was significantly correlated with concentration and effect time of AGEs (P < 0.05), and the most appropriate concentration was 0.25 mg/ml. AGEs stimulation significantly increased the protein expression levels of NF-κB p65, JNK, p38 (P < 0.05), decreased IκB (P < 0.05), but had no effect on the protein expression of Akt in BMSCs (P > 0.05). At the mRNA level, JNK and p38 inhibitors significantly reduced the levels of NF-κB p65, p38 and JNK, increased IκB (P > 0.05), but had no effect on Akt in BMSCs (P > 0.05). At the protein level, JNK and p38 inhibitors notably decreased the expression of NF-κB p65, p38, p-JNK, P-IκB and JNK (P < 0.001), and increased IκB (P < 0.05).ConclusionAdvanced glycosylation end products can inhibit the proliferation of bone-marrow stromal cells through activating MAPK pathway.

Promotion of Bone Regeneration Using Bioinspired PLGA/MH/ECM Scaffold Combined with Bioactive PDRN.

Current approaches of biomaterials for the repair of critical-sized bone defects still require immense effort to overcome numerous obstacles. The biodegradable polymer-based scaffolds have been required to expand further function for bone tissue engineering. Poly(lactic-co-glycolic) acid (PLGA) is one of the most common biopolymers owing to its biodegradability for tissue regenerations. However, there are major clinical challenges that the byproducts of the PLGA cause an acidic environment of implanting site. The critical processes in bone repair are osteogenesis, angiogenesis, and inhibition of excessive osteoclastogenesis. In this study, the porous PLGA (P) scaffold was combined with magnesium hydroxide (MH, M) and bone-extracellular matrix (bECM, E) to improve anti-inflammatory ability and osteoconductivity. Additionally, the bioactive polydeoxyribonucleotide (PDRN, P) was additionally incorporated in the existing PME scaffold. The prepared PMEP scaffold has pro-osteogenic and pro-angiogenic effects and inhibition of osteoclast due to the PDRN, which interacts with the adenosine A2A receptor agonist that up-regulates expression of vascular endothelial growth factor (VEGF) and down-regulates inflammatory cytokines. The PMEP scaffold has superior biological properties for human bone-marrow mesenchymal stem cells (hBMSCs) adhesion, proliferation, and osteogenic differentiation in vitro. Moreover, the gene expressions related to osteogenesis and angiogenesis of hBMSCs increased and the inflammatory factors decreased on the PMEP scaffold. In conclusion, it provides a promising strategy and clinical potential candidate for bone tissue regeneration and repairing bone defects.

Bone marrow cells are differentiated into MDSCs by BCC-Ex through down-regulating the expression of CXCR4 and activating STAT3 signalling pathway.

Studies showed that the increase of myeloid‐derived suppressor cells (MDSCs) in tumour microenvironment is closely related to the resistant treatment and poor prognosis of metastatic breast cancer. However, the effect of tumour‐derived exosomes on MDSCs and its mechanism are not clear. Here, we reported that breast cancer cells (4T1)‐secreted exosomes (BCC‐Ex) were able to differentiate bone marrow cells into MDSCs and significantly inhibited the proliferation of T lymphocytes to provide an immunosuppressive microenvironment for cancer cells in vivo and in vitro. The number of MDSCs in bone marrow and spleen of 4T1 tumour‐bearing mice and BCC‐Ex infused mice was significantly higher than that of normal mice, whereas the number of T lymphocytes in spleen was significantly decreased. In addition, BCC‐Ex markedly promoted the differentiation of MDSCs from bone marrow cells or bone marrow cells derived macrophages, seen as the increased expressions of MDSCs‐related functional proteins Arginase‐1 (Arg‐1) and inducible nitric oxide synthase (iNOS). Furthermore, BCC‐Ex significantly down‐regulated the expressions of chemokine receptor CXCR4 and markedly up‐regulated the levels of inflammatory cytokines IL‐6 and IL‐10 in bone marrow cells and macrophages and remarkably inhibited the division and proliferation of T cells. Importantly, CXCR4 agonist, CXCL12, could reverse the function of BCC‐Ex, indicating that BCC‐Ex‐induced MDSCs might be dependent on the down‐regulation of CXCR4. Western blot showed that BCC‐Ex significantly promoted the phosphorylation of STAT3 in bone marrow cells, resulting in the inhibitions of the proliferation and apoptosis of bone marrow cells, and the aggravation of the differentiation of bone marrow cells into MDSCs.

Multifocal Multisystem Langerhans Cell Histiocytosis in an Adult Female Atypically Presenting With Multiple Abdominopelvic Abscesses

Introduction: Langerhans Cell Histiocytosis (LCH) is a condition of malignant clonal proliferation of myeloid bone marrow cells that more commonly affects children than adults. Furthermore, its presentation remains variable ranging from single organ to multisystem involvement including bone, skin, lymph, liver, spleen, lung, and central nervous system. We report an unusual case of multisystem LCH in an adult female presenting with multiple soft tissue abscesses. Case: We report a case of a 38-year-old female with a past medical history of polysubstance abuse, type 2 diabetes, polycystic ovarian syndrome, and isolated central diabetes insipidus who had multiple hospitalizations for recurrent soft tissue abscesses treated with incision and drainage and antibiotics. Imaging studies revealed multiple osteolytic lesions involving the bilateral iliac crests, acetabulum and femur, as well as an iliopsoas abscess. Given her prior history of isolated central diabetes insipidus, the possibility of LCH as the cause was entertained. Histological evaluation performed on an inguinal soft tissue sample stained positive for CD1a and S100, and a formal diagnosis of Langerhans Cell Histiocytosis (LCH) was made. The patient was treated with chemotherapy with good results. Conclusion: This report presents a rare and unusual case of adult onset multisystem LCH involving bone, skin, lymph, and central nervous system presenting with recurrent large abdominopelvic abscesses. These abscesses may represent a rare and unrecognized form of soft tissue involvement of LCH.

The Effect of Artemisia Dracunculus L. on Mitotic Index in Bone Marrow and Spleen Cells of Mice: In Vivo Study

Artemisia dracunculus L. (tarragon) is a rich source of herbal remedies with antioxidant and anti-inflammatoryproperties. In the present study, the proliferation of bone marrow and spleen cells of mice was evaluated afterextraction of Artemisia dracunculus with 80% of ethanol. Two doses of the extract (500mg/kg body weightand 1000mg/kg body weight) were given to the mice for seven successive days. On day eight, mice weresacrificed and cells from bone marrow and spleen were collected; mitotic index was calculated and resultswere compared with that of methotrexate at a dose 20mg/kg body weight ( positive control ) and distilledwater ( negative control). The results showed that the dose 500mg/kg body weight of Artemisia dracunculusextract caused significant increase in mitotic index in both bone marrow and spleen cells of mice whencompared with negative control. While, the dose 1000mg/kg body weight of Artemisia dracunculus extractcaused significant decrease in mitotic index in both bone marrow and spleen cells of mice when comparedwith negative control.

Bioactivity-guided investigation for isolation and immunoregulatory potential of polysaccharides from Dendrobium chrysotoxum stems

A new immunoregulatory polysaccharide (DCP-W4) with a relative molecular mass of 1.37 × 104 Da was separated and purified from the stems of Dendrobium chrysotoxum under the guidance of immunoregulatory activity assay based on the small intestinal Peyer’s patch-mediated proliferation of bone marrow cells. Analysis of physicochemical features showed that DCP-W4 was a glucomannan consisting of glucose and mannose in a molar ratio of 3.0 : 1.0. Oral administration of mice suggested that DCP-W4 could significantly influence the body’s immune activity, which was evidenced by its ability to change the indexes of immune organs including spleen and thymus, the proliferation of bone marrow cells mediated by the supernatant of Peyer’s patch cells, the balance of Th1/Th2 through effectively regulating the secretion and ratio of IFN-γ and IL-4, the proportion and differentiation status of different immune cells including T-cells, dendritic cells and macrophages, and the production of sIgA as well as β-defensins and mucin-2 in the small intestine or in the spleen of mice. These results indicated that DCP-W4 was a polysaccharide with a hopeful potential for the modulation of intestine immune system and systemic immune system.

Metabolism and chromatin: A dynamic duo that regulates development and ageing

Bone-marrow mesenchymal stem cell (BM-MSC) proliferation and lineage commitment are under the coordinated control of metabolism and epigenetics; the MSC niche contains low oxygen, which is an important determinant of the cellular metabolic state. In turn, metabolism drives stem cell fate decisions via alterations of the chromatin landscape. Due to the fundamental role of BM-MSCs in the development of adipose tissue, bones and cartilage, age-associated changes in metabolism and the epigenome perturb the balance between stem cell proliferation and differentiation leading to stem cell depletion, fat accumulation and bone-quality related diseases. Therefore, understanding the dynamics of the metabolism-chromatin interplay is crucial for maintaining the stem cell pool and delaying the development and progression of ageing. This review summarizes the current knowledge on the role of metabolism in stem cell identity and highlights the impact of the metabolic inputs on the epigenome, with regards to stemness and pluripotency.

Morphofunctional parameters of bone marrow of mice under using of antihypertensive agent

Due to the increase in cardiovascular disease, it is urgent to research new more effective and safe drugs and their combinations. Candesartan cilexetil is an angiotensin II receptor antagonist. It is known that candesartan cilexetil has a number of side effects, affecting the antioxidant system. Therefore, we studied the natural antioxidant resveratrol to prevent the cytotoxic effect. Goal. To study the effect of candesartan cilexetil and resveratrol in different doses on the number of apoptotic and micronucleated cells and cell cycle parameters in mice. Methods. Male C57Bl/6 mice were used for the experiment. Animals were received intragastrically candesartan cilexetil and resveratrol in different doses for 7 weeks. Evaluation of the effects of substances on cytogenetic and cytokinetic parameters of bone marrow was conducted by flow cytometry. Results. It was found that candesartan cilexetil at 1.5 mg/kg dose was cytotoxic. Using candesartan with resveratrol was safer for cells of bone marrow. Natural resveratrol reduced in combination the adverse effects of candesartan cilexetil. Test complex of substances stimulated cell proliferation in bone marrow of mice. Conclusions. New combination of candesartan cilexetil and resveratrol increased proliferation of cells and was not cytotoxic.

Autologous micrografts from the palatal mucosa for bone regeneration in calvarial defects in rats: a radiological and histological analysis

BackgroundThe application of dental implants is often restricted by bone volume. In such cases, bone grafts are required, although bone graft materials have some disadvantages. Therefore, other effective approaches are needed. Our previous study showed that the autologous micrograft, a dissociated cell suspension made out of palatal connective tissue grafts, promoted bone-marrow cell proliferation and differentiation under osteogenic conditions. In this study, we aimed to evaluate the effects of dissociated soft-tissue suspensions relevant to bone regeneration in animal model.Material and methodsTwelve-week-old male Wistar rats were used in the study. Defects were created in rat calvaria, and were filled with hydrogel containing either dissociated soft-tissue suspension (test) or sucrose (control). The new bone formation was evaluated at 1 and 2 weeks after surgery (n = 16) by radiological and histological analysis.ResultsThe conducted radiological analysis showed that the new bone volume was significantly greater in the dissociated soft-tissue suspension group. This finding was further confirmed by the conducted histological analysis.ConclusionsThe dissociated mucosa tissue suspension enhanced bone regeneration in vivo; thus, it is a promising potential method to aid the successful application for bone augmentation in the implant practice.

Immunostimulatory Effects of Live Lactobacillus sakei K040706 on the CYP-Induced Immunosuppression Mouse Model

Our previous studies have shown that heat-killed Lactobacillus sakei K040706 exerts immunostimulatory and anti-inflammatory activities in macrophages, cyclophosphamide (CYP)-treated mice, and dextran sulfate sodium–induced colitis mice. However, the immunostimulatory effects of live Lactobacillus sakei K040706 (live K040706) against CYP-induced immunosuppression and its underlying molecular mechanisms remain unknown. Therefore, we investigated the immunostimulatory effects of live K040706 (108 or 109 colony forming unit (CFU)/day, p.o.) in CYP-induced immunosuppressed mice. Oral administration of live K040706 prevented the CYP-induced decreases in body weight, thymus index, natural killer (NK) cell activity, T and B cell proliferation, and cytokine (interferon (IFN)-γ, interleukin (IL)-2, and IL-12) production. The administration of live K040706 also exerted positive effects on the gut microbiota of CYP-induced mice, resulting in a microbiota composition similar to that of normal mice. Moreover, live K040706 significantly enhanced IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) production in the splenocytes and Peyer’s patch (PP) cells of mice and increased bone marrow (BM) cell proliferation. Taken together, our data indicate that live K040706 may effectively accelerate recovery from CYP-induced immunosuppression, leading to activation of the immune system. Therefore, live K040706 may serve as a potential immunomodulatory agent against immunosuppression.

Abstract 16007: Myeloid-specific Deletion in Lipid Phosphate Phosphatase3 (plpp3) Increases Cardiac Inflammation

Introduction and Hypothesis: Acute myocardial infarction (AMI) is the leading cause of morbidity and mortality worldwide. The innate immune response plays a major role in cardiac remodeling after AMI. Lysophosphatidic acid (LPA), produced by autotaxin (ATX) and degraded by Lipid phosphate phosphatase 3 (LPP3), regulates monocytosis and promotes inflammation. However, the role of LPP3 in post-AMI inflammation is not understood. Here, we investigated the possible role of Myeloid-specific Plpp3 KO mice in cardiac and systemic inflammation and resulting in adverse cardiac remodeling post-MI. Methods and Results: To generate mice with Myeloid-specific Plpp3 deletion , female Plpp3 fl/fl mice were crossed to male Plpp3 mice expressing Cre recombinase under the control of the LysM promoter to generate lysm-plpp3. mice. These mice and their littermate control underwent MI or sham surgery. Inflammatory cell content was assessed using flow cytometry and immunohistochemistry. Cardiac function and scar size were assessed by echocardiography and Mason Trichrome staining, respectively. Increased number of Ly6C hi monocytes (CD45 + /Ly6C/G hi /CD115 hi ) and pro-inflammatory macrophages (CD45 + /F4-80 + /CD11b + ) (CD45 + /F4-80 + /CD86 + ), in cardiac tissue of Cre+LysM mice was observed compared to Cre-fl/fl littermate controls during peak post-MI inflammation, as assessed by flow cytometry ( Fig.1A-C ). This increase in inflammatory cells and inflammation may be the consequence of the significant increase in bone marrow and spleen progenitor cell count and proliferation. Moreover, Cre+LysM mice cardiac functional recovery was also reduced significantly ( Fig. 1D-F ) as assessed by echocardiography. Conclusion: Myeloid-specific Plpp3 deletion increases the deleterious effects of inflammation on the ischemic myocardium and ATX/ LPA signaling could represent a novel therapeutic target for future clinical studies of coronary heart diseases.