bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cyclic ADP-Ribose Increases Human Mesenchymal Stem Cells Proliferation through the Modulation of Spontaneous Ca2+ Oscillation.
Authors: Tao Rong , Lau Chu-Pak , Lee Hon-Cheung , Li Gui-Rong 
Published: October 2019

Abstract
Human mesenchymal stem cells (hMSCs) play an important role in the regulation of hematopoietic microenvironment. Cyclic ADP-ribose (cADPR), a universal Ca2+ mobilizer produced by hMSCs in the bone marrow, has been demonstrated to be an important factor in the regulation of hematopoietic stem cells proliferation. However, its biological effects on hMSCs are not understood. The present study was designed to investigate the effects of cADPR on Ca2+ signaling, cell proliferation and differentiation in hMSCs. Ca2+ activity in hMSCs was measured with a confocal microscope. It was found that cADPR increased the frequency of spontaneous Ca2+ oscillatory transient, and the effect was abolished by pretreatment with the specific cADPR antagonist 8-Br-cADPR, but not by ryanodine, though the ryanodine receptors are the classic targets of cADPR in other types of cells. RT-PCR showed no gene expression for ryanodine receptors in hMSCs. Interestingly, the cADPR-induced increase of spontaneous Ca2+ oscillation was abolished by the TRPM2 channel inhibitors econazole and clotrimazole. The downregulation of TRPM2 channels with siRNA technique had no significant effects on spontaneous Ca2+ oscillation; however, it abrogated the cADPR-induced increase of spontaneous Ca2+ oscillation. The effects of cADPR on cell proliferation and differentiation were then investigated in hMSCs. We found that cADPR increased hMSCs proliferation, but had no significant effects on osteogeneic and adipogeneic differentiation. Importantly, the effect of cADPR on hMSCs proliferation was antagonized by 8-Br-cADPR or by the selectively knockdown of TRPM2 channels. In conclusion, our results demonstrate the novel information that cADPR regulate cell proliferation by increasing spontaneous Ca2+ oscillation mediated by TRPM2 channels, but has no effects on osteogeneic and adipogeneic differentiation in hMSCs.
The Biological Characteristics of Osteoblasts Derived from Myelodysplastic Syndrome Patients.
Authors: Chen Wen-ming , Chen Zi-xing , Cen Jian-nong , He Jun , Jiao Xiao-li , Wu Ya-fang , Zhang Jun , Qiu Qiao-cheng , Dai Lan 
Published: October 2019

Abstract
It was hypothesized that osteoblasts play a central role in hematopoiesis, and it has been shown that osteoblasts produce many factors essential for the survival, renewal, and maturation of hematopoietic stem cells (HSCs). By using human fetal osteoblastic cell line hFOB1.19 as a model of control, we investigated the biological characteristics of osteoblasts derived from patients with myelodysplastic syndrome (MDS) and their hematopoietic supportive function in vitro. MSCs isolated from bone marrow of MDS patients and normal donors were cultured and checked for their morphology, immunophenotype, CFU-F forming capacity and the expression of hematopoietic cytokines. A feeder layer was prepared by osteoblasts induced from 3rd generation of cultured MSCs and treated with mitomycin C. Ficoll-isolated bone marrow mononuclear cell from normal donors were then seeded on the feeder layer to co-culture in vitro without exogenous cytokines. FCM revealed that both MSCs and hFOB cells were positive for CD44, CD73(SH3), CD105(SH2) and CD90 (Thy1), but negative for CD34, CD45, HLA-DR. RT-PCR found that hFOB cells expressed mRNA of SCF, IL-6, IL-11, SDF-1, GM-CSF and G-CSF. MSCs obtained from MDS patients and normal donors were displaying fibroblastoid morphology. Their growth pattern, immunophenotype and CFU-F forming capacity were similar (P >0.05). Without exogenous cytokines, the osteoblasts derived from MDS could sustain GM-CFC survival for at least 3 weeks. The CFU-GM yield from cells in upper layer of co-culture was not different from those of control in hematopoiesis supportive experiments in vitro (P>0.05). RT-PCR clearly demonstrated that the cultured BM-MSCs from normal donor expressed mRNA of SCF, SDF-1, IL-6, and IL-11. As the MSCs differentiated toward osteoblasts, the expression of G-CSF could be detected, whereas GM-CSF remained undetectable. The same expression profile of above cytokines were also seen in osteoblasts induced from BM-MSCs of MDS patients. In conclusion, osteoblasts may play a pivotal role in hematopoiesis. The biological characteristics of osteoblasts from bone marrow of MDS patients were generally not different from those of osteoblasts in bone marrow of normal controls. Both of them could support survival of GM-CFC hematopoietic progenitor cells in vitro, according to their expression of multiple cytokines. These findings suggested that the osteoblasts derived from MDS patients may not be involved in the malignant process.
Nitric Oxide from Mesenchymal Stem Cells Regulated T-Cells through Inhibiting Expression of the T-bet.
Authors: Liu Gexiu , He Dongmei , Tan Guangxiao 
Published: October 2019

Abstract
INTRODUCTION: More recently, experimental evidence and preliminary clinical studies have demonstrated that mesenchymal stem cells (MSCs) have an important immune modulatory function. Many reports have shown that MSCs regulate functions of T-cells and suppress its proliferation, but the molecular mechanisms are poorly understood. Here We found that T-bet expression in T cells is suppressed in the presence of MSCs, and nitric oxide (NO) is involved in the regulation of T-cell status and T-bet expression.
METHODS: Adult C57BL/6 mouse bone marrow MSCs were characterized by surface molecules, the typical spindle-shaped morphology, and the ability of differentiation. T-lymphocytes were isolated from the spleen of rats by using mouse CD4, CD8, and CD19 MACS beads, of which the purity determined by flow cytometry was more than 85%. Responding recipient cells (1×10 5) and an equal number of irradiated (3000 cGy) donor stimulating cells or MSCs were cocultured. Cell proliferation was assessed using enzyme-linked immunosorbent assay (ELISA), in accordance with the BrdU user’s manual.
RESULTS: Flow cytometry showed that expression of the activation markers CD25 and CD69 on CD4 or CD8 T cells increased significantly in the presence of stimulating cells, and the production of IFN-and expression of its T-bet in T cells in response to allogeneic stimulation increased to 18 times and 15times, respectively. However, after 24 hours in the presence of MSCs, IFN-γ in T cells was diminished, expression of the T-bet was reduced more than 10 times, T cell proliferation was suppressed by 73–92% in response to allogeneic stimulation or in response to ConA activation. Reverse transcriptase-polymerase chain reaction and Western blot analysis showed that the expression of iNOS in MSCs cocultured with activated splenocytes increased significantly, but not in MSCs alone, or splenocytes alone. The immunofluorescence studies showed that iNOS was exclusively expressed by large adherent CD45− cells, which correspond to MSCs. To further determine a functional immunosuppressive role for NO expressed by MSCs, N-nitro-L-arginine methyl ester, the NO-specific inhibitor was used. As a result, it reversed the suppression of T-cell proliferation and the T-bet expression.
CONCLUSION: Taken into account of T-bet’s functions in T cells, these results suggest that NO produced by MSCs is one of the major mediators of T-cell suppression by MSCs.
Long-Term Reversal of Renal Anemia in Rat by Erythropoietin Gene Modified Mesenchymal Stem Cell Transplantation Beneath the Renal Capsule, Which Is Effectively Regulated by Hypoxia.
Authors: Liu Gexiu , He Dongmei , Tan Guangxiao 
Published: October 2019

Abstract
INTRODUCTION: Patients with chronic renal failure usually require exogenous erythropoietin (EPO) to alleviate anaemia resulting from inadequate epo production by the kidneys. Although this treatment is effective, recombinant EPO could cause hypertension, its antibodies, and other side-effects. EPO gene therapy is more attractive since it could be more economical and more convenient for the long-term management of the disease. As targets for gene therapy, mesenchymal stem cells (MSCs) are more easily expanded, lower immunogenic, and more stable for expressing exogenous genes than other cells. Here we describe an approach of transplantation of the erythropoietin gene modified mesenchymal stem cells beneath the renal capsule to treate Wistar rats with relative erythropoietin deficiency.
METHODS: we constructed an EPO-expressed vector pGenesil-HRE-EPO directed by hypoxia response element (HRE)/SV40 promoter, and transduced it into MSCs prepared by adherently culturing rat bone marrow. The EPO-gene modified MSCs (Epo+) in vitro growed normally and stably secreted high level of functional EPO in hypoxia condition, which produced from 3.23 × 10(5) to 9.98 × 10(5) mIU of EPO per 10(6) cells per 24 hr. To determine the role of these MSCs to treate renal anemia in vivo, EPO+, EPO (vector pGenesil-1 or normal) MSCs were injected into rats with renal anemia beneath the renal capsule, the skin, or in muscles (4 × 10(7) cells/rat). Renal anemia rats had been induced by feeding with adenine, whose hematocrit levels were at 33.5 +/− 4.1%, the plasma Epo concentration was 6.5 ± 1.7 mU/ml. EPO concentration was determined by ELISA. During whole experiment, we measured rat body weight and blood pressure, and obtained blood samples regularly.
RESULTS: 2wk after transplantation, the hematocrit levels in anemia rats injected Epo+ MSCs beneath the renal capsule, the skin, or in muscles markedly increased to 67.2 ± 5.9%, 61.5 ± 5.7%, and 53.8 ± 5.2%, and 10wk after transplantation to 68.0 ± 5.6%, 52.7 ± 5.8%, and 40.6 ± 5.5%, respectively. Moreover, the plasma Epo concentration of anemic rats injected Epo+MSCs beneath the renal capsule, the skin, or in muscles was considerably increased to 158 ± 35 mU/ml, 136 ± 36 mU/ml, and 114 ± 42 mU/ml at 2 wk, whereas that of anemic rats injected EPO MSCs was not increased significantly. All rats injected Epo+MSCs did not have polycythemia or severe hypertension. Survival time of anemic rats injected Epo+MSCs beneath the renal capsule is longest among anemic rats. Histological examination by fluorescent microscopy showed that implanted MSCs beneath the renal capsule survived best.
CONCLUSIONS: EPO gene-modified mesenchymal stem cells can be transplanted in renal anemia rats, and can produce sufficiently active EPO to correct anemia. Transplantation of the cells beneath the renal capsule is best.
Maintenance and Expansion of Hematopoietic Stem/Progenitor Cells in a Biomimetic Hematopoietic Cells Niche.
Authors: Liu Ting , Tan Jing , Hou Li , Meng Wentong , Wang Yuchun 
Published: October 2019

Abstract
As main hematopoietic organ, bone marrow have three dimensional microenvironment for hematopoietic stem/progenitor cells grow in, so call “hematopoietic cells niche”, which are composed by stromal cells and extracelluar matrix. The interactions of cell to cell and cell to matrix between stem/progenitor cells with hematopoietic niche are facilitated by its three dimensional conformation. The biology behaviors of hematopoietic stem cells are mediate by many signal transductions between stem/progenitor cells with their corresponding microenvironment. Now there are strong evidence from animal model study suggests that osteoblasts might play an essential role in creation of a hematopoietic stem cell niche and thereby regulation of stem cell maintenance, proliferation, and maturation. In light of the structure-function relationship of bone marrow topography, we conceived a biomimetic culture system (3D culture system) with bio-derived bone as framework, composited with human marrow mesenchymal stem cells, and induced the cells into osteoblasts to simulate the effects of hematopoietic osteobalst niche. CD 34+ cells or mononuclear cells separated from umbilical cord blood were cultured for 2∼5 weeks in the 3D culture system and also in a conventional 2D culture system as control without additional cytokine supplement. Based on our results, higher expression of extracelluar matrix and N-cadherin were observed in 3D culture system compared to 2D system. At 2 weeks culture, 3D culture system showed higher number of CD34+ cells and CD34+/CD38- cells when compared with the input (P<0.05), the increased cells were predominant CD34+/CD38-cells. Although CD34+ cells were decreased at 5 weeks culture; nevertheless, CD34+/CD38- cells were still maintained at high level. We also observed that imbedding MNCs with a higher percentage of CD34+/CD38-cells cultured in 3D system (P<0.05), which may represent a down regulation of CD38 phenotype during culture. The function of cultured cells was evaluated in colony forming unit (CFU) assay and long term culture initial cell (LTC-IC) assay. 3D system produced higher expansion of CFU progenitors than 2D system (7.13–8.89 times vs. 1.22–1.31times) after 2 weeks culture. Of note, the colony distribution of 3D system manifested higher percentage of BFU-E and CFU-GEMM, while 2D system showed mainly CFU-GM. LTC-IC represents the primitive progenitor, 3D system showed a 6.2 times increment over input after 2 weeks culture. Furthermore, it was competent to maintain the immaturation of hematopoietic progenitor cells (HPCs) over 5 weeks. This study demonstrated that our 3D culture system constructed with the bio-derived bone composited with induced osteoblast is capable to allow maintenance and expansion of primitive hematopoietic progenitor cells in vitro. It may open a new avenue to study HSCs/HPCs behaviors and to achieve sustained primitive progenitor cell expansion.
Transvection Mediated by the Translocated Cyclin D1 Gene in Mantle Cell Lymphoma.
Authors: Epner Elliot M. , Liu Hui , Wang Jing , Thayer Mathew 
Published: October 2019

Abstract
Cyclin D1 expression in B cells is deregulated by chromosome translocations involving the immunoglobulin heavy chain (IgH) locus in mantle cell lymphoma (MCL). Gene targeting experiments produced MCL cell lines that had lost the translocated t(11;14) and no longer expressed cyclin D1. In these cyclin D1 (−) cells, the nonrearranged cyclin D1 (CCND1) locus reverts from CpG hypomethylated to hypermethylated. Reintroduction of the translocated chromosome by somatic cell hybrid fusion induces loss of methylation at the unrearranged CCND1 locus. Thus, the translocated chromosome exerts a transallelic effect on the unrearranged CCND1 locus in B cells that resembles transvection in Drosophilia. Control somatic cell fusion experiments with a nontranslocated cyclin D1 locus do not demonstrate transvection effects. We also have evidence for pairing of the translocated and nontranslocated cyclin D1 loci in MCL cell lines and MCL patient samples. This pairing is not related to DNA replication, as it is observed in flow sorted G1 cells and not in cyclin D1 expressing breast cancer cells or in B lymphocytes. In addition, to pairing of the cyclin D1 loci, we also have demonstrated translocation specific small RNAs in MCL cells upstream of the cyclin D1 gene.
Involvement of STAT3 Transcription Factor in Disseminated Nasal-Type Natural Killer Cell Lymphomas.
Authors: Coppo Paul , Gouilleux-Gruart Valérie , Bouchet Sandrine , Huang Yenlin , Dartigues Peggy , Gaulard Philippe , Agbalika Felix , Douay Luc , Lassoued Kaiss , Gorin Norbert C. 
Published: October 2019

Abstract
Disseminated nasal-type natural killer (NK) cell lymphoma is an aggressive disease with very poor prognosis. The usual chemoresistance of this disease led us to explore the possible role of the transcription factor STAT3 in oncogenesis. For this, we established and characterized a continuous interleukin (IL)2-dependent NK cell line (MEC04) from a patient with a fatal nasal-type NK cell lymphoma. MEC04 cells phenotype was CD3−, CD7+, TCR−, CD16−, CD56+bright, p58−, p70−, NKP−, and NKG2A+, they harbored poor cytotoxic activity against K562 cells, and spontaneously secreted interferon-γ, IL-10 and vascular-endothelium growth factor in vitro, suggesting that malignant cells arise from the CD56+bright regulatory NK cell population. Injection of MEC04 cells to NOD/SCID mice led to a fatal multiorgan infiltration by malignant cells. We show by immunohistochemistry that STAT3 is phosphorylated in Y705 dimerization residue in MEC04 and YT cell lines, and on biopsies from 6/6 patients with nasal-type NK cell lymphoma, suggesting that STAT3 may be oncogenic in this disease. By contrast, Y705 residue was not phosphorylated on 2 patients with hepatosplenic γδT-cell lymphoma. By confocal microscopy, we showed that STAT3 was located in nucleus in MEC04 cells. Y705 STAT3 phosphorylation involved JAK2, since exposure of cells to AG490 inhibitor inhibited Y705 STAT3 phosphorylation, and correlated in a dose-dependent growth inhibition at 24 hours and 48 hours. By using transducible TAT-STAT3b or TAT-STAT3Y705F recombinant proteins (a dominant-negative form of STAT3 [STAT3b isoform], and a STAT3 protein mutated on Y705 residue which prevents STAT3 dimerization [STAT3Y705F]), we were able to demonstrate that inhibition of STAT3 activation increased mortality and decreased proliferation by more than 50 p. cent at 24 hours and 48 hours, which correlated with decreased expression of 2 STAT3 target genes (Bcl-XL and c-Myc). These results suggest that STAT3 protein has a major role in the oncogenic process of nasal-type NK cell lymphomas, and may thus represent a promising therapeutical target.
Synergistic Effect of Magnetic Nanopartical Fe304, Au and Daunomycin on K562/A02.
Authors: Chen Bao-An , Dai Yong-Yuan , Wang Xue Mei , Zhang Ren-Yun , Gao Feng , Chen Jian , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Ding Jia-Hua , Chen Ning-Na , Pei Xiao-Ping 
Published: October 2019

Objective: This paper was to study the reversal effect of magnetic Nano-Fe3O4 or Nano-Au with DNR, on multidrug resistance cell line K562/A02 and to investigate the reversal mechanism of this combination, and to provide theoretic evidence for the clinical application of them as resistance modifying agents.
Method: The IC50 (the concentration causing 50% inhibition of cell growth) of DNR, Nano-Fe3O4 and Nano-Au respectively were assayed by MTT method.The drug-loaded nanoparticles were prepared by solvent diffusion method.Some nanoparticales, volume ratio from 1.5% to 50%, were combined with some DNR to find the best combination to prepare best drug-loaded nanopartilces.At last, the K562/A02 cells was treated with the composite of 25% nanoparticales and 10mg/L DNR, which MDR1 mRNA was assayed by RT-PCR;intracellular drug concentration and the apoptosis was determined by fluorometry and confocal fluorescence microscope.
Results: The IC50 of DNR for K562/A02 and K562 cells were 23.23mg/L and0.307mg/L respectively.Two nanoparticles themselves have not evident cytotoxic effect to K562/A02 and K562 cells.Pretreating K562/A02 cells with the composite of 25% nanoparticales and 10mg/L DNR for 48 hours partially restored the sensitivity of K562/A02 cells to DNR;K562/A02 showed apoptotic characteristics after treated with this composite;drug-loaded nanopartilces elevated the intracellular DNR accumulation in K562/A02 and its MDR1 mRNA were down regulated.Data was analyzed by SPSS 11.5 software and expressed as mean ± SD.
Conclusions:
Nano- Fe3O4 or Nano-Au can increases the intracellular free DNR concentration of the K562/A02 cells, which lead to more K562/A02 cells apoptosis. Two nanoparticles themselves could not lower the MDR1 gene expression of the K562/A02 cells, but they degraded the MDR1 gene level with combine DNR.
These results suggested that Nano- Fe3O4 or Nano-Au with DNR can reverse the resistance of K562/A02 cells significantly.
Research of Synergetic Reversal in Adriamycin - Resistance by the Application of Magnetic Nanoparticle of Fe3O4 and Tetrandrine in K562/A02 Leukemic Cells.
Authors: Chen Bao-An , Shun Qian , Wang Xue-Mei , Zhang Ren-Yu , Gao Feng , Chen Jian , Gao Chong , Wang Jun , Ding Jia-Hua , Zhao Gang , Chen Ning-Na , Pei Xiao-Ping , Dai Yong-Yuan , Shun Ning 
Published: October 2019

Objective: To prepare functionalized Fe3O4-magnetic nanoparticles(Fe3O4-MNPs) loaded with adriamycin(ADM) or Fe3O4-MNPs co-polymerized with ADM and tetrandrine(Tet) to investigate whether the temperature, time or ratio of drug to nanoparticles influences the polymerization. To study the reversal role that the drug-loaded nano-composites play in K562 and one of its resistant cell line K562/A02 and to learn the reversal mechanism of this kind of treatments so as to provide theoretic evidence for the clinical application of them as resistance modifying agents.
Method: The drug-loaded nanoparticles were prepared using mechanical absorption polymerization process in different condition of 4° or 37° for 24h or 48h. To investigate whether Fe3O4-MNPs loaded with ADM and Tet would play a synergetic reverse role in multidrug resistant cell, the drug-loaded nanoparticles by mechanical absorption polymerization were prepared to act with K562 and one of its resistant cell line K562/A02. The survival of cells which were cultured with drug-loaded nano-composites for 48h was observed through MTT assay, the growth inhibition efficacy of cells was calculated then. Using cells under the same condition described before, we took use of fluorescence microscope to measure fluorescence intensity of intracellular ADM at a wavelength of 488nm. P-glycoprotein (P-gp) was analyzed with flow cytometer. The expression of mdr1 mRNA was measured by RT-PCR.
Results: The results showed that the growth inhibition efficacy of both the two cells increased with augmenting concentration of Fe3O4-MNPs which were loaded with drugs. The condition of 4° and 48h was significantly better than that of 37° and 24h respectively. Both Fe3O4-MNPs loaded with ADM or Fe3O4-MNPs co-polymerized with ADM and Tet elevated the intracellular ADM accumulation in K562/A02. However, no linear correlation was found between fluorescence intensity of intracellular adriamycin and augmenting concentration of Fe3O4-MNPs in K562/A02. Tet could downregulate the level of mdr-1 gene and decrease the expression of P-gp. Furthermore, Tet polymerized with Fe3O4-MNPs reinforced this downregulation, causing a 100-fold more decrease in mdr1 mRNA level, but did not reduce total P-gp content.
Conclusions: Fe3O4-MNPs can load ADM or both ADM and Tet by mechanical absorption polymerization, which depends on proper temperature and time. Furthermore, the drug-loaded nano-composites have the ability in multidrug resistance reversal. Fe3O4-MNPs loaded with ADM or Tet can enhance the effective accumulation of the drugs in K562/A02 and Fe3O4-MNPs loaded with Tet obviously reverse multidrug resistance by reinforcing mdr1 gene downregulation. Functionalized Fe3O4-MNPs loaded with Tet probably have synergetic effect on reversal in multidrug resistance.