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Zhou H.-Y.,Chinese Academy of Sciences | Meng Z.-Y.,Beijing Institution of Transfusion Medicine | Dou G.-F.,Beijing Institution of Transfusion Medicine | Ma J.-L.,Chinese Academy of Sciences | And 2 more authors.
Yaoxue Xuebao | Year: 2010

This study is to elucidate the metabolic pathway of 1, 2-[bis (1, 2-benzisoselenazolone-3 (2H)-ketone)]-ethane (BBSKE) in rats. Rats were administrated with a single dose of BBSKE 200 mg·kg-1. The metabolites in rat urine, feces, bile and plasma were identified by LC-MS n analysis. The characterization of fragment ions from LC-MS n chromatography and mass spectrometry was applied to the investigation of structures of metabolites. Three phase I metabolites were detected in rat urine and feces. Two of them were also found in plasma and one existed in bile. These products were derived from oxidized, methylated and S-methylated BBSKE, separately. One phase II glucuronide of BBSKE was also found in bile. Therefore, it is possible that BBSKE was metabolized by oxidization, methylation and glucuronidation.


Liu Y.-X.,Chinese PLA General Hospital | Dong X.,Chongqing Medical University | Dong X.,General Hospital of Beijing Military Command | Gong F.,Beijing Institution of Transfusion Medicine | And 7 more authors.
PLoS ONE | Year: 2015

Suppressor of cytokine signaling 3 (SOCS3) plays an important role in mice fetal liver erythropoiesis, but the roles of SOCS3 in human hematopoietic stem cells (HSCs) have not been well investigated. In the present study, lentiviral small interference RNA expression vectors (shRNA) of SOCS3 were constructed and stably transferred into HSCs. We found that SOCS3 knockdown induced erythroid expansion in HSCs. Conversely, Ectopic expression of SOCS3 in progenitor cells blocked erythroid expansion and erythroid colony formation of HSCs. To further explore the involved mechanism, we compared gene expression profiles of SOCS3-shRNA tranduced HSCs with that of control HSCs by whole genome microarrays. The results indicated that cell developmental process related genes, especially hematopoietic lineage-specific genes, associated with the responses to SOCS3 in HSCs. Downexpression of SOCS3 in HSCs or differentiated erythroid progenitor cells induced a transcriptional program enriched for erythroid development relative genes. Our results proved that SOCS3 down-expression induced lineage commitment towards erythroid progenitor cell fate by activation of erythroid-specific gene in HSCs and provided new insight into the mechanism of erythropoietic development. © 2015 Liu et al.


PubMed | Beijing Institution of Transfusion Medicine, Chinese PLA General Hospital and Chongqing Medical University
Type: Journal Article | Journal: PloS one | Year: 2015

Suppressor of cytokine signaling 3 (SOCS3) plays an important role in mice fetal liver erythropoiesis, but the roles of SOCS3 in human hematopoietic stem cells (HSCs) have not been well investigated. In the present study, lentiviral small interference RNA expression vectors (shRNA) of SOCS3 were constructed and stably transferred into HSCs. We found that SOCS3 knockdown induced erythroid expansion in HSCs. Conversely, Ectopic expression of SOCS3 in progenitor cells blocked erythroid expansion and erythroid colony formation of HSCs. To further explore the involved mechanism, we compared gene expression profiles of SOCS3-shRNA tranduced HSCs with that of control HSCs by whole genome microarrays. The results indicated that cell developmental process related genes, especially hematopoietic lineage-specific genes, associated with the responses to SOCS3 in HSCs.Downexpression of SOCS3 in HSCs or differentiated erythroid progenitor cells induced a transcriptional program enriched for erythroid development relative genes. Our results proved that SOCS3 down-expression induced lineage commitment towards erythroid progenitor cell fate by activation of erythroid-specific gene in HSCs and provided new insight into the mechanism of erythropoietic development.


Yang C.,Beijing Institution of Transfusion Medicine | Ji L.,Beijing Institution of Transfusion Medicine | Ji L.,CAS Institute of Zoology | Shi S.S.,Beijing Institution of Transfusion Medicine | And 4 more authors.
Progress in Biochemistry and Biophysics | Year: 2010

Erythropoietin (EPO) plays an important role in modulating proliferation and differentiation of erythrocytes. The fetal liver stromal cell lines(FLSCs) expressing EPO has been established steadily by lentiviral system. The EPO gene was cloned from human fetal liver by RT-PCR. The EPO recombinant lentiviral plasmid was steadily transfected into FLSCs. The efficiency of virus transfection was identified by expression of enhanced green fluorescence protein (eGFP) analyzed by fluorescence microscope, then the high eGFP espression FLSCs were sorted by fluorescence-activated cell sorting (FACS) according to strong eGFP expression. Analysis of strong eGFP expression was detected by RT-PCR and ELISA. The EPO expression at mRNA level of strong eGFP expression FLSCs are 5.63 and 5.71-fold for the FLSCs no transfected and the FLSCs transfected by the control lentivirus. And at protein level, the content of EPO expression is 263 U/L. Then the supernatant from the EPO transfected FLSCs could induce the CD34+ cell differentiated into hematopoietic cell, especially erythrocytes. This would provide an alternative for cell therapy and blood cell transfusion.


Liu Y.-X.,Beijing Institution of Transfusion Medicine | Liu Y.-X.,First Affiliated Hospital of PLA General Hospital | Yue W.,Beijing Institution of Transfusion Medicine | Ji L.,Beijing Institution of Transfusion Medicine | And 2 more authors.
BMC Developmental Biology | Year: 2010

Background. We recently developed a new method to induce human stem cells (hESCs) differentiation into hematopoietic progenitors by cell extract treatment. Here, we report an efficient strategy to generate erythroid progenitors from hESCs using cell extract from human fetal liver tissue (hFLT) with cytokines. Human embryoid bodies (hEBs) obtained of human H1 hESCs were treated with cell extract from hFLT and co-cultured with human fetal liver stromal cells (hFLSCs) feeder to induce hematopoietic cells. After the 11 days of treatment, hEBs were isolated and transplanted into liquid medium with hematopoietic cytokines for erythroid differentiation. Characteristics of the erythroid cells were analyzed by flow cytometry, Wright-Giemsa staining, real-time RT-PCR and related functional assays. Results. The erythroid cells produced from hEBs could differentiate into enucleated cells and expressed globins in a time-dependent manner. They expressed not only embryonic globins but also the adult-globin with the maturation of the erythroid cells. In addition, our data showed that the hEBs-derived erythroid cells were able to act as oxygen carriers, indicating that hESCs could generate functional mature erythroid cells. Conclusion. Cell extract exposure with the addition of cytokines resulted in robust erythroid -like differentiation of hEBs and these hEBs-derived erythroid cells possessed functions similar to mature red blood cells. © 2010 Liu et al; licensee BioMed Central Ltd.


PubMed | Beijing Institution of Transfusion Medicine
Type: | Journal: BMC developmental biology | Year: 2010

We recently developed a new method to induce human stem cells (hESCs) differentiation into hematopoietic progenitors by cell extract treatment. Here, we report an efficient strategy to generate erythroid progenitors from hESCs using cell extract from human fetal liver tissue (hFLT) with cytokines. Human embryoid bodies (hEBs) obtained of human H1 hESCs were treated with cell extract from hFLT and co-cultured with human fetal liver stromal cells (hFLSCs) feeder to induce hematopoietic cells. After the 11 days of treatment, hEBs were isolated and transplanted into liquid medium with hematopoietic cytokines for erythroid differentiation. Characteristics of the erythroid cells were analyzed by flow cytometry, Wright-Giemsa staining, real-time RT-PCR and related functional assays.The erythroid cells produced from hEBs could differentiate into enucleated cells and expressed globins in a time-dependent manner. They expressed not only embryonic globins but also the adult-globin with the maturation of the erythroid cells. In addition, our data showed that the hEBs-derived erythroid cells were able to act as oxygen carriers, indicating that hESCs could generate functional mature erythroid cells.Cell extract exposure with the addition of cytokines resulted in robust erythroid -like differentiation of hEBs and these hEBs-derived erythroid cells possessed functions similar to mature red blood cells.

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