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Panax Notoginseng Saponins Induced Up-Regulation,Phosphorylation and Binding Activity of MEK,ERK,AKT,PI-3K Protein Kinases and GATA Transcription Factors in hematopoietic cells

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Chinese Journal of Integrative Medicine 2013年第2期19卷 112-118页

作者：孙馨高瑞兰林筱洁徐卫红陈小红 Department of Oncology Zhejiang Provincial People's Hospital Institution of Hematology Research The First Affiliated HospitalZhejiang Chinese Medical University

Objective： To investigate the effects of panax notoginseng saponins （PNS） on expression, regulation and phosphorylation of multiple protein kinases in mitogen activated protein kinase （MAPK） intracellular signal pathway and GATA transcription factors in hematopoietic cells, so as to explore its mechanism of proliferation and differentiation activity on hematopoiesis. Methods： The human granulocytic HL-60, erythrocytic K562, megakaryocytic CHRF-288 and Meg-01 cell lines were treated by PNS, the positive control of K562, CHRF-288 ceils treated by recombination human erythropoietin （Epo） and thrombopoietin （Tpo） respectively. The total cell lysate and nuclei protein were extracted after being treated by PNS, subsequently, analyzed by both Western blot and immune-precipitation. Meanwhile, the nuclei extract was performed for electrophoretic mobility shift assay （EMSA） by using 32p radio labeled double-stranded GATA consensus oligonucleotide. Results： The expression levels of kinase MEK-1, MEK-2, ERK-1, ERK-2, AKT-1, AKT-2 and PI- 3K were increased by PNS treatment to different extent in four cell lines, depending on cellular heterogeneity and sensitivity to PNS, also phosphorylation of MEK-1, ERK-1 was differentially promoted by PNS respectively （P〈0.05, 0.01, 0.001）. The expression levels of transcription factors GATA-1 and GATA-2 were increased, moreover, their DNA binding activities were raised dramatically in PNS treated K562, CHRF-288 and Meg-01 cells compared with the controls respectively （P〈0.05, 0.01, 0.001）. The positive control of K562, CHRF-288 cells treated by Epo or Tpo respectively also displayed up-regulation of protein kinases and GATA transcription factors respectively （P〈0.05, 0.01, 0.001）. Conclusion： The results indicated that intracellular signal pathway initiated by PNS was involved in MAPK pathway and transcription factors of GATA family in hematopoietic cells. PNS displayed the role to promote proliferation and differentiation, by means of incre

关键词： panax notoginseng saponins hematopoietic cells protein kinase phosphorylation transcription factor

Experiments on Gene Transferring to Primary hematopoietic cells by Liposome

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2000年第4期20卷 287-289页

作者：胡丽华张波 Department of Blood Transfusion Xiehe Hospital Tongji Medical University Wuhan

Summary Liposomes have showed many advantages in mediating exogenous gene into many cell types in vitro and in vivo. But few data are available concerning gene transfer into hematopoietic cells. In this report, we described two-marker genes (Neo R and Lac Z) co-transferred into hematopoietic cells of human and mouse by using liposome in vitro. The efficiency of gene transfer was tested by X- gal staining and observation of colony formation. The X-gal blue staining rate of transduced cells was about (13. 33±2.68)% in human and about (16. 28±2. 95) % in mouse without G418 selection. After G418 selection, the blue cell rate was (46. 06±3. 47)% in human and (43. 45±4. 1)% in mouse, which were markedly higher than those before selection, suggesting that high-efficiency gene transfer and expression could be attained in primary hematopoietic cells using this easy and harmless transduction protocol. At the same time, this protocol provided experimental data for clinicians to in- vestigate the biology of marrow reconstitution and trace the origin of relapse after autologous bone marrow transplantation for the patients with leukemia.

关键词： liposome gene transfer hematopoietic cells

Adoptive transfer of Pfkfb3-disrupted hematopoietic cells to wild-type mice exacerbates diet-induced hepatic steatosis and inflammation

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Liver Research 2020年第3期4卷 136-144页

作者： Xin Guo Bilian Zhu Hang Xu Honggui Li Boxiong Jiang Yina Wang Benrong Zheng Shannon Glaser Gianfranco Alpini Chaodong Wu Department of Nutrition Texas A&M UniversityCollege StationTXUSA Department of Nutrition and Food Hygiene School of Public HealthCheeloo College of MedicineShandong UniversityJinanChina Department of VIP Medical Service Center The Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina Medical Physiology Texas A&M University College of MedicineBryanTXUSA Hepatology and Gastroenterology MedicineIndiana UniversityIndianapolisINUSA Richard L.Roudebush VA Medical Center IndianapolisINUSA

Background and objectives:Hepatic steatosis and inflammation are key characteristics of non-alcoholic fatty liver disease(NAFLD).However,whether and how hepatic steatosis and liver inflammation are differentially regulated remains to be *** that disruption of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(Pfkfb3/iPfk2)dissociates fat deposition and inflammation,the present study examined a role for Pfkfb3/iPfk2 in hematopoietic cells in regulating hepatic steatosis and inflammation in ***:Pfkfb3-disrupted(Pfkfb3^(+/-))mice and wild-type(WT)littermates were fed a high-fat diet(HFD)and examined for NAFLD ***,bone marrow cells isolated from Pfkfb3^(+/-)mice andWT mice were differentiated into macrophages for analysis of macrophage activation status and for bone marrow transplantation(BMT)to generate chimeric(WT/BMT-Pfkfb3^(+/-))mice in which Pfkfb3 was disrupted only in hematopoietic cells and control chimeric(WT/BMT-WT)*** latter were also fed an HFD and examined for NAFLD *** vitro,hepatocytes were co-cultured with bone marrowderived macrophages and examined for hepatocyte fat deposition and proinflammatory ***:After the feeding period,HFD-fed Pfkfb3^(+/-)mice displayed increased severity of liver inflammation in the absence of hepatic steatosis compared with HFD-fed WT *** inflammatory activation was analyzed,Pfkfb3^(+/-)macrophages revealed increased proinflammatory activation and decreased anti-proinflammatory *** NAFLD phenotype was analyzed in the chimeric mice,WT/BMT-Pfkfb3^(+/-) mice displayed increases in the severity of HFD-induced hepatic steatosis and inflammation compared with WT/BMT-WT *** the cellular level,hepatocytes co-cultured with Pfkfb3^(+/-) macrophages revealed increased fat deposition and proinflammatory responses compared with hepatocytes co-cultured with WT ***:Pfkfb3 disruption only in hematopoietic cells exacerbates HFD-induce

关键词： 6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(Pfkfb3/iPfk2) hematopoietic cells Hepatic steatosis Inflammation Macrophages

Single-cell transcriptomic landscape of human blood cells

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National Science Review 2021年第3期8卷 42-52页

作者： Xiaowei Xie Mengyao Liu Yawen Zhang Bingrui Wang Caiying Zhu Chenchen Wang Qing Li Yingying Huo Jiaojiao Guo Changlu Xu Linping Hu Aiming Pang Shihui Ma Lina Wang Wenbin Cao Shulian Chen Qiuling Li Sudong Zhang Xueying Zhao Wen Zhou Hongbo Luo Guoguang Zheng Erlie Jiang Sizhou Feng Lixiang Chen Lihong Shi Hui Cheng Sha Hao Ping Zhu Tao Cheng State Key Laboratory of Experimental Hematology and National Clinical Research Center for Blood Diseases Institute of Hematology and Blood Diseases HospitalChinese Academy of Medical Sciences &Peking Union Medical College Center for Stem Cell Medicine and Department of Stem Cell & Regenerative Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Cancer Research Institute School of Basic Medical Science Central South University School of Life Sciences Zhengzhou University

High throughput single-cell RNA-seq has been successfully implemented to dissect the cellular and molecular features underlying ***,an elaborate and comprehensive transcriptome reference of the whole blood system is ***,we profiled the transcriptomes of 7551 human blood cells representing 32 immunophenotypic cell types,including hematopoietic stem cells,progenitors and mature blood cells derived from 21 healthy *** high sequencing depth and coverage,we constructed a single-cell transcriptional atlas of blood cells(ABC) on the basis of both protein-coding genes and long noncoding RNAs(lncRNAs),and showed a high consistence between ***,putative lncRNAs and transcription factors regulating hematopoietic cell differentiation were *** common transcription factor regulatory networks were activated in neutrophils and monocytes,lymphoid cells dramatically changed their regulatory networks during ***,we showed a subset of nucleated erythrocytes actively expressing immune signals,suggesting the existence of erythroid precursors with immune ***,a web portal offering transcriptome browsing and blood cell type prediction has been ***,our work provides a transcriptional map of human blood cells at single-cell resolution,thereby offering a comprehensive reference for the exploration of physiological and pathological hematopoiesis.

关键词： human hematopoietic cells lncRNAs transcription factor single-cell RNA-seq

Cellular Reprogramming of Human Peripheral Blood cells

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Genomics, Proteomics & Bioinformatics 2013年第5期11卷 264-274页

作者： Xiao-Bing Zhang Department of Medicine Loma Linda University

Breakthroughs in cell fate conversion have made it possible to generate large quantities of patient-specific cells for regenerative medicine. Due to multiple advantages of peripheral blood cells over fibroblasts from skin biopsy, the use of blood mononuclear cells （MNCs） instead of skin fibroblasts will expedite reprogramming research and broaden the application of reprogramming technology. This review discusses current progress and challenges of generating induced pluripotent stem cells （iPSCs） from peripheral blood MNCs and of in vitro and in vivo conversion of blood cells into cells of therapeutic value, such as mesenchymal stem cells, neural cells and hepatocytes. An optimized design of lentiviral vectors is necessary to achieve high reprogramming efficiency of peripheral blood cells. More recently, non-integrating vectors such as Sendai virus and episomal vectors have been successfully employed in generating integration-free iPSCs and somatic stem cells.

关键词： Reprogramming Peripheral blood hematopoietic cells Induced pluripotent stemcells Cell fate conversion

Gene Expression Profiling of Human c-Kit Mutant D816V

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Journal of Cancer Therapy 2016年第6期7卷 439-454页

作者： Shilpa Sharma Gurudutta Gangenahalli Division of Stem Cell & Gene Therapy Research Institute of Nuclear Medicine & Allied Sciences (INMAS) New Delhi India

The tyrosine kinase receptor III, c-Kit/stem cell factor receptor and its ligand, human stem cell factor (huSCF) are the predominant regulator of mitogenesis in the hematopoietic stem and progenitor cells. However, gain-of-function mutations alter c-Kit auto-regulatory mechanisms to aberrant c-Kit signaling, leading to the onset or progression of cancerous transformations. The most common mutation of c-Kit is the substitution of aspartic acid residue in position 816 to valine (D816V), which is majorly responsible for its ligand-independent constitutive activation, and is implicated in hematopoietic malignancies. Currently, molecular targeted therapy is increasingly becoming a hot spot due to its specificity and low toxicity. As the molecular mechanisms responsible for D816V-c-Kit mediated tumorogenicity are largely unknown, in this study, we aimed to investigate the D816V-c-Kit signaling mediated downstream molecular targets. Specifically, we created c-Kit active mutant form D816V and performed inducible gene expression of mutant D816V-c-Kit in monomyelocytic cell line U937. Mutant D816V-c-Kit expressing cells revealed significantly enhanced cellular mitogenic activity compared to wild-type c-Kit expressing cells independent of huSCF. To examine the molecular targets regulating tumorogenic proliferation, we evaluated the consequences of mutant D816V-c-Kit expression on downstream gene expression profile by high throughput microarray technology. The levels of some of the relevant genes (PIK3CB, eIF4B, PRKCDBP, MOAP1) were validated by quantitative polymerase chain reaction. SLA, STAT5B, MAP3K2 and MAPK14 emerged as important downstream molecular targets of mutant D816V-c-Kit. Further, by dissecting the signaling pathways, we also demonstrated that the D816V-c-Kit mediated hematopoietic cell proliferation is dependent on molecular target p38 MAP kinase.

关键词： c-Kit Mutant hematopoietic cells Microarray Gene Expression Proliferation

The protective effect of new aryl sulfone derivatives against radiationinduced hematopoietic injury

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The protective effect of new aryl sulfone derivatives agains...