OBJECTIVETo investigate the clinicopathological diagnosis and expressions of CD117, CD34, SMA, S-100 protein, Vimentin(Vim) and desmin in gastrointestinal stromal tumors (GISTs).

METHODSA retrospective analysis of the clinical data and the results of various examinations was conducted among 35 patients with pathologically confirmed GISTs undergoing surgical resection. The expressions of CD117, CD34, SMA, S-100, Vim and desmin in the tumor tissues were detected by immunohistochemistry with SP method.

RESULTSIn these GIST cases, the tumors were located mostly in the stomach (n=11), small intestines (n=11), and abdominal cavity (n=5). The main clinical manifestations included abdominal distension, abdominal pain, gastrointestinal bleeding, and abdominal masses. The positivity rates of CD117 and CD34 in the tumors were 94.3% and 91.4%, respectively, both significantly higher than those of SMA, S-100, Vim and Desmin (P<0.001), and also higher than that in leiomyoma (P<0.0001). The positivity rate of Desmin was only 2.9% in the tumors, significantly lower than those of CD117 and CD34 (P<0.05) and that in liomyoma (P<0.001).

CONCLUSIONSGISTs occur mostly in the stomach and small intestines, and endoscopy, ultrasound endoscope and CT examination are effective modalities for diagnosis of GISTs. A definite diagnosis of GISTs can be established in the presence of positive expression of CD117 and CD34 and negative expression of Desmin in the tumor.

Adult ; Aged ; Antigens, CD34 ; biosynthesis ; Biomarkers, Tumor ; biosynthesis ; Desmin ; biosynthesis ; Female ; Gastrointestinal Stromal Tumors ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Proto-Oncogene Proteins c-kit ; biosynthesis ; Retrospective Studies ; S100 Proteins ; biosynthesis ; Sensitivity and Specificity ; Young Adult

Primary leiomyosarcoma of the nipple-areola complex is extremely rare. Less than ten such cases have been reported in English literature so far. Herein we describe a 52-year-old female presenting with a 1.5 cmx1.1 cmx0.7 cm nodular lesion over her left nipple, and leiomyosarcoma was proved by pathological examination of the excised specimen. Positron emitted tomogram (PET) revealed no abnormal signal other than the primary site. Microscopically, this poorly circumscribed tumor was composed of interlacing bundles of smooth muscle cells with bizarre and pleomorphic nuclei, as well as prominent nucleoli. Its mitotic count was up to 7 mitoses per 10 high power fields (HPF). Immunohistochemical study of tumor cells revealed positive stain for alpha-smooth muscle actin and vimentin; and negative for cytokeratin, CD34 and S-100. Left simple mastectomy was undertaken and no residual mass lesion was noted on the resected specimen. Related literatures about the diagnosis and treatment for breast leiomyosarcoma will be presented here.
Antigens, CD34 ; biosynthesis ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; pathology ; Cell Nucleus ; metabolism ; Female ; Humans ; Immunohistochemistry ; methods ; Leiomyosarcoma ; diagnosis ; pathology ; Mastectomy ; Middle Aged ; Myocytes, Smooth Muscle ; pathology ; Nipples ; pathology ; S100 Proteins ; biosynthesis ; Smooth Muscle Tumor ; diagnosis ; pathology ; Treatment Outcome

OBJECTIVETo investigate the potential effect of homologous bone marrow mesenchymal stem cells (MSCs) on repairing peri-tubular capillary cluster (PTCC), and on improving renal tubular and mesenchymal hypoxia condition.

METHODSMonocyte was purified from bone marrow, amplified and identified as MSCs in vitro. Thirty female Wistar rats were randomly divided into 3 groups, the normal control group (Group A), MSCs transplanted group (Group B) and un-transplanted group (Group C). Rats in Group A was administered with drinking water by gastrogavage for 12 weeks, while those in Group B and C were administered with Aristolochia Decoction for 12 weeks to establish chronic aristolochic acid nephropathy (CAAN) model. At the end of the 12th week, 1 ml of MSCs was injected through caudal vein to the rats in Group B, while to those in Group A and C normal saline was injected instead. Blood, urine and kidney tissue of rats were collected at the end of the 16th week for examination, and their kidney tissue were made into serial section for determining the distribution of Y chromosome and CD34 double positive cells, and the pathological, immunohistochemical changes were observed using Western blotting and RT-PCR, etc.

RESULTSY chromosome and CD34 double positive cells could be seen in MSCs transplanted renal tissue in group B. At the end of the 16th week, the PTCC density in Group C and B was (26.47 +/- 1.56)/ 0.13 mm2 and (5.26 +/- 0.78)/0.13 mm2 respectively, and the IOD value of hypoxia inducible factor-1alpha (HIF-1alpha) in them was (6.74 +/- 0.67) x 10(3) and (25 27 +/- 1.46) x 10(3) respectively, all showing significant difference between the two groups (P < 0.01). The content of CD34 was higher in Group B than that in Group C (P < 0.01).

CONCLUSIONHomologous MSCs can enhance the vascular endothelial cells differentiation to repair the PTCC, thus to improve the renal tubular and mesenchymal hypoxia status.

Animals ; Antigens, CD34 ; biosynthesis ; genetics ; Aristolochic Acids ; Blotting, Western ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Capillaries ; abnormalities ; metabolism ; Endothelial Cells ; metabolism ; pathology ; Female ; Immunohistochemistry ; Kidney Diseases ; chemically induced ; pathology ; surgery ; Kidney Tubules ; blood supply ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous

OBJECTIVETo investigate the effect of Xuefu Zhuyu Decoction (XFZYD) on the number, phenotype, cell cycle and colony formation of bone marrow hematopoietic stem cells (HSC) in mice.

METHODSKunming mice were randomly divided into 4 groups: the control group, the low- (3.25 g/kg), middle- (6.5 g/kg) and high-dose (13.0 g/kg) XFZYD groups. After they were medicated by gastrogavage respectively with saline or corresponding dose of XFZYD for 7 days, their bone marrow HSC were separated and counted. The phenotype Sca and cell cycle of HSC were detected by flow cytometer, and the colony formation was determined with semisolid methyl media culture.

RESULTSNo obvious difference in the number of mononuclear cell, suspended cell and colony production was found among all the groups (P > 0.05); while the expression of CD34 and Sca-1 increased in the low-dose XFZYD group, but in the middle-dose XFZYD group increase only showed in Sca-1 expression.

CONCLUSIONXFZYD plays a role of removing blood stasis and promoting regeneration through improving hematopoietic function by means of increasing the number and enhancing the function of premature HSC.

Animals ; Antigens, CD34 ; biosynthesis ; Antigens, Ly ; biosynthesis ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Count ; Cells, Cultured ; Colony-Forming Units Assay ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Male ; Membrane Proteins ; biosynthesis ; Mice ; Random Allocation

OBJECTIVETo investigate whether oxidized low-density lipoprotein (oxLDL) affects the survival and activity of endothelial progenitor cell (EPC) and whether the effects are mediated by lectin-like oxidized low-density lipoprotein receptor (LOX-1).

METHODSCD34+ cells isolated from human umbilical blood were cultured in endothelial cell growth medium-2 (EGM-2). After 14 days of culture, some EPCs were stimulated with 10, 25, 50 microg/ml of oxLDL for 48 hours; some were preincubated with LOX-1 mAb, a blocking antibody of LOX-1, for 24 hours, then exposed to 50 microg/ml oxLDL for 48 hours; others without any further treatment were used as control. The survival of EPC and the ability of adhesion, migration, and tube formation were examined. The levels of LOX-1 protein and mRNA expression were also assayed.

RESULTSIncubation with oxLDL at concentrations of 25 microg/ml or higher resulted in a dose-dependent increase of EPC apoptosis [25 microg/ml: (15.8 +/- 1.1.0%, 50 microg/ml: (18.8 +/- 2.0)% versus control: (9.0 +/- 1.2)%; P < 0.05]. Treated with oxLDL led to a significantly reduced migratry rate [25 microg/ml: (5.7 +/- 1.0)%, 50 microg/ml: (5.1 +/- 0.8)% versus control: (9.5 +/- 0.8)%; P < 0.05]. EPC treated with oxLDL showed a dose-dependent reduction of adhesion to fibronectin (25 Kg/ml: 33 +/- 2, 50 microg/ml: 30 +/- 3 versus control: 37 +/- 5; P < 0.05). Treatment with oxLDL impaired the in vitro vasculogenesis ability of EPCs. The total length of the tube structures in each photograph was decreased [25 microg/ml: (2.9 +/- 0.5) mm, 50 microg/ml: (1.8 +/- 0.5) mm versus control: (5.0 +/- 0.6) mm; P < 0.05]. The tube structure was severely disrupted, resulting in an incomplete and sparse tube network. However, all the detrimental effects on EPC were attenuated by pretreatment of EPC with LOX-1 mAb. In addition, Western blot analysis revealed that oxLDL increased LOX-1 protein expression from 100% to (172 +/- 8)% at a dose of 50 microg/ml. Furthermore, oxLDL caused an increase in LOX-1 mRNA expression from 100% to (174 +/- 39)% at a dose of 50 microig/ml.

CONCLUSIONOxLDL can directly inhibit EPC survival and activity and these effects are mediated by its receptor, LOX-1.

Antigens, CD34 ; metabolism ; Apoptosis ; Cell Adhesion ; Cell Movement ; Cell Survival ; Cells, Cultured ; Endothelial Cells ; drug effects ; physiology ; Fetal Blood ; cytology ; Humans ; Lipoproteins, LDL ; pharmacology ; physiology ; Neovascularization, Physiologic ; Scavenger Receptors, Class E ; biosynthesis ; physiology ; Stem Cells ; drug effects ; physiology

The purpose of this study was to investigate the prognosis of acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoid antigen-positive acute myeloid leukemia (Ly + AML), myeloid antigen-positive acute leukemia (My + ALL) and biphenotypic acute leukemia (BAL). Immunophenotyping was performed on medullary specimens of 197 acute leukemia (AL) patients by using three-color flow cytometry analysis and CD45/SSC gating. The scoring systems proposed by EGIL was adopted to classify the AL patients into five groups: 43 of ALL, 53 of AML, 53 of My + ALL, 39 of Ly + AML and 9 of BAL patients. The results showed that in Ly + AML, CD7 was the most common (53.8%) as compared to other lymphoid markers, however, in My + ALL CD13 was the most common (47.2%) as compared to other myeloid markers. Compared with Ly + AML, My + ALL had higher incidences of enlargement of liver, spleen and lymphnodes significantly (P<0.05). As for the case numbers of WBC counts > 100 x 10(9)/L, the positive rate of CD34 and the complete remission rate there was no obvious difference between groups of Ly + AML and My + ALL (P>0.05). As for incidences of enlargement of liver, spleen and lymphnodes, the case numbers of WBC counts > 100 x 10(9)/L, the positive rate of CD34 and complete remission rate, no obvious difference was found between ALL and My + ALL (P>0.05). Compared with AML, Ly + AML had lower complete remission rate significantly (P<0.05). As for incidences of enlargement of liver, spleen and lymphnodes, the case numbers of WBC counts > 100 x 10(9)/L and the positive rate of CD34, no obvious difference was found between AML and Ly + AML (P>0.05). Compared with Ly + AML and My + ALL, BAL showed no significant difference in complete remission rate (P>0.05) because the number of BAL patients was too small. It is concluded that since Ly + AML has lymphoid markers, and the prognosis of Ly + AML is worse than AML, the clinical therapy for Ly + AML should contain both AML and ALL. Though My + ALL had myeloid markers, no significant difference was found between My + ALL and ALL, it might be supposed that their therapy could be the same.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; biosynthesis ; Antigens, CD7 ; biosynthesis ; CD13 Antigens ; biosynthesis ; Child ; Child, Preschool ; Female ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; classification ; immunology ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; Prognosis

Mesenchymal stem cells (MSCs) have recently been identified and characterized in humans. Moreover, MSC secrete cytokines that can support hematopoietic progenitor growth. In the present study, we evaluated whether the efficacy of hematopoietic stem cell transplantation is improved by their co-transplantation with MSC, and whether this is positively correlated with the dose of infused MSCs. Accordingly, irradiated NOD/SCID mice were transplanted with 1x10(5) human CD34+ cells in the presence or absence of culture expanded MSCs (1x10(6) or 5x10(6)). We evaluated human hematopoietic cell engraftment by flow cytometry and assessed MSC tissue distributions by fluorescence in situ hybridization. We found that CD45+ and CD34+ cell levels were significantly elevated in a dose-dependent manner in cotransplanted mice 4 weeks after transplantation. The engraftments of CD33+ and CD19+ cells also increased dose-dependently. However, the engraftment of CD3+ cells did not increase after co-transplantation with MSCs. Human Y chromosome+ cells were observed in multiple tissues and were more frequently observed in mice co-transplanted with 5x10(6) rather than 1x10(6) MSCs. These results suggest that MSCs are capable of enhancing hematopoietic cell engraftment and distribution in multiple organs in a dose-dependent fashion.
Animals ; Antigens, CD34/*biosynthesis ; Cell Differentiation ; Cells, Cultured ; Dose-Response Relationship, Drug ; Female ; Fetal Blood/*metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Mesenchymal Stem Cells/*cytology ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Microscopy, Fluorescence/methods ; Stem Cell Transplantation/*methods

The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.
Adolescent ; Adult ; Antigens, CD34 ; Antineoplastic Agents ; pharmacology ; Bone Marrow Cells ; metabolism ; pathology ; Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Child ; Coculture Techniques ; Hematologic Neoplasms ; metabolism ; pathology ; therapy ; Humans ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Stromal Cells ; metabolism ; pathology ; Tretinoin ; pharmacology ; Tumor Cells, Cultured ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

This study was aimed to investigate the effect of some culture system composed of various cytokine combinations (TPO, SCF, FL, IL-1, IL-3, IL-6) on ex vivo expansion of megakaryocytic progenitors induced from CD34+ cells of peripheral blood and to seek a optimal cytokine combination and culture time. Mononuclear cells were isolated from mobilized peripheral blood (MPB) by density gradient centrifugation over Ficoll. CD34+ cells were purified by using an immunomagnetic bead separation system. The selected CD34+ cells were seeded in Iscove's modified Kulbecco's medium (IMDM) supplemented with fetal calf serum (FCS) and various kinds of cytokines. After 15 days of culture, the content of CD41+ cells in culture system were determined by flow cytometry, and the number of megakaryocyte colony-forming unit (CFU-MK) was measured simultaneously. The results showed that after definited days of culture, the cytokine combination TPO/FL/IL-6/IL-3 was the most suitable for MPB to obtain high number of MK, and better than any other three groups (P < 0.05). The increase multiple of CD41+ cells was 93.97 +/- 17.27 on day 5 and 131.23 +/- 18.26 on day 10. On day 15, the proportion and the increase multiple of CD41+ cells decreased obviously. The expansion multiples of CFU-MK were 93.33 +/- 10.02 on day 5 and 120.67 +/- 13.01 on day 10, higher than any other groups. It is concluded that TPO/FL/IL-6/IL-3 combination was the best optimal for expansion ex vivo of megakaryocytic progenitors from MPB, and its suitable duration of culture was 10 days; a culture system for expansion ex vivo of megakaryocytic progenitors have been established in this study.
Antigens, CD34 ; biosynthesis ; Blood Cells ; cytology ; Blood Platelets ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Interleukin-3 ; pharmacology ; Interleukin-6 ; pharmacology ; Megakaryocytes ; cytology ; Stem Cells ; cytology ; Thrombopoietin ; pharmacology

The aim of this study was to evaluate the homing capabilities of hematopoietic stem/progenitor cells (HSPCs) derived from human placenta tissues (PT). Single cell suspension of human PT was prepared by mechanical method. The expression levels of homing-related adhesion molecules (HRAM) including CD11a, CD49d, CD44, CD49e, CD62L and CD54 on CD34(+) cells and the percentages of CD34(+) cells and their subpopulations in nucleated cells (NC) from fresh human PT, umbilical cord arterial blood (UCAB) and umbilical cord venous blood (UCVB) were detected by using flow cytometry. The results showed that the percentage of CD34(+) cells and CD34(+)CD38(-) cells in placenta were higher than those in UCAB and UCVB. There were no significant difference in percentage of HSPC between UCAB and UCVB. Placenta-derived CD34(+) cells strongly expressed CD11a, CD49d, CD44, CD49e and CD54, among which expression levels of CD49e and CD54 on placenta-derived CD34(+) cells were significantly higher than those on UCAB and UCVB-derived CD34(+) cells. While the percentage of CD34(+)CD62L(+) cells in placenta was only lower than that in UCVB. It is concluded that human placenta is rich in HSPC. Moreover, the expression levels of most HRAM in CD34(+) cells from PT are higher than those from UCAB and UCVB or are close to them. It suggested that HSPCs derived from PT might have stronger homing capabilities than those from UCB.
Antigens, CD34 ; biosynthesis ; Cell Adhesion Molecules ; biosynthesis ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; metabolism ; Humans ; Hyaluronan Receptors ; biosynthesis ; Integrin alpha5 ; biosynthesis ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Placenta ; cytology