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

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

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

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

This study was purposed to explore the effect of different cytokine combinations on the expansion of the mononuclear cells drived from umbilical cord blood (CB) ex vivo and expression of CXCR4 and CD49d on CD34+ cells after expansion. Human fresh CB mononuclear cells were cultured in serum-free and stroma-free medium containing different combinations of cytokine for 7 days. At day o and 7, the total cells were counted, CD34+ cells and CD34+CXCR4+, CD34+CD49d+ cells were assayed by flow cytometry, and CFU were determined. According to the different combinations of cytokine, experiments were divided into four groups: control, SF group (SCF + FL), SFT group (SCF + FL + TPO) and SFT6 group (SCF + FL + TPO + IL-6). The results showed that the SF (SF group) combination supported only low expansion of total cells, CD34+ cells and CFU. The addition of TPO in SF group restored UCB stem/progenitors expansion to a higher level than that in SF group, while there was no difference between groups SFT and SFT6 (P > 0.05). The cytokine combinations in groups SF, SFT and SFT6 all could upregulate the expression levels of CD49d and CXCR4 on expanded cord blood CD34+ cells, but there were no significant differences between groups SF, SFT and SFT6 (P > 0.05). It is concluded that SCF + FL has no strong synergistic effects on primitive hematopoietic cells. TPO plays an important role in enhancing expansion of umbilical cord blood hematopoietic cells, while IL-6 only shows a neutral effect on it. SCF + FL + TPO combination not only promotes progenitor cells expansion but also upregulates the expression of CD49d and CXCR4 on CD34+ cells from cord blood.
Antigens, CD34 ; biosynthesis ; genetics ; Cytokines ; pharmacology ; Drug Synergism ; Fetal Blood ; cytology ; Humans ; Integrin alpha4 ; biosynthesis ; genetics ; Leukocytes, Mononuclear ; cytology ; Membrane Proteins ; pharmacology ; Receptors, CXCR4 ; biosynthesis ; genetics ; Stem Cell Factor ; pharmacology ; Thrombopoietin ; pharmacology

To evaluate soluble GM-CSF-Ralpha expression in patients with acute myeloid leukemia (AML) and its clinic significance, plasma concentration of solGM-Ralpha in de novo 66 patients with AML was detected by enzyme-linked immuno-sorbent assay, and the relationship between solGM-Ralpha levels and various clinical parameters was analyzed. The result showed that the levels of solGM-Ralpha in plasma of patients with AML were significantly higher than that in plasma of normal controls; the lowest level of solGM-Ralpha was found in plasma of patients with AML-M3 (3897.75 +/- 2651.43 pg/ml), the highest level of solGM-Ralpha was observed in plasma of patients with AML-M5 (9990.92 +/- 6325.43 pg/ml). Patients with high level of solGM-Ralpha were generally accompanied with a distinct clinical picture, including higher counts of white blood cell and myeloid precursors, as well as higher expression of CD34, CD95 and CD116 antigen. It is concluded that the high level of solGM-Ralpha in plasma of patients may suggest AML poor prognosis and play a role in pathogenesis of leukemia, the GM-CSF and its receptor solGM-Ralpha needs further study.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Leukemia, Myeloid, Acute ; blood ; Male ; Middle Aged ; Prognosis ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; blood ; fas Receptor ; blood

HOXB4, a member of homeobox gene family, is closely related to the self-renewing and proliferative ability of primitive hematopoietic stem/progenitor cells (PHSC/PHPC). This study was aimed to investigate the self-renewing level of cord blood progenitor cells (CBPC) expanded in vitro. The HOXB4 expression at mRNA level was assayed by using real time RT-PCR. The results indicated that as culture prolonged, the total cells, CD34(+) cells greatly increased, however the HOXB4 expression gradually declined, even down to undetectable level similar to that of mature lymphocytes. Meanwhile, it was shown that CD34(+) cells co-cultured with bone marrow mesenchymal stem cells (BM-MSC) could abate the decline of HOXB4 expression. It is concluded that the self-renewing potential of CD34(+) cells gradually decreased during expansion in vitro, co-culture with BM-MSC was helpful to CD34(+) cell expansion and slowed the loss trend of its self-renewal.
Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Homeodomain Proteins ; biosynthesis ; genetics ; Humans ; Mesenchymal Stromal Cells ; cytology ; Transcription Factors ; biosynthesis ; genetics