Aged ; Biopsy ; Bone Marrow Examination ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphoma, B-Cell ; diagnosis ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To investigate the effects of transplantated tumor capillary permeability under microbubble enhanced diagnostic ultrasound exposure.Methods Fourty eight rats were randomly divided into four groups after Walker 256 carcinomas were subcutaneousely implanted,named the control group (A),microbubbles group (B),ultrasound group (C),microbubbles and ultrasound group (D).Using Evans blue(EB) as the indicator for tumor capillary permeability,the microbubbles were injected through the vein with diagnostic ultrasound frequency as 1.75 MHz,mechanical index as 1.4.The spillover of EB was estimated under confocal laser microscope,meanwhile the contents of EB in the tumor tissues were calculated according to the standard curve and spectrophotometry.Results EB spillover was observed around the capillaries in the group D,while EB spillover was only observable on capillary in group A,B and C.The EB content was also significantly higher than those in group A,B and C.Conclusions Exposure to both diagnostic ultrasound and microbubbles results in increased capillary permeability of rat tumor tissues,which might become a potential therapeutic method on clinical chemotherapy.

AIM:To detect the treatment of K562 leukemia cells with bortezomib altering the expression of genes fas,bcl-2,bcl2l12,bim,bax,caspase-9 and caspase-3.METHODS:MTT assay was used to detect the inhibition of proliferation.Apoptosis was detected by Annexin-V staining and mitochondrial transmembrane potential(??m).RT-PCR was used to analyze the mRNA expressions of fas,bcl-2,bcl2l12,bim,bax,caspase-3 and caspase-9.RESULTS:Bortezomib caused a time-and dose-dependent inhibition of cell proliferation and IC50 of 24 h and 48 h were 161.41 nmol/L and 96.33 nmol/L,respectively.At the concentration of 104 nmol/L,bortezomib induced apoptosis in a time-dependent manner,including increasing annexin-V positivity and decreasing the ??m.RT-PCR showed that bortezomib up-regulated the mRNA expression of fas,bcl2l12,caspase-9 and caspase-3,but mRNA expressions of bcl-2,bim and bax did not changed obviously.CONCLUSION:Bortezomib inhibits the proliferation of K562 and induces apoptosis,in which fas,bcl2l12,caspase-9 or caspase-3 gene is one of the main genes taking part in.

Objective To evaluate the growth inhibition of human gastric carcinoma cell lines SGC 7901 in vitro and the expression of bcl-2, bcl 2l12 and bax with docosahexaenoic acid (DHA) and 5-fluorouracil (5-FU). Methods The effect of DHA and 5-FU was measured by trypan blue, and the interaction between two agents was judged by combination index (CI). Cells were observed by inverted microscope. Flow cytometry was used for analysis of apoptosis by PI staining and Annexin-V/PI. RT-PCR was used to analyze the levels of bcl-2, bcl 2l12 and bax mRNA. Results DHA significantly inhibited the growth of SGC 7901 cells in a dose- and time-dependent way ( P < 0. 05 ), the IC50 of 24 h and 48 h was 67. 81 μg/ml and 45.76 μg/ml, and a strong synergism was found in the combination of DHA and 5-FU (CI < 1 ,P <0. 01 ). Treated by DHA and 5-FU for 48 h, cells became sparse under inverted microscope. DHA or 5-FU was able to induce apoptosis and the effect became even more significant by the combination of DHA and 5-FU. Cells were holted in phase of G01/G1 and S. RT-PCR showed that DHA or 5-FU down-regulated the expression of bcl-2 and bcl 2l12 mRNA, while bax mRNA expression was not downregnlated. Conclusions DHA could inhibit the growth of gastric carcinoma cells, DHA and 5-FU had synergetic effect in the inhibition of the cells growth and blockage of the cell cycles possibly by down-regulating the expression of bcl-2 and bcl 2l12.

Many studies indicate that lymphoid neoplasms are related with chromosome translocations and the molecular alterations involving in the cell cycle and/or apoptotic pathways. However, survival of B and T tumor cells also depends on interactions of these cells with the accompanying cells comprising the lymphoma microenvironment. Immune cells, stromal cells and numerous molecular together make up the microenvironment and have functional interaction with tumor cells, promoting tumor growth and drug resistance. Different types of lymphoma have various clinical courses, therapy responses and prognoses, which show a close relationship with the microenvironment. This review summarizes several components of lymphoma microenvironment including macrophages, adhesion molecules and chemokines and the roles of microenvironment in classic non-Hodgkin's lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, suggesting that the microenvironment influence the prognosis of lymphoma, targeting microenvironment may be a potential method in lymphoma therapy.
Apoptosis ; Cell Adhesion Molecules ; Cell Cycle ; Chemokines ; Humans ; Lymphoma ; metabolism ; pathology ; Macrophages ; Tumor Microenvironment

OBJECTIVETo explore a new method for in vitro expansion of human mesenchymal stem cells (MSC) by using platelet-rich plasma (PRP) in place of fetal calf serum (FCS).

METHODSBone marrow(BM) samples were obtained from the proximal femurs of patients with normal haematopoietic function undergoing total hip arthroplasty. 2 x 10(5)/cm2 BM nucleated cells were seeded in 25 cm2 flasks for MSC cultivation containing one of the 3 mediums: complete Dexter medium with 12.5% FBS and 12.5% horse serum (medium1), alpha-MEM with 10% FCS (medium2) and alpha-MEM with 5% PRP(medium3). At the same time, 1 x 10(6) nucleated BM cells and same amount of nucleated cells from iliac aspirate were seeded in 25 cm2 tissue flasks, colony forming unit-fibroblast (CFU-F) assay.

RESULTSCulture-expanded cells from proximal femurs presented a typical fibroblast-like morphology. Cells were positive for SH2 (CD 105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. On induction, these cells could differentiate into osteoblasts. A significantly higher proliferative capacity of MSCs expanded in medium3 was observed in comparison to those in mediuml or 2 without alteration of the phenotype and the differentiation property. CFU-F assays indicated that bone marrow from the proximal femoral contained significantly more CFU-F than that from iliac aspirate.

CONCLUSIONPlatelet-rich plasma can be used in place of FCS to provide a safer and more effective culture condition to expand MSCs for clinical purpose. The proximal femur BM cells can be obtained in hip surgeries.

This study was aimed to investigate the expression level of stromal cell derived factor-1 gene (SDF-1) in bone marrow mesenchymal stem cells (MSC) of patients with myelodysplastic syndrome (MDS). The MSC from bone marrow samples of MDS patients were isolated, cultured and expanded, the morphology and immunophenotype of MSC were analyzed. The expression levels of SDF-1 and internal reference GAPDH in MSC of MDS patients were detected by real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method and were compared with expression levels of healthy donors. The results showed that the expression levels of SDF-1 in MDS patients were significantly different from those in healthy donors (1.53 +/- 0.92 vs 5.51 +/- 0.99) (P < 0.01). SDF-1 gene expression levels in bone marrow MSC of MDS patients were significantly higher than that in MSC derived from healthy donors. It is concluded that the abnormal expression of SDF-1 gene in MSC may influence the regulation of hematopoiesis of the bone marrow microenvironment in MDS patients and it is worthy of further investigation for new clue on etiological mechanism and treatment of MDS.
Adult ; Aged ; Bone Marrow Cells ; metabolism ; pathology ; Cells, Cultured ; Chemokine CXCL12 ; biosynthesis ; genetics ; Female ; Humans ; Male ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology

To study the biological characteristics of mesenchymal stem cells (MSC) from patients with myelodysplastic syndrome (MDS) and their supportive capacity for hematopoiesis in vitro, MSCs from bone marrow samples of MDS patients were isolated, cultured and expanded. Morphology, immunophenotype, osteoblasts differentiative and proliferative property of MSC and colony forming unit-fibroblast (CFU-F) were measured and analyzed. Mononuclear cells (MNC) of cord blood were plated onto a feeder layer formed by MSC of MDS patient, cells count and CFU-GM production were observed. The results showed that the culture-expanded cells from MDS patients presented a typical fibroblast-like morphology. Cells were positive for SH2 (CD105), SH3 (CD73), Thy-1 (CD90), but negative for CD34 and CD45. After induction, these cells could differentiate into osteoblasts. Their proliferative capacity and CFU-F number were similar to those of MSC from healthy donors. The total cell count and CFU-GM yield in supernatants after culture for 2 weeks were significantly lower than those of control in hematopoiesis supportive experiments in vitro (P < 0.05). It is concluded that the biological characteristics of MSC from bone marrow of MDS patients are not different from those of MSC isolated from bone marrow of normal donors, however, their capacity of hematopoiesis support in vitro are significantly weaker.
5'-Nucleotidase ; analysis ; Adult ; Aged ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; immunology ; Cell Differentiation ; Endoglin ; Female ; Hematopoiesis ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; immunology ; Middle Aged ; Myelodysplastic Syndromes ; blood ; Receptors, Cell Surface ; analysis

Objective To study the changes of the peripheral blood regulatory B cells and related cytokines in children patients with chronic idiopathic thrombocytopenic purpura (CITP),and to analyze their correlations with effective Th cells,and to discuss their roles in the pathogenesis of CITP.Methods The peripheral blood mononuclear cells (PBMC)were separated in 30 cases of CITP,then percentages of Th1 ,Th17,Th22 and Breg cells were detected by the flow cytometry(FCM).IFN-γ,IL-17,IL-22 and IL-10 levels in culture supernatant were measured by the ELISA method.Results Compared with the control group,the proportions of the peripheral blood Th1 ,Th17,Th22 subsets in CITP children were increased[(18.4±4.7)% vs.(10.82±4.4)%;(2.42±1.02 )% vs.(1 .23±0.42)%;(0.79±0.24)% vs.(0.26±0.11)%],the proportion of Breg cells was reduced significantly,the differ-ences had statistical significance[(0.83±0.21)% vs.(1 .89±0.12)%,P <0.05].In CITP children,the IFN-γ,IL-17,IL-22 levels in culture supernatant were higher than those in the control group[(278.2±121.2)pg/mL vs.(181.8 ±82.2)pg/mL;(214.8 ± 100.5)pg/mL vs.(161 .4±67.8)pg/mL;(122.16±22.2)pg/mL vs.(90.8±31.1)pg/mL],but the IL-10 level was lower than that in the healthy control group[(27.4±12.6)pg/mL vs.(46.1±16.2)pg/mL),differences between them had statistical signifi-cance(P <0.05).Besides,there was a positive correlation between the proportion of Breg cells and cytokine IL-10 level(r=0.617, P <0.05 ),however,there was a negative correlation between the proportion of Breg cells with Th1 ,Th17 and Th22 cells (P <0.05),between IL-10 level with IFN-γ,IL-17 and IL-22 levels(P <0.05).Conclusion The downregulation of Breg cells proportion may participate in the immune disorder mechanism of effective Th cells in CITP children,which can provide a new idea for the im-munoregulation therapy in CITP children.

ObjectiveTo investigate the gene expressions of TAZ and Wnt/β-catenin on the postosteogenic cells of mesenchymal stem cells(MSC)in multiple myeloma(MM)patients and to explore the potential therapeutic target of multiple myeloma bone disease (MBD).MethodsBone marrow mononuclear cells MNC from MM and controls were isolated,cultured,expanded and then induced to osteogenic differentiation.Realtime quantitative RT-PCR was employed to detect the osteogenic markers (TAZ,Wnt/β-catenin,OPN,OC,ALP and Cbf α1); and alizarin red staining for mineral deposition.The mRNA expressions of TAZ and Wnt/β-catenin in the two groups were analysed.ResultsAlizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC.The mRNA expressions of OC,ALP and Cbf α1 were 2.0958±0.5665,2.6670±0.3847,0.8463±0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly higher than those of pre-osteogenic cells(1.3487±0.9291,1.1452±0.6054,0.4439±0.2945) (t =2.171,6.709,2.795; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP and Cbf α1 were 2.1096±0.8267,2.8991±0.3531,4.3045±0.2844,1.3273±0.4075,respectively,on the post-osteogenic cells of MSC in the controls,which were significantly higher than those of pre-osteogenic cells (1.2200±0.9091,0.8780±0.3927,1.9161±0.2684,0.6736±0.2513) (t =2.289,12.103,25.134,4.411; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP,Cbf α1 were 1.2710±0.5636,2.0958±0.5665,2.6670± 0.3847,0.8463+0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly lower than those of control groups(2.1096 ±0.8267,2.8991 ±0.3531,4.3045±0.2844,1.3273±0.4075) (t =-2.650,-3.805,-10.822,-2.841; all P ＜ 0.05).The mRNA expression of TAZ and β-catenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic cells of MSC in MM patients,which were significantly lower than those of control groups (4.4140±0.8325,0.9516±0.2920) (t =±5.085,-3.235;both P ＜ 0.05).ConclusionThe gene expressions of OPN,OC,ALP and Cbf α1,the osteogenesis related genes,are increased in post-osteogenic cells of MSC,which showed the MSC have been successfully induced to osteoblasts.Comparing with control groups,the osteogenic potential of MSC in MM patients is lower.Based on the above research,TAZ and Wnt/β-catenin may present a novel target for the future therapy of MBD.