OBJECTIVETrying to find out the mechanism of microstructure influencing bacterial adhesion, we prepared different microstructures to compare the adsorptive behavior of graphite powder and adhesive behavior of oral microbe.

METHODSWe used polydimethylsiloxane (PDMS) to copy 23 microstructures of hydroxyapatite (HA) chip, and cultured them with different sizes graphite powder and oral microbes respectively, to observe and compare their behavior on microstructures.

RESULTSThe adsorption of 30-50 microm powder on different microstructures was insignificant, while 10-20 microm powder had maximum adsorption on 10 microm and 20 microm microstructures. Saccharomyces albicans was most likely to adhere to 5 microm microstructures which was equivalent to its own size. However, microstructures had little effect on adhesion of Streptococcus mutans which grew in a chain.

CONCLUSIONThe size of microstructure was the most effective factor that affects the adsorption of non-living powder, and it also had the same effect on the microbial adhesion; but some special bacteria, such as Streptococcus mutans which grew in a chain, was not affected by the sizes or shapes of microstructures.

Adsorption ; Bacteria ; Bacterial Adhesion ; Durapatite ; Graphite ; Mouth ; microbiology ; Streptococcus mutans

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.
Carcinoma, Hepatocellular ; immunology ; pathology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Killer Cells, Lymphokine-Activated ; cytology ; immunology ; Liver Neoplasms ; immunology ; pathology

This study was aimed to investigate the reversible effect of tetrandrine, toremifene and their combination on multidrug resistance of K562/A02 cell line. The IC(50) (the concentration causing 50% inhibition of cell growth) of adriamycin (ADR) were assayed by MTT method, the expression of MDR1 mRNA was measured by RT-PCR, the concentration of p-glycoprotein (P-gp) and intracellular ADR were detected by flow cytometry. The results showed that the IC(50) of ADR on K562/A02 and K562 cells were 57.43 and 1.16 mg/L, respectively. The IC(50) of ADR on K562/A02 cells after treatment with tetrandrine, toremifene and both combination were 14.12, 20.74 and 9.14 mg/L respectively, but both drugs did not influence the IC(50) of ADR on K562 cells. Pretreating K562/A02 cells with toremifene (2.5 micromol/L), tetrandrine (1 micromol/L) or both for 72 hours partially restored the sensitivity of K562/A02 cells to ADR. Tetrandrine and toremifene (alone or combination) elevated the ADR concentration in K562/A02, down regulated the expressions of P-gp and MDR1 mRNA. It is concluded that multidrug resistance of K562/A02 cells can be partially reversed by tetrandrine or toremifene, the combination of both drugs shows a higher synergistic reversal effect.
Antineoplastic Agents, Hormonal ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Benzylisoquinolines ; pharmacology ; Doxorubicin ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Toremifene ; pharmacology

The study was aimed to investigate the effective therapeutic method for patients with multiple myeloma accompanied with amyloidosis. A 58-year-old patient diagnosed as multiple myeloma accompanied with amyloidosis in four limbs was enrolled in this study. The various clinical and laboratorial examinations were performed, including bone marrow smear, immunologic test, radiography and so on. Patient received chemotherapeutic drugs and then autologous hematopoietic stem cell transplantation (auto-HSCT). The result showed that hematopoietic reconstitution was achieved at 23 days after auto-HSCT. Immunofixation electrophoresis was normal. There was only 0.6% plasma cells in the bone marrow. In conclusion, the auto-HSCT may be an effective treatment for multiple myeloma accompanied with amyloidosis in four limbs.
Amyloidosis ; complications ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; complications ; surgery ; Transplantation, Autologous

This study was aimed to investigate the reversible effect of cyclosporine A, raloxifene and their combination on multidrug resistance cell line K562/A02. The IC(50) (the concentration causing 50% inhibition of cell growth) of DNR were assayed by MTT method, the expression level of mdr-1 mRNA was assayed by RT-PCR, p-glycoprotein (P-gp) expression and intracellular DNR concentration were detected by flow cytometry. The results showed that the IC(50) of DNR on K562/A02 and K562 cells were 23.51 mg/L and 0.29 mg/L, respectively. The IC(50) of DNR on K562/A02 cells in treatment with raloxifene CsA and both combination were 5.98, 8.15 and 3.68 mg/L respectively, but both drugs not influenced IC(50) of DNR on K562 cells. Pretreating K562/A02 cells with raloxifene (2.5 mg/L) or CsA (1 mg/L) for 48 hours partially restored the sensitivity of K562/A02 cells to DNR. Cyclosporine A and raloxifene (alone or combination) elevated the intracellular DNR concentration in K562/A02, down regulated P-gp and mdr-1 mRNA expressions. It is concluded that multidrug resistance (MDR) can be partially reversed by CsA or raloxifene, the combination of both drugs shows a great synergistic reversal effect.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; drug effects ; genetics ; Cyclosporine ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; drug effects ; pathology ; RNA, Messenger ; biosynthesis ; drug effects ; genetics ; Raloxifene Hydrochloride ; pharmacology

OBJECTIVETo investigate the reversible effect of nilotinib, BrTet (5-bromotetrandrine) and their combination on multidrug resistance cell line K562/A02 and its mechanism.

METHODSCell proliferation inhibition was assessed by MTT method and cell apoptosis by flow cytometry (FCM). The expression of mdr1 mRNA was determined by RT-PCR, and the expression of P-gp was assessed by Western blot.

RESULTSAfter 48 h 5 nmol/L nilotinib or 0.5 µmol/L BrTet treatment, IC(50) of daunorubicin (DNR) to K562/A02 was 4.52 mg/L or 5.41 mg/L respectively; While on combinative treatment, its IC(50) decreased to 2.98 mg/L. Nilotinib or BrTet alone was not able to increase the DNR induced apoptosis rate of K562/A02 cell (P > 0.05), while on combination treatment the apoptosis rate increased remarkably. After 48 h 5 nmol/L nilotinib or 0.5 µmol/L BrTet treatment alone, gray-scale value of mdr1 mRNA was 0.48 ± 0.04 or 0.64 ± 0.01, respectively; while on combinative treatment the value decreased to 0.35 ± 0.04. The P-gp expression level in K562/A02 cells was 0.61 ± 0.05, or 0.52 ± 0.02 when treated with 5 nmol/L nilotinib or 0.5 µmol/L BrTet alone for 48 h, but on combination treatment, the level decreased to 0.44 ± 0.03.

CONCLUSIONNilotinib or BrTet alone can partially reverse drug resistance of K562/A02 cells. The mechanism may be associated with the decrease of mdr1 mRNA and P-gp expression and increase of the apoptosis rate. And there is a synergistic action with these two agants in combination.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells

This study was aimed to establish a cytokine-independent human myelodysplastic cells line from bone marrow of a patient with MDS-CMML. This cell line was incubated in mixed culture of RPMI 1640 and DMEM with 15% bovine serum, but without cytokines; its biological characteristics were identified by morphology, surface marker profiles, cell proliferation, differentiation and apoptosis. The results showed that the established cell line could not depend on cytokines for long-term survival and growth, and could differentiate into colony-forming unit-macrophage, colony-forming unit-megakaryocyte. In conclusion, a cytokine-independent human myelodysplastic syndrome cell line, named MDS-JSN04 (MDS Nanjing Jiansu 04), was established. Its partial biological characteristics were identified and clarified.
Antigens, CD19 ; analysis ; Antigens, CD20 ; analysis ; Apoptosis ; drug effects ; Bone Marrow Cells ; metabolism ; pathology ; ultrastructure ; CD79 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Culture Media ; pharmacology ; Cytokines ; pharmacology ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Humans ; Lewis X Antigen ; analysis ; Male ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Sialic Acid Binding Ig-like Lectin 2 ; analysis ; Time Factors

The study was aimed to investigate the possible mechanism of vitamin K(2) (VK(2)) on myelodysplastic syndrome (MDS) cell line MUTZ-1 in vitro. The flow cytometry was used to analyze apoptosis rate and the change of cell cycle. The expression of apoptosis-related genes bcl-2, survivin and bax were detected by reverse transcription-polymerase chain reaction (RT-PCR). The activity of caspase-3 was detected by chemiluminescence assay. The results indicated that the apoptosis peak on FCM and positive Annexin-V FITC on cell membrane showed that VK(2) induced apoptosis of MUTZ-1 cells in a dose-and-time-dependent manner, S and G(2) cell decrement, G(0)/G(1) cell arrest, VK(2) significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax, the activity of caspase-3 was significantly increased. It is concluded that VK(2) induces apoptosis of MUTZ-1 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2 and survivin may play important roles in the process of apoptosis induction.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Myelodysplastic Syndromes ; drug therapy ; pathology ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Vitamin K 2 ; pharmacology ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo study the effect of dendritic cells (DC) co-cultured with cytokine induced killer (CIK) cells on cytotoxicity against K562 and K562 drug-resistant (K562/ADM) cells.

METHODSPeripheral blood mononuclear cells (MNC) isolated from healthy adult donors were induced to obtain CIK cells and DC respectively and then these two kinds of cells were co-cultured. The cytotoxicity of the co-cultured cells against K562 and K562/ADM cells was measured with MTT assay.

RESULTSThe cytotoxicity CIK cells alone to K562 and K562/ADM cells was (20.0 +/- 1.2)% - (61.1 +/- 2.2)% and (17.5 +/- 2.1)% - (45.2 +/- 3.3)% respectively at low effector to target ratios (2.5 - 20.0). This effect was significantly enhanced by co-culturing with DCs being (25.2 +/- 2.3)% - (70.9 +/- 4.1)% and (22.4 +/- 2.7)% - (62.3 +/- 5.0)%.

CONCLUSIONCIK cells showed high cytotoxicity against K562 and K562/ADM cells and the activity could be enhanced by co-culturing with DC.

Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Humans ; Immunophenotyping ; K562 Cells

The aim of this study was to analyze the hematopoietic chimerism after non-myeloablative allogeneic peripheral blood stem cell transplantation (NAPBSCT). 28 patients received NAPBSCT were evaluated. The conditioning regimen included FBC (fludarabine, busulphan, cyclophosphamide) +/- Ara-C. Peripheral blood was collected before and after transplantation in different periods. Semi-quantitative assessment of hematopoietic chimerism was performed by short tandem repeat-polymerase chain reaction (STR-PCR), polyacrylamide gel electrophoresis (PAGE) and silver staining, and analyzed by Image Analysis System. The results showed that on day 30 after transplantation, one patient failed to engraft, but 22 cases formed complete chimerism (CC) and 5 cases were of mixed chimerism. On day 7 after transplantation, the average percentage of donor cells was 74.71%. The time of dominance of the donor-specific allelic pattern preceded the recovery time of neutrophils and platelets. The incidence of aGVHD in group CC was significantly higher than that in group MC (P < 0.05). There was no significant difference in the incidence of cGVHD and disease relapse between group CC and group MC (P > 0.05). One patient relapsed in CC status without a transitional stage of MC. One patient with MC rejected grafts in early stage. 3 patients with MC transferred to CC and got complete remission after early implementation of therapy. It is concluded that sequential and quantitative detection of chimerism may be of great value to evaluate engraftment and to predict graft rejection, disease relapse and GVHD. Furthermore, it may provide a basis for early intervention treatment in the related complications.
Adolescent ; Adult ; Chimerism ; Female ; Graft vs Host Disease ; prevention & control ; Humans ; Leukemia, Myeloid, Acute ; therapy ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Transplantation Chimera ; blood ; genetics ; Transplantation Conditioning ; Transplantation, Homologous