0.5ug/ml in 34 patients(79.1%).Postoperative survival rate and recover of the work ability in group T were significantly higher than those in group C.Conclusions EPTT before RT for the type I diabetes patients with renal disorder can improve the results of RT.

Objective To study the pathogenic features, diagnosis and treatment of aberrant thyroid cancer.Methods A retrospective analysis of clinical and pathological data of 29 cases of aberrant thyroid cancer was made.Results All of the 29 patients underwent operative treatment and postoperative adjuvant radiation therapy and chemotherapy. On postoperative follow up, the 5-year survival rate was 52.0%. The longest survivor patient was alive 24 years after operation.Conclusions The key to increase the survival rate of patients is early detection and timely surgical treatment. Postoperative adjuvant radiation therapy and chemotherapy are conducive to increase survival rate.

Objective:To evaluate the micro-push-out bond strengths of prefabricated glass fiber posts with poly-dopamine functionalized to root dentin using resin cements, contrasted with silane treatment. Methods:In the study, 30 glass fiber posts were randomly divided into 3 groups ( 10 posts in each group) for different surface treatments. Group 1, treated with poly-dopa; Group 2, treated with silane coupling agent for 60s;Group 3, no surface treatment (Control group). The 30 extracted human, single-rooted teeth were endodontically treated and a 9 mm post space was prepared in each tooth with post drills provided by the manufacturer. Following post cementation, the specimens were stored in distilled water at 37 ℃ for 7 days. The micro-push-out bond strengths were tested using a universal testing machine (0. 5 mm/min), and the failure modes were examined with a stereomicroscope. The data of the three groups were statistically analyzed using the one-way ANOVA test(α= 0. 05). Results:The bond strengths were (7. 909 ± 1. 987) MPa for Group 1, (5. 906 ± 0. 620) MPa for Group 2, and 4. 678 ± 0. 910 MPa for Group 3 . The bond strength of poly-dopamine group was significantly higher than that of the silane group (P<0. 05). Conclusion: Contrasted with silane treatment, surface poly-dopamine functionaliza-tion was confirmed to be a more reliable method for improving the bond strength of resin luting agents to fiber posts.

OBJECTIVETo explore the effects of rhEPO on the migration of bone marrow(BM) derived mesenchymal stem cells(MSCs) and its probable signal transduction mechanism.

METHODSMSC was cultured by classical whole BM adherence method; MSC characteristics was identified by multi-differentiation and surface marker (CD90, CD133, CD34, CD105). The effect of different concentrations EPO (1, 10, 100, 1000 IU/ml) on MSCs migration were observed. Then 30 minutes later, MSC were treated with signal transduction pathway inhibitors, 50 nmol/L wortmannin, 50 micromol/L PD98059, 10 micromol/L U73122, 4 microg/ml Anti EPO-R IgG, 30 micromol/L SB203580, 10 mmol/L Staurosporine, 6 nmol/L G06976 and 50 micromol/L Pseudo Z, respectively. The efficacy of MSC migration was analyzed by Transwell in vitro migration assay.

RESULTSCultured MSCs had the capacities for osteogenic and adipogenic differentiation and highly expressed CD105, CD90 and EPO-R. The efficiency of MSC in vitro migration increased gradually in a concentration-dependent manner with increasing concentration of rhEPO, and the ability peaked at a concentration of 100 IU/ml. Furthermore, the migration ability was decreased on treated with U73122, Anti EPO-R IgG, Staurosporine, Pseudo Z treatment.

CONCLUSIONEPO/EPO-R-mediated MSCs migration is related with MAPK, PI-PLC/PKC-zeta signal pathways, PKC-zeta signal pathway may be of central role for MSCs migration.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Erythropoietin ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar ; Recombinant Proteins ; Signal Transduction ; drug effects

This paper was designed to investigate the effect of laminar shear stress on matrix metalloproteinase -9 (MMP-9) expression in rat bone marrow-derived mesenchymal stem cells (MSCs), and the possible signal transduction mechanism involved. Rat bone marrow MSCs were isolated and cultured, then, exposed to laminar shear stress at indicated strengths such as low (5dyne/cm2), medium (15 dyne/cm2) and high (30 dyne/cm2) via parallel plate flow chamber. RT-PCR was used to analyze the expression of MMP-9. The signaling inhibitors such as Wortmannin (PI3K specific inhabitor), SB202190 (p38MAPK specific inhabitor), and PD98059 (ERK1/2 specific inhabitor) were used to investigate the possible mechanical signal transduction pathway. The results showed: (1) The expression of MMP-9 was weak in static state, however, MMP-9 expression increased when MSCs were exposed to 15 dyne/cm2 shear stress for 2 hours, and MMP-9 expression increased with the extension of stimulating time, and it reached the peak at 24 h; (2) MSCs were stimulated by shear stress for 2 hours at different strengths (5 dyne/cm2, 15 dyne/cm2, 30 dyne/cm2), and under all these conditions, the expression of MMP-9 increased, and reached the peak at 15 dyne/cm2; (3) After MSCs were pretreated by three kinds of signal pathway inhibitors, the expression of MMP-9 did not change obviously in Wortmannin group and PD98059 group, but it was significantly inhibited in SB202190 group. This study demonstrated that shear stress could induce the expression of MMP-9 in rat bone marrow-derived mesenchymal stem cells; the amount of MMP-9 expression was closely related to stimulating time and the strengths of shear stress; and p38MAPK signal pathway played a critical role during the process.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cells, Cultured ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Rats ; Signal Transduction ; Stress, Mechanical

To evaluate the effects of rhG-CSF on mobilization of mesenchymal stem cells (MSCs) of mouse bone marrow at different time point, thirty mice were randomly divided into rhG-CSF treatment group and control group. The mice were subcutaneously injected with rhG-CSF in a dose of 80 microg/kg or saline for 5 days. The bone marrow and peripheral blood were obtained at time points of 6, 12, 168 hours after final injection of rhG-CSF or saline. Bone marrow mononuclear cells (BMMNCs) were seeded at density of 1 x 10(6) MNCs onto 12-well plate for culture expansion in DMEM supplemented with 10% FBS, and the number of colony forming unit - fibroblast (CFU-F) was counted after 14 days. The cells were collected by trypsinization and the surface antigens CD34, CD133, CD90 and CD105 were analyzed by flow cytometry. The multi-differentiation of MSCs were done in the culture condition of induced-adipocyte and osteocyte. Peripheral blood MNCs examination was same as the bone marrow. The results indicated that the number of CFU-F of bone marrow in rhG-CSF group was more than that in control group (p < 0.01), the number of CFU-F in rhG-CSF group at 6 hours was more than that at 12 hours and 168 hours, respectively (p < 0.01). There was no obvious difference between CFU-F at 12 hours and at 168 hours (p > 0.05). MSCs were positive for CD90, CD105 and negative for CD34 and CD133. MSCs were found to differentiate into adipocyte and osteocyte in vitro. The CFU-F of PBMNCs obtained and cultured in vitro in the same culture conditions could be observed after the rhG-CSF injection at 6 hours, but cloning efficiency was (0.50 +/- 0.11) x 10(-6) MNCs and showed statistical difference as compared with control. It is concluded that rhG-CSF to mobilize hemopoietic stem cells can be used to induce mouse MSCs in vivo expansion, which showed the peak value within 6 hours after final injection of rhG-CSF. rhG-CSF have the mini-mobilization effect on murine MSCs derived from bone marrow.
Animals ; Cells, Cultured ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Mesenchymal Stromal Cells ; cytology ; Mice ; Random Allocation ; Recombinant Proteins

To investigate the inhibitory effect of sildenafil on Caco-2 cell proliferation and its anti-inflammatory effect on menadione-induced NCM460 cell inflammation model, MTT assay was used to determine cell proliferation. Intracellular reactive oxygen species(ROS)and nitric oxide(NO)levels were detected by fluorescent probe. Western blot was used to detect the expression of eNOS/ERK/JNK pathway related proteins in Caco-2 cells and correlated inflammatory cytokines in NCM460 cells. The effect of sildenafil on the growth of two probiotics was determined by spectrophotometry. Results showed that sildenafil signi-ficantly inhibited the proliferation of Caco-2 cells and enhanced the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins in Caco-2 cells; while after adding NG-nitro-L-arginine methyl ester(L-NAME), the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins were significantly lower than those of the sildenafil group. Compared with the menadione group, sildenafil significantly reduced ROS levels in NCM460 cells and inhibited the expression levels of IL-6, IL-1β, p62, and TNF-α. Moreover, high concentrations of sildenafil had no obvious toxic effects on Lactobacillus casei and Lactobacillus rhamnosus. Thus, the results indicated that sildenafil could effectively inhibit the intestinal inflammatory response without affecting the balance of the intestinal flora, and prevent colorectal cancer by reducing the oxidative stress responses in the intestinal cells.

This study was aimed to investigate the effects of recombinant human erythropoietin (rhEPO) on proliferation of human bone marrow-derived mesenchymal stem cells (MSCs) in vitro. The aspirates of the bone marrow from healty volunteers were seeded in culture medium. Then MSCs were isolated according to characteristics adhering to the plastics. After three passages in culture, bone marrow-derived adherent cells were identified by growing morphological features, cell surface antigens and differentiation into multi-lineages. Then P3-MSCs which had been identified were incubated with different concentrations of rhEPO (0.5, 1, 5, 10 and 50 U/ml). Subsequently, proliferation of MSCs was measured by MTT assay, as well as cell counts. At the same time, cell cycle was detected by flow cytometry (FCM). The results indicated that the expressions of CD90 and CD105 in P3 bone marrow-derived adherent cells were positive, while the expressions of CD34 and CD45 were negative, and these cells could differentiate into adipocytes, osteocytes and chondrocytes in induction media. MTT assay showed that the optical density (OD) of group treated with EPO was significantly higher than that in the control group (p<0.05), and the group treated with 50 U/ml EPO achieved the most predominant effects. The results of cell count were coincident with that of MTT assay. Furthermore, the cell cycle analysis by FCM revealed that rhEPO could relatively decrease the cell ratio in G0/G1 phase, and increase the cell ratio in S and G2/M phases. As compared with the control group, all those differences were statistically significant (p<0.01). It is concluded that erythropoietin can promote proliferation of human bone marrow mesenchymal stem cells in vitro, which may be correlated with the increased entry into S and M phases of cell cycle of MSCs adjusted by EPO.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Culture Media ; Erythropoietin ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Recombinant Proteins

AIMTo observe the effect of angiotensin-converting enzyme inhibitors (ACEI) and aldosterone receptor blockers on cardiac function to explore the mechanism of cardiac function descending and myocardial injury in calcium-overload rats.

METHODSCalcium-overload in rat was induced by administration of Vitamin D3 plus nicotine. To Estimate the extent of calcium-overload by calcium content. Angiotension II and aldosterone levels in the myocardia were measured by radioimmunoassay. Cardiac function (+/- LVdp/dt, LVESP and LVEDP) were measured by Powerlab. The malondialdehyde (MDA) content, activities of lactate dehydrogenase (LDH) and creatine kinase (CPK) were measured by biochemistry.

RESULTSCalcium content increased by 3.2-, 5.8 -fold in myocardial and artery, compared with controls. VDN-treated survivors showed lower + LVdp/dt(max) and -LVdp/dt(max) values, by 27% and 34%, respectively (both P < 0.01). Higher LVESP, and LVEDP by 42 % and 32% (P < 0.01); heart rate and mean arterial pressure were not significantly altered (P > 0.05). The lipid peroxidation products MDA and conjugated diene in myocardia were increased 22% (P < 0.01), 68% (P < 0.05) (P < 0.05), respectively. The plasma activity of CPK and LDH was greatly increased by 4.5-and 3.1-fold (P < 0.01), respectively. ACEI and spironolactone obviously relieved degree of calcium-overload and improved cardiac function and myocardial injury(P < 0.01). Calcium content in myocardia and artery was lower 44%, 39% and 57%, 34%. Lower MDA by 20%, 30%, lower conjugated diene by 44%, 35% than calcium-overload group. The plasma activity of CPK and LDH were obviously decreased 28%, 34% and 20%, 27%, compared with calcium-overload group.

CONCLUSIONCalcium-overload could lead to cardiac function descending and myocardial injury in calcium-overload rats by VDN. ACEI and spironolactone could reduce calcium-overload in myocardial and ameliorate cardiac function and decrease myocardial injury.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Calcium ; adverse effects ; Creatine Kinase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lipid Peroxidation ; Male ; Malondialdehyde ; analysis ; Mineralocorticoid Receptor Antagonists ; Myocardium ; metabolism ; Nicotine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spironolactone ; pharmacology ; Vitamin D ; pharmacology

OBJECTIVETo investigate the relationship between the chronotropic incompetence and angiographic severity in patients with coronary artery disease (CAD).

METHODSCoronary angiography was performed in 130 patients suspected for CAD and angiographic severity of coronary artery was quantitated by Duke score and Gensini score. Patients were divided to 4 groups: non-CAD group (39 patients), CAD group with one coronary artery involved (CHD1 group, 30 patients), CHD group with two coronary arteries involved (CHD2 group, 31 patients) and CAD group with three coronary arteries involved (CHD3 group, 30 patients). One month before coronary angiography, symptom-limited exercise treadmill tests were made and the ratio of heart rate reserve (HRR) and the percent maximal age-predicted heart rate achieved (rHR) were measured.

RESULTSrHR and HRR were significantly lower in CHD2 group (rHR 0.79+/-0.08, HRR 0.63+/-0.11) and CHD3 (rHR 0.78+/-0.07, HRR 0.59+/-0.12) than that in non-CHD group (rHR 0.89+/-0.06, HRR 0.80+/-0.10) and CHD1 group (rHR 0.86+/-0.08, HRR 0.74+/-0.15, all P<0.05). rHR and HRR also significantly correlated with Duke score (r=-0.554, -0.578, all P<0.01) and Gennisi score (r=-0.453, -0.467, all P<0.01). CHD incidence rate was 75% in patients with positive rHR (or HRR) but without ST lowering during exercise.

CONCLUSIONChronotropic incompetence are negatively related to angiographic coronary severities and thus predict angiographic coronary severities. There is a high CAD incidence in patients with positive rHR (or HRR) but no ST lowering during symptom-limited exercise treadmill tests.

Aged ; Coronary Angiography ; Coronary Artery Disease ; diagnosis ; diagnostic imaging ; Exercise Test ; Female ; Heart Rate ; Humans ; Male ; Middle Aged ; Severity of Illness Index