ABSTRACT
Objective: To investigate the effect of an aqueous extract of Protaetia brevitarsis (AEPB) on osteogenesis using preosteoblast MC3T3-E1 cells and zebrafish larvae. Methods: Flow cytometric analysis was used to measure the cytotoxicy. Alkaline phosphatase activity was detetmined using p-nitrophenyl phosphate as a substrate. Calcium deposition was detected using alizarin red staining along with osteogenic marker expression in preosteoblast MC3T3E1 cells. In addition, vertebral formation in zebrafish larvae was detected using calcein staining and osteogenic gene expression. Results: AEPB highly promoted the expression of osteogenic markers including runt-related transcription factor 2, osterix, and alkaline phosphatase, along with elevated levels of mineralization in MC3T3-E1 cells. Moreover, AEPB accelerated vertebral formation in zebrafish larvae accompanied by upregulated expression of osteogenic genes. FH535, an inhibitor of Wnt/β-catenin, suppressed AEPB-induced osteogenic gene expression and vertebral formation, indicating that AEPB stimulates osteogenesis by activating the Wnt/β-catenin signaling pathway. Conclusions: AEPB stimulates osteoblast differentiation and bone formation by activating β-catenin. Therefore, AEPB is a promising material that induces osteogenesis, and is useful for the treatment of bone resorption diseases.
ABSTRACT
Objective: To investigate the effect of an aqueous extract of Protaetia brevitarsis (AEPB) on osteogenesis using preosteoblast MC3T3-E1 cells and zebrafish larvae. Methods: Flow cytometric analysis was used to measure the cytotoxicy. Alkaline phosphatase activity was detetmined using p-nitrophenyl phosphate as a substrate. Calcium deposition was detected using alizarin red staining along with osteogenic marker expression in preosteoblast MC3T3E1 cells. In addition, vertebral formation in zebrafish larvae was detected using calcein staining and osteogenic gene expression. Results: AEPB highly promoted the expression of osteogenic markers including runt-related transcription factor 2, osterix, and alkaline phosphatase, along with elevated levels of mineralization in MC3T3-E1 cells. Moreover, AEPB accelerated vertebral formation in zebrafish larvae accompanied by upregulated expression of osteogenic genes. FH535, an inhibitor of Wnt/β-catenin, suppressed AEPB-induced osteogenic gene expression and vertebral formation, indicating that AEPB stimulates osteogenesis by activating the Wnt/β-catenin signaling pathway. Conclusions: AEPB stimulates osteoblast differentiation and bone formation by activating β-catenin. Therefore, AEPB is a promising material that induces osteogenesis, and is useful for the treatment of bone resorption diseases.
ABSTRACT
OBJECTIVE@#To evaluate whether the methanol extract of Codium fragile (MECF) regulates tumor necrosis factor-α (TNF-α)-induced invasion of human breast cancer MDA-MB-231 cells by suppressing matrix metalloproteinase-9 (MMP-9).@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis were performed to analyze the expression of MMP-9 and nuclear factor-κB (NF-κB) subunits, p65 and p50, and IκB in MDA-MB-231 cells. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for cell viability. MMP-9 activity and invasion were measured by gelatin zymography and a matrigel invasion assay, respectively. NF-κB activity was measured by an electrophoretic mobility shift assay and luciferase activity.@*RESULTS@#MECF had no effect on cell viability up to a concentration of 100 μg/mL in human breast cancer MDA-MB-231 cells regardless of the presence of TNF-α. MDA-MB-231 cells that were stimulated with TNF-α showed a marked increase of invasion compared to the untreated control, whereas pretreatment with MECF downregulated the TNF-α-induced invasion of MDA-MB-231 cells. Additionally, zymography, western blot analysis, and RT-PCR confirmed that MECF decreased TNF-α-induced MMP-9 expression and activity which is a key regulator for cancer invasion. According to an electrophoretic morbidity shift assay, pretreatment with MECF in MDA-MB-231 cells significantly decreased the TNF-α-induced DNA-binding activity of NF-κB, which is an important transcription factor for regulating cancer invasion-related genes such as MMP-9. Furthermore, treatment with MECF sustained the expression of p65 and p50 in response to TNF-α in the cytosolic compartment. The luciferase assay demonstrated that MECF attenuated TNF-α-induced NF-κB luciferase activity.@*CONCLUSION@#MECF exhibited its anti-invasive capability by downregulating TNF-α-induced MMP-9 expression, resulting from the suppression of NF-κB activity in the human breast cancer cell line MDA-MB-231.
ABSTRACT
Objective To evaluate whether the methanol extract of Codium fragile (MECF) regulates tumor necrosis factor-α (TNF-α)-induced invasion of human breast cancer MDA-MB-231 cells by suppressing matrix metalloproteinase-9 (MMP-9). Methods Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis were performed to analyze the expression of MMP-9 and nuclear factor-κB (NF-κB) subunits, p65 and p50, and IκB in MDA-MB-231 cells. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for cell viability. MMP-9 activity and invasion were measured by gelatin zymography and a matrigel invasion assay, respectively. NF-κB activity was measured by an electrophoretic mobility shift assay and luciferase activity. Results MECF had no effect on cell viability up to a concentration of 100 μg/mL in human breast cancer MDA-MB-231 cells regardless of the presence of TNF-α. MDA-MB-231 cells that were stimulated with TNF-α showed a marked increase of invasion compared to the untreated control, whereas pretreatment with MECF downregulated the TNF-α-induced invasion of MDA-MB-231 cells. Additionally, zymography, western blot analysis, and RT-PCR confirmed that MECF decreased TNF-α-induced MMP-9 expression and activity which is a key regulator for cancer invasion. According to an electrophoretic morbidity shift assay, pretreatment with MECF in MDA-MB-231 cells significantly decreased the TNF-α-induced DNA-binding activity of NF-κB, which is an important transcription factor for regulating cancer invasion-related genes such as MMP-9. Furthermore, treatment with MECF sustained the expression of p65 and p50 in response to TNF-α in the cytosolic compartment. The luciferase assay demonstrated that MECF attenuated TNF-α-induced NF-κB luciferase activity. Conclusion MECF exhibited its anti-invasive capability by downregulating TNF-α-induced MMP-9 expression, resulting from the suppression of NF-κB activity in the human breast cancer cell line MDA-MB-231.
ABSTRACT
The JNK inhibitor SP600125 strongly inhibits cell proliferation in many human cancer cells by blocking cell-cycle progression and inducing apoptosis. Despite extensive study, the mechanism by which SP600125 inhibits mitosis-related effects in human leukemia cells remains unclear. We investigated the effects of SP600125 on the inhibition of cell proliferation and the cell cycle, and on microtubule dynamics in vivo and in vitro. Treatment of synchronized leukemia cells with varying concentrations of SP600125 results in significant G2/M cell cycle arrest with elevated p21 levels, phosphorylation of histone H3 within 24 h, and endoreduplication with elevated Cdk2 protein levels after 48 h. SP600125 also induces significant abnormal microtubule dynamics in vivo. High concentrations of SP600125 (200 microMeter) were required to disorganize microtubule polymerization in vitro. Additionally, SP600125-induced delayed apoptosis and cell death was accompanied by significant poly ADP-ribose polymerase (PARP) cleavage and caspase-3 activity in the late phase (at 72 h). Endoreduplication showed a greater increase in ectopic Bcl-2-expressing U937 cells at 72 h than in wild-type U937 cells without delayed apoptosis. These results indicate that Bcl-2 suppresses apoptosis and SP600125-induced G2/M arrest and endoreduplication. Therefore, we suggest that SP600125 induces mitotic arrest by inducing abnormal spindle microtubule dynamics.
Subject(s)
Humans , Anthracenes/therapeutic use , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Cell Cycle/drug effects , Enzyme Inhibitors/therapeutic use , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , Leukemia/drug therapy , Tubulin/metabolismABSTRACT
Subject(s)
Humans , Anesthesia, Local , Conscious Sedation , Joints , Mouth , Recurrence , Splints , Therapeutic IrrigationABSTRACT
Subject(s)
Humans , Ankylosis , Arthroplasty , Chin , Chronic Pain , Facial Asymmetry , Joints , Molar , Mouth , Oral Hygiene , Temporomandibular JointABSTRACT
From January, 1992 to June, 1996, )7 patients with flail chest were treated at Sonnchunhyang university hospital. 15 patients were managed by internal fixation of fractured ribs, whereas the remaining 22 patients were managed by endotracheal intubation and intermittent positive-pressure ventilation alone. There were no difference between two groups in age, sex, the severity of injury to the chest wall and the nature of associated injuries. Average dur'Btion of assisted ventilation was 5.7 +/-1.7 days in the patients treated by internal fixation versus 8.7 +/-3.3 days In the patients treated by continuous me hanical ventilation. Average stay in the intensive care unit was 8.3 +/-3.9 days for the patients treated by internal fixation, whereas it was 13.2+/-4.1 days in the group treated by continuous mechanical ventilation alone. In the group treated by internal fixation, complications were 3 atelectases(20.0%), 1 pneumonia(6.7%), 2 operative wound problems(12.3%) and 1 barotrauma(6.7%). In the other group, 7 atelectases(31.8%), 4 pneumonitis(18.2%), 2 empyemas(9.1%) and 3 barotraumas(1).6%). The mortality rate was 13.3%(2/15) in the surgically treated patients, whereas it was 22.7%(5122) in the other group. The treatment of flail chest by internal fixation resulted in speedy recovery, decreased complications and mortalities, and better ultimatc cosmetic and functional results.
Subject(s)
Humans , Flail Chest , Intensive Care Units , Intermittent Positive-Pressure Ventilation , Intubation, Intratracheal , Mortality , Respiration, Artificial , Ribs , Thoracic Wall , Ventilation , Wounds and InjuriesABSTRACT
Hypothermia during lung preservation decreases metabolic processes. After the rabbit lung was flushed with modified Euro-Collins solution, heart-lung block was harvested and the left lung was assessed after ligation of the right pulmonary artery and right main-stem bronchus. Heart-lung block was immersed in the same solution for 6 hours. The modified Euro-Collins solution and storage temperature of group 1( 10 cases ) was 4 degrees C, group 2( 10 cases ) was 10 degrees C. On completion of the storage period, the left lung was ventilated and reperfused with blood which used a cross-circulating paracorporeal rabbit as a " biologic deoxygenator " for 60 minutes. Pulmonary artery pressure, airway pressure, difference in oxygen tension between inflow and outflow perfusate and degree of pulmonary edema were assessed at 10-minute intervals while the left lung was ventilated at 0.8 of the inspired oxygen fraction. The mean pulmonary venous oxygen tensions at 10 and 60 minutes after reperfusion were 209.52 +/- 42.46 and 103.48 +/- 15.96 mmHg in group I versus 247.78 +/- 36.19 and 147.91 +/- 11.07 mmHg in group II( p = 0.049, <0.0001 ). The mean alveolar-arterial oxygen differences at 20 and 60 minutes after reperfusion were 357.95 +/- 12.84 and 437.31 +/- 14.26 mmHg in group I versus 310.88 +/- 33.47 and 390.93 +/- 15.86 mmHg in group II( p = 0.0092, <0.0001 ). The mean pulmonary arterial pressures at 10 and 60 minutes after reperfusion were 40.56 +/- 18.66 and 87.22 +/- 17.22 mmHg in group I versus 31.22 +/- 6.84 and 65.78 +/- 11.02 mmHg in group II( p = 0.048, 0.0062 ). The mean pulmonary vascular resistances at 10 and 60 minutes after reperfusion were 2.69 +/- 0.85 and 4.36 +/- 0.86 mmHg/ml/min in group I versus 1.99 +/- 0.39 and 3.29 +/- 0.55 mmHg/ml/min in group II( p = 0.0323, 0.0062 ). There were no difference between groups in peak airway pressure, lung compliance and degree of pulmonary edema. In conclusion that preservation of lung at 10 degrees C was superior to preservation at 4 degrees C.
Subject(s)
Arterial Pressure , Bronchi , Hypothermia , Ligation , Lung Compliance , Lung Transplantation , Lung , Metabolism , Organ Preservation , Oxygen , Pulmonary Artery , Pulmonary Edema , ReperfusionABSTRACT
No abstract available.
