OBJECTIVETo establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field.

METHODSThree 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining.

RESULTSCompared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01).

CONCLUSION4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.

Alkaline Phosphatase ; metabolism ; Animals ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Chloral Hydrate ; pharmacology ; Cilia ; drug effects ; enzymology ; physiology ; Male ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis.

METHODSThirty-six female one-month-old SD rats of (125 +/- 3) g body weight were randomly divided into three groups, 12 rats in each group, one group was orally administered osthole at 9 mg x kg(-1) d(-1), one group was given genistein at 10 mg x kg(-1) d(-1) and another was given equal quantity of distilled water as the control. The body weight was monitored weekly and the bone mineral density (BMD) of total body was measured every month. All rats were sacrificed after three months, the femoral bone mineral density, the serum levels of osteocalcin (OC) and anti-tartaric acid phosphatase 5b (TRACP 5b) were measured by Elisa. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine.

RESULTSNo significant differences were observed between O-treated or GEN group and the control for the food-intake and body weight during three months. However, the rats treated with OST had significant higher BMD for both total body and femur than the control and GEN group. The O-treated rats also had higher level of serum OC and lower level of TRACP 5b. Besides, they owned bigger bone volume/tissue volume, trabecular thickness, trabecular number but smaller trabecular spacing. In the three point bending tests of femurs,they were found to have larger maximum load, the young's modulus and structural model index (SMI).

CONCLUSIONOrally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.

Acid Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Bone Density ; drug effects ; Coumarins ; pharmacology ; Female ; Femur ; diagnostic imaging ; drug effects ; pathology ; Genistein ; pharmacology ; Isoenzymes ; blood ; Osteocalcin ; blood ; Radiography ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

OBJECTIVETo observe effects of Ligustrazine on serum S100p protein and neuron-specific enolase (NSE) in elderly patients undergoing orthopedics operations.

METHODSTotally 60 patients undergoing selective total hip replacement, 65-80 years old, who were in line with American Society of Anesthesiologists (ASA) grade I or II, were randomly assigned to the Ligustrazine group (Group L) and the normal saline control group (Group S). The right internal jugular vein catheters were placedcephalad and ensured theirs tips in jugular venous bulbs after anesthesia induction and tracheal intubation. Patients in Group L received 2 mg/kg Ligustrazine Injection (40 drops within one minute) and those inGroup S received equal volume of normal saline via central veins before operations. Other medicines were the same for all patients during and after operation. Five millimeter blood sample was collected frominternal jugular venous bulbs before operation (T0), 24 h (T1), 72 h (T2), 168 h (7th day, T3) after operation. Serum was collected after centrifuge. S100β protein and NSE concentration were analyzed usingELISA. Mini-mental state examinations (MMSE) were scored by the same doctor at T0, T1, T2, and T3,respectively.

RESULTSThere was no statistical difference in MMSE scores, serum S1000 protein, or NSE at TO (P > 0.05). Compared with TO, S100 P protein and NSE concentration increased and MMSE scores decreased at T1, T2, and T3 in the two groups. All indices except S100P protein and NSE at T3 were statistically different between Group L and Group S (P < 0.05).

CONCLUSIONSerum S100P protein and NSE could be changed by pre-operation injecting Ligustrazine at certain dose in elderly patients undergoing orthopedics operations.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Humans ; Phosphopyruvate Hydratase ; blood ; Pyrazines ; therapeutic use ; S100 Calcium Binding Protein beta Subunit ; blood

This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.
Alkaline Phosphatase ; metabolism ; Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Calcium ; metabolism ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Enzyme Activation ; drug effects ; Femur ; metabolism ; Gene Expression Regulation ; Genistein ; pharmacology ; Osteoprotegerin ; genetics ; metabolism ; Phytoestrogens ; pharmacology ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the effects of static magnetic fields (SMFs) with different exposure time on the maturation of rat osteoblasts in vitro and the expression of the estrogen receptor (ER) gene.

METHODSThe calvarial osteoblasts were isolated from newborn rats by enzyme digestion and randomly divided into 9 groups after one passage based on the exposure time of the SMFs[0 (control), 0.5 h, 1.0 h, 1.5 h, 2.0 h, 2.5 h, 3.0 h, 3.5 h, and 4.0 h]. The intensity was 3.9 mT in all SMFs. Those without SMFs exposure were used as the controls. The oeteoblasts were observed under the contrast phase microscope on a daily basis. After 48 h, cell proliferation was assayed by MTT method. The osteocalcin contents were measured after exposure to SMFs for 3 d, 6 d, 9 d, and 12 d. ERΑ and ERΒ mRNA expressions were measured by real-time PCR after SMFs treatment for 0 h, 24 h, 48 h, and 72 h.

RESULTSCompared with the controls, the cell proliferation was significantly enhanced in the 2.0-h, 2.5-h, and 3.0-h groups (P<0.05). After SMFs treatment for 6 d, 9 d and 12 d, the 2.5-h group had significantly higher osteocalcin content than the control group did (P<0.05). After SMFs treatment for 0 h and 72 h, elevated ERΑ mRNA expression and reduced ERΒ mRNA expression were observed.

CONCLUSIONExposure to SMFs, regardless of exposure time, is associated with enhanced cell proliferation, increased osteocalcin contents, and altered ERΑ and ERΒ mRNA expressions in opposite directions.

Animals ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Magnetic Fields ; Osteoblasts ; cytology ; metabolism ; Rats ; Receptors, Estrogen ; genetics ; metabolism

OBJECTIVETo investigated the effect of 50-Hz 3.6-mT sinusoidal electromagnetic fields (SEMFs) on the proliferation and differentiation of osteoblasts in vitro.

METHODSThe newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 6 groups after one passage. The treatment groups under 50-Hz 3.6-mT SEMFs and controls without SEMFs treatment. The cells were exposed in the SEMFs for 0.5 h, 1.0 h, 1.5 h, 2.0 h, and 2.5 h. They were observed under the contrast phase microscope each day. The calcified nodules were stained by alizarin red. The SEMFs were arranged in spiral appearance after 3 to 5 days.

RESULTSThe SEMFs showed characteristic distribution 3 to 5 days after SEMFs treatment. On the 9(th) day after treatment, the activity of alkaline phosphatase (ALP) significantly increased in the 0.5-h group, whereas the ALP histochemical straining results and the area of calcified nodules were consistent with ALP activity. In the 48-h and 96-h groups, the genetic expression levels of osteoprotegerin and collagen-1 were significantly higher than that in the control group; particularly, the mRNA expression increased in the 0.5-h group.

CONCLUSIONThe SEMFs at 50-Hz 3.6-mT could suppress the proliferation of osteoblasts maturation but stimulate the differentiation and maturation of osteoblasts in vitro.

Animals ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Male ; Osteoblasts ; cytology ; radiation effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of exposure to static magnetic fields (SMFs) of 3.9 mT on proliferation and differentiation of osteoblasts in vitro.

METHODSThe newborn rat calvarial osteoblasts were isolated by enzyme digestion and randomly divided into 9 groups after one passage. The intensity of the SMFs was 3.9 mT. The cells were exposed in the SMFs for 0 (control group), 0.5, 1.0, 1.5, 2.0, 2.5, 3, 3.5 and 4.0 h groups respectively. They were observed under the contrast phase microscope each day. After 48 h, cell proliferation was assayed by MTT method. The alkaline phosphatase (Alkaline Phosphatase, ALP) activities and calcium content were measured after 3, 6, 9, and 12 days exposed with SMFs. The ALP positive colonies were histochemically stained after 8 days and the calcified nodules were stained by Alizarin Bordeaux after 10 days; BMP-2, Runx-2 and Opg mRNA expression were measured after SMFs treatment in 0, 24, 48 and 72 h.

RESULTSContrast with control group, all SMFs groups enhanced cell proliferation (P < 0.01 or P < 0.05), and they promoted maturation and mineralization of the osteoblasts. The results showed that SMFs improved the ALP activity, promoted calcium content, boost BMP-2, Runx -2 and Opg mRNA expression.

CONCLUSIONThe cells exposed to the SMFs of 3.9 mT at 2.5 h apparently promote proliferation and differentiation of osteoblasts in vitro.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; Calcium ; metabolism ; Cell Differentiation ; radiation effects ; Cell Proliferation ; radiation effects ; Core Binding Factor Alpha 1 Subunit ; genetics ; Magnetic Fields ; Osteoblasts ; physiology ; radiation effects ; Osteoprotegerin ; genetics ; Rats ; Rats, Sprague-Dawley ; Time Factors

Objective To analyze the timeliness of detection and reporting on public health emergency events, and to explore the effective strategies for improving the relative capacity on those issues. Methods We conducted a retrospective survey on 3275 emergency events reported through Public Health Emergency Events Surveillance System from 2005 to the first half of 2006. Developed by county Centers for Disease Control and Prevention, a uniformed self-administrated questionnaire was used to collect data, which would include information on the detection, reporting of the events. Results For communicable diseases events, the median of time interval between the occurrence of first case and the detection of event was 6 days (P25=2, P75=13). For food poisoning events and clusters of disease with unknown origin,the medians were 3 hours (P25=1, P75=16) and 1 days (P25=0,P75=5). 71.54% of the events were reported by the discoverers within 2 hours after the detection. Conclusion In general, the ranges of time intervals between the occurrence, detection or reporting of the events were different, according to the categories of events. The timeliness of detection and reporting of events could have been improved dramatically if the definition of events, according to their characteristics, had been more reasonable and accessible, as well as the improvement of training program for healthcare staff and teachers.

OBJECTIVETo investigate the effects of different-intensity sinusoidal electromagnetic fields (SEMFs) on bone mineral density (BMD) and histomorphometry in SD rats.

METHODSThirty female SD rats were randomly divided into three groups: group A (a control group), group B (0.1 mT group) and group C (0.6 mT group). The rats in group B and C were exposed to 50 Hz SEMFs 3 hours each day. However,the magnetic intensity was different between group B and group C:0.1 mT for group B and 0.6 mT for group C. After 8 weeks, all the animals were killed. Changes of BMD and histomorphometric properties were observed.

RESULTSCompared with group A, the BMD of whole body, femur and vertebrae of rats in group B increased significantly; the area percentage, number and width of bone trabeculae in vertebrae and femur of rats in group B were larger than those of group A; but the resolution of bone trabeculae of rats in group B was lower than that of group A. The trabecular number in group C rats were significantly decreased, compared with that in group A rats. The outcome of double fluorescence labeling in group B was found to be significantly different with that in group A. But the difference between rats in group A and C was not significant.

CONCLUSIONThis study demonstrates that 50 Hz 0.1 mT SEMFs can increase BMD, improve bone tissue microstructure and, promote bone formation.

Animals ; Bone Density ; radiation effects ; Electromagnetic Fields ; Female ; Lumbar Vertebrae ; pathology ; radiation effects ; Osteogenesis ; radiation effects ; Rats ; Rats, Sprague-Dawley ; Tibia ; pathology ; radiation effects

OBJECTIVETo verify the clinical efficacy of acupuncture on motor dysfunction in ischemic stroke of subacute stage.

METHODSThe multi-central randomized controlled trial was adopted. One hundred and twenty-six cases of ischemic stroke of subacute stage were randomized into an acupuncture group (61 cases) and a conventional treatment group (65 cases). The basic treatment of western internal medicine and rehabilitation training were applied to the patients of the two groups. In the acupuncture group, acupuncture was supplemented at the body points located on the extensor of the upper limbs and the flexor of the lower limbs. In combination, scalp acupuncture was applied to NS5, MS6 and MS6 on the affected side. The treatment was given 5 times a week and totally 8 weeks were required. The follow-up observation lasted for 3 months. The scores in Fugl-Meyer scale and NIHSS scale and Barthel index were compared between the two groups before treatment, in 4 and 8 weeks of treatment and the 3-month follow-up observation after treatment separately.

RESULTSIn 4 and 8 weeks of treatment and the follow-up observation, Fugl-Meyer scale score was improved obviously in the patients of the two groups (all P<0. 01). In 8 weeks of treatment and the follow-up observation, Fugl-Meyer scale score in the acupuncture groupwas im proved much apparently as compared with that in the conventional treatment group [68. 0 (43. 0,86. 5) vs 52. 5 (30.3, 77.0), 77.0 (49.5, 89.0) vs 63. 0 (33.0, 84.0), both P<0. 05]. Except that NIHSS scale score was not reduced apparently in 4 weeks of treatment in the conventional treatment group (P>0.05), the results of NIHSS scale at the other time points were all decreased obviously as compared with those before treatment in the patients of the two groups (all P<0. 01). In 8 weeks of treatment and the follow-up observation, the results in the acupuncture group were reduced much apparently as compared with those in the conventional treatment group [5. 0 (3.0,8.0) vs 7. 0 (3.0,13.8), 4. 0 (1.5,7.0) vs 6.0 (2.0,11.7) ,both P<0. 05]. In 8 weeks of treatment and the follow-up observation, Barthel index was improved obviously as compared with that before treatment in the patients of the two groups (all P<0. 05). The improvement in the acupuncture group was much more significant as compared with the conventional treatment group [75. 0 (60. 0,87. 5) vs 65. O (36. 3, 87. 5), P<0. 051.

CONCLUSIONBased on the conventional treatment, Acupuncture achieves the satisfactory clinical efficacy on motor dysfunction in ischemic stroke of subacute stage.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Motor Activity ; Stroke ; physiopathology ; therapy ; Treatment Outcome