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The wrap around the toe flap from the great toe is considered to be a good reconstructive proce-dure for degloved injuries of the thumb. In this study, we used prefabricated flaps of the medial plantar skin to cover a degloved injury of the thumb of a patient unsuitable for application of this method and obtained satisfactory clinical results. [ABSTRACT FROM AUTHOR]

Objective: To 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., Methods: Three 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., Results: Compared 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)., Conclusion: 4mM 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.

Objectives: Icariin, a prenylated flavonol glycoside isolated from traditional Chinese medicinal herb of the genus Epimedium, has been demonstrated to be a potential alternative therapy for osteoporosis, and its action mechanism so far has been mainly attributed to its phytoestrogenic property. As blood supply to bone is considerably reduced with ageing and by the menopause, we hypothesized that icariin treatment would reduce bone loss by preventing ischaemia-induced hypoxic damages to bone., Materials and Methods: To investigate effects of icariin treatment on cultured rat calvarial osteoblasts exposed to hypoxic conditions (2% oxygen)., Results: Compared to normoxic control, cell viability decreased with time to 50% by 48 h in the hypoxic group, and icariin attenuated the reduction, dose dependently, with 10(-6) and 10(-5)  m concentrations showing significant protective effects. Icariin also inhibited increase of lactate dehydrogenase activity in culture media. Measurements on oxidative stress, cell cycling and cell survival indicated that icariin protected osteoblasts by reducing production of reactive oxygen species and malondialdehyde, increasing superoxide dismutase activity, arresting the cell cycle and inhibiting apoptosis. Icariin also preserved osteogenic differentiation potential of the hypoxic cells in a dose-dependent manner, compared to the hypoxia alone group, as revealed by increased levels of RUNX-2, OSX and BMP-2 gene expression, alkaline phosphatase activity, and formation of mineralized nodules., Conclusions: Our results demonstrated that icariin attenuated oxidative stress and apoptosis and preserved viability and osteogenic potential of osteoblasts exposed to hypoxia in vitro, and suggested that its anti-osteoporotic effect may be attributed to its anti-hypoxic activity and phytoestrogenic properties., (© 2014 John Wiley & Sons Ltd.)

Objective: To investigate the effects of different-intensity sinusoidal electromagnetic fields (SEMFs) on bone mineral density (BMD) and histomorphometry in SD rats., Methods: Thirty 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., Results: Compared 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., Conclusion: This study demonstrates that 50 Hz 0.1 mT SEMFs can increase BMD, improve bone tissue microstructure and, promote bone formation.

Icariin, a prenylated flavonol glycoside isolated from Epimedii herba, has been found to be a potent stimulator of osteogenic differentiation and has potential application in preventing bone loss. However, the signaling pathway underlying its osteogenic effect remains unclear. We hypothesized that the osteogenic activity of icariin is related to the nitric oxide (NO) signal pathway and PI3K/AKT pathway in its upstream. Rat bone marrow stromal cells (rBMSCs) were cultured in osteogenic medium and treated with icariin or together with L-NAME, ODQ, PDE5, and/or LY294002 (the inhibitor of NOS, sGC, cGMP, and PI3K respectively), and effects were examined on the expression of signal messengers (NOS, NO, sGC, cGMP, PKG and PI3K) and the levels of osteogenic markers (alkaline phosphatase or ALP, osteocalcin and calcified nodules). It was found that icariin dose-dependently increased ALP activity, and treatment at the optimal concentration (10(-5)M) increased NOS activity, iNOS and eNOS expression, NO production, sGC and cGMP contents and PKG expression besides the phosphorylation of AKT. The addition of L-NAME, ODQ and PDE5 significantly inhibited the icariin effects on above markers respectively. The addition of LY294002 decreased the p-AKT level, NOS activity, eNOS expression and NO production significantly, but had no significant effect on iNOS expression. The addition of any of the four inhibitors also abolished the osteogenic effect of icariin on rBMSCs as indicated by ALP activity, osteocalcin synthesis, calcium deposition and the number and areas of calcified nodules. These results suggest that the osteogenic effect of icariin involves the PI3K-AKT-eNOS-NO-cGMP-PKG signal pathway. Furthermore, dosage response studies showed that icariin at 10(-6)M (a physiologically achievable concentration in vivo) also activated this signal pathway., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: To compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis., Methods: Thirty-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., Results: No 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)., Conclusion: Orally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.

Objective: Through researching the relationship among osteoporosis and inflammatory reaction besides angiogenesis, to compare pharmacological differences between icariin and genistein to inhibit bone loss., Methods: 6 months old female SD rats were randomly divided into SHAM group, model group, ICA group, GEN group and E group. The bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and tibia microarchitecture were analyzed., Results: Compared with SHAM group, model group of body weight, uterine weight, bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and lumbar and tibia microarchitecture were significantly changed (P < 0.05). Compared with model group, ICA group of body weight, bone mineral density of total and femur, serum TRACP 5b and femural biomechanics were significantly changed (P < 0.05). GEN group of bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and lumbar and tibia microarchitecture were significantly changed (P < 0.05)., Conclusion: Icariin inhibits bone loss on model rat through suppressing bone resorption. Genistein prevents bone loss on model rat by the pathway of inhibiting inflammatory reaction, activating angiogenesis, enhancing bone formation and inhibiting bone resorption. Moreover, pharmacological activity of genistein is more potential than icariin.

Noninvasive electromagnetic fields (EMFs) have been known to be able to improve bone health; however, their optimal application parameters and action mechanisms remain unclear. This study compared the effects of different forms of EMFs (sinusoidal, triangular, square, and serrated, all set at 50 Hz frequency and 1.8 mT intensity) on proliferation, differentiation and mineralization of rat calvarial osteoblasts. Square EMFs stimulated osteoblast proliferation but sinusoidal EMFs inhibited it. Sinusoidal and triangular EMFs produced significantly greater alkaline phosphatase (ALP) activity, ALP staining areas, calcium deposition, mineralized nodule areas, and mRNA expression of Runx-2, osteoprotegerin and insulin-like growth factor-I than square and serrated EMFs (P < 0.01). Triangular EMFs had a greater effect than sinusoidal EMFs on every indices except for Runx-2 mRNA expression (P < 0.05). These results indicated that while square EMFs promoted proliferation and had no effect on the differentiation of osteoblasts, sinusoidal EMFs inhibited proliferation but enhanced osteogenic differentiation. Triangular EMFs did not affect cell proliferation but induced the strongest osteogenic activity among the four waveforms of EMFs. Thus, the effects of EMFs on proliferation and differentiation of osteoblasts in vitro were dependent on their waveforms., (Copyright © 2013 Wiley Periodicals, Inc.)

An effective method for preventing bone loss is by promoting osteoblast differentiation and bone formation. While dexamethasone has been routinely used as a classical inducer for osteoblast differentiation, limitations have been observed with its usage, including its varied effects on expression of osteoblast genes in different species and its potentials in suppressing osteoblastic differentiation and mineralization. In this study, we assessed the ability of flavonoid icariin in enhancing differentiation and mineralization of cultured rat primary osteoblasts in the absence of dexamethasone. It was found that, compared to the non-stimulated control, icariin at 10(-5) M produced a higher alkaline phosphatase activity, more and larger areas of alkaline phosphatase-positive colonies (CFU-FALP) and mineralized nodules, more osteocalcin secretion and calcium deposition, higher levels of mRNA expression of alkaline phosphatase, osteoblastic transcription factors osterix and runt-related transcription factor 2, and collagen 1α, higher levels of protein expression of collagen 1α, alkaline phosphatese, osterix, and runt-related transcription factor 2. In addition, icariin at 10(-5) M was always more potent than dexamethasone at its optimal concentration of 10(-8) M on the above osteoblast differentiation and mineralization markers. Taken together, our studies demonstrated that icariin has a pronounced ability in promoting osteoblast differentiation in vitro in the absence of dexamethasone., (Georg Thieme Verlag KG Stuttgart · New York.)

Objective: To compare the effects of icariin (ICA) and genistein (GEN) on rats bone peak mass and thus screen for a drug that can more effectively prevent osteoporosis., Methods: Totally 36 one-month SD rats were randomly divided into three groups: ICA group [25 mg/(kg·d), intragastric administration], GEN group [10 mg/(kg·d), intragastric administration], and control group (fed with equal volume of distilled water). The body weight was monitored weekly and the bone mineral density of total body was measured monthly. All rats were sacrificed three months later. The femoral bone mineral density and the serum levels of osteocalcin and anti-tartaric acid phosphatase 5b, N-terminal propeptide of type 1 procollagen, and C-terminal propeptide of type 1collagen were measured. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine., Results: The body weight and organ index showed no significant difference among these three groups(P>0.05). No obvious pathological change was found. The bone mineral density was also not significantly different in the first and second months; however, in the third months, the ICA group had significant higher bone mineral density for both total body and femur than those in the control and GEN group (P<0.05). The same trends were found for both femur bone mineral density and whole-body bone mineral density (P<0.05). The ICA group also had significantly higher serum levels of osteocalcin (P<0.05) and lower level of anti-tartaric acid phosphatase 5b(P<0.05). Besides, rats in the ICA group had significantly larger bone volume/tissue volume, trabecular thickness, and trabecular number than the control group, whereas the trabecular spacing and model coefficients were signicantly lower(all P<0.05), which, however, were not significantly different between ICA group and GEN group (P>0.05). Femoral maximum load, Youg's modulus, and yield load were significantly higher in these two groups than in the control group (P<0.05), which, again, were not significantly different between ICA group and GEN group (P>0.05)., Conclusion: Orally administered ICA is more efficient than GEN in inhibiting resorption and promoting bone formation, and thus can dramatically improve the peak bone mineral density and bone quality.

Objective: To investigate the effect of osthole on bone metabolism in rat femoral tissues in vitro., Methods: The rat femoral tissues were isolated in vitro. The optimal concentrations of ostehole (1×10(-5) mol/L) and estradiol (1×10(-8) mol/L) (the positive control) were selected by alkaline phosphatase activity (ALP). The ALP and calcium levels were detected by commmerical regents, and the expressions of osteoprotegerin, receptor activator of nuclear factor-κB ligand, runx-related gene 2, and bone morphogenetic protein-2 mRNA were determined by real-time reverse transcription-polymerase chain reaction., Result: The osthole (1×10(-5) mol/L) significantly increased the activity of ALP, calcium level as well as the expressions of osteoprotegerin, receptor activator of nuclear factor-κB ligand, runx-related gene-2 and bone morphogenetic protein-2 mRNA in rat femoral tissues in vitro., Conclusion: Osthole can improve calcium level and ALP activity and regulate the bone metabolism-related genes in rat femoral tissues.

Objective: To investigate the molecular mechanisms of icariin (ICA) in regulating the bone formation of osteoblasts and the bone resorption of osteoclasts., Methods: Primary osteoblast cell cultures were obtained from newborn rat calvarial. Calcified nodules were stained by alizarin red. The mRNA levels of osterix (OSX), runt-related transcription factor 2 (Runx-2), alkaline phosphatase (ALP), Collagen1, osteoprotegerin (OPG), and receptor activator of nuclear factor-ΚB ligand (RANKL) were analyzed by quantitative real-time RT-PCR, the protein levels of OPG, RANKL, and Collagen1 were examined by Western blotting, and the intracellular Ca(2+) concentration of osteoblasts was measured on a flow cytometer using the Cellquest program., Results: Compared with control group, ICA markedly promoted bone formation by significant up-regulating the gene expressions of OSX, Runx-2,ALP, and Collagen1, the protein expression of Collagen1(all P<0.01), and the Ca(2+) concentration. Furthermore, ICA remarkably inhibited bone resorption by significant up-regulating the mRNA and protein expressions of OPG as well as the OPG/RANKL ratio., Conclusions: ICA could promote bone formation of osteoblasts through inducting the gene expressions of OSX,Runx-2, ALP and Collagen1, and the protein expressions of Collagen1, and by increasing the Ca (2+) concentration. Moreover, ICA could inhibit bone resorption of osteoclasts through regulating OPG/RANKL signal pathway.

Objective: To analyze the reason of postoperative recurrence of spinal tuberculosis and observe the clinical outcome of these patients in reoperation., Methods: From January 2002 to May 2010,27 patients with postoperative recrudescent spinal tuberculosis were treated. There were 15 males and 12 females with an average age of 36.5 years old (ranged, 21 to 65). The risk factors and effect strength associated with postoperative recrudescent spinal tuberculosis were compared by Logistic regression analysis. Individual operation was performed according to the major reason. Re-operative methods including debridment in 5 cases, debridment and sinuses resection in 7 cases, one stage debridement and bone grafting via anterior approach and internal fixation via posterior approach in 8 cases, one stage bone grafting and internal fixation via posterior approach combined with CT-guided percutaneous catheter drainage and local chemotherapy in 2 cases, CT-guided percutaneous catheter drainage and local chemotherapy in 5 cases. Antituberculosis drugs were regularly used in all patients after operation. The ESR, X-ray and 3D-CT were regularly performed to estimate the progress of tuberculosis and condition of bony fusion., Results: The risk factors associated with postoperative recrudescent spinal tuberculosis were complicated, including no regularly used antituberculosis drugs before and after operation, no early diagnosis and treatment of the postoperative fluidify, malnutrition, no thoroughly debridement during operation and poor spinal stability after operation, according to effect strength to arrange. There was no injury of blood vessel,spinal cord or ureter during reoperation. The follow-up period was from 12 to 36 months with an average of 24 months. Tuberculosis symptoms disappeared after reoperation and no complications such as tuberculosis recurrence, infection of incision, sinuses formation and internal fixation failure were found in the patients. ESR recovered normal in follow-up and bone graft obtained fusion at 8 to 12 months after operation and internal fixation position was normal., Conclusion: The reoperative reasons of spinal tuberculosis are complicated and multifactorial. The diagnosis and treatment are difficult. It is important to analyze the recrudescent reasons thoroughly before operation,emphasize the application of regularly antituberculosis drugs and individual operation, meanwhile, reinforce nourishment and supportive treatment.

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.

Objective: To compare the effect of icariin and genistein in the osteogenic differentiation of rat bone marrow stromal cells (rBMSC)., Method: Rat marrow stromal cells were seperated in vitro, and the optimal concentration of genisten and icriin were screened. Genistein and icariin with the concentration of 1 x 10(-5) mol x L(-1) were adopted to intereven rBMSCs cultured in vitro. Alkaline phosphatase (ALP) was determined at 3, 6, 9, 12,15 d after intervention; calcified nodule was detected with alizarin red staining at 12 d; OXS, Runx-2, bone morphogenetic protein (BMP-2) and Collagen-I mRNA expression were observed with Real-time RT-PCR at 12, 24, 48, 72, 96 h., Result: Genistein and icariin with the concentration of 1 x 10(-5) mol x L(-1) could increase the activity of ALP and the content of Ca, regulate OXS, BMP-2, Runx-2 and Collagen-I mRNA expression., Conclusion: Icariin showed a stronger effect in improving the osteogenic differentiation of rat bone marrow stromal cells than genistein.

Objective: To compare the effects of 8-prenylnaringenin (PNG) and naringenin (NG) on the activity and apoptosis of osteoclasts cultured in vitro, in order to study physiological activity of 8-prenyl perssad., Method: Osteoclasts were separated from long-limb bones of newly born rabbits, cultured in alpha-MEM containing 10% FBS, and then added with PNG and NG with the concentration of 1 x 10(-5) mol x L(-1). They were stained with TRAP and determined for enzymatic activity with TRAP after 4 d, and analyzed by toluidine blue staining after 7 d. The apoptotic osteoclasts were analyzed by Annexin V-FITC staining after 2, 4, 8, 12, 24, 36, and 48 hours, to observe their apoptosis. Their total RNAs were extracted, and analyzed for TRAP and Cathepsin K expressions by Real-time RT-PCR., Result: Compared with the control group, both of the PNG group and the NG group showed much less osteoclasts (TRAP positive cells), lower TRAP activity and TRAP and Cathepin K (CTSK) expression, and smaller number of bone resorption pits and areas. The PNG group show lower indexes than the NG group. Additionally, the PNG group reached the apoptotic peak of osteoclasts at 12 h after drug administration, whereas the NG group reached after 24 h. And the former had more apoptotic cells than the latter., Conclusion: 8-PNG is much more active than NG in inhibiting the resorption of osteoclasts and inducing apoptosis of osteoclasts. Their only difference lies in 8-prenyl perssad, which is proved to be able to enhance the anti-bone resorption activity of 8-prenylnarigenin.

Objective: To observe antisepsis, anti-swelling, and therapeutic effects of Fuxiye (FXY), a Chinese medical lotion for external wash in treating vaginitis model rats., Methods: The cervicitis rat model was induced by agar plate diffusion, ear auricle swelling induced by dimethylbenzene, and chemical stimulus. The in vitro antibiotic actions of FXY were observed. Besides, its effects on the swelling and inflammation in model rats were also observed., Results: FXY at 25 mg/mL could completely inhibit the growth of Pseudomonas aeruginosa, Escherichia coli, pyogenic Streptococcus, and Streptococcus agalactiae. FXY at 50 mg/mL could completely inhibit the growth of Staphylococcus aureus and Candida albicans. It obviously restrained dimethylbenzene induced ear auricle swelling. It significantly alleviated cervicitis induced by chemical stiumli., Conclusion: FXY showed better effects on antisepsis, anti-inflammation, and treating cervicitis.

Objective: To investigate the outcome of the children with thoracic spine tuberculosis who underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach., Methods: From June 2005 to December 2010,9 children with thoracic spine tuberculosis underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach including 7 males and 2 females with an average age of 7 years old ranging from 3 to 12 years. The course of the disease ranged from 3 months to 1 year (averaged 6 months). All the patients had continuous thoracic and back pain,intercostal nerve pain and kyphosis deformity, accompanied with low fever,night sweat and pathologic leanness. The predilection sites examined by X-ray, CT or MRI were in T4 to T9 segment. The kyphosis angle ranged from 35 degrees to 72 degrees (averaged 48.2 degrees) before surgery. The ASIA classification was as follows: 2 cases at grade B, 5 at grade C, 2 at grade D. All the patients underwent a standard one-stage operation via posterior approach. Radical debridement was performed, then iliac crest bone autograft or allograft was placed and transpedicular screw system internal fixation was done to reconstruct the spinal column. The change of kyphosis angle and fusion of bone grafting were reexamined by X-ray regularly. The neurological function were evaluated according to ASIA classification., Results: There was no injury of blood vessel or spinal cord during the surgery. Nine children were followed-up for 16 to 38 months (averaged 24 months). The tuberculosis symptoms disappeared after surgery and there was no tuberculosis recurrence,incision infection, sinus formation and internal fixation failure in any of these chiildren. ESR reexamination recovered normally. Bony fusion was obtained in all patients and internal fixation position was normal 4 to 8 month postoperatively. The kyphosis angle ranged from 12 degrees to 30 degrees (averaged 19.50) at final followed-up. The function of spinal cord improved postoperatively, the function of spinal cord recovered at different degrees: 2 cases at grade C, 2 at grade D, and 5 at grade E., Conclusion: The one-stage posterior approach can provide direct and safe access to the lesion. The effect of vertebral canal decompression and kyphosis deformity correction were significantly. The structural iliac crest autograft or allograft and posterior transpedicular screw system could work effectively to stabilize the thoracic junction.

Objective: To observe the curative effects and complications of Bryan cervical disc replacement for cervical disc herniation., Methods: From Jannary 2005 to December 2008,39 patients with cervical disc herniation were treated with Bryan cervical disc replacement. There were 20 males and 19 females,with an average age of 47 years old (ranged, 35 to 59). Spinal compression symptom (20 cases) and nerve root symptom (19 cases) were main clinical symptoms. Single level disc was replaced in 35 cases and two-level replaced in 4 cases. Offset and activity of prosthesis,cervical physiological curvature, heterotopic ossification, prosthetic fusion were observed by dynamic X-ray. According to Odom's standard and JOA score,nerve function were evaluated; and depending on NDI standard,clinical symptom and daily function status were recorded., Results: All the patients were followed up from 16 to 36 months with an average of 24 months. Nerve function obviously improved and radiating pain of upper limb completely relieved. No patient with prosthetic anterior-posterior offset more than 2 mm was found. Prosthetic flexion and extention angle was (8.5+-1.8)degrees,left and right flexion range respectively were (3.5+/-1.2)degrees and (3.3+/-1.5)degrees. Cervical physiological curvature improved obviously or recovered normally. Three cases occurred in heterotopic ossification and 2 cases occurred in prosthetic fusion. According to Odom's standard,25 cases got an excellent results,9 good, 5 fair, the rate of excellent and good was 87.2%. JOA score increased from preoperative (8.26+/-1.32) to (15.71+/-1.89) at final follow-up and NDI decreased from preoperative (43.7+/-3.8) to (20.1+/-2.9) at final follow-up., Conclusion: Treatment of cervical disc herniation with Bryan cervical disc replacement can get the good curative effects,which can obtain good nerve functional recovery,cervical stability and activity. Nevertheless,the operation has typical complication such as heterotopic ossification and prosthetic fusion. Thus,it is important in chosing indication and operative procedure.

Percutaneous vertebroplasty(PVP), among various other options,has become a mainstay in the management of osteoporotic compression vertebral fractures. The purpose of this article is to review complications arising from the procedure and describes methods to minimize them. Complications can be classified as mild,which may include a temporary increase in pain; transient hypotension and cement leakage in the intervertebral disc space or into paravertebral soft tissues, moderate, including infection; extravasation of cement into the foraminal or epidural space and severe such as cement leakage in the paravertebral veins, leading to pulmonary embolism, cardiac perforation, cerebral embolism or even death.

Previous studies have found that 8-prenylflavonoids have a higher osteogenic activity than do flavonoids, which suggested that the 8-prenyl group may play an active role in bone-protective properties. To address this hypothesis, activities of 8-prenylnaringenin (PNG) and naringenin (NG) in osteoblast and osteoclast differentiation and function were compared in vitro. PNG was found to have a stronger ability than NG to improve osteoblast differentiation and osteogenic function in cultured rat calvarial osteoblasts, as demonstrated by levels of alkaline phosphatase activity, osteocalcin, calcium deposition, and the number and area of mineralized bone nodules, as well as mRNA expression of osteogenesis-related genes Bmp-2, OSX, and Runx-2. In addition, although expression of osteoclastogenic inducer receptor activator of nuclear factor kappa-B ligand (RANKL) was not affected, that of osteoclastogenesis inhibitor osteoprotegerin (OPG) and consequently the OPG/RANKL ratio were increased, more potently by PNG than NG. PNG was also found to have a higher potency than NG in inhibiting the osteoclast formation in rabbit bone marrow cells and their resorptive activity, as revealed by lower numbers of osteoclasts formed, lower numbers and areas of bone resorption pits, and lower mRNA expression levels of tartrate-resistant acid phosphatase and cathepsin K. Furthermore, PNG induced apoptosis of mature osteoclasts at a higher degree and at an earlier time than did NG. These results indicate that the 8-prenyl group plays an important role and contributes to the higher bone-protective activity of PNG in comparison with NG.

Objective: To 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., Methods: The 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., Results: Compared 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., Conclusion: Exposure 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.

A number of recent studies have suggested that flavonols (a class of phytochemical with many biological activities), might exert protective effects against post-menopausal bone loss. In the present study, we compared naringenin (NG) and 8-prenylnaringenin (PNG), two major naturally occurring flavonols, on in vitro differentiation of osteoblasts and bone resorbing activity, of rat bone marrow stromal cells (BMSCs). Our results indicated that both compounds, at 10(-6)  m, enhanced BMSCs' differentiation. Then effects of the two compounds at 10(-6)  m on ALP activity, osteocalcin secretion and calcium deposition, were compared over a time course. Numbers and areas of colonies stained for ALP (CFU-F(ALP) ) expression, and mineralized bone nodules, were histochemically analysed after 12 days and 16 days osteogenic induction, respectively. Expression of BMP-2, OPG, OSX, RUNX-2 genes and p38MAPK protein were examined using real-time PCR and western blotting, respectively. The data presented indicate that PNG, significantly enhanced the rat BMSCs' differentiation and mineralization through the BMP-2/p38MAPK/Runx2/Osterix signal pathway, greater than did NG. In conclusion, PNG has a more pronounced ability to enhance osteoblast differentiation and mineralization, than NG., (© 2012 Blackwell Publishing Ltd.)

Objective: To investigate the effect of exposure to static magnetic fields (SMFs) of 3.9 mT on proliferation and differentiation of osteoblasts in vitro., Methods: The 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., Results: Contrast 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., Conclusion: The cells exposed to the SMFs of 3.9 mT at 2.5 h apparently promote proliferation and differentiation of osteoblasts in vitro.

Objective: To investigated the effect of 50-Hz 3.6-mT sinusoidal electromagnetic fields (SEMFs) on the proliferation and differentiation of osteoblasts in vitro., Methods: The 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., Results: The 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., Conclusion: The SEMFs at 50-Hz 3.6-mT could suppress the proliferation of osteoblasts maturation but stimulate the differentiation and maturation of osteoblasts in vitro.

This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.

Objective: To investigate the effect of sinusoidal electricity magnetic fields (SEMFs) on the proliferation and differentiation of osteoblasts in vitro., Methods: Calvarial osteoblasts of newborn rats were isolated by enzyme digestion and randomly divided into 3 groups after subculture. Two groups of cells were exposed to 50 Hz 1.8 mT SEMFs for 30 min/d in parallel and vertical, respectively. Those without SEMFs exposure served as control. The cells were observed under the contrast phase microscope each day. After 48 h, cell proliferation was assayed by MTT method. The alkaline phasphatase (ALP) activities and calcium contents were measured after 3, 6, 9, and 12 days. The ALP positive colonies were histochemically stained after 10 days and the calcified nodules were stained by Alizarin Bordeaux after 12 days. Expressions of ALP, bone morphogenetic protein-2 (BMP-2) and Osterix (OSX) mRNA were measured at 0 h, 24 h, 48 h and 96 h., Results: The cells exposed to the SEMFs were arranged in spiral appearance after 3 days. Compared with control, SEMFs inhibited cell proliferation (P < 0.01 or P < 0.05), but enhanced the maturation and mineralization of the osteoblasts. The results showed that SEMFs improved ALP activities, promoted calcium contents, increased calcified nodulues numbers, boosted expressions of ALP, BMP-2 and OSX mRNA. SEMFs with magnetic lines of force in parallel has stronger activities than those in vertical., Conclusion: The SEMFs at 1.8 mT and 50 Hz inhibit the proliferation of osteoblasts, but enhance the maturation and mineralization of osteoblasts.

Objective: To investigate the outcome of radical debridement, reconstruction with bone autograft or allograft and plate internal fixation via the anterior approach for the treatment of cervicothoracic tuberculosis., Methods: From Jun. 2000 to Dec. 2010, 20 patients with cervicothoracic tuberculosis were treated by debridement and bone grafting with internal fixation via the anterior approach. They included 17 males and 3 females who ranged in age from 25 to 46 years (mean 38 years). The course of disease ranged from 3 months to 2 years (mean 12 months). The onset of the disease was chronic in all patients, with main complaints of persistent pain, and cervical stiffness and deformity accompanied with low fever,night sweating and pathologic leanness. Preoperative X-ray, CT or MRI showed that the pathologic change occurred in C7-T1 segment in 10 cases, T1 segment in 6 cases, T1-T3 segment in 3 cases,and T2-T3 segment in 1 case. The Cobb angle ranged from 25 degrees to 60 degrees (mean 35 degrees) before surgery. The Frankel classification was as follows: 2 cases at grade A, 4 cases at grade B, 7 cases at grade C, 2 cases at grade D, and 5 cases at grade E. All the patients underwent a standard cervical approach by combined partial median steotomy and transverse steotomy through the synostosis between the manubrium and body of the sternum to expose the lesion adequately. Radical debridement was performed, and then a tricortical iliac crest bone autograft or allograft was placed and secured by internal fixation to reconstruct the spinal column. The change in Cobb angle and fusion of bone grafting were reexamined by X-ray regularly. The clinical symptoms and neurological function were evaluated according to NDI (neck disability index) score and Frankel classification., Results: There was no injury to blood vessels, spinal cord or recurrent nerve during surgery. All patients were followed-up from 16 to 39 (mean 25) months. The tuberculosis symptoms disappeared after surgery and there was no tuberculosis recurrence,incision infection, sinus formation and internal fixation failure in any of these patients. ESR re-examination recovered normally. Bony fusion was obtained in all patients and internal fixation position was normal at 3 to 6 month postoperatively. The Cobb angle ranged from 10 degrees to 16 degrees (mean 12 degrees) and NDI was reduced from (48.2 +/- 2.9) to (22.5 +/- 3.1) at the final followed-up. Except for 2 patients at grade A showing no recovery preoperatively, the Frankel classification of the other patients raised 1.5 grade on average at the final followed-up, and the nerve function of the spinal cord recovered at different degrees: 2 at grade A, 1 at grade B, 1 at grade C, 3 at grade D, and 13 at grade E., Conclusion: The anterior approach can provide direct and safe access to the lesion. The decompression effect of the vertebral canal is significant. The structural iliac crest autograft or allograft and anterior instrumentation could work effectively to stabilize the cervicothoracic junction.

Nonunion of the tibia fracture may be caused by local infection, soft tissue defect,bone defect,fracture malaignment,bone marrow cavity block and degree of injury and so on. For less complications, central bone grafting is better than other methods for the treatment of selected nonunions of the tibia fracture. This procedure is performed through lateral approach, anterior to the fibula. Fresh autogenous bone from the iliac crest is used to form a central bridge between the tibia and fibula and the nonunion of the tibia at top and below. Application of internal fixation is beneficial to correct deformity and promote fracture healing. Central bone grafting is a safe and effective treatment for nonunions of the tibia.

Objective: To study the protective effect of genistein on osteoblasts treated with hypoxia., Methods: Rat osteoblasts were isolated from calvarias of newborn Sprague-Dawly rat by enzyme digestion and hypoxic environment was made by triple-gases incubator. Rat osteoblasts treated with hypoxia for 36 haurs. After 36 hours, cell viability, content of reactive oxygen species (ROS), analysis of cellular cycle and apoptosis, expression of proliferating cell nuclear antigen (PCNA), activity of iNOS and area of calcified nodules were detected. Total RNA was isolated and the gene expression of hypoxia inducible factor-1alpha (HIF-1alpha), BCL-2 and Caspase-3 was investigated by Real Time RT-PCR., Results: Genistein could significantly improve cell viability, percentage of G1 phases, area of calcified nodules and decrease apoptosis rate, ROS content, expression of PCNA, activity of iNOS. Besides, mRNA levels of HIF-1alpha and BCL-2 were enhanced and that of Caspase-3 was inhibited., Conclusion: Genistein can protect osteoblasts from hypoxia and enhance osteogenic differentiation significantly.

The objective of this study was to investigate the outcome of the patients with cervicothoracic tuberculosis who underwent radical debridement, reconstruction with bone autograft or allograft, and plate internal fixation via anterior approach. From June 2000 to December 2010, 20 patients with tuberculosis in cervicothoracic junction underwent a standard cervical approach, which was combined with a partial median osteotomy and transverse osteotomy through the synostosis between the manubrium and body of the sternum to expose the lesion adequately. Radical debridement was performed, then tricortical iliac crest bone autograft or allograft was placed and internal fixation was done to reconstruct the spinal column. The pathologic change regions were as follows: 10 in the C7-T1 segment, 6 in the T1 segment, 3 in the T1-T3 segment, and 1 in the T2-T3 segment. The classifications of Frankel were as follows: 2 at grade A, 4 at grade B, 7 at grade C, 2 at grade D, and 5 at grade E. There was no injury of blood vessel, spinal cord, or recurrent nerve during the surgery. The follow-up period ranged approximately 16 to 39 months. Bony fusion was obtained in all patients, and there was no internal fixation failure and tuberculosis recurrence in any of these patients. The nerve function of the spinal cord recovered at different degrees: 2 at grade A, 1 at grade B, 1 at grade C, 3 at grade D, and 13 at grade E. The anterior approach can provide direct and safe access to the lesion. The structural iliac crest autograft or allograft and anterior instrumentation could work effectively to stabilize the cervicothoracic junction.

Electromagnetic fields (EMFs) have been used clinically to slow down osteoporosis and promote fracture healing for many years. However, the underlying action mechanisms and optimal parameters of the EMF applications are unclear. In this study, we investigated the effects of treatment for different durations with 50 Hz sinusoidal electromagnetic fields (SEMFs) at different intensities on proliferation, differentiation and mineralization potentials of rat osteoblasts. Osteoblasts isolated from neonatal rats were treated with SEMFs (50 Hz at 0.9 mT-4.8 mT, 0.3 mT interval, 30 min/day up to 15 days). Compared to untreated control, SEMFs inhibited osteoblast proliferation (after 3 days' treatment) but increased alkaline phosphatase (ALP) activity (after treatment for 9 days) from 0.9 mT to 1.8 mT, declined from 1.8 mT until 3.0 mT, and then increased again from 3.0 mT to 3.6 mT and decreased once again from 3.6 mT to 4.8 mT. Numbers of colonies stained positive for ALP after 8 days and mineralized nodules stained by Alizarin red after 10 days showed the same bimodal tendency as with the ALP activity, with two peaks at 1.8 mT and 3.6 mT. SEMFs also bimodally increased Runx-2, Col1α2 and Bmp-2 mRNA expression levels in osteoblasts at 12, 24 and 96 h after exposure. The results indicated that while exposure to 50 Hz SEMFs inhibits the osteoblast proliferation, it significantly promotes differentiation and mineralization potentials of osteoblasts in an intensity-dependent manner with peak activity at 1.8 mT and 3.6 mT., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objective: To summarize the clinical application results of the micro-decompression procedure for the treatment of lumbar spinal stenosis with multilevel., Methods: From January 2004 to December 2008, 40 patients with lumbar spinal stenosis with multilevel were treated by micro-decompression procedure. There were 28 males and 12 females, ranging in ages from 55 to 80 years,with an average of 58 years. The course of this disease ranged from 18 months to 12 years,averaged 32 months. Forty patients with lumbar spinal stenosis with multilevel were diagnosed by CT or MRI examination. There were 20 cases with two levels stenosis (L4,5 and L5S1), 15 cases with three levels stenosis (L3,4, L4,5 and L5S1), and 5 cases with four levels stenosis (L2,3, L3,4, L4,5 and L5S1). The therapeutic effects were evaluated according MacNab standard in aspect of pain, bladder function, range of lumbar spine motion and muscle strength of lower limb., Results: All of the incisions healed without infections and complications. The mean operation time of each side was 70 minutes (ranged from 50 to 90 minutes), and mean blood loss was 150 ml (ranged from 90 to 200 ml). All the patients were followed up from 22 to 52 months with an average of 26 months. According to MacNab standard, 28 cases got an excellent result, 10 good and 2 poor., Conclusion: Operative treatment for lumbar spinal stenosis with multilevel should focus on the symptom sites causing by neural compression and preventive decompression operations are not necessary for nonsymptomatic sites. The micro-decompression procedure can be easily tolerated by older patients;it can decrease the damage to the posterior stabilizing structures of the lumbar spine. It is easily to access to spinal canal and decompress the nerve roots.

Fracture of the subtrochanteric area of the femur is one of the most difficult about the hip to manage. Many of these fractures are comminuted and result from high-energy trauma. Nonoperative treatment of these fractures may result in a significant rate of malunion, nonunion and other complications. Therefore, subtrochanteric fractures is preferred to be treated with operative methods. The Russell-Taylor classification is useful in planning the type of internal fixation. For fractures located below the level of the lesser trochanter, standard locked intramedullary nails can be used effectively. For fractures that extend into the lesser trochanter but do not involve the piriformis fossa, the options of a cephalomedullary nail versus a 95 degree fixed angle device have yielded the best results. For fractures that have proximal trochanteric extension into the area of the piriformis fossa, sliding nail screw devices may have some usefulness. The DHS implant is not employed for the treatment of subtrochanteric femoral fractures.

Objective: To investigate the effect on early treatment with vacuum-assisted closure(VAC) to wound healing of acute explosion injury in pigs, and provide a new way for early treatment of battle wounds., Methods: Eight healthy 3-month Landrace pigs of both sexes with the body mass of (50 +/- 5) kg were selected in the study. Sixteen battle wounds were made by explosion of same type detonator (pattern number: 660929F48840-55, included DDNP 0.3 g, RDX 0.7 g) in hibateral skin of buttock of 8 pigs, which were divided into experimental group and control group (pair wounds of left and right). The raw sufaces were thorough debrided at 3 h after exposure, according to the characteristics of treatment on the battlefield, experimental group was treated with VAC under the pressure of (-50 +/- 5) Kpa after debridement and sterilization and control group was treated with routine dry sterile gauze draping. Results of bacteriology (bacterial counts and the proportion of G+ bacteria) and pathology (HE stain and Masson stain) were detected at every wound before and after treatment., Results: At the 3 days after treatment,the bacterial number in the experimental group was [(7.82 +/- 0.55) x 10(4) ] CFU/g, in control group was [(1.07 +/- 0.14) x 10(6)] CFU/g. There was significant difference between two groups. The proportion of G+ bacteria in experimental group was significantly increased. The raw surface in experimental group was clean with affluent and neoformative granulation tissue, blood vessels and collagen, necrotic tissue decreased obviously by pathological observation., Conclusion: VAC could reduce the quantity of bacteria, improve the proportion of G+ bacteria, and promote the formation of granulation tissue and the healing of wound. The VAC for the treatment of battle wounds has a positive effect.

Objective: To prepare macroporous bone substitute composed of calcium phosphate cements and rhBMP-2 loaded gelatin microspheres, and to investigate ectopic osteoinduction of the composite., Methods: After being prepared by improved emulsified cold-condensation method and crosslinked by 5% genipin solution,gelatin microspheres (GMs) were observed by scanning electron microscope (SEM) and loaded with rhBMP-2 by adsorption. Macroporous bone substitute was developed by mixing calcium phosphate cement (CPC) with 2.5% GMs, being as the experimental group,and CPC with rhBMP-2 was the control group. After the both composites had been soaked in the sodium chloride for 1 week or 3 weeks, compressive strength of the composites were tested, and the cross-sections were observed by SEM. Concentrations of rhBMP-2 in the solutions at different time by ELISA method and the cumulative drug release amount was calculated. The composites had been implanted in the muscle bags of the mouses for 3 weeks. Then the tissues around the materials were collected, stained by hematoxylin and eosin, and Ca and ALP in the tissues were also measured., Results: Gelatin microspheres were spherical with diameters of (62 +/- 18) microm. Macropores appeared in the experimental materials 1 week and 3 weeks after being soaked,and total porosity, macroporosity, cumulative release amount of rhBMP-2 in the experimental group were higher than that in the control. But compressive strength of the experimental group was lower than that of the control group 3 weeks after being soaked. Results of HE stain showed chondral formation in both groups, but there were more chondral tissues in the experiment group, and so were the concentrations of Ca and ALP., Conclusion: Macroporous calcium phosphate cement can be prepared by using rhBMP-2 loaded gelatin microspheres, and it is an excellent bone substitute due to it's proterty of promoting rhBMP release and powerful ectopic osteoinduction.

This study is to investigate the effects on the expression of iNOS and production of NO in the osteogenic differentiation process of rat bone marrow stromal cells (rBMSCs) by icariside II. rBMSCs were cultured by adherence screening method. When the culture dishes were covered with 80% cells, the osteogenic induced cultures were adopted. Icariside II was supplemented into the culture at 1 x 10(-5) mol x L(-1). The activity of iNOS, content of NO and osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-FALP and mineralized bone nodules were compared among the icariside II-supplemented group, L-NMAE group, icariside II + L-NAME group and the control. Total RNA was isolated and the gene expression of iNOS, Osterix and Runx-2 was investigated by real-time PCR. Total protein was also isolated and the secretion of iNOS and collagen I was examined by Western blotting. Icariside II can significantly improved ALP activity, CFU-FALP amount and mineralized nodules. Besides, the mRNA level of factors related to the osteogenic differentiation includes Osterix and Runx-2 also enhanced. The secretion of collagen I also promoted significantly. But all of these effects can be inhibited by L-NAME which can specifically inhibit the activity of iNOS. Icariside II enhances the osteogenic differentiation of rBMSCs significantly, but if the activity of iNOS was blocked by L-NAME, the osteogenic differentiation markers decrease accompanied with iNOS and NO decrease, suggesting that icariside II stimulates the osteogenic differentiation via enhancing the activity of iNOS and promoting the generation of NO.

There has been a strong interest in searching for natural therapies for osteoporosis. Genistein, an isoflavone abundant in soy, and icariin, a prenylated flavonol glycoside isolated from Epimedium Herb, have both been identified to exert beneficial effects in preventing postmenopausal bone loss. However, the relative potency in osteogenesis between the individual phytoestrogen flavonoids remains unknown. The present study compared ability of genistein and icariin in enhancing differentiation and mineralization of cultured rat calvarial osteoblasts in vitro. Dose-dependent studies in osteoblast differentiation measuring alkaline phosphatase (ALP) activity revealed optimal concentrations of genistein and icarrin for stimulating osteogenesis to be both at 10(-5) M. Time course studies comparing the two compounds both at 10(-5) M demonstrated that icariin treatment always produced higher ALP activity, more and larger areas of CFU-F(ALP) colonies and mineralized nodules, more osteocalcin secretion, and calcium deposition, and a higher level of mRNA expression of osteogenesis-related genes COL1α2, BMP-2, OSX, and RUNX-2. However, they inhibited the proliferation of osteoblasts to a similar degree. In conclusion, although future in vivo studies are required to investigate whether icariin is more efficient in improving bone mass and/or preventing bone loss, our in vitro studies have demonstrated that icariin has a stronger osteogenic activity than genistein. In addition, while the prenyl group on C-8 of icariin could be the active group that takes part in osteoblastic differentiation and explains its greater potency in osteogenesis, mechanisms of action, and reasons for the relative potency of icariin versus genistein need to be further studied., (Copyright © 2010 Wiley-Liss, Inc.)

Objective: To investigate the effect of isopsoralen on proliferation, osteogenic differentiation and calcification capacity of rat calvarial osteoblasts (ROB)., Methods: Segregated neonatal SD rat skull,and digestion with enzyme to obtain bone cells and cultured in MEM containing 10% FBS. Exchange the medium after three days, proceeded serial subcultivation when cells covered with 90% culture dish. Proliferation analysis was performed in 96-well plates use MTT method, isopsoralen's final concentration were 1 x 10(-4), 1 x10(-5), 1 x 10(-6), 1 x 10(-7) mmol/L. Differentiation analysis was performed in 24-well plates, the Alkaline phosphatase activity and calcium salt sediment yield and osteocalcin measured at the 4th, 8th, 12th, 16th day. At 12th day, proceeded ALP stain, and at 14th day for alizarin red staining and calcified nodule count., Results: When the Isopsoralen's final concentration was 1 x 10(-5) mmol/L, there was no significant effect on the ROB's proliferation, but it could promote osteogenesis. It also could raise the ALP activity and calcium salt sediment yield and osteocalcin, increase calcified tubercle amount., Conclusion: When the isopsoralen final concentration is 1 x 10(-5) mmol/L, it promoted ROB differentiation and maturation. Isopsoralen may be the active ingredients of preventing anti-osteoporosis in Psoralea corylifolia.

Objective: To investigate the effects of icariin and it's main metabolites-icariside II on the osteogenic differentiation of rat bone marrow stromal cells (rBMSCs)., Methods: rBMSCs were cultured by adherence screening method, icariin and icariside II were supplemented into the culture at 5 x 10(-5) mol/L respectively. The osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-F(ALp), osteocalcin secretion, calcium deposition and mineralized bone modulus were compared among the icariin-supplemented group, icariside II and the control. The gene expressions of bFGF, IGF-1, Osterix and Runx-2 were examined by RT-Real Time PCR., Results: Both icariside II and icariin significantly improved ALP activity, CFU-F(ALP) amount, osteocalcin secretion, calcium deposition and mineralized modulus. Besides, they enhanced the gene expressions of bFGF, IGF-1, Osterix and Runx-2. Icariside II was obviously stronger than icariin at the above activities., Conclusion: Icariside II is stronger than icariin at enhancing the osteogenic differentiation of rBMSCs, suggesting that icariin can be administered via oral and it's metabolites are the effective constitutes for antiosteoporosis activity.

Objective: To investigate the effects of Osthol on the proliferation and differentiation of osteoblasts of rats (rat calvarial osteoblasts, ROB) cultured in vitro., Methods: The neonatal SD rat skull was segregated, and enzyme digestion was used to obtain bone cells which were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subcultivation was performed when cells covered with 90% of the culture dish. The Osthol was added to 96-well plates with final concentration of 1 x 10(-4) mol/L, 1 x 10(-5) mol/L, 1 x l0(-6) mol/L and 1 x10(-7) mol/L, and MTT method was used to evaluate the proliferation. Differentiation analysis: the alkaline phosphatase (ALP) activity was determined at the 3rd, 6th, 9th, 12th and 15th days separately after osteogenic induction culture. The synthesis of type I collagen was observed using immunohistochemical method at the 8th day. The ALP stain was performed at the 12th day. The alizarin red staining was done and calcified nodules was counted at the 14th day., Results: The Osthol with final concentration of 1 x 10(-4) mo/L inhibit the proliferation of ROB. The Osthol with final concentration of 1 x 10(-5) mol/L had no obvious influence on the proliferation of ROB, but it significantly promoted the activity of ALP, enhanced the synthesis of collagen type I and increased the number of calcified nodules., Conclusion: The Osthol with final concentration of 1 x 10(-5) mol/L can promote differentiation and maturation of ROB, which may be active ingredients of Chinese drugs for the osteoporosis prophylaxis.

The Chinese investigators separated bullet wounds into three zones in the beginning of 1980s: a primary wound tract, a contusion zone adjacent to prinary wound tract, and a concussion zone neighboring the contusion zone. Basing on the research results by MRI scan and pathological observation, the author and his coworkers recently proposed that the gunshot wounds should be divided into four consecutive zones: a primary wound tract, a zone of coagulative necrosis, a zone of muscle disruption, and a zone of muscle distortion. A zone of coagulative necrosis plus a zone of muscle disruption equals to a contusion zone, they are separately named because the former is irreversibly devitalized and the latter still has the ability to recover. The sectionalized method shows the range of debridement and provide reference for the conservative treatment or thoroughly debridement. However, the mechanism of each zone need to be further studied.

The spine is the most common site of metastatic tumors. The highest incidence of metastatic tumors on the spine is in the lumbar region, followed by the thoracic and cervical levels. Most associated spinal cord lesions, however, are located in the thoracic spine. The goals of surgery is decompression of the neural tissues, prevention of neural injury by stabilization of the unstable spinal column,or pain control. When there is a need to decompress the neurologic tissues,the most direct approach is anterior because the most common site of metastatic tumors is in the vertebral body. Laminectomy alone is not effective in improving neurologic function. Nevertheless,the posterior approach is more accessible, less hazardous, and effective in the correction of deformity if combined with instrumentation. If complete vertebrectomy is feasible in anterior bone strut combined with anterior and posterior stabilization is necessary following the vertebrectomy.

Objective: To investigate the effect of pulsed electromagnetic fields (PEMs) on inducing osteoclastic like cell (OLC) formation changes and apoptosis in ovariectomized (OVX) rats bone marrow culture in vitro., Methods: Thirty healthy three-month-old female Wistar rats were either sham-operated (Sham) or ovariectomized (OVX) and randomly divided into three groups: group A (OVX + PEMs, 18 rats), group B (OVX, 6 rats) and group C (Sham, 6 rats); group A was again randomly divided into three groups: A1, A2, A3. The frequencies adopted were 1.5, 2, 75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months, then the bone marrow of all rats were cultured, 2 days later, group A cells (including group A1, A2, A3) were collected and exposed to different frequencies PEMs for 2 weeks (30 min/day). In order to observe the changes of osteoclasts and count their numbers, cells were taken for Wright Giemsa staining, tartrate-resistance acid phosphatase (TRAP) staining and Hoechst 33258 staining., Results: TRAP staining results indicated the number of OLC in group C was the least, then was group A2, A3, A1, B. The number of OLC in group B was remarkably increased (P < 0.01; vs group C, A2). The number of OLC in group B was significantly increased (P < 0.05; vs group A1, A3). Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups, which of group C, A2 was significantly increased (P < 0.05; vs group B)., Conclusion: PEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized (OVX) rats bone marrow culture in vitro, the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy.

Objective: To evaluate clinical result of the micro-decompression procedure for lumbar spinal stenosis., Methods: From September 2001 to May 2006,87 patients (male 60, female 27) with lumbar spinal stenosis underwent micro-decompression. The age of patients were from 43 to 80 years with an average of 51 years. Among them,2 cases with spinal stenosis occured in L(3,4), 47 in L(4,5), 38 in L5S1., Results: All patients were followed up for 18-48 months with an average of 26 months. The results were excellent in 52 cases, good in 28, poor in 7, according to Macnab of back leg pain standard. The rate of excellent and good was 92%., Conclusion: Operative treatment for lumbar spinal stenosis is focused at the areas causing symptomatic neural root compression rather than prophylactic decompression at areas of nonsymptomatic disease. The micro-decompression procedures are more likely to be well tolerated by older patients.

Locking plate technology offers improved fixation stability in osteopenic bone and for comminuted and periarticular fractures. The additional stability per screw compared with that of conventional nonlocking fixation enhances the application of minimally invasive fracture techniques. The application of locking plates is somewhat more difficult than the placement of conventional plates. Fracture reductions are often done indirectly, the locking screw must be carefully aligned along the axis of the receiving hole to ensure proper tightness, and the length of the plate must be selected carefully. The use of locking plates will likely increase, particularly with the increasing prevalence of osteopenic fractures on our aging population and the increase in high-energy fractures in younger patients severe trauma.