BACKGROUND: There were disadvantages of animal models in the study of the relationship of muscle and skeleton. Recent study has been demonstrated the effect of Botulinum toxin type A to atrophy local muscle without influencing the surrounding muscle.OBJECTIVE: To construct a reasonable animal model of local muscle atrophy by local injection of Botulinum toxin type A.METHODS: Totally 25 male Wistar rats of 4 months were subjected to ketamine (0.2 mL/kg) and Sumianxin (0.2 mL/kg) by intramuscular injection. A 0.5-cm incision was made on the middle of dorsal femur to expose quadriceps femoris. Right quaddceps femods was injected with 2 Units (0.2 mL) of Botulinum toxin type A, and left quaddcaps femoris of the same rat with the same amount of saline as controls. At 1, 2, 4, 8, 8 weeks after injection, 5 rats were used to take gross observation and histological examination.RESULTS AND CONCLUSION: Gross observation of the muscle tissue showed that, compared with self-controlled group, the volume and weight of the quadriceps femods were decreased significantly (P < 0.01). The histological examination of muscle tissue showed the atrophy of the quadricaps femods from expedmental group was more obvious, the muscle fiber become thin,and the nuclei of the muscle fiber assemble together, with small distance betWeen muscle fibers. Weight of the quadricaps femods treated with Botulinum toxin type A was decreased at 1 and 2 weeks. The increase in weight of muscle was slow among muscle at 4, 6 and 8 weeks. The muscle weight showed an increased tendency in the saline side at vadous time points. Injecting Botulinum toxin type A into local muscle is a reasonable way to set up an experimental model of the atrophy of a destination muscle with strong practice, good repeatability, high stability, and may be used to examine the relationship of muscle and skeleton.

Objective To observe the ultrastructural alteration and healing characteristics of osteoporotic fracture, and to elucidate its cellular mode of healing. Methods Eighty female SD rats of 8 months old, which had the weight of 290 to 340 g, were randomized into two groups of 40 each: the osteoporotic fracture model(OPFM) and the common fracture model(CFM). After anesthesia, the bilateral posterior transperitoneal approach was performed in the OPFM group and the bilateral ovaries were removed; and in the CFM group, only the sham operations were performed. Three months later, the fracture of femoral middle shaft were created and fixed with a Kirschner pin through medullary cavity. The callus of each rat was examined by transmission electron microscopy(TEM) and scanning electron microscopy(SEM) in 5 days and 1, 2, 4, 6, 8, 12 and 16 weeks postoperatively. Results TEM showed that the number of chondrocyte in OPFM group was greater, but function was lower than that of CFM group. Volume of collagen secreted by the chondrocyte was less and arranged irregularly during the fracture healing period in OPFM group. Number and function of osteoblasts in OPFM group were lower than that of CFM group. Extracellular collagen was disordered and sparse, but function of osteoclasts in OPFM group was more active than that of CFM group. The SEM showed that the collagen in callus of CFM group was dense and arranged in good order or pyknotic. Conclusion The fracture healing of the osteoporotic fractures is due to the decrease of the osteoblastic formation and the increase of the osteoclastic resorption as well as the poor bony healing quality.

Objective To observe the expression of platelet-derived growth factor A(PDGF-A) and PDGF-?R in callus during osteoporotic fracture healing and to explore further into the mechanism or effect of PDGF-A and PDGF-?R on osteoporotic fracture healing. Methods The expression and change of PDGF-A and PDGF-?R in different period of osteoporotic fracture healing(5, 7, 14, 28, 42 days) were investigated by means of immunohistochemistry(ABC method). Results There were different cells origin (chondrocyte, osteoblast, osteocyte, vascular endothelial cell, et al) and degree of expression of PDGF-A and PDGF-?R in callus during different period of osteoporotic fracture healing. Conclusion PDGF-A moderates and participates in osteoporotic fracture healing. The decrease of osteoporotic fracture repair capacity may correlate with abnormality of PDGF-A secretion.

OBJECTIVE: To study the effect of melittin on apoptsis and necrosis of osteosarcoma cell line U2 OS in vitro. METHODS: Osteosarcoma cell line U2 OS was treated with melittin. The growth and proliferation was observed by MTT assay and cell counting, and the necrosis was estimated by Trypan blue staining. The cell apoptsis, Fas and Apo2. 7 expression were detected by cytometer. RESULTS: The data showed that melittin could inhibit the proliferation of U2 OS dose-dependently at 16 and 64 mg/L. Cell apoptsis was detected by cytometer, when the cells were treated by 16 mg/L and 32 mg/L of melittin respectively, and the percentages of Fas and Apo2. 7 positive cells were increased. CONCLUSION: Melittin inhibits the proliferation of osterosarcoma cell line through up-regulating Fas expression and inducing apoptsis.

[Objective]To observe the effect and the safety of the COX-2 inhibitor Celecoxib as the analgesia in orthopaedic post-operative patients.[Method]Sixty-four operative patients were selected in all between 2004～2005 in hospital,and divided into two groups randomly.Celecoxib was used as the postoperative analgesia in the experiment group and the "Patient-controlled analgesia" in the control group.The administration of Celecoxib: 8-12 hours before operation generally,just before the abrosia,and 6 hours after operation when patients can take food.drug withdrawal was depended on the complex of the operation and the severety of pain in 3 to 5 days.Dosage:400 mg first time,if a complicated operation,larger dose would be given.VAS score,adverse reaction and the satisfaction of patients were observed.[Result]The effect of Celecoxib is similar as the PCA,with less adverse reaction and more satisfaction.[Conclusion]Celecoxib has a satisfactory effect and safety as a postoperative analgesia,and is suitable to be a peri-operative analgesia of orthopaedic patients.

Objective To construct the recombinant plasmid of PA-ⅠL and express in E.coli BL 21 (DE3), and evaluate the biological activity of recombinant protein.Methods PA-ⅠL gene was amplified by PCR using primers designed according to Pseudomonas aeruginosa genome sequences and then cloned to the vector pET -28a ( +).The recombinant plasmid was transformed into E.coli BL21(DE3) and induced to express by IPTG.The recombinant protein was purified by nickel affinity chromatography.The binding activity of recombinant PA-ⅠL with Gb3/CD77 was evaluated by flow cytometry.The function of recombinant PA-ⅠL on the binding of bacteria with host cells was evaluated by colony plate counting.Results and Conclusion The recombinant PA-ⅠL protein was highly expressed in E.coli BL21(DE3) and protein purity by SDS-PAGE analysis was high after nickel affinity chromatography .Besides, the recombinant PA-ⅠL had binding activity to Gb3/CD77 and inhibited the binding of PAO1 to host cells in a dose-dependent manner.

BACKGROUND:Calcium sulfate has good biocompatibility and biodegradability, which is a safe and effective bone graft substitute.
OBJECTIVE:To investigate the osteogenesis ability of calcium sulfate combined with bone marrow mesenchymal stem cells.
METHODS:After L4/5 posterior lumbar discectomy, 36 rabbits were randomized into three groups:rabbits in autologous bone group were implanted with autologous iliac bone via the intervertebral space;animals in al ogenic bone group were implanted with decalcified bovine bone;rabbits in tissue-engineered bone group were implanted with calcium sulfate combined with bone marrow mesenchymal stem cells. Bone formation and molding were observed by gross observation, anteroposterior and lateral X-ray, histology and biomechanics at 4, 8 and 16 weeks. Cal us specimens were employed for histological observation of interbody fusion. Biomechanical analysis of spinal fusion site was conducted at 16 weeks.
RESULTS AND CONCLUSION:Sixteen weeks later, interbody fusion was complete in the autologous bone group, the trabecular bone bridged continuously and a large amount of woven bone was merged into pieces;in the al ogenic bone group, incomplete bony fusion was found between the intervertebral space, most of cartilage tissues differentiated into bone, but fibrous tissue was also ful of the central part;in the tissue-engineered bone group, interbody fusion was complete, and a large amount of woven bone was fused into pieces, while the artificial bone was absorbed and ossified with smal residual. Failure strength and stiffness in the autologous bone and tissue-engineered bone groups were superior to those in the al ogenic bone group. These findings indicate that the calcium sulfate/bone marrow mesenchymal stem cells tissue-engineered bone has excellent osteogenic and osteoinductive capacity that can exert a good function of promoting spinal interbody fusion.

Objective The brain volume and behavioral performance of emotional memory were measured by combining voxel-hased morphometry (VBM) and behavior to explore the morphological characters of brain regions related to the emotional enhancement effect of memory.Methods A total of 32 healthy young subjects participated in the study.The experimental processes included MR scan and behavioral performance of emotional memory the latter of which involved two phases-encoding and retrieval.Behavioral performance was recorded and the whole-brain 3D MRI data were acquired via 3.0T MR.3D MRI data were segmented by VBM5,and the wholebrain volumes of gray matter,white matter and CSF were produced.SPM5 and SPSS13.0 were used for statistical analysis of the 3D MRI data and behavioral data,respectively.Results Retrieval accuracy of the emotional pictures was higher than that of the neutral pictures ( t =5.08,P＜0.001 ),the emotional enhancement effect was significant ( Δ Pr =0.12 ± 0.01 ).Brain regions related to emotional enhancement effect included bilateral amygdala,dorsolateral prefrontal cortex,middle frontal gyrus,dorsomedial prefrontal cortex,fusiform gyms,right parahippocampus,left orbitofrontal cortex and middle cingulate gyms.Volume ratio of bilateral amygdala were related to emotion enhancement effect evidently (left:r=0.564,P＜0.01 ; right:r=0.541,P＜0.01 ).Conclusion This study morphologically confirms that brain regions,such as amygdale and prefrontal cortex,are critical structures for the enhancement of emotional memory,and further extends the findings of previous functional imaging studies.

Objective To analysis of risk factors for left atrial thrombosis in patients with rheumatic mitral stenosis.Methods From January 2001 to December 2008, 2277 patients with rheumatic mitral stenosis underwent operations in our hospital. There were 737 males and 1540 female, the age ranged from 19 to 84 years [average (50.9 ±10.2) years]. Left atrial thrombosis group (554 cases) and no thrombosis group (1723 cases) were divided, retrospectively collected data were analyzed with univariate and multivariate Logistic regression. Results 12 bvariables, including age, mitral valve orifice area, left atrial diameter, left ventricular diastole diameter, CRP, gender , degree of mitral stenosis, or regurgitation, degree of bicuspid regurgitation, degree of pulmonary hypertension, atrial fibrillation and heart function had statistic difference between two groups. With multivariate Logistic regression for these 12 factors, age, mitral valve orifice area, left atrial diameter, degree of mitral regurgitation and atrial fibrillation were found to be the affecting factors for left atrial thrombosis in patients with rheumatic mitral stenosis. Conclusion For patients with rheumatic mitral stenosis, age, mitral valve orifice area, left atrial diameter and atrial fibrillation are the risk factors for left atrial thrombosis. Mitral regurgitation is a protective factor for left atrial thrombosis.

Fifty-four children, aged 1.5-3.5 years, were selected from a nursery and divided into three groups. To the children in group 1 multivitamin fortified formular milk powder of Hai-He brand (70g daily) was given as a supplementary food in addition to the normal diet and whole milk powder of Hai-He brand was given to group 2 as another supplementary food. The third group served as control. This observation was lasted for three months. At the end of the third month, the increments of body weight of the children, tricep skinfold and circumference of upper arm of group 1 were higher than those of group 3 significantly, but there were no significant differences between group 1 and group 2. The hemoglobin content of group 1 increased by 0.8g/dl but group 2 and 3 decreased by 0.53g/dl and 0.72g/dl respectively. At the end of our observation, the concentrations of vitamin B1, C and PP in urine of group 1 were higher than those of other two groups by vitamin load test.Thus, we may consider that the multivitamin fortified milk powder is better than the whole milk powder in improving the vitamin and iron nutritional status of young children.