[Objective]To study the effects of verapamil on the immediate-early gene(IEGs)expression of bone marrow mesenchymal stimulated by mechanical strain,and deduce the effects of calcium ion channel on the early responses of children bone marrow mesenchymal stem cells(MSCs)to mechanical strain in vitro.[Method]MSCs were isolated and cultured.The 3-6th generational MSCs were stimulated in different time by mechanical and(or)verapamil(20?mol/l).Flow cytometry was applied to measure intracellular free Ca2+ at once.Early-response genes /proteins(c-fos,c-jun and c-myc)were examined by RT-PCR and immunocytochemical stain.[Result]The application of mechanical strain increased intracellular c-fos,c-jun and c-myc mRNA /protein levels.The resulting effect is partially inhibited before MSCs were preincubated with verapamil(P

Objective To explore the hospitalization expenses and its related factors of injury cases in Shenzhen city in 2006,and to provide basis for the expense control in future.Methods The medical expense proportion was calculated on 7 180 injury cases with hospitalization data from Shenzhen injury surveillance system,ANOVA and multivariate regression analysis were used to determine the influencing factors for the expenses.Result The medical expenses of injury inpatients in Shenzhen were 3 835.65 RMB/case and 437.86 RMB/case/day.36.10% and 20.27% of the total expenses were attributed to medication and therapy,respectively.The expense per day was correlated strongly with the level of hospital,length of stay,age,method of payment and injury location.Conclusions The disease burden of injury was higher than that of other diseases,Hospitalization expenses can be effectively controlled by taking strategies,such as enhancing the care and treatment before hospitalization,shortening the duration of hospitalization,avoiding infection in hospital and so on.

Objective To summarize the experience of ultrasound-guided percutaneous nephrostomy during percutaneous nephrolithotomy(PCNL) under B-ultrasound guidance. Methods A ultrasound-guided percutaneous nephrostomy was performed in 43 cases by using a 18-gauge needle.Then the percutaneous puncture was expanded for establishing a working tunnel. Under nephroscope,stone fragmentation was completed in the renal pelvis and the upper segment of the ureter.Results The success rate of percutaneous puncture was 100%.Of the 43 cases,the percutaneous working tunnel was successfully established in 42 cases(97.7%),while a conversion to open surgery was needed in 1 case because of uncontrolled hemorrhage after puncture.The overall stone clearance rate was 100%(42/42),including 38 cases of stone removal on one session and 4 cases of two-staged clearance.No serious complications,such as massive hemorrhage or neighboring organ injuries,were encountered.Re-examinations with B-ultrasonography or KUB at 1 month after operation showed no residual stones.Conclusions Ultrasound-guided percutaneous nephrostomy is minimally invasive,safe,and reliable.Proper use of this technique is crucial for a successful performance of percutaneous nephrolithotomy in the treatment of upper urinary tract calculus.

Objective To investigate the effects of protein active composition of Scorpio on apoptosis of L1210 tumor cells for the purpose of establishing the quality evaluation method of biological effect of Scorpio. Methods L1210 cells were examined by trypan blue exclusion. The proliferation of cells was determined by improved MTT assay. Flow cytometry was performed to analyze cell apoptosis with propidium iodide (PI). Results When the concentration of protein active composition of Scorpio exceeded or equaled 37 mg/ml, the coefficient correlation of the growth inhibiting curve of L1210 cells was 0.9357, and IC50 was 175 mg/ml. The excellence time was 0 to 48 hours. When the concentration of protein active composition of Scorpio exceeded or equaled 9.25 mg/ml, the apoptosis ratio of L1210 cells was raised significantly.Conclusion The protein active composition of Scorpio might promote the apoptosis and restrain the proliferation of L1210 cells. The value of anti-tumor biological effect of the protein active composition of Scorpio was 9.25 ～ 175 mg/ml. This value may be one of the indexes for quality evaluation of biological effect of Scorpio.

Objective:[WT5BZ]The effect of different contents of zinc in feed on learning and memory function and brain somatostatin,zinc and calcium concentrations in rats were studied. [WT5HZ]Methods:[WT5BZ]The feeds contained 20, 50, 100, 200, 400, 800 mg/kg zinc respectively.The learning and memory function of animals were determined by radioimmunoassay.Their zinc and calcium contents were determined by atomic absorption spectrophotometry. [WT5HZ]Results:[WT5BZ]Feeds containing 100 and 200 mg/kg zinc were adequate for learning and memory function of rats,whereas other feeds resulted in poor learning and memory function,and depression in somatostatin concentration in hypothalamus,hippocampus and cerebrum cortex.Additionally,feeds containing lower than 100 mg/kg zinc decreased zinc concentrations in blood,hippocampus,cerebrum cortex and hypothalamus,and calcium concentration in cerebrum cortex. [WT5HZ]Conclusion:[WT5BZ]These results suggest that 100 200 mg/kg zinc in feed is adequate for maintaining learning and memory function in rats.

Objective: Three experiments were used to evaluate the effects of zinc deficiency on growth, learning and memory ability of rats. [WT5FZ]Methods: In experiment Ⅰ and experiment Ⅱ, the experimental rats were divided into three groups, which were zinc deficiency group (ZD), pair fed group (PF), and zinc deficiency and supplementation group (ZS). Their durations were 35 days and 28 days respectively. In experiment Ⅲ, the experimental rats were divided into ZD, PF and ad libitum (AL) group. Results: Zinc deficiency significantly decreased the growth and learning ability of rats, whereas zinc supplementation reversed the above status. Conclusion: The results suggest that zinc not only influence growth, but also the advanced function like learning and memory ability of brain.

AIM: To investigate the details of CD4 + T cell polarized to Th1/Th2 in vitro . METHODS: After isolated the PBMCs and blood-plasma from adult human peripheral blood by Ficoll-Hypaque centrifugation, the PBMC culture procedure with or without the self-blood -plasma was applied to polarize T cells in vitro , these cells were polarized by PHA(20 mg/L),non-PHA respectively. The polarized rates of Th cell after 24 h,48 h,72 h were estimated respectively by flow cytometry following two-color immunofluorescent staining. RESULTS: CD4 +T cell would polarize to Th1/Th2 two subsets after self-cytokines and PHA activation in vitro . The polarized rates of T cell after cultured for 24 h,48 h and 72 h were (13.28%?1.59%)/(12.70%?1.65%),(17.19%?1.03%)/(17.50%?1.30%),(19.49%?2.87%)/(18.58%?1.49%) respectively, but the polarized rates of T cell were very low if without self-blood-plasma. The difference between them was significant. The ratios of Th1/Th2 cells were about 1. CONCLUSION: CD4 +T cell from adult human peripheral blood would polarize to Th1/Th2 two subsets in the presence of self-blood-plasma and PHA(20 mg/L) in vitro , and the cell number of Th1 and Th2 would be in balance.

Objective:To identify the risk factors for premature proximal femur physeal closure (PPC) in children treated surgically for femoral neck fractures.Methods:Data of 106 children with an open triradiate cartilage who were treated surgically for a femoral neck fracture were retrospectively analyzed. Age, gender, laterality, mechanism of injury, the type of fracture, initial displacement, time to reduction, fixation method, whether the implant crossed the physeal plate, reduction method, reduction quality and development of femoral head avascular necrosis (AVN) were collected. PPC of the proximal femur was assessed through postoperative 6-12 months radiographs.Results:A total of 106 patients with an open triradiate cartilage were followed up, with an average duration of 20.4±13.3 months (range, 6-86 months). The overall rate of PPC following paediatric femoral neck fractures treated surgically was 36.8% (39/106). Among the 39 patients with PPC, 25 were males and 14 were females; the average age at the time of injury was 9.7±3.6 years (range, 3-15 years); 23 patients were involved in left hips and 16 were in right; the mechanisms of injury included motor vehicle accident in 5 fractures, falling injury in 21, sports-related injury in 12 and other causes in 1 fracture; two hips were Delbet type I, and 26 hips and 11 hips were Delbet type II and III, separately; type II and III of initial displacement were involved in 26 and 13 patients, separately; the mean duration from injury to surgery was 3.3±2.8 d (range, 1-14 d); 2 hips were treated with Kirschner wires, 35 hips with screw fixation, and the remaining 2 hips underwent screw and plate fixation; 2 hips had the hardware crossing the proximal femoral growth plate, and the remaining 37 hips didn’t; 15 hips were treated by closed reduction and internal fixation, and the other 24 hips underwent open reduction and internal fixation; anatomical reduction was achieved in 14 patients, acceptable reduction in 24 hips and unacceptable reduction in 1 hip; 24 hips developed AVN at the latest follow-up and the remaining 15 hips didn’t. Statistical analysis indicated that age ( t=3.875, P< 0.001), the severity of initial displacement ( Z=-2.118, P=0.034) and the rate of AVN ( χ2=42.280, P< 0.001) in patients with PPC were significantly higher than those in patients without; Logistic regression analysis confirmed age ( OR=1.288, P=0.011) and AVN ( OR=40.336, P< 0.001) as risk factors for PPC. ROC curve analysis indicated 10 years was the cut off age to significantly increase the rate of PPC. The rate of PPC in patients aged over 10 years (63.6%, 21/33) was significantly higher than that (24.7%, 18/73) in those aged less than 10 years ( χ2=14.848, P< 0.001). Conclusion:Age over 10 years and AVN are risk factors for PPC in children with femoral neck fractures treated surgically.

BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.

Objective To evaluate the effects of different interventios on insulin resistance and ?-cell function in population with impaired glucose tolerance(impaired glucose tolerance,IGT).Methods The total of 382 cases of IGT were divided into five groups according to interventions:control(C),diet plus exercise group(D+E),Acarbose Group(A),Metformin(M)and traditional Chinese medicine Group(TCM),with each group making the appropriate intervention programmes.BMI,FPG,2hPG,F-Ins,P-Ins,F-CP and P-CP were determined before intervention and yearly during the follow up,and BMI,HOMA-IR,HOMA-? were calculated.Results By the end of the study,insulin,F-CP,IR Group D+E and FPG,2hPG,insulin,C-peptide,IR in the other groups were significantly lower than before the intervention(P