Objective To investigate the effect of calcitonin gene-related peptide ( CGRP ) in inducing os-teogenic differentiation of rat precartilaginous stem cells in vitro and the underlying mechanisms. Methods Rat pre-cartilaginous stem cells ( PSCs) were cultured in complete osteogenesis medium containing DMEM/F-12 medium and different concentrations (0, 10-8,10-9,10-10mol/L) of CGRP, the morphology changes of PSCs were observed. The proliferation of PSCs was examined at different time points by CCK-8. All the PSCs were then randomly assigned to an experimental group and a control group. The PSCs in the experimental group were cultured in complete osteogenesis medium with 10-10 mol/L CGRP , while the control group cultured merely in complete osteogenesis medium was re-ceived no special intervention. Both groups were stained by Alizarin Red and the expression of alkaline phosphatase (ALP) was detected. The osteogenic genes (RUNX2,OPN and BGP) were measured by use of RT-PCR. The activa-tion of Wnt/β-catenin signaling pathway was tested by using Western blotting to evaluate the effect of CGRP . Results Compared to the control group ( the concentration of CGRP was 0 mol/L) , the concentration of ALP was significantly higher in the experimental group, calcium deposition was significantly more obvious, and the expression of the osteogenic genes such as RUNX2,OPN and BGP as well as theβ-catenin protein expression were up-regulated significantly. However, CGRP had no effect on cell proliferation. Conclusion CGRP activated Wnt/β-catenin sig-nal pathway and induced osteogenic differentiation of precartilaginous stem cells.

Objective To evaluate the safety and effectiveness of balloon kyphoplasty combined with physiotherapy in the treatment of osteoporotic vetcrbral compression fractures (OVCFs). Methods A retrospective review was of 12 OVCF cases ( including 16 fracture vertebrae) treated with balloon kyphoplasty was performed. Each patient had also been treated with anti-osteoporotic medication and the Rehabilitation of Osteoporosis Program-Exercise (ROPE) protocol, and each had received a six-month follow-up visit. The following parameters were recorded before the operation and 1 day and 6 months afterward : fracture recurrence, the severity of pain before and after treatment, and the change of vertebral height. The severity of pain was evaluated by using a visual analogue scale (VAS). Results During the treatment, no complication or adverse event was found. The average VAS values preoperation, postoperation and at the 6-month follow-up were 7.6±1.3, 2.1±1.2 and 2.6±1.4, respectively. The average changes in vertebral height preoperation, postoperation and at the 6-month follow-up were 49.2±18% , 74.3 ±14% and 69.8±16% respectively. All of the measures evaluated improved noticeably after the combined treatment. Conclusion Balloon kyphoplasty combined with anti-osteoporotie medication and physiotherapy showed good effects in the treatment of osteoporotic veterbral compression fractures.

ObjectiveTo explore the early effects of transplantation of fetal spinal cord (FSC) and basic fibroblast growth factor (bFGF) on calcium, magnesium of injured spinal cord.MethodsThe spinal cord injury (SCI) animal model was established with Wistar rats. The rats were divided into 3 groups. The group A was received FSC cells suspension 5 ml after injury. The group B was received FSC cells suspension and bFGF 20 μl. The group C was not treated with any therapy. The values of Ca2+, Mg2+ of injured spinal cord tissues were measured at 6, 24 hours after injury. The morphyological results were observed by electron microscope on the fifth day.ResultsThe degree of ionic disorder in FSC+bFGF treated rats was strikingly ameliorated and the contents of H2O also decreased. The morphyological finding revealed that in FSC+bFGF treated rats, the damage of gray and white matters was slight in contrast to those of control group rats, which appeared a typically damage.ConclusionTransplantation of FSC and bFGF has some protective effects on contused spinal cord in rats.

BACKGROUND: Multilevel cervical disc herniation accompanying ossification of posterior longitudinal ligament (OPLL) can seriously hurt the spinal cord. Which way of operative approach is more preferable is still uncertain. Whether one-stage combined anterior and posterior operation can achieve better effects remains unclear.OBJECTIVE: To explore the therapeutic effect of one-stage anterior and posterior operation for the treatment of multilevel cervical disc herniation accompanying OPLLMETHODS: Seventeen patients with multilevel cervical disc herniation and OPLL were treated by one-stage anterior and posterior operation at the Department of Orthopaedics, Tongji Hospital, were selected, including 11 males and 6 females, aged 42-74 years,mean aged 51.5 years. X-ray film, CT or MRI before operation showed the cervical cord was compressed by the multilevel herniated cervical discs or the ossified posterior longitudinal ligament. The stability and fusion of the injured segments were observed by regular postoperative X-ray film.RESULTS AND CONCLUSION: All patients were received a 6-36 months follow-up (24.5 months on average). The postoperative JOA scores were (12.88±2.47) points, which was greater than that of preoperatively [(6.41±1.28) points, P<0.05]. The improvement-rate of the spinal function after 6 months included 5 excellent cases, 7 good cases, and 4 fair cases. The excellent-good rate was 71%. All the patients got completely reduction, and all grafts got fused at 3-4 months after operation. The cervical intervertebral height and lordosis were satisfied maintained and there was no complication related to internal fixation breakage, loosening or displacement. It suggested that one-stage anterior and posterior operation can provide satisfied decompression earlier and rebuild the spinal stability in time, which is a safe and effective surgical intervention for multilevel cervical spondylotic myelopathy accompanying OPLL.

Objective: To investigate the differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities, and to screen for new candidate genes related to metastasis of osteosarcomas. Methods: The total RNAs of a lowly metastatic and a highly metastatic osteosarcoma cell lines (M6 and M8) were extracted. Differentially expressed genes in the two osteosarcoma cell lines were studied by cDNA microarray. The hybridization signals were scanned with a Generation Ⅲ array scanner and analyzed by Imagequant 5.0 software. Typical differentially expressed genes were further verified by real-time quantitative PCR. Results: There were 330 differentially expressed genes between M6 and M8 cells. In the high-metastasis M8 cells, 178 genes were up-regulated and 152 genes were down-regulated compared to the low-metastasis M6 cells, with 43 extremely up-regulated and 49 extremely down-regulated. The differentially expressed genes were mainly associated with cell proliferation, indicating these genes might be related to the inhibition of M6 cells. Other differentially expressed genes included those associated with the regulation of gene expression and signal transduction, indicating these genes might be correlated with tumor metastasis. Conclusion: cDNA microarray shows an advantage in identifying genes associated with metastasis of osteosarcoma. In M8 subset of MG63 osteosarcoma cells,43 genes are up-regulated and 49 genes are down-regulated, which may be related with metastasis of osteosarcoma.

Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells (PSCs). Methods Rats' PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro.The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode (LED) with an irradiation energy of 0.5 J/cm2,1 J/cm2,2 J/cm2 or 4 J/cm2.Any effect was confirmed by Alcian blue staining,immunohistochemistry and observing histomorphological changes under a light microscope,as well as detection using a reverse transcription polymerase chain reaction (RT-PCR). Results After being induced for 14 d,the PSCs exhibited polygonal and round shapes. Alcian blue and type Ⅱ collagen immunohistoehemistry staining showed positive results,but the control group had no significant change.RT-PCR showed that the mRNA expression of Sox9 and type Ⅱ collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.

BACKGROUND:Herba epimedi, a traditional Chinese medicine, has a long time in dealing with various orthopedic disorders. Icarinwithmany biological activites is one of the most important compositions of Herba epimedi. OBJECTIVE:Toinvestigate the effects of icarin on osteogenic differentiation of mesenchymal stem cels and the underlying mechanisms. METHODS:Bone marrow mesenchymal stem cels were treated using icarin with or without osteogenic mediumin vitro. Osteogenic differentiation markers, including runt-related transcription factor 2, osteocalcin and osterix, were detected by real time-qPCR. Alizarin red staining was used to measure calcium nodes generated by osteoblasts induced frombonemarrow mesenchymal stem cels. The proximal tibia bone structure of rats fed with icarin (2 mgperday) for 5 weeks was detected and analyzed by MicroCT. RESULTS AND CONCLUSION:Icarin was able to promote the expression of genes related to osteogenic differentiation in the absence or presence of osteogenic induction. Icarin could obviously increase the quantity of calcium nodes whenmesenchymal stem celswere cultured in the osteogenic medium. The animal experiment showed that icarin improved formation of trabecular bone.

Objective To investigate the minimal invasive management of cystitis glandularis with invasion of the ureteric meatus.Methods The clinical data of 18 cases were reviewed.Among the 18 cystitis glandularis patients,12 cases were invasion of bilateral ureteric meatus and 6 of unilateral ureteric meatus.Operation or drug treatment was performed on the diseases that can cause cystitis glandularis such as bladder stone,bladder neck stegnosis,external urethral meatus stegnosis and benign prostate hyperplasia.Sensitive antibiotic was administrated in all cases.After placing ureter catheter,transurethral plasma electro-resection was carried out in five patients whose ureteric meatus could be identified.In addition,of thirteen patients with ureteral orifice unable to be identified,there were ten cases with normal renal function,mitomycin was injected under affected membrana mucosa,and then the patient with ureteral orifice identified underwent transurethral plasma electro-resection after placing ureter catheter.On the other hand,the patient whose ureteral orifice still could not be recognized undertaken transurethral electro-resection at first,during which the ureter catheter was put once ureteral orifice had been detected,otherwise,the ureter catheter should be placed through cystoscope if nephritic colic emerged and hydronephrosis aggravated after operation.In those patients with kidney dysfunction,the ureter catheter was put by ureter discission or ureter replantation at first,the transurethral electro-resection could not be executed until the renal function recovered.Following all these procedure above,bladder instillation of drugs regularly,anti-infection and symptomatic treatment were administrated.Results One patient combined with bladder adenocarcinoma received cystectomy,of the other patients,six cases recurred and underwent electrotomy again resulting in no relapse.All nephrohydrops vanished or relieved obviously,nevertheless,urinary tract infection,haematuria and bladder-ureter backstreaming as the cardinal complication developed in some cases.Conclusion In the management of cystitis glandularis encroaching ureteric meatus,total or partial cystectomy can be avoided if ureter draining freely can be ensured,motivation removal,antiinfection,injection of drug under mucosa and preoperative diuresis conduce to the achievement of ureter catheter placing,transurethral plasma electro-resection is still effective methods in treating these cystitis glandularis.

BACKGROUND: Neuronal apoptosis produced a main effect in secondary injury after injury of central nervous system. Recently, minocycline has been reported to protect neurologic function evidently by decreasing apoptosis.OBJECTIVE:To study the effect of minocycline on apoptosis and expression of related factor, and protective effect of minocycline on neurologic function of injured spine after spinal cord injury (SCI) in rats.DESIGN: A randomized controlled animal study.SETTING: Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology.MATERIALS: The experiment was performed at the Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology between November 2003 and December 2004. A total of 36male SD rats were divided into four groups: ①Blank control group (group A) with 6 rats, ②injured group (group B) with 10 rats, ③minocycline treatment group (group C) with 10 rats, and ④methylprednisolone treatment group (group D) with 10 rats.METHODS: The animal SCI model was established. Spinal cords of rats in the group A were exposed without injury. Spines of rats in the group B were injured without treatment. Rats of group C was treated with minocycline (50 mg/kg) by intraperitoneal injection at 1 hour after injury, and 24 hours later another 50 mg/kg was given, and then another 25 mg/kg was injected for 5 days. Methylprednisolone (100 mg/kg) was intraperitoneally injected 1 hour after injury in the group D. The rats in the group A and the group B were treated with saline, and the method of administration was the same with that of the group C.MAIN OUTCOME MEASURES: ①Neurofunction evaluation of rats in each group. ②Expressions of Bcl-2 family members (Bcl-2, Bcl-xl, and Bax) were tested with immunocytochemical method. ③Apoptosis index of spinal tissue of rats of each group.RESULTS: A total of 36 rats were involved in the result analysis. ①Neurofunction of rats in the group C increased as compared with the group B (P ＜ 0.05), and there was insignificant difference between group C and the group D (P ＞ 0.05). ②Bcl-2 positive expressive cells in the group C was more than that in the group B, and showed obvious difference between the two groups [(62.53±7.76)%, (35.14±3.70)%,P ＜ 0.05]. There was insignificant difference between expressions of Bcl-xl and Bax in group C as compared with the group B (P ＞ 0.05). ③The number of apoptosis positive cells in the group B was more than that in the group C and D [(44.63± 5.90)%, (32.71± 5.26)%, (34.31± 6.84)%,P ＜ 0.05].CONCLUSION: Minocycline can up-regulate the expression of Bcl-2, and raise the ratio of Bcl-2/Bax, so as to inhibit the neural apoptosis of spine,which have protective effect on neurologic function of injured spine.

Objective To study the apoptosis of human colorectal cancer HT-29 cells induced by Aralia elata Seem leaf total saponin (ETS) and its effects on the expression of relevant proteins. Methods MTT assay was used to detect the proliferation of human colorectal cancer HT-29 cells cultivated with different concentrations (6.25, 12.5, 25, 50, 100, 200 mg/kg) of ETS. Hoechst33258 staining and laser confocal imaging were used to detect the apoptotic cells. Morphological changes were observed. The expressions of Bcl-2 and Bax were detected by immuno-histochemistry. Results ETS could induce apoptosis of HT-29 cells and apoptosis was in a dose-dependent manner in a certain range. ETS could decrease the expression of Bcl-2 and increase the expression of Bax in HT-29 cells (P<0.05, P<0.01), showing a significant dose-effect relationship. Conclusion ETS can induce the apoptosis of HT-29 cells, and the mechanism may be related to reducing the expression of Bcl-2 and increasing the expression of Bax.