Objective To observe the influence of intra-articular injection of lovastatin on cartilage degeneration and mRNA expres-sion of matrix metalloproteiase-1,-3 (MMP-1,-3) in cartilage and synovium of osteoarthritis (OA) model. Methods Thirty New Zeal-and rabbits underwent unilateral anterior cruciate ligament transection (ACLT) were random divided into two groups. The rabbits in experi-mental group received 0.1 mg/ml intra-articular lovastatin injection at 0.2 ml/kg of body weight immediately after surgery , once a week. Those in control group were treated with intra-articular injection of normal saline. At the 6th week after surgery, all rabbits were sacrificed. The articular cartilage degeneration of medial femoral condyle was evaluated under microscopy. Gene expression of MMP-1,-3 was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Cartilage degeneration in untreated group was significantly more severe than that in experimental group. In cartilage, no significant difference on the expression of MMP-1 and MMP-3 mRNA was found be-tween the experimental group and control group. In synovium, the expression of MMP-1 and MMP-3 mRNA was significantly suppressed in experimental group compared with control group. Conclusion During the development of experimental OA, intra-articular administration of lovastatin reduces the expression of matrix degrading enzyme, thus alleviate cartilage degradation.

Objective To explore morphologic characterizatics of cellular processes from adult human nucleus pulposus cells. Methods The nucleus pulposus of adult human intervertebral disc were obtained from 8 patients(Thompson's grade Ⅰ-Ⅱ) and then the tissues specimens were carried out by frozen section and electron microscopic section as well as cell isolation and cultured,processes of nucleus pulposus cells were examined using light microscopy,laser scanning confocal microscopy and transmission electron microscopy. Results When examined at both the confocal and electron microscope level,all the cells possessed the processes and adjacent nucleus pulposus cells processes possessed a gap junction.The elongated and round cells were examined when NP cells became monolayer in vitro.The rate of elongated cells to round cells was 2.3 to 1.The elongated cells protruded along with the long axis of cell body without second processes.Dendritic processes of round cells protruded to all directions from the cell body with multiple-level processes.Conclusion Processes are one of the morphologic characteristics of intervertebral disc cells.The research on processes functions could be helpful to understand pathomechanism of intervertebral disc degradation.

Objective To evaluate the effectiveness of thoracoscopic techniqu es for the anterior correction of adolescent idiopathic scoliosis. Methods Betwe en March 2000 and November 2002, 33 patients with adolescent idiopathic scoliosi s were selected to undergo thoracoscopic instrumentation, correction, and fusion . There were 26 females and 7 males. The average age at surgery was 13.6 years ( range, 10 to 18 years). All patients had idiopathic thoracic curves of typeⅠacc ording to the Lenke classification system, 24 of whom were typeⅠA, and 9 typeⅠ B. The average preoperative Cobb angle of the primary thoracic curve was 57.4?( range, 43? to 68?), and that of the compensatory lumbar curve was 32.0?(range , 20? to 47?). 27 patients were an averaged 6.3? of hypokyphosis (range, 0? to 18?). All 33 patients underwent endoscopic anterior release, instrumentation , correction and intervertebral fusion using the CD Horizon Eclipse system. Para meters including coronal Cobb angle of the primary thoracic curve, the compensat ory lumbar curve and thoracic kyphosis, were measured to assess coronal and sagi ttal correction. Results The instrumented levels were from T4 to T12. The averag e number of levels instrumented for each patient was 7.4 (range, 5 to 9). Operat ive times averaged 3 h and 48 min (range, 2 h 40 min to 5 h 30 min). Blood loss averaged 308 ml (range, 100 to 500 ml). Hospitalization averaged 4.4 days (range , 3 to 9 days). The average follow-up period was 20.6 months (range, 6 to 36 mon ths). The correction for the primary thoracic curve averaged 60.2%, and the comp ensatory lumbar curve averaged 50.3%. Correction with hypokyphosis averaged 20.4 ?. One patient developed pseudoarthrosis and rod fracture. Two patients experie nced transient chest wall numbness. Conclusion Anterior thoracoscopic correction of adolescent idiopathic scoliosis has the advantages of minimal invasion and s horter rehabilitation period. It can also provide the same corrective effect com paring to conventional open thoracotomy.

Objective To clone VEGF gene and construct its eukaryotic expression vector for the evaluation of the possability of VEGF gene therapy in orthopedic disease. Methods Human vascular endothelial growth factor(hVEGF) cDNA was amplified by nested PCR method from the HL60 cells and cloned to expression vector pcDNA 3. The cDNA was identified by enzyme digestion and DNA sequencing. Rabbit osteoblasts were transfered with pCD-hVEGF165 plasmid by lipofectin mediated gene transfer method. The transient expression of VEGF were detected by Streptavidin-Biotin-enzyme Complex(SABC). Results The cloned cDNA was confirmed to be VEGF165 cDNA. It was observed that the expression of human VEGF gene was detected distinctly 72 h after transfering. Conclusion We successfully cloned hVEGF 165 gene and construced its eukaryotic expression vector, which provided the further foundation of VEGF gene therapy for ostenecrosis,bone defeat and fracture.

The study assessed the early functional outcomes with cemented titanium implants of radius in the treatment of comminuted fractures of radial heads. The functional outcomes of arthroplasty with cemented titanium implants of radius in the treatment of radial head fractures (Mason Type III: 6; Mason Type IV: 4) in 10 consecutive patients (mean age, 38 years) were evaluated over a mean time of 23.7 months (18-31 months). The patients were assessed on the basis of physical examination, functional rating (Mayo) and radiographic findings. The parameters evaluated included motion, stability, pain, and grip strength. Five patients were considered to have excellent results, 4 patients had good results and 1 patient had fairly good results. There were no cases of infection, prosthetic failure, heterotopic ossification or dislocation. When medial collateral ligament was injured, radial head became the main stabilizing structure of the elbow. Titanium radial head implant may provide the stability similar to that of native radial head. We believe that titanium radial head implants may be indicated for the Mason Type III and Mason Type IV radial head fractures.
Elbow Joint/surgery ; Fracture Fixation/*methods ; Fractures, Comminuted/*surgery ; Prostheses and Implants ; Radius Fractures/*surgery ; Titanium ; Treatment Outcome

BACKGROUND: Allicin has been proved to influence the proliferation and apoptosis of LM-8 cells.OBJECTIVE: To study the effects of allicin on the expression of apoptosis protein Bcl-2 and Bax in C3H mouse osteosarcoma cell line LM-8, and the correlation of the morphological changes of LM-8 cells with the changes of Bcl-2 and Bax.DESIGN: Randomized grouping, non-blind, cell level, in vitro study.SETTING: The experiment was carded out from September 2006 to May 2007 at the Center Laboratory of Union Hospital,Tongji Medical College, Huazhong University of Science & Technology.MATERIALS: Osteosarcoma cell line LM-8 was purchased from the Animal Center of the Fourth Military Medical University of Chinese PLA. Allicin presented by Wuhan Huihai Co., was a sulphide isolated from garlic corn. Cell Counting Kit-8 (CCK-8) from Dojindo Laboratories; Bcl-2 antibody, Bax antibody and SP immunohistochemistry kit from Fuzhou Maixin Biotech Co.METHODS: LM-8 cells were cultured in RPMI 1640 culture medium, then cell climbing films were made, and SP immunocyte-histochemistry was used to detect the expression of Bcl-2 and Bax protein. Inverted microscope was performed to observe the morphologic changes of LM-8 cells before and after adding with 5.0, 10.0 and 15.0 mg/L allicin. Serial subcultivated LM-8 cells were adjusted to 7.5× 107 L-1 in concentration, added to 96-pore culture plate, and treated with allicin at a serial concentration of 1.0, 5.0, 10.0 and 15.0 mg/L. Additionally blank culture medium pores without cells and allicin and pores without allicin were taken as control groups. The 96-pore culture plate was maintained for 24, 48 and 72 hours, and then fetched. CCK-8 was added to determine the optical density value. The inhibition rate of LM-8 cell growth was also calculated. LM-8 cells were treated with different concentrations of allicin (5.0, 10.0 and 15.0 rag/L) for 24, 48 and72 hours, and then we carried out an apoptosis analysis by flow cytometry.MAIN OUTCOME MEASURES: Expression of Bcl-2 and Bax protein in LM-8 cells; cell morphous, cell proliferation,and cell apoptosis.RESULTS: ①Expression of Bcl-2 and Bax protein: Allicin could lower the expression of Bcl-2 protein and enhance the expression of Bax. The positive expression of Bcl-2, Bax and Bcl-2/Bax expression all showed significant differences compared with control group (P ＜ 0.01).②LM-8 cell morphous: LM-8 Cells treated with allicin presented typical apoptosis changes under microscope.③LM-8 cell proliferation: Allicin could inhibit the growth of osteosarcoma cell line LM-8, when concentration of allicin added from 1.0 mg/L to 15.0 mg/L, the inhibition rate of LM-8 cells at 72 hours increased from (23.87±3.26)% to (58.32±5.38)%, and 50% inhibiting concentration was 11.09 mg/L.④LM-8 cell apoptosis: After 72 hours with treatment of allicin of 5.0, 10.0 and 15.0 mg/L in concentration, apoptosis rate of LM-8 cells revealed a dose-dependent relationship (P ＜ 0.05).CONCLUSION: ①Allicin can inhibit the proliferation of LM-8 cells in a dose-time-dependent manner.②Allicin can induce the apoptosis of LM-8 cells in a dose-dependent manner.③Allicin can inhibit LM-8 cell proliferation and induce LM-8 cell apoptosis, which is related to down-regulate Bcl-2 protein expression and up-regulate Bax protein expression.

BACKGROUND: Thermosensitive hydrogel materials present stability at human body temperature, which is necessary for its application as a medical implant, thus the lower critical solution temperature (LCST) of thermosensitive hydrogel should be beyond the human body temperature by adjustment.OBJECTIVE: To prepare a thermosensitive hydrogel poly-N-isopropylacrylamide-co-N-hydroxymethylacrylamide P(NIPAAm-co-NHMPA) with over 37 ℃ LCST, and primarily appraise its safety as a medical implant in vivo.DESIGN: Random, non-blind, group control, animal experimental study.SETTING: Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology(HUST).MATERIALS: The experiments were carried out in the Central Laboratory, Union Hospital, Tongji Medical College, HUST and the Key Laboratory of Biomedical Polymer Materials of the Ministry of Education, College of Chemistry, Wuhan University between January and October in 2007. NIPAAm monomer and NHMPA monomer were purchased from Aldrich Company, crosslinking agent N, N'-methylene bisacrylamide from Fluka Company, and initiator ammonium persulfate and accelerating agent tetramethyl ethylene diamine from Sigma Company.METHODS: ①Taking ammonium persulfate and tetramethyl ethylene diamine as oxidation-reduction initiation system, while N, N'-methylene bisacrylamide as cross-linking agent, P-NIPAAm-co-NHMPA was prepared with the addition of NHMPA in the reaction system. LCST was determined by shrinking tests.②A series of biocompatibility tests such as sensitization test, acute systemic toxicity test, genetic toxicity test and implantation test were conducted in several experimental animals to evaluate the safety of the implant. MAIN OUTCOME MEASURES: The erythema and edema of stimulated lesions were recorded in sensitization test; the general state of each animal in acute systemic toxicity test were recorded 4, 24, 48 and 72 hours after injection; in genetic toxicity test, mouse bone marrow polychromatic erythrocyte (PEC) micronucleus was counted six hours after injection under microscope; sections after implantation were observed under light microscope.RESULTS: ①Synthesized hydrogel showed thermosensitive character and the LCST was 38 ℃.②In sensitization test, there was no erythema and edema occurred after leaching liquor and saline were injected; acute systemic toxicity test result revealed no symptom of toxicity; the genetic toxicity test suggested no difference of PEC frequency between experimental group and negative control group; in vivo implantation test, the inflammation around the material was mild and limited.CONCLUSION:P(NIPAAm-co-NHMPA) shows good biocompatibility and can be potentially used as an implant material.

AIM: To prepare and evaluate the biocompatibility of a thermosensitive hydrogel poly-(N-isopropylacrylamide-co-N-hydroxymethylacrylamide) (P-NIPAAm-co-NHMPA) in vivo so as to appraise its safety as a medical implant in clinical use. METHODS: The experiments were carried out in the Central Laboratory, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology(HUST) and the Key Laboratory of Biomedical Polymer Materials, Ministry of Education, College of Chemistry, Wuhan University between January and October in 2007. ① Preparation of thermosensitive hydrogel P-NIPAAm-co-NHMPA: Taking ammonium persulfate and tetramethyl ethylene diamine as oxidation-reduction initiation system, while N, N'-methylene bisacrylamide as cross-linking agent, P-NIPAAm-co-NHMPA was prepared with the addition of NHMPA in reaction system. Low critical solution temperature was determined by shrinking tests. ②A series of biocompatibility tests such as sensitization test, acute systemic toxicity test, genetic toxicity test and implantation test were conducted to evaluate the safety of the implant. RESULTS: ①Synthesized hydrogel showed thermosensitive character as expected, and the low critical solution temperature was 38 ℃.②In sensitization test, there was no sensitization proved in the material; in acute systemic toxicity test, the symptom of toxicity was not found; the genetic toxicity test suggested no teratogenic or mutagenic effects; in vivo implantation test showed that the inflammation around the material was mild and limited. CONCLUSION: Thermosensitive hydrogel P-NIPAAm-co-NHMPA has good biocompatibility and is a potential medical implant.

Objective To investigate the efficacy of transcutaneous injection of MIIGX3 artificial bone and methylprednisolone for recurrent bone cysts.Methods From January 2004 to March 2006,a total of 13 children with recurrent bone cysts received transcutaneous injection of methylprednisolone and MIIGX3 in our hospital.X-ray was employed to detect the degradation of MIIGX3 and formation of new bones.Results The operation time ranged from 30 to 95 minutes(mean 42 minutes).The patients were followed up for 1 to 3 years.None of them had recurrence of bone cysts during this period.The artificial bones were completely degraded and replaced by new bones in 1.5 years after the injection.Conclusions Transcutaneous injection of methylprednisolone and MIIGX3 is effective for recurrent bone cysts.Patients have shorter operation time and hospital stay after this treatment because the procedure is microinvasive.

OBJECTIVE:To explore the clinical efficacy and safety of Yunkang granule combined with dydrogesterone in the treatment of early threatened abortion. METHODS:128 patients with early threatened abortion were randomly divided into single drug group and combined medication group,64 cases in each group. Single drug group received Dydrogesterone tablet with first dose of 40 mg,then 10 mg,q 12 h until vaginal bleeding;combined medication group additionally received 8 g Yunkang granule, tid,until symptoms disappeared. Human chorionic gonadotropin (HCG),clinical symptoms remission time and disappearance time,pregnancy/childbirth complications,pregnancy outcome and neonatal adverse outcome,and the incidence of adverse reac-tions in 2 groups were compared. RESULTS:After treatment,HCG in 2 groups significantly increased,and combined medication group was higher than single drug group,the differences were statistically significant(P<0.05);clinical symptoms remission time and disappearance time in combined medication group were shorter than single drug group,the difference was statistically signifi-cant (P<0.05). There were no significant differences in incidence of pregnancy/childbirth complications,pregnancy outcome and neonatal adverse outcomes in 2 groups (P>0.05). The incidence of adverse reactions in combined medication group was 10.9%, which was significantly lower than single drug group (40.6%),the difference was statistically significant (P<0.05). CONCLU-SIONS:Both dydrogesterone alone and Yunkang granule combined with dydrogesterone show good efficacy in the treatment of ear-ly threatened abortion,but the remission time and disappearance time of clinical symptoms and adverse reactions in combined medi-cation group are less than that in single drug group.