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 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 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.

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.

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.

Objective To explore laboratory examination method of vaginal infection disease ,and to investigate the etiology of recurrent vaginitis .Methods 324 patients with recurrent vaginitis were selected to investigated the symptoms ,signs and vaginal discharge examination .Results (1)Among 324 patients ,293 cases (90 .4% ) of single infection and 31 cases (9 .6% ) mixed infec-tions were founded .There were 97 cases with VVC ,81 cases with CV ,17 cases with AV ,12 cases with BV ,and 14 cases with TV in the single infection cases .(2)In the 120 cases with VVC ,Candida albicans were accounted for 65 .0% ,non-Candida albicans strains were founded in 42 cases (35 .0% ) .(3)Among 28 cases of AV ,Escherichia coli ,Staphylococcus aureus ,Streptococcus aga-lactiae and Enterococcus f aecalis were main bacteria .Conclusion Recurrent vaginitis is common in clinical obstetrics and gynecolo-gy .Because of complex pathogeny ,clinicians should pay attention to laboratory results ,combined with the symptoms and signs , comprehensive evaluation ,improve disease diagnosis and treatment .

Objective To compare the clinical effects between continuous renal replacement therapy (CRRT) and intermittent haemodialysis (IHD) for the treatment of sepsis-induced acute kidney injury (AKI). Methods A prospective study was conducted. Seventy-three patients with sepsis-induced AKI admitted to the intensive care units (ICUs) of Tianjin Hospital and Tianjin First Center Hospital from January to December in 2014 were enrolled. They were randomly divided into two groups: CRRT group (n = 35) and IHD group (n = 38). Data were recorded for the patients in two groups before treatment, including acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, mean arterial pressure (MAP), urine volume, and the levels of C-reactive protein (CRP) and serum creatinine (SCr) before and 1 week after treatment, the time of recovery of urine volume, the length of ICU stay, the duration of organ support, and the incidence of cardiovascular events. Results There was no statistically significant difference in APACHE Ⅱ scores (21.63±2.46 vs. 21.34±2.46), MAP [mmHg (1 mmHg = 0.133 kPa): 71.26±10.70 vs. 75.74±15.17], urine volume (mL: 404.00±79.13 vs. 438.97±87.17), CRP (mg/L: 100.94±14.73 vs. 95.17±27.03), and SCr (μmol/L: 394.02± 50.26 vs. 390.47±54.42) before treatment between CRRT group and IHD group (all P > 0.05). One week after treatment, compared to the IHD group, CRRT could dramatically reduce the levels of CRP (mg/L: 41.05±10.15 vs. 60.21±14.78, t = 6.401, P < 0.001), SCr (μmol/L: 185.97±65.48 vs. 232.02±71.93, t = 2.862, P = 0.006), urine output recovery time (days: 7.94±3.06 vs. 11.08±3.71, t = 3.923, P < 0.001), the length of ICU stay (days: 9.54±3.39 vs. 13.42±3.89, t = 4.521, P < 0.001), organ support time (days: 3.23±2.70 vs. 6.34±3.36, t = 4.343, P < 0.001), and the incidence of cardiovascular events [23.53% (8/35) vs. 39.47% (15/38), χ2 = 5.509, P = 0.025]. Conclusion Compared to IHD, CRRT can more efficiently help patients with sepsis-induced AKI in removing excessive water, metabolic waste, and lower the levels of pro-inflammatory cytokines, maintain homeostasis of the internal environment, lower the adverse effects on cardiovascular system, so that it significantly improve the prognosis of patients, shorten the time of organ support and the length of ICU stay.

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 investigate the correlation between serum adiponectin level and severity of coronary artery disease.Methods We measured the serum concentrations of adiponectin in 135 patients whom had received CAG.The severity of coronary artery lesion was evaluated by Gensini score,the number of stenosis vessel and clinical manifestation.Results The serum concentrations of adiponectin in Gensini score 0 group(74.53?30.76 ng/mL)and score 40 group(28.96?11.87 ng/mL)(P40 group were lower than those in score 20-40 group(P