Objective To study the method of isolating and culturing stem cells from rat adipose tissue and to determine if adipose-derived stem cells(ADSCs)harvest from rat could differentiate into osteogenic and chondrogenic in vitro.Methods ADSCs were isolated from rat inguinal fat pads after extensive washing with phosphate-derived saline and digesting with collagenase.After primary culture in control medium and expanded to two passages,the cells were incubated in either an osteogenic medium or an chondrogenic medium for 2-4 weeks to induce osteogenesis and chondrogenesis,respectively.Evidences of Osteogenic differentiation,were detected by a ALP solution,and while chondrogenic differentiation was confirmed using the histologic Alcian blue staining at acidic pH.Expressions of osteocyte and chondrocyte specific genes were confirmed by RT-PCR.Results ADSCs can be isolated from rat adipose tissue and,expanded rapidly.It exhibited a heterogeneous population of fibroblast like cells morphologically.ADSCs induced to osteogenesis were stained positively for alkaline phosphatase activity after 2 weeks and formed mineralized nodular structures,as conformed by Von kossa staining.Expression of osteocyte specific genes,such as ALP,osteopontin,osteocalcin,was detected.ALP and osteopontin,were expressed constitutively in osteogenic medium after 2 and 4 weeks of culture.Expression of osteocalcin,was induced by osteogenic growth factors at 4 weeks.Induced to chondrogenesis cells were positive of Alcian blue staining under acidic conditions and expression of aggrecan and typeⅡ/Ⅹ collagen genes.Aggrecan and typeⅡcollagen genes were abundant after 2 weeks in chondrogenic medium.TypeⅩ collagen gene was detected at 4 weeks.Conclusion Adipose-derived stem cells can be isolated from rat adipose tissue.Their biological characteristics are similar with mesenchymal stem cells(MSCs),and have the potential to differentiate into osteogenic and chondrogenic lineage.It may be an idea source of ADSCs for tissue engineering.

[Objective]To evaluate the long-term clinical results of Herbert screw in the treatment of intra-articular fractures by retrospective analysis.[Method]From 1991 to 2006,fifty-five patients of different parts of intra-articular fracture treated with Herbert screw were followed up effectively,including scaphoid fracture group of 19 cases,the radial head fracture group of 25 cases,the femoral head fracture group of 11 cases.[Result]After an average follow-up time of 5 years and 11 months(ranged,1-13 years),the good-to-excellent rates were 94.7%,96%,72.7% according to the modified Mayo score,the Broberg-Morrey score and the Tompson-Epstein score.[Conclusion]The Herbert screw can provide strong fixation and stability in the long-term follow-up.It is an ideal material in treating intra-articular fractures.

From July 2006 to November 2008,16 patients with unstable intertrochanteric fractures were treated with cemented bipolar hemiprosthesis arthroplasty, including 5 males and 11 females, at a mean age of 84 years (range, 78-92 years) in the First Affiliated Hospital of Soochow University. Of them, 15 patients were followed with mean time of 14 months. The cemented bipolar hemiprosthesis was purchased from Beijing Montagne. The patients received 5-7 days of antibiotics following arthroplasty in addition to medicine of anticoagulation and analgesia. 1 case died of pulmonary heart disease 10 months after the operation and 3 cases had heterotopic ossification. There was no prosthesis loosening or dislocation. According to Harris hip scores, 3 cases were rated as excellent, 9 good, 3 fair, and 0 poor; the excellent and good rate was 80%. Results show that cemented bipolar hemiprosthesis arthroplasty is effective in treating elderly unstable intertrochanteric fractures, but operation indications require more attention.

BACKGROUND:At present, spleen tyrosine kinase is the new target of studying and treating rheumatoid arthritis. OBJECTIVE:To study the influence of smal molecule tyrosine kinase inhibitor HL131078 on the inflammatory cel infiltration and cartilage destruction of the knee joint of mice with col agen-induced arthritis. METHODS:Forty DBA/1 mice were randomly and evenly divided into blank, model, positive and experimental groups. Col agen type II (CII) solution and Freund’s complete adjuvant (including mycobacterium tuberculosis) were injected into the mice of the latter three groups through the tail to establish mouse models of col agen-induced arthritis. At 2 weeks after the the first immunization with CII, the mice in the positive group were intragastrical y given R406 (10 mg/kg), once a day, for 28 consecutive days. The mice in the experimental group were intragastrical y given HL131078 (10 mg/kg), once per day, for 28 consecutive days. RESULTS AND CONCLUSION:Compared with the model group, the mean arthritis indexes of mice in the experimental and positive groups started to decline at 29 and 26 days. In the experimental group, the cartilage destruction of mouse knee joint was obviously reduced and the inflammatory cel infiltration in the knee joints was obviously reduced, which was close to that in the positive group. The results demonstrate that the smal molecule tyrosine kinase inhibitor HL131078 can effectively reduce inflammatory cel infiltration and cartilage destruction in the knee joints of mice with col agen-induced arthritis.

Objective To investigate the preliminary results and complications of a tapered proximal femur modular stem in total hip arthroplasty (THA).Methods From October 2010 to December 2011,tapered proximal femur modular stems were used for THA in 50 patients (56 hips).There were 14males and 36 females,at a mean age of 61 years (range,25-82 years).Forty-four patients had unilateral THA and six bilateral THA.Hip osteoarthritis secondary to developmental dysplasia of the hip occurred in 15 patients,femoral neck fractures in 12,avascular necrosis of the femoral head in 10,primary hip osteoarthritis in nine,rheumatoid hip arthritis in two,malunion of femoral neck fracture in one,and femoral head fracture combined with posterior dislocation of the hip in one.The adopted femoral component was a tapered proximal femur modular stem.Femoral head-acetabulum interface composed metal-polyethylene in 34 hips,ceramics-polyethylene in 12 hips,and ceramics-ceramics in 10 hips.There were 48 hips with standard femoral head (28 mm) and eight hips with non-standard femoral head (＞28 mm).Results Mean period of follow-up was 11 months (range,6-19 months) and two patients (two hips) were lost to follow-up.Harris hip score improved from 36 points (range,4-71 points) preoperatively to 89 points (range,55-98 points) at the final follow-up.There was one patient with mild pain in the thigh,one moderate pain,but none severe or critically severe pain at the final follow-up.At the final follow-up,no migration or loosening of the implanted prostheses occurred; periprosthetic bone ingrowth fixation on the femoral side was achieved in 53 hips and fibrous stable fixation in one hip ; apart from one hip of ＜ 2 mm prosthetic subsidence,the remained revealed no subsidence of the prostheses.Intraoperative complications included acetabulum perforation in one hip and periprosthetic femoral fracture in one hip.Conclusion The short-term results are satisfactory,but the potential risk of fretting/corrosion and even breakage at the modular stem junction remains.

ObjectiveTo explore the therapeutic effect of selective antagonist-AM630 of cannabionid receptor 2 (CB2) in treatment of the titanium particles-induced inflammatory osteolysis.MethodsForty-five female BALB/c mice, 6-8 weeks old, were involved in the study, of which 15 mice were used as skull donors and the rest experimental animals were randomly divided into three groups, ie, black group, control group and treatment group, 10 mice per group.The mice model with air-pouch osteolysis induced by the titanium particles were established.The mice in the treatment group were injected with CB2 selective antagonist-AM630 (200 μg · kg-1 · d-1) intraperitoneally from two days before establishment of the air-pouch osteolysis model to two weeks after establishment of the model.Then, the mice were sacrificed and the pouch tissues were collected for molecular and histological analyses.The pouch membrane thickness and cell infiltration were tested by using computerized image analysis system and HE staining respectively.Osteoclast-like cells in the pouch membrane were determined by using tartrate-resistant acid phosphatase (TRAP) staining.Quantitative real-time polymerase chain reaction (RT-PCR) was employed to detect the mRNA levels of CB2, IL-1 β, TNF-α, receptor activator of NF-κB ligand (RANKL)and receptor activator of NF-κB (RANK).ResultsThere exhibited apparent erythematous and oedematous changes in the control group, which was mitigated around the bone implants with AM630 treatment.Quantitative image analysis of the histological sections revealed significant difference of the pouch membrane thickness among three groups, (192.2 ± 19.4)μm in control group, (88.5 ± 14.7) μm in blank group and (122.1 ± 15.2) μm in treatment group (F = 101.74, P ＜ 0.05).Intensive TRAP staining was identified much in the control group but markedly reduced after AM630 treatment in the pouch tissues.RT-PCR showed that titanium particle stimulation could enhance the expressions of CB2, IL-1 β, TNF-α, RANKL and RANK gene in the air pouch tissues.However, the mRNA levels of these genes were markedly reduced after AM630 treatment, with statistical difference compared with control group (P ＜ 0.05).ConclusionsCB2 selective antagonist AM630 can inhibit the process of titanium particlesstimulated inflammatory reaction and osteoclast activation.Therefore, CB2 represents a new suitable therapeutic candidate for the prevention and treatment of aseptic loosening of the artificial joint.

BACKGROUND:There are a variety of surgical approaches for minimally invasive total hip replacement,such as anterolateral,posterolateral and two-incision.However,it is unknown which one is more in line with the concept of minimally invasive and easy to carry out.OBJECTIVE:To observe the clinical application effect of total hip replacement using anterlateral modified Watson-Jones approach.METHODS:From January 2005 to December 2006,35 patients were performed total hip replacement with the new anterlateral approach at the Department of Orthopaedics,the First Affiliated Hospital of Soochow University,including 16 males and 19 females,aged 52-78 years,mean aged 65.9 years;the average body mass index was 24.4 kg/m~2 (19.3-30.1 kg/m~2).The length of incision,operative duration,transfusion,complications,postoperative movement,postoperative hospital stay,prosthesis position and Harris score after operation were observed.RESULTS AND CONCLUSION:The patients were followed up 6-24 months,with an average of 17.3 months.The average length of incision was 8.3 cm (7-12 cm).The mean operative duration was 92.1 minutes (70-120 minutes).The average blood transfusion was 300 mL (0-800 mL).All the patients were practice their limb muscles immediately after the operation and they were activity the day after operation under the guidance of doctor.The average postoperative hospital stay was 7.9 days (5-15days).Eleven hip pains occurred at 3 months,9 cases at 6 months and 5 cases at 12 months,the Harris scores were (79.5±4.4)points,(84.0±3.6) points,and (85.4±3.0) points,respectively.No infection,dislocation,vessel or nerve injury happened.From the follow up,all of the patients had good prosthesis position and rang of hip motion.The results revealed that the new anterolateral approach is minimally invasive total hip replacement,and the patients can recover faster via some proper exercise.

Objective To compare the outcome of proximal femoral nail antirotation (PFNA)and third generation of Gamma nail (TGN) in the treatment of femoral intertrochanteric fracture in the elderly. Methods From January 2007 to June 2008, the patients with intertrochanteric fractures were divided into PFNA group ( n = 55 ) and TGN group ( n = 52 ) randomly. The two treatment groups were comparable in aspect of general data. The clinical data of surgical trauma, complications and postoperative function of the two groups were compared in our study. Results The mean operation time was (66.6 ± 15.4) min in PFNA group and (73. 1 ± 20.8 ) min in TGN group (P ＞ 0. 05 ). The mean intraoperative blood loss differed significantly, (219.5 ± 107.5 ) ml in PFNA group compared with (269.0 ±123.9) ml in TGN group ( P ＜ 0. 05 ). The mean fluoroscopy time was ( 2.97 ± 1.14 ) min in PFNA group and (3.43 ± 1.18) min in TGN group (P＜0.05). Unilateral femoral shaft splits at the distal end of the nail was observed intra-operatively in four patients of the PFNA group and in one of the TGN group,with no statistical difference between two groups. Of all, 91 patients ( n =46 in the PFNA group and n =45 in the TGN group) were followed up for a duration of ( 17.5 ±3.69) months. There was one re-fracture and one implant failure during the follow up period in PFNA group, which was not found in TGN group. Fiftyone patients recovered to normal activity status, including 27 in the PFNA group and 24 in the TGN group,with no statistical differences. Conclusions Intertrochanteric fracture can be treated successfully with PFNA and TGN. There is no significant difference in complications and functional outcome between the two groups. However, PFNA takes advantages of less blood loss, less operative time and less fluoroscopy time over TGN.

BACKGROUND:There are many methods to treat thoracolumbar fractures However,it remains unclear whether a simple operation to treat a thoracolumbar burst fracture can decrease the trauma and obtain better effect to avoid the enlargement of operation.OBJECTIVE:To investigate the clinical outcome of posterior short-segmental transpedicular screw system internal fixation in treatment of single-level thoracolumbar burst fracture without nerologic deficit.METHODS:A total of 186 patients with single.level thoracolumbar burst fracture but without nerologic deficit treated in the First Affiliated Hospital of Soochow University between September 2003 and January 2008,including 152 males and 34 females,aged18 10 65 years.were treated by posterior pedicle screw The intraspinal bone fragments were treated with indirect decompression.All patients were checked with radiography and CT scan before/after operation as well as before implant removal.The correction of anterior vertebral body height and the ratio of bone fragment to cross section area of spinal canal were measured.RESULT AND CONCLUSION:Compared with normal.the anterior vertebral body height was 42%before operation,98%after operation,and 98%before implant removal The ratio of bone fragment to cross section area of spinal canal was 34%before operation,13%after operation.and 8%before implant removal.Internal posterior short-segmental transpedicular screw fixation treating single-level thoracolumbar burst fracture without nerologic deficit can obtain stability of spinal column,and it is beneficial to recovery of body height,physiologic postu ral contour and canal volumn .

BACKGROUND:Studiesin vitro have suggested that icarin can attenuate lipopolysaccharide (LPS)-induced acute pneumonia. Is the anti-inflammatory effect of icarin stil valid in the presence of wear particles? OBJECTIVE:With studiesin vivo andin vitro, to investigate the regulatory effect of icarrin on titanium particle-induced inflammatory reaction. METHODS:(1) Studiesin vivo: Eighty male C57BL/6 mice aged 6-8 weeks were randomly divided into four groups: control group, icarin group, titanium particle group, and titanium particle+icarin group. Mice in the titanium particle group and titanium particle+icarin group received surgical procedure, and sterile and endotoxin-free titanium particles were implanted on the calvaria surfaces to induce inflammatory reaction. Mice in the control group and icarin group received the same surgery, but no wear particles were implanted. Then icarin was given oraly to mice in the titanium particle group and titanium particle+ icarin group with a dose of 200 mg/kg per day for 2 weeks from the day of modeling. Mice in the control group and icarin group were given oraly the same dose of placebo. Two weeks later, tumor necrosis factor-α and interleukin-1β at protein and mRNA levels were respectively detected with enzyme-linked immunohistochemical (ELISA) and quantitative real time reverse transcription PCR (qRT-PCR) analysis. (2) Studiesin vitro: Mouse monocyte/macrophage RAW264.7 cels were cultured with different conditioned media: control group, nuclear factor receptor ligand кB (RANKL); icarin group, RANKL+icarin; titanium particle group, RANKL+titanium particles; titanium particle+icarrin group, RANKL+icarin+titanium particles. Titanium particles stimulated RAW264.7 cels were co-cultured with RANKL and icarin for 72 hours. Tumor necrosis factor-α and interleukin-1β at protein and mRNA levels in the supernatant were detected with ELISA analysis and qRT-PCR, respectively. RESULTS AND CONCLUSION: (1) Resultsin vivo: icarin treatment obviously decreased titanium particle-induced inflammatory cellinfiltration and made the thickness of periosteum thinner, down-regulated tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels. (2) Results in vitro: when RAW264.7 cels were stimulated with titanium particles for 72 hours, tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels in culture media increased obviously; when icarin was administrated, tumor necrosis factor-α and interleukin-1βexpressions at protein and mRNA levels down-regulated significantly. These results suggest icarin can obviously suppress titanium particle-induced inflammatory reactionin vivo andin vitro.