BACKGROUND: Injectable hydrogel-chitosan is the latest developed material for cartilage tissue engineering. It provides a novel cell attachment mechanism. OBJECTIVE: To investigate the effect of allologous chondrocytes in combination with chitosan in repairing cartilage defects. DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at the Central Laboratory of China Medical University from November 2004 to March 2006. MATERIALS: Thirty 2-week-old New Zealand rabbits weighing 0.3 kg were selected to model cartilage defect. METHODS: Thirty rabbits were randomly divided into three groups. In the experimental group, the defects were repaired by the injection of second passage of chondrocytes (2.5?1011 L-1) combined with chitosan into articular capsule; the cell suspension group was only injected with chondrocyte suspension (2.5?1011 L-1); the control group was not given any treatment. MAIN OUTCOME MEASURES: The animals were sacrificed at the 4th, 8th, and 12th weeks after operation, respectively to harvest the lateral condyle of knee joint. Hematine-eosine staining, gross and histological examinations, and electron microscopic evaluations were performed to observe the repair condition. RESULTS: The gross observation showed that there was no obvious swelling or other inflammatory responses in each rabbits. Histological examination suggested that in the experimental group, the cartilage defects were repaired by the hyaline cartilage tissue with smooth surface, which integrated smoothly with articular cartilage at 12 weeks after operation. No obvious defect repair was observed in the other groups. The transmission electron microscopic observation showed that in the new-formation tissues of the experimental group, there were round or oval chondrocytes, and abundant rough endoplasmic reticulum, ribosome and Golgi complex in cytoplasm. At 8 to 12 weeks, plenty of collagen fibers were found in cell suspension group. In the control group, only fibrous tissues were seen on the surface of defects. CONCLUSION: The articular cartilage defects of rabbits are repaired with allograft of chondrocytes seeded onto chitosan, indicating chitosan can serve as carrier in cell transplantation.

Objective To explore the influence of abnormal mechanical stress on acetabular development, especially on the chondrocyte proliferation in acetabular growth plate, and to research the methods of the repair of acetabular dysplasia. Methods 60 Wistar rats of 3 weeks old were divided into three groups, and each group had twenty rats. The left side was experimental side, and the right side was control. In group A, the hip joint was dislocated by manipulation repeatedly within two weeks; in group B, the model of acetabular dysplasia was created by keeping the knee joint in extension through steel needle fixation. The needle was removed after two weeks of fixation; and in group C, the knee joint was fixed in continuous extension with steel needle. When at 5, 7, 9 and 12 weeks old, the rat acetabulum was observed through the soft X-ray photograph, histological method and electronic micrograph respectively. Results In group A, when the rat was 5 weeks old, the acetabular angle was larger roughly 5? than the control side. Malalignment of chondrocyte column in proliferative zone was observed. No difference was seen between the two sides at 7, 9 and 12 weeks old. In group B, when the rats was 5 weeks old, the acetabular angle increased compared to the control side. When the rat was 7 weeks old, malalignment of the chondrocyte column in proliferative and hypertrophic zone increased. When at 9 and 12 weeks old, histological changes became indistinct gradually. In group C, the acetabular angle kept increasing to the control side. The acetabular dysplasia became obvious and had no improving tendency. When the rat was 12 weeks old, the acetabular edge became adducting and flat. There was no obvious alignment of chondrocyte column. The nucleus of acetabular chondrocyte of the proliferative zone became smaller, endoplasmic reticulum and mitochondria decreased, and bubbles formed. Conclusion Acetabular dysplasia could recover when abnormal mechanical stress was released, and the congruence of the head and acetabulum were corrected in the highly growing period of acetabulum. The crucial reason to acetabular dysplasia is the metabolic changes of chondrocytes in proliferative zone of growth plate, which leads to tardy ossification of the acetabulum.

Objective:To investigate the interfering effect of siRNA on the expression of VEGF in Hela cells.Methods:Three pairs of siRNA were designed according to the 1-5 extrons sequence of VEGF and synthesized by T7 RNA ploymerase transcription in vitro. To evaluate the inhibition activity of siRNA, Hela cells were transfected via siPORT Lipid. The interfering effect of siRNAs in hRPE cells was examined by semi-quantitative RT-PCR and immunofluorescence technique.Results:The results showed that the three pairs of siRNA could effectively inhibit gene expression of VEGF in Hela cells with the rates of 88.7%, 94.2% and 80.3%. But the 1-base mismatched siRNA and ssRNA(+) didn′t show significant interfering effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect(5-10 pmol).Conclusion:The siRNAs synthesized by T7 RNA ploymerase in vitro could effectively and specifically interfere the expression of VEGF in Hela cells, providing a novel approach for gene therapy of tumor.

Objective To explore the capsular changes at the weight- bearing area of hip with osteoarthritis and its correlation to postoperative clinical results of Chiari pelvic osteotomy. Methods According to the Bombelli radiological classification, the 66 hips with osteoarthritis were classified into three groups: hypertrophic, normotrophic and atrophic. Specimens of the capsule at weight- bearing area were obtained during total hip replacement for osteoarthritis, and stained with HE, Van- Gieson and Safranin O. Histopathological changes of the capsules were observed and compared among the three groups. Results The hypertrophic group showed a high incidence of metaplastic cartilage (63.3％ ). Normotrophic group(32.1％ )and atrophic group(25.0％ ) showed a low incidence of metaplastic cartilage. The atrophic group had a higher incidence of fibrous synovitis or fibrosis(81.3％ ) than hypertrophic. Among the three group, the frequency of infiltration granulation tissues was low, the frequency of both infiltration of inflammatory cells and degeneration of collagen fiber was high. Conclusion Associating with clinical research, these findings suggest that the capsular changes at weight- bearing area are responsible for postoperative clinical results of Chiari pelvic osteotomy. Compared with normotrophic and atrophic type, the hypertrophic type has the best adaptation of Chiari pelvic osteotomy.

Objective To study the clinical characters and the prognosis of the sciatic nerve injury associated with acetabular fracture. Methods From January 1980 to January 2000, fourteen sciatic nerve injury resulted from acetabular fracture were observed. Three were treated with skeletal traction, eleven were treated with open reduction and internal fixation and neurolysis. Results All cases were followed-up for an average of 18 months. The anatomical reduction of acetabular fracture obtained in 57.1％ of the patients and satisfactory reduction in 42.9％ . The excellent and good nerve functional recovery was seen in 57.1％ according to MCRR. Conclusion Injury of the sciatic nerve associated with the acetabular fracture occurs most commonly when the hip is dislocated posteriorly. Injury of the common peroneal nerve, branching off from the sciatic nerve was more common and had poorer prognosis than that of the tibial nerve. Flexion of the knee and extension of the hip during the surgical reduction of acetabular fracture can reduce iatrogenic injury of the sciatic nerve. Early reduction of acetabular fracture and neurolysis is important for the recovery of function of the sciatic never.

BACKGROUND:Lipopexia induced by glucocorticoid is fat precipitation in marrow due to abnormal lipomatabolism,or differentiation of cells resulting from hormone-affected bone marrow mesenchymal cells.The precise generating procedure remains unclear.OBJECTIVE:To Jnvastigate effects of vadous concentrations of dexamethasone on adipogenic differentiation of bone marrow mesenchymal cells.DESIGN,TIME AND SETTING:The observational study was performed at the Department of Biochemistry,China Medical University from January 2007 to January 2009.MATERIALS:A total of 80 Sprague-Dawley rats aged 3 or 4 weeks,weighing (110±10) g,of both genders,were used in this study.METHODS:Rat bone marrow mesenchymal cells were isolated and subcultured.Bone marrow masenchymal cells at the third passage were used as samples.MAIN OUTCOME MEASURES:Cellular morphologic changes after treatment of 10~(-7) mol/L dexamethasone were observed under an inverted microscope,as well as at 3,7,14 and 21 days under normal culture condition.Morphological changes of bone marrow mesenchymal cells were observed following alkaline phosphatase and Sudan Ⅲ staining.Alkaline phosphatase activity was measured using phenol reagent.Effects of dexamethasone on cell proliferation were measured using MTT assay.RESULTS:Following 24 hours of incubation,a few adherent cells were found in the bottom of the culture flask,showing spindle shape.With prolonged time,adherent cells became more,presenting radiated shape.Following passage,cells distributed uniformly,showing typical fibroblast-shape.Following dexamethasone stimulation,cells changed from spindle-shape into polygonal or irregular shape.In the late phase,with increased concentration and prolonged time,cell colonies disappeared;cells adhered,and died.In normal cultured cells,no or a few orange particles were found.In dexamethasone-cultured cells,with increased hormone concentration and prolonged time,Sundan Ⅲ stained particles increased.At 21 days,alkaline phosphatase activity under normal culture was separately 1.57-,4.49-and 5.0-fold of 10~(-8),10~(-7),10~(-6) mol/L dexamethasone group.At 7,14 and 21 days,alkaline phosphatase activity under normal culture was separately 2.93-,3.80-and 4.39-fold of 10~(-7) mol/L dexamethasone group (P ＜ 0.05).High concentration of dexamethasone had significant inhibitory effects on cell proliferation activity.Significant difference was detected as compared 10~(-7) mol/L and 10~(-6) mol/L to other groups (P ＜ 0.05).CONCLUSION:High-dose dexamethasone enhances adipogenic and suppresses osteoblastic differentiation of bone marrow mesenchymal cells.The effect will increase along with the increasing content and prolonged duration of demamethasone stimulation.

Objective: To clone and express the recombinant human Vasostatin120-180aa domain and to investigate its activity of inhibiting angiogenesis in CAM. Methods: After amplifying gene of human Vasostatin120-180aa domain, we sub-cloned it into pQE30 vector and expressed Vasostatin120-180aa domain by E. coli. We also tested its ability of inhibiting angiogenesis in CAM. Results: The total gene length of human Vasostatin120-180aa domain is 180 bp. Expressed by pQE30 system in E. coli and purified by IMAC, Vasostatin120-180aa was detected by SDS-PAGE, in which there is a positive band and molecular weight is about 8 kD. Conclusions: Recombinant human Vasostatin120-180aa could play effective role in anti-angiogenesis in CAM and it showed a dose dependent effect in some degree.

Objective To explore the prevalence,subtype,distribution characteristics and risk factors of IBS met ROME Ⅲ criteria in local adult residents of Shanghai Songjiang area.Methods With multi-stage,stratified cluster random sampling method,from April to May in 2010,residents of45 shanghai Songjiang communities were interviewed by questionnaire survey.IBS diagnosis was based on Rome Ⅲ criteria,and the subtypes were according to Bristol stool scale.Self-rating anxiety scale (SAS),self-rating depression scale (SDS) and Athens insomnia scale (AIS) were also simultaneously finished. Results There were 7648 individuals who completed the survey with qualified questionnaires; the efficient rate was 90％. The sample size accounted about 1.62％ of total population in Shanghai Songjiang area.Total 970 patients were detected,who met the Rome Ⅲ criteria,of those 492 males and 478 females.The adjusted prevalence of IBS was 13.1％,13.8％ in males and 12.7％ in females.The ratio of male to female was 1 ∶ 0.92.There was no significant difference in detection rate between males and females (P＞0.05).The prevalence was highest in the 18 to 29 years age group (18.2％,P=0.000).IBS unsubtyped (IBS-U) was the most common type in patients who met the Rome Ⅲ criteria (45.3 ％),IBS with diarrhea (IBS-D) type was more common in male (27.0％) and IBS with constipation (IBS-C) was more common in female (19.5％).The IBS prevalence of obesity group (x2 =4.046,P=0.044),higher education group (x2=31.210,P=0.000),mental group (x2 =8.409,P =0.015),unmarried group(x2 =26.933,P =0.000) and no abdominal surgery history group(x2 =5.894,P=0.015) was higher than that of control group.The risk of IBS prevalence in those who had the history of gastrointestinal infection,abdominal surgery,taking antibiotics or analgesics increased by 5.105,3.388,2.949 and 2.811 times respectively(P=0.000).Multivariate analysis indicated that the risk of IBS prevalence was increased by 1.69 times if took more spicy food,however high-protein diet was a protective factor (OR=0.900,P=0.000).Anxiety (OR=2.452,P=0.000) and insomnia (OR=1.923,P=0.000) were also risk factors of IBS.Conclusion IBS is a common and frequently occurring disease in Shanghai Songjiang community.The history of gastrointestinal infection,abdominal surgery,taking antibiotics or analgesics,intake of more spicy food,anxiety and insomnia may be risk factors of IBS.

Objective To investigate the role of additional post core biopsy ultrasound in clinically node negative breast cancer.Methods Axillary ultrasound was performed before and after breast cancer was diagnosed on core biopsy samples.Post core biopsy ultrasound were performed by radiologists of this department of ultrasound at random.Post-diagnosis ultrasounds were performed by a radiologist with over 20 years of experience for the diagnosis of breast cancer with axi[lary disease.Results were compared to the final axillary pathological result.Results Of the 96 patients,17 were pathology lymph node positive.Post biopsy ultrasound identified 8 of the 17 positive nodes,with a sensitivity 47.1％,specificity 88.6％,positive predictive value of 47.1％,negative predictive value of 88.6％,accuracy of 81.3％.While the diagnosis index of pre-biopsy ultrasound were 47.1 ％,88.6％,47.1％,88.6％,81.3％,respectively.Conclusions Post-biopsy ultrasounds had an increased sensitivity for identifying positive axillary nodes,at the same times,specificity decreased.

Objective To analyze the clinical features,methods of treatment and prognosis of primary neurogenic tumors of mediastinum in patients taking surgical intervention.Methods A database was maintained retrospectively of all patients undergoing surgery for tumor and pathologically diagnosed with primary neurogenic tumors of mediastinum,managed in the Department of Thoracic Surgery,Zhongshan Hospital,Fudan University,Shanghai between Jan.,2008 and Dec.,2014.This work analyzed retrospectively the information about clinical and imaging features,surgical techniques and outcome extracted from medical records.Results Among the 131 cases,78 cases (59.5％) were males,53 cases (40.5％) were females;72 cases were diagnosed incidentally (55.0％),while the other 59 cases (45.0 ％) suffered from different symptoms.The posterior mediastinum was the most principal location with 61 cases in the left and 69 cases in the right,and 1 case remained in the anterior mediastinum.Total 98 cases (74.8％) underwent surgeries via video-assisted thoracic surgery (VATS),5 cases (3.8％) took VATS surgery with small incision,and 28 cases (21.4％) experienced open thoracotomy,with no mortality during perioperative period.Gross total resection was obtained in 130 patients (99.2％).The remaining patient underwent a palliative resection for malignant schwannomas.Of the patients,98 cases had benign schwannomas (74.8％),24 cases had gangliocytomas (18.3％),2 cases had malignant schwannomas (1.5％),2 cases had neurofibromas (1.5％),2 cases had paragangliomas (1.5％),2 cases hadprimitive neurotodermal tumor (PNET) (1.5％) and 1 case had neuroblastomas (0.8％).All patients were followed up from 12 to 95 months with an average of 53 months.A patient with PNET died of tumor metastasis,a patient with malignant schwannomas died after palliative ectomy,and 2 cases died of other reasons.The rest survived until Jan.,2016 with tumor free.Conclusions Nearly no specific clinical symptoms occur in neurogenic tumors of mediastinum.Most of neurogenic tumors of mediastinum are benign with optimistic prognosis after surgical treatment.While malignant neurogenic tumorsusually come with poor prognosis,which places special emphasis on early diagnose together with surgical treatment.