Objective To investigate the chondrocyte proliferation and extracellular matrix biosynthesis of electrospun PCL scaffolds seeded with rabbit chondrocytes under flow perfusion culture in vitro.Methods Nonwoven PCL microfiber mats were fabricated,and contra-aperture cylindrical glass equipment as a perfusion bioreactor was designed and manufactured on our own.The experiment included peffusion culture group and static culture group.Primary chondrocytes were isolated from the knee joints of two-month-old New Zealand white rabbits and seeded into scaffolds.The scaffold-cell complexes were harvested at 3,7,and 14 days of culture for scanning electron micrograph (SEM) analysis,biochemical assay,real-time PCR and histology analysis.Results Electrospun PCL scaffolds were composed of microfibers with a diameter of 1.67±0.76 μm and pores with a diameter of 17.65±7.11 μm.SEM showed a better cell proliferation with typical morphology of chondrocytes under perfusion culture.At 7 days of culture,DNA content in perfusion culture group was higher than in static culture group.At 3,7 and 14 days of culture,compared with the static culture group,glycosaminoglycan (GAG) content and GAG/DNA ratio in perfusion culture group were higher,and the differences were statistically significant.At 14 days of culture,real-time PCR showed aggrecan and collagen type Ⅱ gene expression and collagen type Ⅱ to collagen type Ⅰ ratio were higher in perfusion culture group than in static culture group; HE and safranin O staining showed a significant cell proliferation,infiltration,as well as extracellular matrix biosynthesis in perfusion culture group.Conclusion Under flow perfusion culture,the electrospun PCL scaffolds seeded with rabbit chondrocytes can enhance chondrocyte proliferation and extracellular matrix biosynthesis,which is a promising method for cartilage tissue engineering.

BACKGROUND: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, proliferation. Currently, no perfect scaffold is available. OBJECTIVE: To observe the growth of rabbit bone marrow stromal cells (BMSCs) cultured in different-intensity three-dimensional fibrin glue in vitro, and to discuss the feasibility of fibrin glue used as a scaffold material of bone tissue engineering. DESIGN, TIME AND SETTING: The single sample observational study was performed at the China-Japan Union Hospital of Jilin University and School of Mechanical Engineering of Tianjin University of Technology from September 2007 to January 2008. MATERIALS: Fibrinogen and thrombin were mixed at various proportions, and prepared into different intensity fibrin glue. A month-old male New Zealand white rabbits, weighing 0.25 kg was used in this study. METHODS: Rabbit BMSCs were cultured and serial subcultivation in a CO2 incubator. And then the amplified BMSCs were collected and continue to be cultured in different intensity fibrin glue for 4 weeks. MAIN OUTCOME MEASURES：Observation of growing BMSCs is performed using the phase contrast microscope. The activity of BMSCs in fibrin glue at different stages was observed using hematoxylin-eosin staining. The ultrastructural changes of BMSCs were observed which had been cultivated in fibrin glue for 4 weeks. RESULTS: After growing in fibrin glue for 4 weeks, BMSCs showed strongly active status in low intensity fibrin glue and growing slowly or dying in high intensity fibrin glue. Under the electron microscope, BMSCs following 4 weeks culture in fibrin glue (proportation of fibrinogen and thrombin was 4:1) were found, with visible cellular organs, and BMSCs had good activities. CONCLUSION: BMSCs can spread and proliferate quickly in low intensity fibrin glue. The optimal proportion of fibrinogen and thrombin is 4: 1.

BACKGROUND: A superior composite scaffold hopes be constructed to resolve adhesion between seed cell and scaffoldmaterial.OBJECTIVE: To construct composite scaffolds with fibrin glue and xenogeneic inorganic bone and to explore the three-dimensional culture of rabbit marrow stromal cells (MSCs).DESIGN, TIME AND SETTING: Observational experiment was performed at the Department of Orthopedics of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University from November 2007 to March 2008.MATERIALS: Fibrin glue was made by a certain ratio of fibrinogen and thrombin; bovine cancellous bone following defatting and deproteinization was mixed with fibrin glue to establish composite scaffold.METHODS: Rabbit MSCs were cultured in v#ro and transferred, and the MSCs were collected for three-dimensional culture withcombined scaffolds made of xenogeneic inorganic bone and fibrin glue.MAIN OUTCOME MEASURES: The growth and proliferation of MSCs were examined by phase-contrast microscope andtransmission electron microscopy.RESULTS: Phase contrast microscope showed that the MSCs could spread evenly in the combined scaffolds. After cultured 4 weeks, the MSCs formed into densely three-dimensional net. It could be observed under the transmission electron microscopethat there were micro-protrusions in local stromal cells at 4 weeks after culture, and the mitochondrion as well as ribosomes wasobserved in the cytoplasm with rough endoplasmic reticulum.CONCLUSION: The MSCs cultured in the combination of fibrin glue and xenogeneic inorganic bone scaffolds show a betteractivity, and they can proliferate rapidly.

ObjectiveTo explore the effect of osteoarthritis (OA) chondrocytes and normal chondrocytes on the chondrogenesis of bone marrow mesenchymal stem cells(BMSCs) in self designed co-culture system. MethodsRabbit BMSCs and chondrocytes were isolated and expanded in vitro. OA chondrocytes were harvested from the rabbit of established osteoarthritis model. We made a BMSCs-low melting agarose constructs, then put it onto the self-made 6 well plates lattice assembly for co-culture with chondrocytes. The groups were divided into Normal P0-BMSCs, Normal P3-BMSCs, OA PO-BMSCs, OA P3-BMSCs and BMSCs (control) group. At 3, 7, 14 day culture, cultured cells in all groups were collected for real-time PCR, glycosaminoglycan (GAG) content, cytoactive detection, and histological observation. ResultsType Ⅱ collagen gene expression was up-regulated in group Normal PO-BMSCs, which showed 5.1-, 7.2-, 11.2-fold increase over that of control group at 3, 7, 14 days, respectively. Type Ⅰ, Ⅱ collagen and aggrecan gene expressions were not obviously up-regulated. In group OA P3-BMSCs, type Ⅰ collagen gene expression level lower than the control group in 3, 7, 14 day. In normal PO-BMSCs, GAG content showed 2.59-fold increase over that of the control group. GAG content of group OA PO-BMSCs and control group showed no significant differences.Others groups showed significant differences in comparison with the control group(P＜0.05). Alcian stain showed positive in all groups. The normal PO-BMSCs group showed the darkest blue-stained. Conclusion Rabbit normal PO chondrocytes and rabbit OA P0 chondrocytes significantly enhanced chondrogenic differentiation of rabbit BMSCs. The secreted morphogens of the rabbit normal P3 chondrocytes and rabbit OA P3 chondrocytes do not have a significant effect on chondrogenic differentiation of rabbit BMSCs.

A rare case of anaplastic lymphoma kinase(ALK)-negative anaplastic large cell lymphoma (ALCL)with generalized cutaneous involvement is reported in a 37-year-old man.Seven months prior to the presentation,he developed a goose egg-sized mass in his right thigh without obvious triggers,which gradually grew and no significant discomfort was felt.Diffuse and nonpitting edema gradually appeared in his right thigh and hip.Two months prior to the presentation,multiple dark red papules,nodules,and plaques emerged over the body surface with erosions and ulcers of varying size arising on some of the plaques.Laboratory examination revealed reduced albumin and significantly elevated lactate dehydrogenase in serum.B-mode sonography showed swelling and mutual fusion of superficial lymph nodes,and color Doppler flow imaging revealed markedly increased branch blood flow signals in lymph nodes.Computed tomography(CT)displayed generalized swelling of lymph nodes associated with soft-tissue edema in the right thigh and perineal region,as well as extensive enlargement of epigastric and mediastinal lymph nodes.Pathological examination of the skin lesion revealed a dense dermal infiltrate with mononuclear cells,some of which presented with cellular atypia and atypical nuclear division.Immunohistochemistry of the skin lesion showed that the mononuclear cells stained positive for CD3,CD8,CD30(80％ positive),CD4,CD45RO and granzyme B,but negative for CD56,ALK and T cell intracellular antigen-1(TIA-1).Pathology of lymph nodes indicated that the lymph node structure was completely destroyed with a diffuse growth of tumor cells,which were larger than common large cell lymphoma cells,and contained basophilic or bi-color abundant cytoplasm,deviating,horseshoe-,kidney-shaped,or lobulated cell nuclei,sparse nuclear chromatin and single or multiple small basophilic nucleoli.Angiogenesis,stromal fibrosis and infiltration of varying number of plasma cells and lymphocytes were seen in pathological lymphoid tissue.Immunohistochemistry of lymph nodes showed that the tumor cells stained positive for CD2,CD4,CD3,granzyme B,epithelial membrane antigen(EMA),Ki-67 and CD30,but negative for CD8,CD56,TIA-1 and ALK.The patient was diagnosed with ALK-negative primary systemic ALCL with extensive cutaneous involvement.

There are many physical factors affecting the development of cartilage tissue,the mechanical con-dition is the main important one that particularly act.The mechanical conditions used in engineering cartilage tissue,such as compressive and shear force,fluid flow,hydrostatic pressure and tissue deformation or with some of them combined,were reviewed.From the standpoint of bionics,the mechanical environments ap-plied on tissue engineering should work in three aspects:providing adequately mechanical stimuli to the cells seeded in 3-D scaffold;ensuring the efficient mass-transport of the nutrients and waste products in the cells:promoting the development of functionally extracellular matrix in 3-D scaffold.The mechanical environments currently used only represented the part of mechanical conditions of in vive articular cartilage will be reviewed.In our view that rolling depression load may achieve the fit mechanical environment for cultivation of functional cartilage constructs in vitro.

Objective To assess the quality of life, prevalence of depression and their influencing factors in patients with alopecia, to investigate, and to provide evidences for relevant clinical therapeutic strategies to improve patients′ quality of life. Methods A questionnaire survey was carried out in 237 patients with androgenetic alopecia or alopecia areata, and their quality of life and depression were measured using dermatology life quality index(DLQI)and center for epidemiologic studies depression scale (CES-D), respectively. Factors influencing the quality of life and depression were analyzed by analysis of variance and logistic regression analysis. Results Among 237 patients with alopecia, 218 questionnaires were eligible with the mean score of DLQI being 9.1 ± 5.4. Alopecia had a moderate effect on the quality of life in general, and 38.07%of the patients were severely affected. The mean score of CES-D was 14.8 ± 9.9, and 37.61%of the patients showed depressive tendency. The DLQI score was positively correlated with CES-D score(r=0.29, P<0.01). One-way analysis of variance(ANOVA)showed that the DLQI score was not affected by age, gender, education level or the number of visits. Multivariate logistic regression analysis revealed that the risk factors for depressive tendency in patients with alopecia were the number of visits (OR = 1.81, 95% CI: 1.21- 2.69) and DLQI score (OR = 1.08, 95% CI: 1.03- 1.13). Conclusion Alopecia not only affects the quality of life, but also mental states of patients.

BACKGROUND:Joint immobilization is one of the methods used to treat joint pain and joint injury in the department of orthopedics. Compared with other treatment methods, immobilization can reduce the pain of the damaged synovial joints and avoid the contact stress and friction between the joints. However, immobilization can cause some serious complications such as joint contracture, osteoporosis and cartilage degeneration.
OBJECTIVE:To observe the effects of joint immobilization on the repair of cartilage injury of knee joint in rats.
METHODS:Osteochondral ful-thickness defects (2.5 mm in diameter;2 mm in depth) were created in the left femoral condyle fossa with a corneal trephine. 36 animals were randomly assigned into immobilization group and control group (n=18 per group). In the control group, animal models were established, without any treatment. In the immobilization group, after model establishment, rats were immobilized by a designed and modified simplified miniature Ilizarov fixator.
RESULTS AND CONCLUSION:(1) Repair rate of cartilage defect:No significant difference in repair rate was detected between immobilization group and control group. (2) Histological staining:Regeneration tissue was mainly fiber cel s in both groups. At 8 weeks after surgery, Wakitani score and Mankin score were higher in the immobilization group than in the control group (P<0.05). (3) Cartilage metabolic marker detection:Compared with the control group, at 8 weeks, C-telopeptide of type II col agen levels in the urine were significantly higher in the immobilization group than in the control group (P<0.05). (4) Results indicated that persistent immobilization could result in cartilage degeneration, and it was detrimental for cartilage repair.

Crohn's disease (CD) is a chronic nonspecific intestinal inflammatory disease with unclear etiology and pathogenesis. At present, it is believed that CD can cause different degrees of metabolic abnormalities, and that the related metabolites also play an important role in the development and progression of the disease. In recent years, metabonomics is becoming more and more well-developed and has provided guidance for clinicians on rational nutritional support. Here is a review on the metabolic changes of multiple systems in CD patients, the application of metabonomics in CD diagnosis and treatment, and the research progress of clinical nutrition intervention.

Objective:To explore the risk factors for urinary tract infection (UTI) after a spinal cord injury (SCI).Methods:The medical records of 403 SCI patients were analyzed retrospectively. They were divided into UTI group and no-UTI group according to whether they had a UTI at admission. Gender, age, cause of injury, injury level of the spinal cord, voluntary anal contraction, time from injury to admission, American Spinal Injury Association (ASIA) grade, axillary temperature at admission, complications at admission (diabetes, hypertension, fracture of the pelvis, pressure sores or anemia), white blood cell count and urinary bacteria were compared between the two groups. Binary logistic regression was used to highlight the risk factors for a UTI after an SCI.Results:Of the 354 patients included in the final analysis, 62 (17.51%) had a UTI at admission. The regression showed that UTI after an SCI was closely related to an inability to voluntarily contract the anus, anemia, elevated white blood cell count and a high level of bacteria in the urine.Conclusions:Inability to contract the anus, fever, anemia and an elevated white blood cell count are independent indicators of a UTI after an SCI. A temperature ≥37.3°C is a simple indicator of a concentration of bacteria in the urine ≥1266/μL.