Objective To investigate the changes of interleukin(IL)-2 and interferon(IFN)-γin serum and synovial fluid of patients with joint injure with Kashin-Beck disease(KBD), and study the roles of IL-2 and IFN-γ in KBD joint injure. Methods In accordance with the "Diagnostic Criteria of Kashin-Beck disease"(GB16003-1995),48 cases of KBD patients and 26 healthy people(control group) from KBD endemic area in Long county Shaanxi province were enrolled in the study. KBD patient were 24 males and 24 females, respectively, aged 40 to 65 years (mean age 51 years). Forty-eight serum specimens and 28 synovial fluid specimens of patients(14 males and 14 females,respectively) were collected. Healthy control group were 13 males and 13 females, respectively. Twenty-six serum specimens of healthy controls were collected. Serum and synovial fluid IL-2 and IFN-γ levels were determined by enzyme-linked immunosorbent assay(ELISA). Results In healthy controls and KBD patients, the midian of serum IL-2 were 46.8 ng/L and 55.7 ng/L, respectively, and IFN-γ were 52.3 ng/L and 48.8 ng/L, respectively. The difference was not statistically significant between healthy controls and KBD patients(t = 0.62, 0.70, all P ＞ 0.05).In synovial fluid of KBD patient, the midian of IL-2 and IFN-γwere 48.3 ng/L and 44.1 ng/L, respectively. The difference was not statistically significant between serum and synovial fluid in KBD patients(t = 0.69, 1.72, all P ＞0.05). Conclusion Serum and synovial fluid IL-2 and IFN-γare not significantly increased in KBD patients with articular damage, indicating that IL-2 and IFN-γare not involved in KBD joint injury.

Objective To study the anti-tumor effects of griseofulvin on human prostate cancer PC-3 cells both in vitro and in vivo. Methods PC-3 cells were treated with griseofulvin at various concentrations for48 h,cell survival rate was then measured by MTT assay.The changes of morphology were observed by fluorescence microscope; Annexin V-FITC apoptosis detection kit was used to detect apoptosis of the cells ; The enzyme activity changes of caspase-3,8,9 were detected by spectrophotometry.For in vivo study,we first established the PC-3 tumor model by grafting PC-3 cells in athymic nude mice,and then injected griseofulvin into the tumors.12 days after injection,the mice were sacrificed,the tumors were removed,weighed and the ratios of tumor-suppression were then calculated.We had detected the expressions of Bcl-2,Bax,p53 and Survivin with immumohistochemistry as well. Results MTT results showed that griseofulvin could significantly inhibit the proliferation of PC-3 cells in vitro in a dose-dependent manner,and the IC50 of griseofulvin was 18.17 ±2.10 μg/ml.Typical morphological changes of PC-3 cells were observed by microscope.The rates of apoptosis of griseofulvin treated PC-3 cells greatly increased compared with that of the control cells (31.37 ± 2.93％ vs 2.89 ± 0.67％,P ＜ 0.01 ).The caspase-3,caspase-8 and caspase-9 activities in griseofulvin treated PC-3 cells were significantly higher than those in control cells (0.562 ±0.050 vs0.113±0.014,0.337±0.053 vs 0.120±0.017,0.293±0.038 vs0.109±0.018,P＜0.01).On the 23th day after tumor vaccination,the tumor volume was 961.17 ± 78.12 mm3 in griseofulvin treated group and was 433.6 ± 12.8 mm3 in control group (P ＜ 0.01 ).The tumor weight was 742.50 ± 78.63 mg in griseofulvin treated group and was 1387.33 ± 71.47 mg3 in control group ( P ＜ 0.01 ).Bcl-2,Bax,p53 and Survivin protein expressions were 16.10 ± 3.45％,39.50 ± 6.88％,48.20 ± 8.04％,16.50 ± 2.22％ in griseofulvin treated group,respectively; 41.30 ± 3.95％,13.70 ± 2.98％,17.60 ± 3.21％,52.11 ± 6.28％ in control group,respectively.And there were significant differences in both groups (P ＜ 0.01 ).The in vivo data showed that griseofulvin suppressed the tumor growth conspicuously through down-regulating the expression of Bcl-2,Survivin,and up-regulating the expression of Bax,p53. Conclusions Griseofulvin can inhibit the growth of PC-3 and induce apoptosis of PC-3 cells.Griseofulvin inhibits the in vivo tumorigenicity of PC-3 cells.

OBJECTIVETo study the related risk factors for surgical site infection following Pilon fracture surgery. METH ODS: The data of 561 patients with Pilon fractures treated with open reduction plate osteosynthesis at our institution's trauma centre were collected from January 2006 to December 2012. All the patients were divided into two groups: infection group and non-infection group. In the infection group, there were 23 males and 10 females, ranging in age from 21 to 69 years old, with an average of (45.50±4.40) years old. In the non-infection group, there were 296 males and 232 females, ranging in age from 16 to 76 years old, with an average of (43.50±7.19) years old. The possible risk factors such as age, gender, smoking, diabetes, alcohol abuse, open fractures, compartment syndrome and operative time were studied. The multivariate Logistic regression model was used to analyze the risk, factors.

RESULTSThe infection rate of surgical site after Pilon fracture surgery was 5.88%. There were significant statistical differences between infection group and non-infection group in operative time, open fractures and compartment syndrome. However, multivariate Logistic regression analysis revealed that only operative time was significantly associated with surgical site infection (P=0.005, OR=44.92).

CONCLUSIONOperation time is an independent predictor for post-operative surgical site infection of Pilon fracture treated with open reduction plate osteosynthesis. Though open fracture and compartment syndrome could increase the surgical site infection rate, they could not not be considered as independent predictors.

Adult ; Compartment Syndromes ; complications ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Operative Time ; Risk Factors ; Surgical Wound Infection ; etiology ; Tibial Fractures ; surgery

Objective To investigate the effect of different Chinese herbs on cell proliferation and cartilage oligomeric matrix protein(COMP)expression in chondrocyte culture.Methods Chondrocytes isolated from rabbit knee cartilage were cultured for 3 generations with the density of 2?10~4/cm~2 and were verified by collagenⅡimmunohistochemical staining.Rabbit sera containing herbs were obtained after animals orally ad- ministrated herbs at the dosage equivalent to human.At 5% and 10% serum density,cells were cultured in the medium that contained liver-softening herbal compound sera.Subgroups setting at 1,3 and 5 hours after herb intervention were observed.Rabbit and bovine sera were control groups.Seven days after intervention,chon- drocytes proliferation was observed using the MTT assay kit.For the study of COMP expression,chondrocytes were isolated from human knee cartilage supematant.Superuatant COMP level was tested by enzyme-linked immunoabsorbent assays(ELISA)after directly adding compound and extract from liver-softening herbs to the culture at the final concentration of 10 mg/ml for 3 days.Results Liver-softening herbal compound group had significant effect on cell proliferation compared to control,of which,3-hour subgroup was more significant than 1-and 5-hour subgroups(P

Objective To investigate the feasibility of urethral reconstruction with colonic mucosa graft in the treatment of complex long urethral stricture.Methods The clinical data of three cases with complex long urethral stricture were reported and analyzed.Patient ages were 71,64 and 48 yrs and the course of disease was three months,six months and six yrs,respectively.The length of urethral stricture was 13,18 and 12 cm.Removing the narrow urethral segment and intercepting the length from 12 to 18 cm sigmoid colon and stripping colonic mucosa were performed.Urethral reconstruction was done with a free graft of colonic mucosa.Follow-up included urethrography,uroflowmetry,and urethroscopy.Results The urethral reconstructions were completed successfully.The urinary peak flows of the patients were 16.7 ml/s,19.6 ml/s and 26.4 ml/s at six weeks post operation.Urethrography revealed the graft urethral lumens were bulky three months after the operation.In urethroscopy,the colonic mucosa was found to be of good color and the anastomotic site healed well.Patients were followed-up 28,16,and three months,respectively,and were all voiding well.Conclusions Colonic mucosa graft urethroplasty is a feasible procedure for the treatment of complex long urethral stricture.

AIM To investigate the effects of melatonin on rats colitis induced by acetic acid and its mechanism. METHODS The rats colitis model was produced by acetic acid enemas. The experimental animals was divided into 6 groups: normal group,model group,5 ASA group(100 mg?kg -1 ),MT group(2 5,5 0,10 0 mg?kg -1 ), and treated intracolonic with saline,5 ASA and MT correspondingly (1 time per day, from the 24 h after the establishment of colitis model to the end of the experiment). At the end of the experiment, the mucosal damage index(CMDI),the level of myeloperoxidase(MPO), the score of histology(HS) in rats colon and the occult blood test(OB) in feces were observed and measured, meanwhile, the content of malondiadehyde(MDA), glutathione peroxidase(GSH Px),catalase (CAT), superoxide dismutase(SOD), nitric oxide(NO) in rats colon were detected. RESULTS After being treated with acetic acid, the exetent of OB, CMDI, HS and the level of MPO in model group were more higher than that in normal group, MT(2 5,5 0,10 0 mg?kg -1 ) ameliorated the exetent of CMDI and OB and reduced the level of MPO, MT(10 0 mg?kg -1 ) alleviated the HS of rats in model group. Meanwhile in model group, the content of MDA,NO in colon elevated, but the content of GSH Px,SOD and CAT in colon reduced remarkably. MT intracolon depressed the content of MDA,NO in colon, promote the contents of GSH Px?SOD and CAT in colon. CONCLUSION Treatment with MT shows beneficial effects on the mucosal damage of rats colitis induced by acetic acid.

ED are correlated with the severity of CAD in a degree.

Objective:To study the cytocompatibility of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation.Methods:Components of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation in three different voltages of 300 V,350 V and 400 V,Zr-Cu-Al-Ag alloy as cast condition and TI6Al4V alloy were made for the test.The water extracted from the components were obtained according to national standard.The L929 cells were cultivated in vitro in the extracts of these components separately.The L929 cells,cultured in Dulbecco's modified Eagle medium supplemented with 10 ％ fetal calf serum,served as the negative control group.And cells,cultured in Dulbecco's modified Eagle medium supplemented with 10 ％ fetal calf serum and 64 g/L phenol,served as the positive control group.The cytocompatibility of these components were evaluated by MTT colorimetric.Results:The cytotoxicity of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation is 0 grade.Microscopy showed that the morphology of L929 cells,cultured in the extracts of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation were normal.There were no significant differences between micro-arc oxidationt and negative control groups.The cell multiplication curves of micro-arc oxidation and negative control groups were nearly overlapping and in the linearity increasing trend.The OD in micro-arc oxidation groups had no significant differences with negative control group (P＞0.05),but were higher than that of Zr-Cu-Al-Ag alloy as cast condition,TI6Al4V alloy and positive control groups (P＜0.05).Conclusions:The cytocompatibility of Zr-Cu-Al-Ag alloy has been improved by micro-arc oxidation technique.

Objective To investigate the effect of selenium on proliferation and apoptosis of chondrocytes of articular cartilage cultured in vitro in Kaschin-Beck disease(KBD) patients and normal person, to explore the role of selenium in control of KBD, and to provide evidence for selenium's effect on the growth of normal cartilage cells. Methods The articular cartilage samples of grade Ⅱ and Ⅲ KBD patients were selected according to the national "Clinical Diagnosis of KBD" (GB 16003-1995). Chondrocytes of 5 KBD and 5 non-endemic normal accidentswere separated and cultured in vitro. KBD group and control group were given different doses of selenium (0,0.0125,0.0250,0.0500,0.1000,0.2500,0.5000,1.0000 mg/L, respectively). Methyl thiazolyl tetrazolium (MTT),flow cytometric analysis, and immunocytochemical staining were used to observe the effect of selenium on cell growth and apoptosis in KBD and normal persons. Results MTT results showed that the cell proliferation rate in each dosage group of the control group at the 6th day(0.086 ± 0.025,0.077 ± 0.012,0.073 ± 0.027,0.071 ± 0.017,0.058 ± 0.028,0.052 ± 0.028 and 0.046 ± 0.037) was significantly lower than that of 0 mg/L group(0.138 ± 0.026,all P ＜ 0.05);the average cell proliferation rate was negative( - 0.001 ± 0.001, - 0.003 ± 0.000, - 0.003 ± 0.001and - 0.004 ± 0.001 ) in 0.1000 - 1.0000 mg/L dose group, which was significantly lower than that of the 0 mg/L group(0.025 ± 0.003, all P ＜ 0.05);compared with 0 mg/L group(0. 115 ± 0.011), the KBD 0.2500 mg/L dose group promoted cell proliferation(0.128 ± 0.037, P ＜ 0.05), the KBD 1.0000 mg/L dose group inhibited cell growth (0.071 ± 0.019, P ＜ 0.05). The apoptotic rate of 0.0500 - 1.0000 mg/L dose control group [ (18.88 ± 0.02)%,(17.58 ± 0.01)%, (17.09 ± 0.04)%, (56.00 ± 0.02)%, (57.85 ± 0.03)% ] were higher than that of the 0 mg/L group[(13.51 ± 0.01)%, all P ＜ 0.05];compared with 0 mg/L group[(25.84 ± 0.02)%], the apoptotic rate in KBD 0.0250 - 0.2500 mg/L dose group [ ( 13.69 ± 0.02) %, ( 15.96 ± 0.03 ) %, ( 16.68 ± 0.03 ) %, ( 16.67 ± 0.02) % ]were lower, and the apoptotic rate in 0.5000, 1.0000 mg/L dose group [ (59.58 ± 0.03)%, (73.48 ± 0.04)% ] were significantly higher(all P ＜ 0.05). The Fas expression in KBD 0.0500 - 0.2500 mg/L dose groups[ (41.2 ± 1.5)%,(40.3 ± 2.0)%, (50.2 ± 2.5)%] were lower than those of the same dose control group with selenium intervention [(52.4 ± 1.0)%, (67.2 ± 4.0)%, (75.1 ± 5.0)%, all P ＜ 0.05], the caspase-3 expression in KBD 0.0500,0.1000 mg/L dose groups[ (40.8 ± 1.1 )%, (45.1 ± 2.1 )%] were lower than those of the same dose control group with selenium intervention[ (68.0 ± 3.0)%, (70.6 ± 3.5)%, all P ＜ 0.05 ]. Conclusions Appropriate dose of selenium supplementation (0.1000 - 0.2500 mg/L) could promote the growth of KBD chondrocyte, decrease cell apoptosis,but have a damage when the dose of selenium ＞ 0.5000 mg/L;doses of selenium that could promote the growth of KBD chondrocyte does not mean to promote the growth of normal cartilage cells in vivo.

Objective To investigate whether rat adipose-derived stem cells (rASCs) could resist human xenoantibody-dependent complement-mediated lysis and to explore its possible mechanisms.Method SD rat ASCs were isolated,rASCs at passage 2 to 8 were used for the following studies and rat lymphocytes were harvested as control cells.α-Gal expression was detected by flow cytometry.After incubation of rASCs with 20％ normal human serum (NHS) or heat inactivated normal human serum (HINHS),flow cytometry was used to detect cytotoxicity,IgG or IgM binding,and C3c,C4c and C5b-9 deposition.Result We successfully established the method to isolate and culture rASCs.The morphology of rASCs remained unchanged after passages.rASCs were positive for tell surface markers of CD44 and CD90,while negative for CD45 and MHC-Ⅱ.As compared with rLCs,rASCs significantly resisted human natural antibody and complement-mediated lysis when incubated with 20％ NHS in vitro (20.42％ ± 2.80％ vs 51.84％ ± 6.70％,P ＜ 0.01).Mechanistically,rASCs expressed lower level of α-Gal (13.97 ± 0.33 vs.24.47 ± 3.03,P＜0.05),which was correlated with decreased binding of human xenoreactive IgG and IgM (IgM:9.4 ± 2.0 vs.107.2± 4.8,P＜0.01; IgG:5.73 ± 1.0 vs.27.49 ± 3.9,P＜0.01) and reduced deposition of complements C3c,C4c and C5b-9 (C3c:294.6 ± 38.02 vs.1924 ± 509.4,P＜0.05; C4c:35.23 ± 3.1vs.177.3 ± 37.17,P＜0.05; C5b-9:5.63 ± 1.74 vs.37.05 ± 7.4,P＜0.01).Conclusion These data demonstrated that the resistance of rASCs to human xenoantibody and complement-mediated lysis is associated with low expression of xenoantigen a-Gal and inhibition of MAC (membrane attack complex) formation.