BACKGROUND:Apoptosis,which result in various reasons,plays an important role in intervertebral disc degeneration. One of important reasons for apoptosis is oxidative stress. OBJECTIVE:To investigate effects and mechanism of hydrogen-peroxide(H2O2) on nucleus pulposus cell injury induced by oxidative stress at intracellular signal transduction level in rats. DESIGN,TIME AND SETTING:A single sample observation was performed at the institute of Traumatology and Orthopaedics of Shandong Province from March to October in 2008. MATERIALS:The specific p38 mitogen-activated protein kinase(p38 MAPK) inhibitor SB203580,the specific JNK inhibitor SP600125 were purchased from Beyotime Institute of Biotechnology;Two 24-hour-old SD rats of SPF degree were purchased from Qingdao Institute for Drug Control. METHODS:The primary cultured nucleus pulposus cells were divided into four groups:the H2O2 group,stimulated by H2O2 with concentrations of 0,50,100,200,400,and 800 ?mol/L;Control group;SB203580+H2O2 group:cells were preincubated with SB203580,and then were treated by stimulated by H2O2;SP600125+H2O2 group,cells were preincubated with SP600125,followed by stimulated by H2O2. MAIN OUTCOME MEASURES:The expression and cellular localization of P-p38MAPK and P-JNK was detected by immunohistochemistry,and the expression of total and phosphorylated SAPK/JNK,p38MAPK were measured by Western blotting. RESULTS:H2O2 could activate the activity of P38 and JNK. The expression of P38MAPK was significantly inhibited with the pretreatment of SB203580;however,the SP600125 could inhibit the expression of JNK. Immunofluorescence analysis demonstrated that P-p38MAPK and P-JNK were expressed and distributed mainly in cytoplasmic and nuclear exception of the control group. CONCLUSIONS:Apoptosis of rat nucleus pulposus cells are induced by oxidative stress via p38MAPK and JNK signal pathway.

Objective To prepare gastric floating sustained-release tablets of Triperygium wilfordii and study its behavior of floating and release characteristics. Methods The sustained-release tablets were prepared by direct powder compared to tablets technique. The floating and release of total diterpene-lactones were used as indicators to evaluate and optimize the formulation. Then the formulation was optimized by influential factors and orthogonal design test. Results The gastric floating sustained-release tablets which was taken orally twice one day, were prepared with HPMCK4M as matrix, cetyl alcohol as floating assistant, sodium bicarbonate as gas-producer, PVP as poremaking. The tablets released 30% in 2 h, 60% in 6 h, exceeding 90% in 12 h, the release behavior of the tablets was fitted to Higuchi equation,and it was properly characterized by the drug diffusion and bulk erosion mechanism. Conclusion Gastric floating sustained-release tablets of T. wilfordii prepared has good behavior of floating and release characteristics.

Objective To investigate the drug in vitro release behavior of gastric floating sustained-release preparation of Tripterygium wilfordii and establish their quality evaluating methods.Methods HPLC was Employed to gain the fingerprints of releasing medium of preparations.Triptolide was selected as marker to calculate the linear equation concerning peak area then the relative drug release and f2 simila-rity factor value were acquired.Results Various components in gastric floating sustained-release tablets and pellets of T.wilfordii could not release synchronously while sustained-releasing but all of them in the gastric floating sustained-release capsules of T.wilfordii prepared by using multiparticulate site-controlled release technology could release synchronously while sustained-releasing.Conclusion The quality and in vitro release behavior of gastric floating sustained-release preparation of T.wilfordii could be evaluated more scientifically and comprehensively by using HPLC fingerprints method.

Objective To investigates the diagnostic value of CT of the gallbladder and accuracy.Methods Clinical data of 26 patients with pathologically confirmed gallhladder were retrospectively analyzed.Results 26 patients were diagnosed with CT.CT manifestations of primary gallbladder carcinoma:papillary intraluminal mass type,limited or diffuse gallbladder wall thickening,large cystic mass in full or swallowed type.Combined with the direct signs of pathology gallbladder cancer was divided into three types:infiltrative,nodular,tumor type;Enhance a performance of CT scan of gallbladder:artery of high density,such as density of the portal vein,followed by arterial and portal vein of both high-density,higher than normal latency of the gallbladder wall and liver,regression not significant,especially gallbladder junction local thickening of the gallbladder wall continued enhanced features.Conclusion The performance of primary gallbladder was complex,and CT examination was a key means of establishing diagnosis.

Objective To evaluate the feasibility of retroperitoneal laparoscopy for patients with upper ureteral calculi. Methods A total of 35 patients with upper ureteral calculi were treated with retroperitoneal laparoscopy. Results The operation was completed in all but one patient, who was converted to open surgery because the calculi moved into the renal pelvis. The operation time ranged from 70 to 135 minutes (mean, 110 minutes). Intraoperative blood loss was 20 to 55 ml (mean, 36 ml). No patient had over-2-day urine leakage. Retroperitoneal drainage tube was removed 2 to 3 days after the operation. The postoperative hospital stay ranged from 6 to 10 days (mean, 7.8 days). One month after the operation, when double-J catheter was removed, ultrasonography showed that 9 cases who had severe hydronephrosis before operation was relieved. In the 26 patients with mild or moderate hydronephrosis, the symptoms disappeared in 17, and were relieved in the other 9. Seven patients with severe hydronephrosis and 11 patients with mild or moderate hydronephrosis achieved a 2-to 6-month follow-up, none of them developed recurrent calculi during the period. The severity of hydronephrosis in these patients was same to that determined one month postoperation. Conclusions Retroperitoneal laparoscopy is feasible for patients with upper ureteral calculi. The method can be used as an alternative to open surgery of microinvasive operation.

Objective To study the proliferation and apoptosis of related proteins in pathological liver tissues of alcohol-induced mice,and establish a model and time-evolution rule of liver cell apoptosis,which can be used to guide the clinical treatment of acute alcoholic liver injury.Methods A total of 30 male KM mice were fed in a clean grade animal room at the Capital Medical University and then randomly (random number) separated into two groups.The 10 mice in the normal group were fed without ethanol,while the other 20 mice in the experimental group were given a one-time grant of 50％ ethanol (12 mL/kg) by gavage.The mice in the experimental group were killed at two time points,6 h for 10 mice and 12 h for the other 10,after the intragastric administration.Hematoxylin and eosin (HE) staining was used to observe the morphological changes of liver in mice.The concentrations of T-ERK,p-ERK,PKC,p-PKC and caspase-3 were determined by the Western-blot method.The data were analyzed by Analysis of variance (ANOVA) method using statistical software SPSS 11.5 and criterion P ＜ 0.05 is chosen to determine differences that are statistically significant.Results By observing the behavioral changes and morphological indexes of mice,we confirmed the success of the model for acute alcoholic liver injury.During the process of re-building the model,no mice died.The mice in the experimental group appeared in drunken states,such as sleepiness and slowness of movement.Compared to the normal group,the experimental subgroup at the 6 h point showed no difference statistical significant; while the experimental subgroup at the 12 h point showed obvious histological changes in tissues,including the disorder of hepatic lobule structure and fatty vacuolization of hepatocytes.At the same time,in the experimental subgroup at the 12 h point,both P-ERK and P-PKC significantly decreased [(2.41 ±0.38),(0.97 ±0.25),F=4.82,P＜0.05; (0.16 ±0.00),(0.08 ± 0.01),F =29.63,P ＜ 0.05],but caspase-3 significantly increased [(0.30 ± 0.02),(0.11 ± 0.01),F =34.38,P ＜ 0.05].Conclusions In mice after intragastric administration of large doses of alcohol,the hepatic cell apoptosis appeared mainly after 6 h but before 12 h,therefore 6 ～ 12 h might be the time window to inhibit the cell apoptosis of mice' s acute liver injury from alcohol induction.

ObJective To locate the cysteine-rich domains(CRD) of murine 4-1BB binding to its natural ligand. Methods A serial soluble extracellular CRDs of routine 4-1BB and 4-1BBL fusion proteins was constructed and prepared. The binding of purified 4-1BB-Igs to 4-1BBL and 4-1BB monoclonal antibody were tested using ELISA assay and Western blot analysis. Blocking experiment with 4-1BBL and 4-1BB mon-oclonal antibody was performed by ELISA assay. Results All truncated overlapped proteins containing ex-tracellular CRD Ⅱ of murine 4-1BB were able to bind to 4-1BBL by ELISA assay, excepting the CRD Ⅰ do-main alone. A 4-1BB monoclonal antibody proved to block the interaction of 4-1BB and 4-1BBI, was also able to bind to CRD Ⅱ. Conclusion Murine 4-1BBL whose specificity was mapped to CRD Ⅱ of 4-1BB ex-tracellular region with a possible conformational structure.

OBJECTIVE:To investigate the recent use of immunosuppressive agents for renal transplantation patients during our hospital.METHODS:Case history of124renal transplantation patients in our hospital in2004was reviewed ret-rospectively,and the categories and the associated costs of immunosuppressive agents were statistically analyzed,which were compared with that of2000.RESULTS:In2004,patients’mean hospitalization cost was74462.77yuan and mean immuno-suppressive agents cost was25809.93yuan,increased by75.12%and175.35%,respectively as compared to2000.Of the total drug costs,the immunosuppressive agents accounted for61.37%in2004and46.4%in2000.Dominated the first3places in the list of DDDs of immunosuppressive agents in2004were Methylprednisolone,ciclosporin A and dehydrocortisone,respec-tively.CONCLUSIONS:As compared with2000,the application categories and costs of neotype immunosuppressive agents for renal transplantation patients in our hospital increased in2004,thus to find a more effective immunosuppressive remedy that of less adverse drug reaction and lower cost is of great significance.

BACKGROUND:Scholars have studied the three-dimensional finite element models of tibial plateau fractures with different fixation methods, but the research on three-dimensional finite element model of tibial plateau fracture fixation was not much. OBJECTIVE:To analyze the biomechanical characteristics of external fixator for tibial plateau fracture by using the three-dimensional finite element method. METHODS:The tibial plateau fracture models treated with locking plate fixation and external fixator were established. Three-dimensional finite element analysis was used to analyze the stress distribution and displacement of two kinds of models. RESULTS AND CONCLUSION:(1) The average displacement value and the maximum displacement value of tibial plateau fracture external fixator model group were smal er than that of tibial plateau fracture locking plate fixation model group, but no significant difference was found (P>0.05). (2) X axial displacement value and Y axial displacement value were smal er in the tibial plateau fracture external fixator model group than in the tibial plateau fracture locking plate fixation model group, but no significant difference was found (P>0.05). (3) The stress of tibial plateau fracture locking plate fixation model focused on the junction of the screw and bone tissue as wel as the junction of plates and screws. The maximum stress value of tibial plateau fracture locking plate fixation model (173 MPa) was greater than that of tibial plateau fracture external fixator model (86 MPa). The stress of tibial plateau fracture external fixator model was scattered. Obvious stress concentration did not occur. The stress at the junction of the clamp and the nail and the junction of the connecting rod and screw was higher than that at other regions, and was a weak link in the clinical mechanics. (4) Results showed that external fixation for tibial plateau fractures had better stability and fixing strength as compared with the internal fixation.

Objective:To investigate the values of immunophenotype and the Collagen type1 alpha1/Proto-oncogene Proteins c-sis (COL1A1/PDGFB) fusion gene in the diagnosis of dermatofibrosarcoma protuberans (DFSP). Methods:IHC markers and the COL1A1/PDGFB fusion gene were detected by IHC staining and interphase fluorescence in situ hybridization (FISH) in 73 cases previously diagnosed as DFSP. A total of 85 and 10 non-DFSP cases were also included as controls for IHC staining and FISH, respectively. Results:In the 73 DFSP cases, the positive detection rates for immunohistochemical marker vimentin, CD34, CD99, S100, desmin and SMA were 100%, 91.78%, 61.64%, 0, 0, and 6.85%, correspondingly. Protein expression levels in these cases varied from the control group, and CD34 ex-pression was significantly different among the differential diagnoses. The positive detection rate for the COL1A1/PDGFB fusion gene was 86.96%(60/69), whereas the gene expression in the control group was negative. Conclusion:The COL1A1/PDGFB fusion gene is a highly specific and sensitive marker in the diagnosis of DFSP. CD34 is a suitable marker for DFSP.