AIM To determine the relationship between the elevation of endogenous inhibitor of nitric oxide synthase (NOS)N G,N G-asymmetric dimethylarginine (ADMA) and metabolic control in diabetic rats. METHODS Diabetes was induced in Sprague-Dawley rats by a single intraperitoneal injection of streptozotocin. At 72 h after injection, half of diabetic rats received insulin treatment for 8 weeks (20 U?kg -1?d -1,ih, bid). Serum levels of ADMA were measured by high-performance liquid chromatography. Thoracic aortic rings from non-diabetic age-matched control, untreated diabetic, and insulin-treated diabetic rats were tethered in isolated organ baths,contracted with 1 ?mol?L -1 phenylephrine, and challenged with either the endothelium-dependent vasodilator acetylcholine or the endothelium-independent vasodilator sodium nitroprusside. Serum concentrations of glucose, glycosylated serum protein, and malondialdehyde, derived from lipid peroxidation were also examined to estimate metabolic control.RESULTS Serum levels of ADMA significantly elevated in untreated diabetic rats compared with control rats. This elevation of ADMA was accompanied by impairment of relaxation response to acetylcholine but not sodium nitroprusside in aortic rings. Chronic insulin treatment not only prevented the elevation of serum ADMA, but also improved the impaired endothelium-dependent relaxation in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialdehyde were significantly increased in parallel with the elevation of ADMA in untreated diabetic rats compared with control rats. These parameters were normalized after diabetic rats received insulin treatment. CONCLUSION These results provide the first evidence that the elevation of endogenous inhibitor of NOS in streptozotocin-induced diabetic rats is close related to metabolic control of the disease.

Objective:To explore the regulatory role of exosomes in calcification of rat vascular smooth muscle cells (VSMC) induced by high phosphorus.Methods:VSMC (A7r5 cells) were cultured in vitro and randomly divided into three groups: normal phosphorus group (0.9 mmol/L), high phosphorus group (2.6 mmol/L) and high phosphorus exosomes induction group (i.e. the exosomes extracted from high phosphorus group were added to VSMC in normal culture). Until the 7th day of culture, the culture medium of normal phosphorus group and high phosphorus group obtained during the change of cell culture medium was collected, and the precipitate was obtained by ultracentrifugation and suspended by phosphate buffered saline. The protein content of the precipitate was determined by BCA protein quantitative method. The precipitates were identified. The structure and size of exosomes were observed by transmission electron microscope. The exosomes marker proteins tumor susceptibility gene 101 protein (TSG101) and CD9 were detected by Western blotting. The miRNA in exosomes was extracted, and the expression of related miRNA (miR-30b, miR-204, miR-211) were observed by real-time quantitative PCR. After 7 days of cultivation, the exosomes uptake process of VSMC in high phosphorus exosomes induction group was observed. The calcium deposition was detected by Alizarin stain, and the calcium content was detected by O-cresol complex copper. The content of alkaline phosphatase was detected by colorimetry. The protein expression of Runx2 was quantified by Western blotting. Results:(1) The precipitate obtained by ultracentrifugation of the cell culture fluid was identified as exosomes by electron microscopy morphology. Western blotting confirmed that the expression of the exosomal marker proteins TSG101 and CD9 were positive. (2) The exosomes were rich in miRNAs. The expression of miR-30b, miR-204, miR-211, which negatively regulated the transcription of Runx2, was significantly down-regulated in the high-phosphorus group compared with the normal group ( P<0.05). (3) After culturing rat VSMC with high phosphorus for 7 days, calcium salt deposition was obvious. Compared with the normal phosphorus group, calcium content and alkaline phosphatase activity were significantly increased (both P<0.05), and Runx2 expression was also significantly increased ( P<0.05). (4) Added the obtained high-phosphorus exosomes to the normal cultured VSMC, the exosomes could be taken up by VSMC and successfully induced VSMC calcification. The levels of cell calcification indicators and Runx2 expression were significantly increased. Conclusions:High phosphorus induces calcification of VSMC and promotes the increase of Runx2 expression. The mechanism may be realized by releasing exosomes from VSMC to transmit cell signals.

Abstract:The present case report retrospectively analyzed 36 cases of injury to posterior pelvic dng who received treatment in the Department of Orthopedics,Yan'an Hospital between January 2004 and April 2007. According to the Denis classification,there were 20 cases of type I injury to posterior pelvic ring,12 of type Ⅰ,and 4 of Ⅲtype Ⅲ; according to Tile classification,there were 16 cases of B3,10 of C1,6 of C2,and 4 of C3. Pedicle screw fixation system was employed. Two curvy incisions were made on the bilateral posterior superior lilac spine and a hole was chiseled on each side to fit the pedicle screw. Then,a universal pedicle screw was driven between the inner and outer lilac plates until the screw were sunk. A rod was finally traversed through erector spinae to connect two pedicle screws. Thirty cases were available at follow-up with a mean of 11 months (range:6-11 months). None of 30 cases presented with nerve or vessel injury and breakage,loosening of screws,or redislocation of sacroiliac joints. According to functional rating scale,excellent results were found in 15 cases,good in 13,and fair in 2 cases. These findings suggest that treatment for injury to posterior pelvic ring with pedicle screw fixation system is simple and reliable with microtrauma and little bleeding; in addition,it affords good recovery and can be employed in primary hospital without monitoring of X-ray,so it will deserve popular practice.

AIM This study was designed to investigate the effect of aminoguanidine, an inhibitor of the glycosylated proteins formation, on the impairment of endothelium dependent relaxation induced by advanced glycation end products (AGE) in isolated rat thoracic aorta and its possible mechanisms. METHODS Exogenous glycosylated bovine serum albumin (AGE BSA) was prepared. Aortic rings were exposed to AGE BSA for 60 min to induce the impairment of endothelium dependent vasodilatation. In the drug treated groups, aortic rings were incubated with drug for 15 min and then exposed to AGE BSA for another 60 min in the presence of the drug. Vasodilator responses to acetylcholine (ACh) or sodium nitroprusside (SNP) of aortic rings were measured by isometric tension recording after drug treatment. RESULTS AGE BSA significantly inhibited the endothelium dependent relaxation in response to ACh, but did not affect endothelium independent relaxation in response to SNP. Pre incubation of aortic rings with aminoguanidine(50～500 ?mol?L -1 ) for 15 min and co incubation of aortic rings with AGE BSA for another 60 min markedly attenuated the inhibition of endothelium dependenet relaxation induced by AGE BSA in a dose dependent manner. Superoxide diamutase (SOD, 2?10 5 U?L -1 ), a scavenger of superoxide anions, also prevented the inhibition of endothelium dependent relaxation, which is similar to the effect of 500 ?mol?L -1 aminoguanidine. Furthermore, aminoguanidine (500 ?mol?L -1 ) also reversed impairment of endothelium dependent relaxation of rat aortic ring induced by endogenous oxygen free radicals generated by diethyldithiocarbamate (DETC, 10 ?mol?L -1 ) via inhibiting intracellular SOD. CONCLUSION Aminoguanidine can protect rat aortic endothelium against damage due to AGE BSA, and the beneficial effect of aminoguanidine may relate to its antioxidant properties.

BACKGROUND: Previous studies have confirmed that rabbit bone marrow mesenchymal stem cells (BMSCs) can differentiate into osteoblasts under osteogenic induction in vitro, stably express the specific phenotype of osteoblasts and have osteogenic ability. Calf cortical bone scaffold with partial cancellous bone has good biocompatibility and degradability, which can be used as a carrier material of bone marrow mesenchymal stem cells. OBJECTIVE: To combine rabbit BMSCs with calf bone composite according to the basic principles of bone tissue engineering and to observe the osteogenesis in the New Zealand white rabbits after implantation of BMSCs/calf bone composite into the ilium, thereby providing a direct evidence for preliminary clinical application of tissue-engineered bone products.METHODS: BMSCs/calf cortical bone scaffold with partial cancellous bone (tissue-engineered bone group), simple calf heterogeneous bone (heterogeneous bone group) or autologous iliac bone (autologous iliac bone group) was randomly implanted into the rabbit ilium. The changes of implant surface and tissue reactions around the implant were observed.X-ray examination was performed to observe osteogenic changes at 4, 8, 12, 24 weeks after implantation. Immunohistochemistry staining was used to observe the expression of bone morphogenetic protein 2.RESULTS AND CONCLUSION: After heterogeneous bone implantation, the wound healed well, and there were no systemic or local inflammation and toxicity reactions in all groups. The X-ray results showed that at postoperative 24 weeks, the implant was basically fused with the host bone in the tissue-engineered bone group, but the fusion was unsatisfactory in the heterogeneous bone group. The process of ossifications from cartilages was observed in all groups by hematoxylin-eosin staining, and bone morphogenetic protein 2 was positive for immunohistochemical staining. Findings from in vivo experiments indicate that rabbit BMSCs seeded onto the calf cortical bone scaffold with partial cancellous bone could construct tissue-engineered bone by osteoinductation in vitro in the rabbits.

Objective To detect the selective inhibitory effects of irradiation plus adenovirusmediated horseradish peroxidase ( HRP)/indole-3-acetic acid (IAA) suicide gene system using tumorspecific and radio-inducible chimeric promoter on human hepatocellular carcinoma subcutaneously xenografted in nude mouse.MethodsRecombinant replicated-deficient adenovirus vector containing HRP gene and chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 radioinducible CArG elements was constructed.A human subcutaneous transplanting hepatocellular carcinoma (MHCC97 cell line) model was treated with -γ-ray irradiation plus intra-tumor injections of adenoviral vector and intra-peritoneal injections of prodrug IAA.The change of tumor volume and tumor growth inhibiting rate,the survival time of nude mice,as well as histopathology of xenograft tumor and normal tissues were evaluated.Results Thirty one days after the treatment,the relative tumor volumes in the negative,adenovirus therapy,irradiation,and combination groups were 49.23 ± 4.55,27.71 :± 7.74,28.53 + 10.48 and 11.58 ± 3.23,respectively.There was a significantly statistical difference among them (F = 16.288,P ＜0.01 ).The inhibition effect in the combination group was strongest as compared with that in other groups,and its inhibition ratio was 76.5%.The survival period extended to 43 d in the combination group,which showed a significantly difference with that in the control group(x2 = 18.307 ,P ＜0.01 ).The area of tumors necrosis in the combination group was larger than that in the other groups,and the normal tissues showed no treatment-related toxic effect in all groups.However,multiple hepatocellular carcinoma metastases were observed in the liver in the control group,there were a few metastases in the monotherapy groups and no metastasis in the combination group.Conclusions Adenovirus-mediated suicide gene therapy plus radiotherapy dramatically could inhibit tumor growth and prolong median survival time.It might provide a promising therapeutic modality for hepatocellular carcinoma therapy.

Clinical data and manifestations on muhi-slice spiral CT (MSCT) of 11 patients with disseminated peritoneal adenomucinosis (DPAM) were retrospectively reviewed.The CT manifestations were also compared with surgical and histopathological findings.MSCT findings showed a large amount of gel-like ascites in 9 cases and local cystic masses in 2 cases.Among 9 cases with a large amount ascites,abdominal multiple cystic masses were shown in 5 case,and infiltration of the greater omentum and mesentery in 5 cases.Hepatic scalloping was found in 6 cases ; parenchymal invasion of the liver or spleen were showed in 5 cases; calcification of the cystic masses in 5 cases.Ovary mueinous cystadenoma was presented in 3 female patients.Enlarged lymph nodes and omental cake were not found in all cases.The results indicate that the characteristic MSCT manifestations of DPAM include diffuse gel-like ascites,multiple cystic masses with or without calcification,hepatic scalloping and parenchyma invasion.

The standard treatment for colorectal cancer still have not been unified,especially the treatment for patients with locally advanced rectal cancer (T3 or T4 stage) is controversial,and multidisciplinary comprehensive treatment is recommended by most of the scholars.Precise preoperative diagnosis and staging for locally advanced rectal cancer is very important.Currently,transrectal ultrasound (TRUS),multi-slice spiral computed tomography,magnetic resonance imaging (MRI) are playing important roles in the preoperative diagnosis and staging,while controversies on the efficacies of different imaging diagnosis on the evaluation of rectal cancer exist between domestic and overseas scholars.From October 2010 to October 2012,the imaging data of 30 patients with locally advanced rectal cancer who were admitted to the Tumor Hospital of Yunnan Province were retrospectively analyzed.The results showed that TRUS,computed tumography and MRI had guidance value in preoperative tumor staging and evaluation of neoadjuvant therapy for patients with locally advanced rectal cancer,which provide a more reliable basis for the selection of appropriate treatment programs.

ObjecfiveTo investigate clinical features and imaging manifestation in patients with posterior reversible eneephalopathy syndrome (PRES) to improve its recognition.MethodsSix patients with PRES were enrolled,four women with history of end-stage renal disease,kidney transplantation,eclampsia,or systemic lupus erythematosus (SLE) and two men with history of chemotherapy or hypertension.All of them underwent multi-serial MR imaging (T1 WI,T2 WI,FLAIR) and post-contrast T1 WI.Three cases also underwent CT scan and gadolinium enhancement. ResultsAll the six cases of PRES had different inducing causes such as acute hypertension,preeclampsia or eclampsia, taking immunosuppressive agents or steroids.and their clinical symptoms were characterized by sudden occurrence of headache,eclampsia or seizure of epilepsy,altered melltal status,visual disturbances.Clinical symptoms were died out in about one week after prompt and appropriate treatments for high blood pressure.or removal of precipitating factors,or treatment for epileptic seizures or status epilepticus.MRI and CT scanning demonstrated multifocal subcortical white lesions in bilateral parieto-occipital lobes (six cases), bilateralfrontal lobes (two cases),bilateral post temporal lobes (two cases) and left cerebellum (one cases).and cortical involvement (two cases).All lesions appeared unenhanced with gadolinium enhancement. FoHow-up by MRI showed decreased abnormal signs and small infarct foci were left in the cortex-subeortex of one case.ConclusionsPRES is a clinico-neuroradiologieal transient condition, usually benign and reversible in nature.Completely clinical and radiographic recovery Can be achieved with prompt antihypertensive treatment or removal of precipitating factors and supportive care,but delayed diagnosis and therapy Can result in cerebral infarct with neurological sequelae.

Objective To detect specific cell killing effect of radiation combined with horseradish peroxidase (HRP)/indole-3-acetic (IAA) suicide gene therapy controlled by a novel radio-inducible and cancer-specific chimeric gene promoter in lung cancer. Methods We constructed a plasmid expressing HRP enzyme under the control of chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 CArG elements, a plasmid expressing HRP enzyme under the control of hTERT promoter carrying single CArG element, and two control plasmids, which named pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc, respectively. After radiation, the proliferation inhibition and apoptosis induction effect of each type of plasmid in lung cancer cells (A549, SPC-A1) and normal lung cells (hEL) was detected by cell counting and Annexin V-FITC staining. The change of radiosensitivity of lung cancer cells with plasmid system was also detected by clonogenic assays. Results After a single dose radiation of 6 Gy,the average proliferation inhibition rates of pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc systems were 72. 92% ,40.60% , 51.00% and 25.19% (F= 67.31 , P< 0.01) in A549 cells ,64.63%,30.02%,48.23% and 23.16% (F=64.94, P< 0.01) in SPC-A1 cells, and 20.81%,18.05%, 44.20% and 18.32% (F=52. 19,P<0.01) in normal hEL cells, respectively. The average early apoptosis rates of these four plasmid systems were 36. 63%, 22. 30%, 24. 33% and 12. 53% (F =50. 99,P <0. 01) in A549 cells, 33.73%, 17. 37%, 22. 43% and 11.20% (F = 20. 76, P < 0. 01) in SPC-A1 cells, and 13.53 %, 12. 5%, 21.93% and 12. 16% (F = 15. 08, P < 0. 01) in normal hEL cells,respectively. The sensitizing enhancement ratios of the four plasmid systems were 3.45, 2. 29, 3.05 and 1.21 in A549 cells, while 2. 68, 2. 15, 3.05 and 1.21 in SPC-A1 cells, respectively. Conclusions The new suicide gene system controlled by chimeric promoter may provide a novel therapeutic modality for lung cancer.