Objective: To establish a method for the determination of norcantharidin in norcantharidin in situ gel .Methods:An optimal HPLC method was set up and an Agilent ZORBAX SB-C18 column (150 mm ×4.6 mm, 5μm) was adopted.The mobile phase was acetonitrile-phosphate buffer solution(1∶9, adjusting pH to 3.1 with phosphorjc acid).The flow rate was 0.8 ml· min-1 and the column temperature was 25℃.The detection wavelength was set at 210 nm and the injection volume was 20μl.Results:Norcanthari-din had a good linear relationship within the range of 0.02-1.00 mg· ml-1 (r=0.999 9).The average recovery was 97.5%and RSD was 0.98%(n=9).Conclusion:The method is accurate, simple and reproducible, which can be used for the determination of nor-cantharidin in norcantharidin in situ gel .

Objective To study the inhibitory effects of adenovirus-mediated fusion gene system driven by KDR promoter(AdKDR-CDglyTK) on angiogenesis and growth of pancreatic cancer.Methods The nude mice model was reproduced bearing pancreatic cancer cell lines Capan-2.Twenty nude mice were randomly and equally divided into four groups: AdKDR-CDglyTK and 5-FC/GCV were injected to the animals of group Ⅰ;group Ⅱ received 5-FC/GCV injection,group Ⅲ received AdKDR-CDglyTK injection,and group Ⅳ as control,received neither AdKDR-CDglyTK nor 5-FC/GCV.AdKDR-CDglyTK was injected directly into the tumor,while 5-FC/GCV was given by intraperitoneal injection.The treatment efficiency in each group was observed and the tumor microvessel density(MVD) was analyzed.RT-PCR was employed to examine the expression of CDglyTK in tumors.Results Tumor growth was dramatically inhibited in group Ⅰ,while no significant difference was found in group Ⅱ,group Ⅲ and group Ⅳ.The MVD in the four groups were 2.08?0.79,10.01?0.77,9.91?0.63 and 10.39?1.35,respectively(F=93.29,P=0.00).The MVD decreased significantly in group Ⅰ compared to the other three groups(P0.05).RT-PCR showed that a 2.4kB fragment had been amplified in the tumor tissues of groupⅠand group Ⅲ,but not in groupⅡand group Ⅳ.Conclusion AdKDR-CDglyTK with 5-FC and GCV can significantly inhibit the angiogenesis and growth of implanted pancreatic cancer in nude mice.

Objective To compare the diagnostic efficiency in showing the responsible blood vessels for neurovascular compression in patients with trigeminal neuralgia by 3D‐FIESTA‐C and 3D‐TOF‐MRA sequences .Methods The imaging data of 60 patients with primary trigeminal neuralgia were analyzed retrospectively .After MRI examination ,all of the patients underwent micro‐vascular de‐compression (MVD) .3D‐TOF‐MRA and 3D‐FIESTA‐C sequences were performed to evaluate the three‐dimensional relationship be‐tween trigeminal nerve and blood vessels through the original and reconstructed image .The intraoperative endoscopic findings were set as the gold standard comparing to the manifestations of 3D‐TOF‐MRA and 3D‐FIESTA‐C .Results The sensitivities of 3D‐TOF‐MRA and 3D‐FIESTA‐C for the diagnosis of the existence of responsible vessels were 85 .7% ,89 .3% ,the specificities were 75 .0% , 100% ,and the accuracies were 85 .0% ,90 .0% ,respectively (P=1 .000) .Furthermore ,the sensitivities of 3D‐TOF‐MRA and 3D‐FIESTA‐C for the diagnosis of the existence of responsible arteries were 94 .1% ,88 .2% (P=0 .244) ,while the sensitivities of the responsible veins were 0 .00% and 88 .2% (P=0 .009) .Conclusion Both the 3D‐FIESTA‐C and 3D‐TOF‐MRA sequences can accurately deter‐mine the existence of responsible vessels in trigeminal neuralgia before surgery .3D‐FIESTA‐C sequence is superior to 3D‐TOF‐MRA for presenting the responsible veins ,which can be used as a supplemental diagnostic tool before operation .

Objective To investigate the expressions and clinical pathological significance of EpCAM and the β-catenin pathway in colon cancer,and the correlation between EpCAM andβ-catenin.Methods Immunohistochemistry was used to detecte the expressions of EpCAM and β-catenin proteins in colon cancer tissues and corresponding adjacent tissues of 70 cases of colon cancer patients,the clinical and pathological features of colon cancer and their relationship were retrospective analyzed.Results ①EpCAM and β-catenin protein expressions in colon cancer tissue was 52 cases (74.3％) and 55 (78.6％) positive respectively; EpCAM and β-catenin proteins in cancer adjacent tissues in 13 cases( 15.7％ ) and 9 cases ( 12.9％ ) were positive,the differences were statistically significant.②the EpCAM expression is positively correlated with the β-catenin expression ( r =0.616,P ＜ 0.01 ).③The expressions of EpCAM and β-catenin correlated with the tumor differentiation,invasion depth,lymph node metastasis and TNM staging related ( P ＜ 0.05).Conclusion EpCAM and β-catenin pathways in colon cancer positively correlate,and closely correlate with colon cancer.The expression level of EpCAM and β-catenin can be used as a reference for the colon disease course and healing.

Objective EpCam and Wnt/ β-catenin pathway in colon carcinoma and its clinic-pathological significance of the distribution,studying the relationship between EpCam and Wnt / β-catenin.Methods Retrospective analysis detected by immunohistochemistry 60 cases of colon cancer,20 cases of adjacent atypical hyperplasia,60 normal colon tissue EpCam and Wnt / β-catenin protein expression.Results (1)normal colon tissue,cancer tissue and cancer tissue showed positive expression EpCam clear upward trend,were 23.5％,62.3％,96.5％ ; with normal colonic mucosa to cancer transformation,β-catenin in the membrane expression of the positive rate decreased,while the cytoplasm is followed by increased expression rate in poorly differentiated carcinoma or even nuclear expression,EpCam strong positive expression of Wnt / β-catenin cytoplasmic-positive rate of histological type,depth of invasion,lymph node metastasis; (2)the EpCam with the Wnt / β-catenin expression showed a positive correlation (r =0.653,P ＜0.05) ;(3)high expression EpCam and Wnt / β-catenin in patients with colon significant increase in cancer recurrence rate and 5-year survival rate was significantly reduced.Conclusion EpCam and Wnt / β-catenin pathway in colon cancer positively is correlated,EpCam and Wnt / β-catenin is connected with high expression and tumor invasion,metastasis and prognosis.

Objective To explore the down-regulation of advanced receptor for advanced glycation end products(RAGE)on expression of high mobility group protein B1(HMGB1)in glioma cells line and the volume change of transplanted tumor in nude mice. Methods HMGB1 expression in glioma LN229 cells line (divided into a control group and a study group) was observed by immunohistochemical assay and Western blot. The control group received normal saline,whereas the study group received RAGE receptor blocking agent FPS-ZM1. Expression of HMGB1 protein was detected by the same methods. The difference of the expression was examined by independent sample t test. 30 Nu/Nu nude mice were randomly divided into two groups;the above two kinds cell lines were injected into the same area of the left back of nude mice. Six weeks after injection ,the volume size was measured six times ,and the variance of repeated measurement data was used to analyze the difference of the volume change. Results HMGB1 protein was mainly expressed in the cytoplasm and nucleus. As compared with the control group,HMGB1 protein expression levels were decreased in the study group(P < 0.05),the growth rate of transplanted tumor in nude mice was significantly faster in the control group than in the study group ,the difference was statistically significant(P < 0.05). Conclusions The growth and invasion of HMGB1 protein may be involved in glioma by RAGE receptor. RAGE receptor blocker FPS-ZM1 can significantly reduce the expression of HMGB1 protein and inhibit the growth of transplanted tumor volume. It is expected to be used for the research on glioma cell apoptosis.

Objective To investigate the feasibility, safety and effectiveness of stent-assisted coil embolization for the treatment of intracranial wide-necked aneurysms. Methods The clinical and imaging data of the patients with intracranial wide-necked aneurysm treated with stent-assisted coil embolization were analyzed retrospectively. Results A total of 200 patients with 205 aneurysms were enrolled. The mortality was 1. 5% and the disability rate was 1. 0% at discharge. One hundred seventy-seven patients were followed up for 16-51 months. The modified Rankin Scale scores: 0 in 174 cases, 2 in 2 cases, 4 in 1 case. Eleven patients (5. 5% ) had perioperative complications, including intraoperative bleeding in 3 cases, postoperative bleeding in 3 cases, postoperative cerebral infarction in 2 cases, coil protrusion in 2 cases, and postoperative epileptic seizure in 1 case. Univariate analysis showed that there were significant differences in the proportions of male patients (9. 1% vs. 5. 3% ; χ2 = 4. 42, P = 0. 026), hypertension (54. 5% vs. 23. 3% ; χ2 = 5. 42, P = 0. 03) and stent prior to coil implantation (54. 5% vs. 85. 1% ; χ2 = 3. 54, P =0. 021) between the complication group and the noncomplication group. Multivariate logistic regression analysis showed that the pre-stenting was an independent protective factor for surgery-related complications (odds ratio [OR] 0. 208, 95% confidence interval [CI] 0. 055-0. 791; P = 0. 021), and hypertension was an independent risk factor for surgery-related complications (OR 4. 380, 95% CI 1. 170-16. 399; P = 0. 028). The imaging follow-up of 167 aneurysms was obtained, including 26 recurrent aneurysms (15. 6% ). Univariate analysis showed that there was significant difference in the aneurysm site (anterior circulation aneurysms: 73. 1% vs. 89. 1% ; posterior circulation aneurysms: 26. 9% vs. 10. 6% ; χ2 = 5. 09, P = 0. 033) and size (giant aneurysms: 7. 7% vs. 0. 7% ; large artery aneurysm: 65. 4% vs. 29. 1% ; small aneurysms:26. 9% vs. 70. 2% ; χ2 = 20. 77, P < 0. 001) between the recurrence group and the nonrecurrence group. Multivariate logistic regression analysis showed that large aneurysms (OR 6. 057, 95% CI 2. 296-5. 983; P <0. 001), giant aneurysms (OR 25. 260, 95% CI 1. 903- 335. 267; P = 0. 014 ), and posterior circulation aneurysms ( OR 3. 184, 95% CI 1. 028- 9. 857; P = 0. 045 ) were the independent risk factors of postoperative recurrence. Conclusions Stent-assisted coil embolization is one of the effective methods for the treatment of complex wide-neck aneurysms. Hypertension and coils prior to stenting are the independent risk factors for perioperative complications, and larger aneurysm size and aneurysms in the posterior circulation are the independent risk factors for postoperative recurrence.

Objective:To investigate the stability of flurbiprofen axetil lipid microspheres injection combined with 0. 9% sodium chloride injection or 5% dextrose injection, and provide theoretical basis for the clinical application. Methods:The content changes of flurbiprofen axetil in the mixture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chloride injection or 5% dextrose injection were determined in 5 h at 25℃ away from light, and the changes in the appearance and particle size of flurbiprofen axetil lip-id microspheres were investigated. The changes in the appearance and particle size of flurbiprofen axetil lipid microspheres in the mix-ture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chloride injection before and after freezing and thawing were also investigated. Results:The appearance, particle size and content had no significant changes in all mixtures in 5 h at 25 ℃ away from light. The appearance and particle size of flurbiprofen axetil lipid microspheres in the mixture before and after freezing and thawing had no significant changes as well. Conclusion:The mixture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chlo-ride injection or 5% dextrose injection is stable in 5 h away from light.

BACKGROUND: Studies have demonstrated that exogenous neural stem calls (NSCs) could repair nerve and promote recovery of neurofunction following cerebral hemorrhage. However, the influence of internal environment after cerebral hemorrhage on the survival and differentiation of NSCs is a complex and variable process.OBJECTIVE: To observe the survival and differentiation of human embryonic NSCs implanted in rats with cerebra hemorrhage.DESIGN, TIME AND SETTING: Open, immunohistochemistry, experiment was performed at the Laboratory of Molecular Biology, Affiliated Hospital of Luzhou Medical College from May 2007 to April 2008.MATERIALS: A total of 40 female SD rats were provided by the Experimental Animal Institute, Chinese Academy of Medical Sciences. Brain of 8-week aborted fetus was obtained from Department of Gynaecology and Obstetrics, the People's Hospital of Deyang City.METHODS: Cerebral cortex cells of 8-week aborted human fetus were harvested and cultured in vitro to obtain human embryonic NSCs. Cerebral hemorrhage rat models were established via injection of autologous arterial blood in caudate nucleus. Two days after modeling, 5 μL BrdU-labeled human embryonic NSCs suspension was transplanted at four points surrounding hematoma cavity in the rats. After 1 and 2 weeks, rats were sacrificed. Adjacent sections were doubly stained by BrdUImicrotubule-associated protein 2 (MAP-2) and BrdU/glial fibrillary acidic protein (GFAP).MAIN OUTCOME MEASURES: The survival, immigration and differentiation of human embryonic NSOs implanted in rats were observed by immunohistochemistry and immunofluorescance staining.RESULTS: BrdU-positive cells were oval and brown. At 1 and 2 weeks after implantation, BrdU-positive calls survived and migrated, and they migrated more widely at 2 weeks after implantation. At 1 week after implantation, BrdU/MAP-2-positive cells and BrdU/GFAP-positive calls were observed in cerebral tissue sections, and the number of BrdU/MAP-2-positive cells was more than BrdU/GFAP-positive calls; At 2 weeks after implantation, BrdU-positive cells found in choroid plexus and blood capillary were significantly reduced, and BrdU/GFAP-positive calls were more than BrdU/MAP-2- positive cells.CONCLUSION: Implanted human embryonic NSCs can survive and migrate in the hemorrhage region, gradually differentiate into neurons or astrocytes.

Objective To investigate the expression of TROP2 and MMP-9 protein expression in cholangiocarcinomas and their relationship between the pathological behavior and prognosis.Methods A total of 54 patients who were diagnosed with cholangiocarcinoma in the People's Hospital of Binzhou,were retrospectively reviewed.Immunohistochemical staining and Log rank test were used to detect the expression of TROP2 and MMP-9 protein in 54 cases of cholangiocarcinomas and 18 cases of normal bile duct tissues achieved by partial hepatectomy of hepatolithiasis.Results The positive expression rate of TROP2 in cholangiocarcinoma tissues (55.6％) was higher than that of normal bile duct tissues (5.6％).The positive expression rate of MMP-9 in cholangiocarcinoma tissues (51.9％) was higher than that of normal bile duct tissues (11.1％).The differences of the expression of TROP2 and MMP-9 in cholangiocarcinoma of TNM stage,lymph node metastasis and neural invasion were significant(all P ＜ 0.05).There was significant positive correlation between TROP-2 and MMP-9 expression by using spearman correlation analysis (r =0.555,P ＜ 0.001).Survival analysis showed that TROP2 expression was an independent prognostic factor in cholangiocarcinoma.Conclusions TROP2 plays a important role in the development and metastasis of cholangiocarcinoma.Thus,TROP2 may be a prognostic indicator for cholangiocarcinoma.