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 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.

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 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 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 value of MRCP in conjunction with CT or MRI contrast-enhanced scan in locating and qualitative diagnosis of biliary obstruction.Methods 954 patients with biliary obstruction confirmed by surgical pathology and clini-cal follow up underwent MRCP scans in our hospital.Contrast-enhanced CT scans in 87 patients,contrast-enhanced MRI scans in 52 and both CT and MRI enhancement in 37 were performed.Results The accuracy of location of biliary obstruction with MRCP was 100%,the accuracy of quality of biliary obstruction with MRCP combined with CT or MRI dynamic contrast-enhanced scan was 96%.Conclusion MRCP combined with CT or MRI dynamic contrast-enhanced scans has an important clinical value for the locali-zation and qualitative diagnosis of biliary obstruction.

BACKGROUND: Currently, the researches on knee plaster immobilization paralysis animal models are popular in the world, but there are some insufficiencies with the knee paralysis animals, for example, poor animal selection, inappropriate plaster selection and pure gypsum instability, which affect the subsequent results.
OBJECTIVE: To establish a rabbit model of knee paralysis by the knee fixation with plaster casts and wire.
METHODS: A total of 20 New Zealand white rabbits were randomly divided into two groups. Models of right knee paralysis were established and fixed by plaster casts and wire. Normal controls were considered as the control group. At 8 weeks after fixation, right knee and pathologic histology were observed in the right knee.
RESULTS AND CONCLUSION: The knee joints were translucent, smooth, and shiny, without the cartilage defect in the control group, and the cartilaginous elasticity and rigidity were good, and the arrangement of chondrocytes was normal. There were no inflammatory infiltrations in the articular capsule of the normal rabbits. In the model group, the knee joint was matt and opaque. Articular cartilage became thin, and showed poor elasticity. The cartilages were degenerated. Articular surface was rough, and had defects. There were smal amount of joint fluid or no joint fluid. Cartilage cel s shrank or disappeared, and the number of cartilage cel s reduced with degeneration and necrosis. Subchondral bone became sclerosis with trabecular bone of hyperplasia. The arrangement of cartilage cel s was disordered, with hyalinization and synovial vil ous hypertrophy. Cel clustering phenomenon was seen. There were a large number of lymphocytes, plasma cel s and neutrophil infiltration in cartilages. The typical pathological changes of the knee paralysis present in the rabbit cartilage. These results demonstrate that the rabbit model of knee paralysis can be successful y established by using fixation method with plaster casts and wire, which has the advantages of simple operation, strong fixation and no injury.