OBJECTIVE:To establish a method for the concentration determination of catecholamines in human urine. METH-ODS:After extracted by aluminum oxide,using 3,4-dihydroxybenzylamine as internal standard,HPLC-electrochemistry was per-formed on the column of Shim-Pack C18 with mixed solution [every 500 ml mixed solution containing ion-pairing reagents(octane sulfonate) 0.5 mmol,methanol 10 ml,0.06 mmol/L Di-n-butylamine 50 μl,sodium acetate anhydrous 2.0 g,citric acid 5.5 g, EDTA-Na2 60 mg and sodium chloride 584.4 mg,pH=3.5] at a flow rate of 0.8 ml/min,column temperature was 30 ℃,detection voltage was 750 mV,and injection volume was 40 μl. RESULTS:The linear ranges were 3.125-100 ng/ml for both norepinephrine (r=0.998 5)and epinephrine(r=0.999 2),lower limits of quantitation were 3.125 ng/ml;RSDs of within days and daytime were lower than 10%,method recoveries were 99.2%-101.6% and 98.9%-100.5%,extraction recoveries were 63.8%-66.8% and 60.5%-62.6%. CONCLUSIONS:The method shows strong specificity,high sensitivity and low detection cost,and can be used for the concentration determination of catecholamines in human urine.

Objective: To investigate the expression of human telomerase reverse transcriptase (hTERT)in benign and malignant gastric diseases and its clinical significance. Methods: The expression of hTERT in 126 gastric tissue specimens obtained by surgery and endoscopy were studied by SP immunohistochemical method. The specimens included 10 cases of nomal mucosa,26 of chronic gastritis, 22 of benign ulcer,5 of leiomyoma, 3 of hyperplastic polyps, 50 of gastric cancer, 2 of maligmant lymphoma and 8 of sarcoma. In precancerous lesions,there was 54 cases of atypical hyperplasia,including 18 low grade, 25 of intermediate and 11 high grade. The positive expression rates and relations of clinical characters were analyzed. Results: hTERT was not or seldom expressed in normal and benign gastric diseases, while it was highly expressed in malignant lesions.There was a significant difference between them(P

Objective To investigate the effect of Salvia miltiorrhiza extracts on morphine physical dependence in mice. Methods The physical dependence model was induced by repeated sc morphine daily in mice and then withdrawal symptom was induced by ip naloxone. Different doses of S. miltiorrhiza extracts were administrated by ip during or after the induction phase of morphine dependence, and effects of S. miltiorrhiza extracts both by preventive and acute administration on naloxone-precipiated withdrawal symptom in mice were investigated. The potential physical dependence of S. miltiorrhiza extracts was studied. Results Compared to morphine model group, preventive administration of S. miltiorrhiza extracts (100—200 mg/kg) could reduce the number of naloxone-induced withdrawal jumps (P

ObjectiveTo investigate the expression and clinical significance of CD40 in the spleen of cirrhotic portal hypertension. MethodsExpression of CD40 was determined with S-P immunohistochemistry in spleen specimens from 50 cases of cirrhotic portal hypertension and 15 healthy individuals. ResultsThe CD40 positive rates in normal spleen and cirrhotic spleen were 86.7% and 36.0%, respectively. There was a significant difference between the 2 groups (P＜0.05). There was a negative correlation between the expression of CD40 and Child grades of liver function and cirrhotic splenic types(P＜0. 05). Conclusion CD40 might reflect the changes of splenic immune function,which might be one of more exact clinical examination indexes of splenic immune function.

Objective To discuss the diagnosis and surgical treatment of insulinoma. Methods The clinical data of 32 patients with insulinoma were analyzed retrospectively. Of the 32 patients, 19 patients were performed with simple tumor enucleation, 10 patients with pancreatic body and tail resection, 3 patients with pancreaticoduodenectomy. Results In this study, 20 cases were diagnosed and localized the tumor by type-B abdominal ultrasonic and enhanced spiral CT before operation and 12 cases by intraoperative type-B ultrasonic. Within all patients,9 tumors located in the head of pancreas, 15 tumors in the neck , 8 tumors in the tail. The clinical symptoms in 32 cases disappeared after operation with 2 cases followed by pancreatic leakage. Of all 32 cases with insulinoma,28 tumors were diagnosed as benign and the other 4 as malignant. Conclusions The mainly clinical manifestation of insulinoma is typical whipple triad. Preoperative qualitative diagnosis is practicable according to the clinical symptom and laboratory examination. Combined imaging examinations might be helpful in improving the positive test rates. Intraoperation exploration and ultrasonography are the chief methods for the tumor localization. Site, size, number and depth of the tumor are the key points determining the surgical method style.

Objective To evaluate the indications and clinical efficacy of the different surgical methods for lumbosacral tuberculosis.Methods 112 patients suffering from lumbosacral tuberculosis from January 1998 to October 2011 were reviewed retrospectively.Based on American Spinal Injury Association (ASIA) grading system,31 case was classified as grade C,32 as grade D and 49 as grade E.The affected locations were L5,S1 in 66 cases,L4-S1 in 32,and L5-S2in 14 cases.41 cases (posterior group) underwent posterior pedicle screw instrumentation,debridement and allograft; and 38 cases (anterior group) underwent one-stage anterolateral debridement plus allograft and internal fixation.33 (posterior-anterior group) cases experienced posterior pedicle screw instrumentation and anterior debridement and allograft.All cases underwent routine support and anti-tuberculosis treatment before and after operation.The change of erythrocyte sedimentation rate (ESR),signs of tuberculosis activity,graft fusion,neurological recovery and correction of deformity were evaluated in follow-up and compared among three groups.Results The average operating time in posterior,anterior and posterior-anterior group was (150±22) m in,(140± 18) min,(180± 1 8) min respectively.ESR and C-reactive protein (CRP) were recovered to normal 3 months post surgery.The quality of life total score of lumbosacral tuberculosis patients was very low (55.54 ± 9.23).After surgery and drug treatment,the total score and each dimension scores of the SF-36 were significantly improved.Postoperative ASIA classification and 1umbosacral angle were significantly improved.Tuberculosis recurrence occurred in three cases,fixation loosening in two cases,fixation fracture in one case,and all above cases were cured after revision surgery.The others all were cured and bone graft fusion was determined.Conclusion The quality of life of lumbosacral tuberculosis patients were impacted seriously,and which could be significantly improved with surgical intervention.According to the patient MRI,CT characteristics,surgical approach selection was based on lesion location.Effective internal fixation was based on the extent of damage in bone structure and anatomical characteristics.Following above principles,satisfactory clinical results could be achieved.

Objective: To investigate the role of expression of Caspase-9 in megalosplenia of cirrhotic portal hypertension. Methods: Using SP immunohistochemistry method to examine theexpression of Caspase-9 in the specimens of 50 cases of megalosplenia with cirrhotic portal hypertension and 15 cases of traumatic spleen of normal man. Results: The Caspase-9 positive rates in the traumatic spleen and megaloplenia were 26.7% and 76.0%, respectively. There was a significant difference between the groups(P

Objective To investigate clinical result of hepatectomy and porta-enterostomy in the treatment of Bismuth type Ⅳ hilar cholangiocarcinoma.Methods Nine patients with Bismuth type Ⅳ hilar cholangiocarcinoma underwent accurate hilar resection(portal parecnchyma resection including about Ⅰ cm of the hilar part of the segments 5 and 4b and caudate lobe beyond the tumor),and the biliary drainage was reconstructed by Roux-en-Y portal parecnchyma-jejunum lpop anastomosis.None of the biliary radicals had to be ligated and all of them were drained into thus constructed"biliary pool".Results Hilar resection was successfully performed in all cases,and there was no postoperative mortality.Aspartate transaminase and alanine transaminase and serum bilirubin decreased evidently four weeks later.Three patients presented postoperative complications.One patient developed a transient anastomotic leakage,while one patient developed self-limiting hemobilia,wound infection occurred in one patient.All three patients were treated conservatively and recovered.The mean Karnofsky performance score was 86,with which they could carry on normal activity with minor symptoms of disease.Two patient died after 9 months and 17 months of extensive metastasis and intrahepatic metastasis respectively.The remaining seven patients are alive by a mean followup of 24.9 months after surgery without any signs of recurrence.Conclusions With accurate hilar resection and portal parecnchyma-to-enterostomy,the patients considerably benefit from the preservation of liver parenchyma and patent biliary drainage and radical resection.So the Hew technique prolongs the survival time and enhances the quality of life of the patients.

Objective To investigate the therapeutic effect of the meso-cavo-atrial shunt (MCAS) in the treatment of combined Budd-Chiari syndrome (BCS). Methods The clinical data of 17 cases of combined BCS with all or bilateral hepatic vein occlusion and long range occlusion or obstruction of inferior vena cava (IVC) were admitted to the Qilu Hospital from February 2000 to May 2004. All patients were treated by MCAS with artificial blood vessels. The pre- and postoperative clinical symptoms, the IVC and portal venous (PV) pressures, the incidence of postoperative complications and the patency rate of the artificial vessels were analyzed. The survival of patients was analyzed using the Kaplan-Meier analysis, and the data were analyzed using the chi-square test and t test. Results No patient died during the perioperative period, and the symptoms of 15 patients disappeared or were relieved after operation, with a significant difference compared with those before operation (χ2 =9.78, P <0. 05 ). Three patients had complications after the operation. The postoperative PV and IVC pressures were decreased by 1.2 cm H2O (1 cm H2O =0.098 kPa) and 18.5 cm H2O, respectively. There were significant differences in the decrease of IVC and PV pressures ( t = 2.38, 3.06, P < 0.05 ). The 1-, 3-, 5-year survival rates were 16/17, 15/17 and 14/17, respectively, and the 5-year patency rate of the artificial vessels was 14/17.Conclusions MCAS can simultaneously relieve IVC and PV hypertension for patients with combined BCS. The postoperative complication rate was decreased, the 5-year survival rate and the patency rate of the artificial vessels were improved after the treatment, so MCAS is an optional surgical method for treating combined BCS.

ObjectiveTo detect the mRNA and protein expressions of Bmi-1 in hepatocellular carcinoma (HCC) tissue,pericarcinomatous tissue,portal vein tumor thrombus and normal liver tissue,and to investiage the significance of Bmi-1 in the genesis and progression of HCC.MethodsForty tissues of HCC were collected from the Tongji Hospital of Huazhong University of Science and Technology from January 2005 to December 2009.The mRNA and protein expressions of Bmi-1 in the HCC tissues (40 cases),pericarcinomatous tissues (40 cases),portal vein tumor thrombus ( 11 cases) and normal liver tissues ( 10 cases) were detected by immunohistochemistry,Western blot and real-time polymerase chain reaction.The relationship between the expressions of Bmi-1 and the clinicopathological factors was analyzed.Differences in each group were compared by using the Nemenyi test or Dunnet t test,and the relationship between the clinicopathological factors and the protein expression in the HCC tissues was analyzed by the chi-square test or Fisher exact probability.The survival curve was drawn by the Kaplan-Meier method,and the survival rate was analyzed by the Log-rank test.Results The median relative mRNA expressions of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 0.96,2.60,7.51 and 29.95,respectively.The results of immunohistochemistry showed that the high protein expression rates of Bmi-1 in the normal liver tissues,pericarcinomatous tissues,HCC tissues and portal vein tumor thrombus were 10.0％,20.0％,67.5％ and 100.0％,respectively.The high protein expression rates of Bmi-1 in the HCC tissues and portal vein tumor thrombus were significantly higher than those in the normal liver tissues and pericarcinomatous tissues (x2 =17.25,22.77;22.04,23.95,P ＜ 0.05 ). High protein expression of Bmi-1 was also detected in 11 cases of HCC tissues with portal vein tumor thrombus.The results of western blot were consistent with those of the immunohistochemistry.The mRNA and protein expressions of Bmi-1 were correlated with Edmondson grade and portal vein metastasis ( x2 =5.572,P ＜ 0.05 ),whereas they were irrelevant to the tumor size,serum levels of α-fetoprotein and hepatitis B surface antigen ( x2 =0.000,0.019,0.663,P ＞0.05).Patients with high expression of Bmi-1 had poor prognosis.ConclusionsBmi-1 is correlated with the genesis and progression of HCC as well as the formation of portal vein tumor thrombosis.Patients with high Bmi-1 expression have poorer prognosis when compared with those with low Bmi-1 expression.