Objective To investigate the expressions and significance of p21 protein and K-ras protein in congenital biliary dilatation(CBD),and their relationship.Methods Surgical specimens of CBD were obtained during operation on 36 children from Jan.2000 to Sep.2007.Nine normal bile ducts specimens from miscarried fetus and children's corpse.The specimens were fixed in 40 g?L-1 formalin and embeded by paraffin,slice-fixed.SP immunohistochemical method was used to detect the expressions of p21 protein and K-ras protein.Results The positive rate of K-ras protein was 13.8% in CBD,which was significantly higher than that in normal bile duct tissues.The positive rate of p21 protein was 55.6% in CBD,and 100% in normal bile duct tissues.p21 protein positive expression rate in CBD was significantly lower than that in normal bile duct tissues(P

Objective To prepare a 10-hydroxycamptothecin (10-HCPT) loaded folate-receptor targeted phase-change contrast agent (FR-HCPT-PNPCA),and to study the general characteristics including drug loading,phase changing and targeting capability in vitro.Methods Using a method of two-step emulsification,the phase-change nanoparticles loading anticancer drug (10-HCPT) with lipids shell and liquid pefluorocarbon core were prepared.The entrapment efficiency and the drug-loading amounts were studied by high performance liquid chromatography,and the phase transition of the nanoparticles after heating was observed.The targeting ability was evaluated on liver cancer cell line 7721 in vitro.Results The FR-HCPT-PNPCA,with a drug encapsulation rate of about 70.42 ％ and drug loading amounts of about 20.05 ％,was prepared successfully.When being heated to 70℃,obvious phase changing and microbubbles generating could be observed under microscope.In addition,a large amount of FR-HCPT-PNPCA particles could adhere specifically around the 7721 cells.Conclusion The prepared FR-HCPT-PNPCA,which has a stable characteristic and high performance of drug loading and tumor targeting,is expected to become a promising multifunctional molecular ultrasound probe for diagnosis and treatment of tumor.

Objective To construct the luciferase reporter gene vector of cell division cycle 2 (Cdc2) gene promoter and determine its transcriptional activity. Methods Primers were designed based on human Cdc2 promoter sequence from UCSC software. Then Cdc2 promoter from human genome DNA was replicated. After pGL3-Basic vector and Cdc2 promoter were digested with restriction enzymes SacⅠand XhoⅠseparately, Cdc2 promoter was inserted into pGL3-Basic vector. The recombinant plasmid named pGL3-Cdc2-promoter was transiently co-transfected into U2OS cells with control vector pRL-SV40, and then the activity of dual luciferase was detected. Results pGL3-Cdc2-promoter was constructed successfully. The restriction analysis and sequencing proved the entirely correct sequencing results. The luciferase activity was higher in pGL3-Cdc2-promoter/pRL-SV40 group than that of pGL3-Basic/pRL-SV40 group (1.591 5±0.199 8 vs 0.049 9±0.010 4). Conclusion pGL3-Cdc2-promoter can be transcribed and activated in U2OS cells. This study provided an important basis for screening and evaluation of anticancer drugs.

Objective To review the clinical,histological and diagnostic aspects of 12 documented cases of nodular regenerative hyperplasia of the liver(NRHL),to make this condition be understood and dealt with better. Method Twelve NRHL cases were diagnosed based on liver biopsy from 300 portal hypertension patients who had been underwent splenectomy.Imaging studies were performed as part of the diagnostic evaluation.Clinical manifestation and biochemical tests were recorded at the time of diagnosis.Management and prognosis were also reviewed.Results Most patients were complicated with autoimmune disease,6 cases were diagnosed systemic lupus erythematosus and 1 was Crohn's disease and 1 suspected ulcerative colitis.Six cases were treated by prednisone and 3 cases by immunosuppressant.Eleven cases had suffered from portal hypertension.All cases had no history of viral hepatitis.Biochemical tests showed mild increase of liver enzyme and relative normal synthetic liver function.The histological finding was nodular in the hepatic parenehyma,with mild periportal fibrosis,intraportal lymphocytic infiltration,narrow and obstruction of branch of portal vein,and lack of hepatocyte necrosis.All cases were diagnosed liver cirrhosis and portal hypertension before operation.Management was directed to portal hypertension and varices bleeding with satisfactory results.Most of them keep a stable condition during the follow-up. Conclusion The NRHL was uncommon and its cause and pathogenesis was unclear,may be related with immune and hepatic blood circulation disorder.It should be considered in patients with unexplained portal hypertension and distinguished it from liver cirrhosis.Liver biopsy confirms the diagnosis.Management directed to portal hypertension may improve clinical condition.

Objective To establish the high performance liquid chromatography(HPLC) fingerprint of wuzi yanzong pills. Methods HPLC was performed on Agilent Extend C18 column (250 mmí4. 6 mm,5 μm) with a gradient elution system using acetonitrile:methanol (101)-0. 4% phosphoric acid as the mobile phase. The column temperature was set at 30 ℃ and the flow rate was 1. 0 mL·min-1 . The eluate was detected at the wavelength of 254 nm. Chromatographic peaks were identified by LC-MS method. Results Nine common peaks in wuzi yanzong pill samples were identified by comparing their LC-MS data with those of reference compounds and related reference reports. The HPLC fingerprint of wuzi yanzong pills was finally developed based on the analysis of sixteen batches of samples and their similarities were above 0. 93. Conclusion This method has high precision,stability and repeatability. This study could be used for overall quality assessment of wuzi yanzong pills.

Objective To explore the effects of hyperglycemia and oxidized low density lipoprotein (ox-LDL) on the differentiation of macrophage derived THP-1 monocytes. Methods THP-1 human monocytic leukemia cell line was cultured in vitro, and the differentiation of THP-1 cells into macrophages was induced by phorbol esters. The macrophages were then incubated with the absence of D-glucose and ox-LDL (control group), 30 mmol/L D-glucose (hyperglycemia group), 100 μg/mL ox-LDL (ox-LDL group) or 30 mmol/L D-glucose and 100 μg/mL ox-LDL(G-ox-LDL group) for 24 h. High performance liquid chromatography was used for qualitative and quantitative analysis of intracellular cholesterol and cholesteryl esters. Both light microscope with red oil O staining technique and transmission electron microscope were employed to observe the morphology of treated and control THP-1 cells. Results A large number of intracellular red oil O stained granules and lipid vacuoles were observed in ox-LDL group and G-ox-LDL group, the contents of total cholesterol and cholesteryl esters were significantly higher than those of control group (P<0.05), and the contents of cholesteryl esters were higher than 50% of total cholesterol in both groups. However, only a few intracellular red oil O stained granules and lipid vacuoles were observed in control group and hyperglycemia group, there was no significant difference in the contents of total cholesterol and choleateryl esters between control group and hyperglycemia group (P>0.05), and the contents of cholesteryl esters were less than 50% of total cholesterol in both groups. Conclusion Foam cells form when THP-1 cells are incubated with ox-LDL, while hyperglycemia alone can not convert THP-1 cells to foam cells, indicating that ox-LDL is necessary for the macrophages derived THP-1 monocytes to turn into foam cells.

Objective To discuss the value of applying psychological therapy for patients with cerebral apoplexy in recovery.Methods In recent years,65 patients with cerebral apoplexy admitted in our hospital were randomly selected and divided into observation group and control group.Patients in the control group only received the conventional treatment and those in the observation group received psychological treatment on the basis of the traditional therapy.Psychological characteristics,activity of daily living and the improvement of nerve function were compared between two groups before and after treatment.Results After treatment,mental improvement,FMA,MBI in the observation group were significantly better than those of control group (P＜0.05).Conclusion To improve the prognosis of patients with cerebral apoplexy effect,physical and mental development should be equally emphasized.Psychological intervention has become a key for stroke rehabilitation treatment,which significantly improves the treatment compliance,rehabilitation and the quality of life in patients.

Objective To observe the effects of hypnosis therapy on rehabilitation of patients with spinal cord injury.Methods The data were randomly selected from 66 cases of patients with spinal cord injury treated in our hospital from November 2011-November 2012.According to the treatment,the patients were divided into research group and control group.Control patients were given routine therapy,and patients in the research group were given hypnosis therapy,we observed and compared the recovery of two groups of patients after treatment.Results After treatment,the spinal cord function score (SCIM) of patients in the research group was better than control group,the difference was statistically significant (P ＜ 0.05) After treatment,the depression and anxiety scale (HAMD,HAMA) of patients in the research group was better than control group,the difference had statistical significance (P ＜ 0.05) The quality of life after treatment of research group patients (WHOQOL BREF) was better than control group,the difference had statistical significance (P ＜ 0.05).Conclusion Hypnosis therapy helps to improve depression and anxiety in patients with spinal cord injury,improve patients with spinal cord function independently,should be widely used in clinical treatment and promotion.

Objective To analyze the therapeutic effect of acute stroke dysphagia using rehabilitation therapy.Methods A total of 94 cases diagnosed with dysphagia after acute stroke were randomly selected from January 2012 to May 2013 in our hospital.The patients were equally divided into control group and research group.The control group received the conventional support treatment and the research group received comprehensive rehabilitation treatment.The results of swallowing dysfunction recovery of two groups were compared.Results After the treatment for 3 weeks,swallowing disorder degree score and swallowing function ratings were improved in both groups and the research group's scores were significantly higher than those of the control group (P ＜0.05) Conclusion Application of comprehensive rehabilitation treatment for dysphagia in patients with acute stroke has significant curative effect and is worth promoting.

Objective To investigate the incidence of postoperative hepatic metastasis,clinicolpathological characteristics and the prognosis for pancreatic cancer. Methods Totally 83 cases with pancreatic cancer admitted in our hospital during January 2007 to September 2012 was retro?spectively analyzed according to clinicolpathological data. Results Postoperative liver metastasis occurred in 31 cases with a metastatic rate of 37.3%. The size(χ2=9.606;P=0.002),vascular invasion(χ2=4.794,P=0.029)and UICC stage(χ2=5.318,P=0.021)were correlated with he?patic metastasis. Univariate analysis revealed the poor prognosis in pancreatic cancer patient with hepatic metastasis(χ2=9.967,P=0.002). Cox re?gression analysis revealed hepatic metastasis as an independent prognostic factor(P=0.001). Conclusion Pancreatic cancer has a high possibility of hepatic metastasis. Postoperative hepatic metastasis was one of the independent factors for the prognosis of pancreatic cancer. Tumor size,vascular invasion and UICC stages were risk factors for postoperative liver metastasis of pancreatic cancer.