Objective To observe the effect of limb negative pressure (LNP) on plasma endothelin (ET) and calcitonin gene-related peptide(CGRP) in dogs with arterial occlusive disease (AOD). Methods Fifteen dogs were divided into the treatment group(10 dogs) and the control group (5 dogs). By ligating the branch of femoral artery and inserting spiral wire into the artery of canine left posterior limb, we established a model of extensive arterial occlusive disease of the extremity. The limbs of treatment group were given LNP therapy. Plasma ET and CGRP were determined by radioimmunoassay (RIA). Results Ten days after LNP treatment plasma ET decreased and plasma CGRP increased significantly compared with controls(P

0.05). The satisfied rate of cosmetic results of the former breast were 81.4%, compared with the latter (P

0.05). The upper extremity function in group C was better than that in group H (P

Objective To construct a recombinant bacillus Calmette-Guérin(BCG) vaccines based on different tandem repeats of MUC1 and GM-CSF, rBCG-MVNTR1/4/8-CSF, and to observe the ability of three recombinant BCG vaccines in the inhibition of breast cancer. Methods After MUC1 variable-number tandem repeats (MVNTR1/4/8) were cloned in a stepwise manner, the E. coli-Mycobacteria shuttle expression vector pDE22-MVNTR1/4/8-CSF were constructed by fusing MVNTR1/4/8 and GM-CSF, and then used to transform competent BCG by electroporation after identification by restriction endonuclease digestion analysis and DNA sequencing. A novel breast cancer vaccines, rBCG-MVNTR1/4/8-CSF was constructed. The expression of fused MVNTR1/4/8-CSF protiens was analyzed by SDS-PAGE and Western blot. The ability of rBCG vaccines inhibiting the growth of breast cancer was observed in hu-PBL-SCID mice. The specific T cell responses in mice were assessed by immunohistochemistry. Results The expression of recombinant MVNTR1/4/8-CSF fusion proteins were detected by SDS-PAGE and Western Blot in rBCG-MVNTR1/4/8-CSF vaccines, respectively. Tumor incidence in mice prophylactic immunized with rBCG-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) significantly decreased compared with PBS and BCG-pDE22 control ( 100% ) at 42 days after tumor implantation ( P ＜ 0. 05 ). MCF-7 tumor growth inhibition in rBCG- MVNTR4/8-CSF-immunized mice was more significant than that in controls ( P ＜0. 01 ).The inhibition effect of three rBCG vaccines on breast rumor growth appeared to rise with increase of numbers of the tandem repeats of MUC1. Survival rate was 75% of mice in the rBCG-MVNTR4-CSF-treated group and 87. 5% of mice in the rBCG-MVNTR8-CSF-treated groups at 70 days after tumor implantation; however,survival rate was only 12. 5% in control group( P ＜0. 05). The CD4-positive and CD8-positive lymphocytes were detected only in rBCG-MVNTR4/8-CSF-immunized mice. Conclusions rBCG-MVNTR4/8-CSF immunization inhibits breast cancer growth in mice.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.

Objective Tocheck and evaluate on clinical practice teaching files for providing a foundation of improved clinical teaching levels with enhanced establishment of practice teaching files. Methods The practice teaching files of eighteen clinical departments in a military medical university were selected as the subject and were evenly divided into teaching plans and teaching records as well as teach-ing summary three parts and 15 items. Each item had 2 scores in which quantity had one score and quality has another one. Evaluation results were converted into 100 points system. Results Among practice teach-ing files of eighteen clinical departments, 4 scored more than 90, 8 scored between 80 to 89, 3 scored between 70 to 79, 3 scored between 60 to 69. Three advanced departments are obstetrics and gynecology, auxiliary department and emergency department. There were no significant differences in the scores of major surgery and internal medicine. Compared with that of other departments in the hospital, the specialty of hospital had low scores. It showed that most of the clinical practice of the file information was relatively sound with complete records and some departments had missing items or incomplete and non-standard doc-umentation records. Conclusion Qualitative study on practice teaching files can reflect clinical practice teaching quality. And it is meaningful for us to enhance teaching files construction and to improve clinical practice teaching levels.

Objective To investigate the clinical value of amplitude integrated electroencephalogram on early diagnosis and prognosis evaluation of brain injury caused by neonatal asphyxia.Methods A total of 34 full-term asphyxiated neonates(asphyxia group)hospitalized in NICU of our hospital from January 2015 to September 2015 were selected;meanwhile,34 full-term healthy infants(control group)of the same term were selected.All cases were monitored for the activities of aEEG background,sleep-awakening cycle(SWC)and epileptic activity(SA)within 6 hours after birth.Meanwhile,the relationships between various indexes and asphyxia degree and brain injury were analyzed.Results The electroencephalogram of the asphyxia group was 52.9％and the rate of SWC was 58.8％,which were lower than those of the control group,and the difference had statistic significance(P<0.05).Meanwhile,neonates with epileptic activity in asphyxia group accounted for 11.8％,which was higher than that of control group significantly(P<0.05).Conclusion The AEEG changes of neonates at early period after birth are closely related to perinatal asphyxia and brain injury after asphyxia.The application of amplitude integrated electroencephalogram has an important significance on early diagnosis of neonatal asphyxia.

Objective To investigate the diagnosis and treatment of visceral artery aneurysms (VAAs).Methods From June 2008 to December 2013,20 patients (11 male and 9 female) with visceral artery aneurysms were treated in our hospital.The clinical data and treatment outcomes were retrospectively analyzed.Results There were 20 patients,including 11 males and 9 females.The site of aneurysmal disease was splenic artery in 12 cases,mesenteric artery in 2 cases,hepatic artery in 2 cases,renal artery in 2 cases,celiac artery in 1 case and gastroduodenal artery 1 case.10 patients were treated by open surgery,and other 10 patients treated by endovascular.1 patient died,and others were discharged.All patients were followed up 28.5 months (ranged from 10 to 76 months).Two patients died due to cirrhosis of the liver,and the remaining patients were alive,no treatment-related complications,no serious complications,such as aneurysm recurrence occurs.Conclusions Both traditional surgical and endovascular treatment of visceral artery aneurysms may obtain a satisfactory outcome.

ObjectiveTo investigate the expression of MUC1 in thyroid cancer and benign thyroid diseases (nodular goiter and adenoma) and its clinical significance. Methods The expression level of MUC1 was examined by immunohistochemical analysis with MUC1 monoclonal antibody in 68 samples of thyroid carcinoma and in 18 samples of benign diseases and 10 normal thyroid tissues. Results Immunohistochemical analysis showed positive expression of MUC1 in 51 out of 68 cases of thyroid cancer, in 4 out of 18 cases of benign thyroid diseases and 1 out of 10 normal control thyroid tissue, with a difference statistically significant between the malignancy and benign and normal control (? 2=17.20,P0.05).Conclusion The over-expression of MUC1 in thyroid cancer may act as diagnostic markers of thyroid carcinoma.

Objective To investigate the expression of MUC1 in primary liver carcinoma (PLC) and in cirrhotic liver tissue and its clinical significance in the diagnosis and immuotherapy of PLC. Methods The expression of MUC1 was examined by immunohistochemical analysis in 43 samples of primary liver carcinoma (PLC) , incluing 34 samples of hepatocellular carcinoma (HCC), 9 samples of cholangiocarcinoma (CC) ;and 12 samples of cirrhotic liver tissue and 6 samples of normal liver tissues. Results Immunohistochemical analysis showed over- expression of MUC1 in PLC , which aberrantly localized in the cancer cell membrane; while expression of MUC1 was detected only in 2 cirrhotic liver samples,and no expression in normal liver tissue. The expressional level of MUC1 in PLC was significantly higher than that in cirrhotic and normal liver tissue ( P 0.05),but the expressional levels between with portal cancer emboli group and without portal cancer embloli group was significant difference( P