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: To explore the effect of moxibustion at Shenque (CV 8) on myocardial structure and function in exercise-induced fatigue rats. Methods: A 12-week treadmill running training was performed to create an exercise-induced fatigue rat model. Sixty eligible male specific-pathogen-free grade Sprague-Dawley rats were randomly divided into a blank group, a control group, a model group, a non-meridian non-acupoint group, a Zusanli (ST 36) group and a Shenque (CV 8) group, with 10 rats in each group. Rats in the blank group did not receive treadmill running training or moxibustion. Rats in the control group did not receive treadmill running training but received mild moxibustion at Shenque (CV 8). Rats in the model group received treadmill running training but no moxibustion. Rats in the non-meridian non-acupoint group, the Zusanli (ST 36) group and the Shenque (CV 8) group received moxibustion at the non-meridian non-acupoint points, Zusanli (ST 36) or Shenque (CV 8) immediately after each treadmill running training, 15 min each time, once a day for 5 consecutive days a week at a 2-day interval, 60 times of moxibustion in total. Left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVESd), left ventricular diastolic volume (LVDv), left ventricular systolic volume (LVSv), ejection fraction (EF), stroke volume (SV), early diastolic peak flow velocity of mitral valve (E) and late diastolic peak flow velocity of mitral valve (A) of each group before and after the last treadmill running training were measured. Blood was collected 6 h after the last treadmill running training, and serum C-reactive protein (CRP), myoglobin (Mb), creatine kinase-myocardial band (CK-MB), cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels were detected. Finally, the heart was separated, the heart mass (HM) was measured, the cTnT level of the myocardial tissue was detected, the ultrastructural changes of the left ventricular myocardium were observed by transmission electron microscope, the left ventricular fraction shortening (LVFS), E/A and heart mass index (HMI) were calculated. Results: Compared with the same group before treatment, the rat cardiac LVEDd, LVESd, LVDv, LVSv, SV, E and A were significantly increased (all P<0.01), and the rat LVFS, E/A and EF were significantly decreased (all P<0.01) in the model group and the non-meridian non-acupoint group after treatment; the rat cardiac SV, LVDv, LVSv, E and A were all increased (all P<0.01), while E/A and EF were decreased (all P<0.01) in the Zusanli (ST 36) group after treatment; the rat cardiac LVDv, E and A were significantly increased (P<0.01 or P<0.05), and E/A was significantly decreased (P<0.01) in the Shenque (CV 8) group after treatment. After treatment, compared with the blank group, the rat cardiac LVEDd, LVESd, SV, LVDv, LVSv, E, A, the serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and the myocardial cTnT were increased (all P<0.01), and the LVFS, E/A and EF were all reduced (all P<0.01) in the model group; compared with the model group and the non-meridian non-acupoint group, rats in the Zusanli (ST 36) group and the Shenque (CV 8) group showed decreased LVEDd, LVESd, SV, LVDv, LVSv, E, A, serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and myocardial cTnT (P<0.01 or P<0.05), along with increased LVFS, E/A and EF (all P<0.01); compared with the Zusanli (ST 36) group, Mb and A of the Shenque (CV 8) group were decreased (both P<0.01), while both E/A and EF were increased (P<0.01, P<0.05). Transmission electron microscopy examination showed that myofibrils in the blank group and the control group were neatly arranged with clear light and dark bands; the model group and the non-meridian non-acupoint group showed different degrees of myofibril disintegration and breakage, increased and aggregated mitochondria of different sizes, and increased electron density. The myofibrils in the Shenque (CV 8) group and Zusanli (ST 36) group were arranged neatly with clear light and dark bands, and compensatory hyperplasia of mitochondria. Conclusion: Moxibustion at Shenque (CV 8) and Zusanli (ST 36) both can effectively improve the occurrence of myocardial remodeling in exercise-induced fatigue rats, and the effect of moxibustion at Shenque (CV 8) is better in improving cardiac function.

OBJECTIVETo study the epidemiological characters and risk factors of renal calculi in Shenzhen for future intervention.

METHODSRisk factors of renal calculi were analyzed by factor analysis and linear structural relation model, followed by fitting and evaluating the model.

RESULTSThe prevalence rates of renal calculi were 8.00% and 5.12% in males and females. The results of LISEREL showed that the total effect of age and sex affecting the development of renal calculi was 0.4614, with directly effect 0.3600 and indirect 0.1014. The direct effects of blood uric acid, blood calcium and body mass index, blood cholesterol, blood sugar were 0.3400 and 0.3200 respectively. The indirect effect of education, exercise and dietary habit affected the development of renal calculi through index of biochemistry and obesity, were -0.0416 and 0.1882.

CONCLUSIONSex, age, obesity and high blood cholesterol, high blood sugar, high blood uric acid, high blood calcium were the direct influencing factors to renal calculi. At the same time, education, exercise and dietary habit were also associated with the disease.

Age Factors ; Blood Glucose ; metabolism ; Body Constitution ; physiology ; Calcium ; blood ; China ; epidemiology ; Cholesterol ; blood ; Exercise ; physiology ; Feeding Behavior ; physiology ; Female ; Humans ; Kidney Calculi ; epidemiology ; Linear Models ; Male ; Prevalence ; Risk Factors ; Sex Factors ; Social Class ; Socioeconomic Factors ; Uric Acid ; blood

Objective To discuss the views and significance of the "parallel" training mode, and provide a reference for the smooth development of the graduate students' standardized training work. Methods The self-designed questionnaire included rotation/training, clinical skills, scientific research, supporting conditions, and the rationality and prospect of the integrated training mode. We chose the graduate students of professional type classes of Grade 2014 and 2015 in a university affiliated hospital of Kunming, in a secret-based manner , and questionnaires were distributed and collected by stratified sampling and random sampling according to the proportion of 50％ of the total number of them. Excel database was used for data processing and analysis. Results A total of 200 questionnaires were distributed and 167 were recovered with an effective recovery rate of 83.5％. The research items involved in the investigation of the students that were more satisfied with the project were as follows:methods of integrated training mode being more reasonable (85.0％, n=142), improving clinical comprehensive ability (83.2％, n=139), accelerating the development of high-level clinical talents (67.7％, n=113), conducive to unify the clinical practice standards (55.1％, n=92), conducive to employment (48.5％, n=81), saving medical resources (44.3％, n=78). Meanwhile, the survey found that graduate students were not satisfied with the following items: research time (88.0％, n=147), arrangements for plans of rotate conversion (65.8％, n=110), monthly income (89.8％, n=150). Conclusion The integrated training mode can accelerate the development of high-level clinical talents, save medical resources, further improve the medical education system and better improve the clinical com-prehensive ability of medical students. However, it still needs to be further improved in terms of the arrange-ments for plans of rotate conversion of clinical departments, cultivation of scientific research ability, and salary, etc.

After the characteristics of project-driven teaching methods and traditional teaching methods were com-paratively analyzed,a new gradient and hierarchical "dual-driven" model of medical interdisciplinary innovative informationists training was established by embedding the general model of project-driven teaching methods into the traditional training model of medical interdisciplinary innovative informationists training in order to improve the teaching level and find a new way for medical informationists training.

OBJECTIVETo study the use of neural network in determining the risk factors of diseases.

METHODSWith back-propagation neural network (BP network) as fitting model based upon data gathered from an epidemiological survey on diabetes mellitus and under the network structure of 22-6-1, the mean impact value (MIV) for each input variables and sequencing the factors according to their absolute MIVs were calculated. The results from BP network with multiple logistic regression analysis and log-linear model for united actions between factors were compared with optimizing Levenberg-Marquardt algorithm.

RESULTSBy BP network analysis, the sequence of importance for the risk factors of diabetes mellitus became: faster pulse, diabetes mellitus family history, living longer in the investigated area, with medical record of nephropathy, having higher ratio for waist-to-hip, being male, with medical records of diseases as hyperlipoproteinmia, coronary heart disease, hypertension, high diastolic pressure, higher income, do no drink alcohol, age, higher systolic pressure, less educated, body mass index, with medical records of other diseases, physical exercise related to jobs smoking, occupation, with medical record for cerebrovascular disease, with medical record for liver disease etc. However, only 7 factors were statistically significant in multiple logistic regression analysis. The sequence of their importance appeared as: pulse, diabetes mellitus family history, the medical record of nephropathy, waist-to-hip ratio, the medical record of hypertension, work-place related exercise and age. The sequences of importance were almost the same between the two while the difference could partly be explained by the interaction among risk factors through log-linear model.

CONCLUSIONNeural network could be used to analyze the risk factors of diseases and could assimilate more complicated relationships (main effects and interactions) between inputs and outputs, better than using the traditional methods.

Adult ; China ; epidemiology ; Diabetes Mellitus ; epidemiology ; etiology ; Family Health ; Female ; Humans ; Hyperlipidemias ; complications ; Logistic Models ; Male ; Neural Networks (Computer) ; Obesity ; complications ; Pulse ; Risk Factors

OBJECTIVETo examine the significance of individual risk on diabetes to subjects who underwent diabetes screening.

METHODS2003 asymptomatic diabetes subjects with high-risk factors of diabetes as family history, obesity, hypertension, and/or dyslipidemia, fetal giant history were screened. 5362 subjects having no risk factors but from the same community were allocated as controls.

RESULTSThere were 131 (6.54%) diabetes identified in the screening group and 1547 (77.23%) subjects having 1 risk factor, 387 (19.27%) having 2 risk factors, 70 (3.49%) having 3 or more risk factors. There were 96 (1.79%) diabetes identified in the control group. Compared with control group, the OR (95% CI) value was 2.68 (2.20-3.25) after adjusted on age among the high risk group. The OR value of those having 1 risk factor was 2.89, but these having 3 or more risk factors increased to 4.68.

CONCLUSIONThe relation between the risk of high-risk group with diabetes and the number of risk factors of diabetes presented positive correlation. Early and regular screening for diabetes was essential in these individuals with high-risk factors.

Adult ; Aged ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; etiology ; genetics ; Family Health ; Female ; Glucose Tolerance Test ; Humans ; Hyperlipidemias ; epidemiology ; Hypertension ; epidemiology ; Male ; Mass Screening ; Middle Aged ; Obesity ; epidemiology ; Odds Ratio ; Prevalence ; Risk Factors ; Smoking

OBJECTIVETo evaluate the prognostic significance of p53 mutation and P53 protein expression abnormality among esophageal cancer.

METHODSThe results of 27 random controlled trials from 1990 to 2003 were analyzed by meta-analysis method. The overall positive rate of p53 was 52.9% among the cumulative 2174 cases. Relative hazard (RH) was applied to evaluate the risk of disease and all data were analyzed by Dersimonian-Laird method.

RESULTSThe analysis for homogeneity (q statistics test) showed that all eligible studies were with heterogeneity (q = 59.88, P < 0.005). The combined RH was 2.07 and 95% confidence interval was 1.58-2.70.

CONCLUSIONFindings showed that p53 was a poor prognosis biomarker for esophageal cancer gene diagnosis but might benefit to the strategy of treatment.

Carcinoma, Squamous Cell ; genetics ; metabolism ; Esophageal Neoplasms ; genetics ; metabolism ; Female ; Genes, p53 ; genetics ; Humans ; Male ; Mutation ; Prognosis ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVESTo investigate the expression of cyclooxygenase-2 (cox-2) protein in normal squamous epithelium, squamous dysplasia and squamous cell carcinoma of the esophagus, and to elucidate the role of cox-2 in esophageal carcinogenesis.

METHODSBiopsy specimens of atypical esophageal dysplasia (n = 47) and surgical resection of squamous cell carcinoma (n = 86) were compared with normal esophageal specimens (n = 42) and the expression of cox-2 in those specimens was analyzed by immunohistochemistry and western blotting.

RESULTSA significant elevated cox-2 expression was shown in atypical esophageal squamous dysplasia and squamous cell carcinoma, as compared to that in normal esophageal squamous epithelium, with immunohistochemical stain scores of 2.67 +/- 1.77, 2.19 +/- 1.79 and 0.71 +/- 0.46, respectively. Results of western blotting analysis confirmed those obtained by immunohistochemistry. Cox-2 expression significantly correlated with proliferation activity assessed by proliferating cell nuclear antigen index in dysplastic and carcinomous lesions, respectively, and no such correlation could be found in normal esophageal mucosa. Elevated cox-2 expression was not associated with clinical-pathological features of esophageal squamous carcinoma, including age, gender, tumor size, histological grade, lymph node metastasis and clinical stage.

CONCLUSIONElevated expression of cox-2 in atypical squamous dysplasia and squamous cell carcinoma of the esophagus, which correlated with cell proliferation activity, indicated that cox-2 may be involved in the early stage of squamous carcinogenesis of the esophagus, and may be a target of prevention and treatment for esophageal squamous cell carcinoma.

Adult ; Aged ; Blotting, Western ; Cyclooxygenase 2 ; Esophageal Neoplasms ; enzymology ; pathology ; Female ; Humans ; Immunohistochemistry ; Isoenzymes ; biosynthesis ; Male ; Membrane Proteins ; Middle Aged ; Neoplasms, Squamous Cell ; enzymology ; pathology ; Prostaglandin-Endoperoxide Synthases ; biosynthesis

ObjectiveTo describe the modified (narrowed) gastric tube and two methods on the establishmentof enteral feeding pathway in esophagectomy.MethodsFrom July 2003 to April 2006,we made traditional gastric tube for patients underwent esophagectomy to substitute the dissected esophagus while from May 2006 to November 2009,we made modified gastric tube for the same purpose.From July 2003 to November 2009,enteral nutrition tubes for all these patients are placed intraoperatively by two different ways,which made early post-operative nutrition support possible for these patients.ResultsNo perioperative death occurred among the patients.The length of the modified gastric tube ensured the anastomosis by circular stapler at the apex thorax or in the bases of cervical region.No statistical differences were found between the two group in terms of survival rate.The intraoperative establishment of enteral nutrition pathway ensured the early enteral nutrition support after the operation.ConclusionThe utility of the modified gastric tube extends the length of gastric tube to make mechanical anastomose easier and safer; meanwhile,the intraoperative establishment of enteral nutrition pathway ensures the early enteral nutrition support after the operation.