In order to increase antigenicity of H. pylori-specific antigens and decrease the cost of further industrial production, we used a special PCR with linking primers to construct a fusion gene containing H.pylori ureB and hpaA genes and E.coli ltB gene, and to costract its prokaryotic expression system pET42a-ltB-ureB-hpaA-E.coliBL21DE3. The sequencing result indicated the 100% nucleotide sequence homology of the constructed ltB-ureB-hpaA fusion gene compared to those of the original separated genes. Output of the target recombinant protein rLTB-UreB-HpaA was approximate 15% of the total bacterial proteins measured by SDS-PAGE. The rLTB-UreB-HpaA could induce the immunized rabbits to produce specific antibodies with immunodiffusion titer of 1∶8, and could combine to the commercial rabbit antibody against the whole cell of H.pylori as well as rabbit anti-UreB and anti-HpaA sera by using Western bolt assays. Using GM1-ELISA, the ability of rLTB-UreB-HpaA binding to bovine GM1 was confirmed.And rLTB-UreB-HpaA (200 μg per mouse) could prevent 100% of the immunized BaLb/C mice from H.pylori strain SS1 infection. The co-administration with 10 μg rLTB, the rUreB or rHpa could increase its protective rates in the immunized mice from 66.7% to 81.8% and 83.3%, respectively. All these data leads a conclusion that rLTB-UreB-HpaA is a great potential as a practical genetic engineering vaccine to prevent H.pylori infection.

Objective:To investigate the expression of microRNA (miR)-938, its effect on cell proliferation and its regulatory mechanism in hepatocellular carcinoma (HCC).Methods:HCC and paracancerous tissues were collected from 40 patients with HCC who underwent surgical resection in the Second Affiliated Hospital of Nantong University from January 2015 to June 2019, including 25 males and 15 females, with an average age of 61.4 years. HepG2 cells in the miR-938 overexpression group were transfected with miR-938 mimics, and the negative control group was transfected with the negative control sequence. Cell proliferation was detected by kit, the expression of miR-938 and the succinate dehydrogenase complex subunit D (SDHD) was detected by fluorescence quantitative polymerase chain reaction, and SDHD protein expression was detected by Western blot. The target genes of miR-938 were verified by dual luciferase reporting.Results:The relative expression of miR-938 in HCC tissues was (0.060±0.002), which was higher than that in adjacent tissues (0.030±0.002), and the difference was statistically significant ( P<0.05). The mRNA relative expression of SDHD in HCC tissues was (0.028±0.002), lower than that in adjacent tissues (0.062±0.002), and the protein expression of SDHD in HCC tissues was (0.963±0.008), lower than that in adjacent tissues (1.083±0.037), with statistical significance (both P<0.05). The proliferation activity of miR-938 overexpression group was significantly higher than that of negative control group, and the difference was statistically significant ( P<0.05). MiR-938 significantly inhibited the luciferase activity of SDHD wild-type 3’-untranslated regions. In the overexpression miR-938 cells, SDHD mRNA and protein levels were significantly lower than those in the negative control group ( P<0.05). Conclusion:MiR-938 was highly expressed in HCC tissues. MiR-938 promoted the proliferation of HCC cells by inhibiting the expression of SDHD.

Objective To prepare a gelatin hydrogel crosslinked by genipin and loaded with silver sulfadiazine(AgSD) nanoparticles,and test its in vitro anti-bacterial activities and in vivo wound healing enhancing properties. Methods The suspension of AgSD nanoparticles and coarse powder,hydrogels loaded with AgSD nanoparticles and coarse powder were prepared,respectively, and blank gels were used as control. The diameters of inhibition zone of sensitive bacteria were measured;The biocompatibility to L929 cells was studied by MTT method and relative growth rates of the cells were determined. The minimum inhibitory concentration (MIC)and minimum bactericidal concentration(MBC)of 3 sensitive bacteria were tested;DeepⅡdegree scald co-aureus infection mice model was established and the healing index of treatment by drug-loaded hydrogel dressing was calculated within 3 weeks. The collagen deposition was studied after sirius red staining and pathological manifestations were observed after HE staining. Results Compared with AgSD coarse powder,the antibacterial property of AgSD nanoparticles obviously improved in inhibition zone and MIC, MBC experiments. Better biocompatibility and antibacterial property were revealed for AgSD nanoparticles gel group compared with AgSD cream group in MTT assay and inhibition zone studies;Healing index increased significantly since the second week in AgSD nanoparticles gel group compared with staph group,AgSD cream group and AgSD coarse powder gel group in wound-healing experi-ment(P<0.05). Compared with those control groups,nanocrystal gel group had more content of typeⅠcollagen and total collagen, and higher pathology scores(P<0.05). Conclusion AgSD nanocrystal loaded gelatin hydrogel has good antibacterial activity in vitro and high quality healing enhancing property in vivo.

Objective To prepare a silver sulfadiazine(AgSD)-nanocrystal loaded hydrogel with gelatin as the main raw mate-rial,genipin as crosslinker,and test its physicochemical property and drug release characteristics. Methods AgSD-nanocrystal was prepared by ball milling,the particle size and polydispersity index were determined,the solubility was studied by an ultra perfor-mance liquid chromatography(UPLC)method and the microstructure was observed with transmission electron microscope(TEM);drug loaded gelatin hydrogel in different crosslinking degrees were prepared,the dissolution rate was studied in the same way as AgSD-nanocrystal and the possible dissolution mechanism was analyzed by fitting curves;the microstructure was observed by scanning elec-tron microscope(SEM). After 6 weeks,with the same method,all the above mentioned properties of drug loaded hydrogel and AgSD-nanocrystal suspension were determined repeatedly,and at the same time their stability was tested. Results After nanocrystalliza-tion,the polydispersity index of AgSD was 0.211,the mean particle size was about 267.8 nm,the dissolution rate greatly increased within 6 hours compared with that of AgSD bulk powder. There was a negative correlation between dissolution rate and crosslinking de-gree,and the nanoparticle gel's curve was higher than that of the bulk. In terms of stability,AgSD-nanocrystal in hydrogel was almost in the original state in SEM,while those in suspention had an increasing particle size and decreasing dissolution rate. Conclusion The stability and dissolution of the new drug loaded hydrogel are good,and its data can be used as reference for relevant study.

Objective: To explore influence of exercise on cardiac autonomic nerve function in patients undergoing coronary artery bypass grafting (CABG) and its clinical significance. Methods: A total of 53 patients with coronary heart disease undergoing CABG were randomly divided into routine treatment group (n=25) and rehabilitation group (n=28, received exercise training based on routine treatment). Changes of autonomic nerve function before, second and eighth week after CABG were analyzed in two groups using time domain indexes of heart rate variability (HRV) by ambulatory electrocardiography, including standard deviation of normal to normal RR intervals calculated over the 24 h period (SDNN), standard deviation of normal to normal RR intervals in all 5 min segments of the entire recording (SDANN), root-mean square of differences between successive normal to normal intervals (rMSSD) and adjacent normal RR interval difference > 50ms stroke accounted for a percentage of 24h total RR interval (PNN50). Results: Compared with before CABG, there were significant decrease in SDNN, SDANN, RMSSD and PNN50 in both groups two weeks after CABG (P<0.05~0.01); eight weeks after CABG, above indexes recovered to levels before CABG in routine treatment group[SDNN （113.6±29.4）vs.（116.7±24.7）, SDANN（112.2±32.1）vs.（113.6±28.6）, rMSSD（21.9±8.2） vs.（23.2±7.1）, and PNN50 （7.5±4.2）vs.（8.2±3.7）] , P>0.05 all; Compared with before CABG, there were significant improvements in SDNN [(114.7±25.2) ms vs. (132.6±30.6) ms], SDANN [(111.6±23.5) ms vs. (129.2±30.8) ms], rMSSD [(24.2±8.7) ms vs. (29.9±7.5) ms] and PNN50 [(7.8±2.2) ms vs. (9.5±2.3) ms], and there were significant improvement than those of routine treatment group [SDNN （132.6±30.6）vs.（113.6±29.4）, SDANN（129.2±30.8）vs.（112.2±32.1）, rMSSD（29.9±7.5）vs.（21.9±8.2）and PNN50 （9.5±2.3）vs.（7.5±4.2）] eight weeks after CABG in rehabilitation group, P<0.05 all. Conclusion: All HRV indexes significantly decrease on two weeks after CABG in both groups, suggesting that CABG can damage cardiac autonomic nerve system. These indexes of rehabilitation group were more improvement than those of routine treatment group, suggesting exercise training can more significantly improve cardiac autonomic nerve function after CABG.

0.35 U?L-1 was taken as standard of positive detection.Among all the 20 allergen,atopy could be diagnosed by one positive allergen detected.The controlling non-atopy group were the controlls.Reverse transcription polymerase chain reaction assay was used to detect viruses in the nasopharyngeal secretions of these patients,including respiratory syncytial viruses,rhincvirus,influenza virus,parainfluenza,human metocpneumo virus,human bocavirus,enterovirus.The virus-positive patients were then divided into 2 groups,atopic virus-positive group and non-atopic virus-positive group.Cytokines IL-12 and IL-27 were further determined using enzyme-linked immunosorbent assay me-thod.Results A total of 65 cases(56.0%) and 77 cases(66.4%) out of 116 cases of recurrent wheezing children,were found to be allergen-positive and virus-positive,respectively.The virus-positive rate was 75.4% in atopic group and 54.9% in non-atopic group.There was a significant difference in the virus-positive rates between the atopic and non-atopic group(?2=5.37,P0.05).Furthermore,serum IL-12 and IL-27 in the atopic group were significantly higher than those in the non-atopic group(t=2.579,2.573,Pa

Objective To investigate the correlation of position movement of primary tumor with interested organs and skin markers,and to investigate the correlation of volume variation of primary tumors and lungs during different respiration phases for patients with lung cancer at free breath condition scanned by four-dimensional CT (4DCT) simulation.Methods 16 patients with lung cancer were scanned at free breath condition by simulation 4DCT which connected to a respiration-monitoring system.A coordinate system was created based on image of T5 phase,gross tumor volume (GTV) and normal tissue structures of 10 phases were contoured.The three dimensional position variation of them were measured and their correlation were analyzed,and the same for the volume variation of GTV and lungs of 10 respiratory phases.Results Movement range of lung cancer in different lobe differed extinct:0.8 - 5.0 mm in upper lobe,5.7 -5.9 mm in middle lobe and 10.2 - 13.7 mm in lower lobe,respectively.Movement range of lung cancer in three dimensional direction was different:z-axis 4.3 mm ± 4.3 mm＞ y-axis 2.2 mm ± 1.0 mm ＞ x-axis 1.7 mm ± 1.5 mm ( x2 =16.22,P =0.000),respectively.There was no statistical significant correlation for movement vector of GTV and interested structures (r =-0.50 - -0.01,P =0.058 - -0.961 ),nor for volume variation of tumor and lung ( r =0.23,P =0.520 ).Conclusions Based on 4DCT,statistically significant differences of GTV centroid movement are observed at different pulmonary lobes and in three dimensional directions.So individual 4DCT measurement is necessary for definition of internal target volume margin for lung cancer.

Objective To investigate the sedative and hypnotic interaction between remifentanil and propofol by target-controlled infusion (TCI) during induction of anesthesia.Methods Third-two ASA Ⅰ or Ⅱpatients,aged 22-63 yr,body mass index 18-25 kg/m2,scheduled for elective surgery under general anesthesia,were randomly divided into 4 groups(n=8 each).Group Ⅰ only received TCI pmpofol.GroupⅡ,Ⅲ,and Ⅳreceived a target concentration of 2,4 or 6 ng/ml remifentanil respectively.While the blood-effect site concentrations of remifentanil were equilibrated,patients received TCI of propefol,with an initial target concentration of 0.5μg/ml.After the blood-effect site concentrations of propofol were equilibrated then with 0.5μg/ml increments until the loss consciousness was achieved.The eyelash reflex and state of consciousness were assessed and radial arterial blood sample 6 ml was taken every 3 min to determine the remifentanil and propofol concentrations in blood.Propofol and remifentanil concentrations in blood were measured by reversed-phase high-performance liquid chromatography and high-performance liquid chromatography with ultraviolet detection respectively.The sedative and hypnotic interaction between propofol and remifentanil was determined with a pharmacodynamie interaction model by regression analysis and determined using the isobolographic method.Results Propofol concentrations in blood were lower in group Ⅱ,Ⅲ and Ⅳ than group Ⅰ(P<0.05).The propofol concentratopms in blood were significantly decreased in trun with the increase in the remifentanil concentrations in blood in group Ⅱ-Ⅳ(P<0.05).At loss of eyelash reflex and loss of consciousness of patients,the pharmacodynamic interaction model by curve fitting was superior to linear regression (P<0.05).At loss of eyelash reflex of patients,the curve fitting result showed EC50,prop=2.77μg/ml and EC50,rem=26.67 ng/ml,and the isobolographic method equation is ECprop/2.77+ECrem/26.67=0.69.At loss of consciousness of patients,the curve fitting result showed EC50,prop==3.76μg/ml and EC50,rem=31.56ng/ml,and the isobolographic method equation is Ecprop/3.76+Ecrem/31.56=0.65.Conclusion Remifentanil (Cp 2-6 ng/ml) and propofol by TCI shows a synergistic type of pharmacodynamic interaction on the sedative and hypnotic during induction of anesthesia.

BACKGROUND: Biological characteristics of porcine cardiovascular system are similar to that of human. The metabolism, immune system, mechanism of disease have 99% homology between pig and human. OBJECTIVE: To develop ventricular aneurysm animal models by plugging left anterior descending branch. DESIGN, TIME AND SETTING: The animal observational study was performed at the Laboratory of Functional Material and Animal Experimental Center, Tongji Hospital Affiliated to Tongji University from December 2005 to July 2007. MATERIALS: A total of 13 pigs, of both genders, weighing 30-40 kg, were used in this study. METHODS: After abdominal cavity and intravenous anesthesia, balloon with "WYW" stent was put into 13 pig left anterior descending coronary artery following No. 7 artery sheathing canal was implanted. The balloon was dilated and the stent was positioned into the distal point of the first diagonal branch to obstruct the artery under digital subtraction angiography. During and after the operation, electrocardiogram was monitored and recorded to maintain stable vital sign. MAIN OUTCOME MEASURES: Electrocardiogram, serum myocardial enzymes, myocardial radionuciide imaging, echocardiography and angiography and pathological changes were observed. RESULTS: One rat died of anesthetic accident before surgery. Six rats died from ventricular fibrillation during plugging, and the other 6 rats were survival. 4 weeks following surgery, coronary angiography showed blood flow at distal end was blocked 100%. Left ventricle angiography demonstrated that wall motion at apex of the heart and the left ventricle disappeared. Before embolism, electrocardio-monitoring displayed electrocardiogram was normal. After plugging, ST segment was raised continuously; amplitude of R wave was decreased; T wave was erected; ST-T fusion wave appeared; precordial leads were obvious. ST segment was reduced to basic level 2 week later, and pathological Q wave appeared 4 weeks later. Serum treponin was increased at 12 hears following surgery compared with before surgery (P < 0.01). Radioactive nuclide myocardial imaging revealed that cardiac apex and left ventdcle anterior wall showed radioactive nuclide perfusion filling defect, thin ventricular wall and ventricular aneurysm. Cardiac ultrasonic showed six pigs suffered from weaken segmentation contraction motor of local ventricular wall, especially at anterior wall of the left ventricle and cardiac apex. After hematoxylin-eesin staining, optical microscope exhibited that cardiac muscle fiber disappeared, which was replaced by collagen fiber, with the presence of some capillary at the infarct of cardiac apex. Pyknosis and lysis of residual cardiac muscle fiber were visible in infarct region of anterior wall of the right ventricle. Many inflammatory cells and capillary infiltration were detected besides collagen fiber filling. CONCLUSION: The new approach of "WYW" intervention embolization is able to develop the ventricular aneurysm animal model, which is similar to clinical pathophysiological variation. The outcome is precise and reliable.

Compared with living spray method, it focused on the investigation of different inoculation methods, various inoculation concentration and the influence of different seeding age on soft rot-resistance in Jinxianlian. The results showed that (1) Inoculated with dropping connection, the difference of disease index between A. roxburghii and A. formosanus was grate, so that the disease-resistance could be obviously distinguished. (2) When the inoculation concentration was 1.0 x 10(7) cfu x mL(-1), the difference of disease index was relatively obvious and the disease-resistance could be differentiated well. (3) At the moment of 4-month seeding inoculation, a certain difference of the disease index between A. roxburghii and A. formosanus was existed, so, relatively, it could accurately reflect the resistance difference between various species. With the inoculation of dropping connection, A. roxburghii and A. formosanus of 4-month seeding age was put in the bacteria suspension of inoculation concentration of 1.0 x 10(7) cfu x mL(-1). The identification was taken up after 5 days in the incubator under the condition of 14 h daylight and 28 degrees C. The identification result was conformed with that of the living spray method. To investigate the identification method of in vitro evaluation of soft rot-resistance of Jinxianlian so as to provide the foundation for germplasm utilization and excellent cultivars breeding.
Plant Diseases ; microbiology