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

A total of 60 ankylosing spondylitis patients scheduled for elective surgery with anticipated difficult airway were enrolled and randomly assigned into either dexmedetomidine (D) or midazolam plus sufentanil (MF) group.Group D:topical nasal anesthesia and a loading dose of dexmedetomidine at 1.0 μg/kg in 10 min; group MF:intravenous infusion of 0.05 mg/kg midazolam plus 3 μg/kg sufentanil.Fiberoptic bronchoscopic (FOB)-assisted awake nasotracheal intubation was performed.Mean arterial pressure (MAP),heart rate (HR),Pulse oxygen saturation (SpO2),Ramsay score and success rate of intubation were recorded and compared between two groups.The intubation conditions and level of comfort were also evaluated.MAP and HR of group D at FOB through postnaris (T1),peep the epiglottis(T2),intubation success immediately(T3)and 1 min after intubation(T4) were significantly lower (P ＜ 0.05 or P ＜ 0.01) than those of group MF while Ramsay sedation scores were obviously higher (P ＜ 0.05) than those of group MF.Group D with endotracheal intubation comfort level 5 score of grade 1-2 were 8 and 14 cases and were significantly higher than 4 and 10 cases of group MF.Immediately after intubation,level 3 scores in grade 1 of group D (n =21) were significantly higher than group MF (n =12) ; initial success rate of intubation in group D was obviously higher than that in group MF (70％,n =21 vs.47％,n =14).And the incidence of unpleasant intubation memory in group D was lower than that in group MF (37％,n =11 vs.67％,n =20).Fiberoptic bronchoscopic-assisted nasotracheal intubation offers better conditions for intubation and reduces the incidence rate of intraoperative awareness.

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

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

This article discussed on the continuing medical education for general practitioners within a nonprofit multihospital system.Literature review,statistical analysis and survey results were employed.Training situation for general practitioner at home and aboard was reviewed.Statistics on population,disease spectrum,present situation of community doctors in Luwan branch of Ruijin hospital was also analyzed.Results showed that current education system for general practitioners cannot catch up the increasing demand yet.Meanwhile patients expected well qualified and skillful physicians in the community healthcare centers.Reinforcement of continuing medical education for general practitioners was the stepping stone of a nonprofit multihospital system.Improving skills and qualifications of physician played an essential role in the urban network of medical services.