Objective To evaluate the MUC1 promoter's role in driving gene expression in pancreatic cancer and its therapeutic significance.Methods Two plasmids were made.The plasmid pEGFP-MUC1N1 contained MUC1 promoter fragment connected to the pEGFP-N1 vector with the EGFG reporter gene.The pShuttle-MUC1-EGFP plasmid contained MUC1 promoter fragment and EGFP reporter gene connected to pShuttle plasmid.Lipofectamine 2000 was used to transfect the two plasmids into cells of MUC1-positive human pancreatic cell line Panc-1 and MUC1-negative human cervical carcinoma Hela.Fluorescence microscopy and flow cytometry compared the specificity and activity of the MUC1 promoter and CMV promoter.Results Reporter gene EGFP-positive cells 48 hours after transfection with pEGFP-MUC1-N1 and pShuttleMUC1-EGFP plasmid were 69.6％ and 63.6％ respectively,in Panc-1 cells,and 4.2％ and 3.7％ respectively,in Hela cells.Conclusions MUC1 promoter can drive reporter gene activity in MUC1-positive tumor cells targeting functional expression.There is potentially a use of targeted therapy in pancreatic cancer at the genetic level.

OBJECTIVETo analyze the electrical activity property changes in nucleus accumbens (NAc) of heroin-induced conditioned place preference (CPP) rats during different stages of heroin dependence and to explore NAc's roles in the formation of drug dependence.

METHODSRecording electrodes were bilaterally embedded into the NAcs of rats with the aid of stereotaxic apparatus, followed by establishment of heroin-dependent rat model. The NAc electrical activity during 3 different stages of heroin dependence, including heroin pre-exposure, immediate post-exposure and heroin withdrawal, were respectively recorded using EEG wireless telemetry techniques. The frequency distribution (ranging from 0.5 to 30 Hz) and the amplitude of NAc electrical activity were analyzed and measured.

RESULTSHeroin-dependent rat models were successfully established and their withdrawal symptoms were evident. All rats showed a conditioned place preference (CPP) for the white box after 5-10 days of heroin-exposure, and displayed a maximum withdrawal symptoms on 2d after heroin- withdrawal. During all statges of heroin-dependence, the NAc electrical activity contained the highest proportion of delta rhythm and the lowest proportion of alpha2 rhythm. The discharge frequence band was similar across different stages. There was a significantly increased ratio of low-frequency discharges (delta rhythm) and decreased ratio of high-frequency discharges (beta rhythm) in NAc of rats during the immediate post- heroin exposure stage when compared with that during pre-exposure and heroin withdrawal stages. During the withdrawal stage, the ratio of at rhythm was significantly lower than during pre- and post-heroin exposure stages (P < 0.01). Further, the mean discharge amplitude in NAcs during immediate post-exposure and withdrawal stages was significantly increased relative to pre-exposure stage. However, the mean discharge amplitude during heroin withdrawal stage was significantly lower than during immediate post-exposure stage.

CONCLUSIONThe electrical activity properties in rat NAcs showed a significant change during different stages of heroin-dependence, which suggested that neuronal activities in NAcs might contribute to the modulation of drug-dependence.

Animals ; Conditioning, Operant ; Heroin ; pharmacology ; Heroin Dependence ; physiopathology ; Male ; Nucleus Accumbens ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Telemetry

ObjectiveTo explore the methods of low temperature preservation for alginate-polylysine-alginate (APA) microcapsules.MethodsAPA microcapsules were prepared with static electricity, and underwent hypothermal treatment respectively through methods of program control, gradient by icebox and put in liquid nitrogen directly, finally preserved in liquid nitrogen. The form and permeability of APA microcapsules were checked after rewarming.ResultsThe rates of integrity, crenation and damage were (91.2±1.57)%, (3.1±0.81)% and (5.7± 2.62 )% in the program control group; (85.3±1.42)%, (5.2±0.74)% and (9.5± 3.81 )% in the gradient by icebox group; (14.5±1.57)%, (84.1±3.47)% and (1.4±2.62)% in the directly put in liquid nitrogen group. The membrane permeability of full APA microcapsules after frozen and reworming was not changed obviously.ConclusionThe program control method can preferably preserve APA microcapsules at low temperature and keep them having normal form and permeability.

BACKGROUND: Genetically engineered cells have been used in the replacement therapy and gene therapy. However, how to select proper donor cells, target cells, and corresponding viral vectors is the most difficult in the therapy.OBJECTIVE: To compare the transduction efficiency of recombinant adenovirus Ad5 and AdSF35, adeno-associated virus rAAV1/2 and rAAV2, and lentivirus LV in bone marrow mesenchymal stem cells (BMSCs) and exogenous gene expression level so as to select the vectors, which can efficiently transduce BMSCs.DESIGN, TIME AND SETTING: Gene engineering controlled observation, performed in the Central Laboratory, Shanghai First People's Hospital between October 2006 and March 2007.MATERIALS: Ten Sprague Dawley rats of clean grade were used to prepare BMSCs. All recombinant viral vectors carded enhanced green fluorescent protein (EGFP) report gene. Ad5 was prepared by the Central Laboratory, Shanghai First People's Hospital. Ad5F35 was gifted by professor Qian Qi-jun from the Second Military Medical University of Chinese PLA. rAAV2 and rAAVI/2 were the products of Benyuan Zhengyang Gene Technique Co.,Ltd. LV was gifted by professor Cuo Li-be from Shanghai Institute of Biochemistry and Cytobiology, Chinese Academy of Sciences.METHODS: Rat BMSCs were in vitro isolated and cultured by density gradient centrifugation. BMSCs of passage 4 were inoculated into 24-well plate at lxlO5/well. One day later, ceils adhered to the wall and allocated to 5 groups. Ad5-EGFP [10, 100,1 000 multiplicity of infection (MOI)], Ad5F35-EGFP (10,10, 1 000 MOI), rAAVI/2-EGFP (1×104,1x10× vg), rAAV2-EGFP(1×104, 1x105vg), and LV-EGFP (30 TU) were respectively added into the 5 groups. BMSCs were transduced for 2 days with Ad virus and separately for 6 days with rAAV and LV virus.MAIN OUTCOME MEASURES: EGFP-positive expression and fluorescence intensity.RESULTS: After twenty-four hours of Ad5-EGFP transduction, EGFP-positive cells were visible under the microscope. With virus dose increasing, EGFP-positive cells increased and fluorescence intensity strengthened. Twelve days later, EGFP-positive cells gradually reduced and fluorescence intensity weakened. For Ad5F35-EGFP, its transduction was basically similar to Ad5-EGFP, but EGFP-positive cell number and fluorescence intensity were increased. After 6 days of rAAV1/2-EGFP or rAAV2-EGFP transduction, EGFP-posidve cell number and fluorescence intensity were decreased. For LV-EGFP transduction, a small amount of EGFP-positive cells could be visible on the second day, and then EGFP-positive ceils and fluorescence intensity were gradually increased until the sixth day.CONCLUSION: Ad5, Ad5F35 and LV could effectively transduce BMSCs cultured in vitro and express exogenous gene.Furthermore, transduction efficiency was correlated with virus dose in dose-dependent manner, rAAVI/2 and rAAV2 had poor/in vitro transduction efficiency.

Adult ; Biopsy ; Fatty Liver ; etiology ; pathology ; Female ; Humans ; Lupus Erythematosus, Systemic ; complications

Objective To systematically and quantitatively evaluate the relationship of rs12252 polymorphism in IFITM3 gene and susceptibility to influenza in Chinese population. Methods The databases of Medline, PubMed, Embase, Web of Science, China National knowledge infrastructure (CNKI), Wanfang and VIP were searched to retrieve the articles which focused on the topic of relationship between the rs12252 polymorphism and susceptibility to influenza in Chinese population. Meta-analysis method was used to comprehensively and quantitatively analyze the enrolled articles. Results Seven papers with a total of 919 influenza cases were involved in this article. Influenza virus involved influenza A (H7N9, H1N1 pmd09, H3N2) and influenza B. Study-based sequential meta-analysis of the study showed that the total sample size of this study have achieved the required sample size to obtain stable positive results. The results of this meta-analysis showed that the rs12252 polymorphism was associated with susceptibility to influenza in Chinese population and that the rs12252C polymorphism in IFITM3 gene was more susceptible to have influenza in Chinese people (C vs T: OR=1.67, 95% CI:1.45-1.92; CC vs TT: OR=2.61, 95% CI:1.97-3.46; TC vs TT: OR= 1.55, 95% CI:1.20-2.00; CC vs TC+TT: OR= 2.01, 95% CI:1.49-2.72; CC+TC vs TT: OR =8.90, 95% CI:4.94-16.06). Conclusion the rs12252 polymorphism of IFITM3 gene was a risk factor for influenza in Chinese population.

Twenty-four soil samples of eight ecosystem-types around the Yellow River source area were investigated for the number and specific composition of soil dematiaceous hyphomycetes by dilution plate technique. And then the co-relationship between genus species of soil dematiaceous hyphomycetes and ecosystem-types was analyzed. The results show that the amount and species distribution of soil dematiaceous hyphomycetes had an obvious variability in different ecosystem-types, and that the dominant genus species varied in the eight ecosystem-types studied, with Cladosporium being the dominant genus in seven of the eight ecosystem-types except wetland. The index of species diversity varied in different ecosystem-types. The niche breadth analysis showed that Cladosporium had the highest niche breadth and distributed in all ecosystem-types, while the genera with a narrow niche breadth distributed only in a few ecosystem-types. The results of niche overlap index analysis indicated that Stachybotrys and Torula, Doratomyces and Scolecobasidium, Cladosporium and Chrysosporium had a higher niche overlap, whereas Arthrinium and Gliomastix, Phialophora and Doratomyces, Oidiodendron and Ulocladium had no niche overlap.
China ; Ecosystem ; Fresh Water ; microbiology ; Mitosporic Fungi ; classification ; isolation & purification ; Soil Microbiology ; Species Specificity

We investigated the effects of fish protein hydrolysate (FPH) on growth performance and humoral immune response of the large yellow croaker (Pseudosciaena crocea R.). One thousand and two hundred large yellow croakers [initial average weight: (162.75+/-23.85) g] were divided into four groups and reared in floating sea cages (3 m x 3 m x 3 m). The animals were fed with 4 diets: basal diet only (control) or diets supplemented with 5%, 10% and 15% (w/w) FPH. The results show that dietary FPH levels significantly influenced the growth and immunity of the large yellow croaker. Compared with the control group, total weight gain (TWG) in all treatment groups, relative weight gain (RWG) and specific growth rate (SGR) in fish fed with diets supplemented with 10% and 15% FPH were significantly increased (P<0.05). Similar results were observed in immune parameters [lysozyme activity, serum complements, immunoglobulin M (IgM)]. Lysozyme activity, complement C4 and IgM were also significantly increased (P<0.05) in fish fed with diets supplemented with 10% and 15% FPH, while complement C3 level was significantly increased (P<0.05) in all treatment groups. In general, with the supplementation of FPH, particularly at dose of 10%, the growth performance and immunity of the large yellow croaker can be improved effectively.
Administration, Oral ; Animals ; Antibody Formation ; drug effects ; immunology ; Dietary Supplements ; Fish Products ; Gadiformes ; metabolism ; Perciformes ; growth & development ; immunology ; Protein Hydrolysates ; administration & dosage

OBJECTIVETo observe the role of theophylline in relieving airway symptoms and inflammation in patients with mild asthma.

METHODSFifty-six patients with mild asthma were randomly divided into treatment group (n=41) receiving oral theophylline at daily dose of 4 to 6 mg/kg for 16 weeks and control group (15 cases) without medication other than beta2 antagonist, which was administered when necessary in both groups. Peripheral blood T-lymphocyte subsets (CD3(+), CD4(+), and CD8(+)) and pulmonary function (PEF(am) and PD(20)) before and at 8 and 16 weeks during treatment were measured.

RESULTSSignificant difference was observed in CD3+ and CD4(+) T-lymphocyte subsets after medication with theophylline (P<0.05) in the patients, and PEF(am) and PD(20) were also significantly different from those of the control group (P<0.05). Theophylline significantly improved the clinical symptom scores (P<0.05) and decreased the asthma attacks.

CONCLUSIONLow-dose oral theophylline may significantly relieving airway inflammation in patients with mild asthma.

Administration, Oral ; Adult ; Asthma ; drug therapy ; immunology ; physiopathology ; Bronchodilator Agents ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Respiratory Function Tests ; T-Lymphocyte Subsets ; drug effects ; immunology ; Theophylline ; administration & dosage ; therapeutic use ; Treatment Outcome