AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-?B activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P

Objective To explore the feasibility of recently developed nucleofection method in delivering plamid DNA directly into the nucleus for the introduction of a plasmid encoding enhanced green fluorescent protein (EGFP) into primary bone marrow stromal cells (BMSCs) of rabbit. Methods Rabbit BMSCs were harvested by means of density gradient centrifugation following a thighbone puncture. The primary BMSCs were cultured and either transfected with pEGFP-C2 by nucleofector technology (as EGFP group) or uninfected (as control) in vitro. Compared with the control, the cellular viability, adhesive rates and the growth curves of the labeled cells were respectively analyzed. Transfection efficiencies were evaluated through the detection of EGFP expression. Results EGFP were successfully expressed 24 h after nucleofection. Similar morphological development, adhesive rates and growth curves were found in the 2 groups. The positive EGFP expression was enhanced gradually alone with the prolonged culture time, and showed the strongest 6 d after marked, with about 47.8% of EGFP-positive cells in the total BMSCs. The EGFP did not attenuate even 1 month after the marking. Conclusion Neuclofection of pEGFP-C2 shows no significant effect on the proliferation of rabbit BMSCs. EGFP plays an important role in stable gene marking of rabbit BMSCs. Nucleofection is an efficient nonviral gene transfer method for the introduction of genes into primary rabbit BMSCs.

Objective To investigate the clinical distribution and antimicrobial resistance of carbapenem-resistant Enterobacteriaceae (CRE) in Dongguan.Methods CRE isolated from hospitalized patients in 22 secondary and above medical institutions which participated in bacterial monitoring in Dongguan between January 2015 and June 2016 were retrospectively analyzed,antimicrobial resistance was analyzed by WHONET 5.6 software.Results A total of 71 CRE isolates were detected,with a isolation rate of 0.34％ (71/20 713).53 strains(74.65％) of CRE were isolated from patients aged 15-60 years old;46 (64.79 ％) were from male patients;CRE were mainly isolated from patients in intensive care unit(36 strains,50.70 ％);the main specimen was sputum(34 strains,47.89 ％),followed by urine (11 strians,15.49 ％) and wound secretion(6 strains,8.45 ％);the main infection type was healthcare associated infection (64 strains,90.14 ％);CRE were mainly distributed in tertiary hospitals(56 strains,78.87 ％),the isolation rate of CRE in tertiary and secondary hospitals were 0.41 ％(56/13 677)and 0.21％(15/7 036) respectively.71 strains of CRE were all resistant to imipenem,resistance rate to meropenem was 81.12％,only amikacin and tobramycin had drug resistance rates of ＜40％ (21.38％ and 38.79％ respectively),resistance rate to trime thoprim/sulfamethoxazole was 48.23 ％,while resistance rates to fluoroquinolones,third-generation cephalosporins,and enzyme inhibitors were all＞60.Conclusion The isolation rate of CRE in Dongguan is lower than that of the whole nation and the other provinces,effective prevention and control measures should be taken according to the key population and departments that isolated CRE,antimicrobial use should be rational.

Objective To investigate the photodynamic effect mediated with 5-aminolevulinic acid (5-ALA) on U251 human glioma cells. Methods Fluorescence microscope and confocal laser scanning microscope were used to detect the localization of Pp Ⅸ in U251 human glioma cells. The cells with/without 5-ALA were irradiated at the wavelength of 635 nm. MTT assay was used to measure the cell survival after laser irradiation. Results 5-ALA cocultured with U251 cells successfully produced endogenous Pp Ⅸthat was observed distributively in the cytoplasm, but not in nuclear region. The overall survival rates of the U251 glioma cells photodamaged by ALA-PDT decreased as the incubation time went by or the 5-ALA concentration increased, while peaked at the incubation time of 6 h and the 5-ALA concentration of 2.0mmol/L. Without one of 5-ALA and light irradiation, the survival rate of the cells had no significant difference compared with that of cells of the control group. Conclusion The 5-ALA-induced PDT appears to be a promising therapy for human glioma. The optimal incubation time may be 6 h and the optimal 5-ALA concentration be 2.0 mmol/L.

The purpose of study was to investigate the in vitro proliferation ability of PHA-induced CIK cells and traditionally prepared CIK cells, the effector cell level and its influence on killing activity to K562 cells, and to analyze the difference between them. The peripheral blood mononuclear cells(PBMNC) of healthy persons were isolated and divided into A and B group. The CIK cells in A group were obtained by using traditional culture method, the CIK cells in B group were prepared by PHA induction. During the cultivation, the cell survival rate and cell absolute value in the cell culture system were counted every 3 days. On day 15 of culture, the cell immunophenotype of 2 groups were detected by flow cytometry, and the ratios of CD3(+)CD56(+), CD3(+)CD8(+) and CD3(+)CD4(+) cells in total cell amount of culture system were accounted. Meantime, the killing activity to K562 cells in different effector-target ratios was detected by using CCK-8 kit between the 2 groups. The results showed that the method of preparing CIK by PHA induction promoted the cell proliferation more than that of the traditional method (P < 0.05), moreover, both the survival rate of cells in 2 groups was more than 90%. The CD3(+)CD8(+), CD3(+)CD56(+) cell ratio in 2 groups obviously increased. As compared with traditional method, the CD3(+)CD8(+) cell level in B group was enhanced (P < 0.05); but there were no statistical differences in increase of CD3(+)CD56(+) cell level and decrease of CD3(+)CD4(+) cell level between 2 groups. while the effector-target ratio is 5:1, 10:1, 20:1 and 40:1, the killing activity of PHA-induced CIK cells to K562 cells was more stronger than traditionally-prepared CIK cells (P < 0.05), moreover, along with increase of effector-target ratio, the difference of killing activity to K562 cells in 2 groups significantly increased. It is concluded that compared with traditional method for preparing CIK cells, the new way by PHA induction can increase the proliferation of CIK cells obviously, enhance the ratio of CD3(+)CD8(+) cells and strengthen the killing activity to the K562 cells. This new way provides a new source of CIK cells and reliable evidence for cyto-immune therapy of leukemia and other tumors.
Cell Proliferation ; Cytokine-Induced Killer Cells ; cytology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; Phytohemagglutinins ; pharmacology