ABSTRACT
Objective To investigate the prevalence of plasmid-mediated quinolone resistance(PMQR) in Proteus mirabilis clinical isolated from urine. Methods Antimicrobial susceptibility test was performed using the microdilution method.And PMQR gene qnrA,qnrB,qnrC,qnrD,qnrS,aac(6′)-Ib-cr,qepA were amplified by PCR,then the PCR positive products were sequenced to identify their genotypes. Results In 41 strains of Proteus mirabilis from urine,the resistant rates to ciprofloxacin and levofloxacin were 41.5% and 29.3%,respec-tively.Among all clinical isolates,qnrA1 gene was detected in 2 strains,qnrB2 gene in 3 strains.PMQR gene qn-rB,qnrC,qnrD,qnrS,aac(6')-Ib-cr and qepA were not detected in all strains.Conclusions Clinical isolates of Proteus mirabilis from urine carry PMQR genes.The prevalent principal genotypes are qnrA1 and qnrB2 in these iso-lates.They are related to low levels resistance toquinolone.
ABSTRACT
AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells.The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed.METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector.The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope.The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group.Non-transfected cells were used as control group.The cell transfection was carried out with 250 ng plasmids/well in 6-well plate.The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope.The mRNA expression of PKCε was detected by RT-qPCR.The protein expres-sion of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot.The cell proliferation ability was detec-ted with colony formation assay.The cell invasion ability was detected by Transwell method.RESULTS: The expression of CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining.The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76 ± 0.18)%, (98.51 ±0.32)%, (99.17 ±0.16)% and (99.68 ±0.11)%, respectively.The difference between CP group and control group was statistically significant (P <0.05).No significant difference among CN group, LP group and control group was observed.The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference.The colony number in CP group was significantly smaller than that in control group (P <0.05).The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group.The number of the invading cells in CP group was significantly less than that in control group (P <0.05).The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group.CONCLUSION: Nanogene vector targe-ting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibito-ry effects on the proliferation and invasion of the lung cancer cells.
ABSTRACT
AIM:To explore the effects of decorin on procollagen type I (PcI), mRNA expression,collagen type I synthesis and proliferation of synovial type B cells of stiff knee joint synovial membrane.METHODS:Type B cells of synovial membrane were isolated from the stiff knee joint synovial membrane and cultured in vitro.The cells were treated with decorin at concentrations of 0.1 mg/L, 5 mg/L and 10 mg/L.After cultured for 24 h, 48 h and 72 h, the cell proli-feration rates were measured by MTT colorimetric determination.Cell cycle distribution and apoptosis were analyzed by flow cytometry.The mRNA level of Pc I was detected by RT-PCR, while collagen type I was measured by Western blot.RE-SULTS:The proliferation of synovial type B cells was significantly inhibited, the percentage of synovial type B cells at G1 phase was significantly increased by 5 mg/L and 10 mg/L decorin (P<0.05), and PcⅠmRNA expression and collagen type I synthesis were significantly decreased.The cells with late apoptosis were not found in control group and experimental groups.CONCLUSION:Recombinant human decorin inhibits synovial type B cell proliferation and decreases PcⅠmRNA expression and collagen type I synthesis in synovial type B cells of stiff knee joint synovial membrane in vitro, suggesting that decorin potentially contributes to the therapy of human knee stiffness.