OBJECTIVETo determine the interaction between miR-21 and DNA methylation in different breast cancer cells.

METHODSFluorescence tagged miR-21 inhibitor and its negative control (NC) were transient transfected into MCF-7 and MDA-MB-231 cell, the transfection efficiency was observed using fluorescence microscopy, and the miR-21 expression level and genome DNA methylation status before and after transfection were assessed by real-time PCR and bisulfite-qMSP respectively. To investigate the regulation effect of DNA methylation on miR-21, cells were treated with 5-AZA (2.5 µmol/L) for 72 h, with dimethyl sulfoxide (DMSO) treatment as its negative control (NC), and the expression level of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and AKT(also known as Protein Kinase B), two downstream genes of miR-21 were detected by Western blot.

RESULTSThe expression of miR-21 in MCF-7 cell was significantly knocked down (P < 0.01) by miR-21 inhibitor, with the genome DNA methylation level (P < 0.05) and all the three Dnmts: Dnmt1, Dnmt3a, and Dnmt3b unregulated. In contrast, the miR-21 expression in MDA-MB-231 cell was elevated ( P < 0.01) by miR-21 inhibitor, meanwhile, down- regulated of genome DNA methylation (P < 0.05) and Dnmt3b expression, upregulation of Dnmt3a were also observed. In addition, treated with 5-AZA resulted in significant increases of miR-21 expression in both MCF-7 and MDA-MB-231 cells (P < 0.01), with the protein level of PTEN increased in MCF-7 cell, which was further involved in the downregulation of AKT.

CONCLUSIONThe regulation effects of DNA methylation by transient transfection of miR-21 in MCF-7 and MDA-MB-231 cells are almost opposite, whilst the expression of miR-21 in two cell lines were all upregulated by decreased DNA methylation level and our results may provide some experimental evidences for the future development of rational therapy for different breast cancer.

Azacitidine ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MCF-7 Cells ; MicroRNAs ; genetics ; PTEN Phosphohydrolase ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins c-akt ; metabolism ; Real-Time Polymerase Chain Reaction ; Up-Regulation

AIM:To observe the clinical effects of 23-gauge (23G) transconjunctival sutureless vitrectomy for idiopathic macular hole.
METHODS: In this retrospective study, 28 eyes of 28 consecutive patients who underwent 23 - gauge transconjunctival sutureless vitrectomy for idiopathic macular hole between January 2013 and October 2013 in our hospital were evaluated. The follow-up time was 3-12mo. The operation effects were analyzed.
RESULTS:The macular hole was closed in 27 eyes of 28 eyes which underwent 23G transconjunctival sutureless vitrectomy and not closed in 1 eye after surgery. Best-corrected visual acuity at postoperative 1, 3mo was significantly improved compared to pre-operation (χ2=8-65, P=0. 003;χ2=10. 33, P=0. 001). The macular hole was closed as shown by OCT. Intraoperative incision was sutured in 5 cases ( 18%) . There was no statistically significant difference in intraocular pressure between pre-operation and post - operation. No post - operative complications such as endophthalmitis, retinal detachment, vitreous hemorrhage came up.
CONCLUSION: 23G transconjunctival sutureless vitrectomy is observed to be safe and effective technique in the treatment of macular hole. It is therefore our preferred system for straightforward macular surgery.

Objective To separate the side population cells(SP) from breast cancer MCF-7 cell line,and observe its biological characteristics.Methods Flow cytometry and Hcechst 33342 dye efflux assay were used to isolate SP cells and non-SP cells from the MCF-7 cell line of human breast cancer.Tumorigenicity of the two subpopulations was observed by a soft agar cloning method.Results The results of FACS analysis indicated that (6.5 ± 0.4 ) ％of the MCF-7 cells were SP cells;The vitro colony formation rate of SP cells was(38.5 ±9.4)％,and higher than that of non-SP cells ( 8.4 ± 2.6 ) ％ ( t =5.34,P ＜ 0,05 ).Concluslon The SP cells sorted from MCF-7 cell line enriched tunor stem cells,which exhibited high tumorigenicity.It indicated that SP cells should play a principal role in breast cancer.

Objective To study the metabolic change of proton magnetic resonance spectroscopy (1 H-MRS)in the visual cortex and optic radiation region of patients with diabetic retinopathy (DR).Methods 1H-MRS was performed in 20 patients with DR and 20 healthy volunteers on GE 1.5 T MR system respectively.Metabolic peaks of N-acetylasparte(NAA),creatine(Cr,in 3.02 and 3.94 ppm),choline-containing compounds(Cho)and myo-inositol(mI)were observed,and the ratios were analyzed by each other.Independent-samples t test was performed between two sets of data.Results In both visual cortex and optic radiation,the ratios of ml/Cr and mI/CrSee in DR group(0.664 ± 0.052 and 1.453±0.068 in visual cortex,0.717±0.074 and 1.484±0.114 in optic radiation)were significant higher than those in normal group(0.602±0.047 and 1.249±0.044 in visual cortex,0.679 ± 0.075 and 1.334±0.089 in optic radiation,P＜0.05).In DR group,the ratios of CrSec/Cr,Cho/Cr and NAA/Cr in visual cortex and optic radiation were 0.458±0.043 and 0.488±0.052,0.481±0.057 and 0.807±0.110,1.633±0.105 and 1.709±0.140 respectively.In control group.the ratios of those were 0.484±0.041 and0.502±0.056,0.471±0.065 and 0.786±0.109,1.625±0.098 and 1.716±0.135 respectively.The ratios of CrSec/Cr,Cho/Cr and NAA/Cr had no statistic difference between two groups(P＜0.05).Conclusion mI/CrSec is a typical change in the visual cortex and optic radiation region,1H-MRS as a noninvasive examination could provide biochemical and metabolic informations for diabetic patients.

Abdominal Cavity ; physiopathology ; Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Objective To study and compare the protective effects of pmtease inhibitor and corticosteroid on endotoxin-indueed acute lung injury in order to guide the choice of appropriate drugs. Method Thirty-two New Zealand rabbits were randomly divided(random number) into four groups with 8 rabbits in each, namely normal controls(C) ; lipopolysaecharide(LPS) group(L) ; ulinastatin(UTI) group(U) and dexamcthasone(DEX) group (D) .Except group C, all rabbits were injected with a dose of LPS 600 μg/kg iv. Meanwhile the rabbits in group U,group D received UTI(100 000 μ/kg), DEX(5 mg/kg), respectively. The specimens were collected 4 hours later for detecting the levels of TNF-α and NO in serum, and blood gas analysis, histological manifestations, the lung wet/dry weight ratio, lung tissue MPO and SOD activity, lung tissue MDA. Data were analyzed by ANOVA (SNK- q test), and P < 0.05 was considered as significantly different. Results Compared with group C, the lungs of the rabbits in group L had inflammatory granulocyte infdtration, diffused alveolar septum thickening and hemorrhagic spots were observed in pathological examinations. The histological changes of group U and group D were much lessened than those in group L. As groups U and D were compared with group L, there were significant differences inmany biomarkers including lung wet/dry weight ratio[(5.02±0.11),(4.93±0.13) vs.(5.37 ±0.29)],lung tissue MPO activity[(0.51 ± 0.05),(0.54±0.07) vs.(0.82 ± 0.09)] and MDA[(0.82 ±0.05),(0.81 ±0.04) vs.(0.96±0.05)], NO[(296.2± 11.7),(291.7 ± 15.8) vs.(351.8±19.6)] and TNF-α[group D(2.021 ± 0.122) vs. group L(4.999 ± 0.139)],lung tissue SOD activity[(120.3 ± 6.1),(122.6±3.5) vs.(105.1 ± 8.5)] and blood gas analysis[pH(7.30±0.23),(7.30±0.17) vs.(7.22±0.45) and PaO_2( 101.9 ± 6.8).( 102.5 ± 4.7) vs.(80.3 ± 3.3)] ; but there were no differences of above mentioned biomarkers between group U and D( P > 0.05). And there were no significant differences in PaCO_2 betweeu group U and D and group L[(37.0 ± 3.3),(37.6 ± 3.0) vs.(34.8 ± 2.3)]( P > 0.05). Conclusions The protective effects of ulinastatin on endotoxin-induced acute lung injury is comparable to those of dexamethasone, thus the former may be a clinical substitute for the latter with less side effects.

Adult ; Case-Control Studies ; Comet Assay ; DNA Mutational Analysis ; Female ; Humans ; Male ; Micronucleus Tests ; Middle Aged ; Occupational Exposure ; Vincristine

OBJECTIVETo study the effect of Ti-TiO2 nanotubes with different diameters on the adhesion of fibroblast and osteoblast, and to find which diameter was more favorable for cells' respective adhesion.

METHODSPure titanium sheets were polished and then anodized at different potentials for 1 h with Ti as anode and Pt as cathode. TiO2 nanotubes formed at 1, 5, 10 and 20 V potentials served as experimental groups and polished pure titanium served as control group. Field emission scanning electron microscopy (Fe-SEM) was used to analyze the surface topography. Stained nucleus with Hoechst33342 were used to measure the cell adhesion. The cell shape on the sample surface were analyzed with Fe-SEM.

RESULTSTiO2 nanotube array of different inner diameters from 15 nm to 100 nm were grown on titanium sheets by anodization at potentials from 1 to 20 V. At 30, 60 and 120 min, fibroblast adhesion at nanotubes anodized at 5 V was (141 ± 9), (388 ± 14) and (489 ± 15) respectively, significantly less than any other nanotube surface at the same time (P < 0.01). Nanotubes anodized at 20 V had the least inhibitory effect for fibroblast adhesion with a number of (579 ± 14) at 120 min, and the cell shape was also inhibited. At 30, 60 and 120 min, osteoblast had a significant better adhesion on nanotubes formed at 5 V than it did on any other surface at the same time (P < 0.01), except the control group at 30 min, with the adhesion number of (198 ± 10), (431 ± 10) and (501 ± 10) respectively, and osteoblast had a abundant spread on nanotubes formed at 5 V; while osteoblast adhesion on nanotubes anodized at 20 V was (152 ± 11), (403 ± 9) and (465 ± 12) respectively, less than on any other nanotube surface within the same time (P < 0.05), and the cell shape on the surface changed to be more elongate.

CONCLUSIONSFibroblast adhesion is inhabited more or less on Ti-TiO2 nanotubes of different diameters. Nanotubes formed at 5 V have the most osteoblast adhesion, and inhibit fibroblast adhesion.

Animals ; Cell Adhesion ; Fibroblasts ; cytology ; ultrastructure ; Mice ; Microscopy, Electron, Scanning ; Nanotubes ; chemistry ; Osteoblasts ; cytology ; ultrastructure ; Surface Properties ; Titanium ; chemistry

Environmental Monitoring ; methods ; Inhalation Exposure ; Occupational Exposure ; analysis ; Workplace

BACKGROUND: Anterior and dorsal root nerve tracts should be separated to small tracts in high selective dorsal rhizotomy, because detailed tract separation will benefit electrostimulation, thereby helping correctly cutting the lower-threshold Ia nerve fibers that cause convulsion, and meanwhile sensory nerve fibers in dorsal nerve root can be reserved as many as possible.OBJECTIVE: To meet the needs of limited and high selective spinal dorsal rhizotomy, anterior and dorsal root of spinal nerve were microanatomized to be certain of the separation standard and the number of small nerve tracts, so as to provide reliable basis and novel operative standard for clinical operation.DESIGN: Single sample experiment with adult corpses as subjects.SETTING: Orthopedic Department of the First Affiliated Hospital and Department of Anatomy, Jinzhou Medical College.PARTICIPANTS: This study was carried out at the Anatomical Laboratory of Jinzhou Medical College in December 1999. Fifteen adult corpses, 11 males and 4 males, were donated, and the donators signed informed consent when alive.tained from the 15 adult spinal cords (30 sides) and subjected to morphoanterior and dorsal roots of L5 spinal cord were obtained from a fresh corpse for immunohistochemical staining. The starting part, middle part and the exterior of intervertebral foremen was cut into slices, and the total number of nerve fibers, the number of Ia nerve fibers responsible for convulsion, and their percentage in the total fibers were counted. Meanwhile the distribution and the number of Ia nerve fibers in the three parts were compared.ber of nerve fibers per 100 μm2, the percentage of Ia nerve fibers in the total nerve fibers at the starting part, middle part and exterior of intervertebral foremen of spinal nerve dorsal root.root filaments. Microsurgical observation proved that dorsal root could be divided into 10-18 small tracts and anterior root 6-11 tracts; the diameter number of nerve fibers in the three parts of spinal nerve dorsal root was (3 243±143) fibers per 100 μm2, including (1 702±85) Ia nerve fibers that constituted about 52.5% of the total nerve fibers. Ia nerve fibers were found to be evenly distributed in dorsal root and no gathering could be observed.CONCLUSION: The tracts in anterior and dorsal roots of spinal cord should be separated as minutely as possible during improved dorsal rhizotomy, which is beneficial for cutting off Ia nerve fibers correctly. Generally,the anterior root consists of 6-11 small tracts and dorsal roots of 10-18small tracts, and nerve tracts should not be cut off over 1/2 of total dorsal nerve fibers.