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.

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

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

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

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

Foodbome viruses are defined to be viruses that can lead to human diseases through food. In accordance with the different origin, foodborne viruses can be divided into two kinds: intestinal viruses and zoonotic viruses. The former include those viruses that can be transmitted to person via fecal-orally route. The latter include those zoonotic viruses that chiefly transmitted to person through livestock and poultry products. This paper expounds foodborne viruses categories, biology nature, epidemiology character, and study circumstance, and clarifies the molecular biological methods and problems on the base of the polymerase chain reactions, and presents the development direction and application perspective of the foodbome viruses study.

In this study, three methods for identification of E.coli were compared. The conventional method was employed to select and identify the suspicious E.coli isolates from a fecal sample. PCR based ARDRA analysis was then carried out to distinguish these E.coli isolates, E.coli MG1655 and other bacterial species. All the potential E.coli isolates and E.coli MG1655 had the identical ARDRA banding pattern while the other bacterial species showed the different patterns.The result indicated that the ARDRA analysis was consistent with the traditional method for identification of E.coli and could be the practical method for distinguishing E.coli from other intestinal bacterial species. The ERIC-PCR analysis provided abundant polymorphism between different E.coli isolates, and might be a powerful approach for elucidating the genetic diversity among isolates of the same species.

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.

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.