BACKGROUND/AIMS: Transforming growth factor-β1 (TGF-β1) induction of epithelial-mesenchymal transition (EMT) is one of the mechanisms by which colorectal cancer (CRC) cells acquire migratory and invasive capacities, and subsequently metastasize. Parthenolide (PT) expresses multiple anti-cancer and anti-inflammatory activities that inhibit nuclear factor κB by targeting the IκB kinase complex. In the present study, we aimed to investigate whether PT can inhibit TGF-β1-induced EMT in CRC cell lines.METHODS: HT-29 and SW480 cell lines were used in the experiment. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and sub-G1 analysis was measured by flow cytometry. The induction of EMT by TGF-β1 and inhibition of the process by PT was analyzed by phase contrast microscopy, wounding healing, cellular migration and invasion assays, and Western blotting.RESULTS: TGF-β1 inhibits HT-29 cell proliferation, but has no effect on SW480 cell proliferation; different concentrations of TGF-β1 did not induce apoptosis in HT-29 and SW480 cells. PT attenuates TGF-β1-induced elongated, fibroblast-like shape changing in cells. PT inhibits TGF-β1-induced cell migration and cell invasion. In addition, other EMT markers such as β-catenin, Vimentin, Snail, and Slug were suppressed by PT, while E-cadherin was increased by PT.CONCLUSIONS: Our findings show that PT inhibits TGF-β1-induced EMT by suppressing the expression of the mesenchymal protein and increasing expression of the epithelial protein. These findings suggest a novel approach for CRC treatment by suppression of TGF-β1-induced EMT.
Apoptosis ; Blotting, Western ; Cadherins ; Cell Line ; Cell Movement ; Cell Proliferation ; Cell Survival ; Colorectal Neoplasms ; Epithelial-Mesenchymal Transition ; Flow Cytometry ; Gastropoda ; HT29 Cells ; Humans ; Microscopy, Phase-Contrast ; Phosphotransferases ; Snails ; Transforming Growth Factors ; Vimentin ; Wounds and Injuries

BACKGROUND: This study aimed to evaluate the adhesion of Acanthamoeba trophozoites on cosmetic contact lenses (CLs) with and without CL care multipurpose solution (MPS) treatment. METHODS: Acanthamoeba lugdunensis L3a trophozoites were inoculated onto disks trimmed from CLs: 1-day Acuvue moist, 1-day Acuvue define, Acuvue 2, and Acuvue 2 define. After 18-hour inoculation, the number of adherent trophozoites was counted under phase contrast microscopy. The effects of MPS, Opti-Free Express, soaking CLs for 6 hours, on Acanthamoeba adhesion were analyzed. Scanning electron microscopic examination was performed for assessment of Acanthamoeba attached on the lens surface. RESULTS: Acanthamoeba trophozoites showed greater adhesion to cosmetic CL (P = 0.017 for 1-day CL and P = 0.009 for 2-week CL) although there was no significant difference between the types of cosmetic CL. On all lenses, the number of adherent Acanthamoeba was significantly reduced after treatment with MPS (P < 0.001 for 1-day Acuvue moist, P = 0.046 for 1-day Acuvue define, P < 0.001 for Acuvue 2, and P = 0.015 for Acuvue 2 define), but there was still significant difference between conventional and cosmetic CLs (P = 0.003 for 1-day CL and P < 0.001 for 2-week CL, respectively). More attachment of Acanthamoeba was observed on colored area and the acanthopodia of Acanthamoeba was placed on the rough surface of colored area. CONCLUSION: Acanthamoeba showed a greater affinity for cosmetic CL and mostly attached on colored area. Although MPS that contained myristamidopropyl dimethylamine reduced the adhesion rate, there was a significant difference between conventional and cosmetic CLs.
Acanthamoeba ; Contact Lenses ; Microscopy, Phase-Contrast ; Trophozoites

This commentary presents the regulatory backgrounds and development of the national proficiency testing (PT) scheme on asbestos analysis in the Republic of Korea. Since 2009, under the amended Occupational Safety and Health Act, the survey of asbestos in buildings and clearance test of asbestos removal works have been mandated to be carried out by the laboratories designated by the Ministry of Employment and Labor (MOEL) in the Republic of Korea. To assess the performance of asbestos laboratories, a PT scheme on asbestos analysis was launched by the Korea Occupational Safety and Health Agency (KOSHA) on behalf of the MOEL in 2007. Participating laboratories are evaluated once a year for fiber counting and bulk asbestos analysis by phase contrast microscopy and polarized light microscopy, respectively. Currently, the number of laboratory enrollments is > 200, and the percentage of passed laboratories is > 90. The current status and several significant changes in operation, sample preparations, and statistics of assigning the reference values of the KOSHA PT scheme on asbestos analysis are presented. Critical retrospect based on the experiences of operating the KOSHA PT scheme suggests considerations for developing a new national PT scheme for asbestos analysis.
Asbestos* ; Employment ; Korea* ; Microscopy, Phase-Contrast ; Microscopy, Polarization ; Occupational Health ; Reference Values ; Republic of Korea

BACKGROUNDThis study characterized the cardiac telocyte (TC) population both in vivo and in vitro, and investigated its telomerase activity related to mitosis.

METHODSUsing transmission electron microscopy and a phase contrast microscope, the typical morphological features of cardiac TCs were observed; by targeting the cell surface proteins CD117 and CD34, CD117 + CD34 + cardiac TCs were sorted via flow cytometry and validated by immunofluorescence based on the primary cell culture. Then the optimized basal nutrient medium for selected population was examined with the cell counting kit 8. Under this conditioned medium, the process of cell division was captured, and the telomerase activity of CD117 + CD34 + cardiac TCs was detected in comparison with bone mesenchymal stem cells (BMSCs), cardiac fibroblasts (CFBs), cardiomyocytes (CMs).

RESULTSCardiac TCs projected characteristic telopodes with thin segments (podomers) in alternation with dilation (podoms). In addition, 64% of the primary cultured cardiac TCs were composed of CD117 + CD34 + cardiac TCs; which was verified by immunofluorescence. In a live cell imaging system, CD117 + CD34 + cardiac TCs were observed to enter into cell division in a short time, followed by an significant invagination forming across the middle of the cell body. Using a real-time quantitative telomeric-repeat amplification assay, the telomerase concentration in CD117 + CD34 + cardiac TCs was obviously lower than in BMSCs and CFBs, and significantly higher than in CMs.

CONCLUSIONSCardiac TCs represent a unique cell population and CD117 + CD34 + cardiac TCs have relative low telomerase activity that differs from BMSCs, CFBs and CMs and thus they might play an important role in maintaining cardiac homeostasis.

Animals ; Antigens, CD34 ; metabolism ; Fibroblasts ; enzymology ; ultrastructure ; Flow Cytometry ; Mesenchymal Stromal Cells ; enzymology ; ultrastructure ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Microscopy, Phase-Contrast ; Myocytes, Cardiac ; enzymology ; ultrastructure ; Proto-Oncogene Proteins c-kit ; metabolism ; Telomerase ; metabolism ; Vimentin ; metabolism

Resveratrol (trans-3,4'-trihydroxystillbene), a naturally occurring polyphenolic antioxidant found in grapes and red wine, elicits diverse biochemical responses and demonstrates anti-aging, anti-inflammatory, and anti-proliferative effects in several cell types. Previously, resveratrol was shown to regulate differentiation and inflammation in rabbit articular chondrocytes, while the direct production of nitric oxide (NO) in these cells by treatment with the NO donor sodium nitroprusside (SNP) led to apoptosis. In this study, the effect of resveratrol on NO-induced apoptosis in rabbit articular chondrocytes was investigated. Resveratrol dramatically reduced NO-induced apoptosis in chondrocytes, as determined by phase-contrast microscopy, the MTT assay, FACS analysis, and DAPI staining. Treatment with resveratrol inhibited the SNP-induced expression of p53 and p21 and reduced the expression of procaspase-3 in chondrocytes, as detected by western blot analysis. SNP-induced degradation of I-kappa B alpha (IkappaB-alpha) was rescued by resveratrol treatment, and the SN50 peptide-mediated inhibition of NF-kappa B (NF-kappaB) activity potently blocked SNP-induced caspase-3 activation and apoptosis. Our results suggest that resveratrol inhibits NO-induced apoptosis through the NF-kappaB pathway in articular chondrocytes.
Apoptosis* ; Blotting, Western ; Caspase 3 ; Chondrocytes* ; Humans ; I-kappa B Proteins ; Inflammation ; Microscopy, Phase-Contrast ; NF-kappa B* ; Nitric Oxide ; Nitroprusside ; Tissue Donors ; Vitis ; Wine

The objectives are to compare the airborne asbestos concentrations resulted from mitering of abestos cement roof sheets by a high-speed motor and a hand saw, and to monitor whether other workers near the test sites are vulnerable to the fibers exceeding the occupational exposure limit. Four test cases were carried out and altogether 7 personal and 4 area air samples were collected. The NIOSH method 7400 was employed for the air samplings and analysis. Using the phase contrast microscopy, fiber counting was conducted under Rule A. The study showed that the fiber concentration medians for personal air samples gathered from the two tools were 4.11 fibers/cc (ranged: 1.33-12.41 fibers/cc) and 0.13 fibers/cc (ranged: 0.01-5.00 fibers/cc) respectively. The median for the area samples was 0.59 fibers/cc (ranged: 0.14-3.32 fibers/cc). Comparing each study case, the concentration level caused by the high-speed motor saw was more than twice that of the hand saw. According to the area samples, the workers nearby the test site are at risk from high exposure to asbestos.
Asbestos ; Hand ; Humans ; Microscopy, Phase-Contrast ; National Institute for Occupational Safety and Health (U.S.) ; Occupational Exposure ; Organothiophosphorus Compounds

OBJECTIVETo compare the morphological difference between dermal tissue of normal skin and that of scar in rat, and to explore its structural pattern.

METHODSThe full-thickness skin and the scar tissue formed 3 weeks after wound healing from SD rats were harvested as samples, which were prepared appropriately afterwards. Samples were scanned and imaged with synchrotron radiation technology, micro-CT, and phase-contrast imaging technology. The images were rebuilt with three-dimensional software.

RESULTSThe micro-CT was materialized by using X-ray generated by synchrotron radiation light source. The structure of dermal tissues was clearly shown with the assistance of phase-contrast imaging technology in the process. It was demonstrated that the dermal tissues of normal skin of rat were mainly composed of collagenous fibers, which twined together to form an olive-like structure. These olive-like structures as basic units were arranged randomly in a certain way. The collagenous fibers in dermal tissue of the scar were arranged in a parallel manner, while some fibers were crooked and arranged in a disorderly manner.

CONCLUSIONSDermal tissue of normal skin in rat has stable three-dimensional structure, and its basic structure and manner of composition are obviously different from those of scar dermal tissue.

Animals ; Cicatrix ; diagnostic imaging ; Dermis ; diagnostic imaging ; pathology ; Imaging, Three-Dimensional ; methods ; Male ; Microscopy, Phase-Contrast ; Rats ; Rats, Sprague-Dawley ; Skin ; diagnostic imaging ; Synchrotrons ; Tomography, X-Ray Computed ; Wound Healing

OBJECTIVETo find an ideal method inducing dental pulp stem cells (DPSC) osteogenic differentiation. To compare the effect of co-culture method and that of mineralizing culture medium.

METHODSDPSC were co-cultured with osteoblasts using cell culture inserts system as experiment group, and DPSC were cultured in mineralizing culture medium as control group. The cell morphology and ultrastructure and mineralized nodes were analyzed under phase contrast microscope, transmission electron microscope, and alizarin red S staning. Bone sialoprotein (BSP), Runx-2, osteocalcin, and collagen-1 (Col-1) osteoblastic genes expressions of DPSC cultivated in special niche of osteoblasts were assayed by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe mineralization nudoles of experiment group were more than control group. Fifteen days later, BSP and Col-1 genes in the DPSC of co-cultures were 9.807 ± 1.135 and 2.913 ± 0.310, respectively. And those in the DPSC of mineralizing culture medium were 6.478 ± 0.781 and 1.703 ± 0.184, respectively. Co-cultures and mineralizing were significantly different (P < 0.05).

CONCLUSIONSAs osteoblasts can secret lots of osteogenic cell cytokines, they have more significant effect than mineralizing culture medium on osteogenesis of DPSC.

Cell Differentiation ; Coculture Techniques ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Dental Pulp ; cytology ; Gene Expression Regulation, Developmental ; Humans ; Integrin-Binding Sialoprotein ; metabolism ; Microscopy, Electron, Transmission ; Microscopy, Phase-Contrast ; Osteoblasts ; cytology ; Osteocalcin ; metabolism ; Osteogenesis ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; ultrastructure

OBJECTIVETo investigate the effect of bleomycin A5 on the hemangioma-derived endothelial cell line XTPS-1.

METHODSHemangioma-derived endothelial cell line XTPS-1 was cultured with different concentration of bleomycin A5 (1000, 100, 10, 1, 0 mg/L), and then the survival rate was measured by methyl thiazolyl terazolium (MTT), the variation of cell morphology was observed using inverted phase contrast microscope and electron microscope, the variation of cell cycle and apoptosis rate were measured using flow cytometry.

RESULTSAfter 24 hours culture the cell survival rate was (92.96 ± 3.66)% and (99.86 ± 0.12)% in lower saturation group (10 and 1 mg/L), but (34.08 ± 3.11)% and (43.28 ± 2.88)% in higher saturation group (1000 and 100 mg/L). The difference between them was more significant (P < 0.01). Lower saturation of bleomycin A5 (10 and 1 mg/L)could induce apoptosis but had almost no cytotoxic effect. Higher saturation of bleomycin A5 (1000 and 100 mg/L) not only induced apoptosis, but also had strong cytotoxic effect, which was concentration dependent.

CONCLUSIONSbleomycin A5 could induce apoptosis, inhibit cell proliferation and has direct cytotoxic effect.

Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Bleomycin ; administration & dosage ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Endothelial Cells ; pathology ; Hemangioma ; pathology ; Humans ; Microscopy, Electron, Transmission ; Microscopy, Phase-Contrast

PURPOSE: To investigate the effects of two antimetabolites, mitomycin C (MMC) and 5-fluorouracil (5-FU), on proliferation of cultured human nasal mucosa fibroblasts. METHODS: Human nasal mucosa fibroblasts were primarily cultured, and exposed to various concentrations of MMC and 5-FU for 5 minutes. Control fibroblasts were exposed to only DMEM media without the drugs. Effect of drugs on cell morphology was observed by phase-contrast microscopy. Cell viability and apoptosis were measured using MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide] assay and Acridine orange/Hoechst (AO/HO) staining, respectively. RESULTS: In both experimental groups exposed to MMC and 5-FU, fibroblasts maintained standard spindle shape. The MTT assay showed that both MMC and 5-FU inhibited fibroblast proliferation in a dose dependent manner. AO/HO staining showed apoptotic cells in both experimental groups. CONCLUSIONS: Both MMC and 5-FU have an antiproliferative effect on fibroblasts in vitro at least through induction of apoptosis. Therefore, adjuvant use of either MMC or 5-FU during endonasal dacryocystorhinostomy may improve the clinical outcome by inhibiting proliferation of the nasal mucosa.
Antimetabolites ; Apoptosis ; Cell Survival ; Dacryocystorhinostomy ; Fibroblasts ; Fluorouracil ; Humans ; Microscopy, Phase-Contrast ; Mitomycin ; Nasal Mucosa