In the previous review, the physical aspect of heavy particles, with a focus on the carbon beam was introduced. Particle beam therapy has many potential advantages for cancer treatment without increasing severe side effects in normal tissue, these kinds of radiation have different biologic characteristics and have advantages over using conventional photon beam radiation during treatment. The relative biological effectiveness (RBE) is used for many biological, clinical endpoints among different radiation types and is the only convenient way to transfer the clinical experience in radiotherapy with photons to another type of radiation therapy. However, the RBE varies dependent on the energy of the beam, the fractionation, cell types, oxygenation status, and the biological endpoint studied. Thus this review describes the concerns about RBE related to particle beam to increase interests of the Korean radiation oncologists' society.
Carbon ; Cell Fractionation ; Oxygen ; Photons ; Population Characteristics ; Protons ; Relative Biological Effectiveness

The schemes of dose fractionation play an important role in tumor radiotherapy. We used mathematical methods to describe the process of tumor cells evolution during radiotherapy, trying to find how the schemes of dose fractionation affect tumor cells. In clinical radiobiology, linear-quadratic (LQ) model is frequently used to describe radiation effects of tumor cells. We integrated LQ model with effect of oxygen, and with the phenomenon of repopulation and reoxygenation in the theory of radiation biology. While we considered the disappearing progress of doomed cells in tumor, we established the mathematical model of tumor evolution in radiotherapy. We simulated some common treatment schedules, and studied the change role of tumor cells during radiotherapy. These results can serve for the optimization of dose fractionation scheme based on tumor radiobiological characteristics.
Cell Growth Processes ; radiation effects ; Dose Fractionation ; Humans ; Models, Theoretical ; Neoplasms ; pathology ; physiopathology ; radiotherapy ; Radiobiology

Being an essential component of systematic biology, metabolomics has received attention in recent years. It is a post genomic technology aimed at qualitative and quantitative analysis of all low molecular-mass metabolites present in complex biological samples, and mainly investigates the change of endogenous metabolites of a stimulated or disturbed biological system. Investigations into intracellular endogenous metabolites in metabolomics have great advancement in recent years. This review outlines the progress of metabolomics in cell culture analysis including sample preprocessing methods and metabolite target analysis, metabolic profiling analysis, metabolomics analysis and metabolic footprinting analysis.
Animals ; Cell Culture Techniques ; Chemical Fractionation ; methods ; Humans ; Intracellular Space ; metabolism ; Metabolome ; Metabolomics ; methods

OBJECTIVETo establish a technology platform for the preparation of interphase nuclei for the fluorescence in situ hybridization (FISH) detection of solid tumor tissues.

METHODSThe centromere probe of chromosome 3 was labeled by the random primer technique, and then hybridized to interphase nuclei prepared by six different methods in order to study the influence on FISH detection.

RESULTSEach method of slide preparation had its own characteristic, and could be used according to different needs. As regards to FISH, collagenase method got the best results. Whereas for frozen samples or small tissues, to prepare printing slides was more applicable.

CONCLUSIONThe comparison of different slide preparation methods lays a technology foundation for the FISH application in cancer researches and clinical diagnosis of solid tumors.

Animals ; Cell Fractionation ; methods ; Cell Nucleus ; metabolism ; Collagenases ; metabolism ; In Situ Hybridization, Fluorescence ; methods ; Interphase ; Neoplasms ; genetics ; pathology

OBJECTIVETo analyse the result of late course accelerated hyper-fractionated radiotherapy (LCAHFR) of upper and middle thoracic segment esophageal T2N0M0 carcinoma.

METHODSFifty-three patients with squamous cell esophageal T2N0M0 carcinoma in the upper and middle segment were treated by LCAHFR from August 1994 to January 2000. The design of the radiation fields were based on CT and barium examination. All patients were treated with the conventional fractionated radiotherapy during the first two-thirds of the treatment to a dose about 41.4 Gy/23 F/4 to 5 weeks. This was followed by accelerated hyper-fractionated irradiation using reduced fields, twice daily at 1.5 Gy per fraction to a dose about 27 Gy/18 d. Thus, the total dose was 67-70 Gy/40-43 F/40-49 d.

RESULTSThe 1-, 2- and 5-year actuarial survival rates were 89.9%, 66.8% and 51.2%, respectively. The 1-, 2- and 5-year local control rates were 92.1%, 87.1% and 87.1%. Of the 17 patients who died, 5 died of local failure (29.4%), 9 (52.9%) of distant metastasis, 5 (29.4%) of lymph metastasis and 1 (5.9%) of bleeding from the esophagus. The Cox multivariate model showed that the site of lesion was the only prognostic factor, with upper better than the middle segment.

CONCLUSIONLate course accelerated hyper fractionated radiotherapy is one of the best radiation treatment regimen for early esophageal carcinoma in the upper and middle thoracic segment.

Aged ; Carcinoma, Squamous Cell ; radiotherapy ; Dose Fractionation ; Esophageal Neoplasms ; radiotherapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo extract the plasma membrane proteins from mouse liver cells and investigate the approach for fractionating the protein mixtures by two-dimensional liquid chromatography.

METHODSThe plasma membrane of the liver cells from 10 mice was extracted by differential centrifugation and sucrose density-gradient centrifugation. The plasma membrane proteins were exchanged with the start buffer and separated by chromatofocusing in the first-dimensional fractionation. The final results were transformed into UV/pI maps using ProteoVue software.

RESULTSWe successfully extracted the plasma membrane proteins from mouse liver cells. Sixteen fractions between pH 8.5-4.0 were recovered in the first-dimensional chromatofocusing followed by 2D- chromatographic fractionation, and the results were displayed as UV/pI maps.

CONCLUSIONThis approach for fractionating the mouse liver cell plasma membrane protein study provides the foundation for further studies on the functions of plasma membrane proteins and differential proteome of diseases.

Animals ; Cell Membrane ; metabolism ; Chemical Fractionation ; instrumentation ; methods ; Chromatography, Liquid ; methods ; Liver ; cytology ; Male ; Mice ; Proteome ; metabolism ; Proteomics ; instrumentation ; methods

Liquid-Based Uterine Cervicovaginal Cytology is known to be a sensitive and effective screening method for cervical neoplasm. MonoPrepTM, ThinPrepTM, and SurePathTM methods have been recently used as Liquid- Based Uterine Cervicovaginal Cytology techniques, and the SurePathTM method has been used in Sung-Yoon Reference Laboratory since 2003. The goal of Liquid-Based Uterine Cervicovaginal Cytology is to separate cervical epithelial cells from non-target cells, red blood cells and neutrophils. This report describes a study which evaluated cellularity, stainability, and cellular changes of epithelial cells in samples processed using a manual technique as compared to samples processed using SurePathTM automated method. The samples processed by means of a manual technique contained a cellularity of epithelial cells similar to that of the samples processed using the SurePathTM automated method. In addition, we compared variable density gradient reagents, including dextran, dextrose, and sucrose, to SurePathTM gradient media in order to evaluate cell fractionation and cellularity of epithelial cells. 10% dextran of gradient media shows good fractionation. The samples processed with 10% dextran demonstrated sufficient cellularity of epithelial cells and shows the fewest cellular changes. In conclusion, using a manual technique on these samples is easier to read than those results obtained using the SurePathTM automated method.
Cell Fractionation ; Cervix Uteri ; Dextrans ; Epithelial Cells ; Erythrocytes ; Female ; Glucose ; Indicators and Reagents ; Mass Screening ; Neutrophils ; Sucrose ; Uterine Cervical Neoplasms

PURPOSE: Previously, we reported that the expression of E-cadherin was significantly decreased according to the increase of the level of hepatocyte growth factor (HGF) in gastric cancer tissue. In this work, the effect of HGF on the cell-cell adhesion and intracellular distribution of E-cadherin in the gastric carcinoma cell lines were studied. MATERIALS AND METHODS: Western blot analysis was performed to confirm the presence or abscence of c-Met and E-cadherin in SNU-1, 5, and 16 cells. Tyrosine phosphorylation of c-Met, E-cadherin, alpha-, beta-, gamma-catenins was checked by immunoprecipitation. The morphologic changes induced by HGF were studied with immunocytochemical staining. Functional proportion of E-cadherin was estimated by cell fractionation. The effect of HGF on cell proliferation and invasion was also assessed. RESULTS: Among SNU-1, 5, and 16 cell lines, only SNU-16 cells expressed both E-cadherin and c-Met. A morphological change from epithelial shape to fibroblastic one was observed in the SNU-16 cells after treatment with HGF. In addition, E-cadherin expression of the SNU-16 cells was shifted from the membrane and to the cytoplasm, and the functional fraction of E-cadherin was decreased in the SNU-16 cells treated with HGF. On the other hand, HGF increased the proliferation and invasion of the SNU-16 cells. CONCLUSION: These results suggest that HGF may regulate cell adhesion in gastric carcinomas via the cellular redistribution and functional change of E-cadherin.
Blotting, Western ; Cadherins* ; Cell Adhesion ; Cell Fractionation ; Cell Line* ; Cell Proliferation ; Cytoplasm ; Fibroblasts ; gamma Catenin ; Hand ; Hepatocyte Growth Factor* ; Hepatocytes* ; Immunoprecipitation ; Membranes ; Phosphorylation ; Stomach Neoplasms ; Tyrosine

Cell-free protein expression system is a new method to express target protein in vitro and has been widely applied to the study of protein structure, protein function and other related fields. Preparation of cell extract is one of the key factors that affect the efficiency of the cell-free system. To improve the efficiency and economical feasibility of cell-free protein synthesis, we discussed the parameters during the preparation of the cell extract. These parameters include centrifugation speed, pre-incubation, and dialysis. We used the green fluorescent protein as the reporter protein, and obtained a simple procedure for the preparation of Escherichia coli cell extract. A simple centrifugation step (12 000 x g, 10 min) followed by a brief incubation was sufficient for the preparation of an active cell extract to support protein expression with higher productivity (209 microg/mL). Compared to the traditional E. coli S30 procedure, the processing time was reduced by 62%, and the productivity was increased by 2.6 times. The new procedure will make the advantage of cell-free technology more obvious, and promote its wider application.
Cell Fractionation ; methods ; Cell-Free System ; Escherichia coli ; cytology ; genetics ; metabolism ; Escherichia coli Proteins ; biosynthesis ; chemistry ; isolation & purification ; Green Fluorescent Proteins ; metabolism

OBJECTIVES: The authors would report the results of definitive radiation therapy (RT) for early glottic cancer by two different radiation dose schedules. METHODS: From February of 1995 till June of 2008, 157 patients with T1-2N0 glottic cancer were treated with curative RT at Samsung Medical Center. All patients had squamous cell carcinoma, and there were 89 patients (56.7%) with T1a, 36 (22.9%) with T1b, and 32 (20.4%) with T2. Two different radiation dose schedules were used: 70 Gy in 35 fractions to 64 patients (40.8%, group A); and 67.5 Gy in 30 fractions to 93 patients (59.2%, group B). The median treatment durations were 50 days (range, 44 to 59 days) and 44 days (range, 40 to 67 days) in the groups A and B, respectively. RESULTS: The median follow-up durations were 85 and 45 months for the groups A and B. No severe late complication of RTOG grade 3 or higher was observed, and there was no difference in acute or chronic complication between the groups. Twenty-four patients experienced treatment failure: local recurrence only in 19 patients; regional recurrence only in one; combined local and regional recurrence in four; and systemic metastasis in none. The overall 5-year disease-free survival and disease-specific survival rates were 84.7% and 94.8%. The disease-free survival rate in the group B was better (78.3% vs. 90.8%, P=0.031). This difference was significant only in T1 stage (83.4% vs. 94.6%, P=0.025), but not in T2 (62.7% vs. 60.6%, P=0.965). Univariate analysis showed that the tumor extent, cord mobility, T-stage, and the dose schedule had significant influence on the disease-free survival, and multivariate analysis showed that only the tumor extent and the dose schedule were associated with the disease-free survival. CONCLUSION: Superior disease-free survival could be achieved by 2.25 Gy per fraction without increased toxicity over shorter RT duration, when compared with 2.0 Gy per fraction.
Appointments and Schedules ; Carcinoma, Squamous Cell ; Disease-Free Survival ; Dose Fractionation ; Follow-Up Studies ; Humans ; Laryngeal Neoplasms ; Multivariate Analysis ; Neoplasm Metastasis ; Recurrence ; Survival Rate