Polymerase chain reaction (PCR) has been used as a substitute for conventional serological methods in order to provide blood or blood products free from contaminating viruses and recently attempts have focused to detect 2 or 3 viruses by a single multiplex PCR (M-PCR) reaction. We were able to detect human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), hepatitis B virus (HBV) and human cytomegalovirus (HCMV) simultaneously by a single M-PCR. However detection by gel electrophoresis of the products from M-PCR suffers from drawbacks such as low sensitivity and product sizes. Here we report enhanced detection systems of M-PCR based on nucleic acid hybridization with arrays built on membrane. Membrane array was manufactured by spotting appropriate probe DNAs on nylon membrane. Single or multiplex PCR was performed and the PCR products were labeled with DIG and allowed to hybridize with the membrane array. Results indicate that nonspecific hybridization was not observed for membrane DNA array. Additionally, membrane array method could detect small amount of viruses that were not detectable by conventional gel electrophoresis. At least 25-fold, and in some cases more than 125-fold increases in sensitivity was obtained with DNA array method. Thus, the nucleic acid hybridization with membrane array could be applied for the detection of M-PCR of viruses in blood or blood products.
Cytomegalovirus ; DNA ; Electrophoresis ; Female ; Hepacivirus ; Hepatitis B virus ; HIV-1 ; Humans ; Membranes ; Metrorrhagia ; Multiplex Polymerase Chain Reaction* ; Nucleic Acid Hybridization* ; Nylons ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction

BACKGROUND/AIMS: Enhanced replication of hepatitis C virus (HCV) is well described in the setting of moderate to severe immunosuppression. The aims of this retrospective study were to determine the incidence of enhanced HCV replication in hepatocellular carcinoma (HCC) patients undergoing transarterial chemolipiodolization (TACL) and to identify the factors associated with enhanced replication of HCV. The clinical pattern of enhanced HCV replication was compared with hepatitis B virus (HBV) reactivation during TACL. METHODS: This study enrolled 49 anti-HCV-seropositive patients who were diagnosed with HCC between January 2005 and December 2010 and who underwent TACL using epirubicin and/or cisplatin with consecutive HCV RNA copies checked. For comparison, 46 hepatitis B surface antigen1-positive patients with HCC who were treated with TACL were also enrolled. The frequency, associated factors, and clinical outcomes of enhanced HCV replication were analyzed and compared with those of HBV reactivation during TACL. RESULTS: Enhanced replication of HCV occurred in 13 (26.5%) of the 49 anti-HCV-seropositive patients during TACL. Of these 13 patients, 4 developed hepatitis, but none of the subjects developed decompensation due to the hepatitis. No significant clinical factors for enhanced HCV replication during TACL were found. Compared with HBV reactivation, the frequency of hepatitis attributed to enhanced HCV replication was significantly lower than that for HBV reactivation (8.2% vs. 23.9%, P=0.036). CONCLUSIONS: TACL can enhance HCV replication; however, the likelihood of hepatitis and decompensation stemming from enhanced HCV replication was lower than that for HBV reactivation in patients undergoing TACL.
Adult ; Aged ; Antineoplastic Agents/*administration & dosage/adverse effects/pharmacology ; Carcinoma, Hepatocellular/complications/*therapy ; Chemoembolization, Therapeutic/*adverse effects ; Drug Therapy, Combination ; Female ; Hepacivirus/drug effects/*physiology ; Hepatitis B/complications/epidemiology/virology ; Hepatitis B Surface Antigens/blood ; Hepatitis B virus/drug effects/*physiology ; Hepatitis C/complications/epidemiology/virology ; Humans ; Liver Neoplasms/complications/*therapy ; Male ; Middle Aged ; RNA, Viral/analysis ; Retrospective Studies ; Virus Activation ; *Virus Replication

Natural killer (NK) cells play an important role in innate immunity, especially in the response to viral infections, such as hepatitis C virus (HCV). Killer cell immunoglobulin-like receptors (KIRs) are the primary receptors of NK cells that mediate innate immunity. KIRs are also involved in acquired immunity, because some KIRs are expressed on the surface of certain subsets of T cells. In this study, the frequency of KIR genes, HLA-C allotypes, and combinations of KIR genes with their HLA-C ligands were evaluated in two different groups of the Korean population: controls and patients with chronic HCV infection. The study population consisted of 147 Korean patients with chronic HCV infection. The frequency of KIR2DS2 in patients with chronic HCV infection was 9.5% which was significantly lower than 19.5% of the control (P < 0.01). However, there were no significant differences in the frequency of other KIR genes, HLA-C allotypes or different combinations of KIR genes with their HLA-C ligands. This study can contribute to the further prospective study with a larger scale, suggesting the assumption that KIR2DS2 might aid in HCV clearance by enhancing both the innate and acquired immune responses of people in Korea.
Adult ; Aged ; Female ; Genes, MHC Class I ; Genotype ; HLA-C Antigens/genetics ; Hepacivirus/immunology ; Hepatitis C, Chronic/*genetics/immunology ; Humans ; Killer Cells, Natural/immunology/virology ; Male ; Middle Aged ; Receptors, KIR/*genetics/immunology ; Republic of Korea ; T-Lymphocyte Subsets/immunology

OBJECTIVE: The aim of this study was to establish three-dimensionally cultured endometrial cell model containing endometrial stromal cell (ESC), endometrial epithelial cell (EEC) and extracellular matrix (ECM) and to compare the morphological and biomolecular expression patterns of this model with mid-luteal endometrium in vivo. MATERIALS AND METHODS: The EEC and ESC was obtained from hysterectomy specimen and cultured separately. The EEC was overlayered in Matrigel layer on ESC embedded in collagen. The model had been cultured for 48 h in DMEM medium containing estrogen and progesterone. The ultrastructure was evaluated by electron microscopy. The expression of integrins, cyclooxygenases and matrix metalloproteinases were examined by immunohistochemistry and zymography. RESULTS: EEC in three-dimensional culture model grew with polarity and tight junction and desmosome between cells were found. The formation of pinopodes was also detected. In three-dimensionally cultured endometrial cell model, the expression of integrin alpha1, alpha4, beta3, MMP-1, -2, -3 and 9 was detected which was not expressed in monolayer culture of EEC, ESC or ESC embedded in collagen. CONCLUSION: The three-dimensionally cultured endometrial cell model possessed the morphological and biomolecular characteristics of in vivo endometrium of implantation period. These characteristics could be achieved by paracrine interactions between ESC and EEC. This model may contribute to the studies of differentiation of endometrium, process of implantation and pathophysiology of implantation-related diseases.
Collagen ; Desmosomes ; Endometrium ; Epithelial Cells ; Estrogens ; European Union ; Extracellular Matrix ; Female ; Humans* ; Hysterectomy ; Immunohistochemistry ; Integrin alpha1 ; Integrins ; Matrix Metalloproteinases ; Microscopy, Electron ; Progesterone ; Prostaglandin-Endoperoxide Synthases ; Stromal Cells ; Tight Junctions

PURPOSE: We compared the oncologic outcomes of breast-conserving surgery plus radiation therapy (BCS+RT) and modified radical mastectomy (MRM) under anthracycline plus taxane-based (AT) regimens and investigated the role of adjuvant radiation therapy (RT) in patients with pathologic N1 (pN1) breast cancer treated by mastectomy. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 2,011 patients with pN1 breast cancer who underwent BCS+RT or MRM alone at 12 institutions between January 2006 and December 2010. Two-to-one propensity score matching was performed for balances in variables between the groups. RESULTS: The median follow-up duration for the total cohort was 69 months (range, 1 to 114 months). After propensity score matching, 1,074 patients (676 in the BCS+RT group and 398 in the MRM-alone group) were analyzed finally. The overall survival, disease-free survival, locoregional failure-free survival, and regional failure-free survival (RFFS) curves of the BCS+RT group vs. MRM-alone group were not significantly different. The subgroup analysis revealed that in the group with both lymphovascular invasion (LVI) and histologic grade (HG) III, the BCS+RT showed significantly superior RFFS (p=0.008). Lymphedema (p=0.007) and radiation pneumonitis (p=0.031) occurred more frequently in the BCS+RT group than in the MRM-alone group, significantly. CONCLUSION: There are no differences in oncologic outcomes between BCS+RT and MRM-alone groups under the AT chemotherapy regimens for pN1 breast cancer. However, BCS+RT group showed superior RFFS to MRM-alone group in the patients with LVI and HG III. Adjuvant RT might be considerable for pN1 breast cancer patients with LVI and HG III.
Anthracyclines ; Breast Neoplasms ; Breast ; Cohort Studies ; Disease-Free Survival ; Drug Therapy ; Follow-Up Studies ; Humans ; Lymphedema ; Mastectomy ; Mastectomy, Modified Radical ; Mastectomy, Segmental ; Medical Records ; Propensity Score ; Radiation Pneumonitis ; Retrospective Studies

OBJECTIVE: The aim of this study was to examine and compare the gastrointestinal (GI) risk factors and treatment patterns of rheumatoid arthritis (RA) and osteoarthritis (OA) patients in Korea. METHODS: This was a cross-sectional, observational study on RA and OA patients taking non-steroidal anti-inflammatory drugs (NSAIDs) for at least 1 month. A total of 1,896 patients (981 RA patients, 915 OA patients) were recruited from 20 university hospitals. Data were collected through medical records and patient surveys. GI risk factors included age, prolonged (over 3 months) or high-dose use of NSAIDs, alcohol drinking, smoking, use of aspirin, anticoagulants or glucocorticoids, comorbidities, and history of Helicobacter pylori infection or other GI complications. Treatment patterns were classified according to groups using, selective cyclooxygenase (COX)-2 inhibitors+/-gastro-protective agents, non-selective COX-2 inhibitors+proton pump inhibitor, or non-selective COX-2 inhibitors+/-other gastro-protective agents. RESULTS: GI risk factors were highly present in both RA and OA patients. The proportion of prolonged use of NSAIDs, smoking, and glucocorticoid use were higher in RA patients (p<0.001). The proportion of comorbidities and use of aspirin were higher in OA patients (p<0.001). The remaining GI risk factors were present in similar proportions in both groups. Use of selective COX-2 inhibitors or gastro-protective agents was higher in RA patients. CONCLUSION: Prolonged use of NSAIDs and concomitant glucocorticoid use were higher in RA patients, while comorbidities and concomitant aspirin use were predominant in OA patients. These results will provide insights for use in development of future guidelines for proper selection of NSAIDs and effective prevention of GI complications in arthritis patients.
Alcohol Drinking ; Anti-Inflammatory Agents, Non-Steroidal ; Anticoagulants ; Arthritis ; Arthritis, Rheumatoid* ; Aspirin ; Comorbidity ; Cyclooxygenase 2 Inhibitors ; Glucocorticoids ; Helicobacter pylori ; Hospitals, University ; Humans ; Korea* ; Medical Records ; Observational Study ; Osteoarthritis* ; Prostaglandin-Endoperoxide Synthases ; Risk Factors* ; Smoke ; Smoking