Background/Aims: Cigarette smoking is an important modifiable risk factor in kidney disease progression. However, the underlying mechanisms for this are lacking. This study aimed to assess whether nicotine (NIC), a major toxic component of cigarette smoking, would exacerbates tacrolimus (TAC)-induced renal injury. Methods: Sprague-Dawley rats were treated daily with NIC, TAC, or both drugs for 4 weeks. The influence of NIC on TAC-caused renal injury was examined via renal function, histopathology, oxidative stress, mitochondria, endoplasmic reticulum (ER) stress, and programmed cell death (apoptosis and autophagy). Results: Both NIC and TAC significantly impaired renal function and histopathology, while combined NIC and TAC treatment aggravated these parameters beyond the effects of either alone. Increased oxidative stress, ER stress, mitochondrial dysfunction, proinf lammatory and profibrotic cytokine expressions, and programmed cell death from either NIC or TAC were also aggravated by the two combined. Conclusions Our observations suggest that NIC exacerbates chronic TAC nephrotoxicity, implying that smoking cessation may be beneficial for transplant smokers taking TAC.

PURPOSE: Lapatinib is a candidate drug for treatment of trastuzumab-resistant, human epidermal growth factor receptor 2 (HER2)–positive gastric cancer (GC). Unfortunately, lapatinib resistance renders this drug ineffective. The present study investigated the implication of forkhead box O1 (FOXO1) signaling in the acquired lapatinib resistance in HER2-positive GC cells. MATERIALS AND METHODS: Lapatinib-resistant GC cell lines (SNU-216 LR2-8) were generated in vitro by chronic exposure of lapatinib-sensitive, HER2-positive SNU-216 cells to lapatinib. SNU-216 LR cells with FOXO1 overexpression were generated by stable transfection of a constitutively active FOXO1 mutant (FOXO1A3). HER2 and MET in SNU-216 LR cells were downregulated using RNA interference. The sensitivity of GC cells to lapatinib and/or cisplatin was determined by crystal violet assay. In addition, Western blot analysis, luciferase reporter assay and reverse transcription–polymerase chain reaction were performed. RESULTS: SNU-216 LR cells showed upregulations of HER2 and MET, but downregulation of FOXO1 compared to parental SNU-216 cells. FOXO1 overexpression in SNU-216 LR cells significantly suppressed resistance to lapatinib and/or cisplatin. In addition, FOXO1 negatively controlled HER2 and MET at the transcriptional level and was negatively controlled by these molecules at the post-transcriptional level. A positive crosstalk was shown between HER2 and MET, each of which increased resistance to lapatinib and/or cisplatin. CONCLUSION: FOXO1 serves as an important linker between HER2 and MET signaling pathways through negative crosstalks and is a key regulator of the acquired lapatinib resistance in HER2-positive GC cells. These findings provide a rationale for establishing a novel treatment strategy to overcome lapatinib resistance in a subtype of GC patients.
Blotting, Western ; Cell Line ; Cisplatin ; Down-Regulation ; Drug Resistance ; Gentian Violet ; Humans ; In Vitro Techniques ; Luciferases ; Parents ; Receptor, Epidermal Growth Factor ; Receptor, ErbB-2 ; RNA Interference ; Stomach Neoplasms ; Transfection ; Up-Regulation

PURPOSE: We previously reported that forkhead transcription factors of the O class 1 (FOXO1) expression in gastric cancer (GC) was associated with angiogenesis-related molecules. However, there is little experimental evidence for the direct role of FOXO1 in GC. In the present study, we investigated the effect of FOXO1 on the tumorigenesis and angiogenesis in GC and its relationship with SIRT1. MATERIALS AND METHODS: Stable GC cell lines (SNU-638 and SNU-601) infected with a lentivirus containing FOXO1 shRNA were established for animal studies as well as cell culture experiments. We used xenograft tumors in nude mice to evaluate the effect of FOXO1 silencing on tumor growth and angiogenesis. In addition, we examined the association between FOXO1 and SIRT1 by immunohistochemical tissue array analysis of 471 human GC specimens and Western blot analysis of xenografted tumor tissues. RESULTS: In cell culture, FOXO1 silencing enhanced hypoxia inducible factor-1alpha (HIF-1alpha) expression and GC cell growth under hypoxic conditions, but not under normoxic conditions. The xenograft study showed that FOXO1 downregulation enhanced tumor growth, microvessel areas, HIF-1alpha activation and vascular endothelial growth factor (VEGF) expression. In addition, inactivated FOXO1 expression was associated with SIRT1 expression in human GC tissues and xenograft tumor tissues. CONCLUSION: Our results indicate that FOXO1 inhibits GC growth and angiogenesis under hypoxic conditions via inactivation of the HIF-1alpha-VEGF pathway, possibly in association with SIRT1. Thus, development of treatment modalities aiming at this pathway might be useful for treating GC.
Angiogenesis Modulating Agents ; Animals ; Anoxia ; Blotting, Western ; Carcinogenesis ; Cell Culture Techniques ; Cell Line ; Down-Regulation ; Forkhead Transcription Factors ; Heterografts ; Humans ; Lentivirus ; Mice ; Mice, Nude ; Microvessels ; RNA, Small Interfering ; Stomach Neoplasms* ; Tissue Array Analysis ; Transcription Factors* ; Vascular Endothelial Growth Factor A

PURPOSE: We sought to evaluate the clinical usefulness of decision making by a multidisciplinary heart team for identifying potential candidates for transcatheter aortic valve implantation (TAVI) in patients with symptomatic severe aortic stenosis. MATERIALS AND METHODS: The multidisciplinary team consisted of two interventional cardiologists, two cardiovascular surgeons, one cardiac imaging specialist, and two cardiac anesthesiologists. RESULTS: Out of 60 patients who were screened as potential TAVI candidates, 31 patients were initially recommended as appropriate for TAVI, and 20 of these 31 eventually underwent TAVI. Twenty-two patients underwent surgical aortic valve replacement (AVR), and 17 patients received only medical treatment. Patients who underwent TAVI and medical therapy were older than those who underwent surgical AVR (p<0.001). The logistic Euroscore was significantly highest in the TAVI group and lowest in the surgical AVR group (p=0.012). Most patients in the TAVI group (90%) and the surgical AVR group (91%) had severe cardiac symptoms, but only 47% in the medical therapy group had severe symptoms. The cumulative percentages of survival without re-hospitalization or all-cause death at 6 months for the surgical AVR, TAVI, and medical therapy groups were 84%, 75%, and 28%, respectively (p=0.007, by log-rank). CONCLUSION: TAVI was recommended in half of the potential candidates following a multidisciplinary team approach and was eventually performed in one-third of these patients. One-third of the patients who were initially considered potential candidates received surgical AVR with favorable clinical outcomes.
Aged, 80 and over ; Aortic Valve Stenosis/*surgery ; Decision Making ; Female ; Humans ; Male ; *Patient Care Team ; Postoperative Complications/epidemiology ; Risk Factors ; Severity of Illness Index ; *Transcatheter Aortic Valve Replacement ; Treatment Outcome

PURPOSE: Transcatheter aortic valve implantation (TAVI) has become an attractive therapeutic strategy for severe aortic stenosis (AS) in elderly patients due to its minimally-invasive nature. Therefore, early results of its clinical outcomes in elderly Korean patients were evaluated. MATERIALS AND METHODS: We compared early clinical outcomes of TAVI, surgical aortic valve replacement (SAVR), and optimal medical therapy (OMT) in patients aged > or =80 years with symptomatic severe AS. Treatment groups were allocated as follows: TAVI (n=10), SAVR (n=14), and OMT (n=42). RESULTS: Baseline clinical characteristics including predicted operative mortality were similar among the three groups. However, patients with New York Heart Association functional class III or IV symptoms and smaller aortic valve area were treated with TAVI or SAVR rather than OMT. In-hospital combined safety endpoints (all-cause mortality, major stroke, peri-procedural myocardial infarction, life-threatening bleeding, major vascular complication, and acute kidney injury) after TAVI or SAVR were significantly lower in the TAVI group than in the SAVR group (10.0% vs. 71.4%, respectively, p=0.005), along with an acceptable rate of symptom improvement and device success. During the follow-up period, the TAVI group showed the lowest rate of 3-month major adverse cardiovascular and cerebrovascular events, a composite of all-cause mortality, myocardial infarction, major stroke, and re-hospitalization (TAVI 0.0% vs. SAVR 50.0% vs. OMT 42.9%, p=0.017). CONCLUSION: Treatment with TAVI was associated with lower event rates compared to SAVR or OMT. Therefore, TAVI may be considered as the first therapeutic strategy in selected patients aged > or =80 years with symptomatic severe AS.
Age Factors ; Aged, 80 and over ; Aortic Valve/*surgery ; Aortic Valve Stenosis/diagnosis/*surgery ; Female ; Humans ; Male ; Prognosis ; Risk Factors ; Treatment Outcome

BACKGROUND: Kidney transplantation (KT) is the optimal treatment for end stage renal disease. However, the relative shortage of organs for transplantation (from human leukocyte antigen- or ABO incompatible [ABOi] living donors) has led to ABOi KT as an accepted method to expand the pool of living kidney donors. To date, reports of the outcomes of ABOi KT are limited; therefore this study aims to evaluate the outcomes of ABOi KT in recipients. METHODS: We identified 45 patients who underwent live-donor ABOi KT between February 2007 and November 2011 at Maryknoll Medical Center. All of them were treated according to the scheduled protocol of plasmapheresis with low dose intravenous immunoglobulin, and low dose rituximab- or tacrolimus-based triple immunosuppressant regimens. Clinical parameters and the incidence of rejections in these patients were analyzed. RESULTS: We had three cases (6.6%) of biopsy-proven acute antibody-mediated rejections and one case (2.2%) of acute cellular rejection, all of which were successfully treated. The median follow-up duration was 20 months (range, 2~59). Antibody depletion was scheduled according to baseline anti-ABO antibody titer (tube method: median immunoglobulin G titer/immunoglobulin M titer 64 [range, 8~4,096]/16 [range, 2~256], respectively). Although there was no patient death, one patient lost his graft due to nonadherence to immunosuppressants. CONCLUSIONS: Our analysis of ABOi KT has shown excellent and promising outcomes. These practices may therefore represent an acceptable option for expanding the pool of living kidney donors.
Follow-Up Studies ; Humans ; Immunoglobulin G ; Immunoglobulins ; Immunosuppression ; Incidence ; Kidney ; Kidney Failure, Chronic ; Kidney Transplantation ; Leukocytes ; Plasmapheresis ; Rejection (Psychology) ; Tissue Donors ; Transplants

BACKGROUND: Ginseng has been used to manage various types of pain in folk medicine. This study characterized the effect of treatment with intrathecal ginsenosides, the active components of ginseng in a postoperative pain model. METHODS: Male Sprague-Dawley rats were implanted with lumbar intrathecal catheters. An incision was made in the plantar surface of the hindpaw. Withdrawal thresholds following the application of a von Frey filament to the wound site were measured. To determine the role of the opioid or GABA receptors following treatment with the ginsenosides, naloxone, bicuculline (a GABAA receptor antagonist), and saclofen (a GABAB receptor antagonist) were administered intrathecally 10 min before the delivery of the ginsenosides and the changes of the withdrawal thresholds after application of the von Frey filament were observed. RESULTS: Treatment with the intrathecal ginsenosides increased the withdrawal threshold in a dose dependent manner. Pre-treatment with intrathecal naloxone reversed the antinociceptive effect of the ginsenosides. However, pre-treatment with intrathecal bicuculline and saclofen failed to have an effect on the activity of the ginsenosides. CONCLUSIONS: These results suggest that ginsenosides are effective to alleviate the postoperative pain evoked by paw incision. The opioid receptor, but not GABA receptors, may be involved in the antinociceptive action of the ginsenosides at the spinal level.
Analgesia ; Animals ; Bicuculline ; Catheters ; Ginsenosides* ; Humans ; Male ; Medicine, Traditional ; Models, Animal* ; Naloxone ; Pain, Postoperative* ; Panax ; Rats* ; Rats, Sprague-Dawley ; Receptors, GABA ; Receptors, Opioid ; Spinal Cord ; Wounds and Injuries

BACKGROUND: Ischemia-reperfusion injury related to unsuccessful myocardial protection affects postoperative ventricular function and mortality during open-heart surgery. We prospectively compared the effects of administration of histidine-tryptophan-ketoglutarate (HTK) solution and cold blood cardioplegia (CBC) on myocardial protection and clinical outcome in patients undergoing mitral valve surgery. MATERIAL AND METHOD: Seventy patients with mitral regurgitation (MR) undergoing mitral valve surgery were randomly divided into the HTK group (n=31) and the CBC group (n=31); eight patients were excluded. Perioperative hemodynamics, cardiac medications, pacing, postoperative outcomes and complications were recorded during the hospital stay. All patients received follow-up for at least 6 months postoperatively for morbidity and mortality. RESULT: There were no significant differences in the hemodynamics between the groups during the study period, except for the mean pulmonary artery pressure (MPAP), PCWP and CVP that were lower in the HTK group at 15 min after weaning of CBP. There were no differences for inotropic support and pacing during the 12 hrs postoperatively between the groups. CK-MB values on day 1 and day 2 were 77+/-54 and 41+/-23 for the HTK group and 70+/-69 and 44+/-34 for the CBC group, respectively (p=NS). Postoperative clinical outcomes were similar in both groups for at least 6 months during the follow-up period. CONCLUSION: These results suggest that the use of HTK solution is as safe as cold blood cardioplegia in terms of myocardial protection.
Cardioplegic Solutions* ; Follow-Up Studies ; Heart Arrest, Induced ; Hemodynamics ; Humans ; Length of Stay ; Mitral Valve Insufficiency ; Mitral Valve* ; Mortality ; Prospective Studies ; Pulmonary Artery ; Reperfusion Injury ; Ventricular Function ; Weaning

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)induces apoptosis in some cancer cells such as breast,prostate,lung,and colon cancer cells,but not normal cells.However,because the effects of TRAIL in gastric cancer cells is unclear,we undertook this study to clarify the effects of TRAIL and its mechanism. To assess the cytotoxicity of TRAIL,two human gastric cancer cell lines,SNU-484 and SNU601,were treated with TRAIL (0-200 ng/mL)in the presence or absence of cycloheximide (1 microgram/mL)for 24 h.Both SNU-484 and SNU-601 were sensitive to TRAIL-induced cell death in a dose-dependent manner.The combination of TRAIL (100 ng/mL)and cycloheximide (1 microgram/mL)for 24 h enhanced cell death and PARP cleavage by promoting activations of caspase-8, caspase-9,and caspase-3,relative to that of TRAIL alone.We further examined the expressions of death receptor 4 (DR4),death receptor 5 (DR5),and FLICE inhibitory protein (FLIP).Although DR4 and DR5 were expressed in both cell lines,the expression of long form (FLIPL )and short form (FLIPS )of FLIP were detected at the low levels. Overexpression of FLIPL or FLIPS in both cell lines rendered the cells resistant to TRAIL.Taken together,our results suggest that FLIP promotes human gastric cancer cell survival against TRAIL-induced apoptosis and is important modulator for TRAIL-induced cell death in human gastric cancer cells.
Apoptosis ; CASP8 and FADD-Like Apoptosis Regulating Protein ; Caspase 8 ; Cell Death ; Cell Line ; Cell Survival ; Colonic Neoplasms ; Cycloheximide ; Humans* ; Necrosis* ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; Stomach Neoplasms*

Although protein kinase B (PKB, AKT) has been investigated extensively for its roles in oncogenic transformation and apoptotic prevention, controversial results are also reported. Here we assessed the role of AKT in the cell growth and expression of a key set of cell cycle regulators in the human normal uroepithelial cell line, SVHUC-1. AKT activity was suppressed by permanent transfection of dominent negative (DN)-AKT with Lipofectamine Plus. Cell viability was measured by the crystal violet assay. DNA contents stained by propidium iodide were measured by flow-cytometry for cell cycle analysis. Cell growth curve showed that overexpression of DN-AKT which suppressed the AKT activity decreased the cell growth. In the cell cycle analysis, overexpression of DN-AKT resulted in a 6% increase in the proportion of cells in G1 phase. Furthermore, DN-AKT overexpression increased the p27(Kip1) protein expression and the activation of a transcription factor FKHR, which induces p27(Kip1) transcription. Our results suggest that, in normal uroepithelial cells, AKT activation increases the cell growth through the influence on p27(Kip1) expression and FKHR activation. Thus, AKT may be used as a biomarker for tumor transformation of bladder uroepithelial cells.
Cell Cycle ; Cell Line* ; Cell Survival ; DNA ; G1 Phase ; Gentian Violet ; Humans ; Propidium ; Proto-Oncogene Proteins c-akt ; Transcription Factors ; Transfection ; Urinary Bladder