OBJECTIVE: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. MATERIAL AND METHODS: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and 5microgram/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal perm of hybrid F1 male mice(1x106/ml). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, blastocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identity ES cells, the surface markers alkaline phosphatase, SSEA-1, 3, 4 and Oct4 staining were examined in replated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. RESULTS: Although the cleavage rate (> or =2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic blastocysts (9.6+/-3.1, 35.1+/-5.2) were significantly lower than those of IVF blastocysts (19.5+/-4.7, 63.2+/-13.0) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-1 and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac differentiation derived from mES or P-mES cells was confirmed. CONCLUSION: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.
Alkaline Phosphatase ; Animals ; Antigens, CD15 ; Bisbenzimidazole ; Blastocyst ; Cell Count ; Cell Line ; Embryonic Stem Cells* ; Embryonic Structures ; Ethanol ; Female ; Fertilization in Vitro* ; Humans ; Male ; Mice* ; Oocytes ; Propidium

OBJECTIVE: This study was to investigate the generation of the functional neuron derived from human embryonic stem(hES,MB03) cells on in vitro neural cell differentiation system. METHODS: For neural progenitor cell formation derived from hES cells, we produced embryoid bodies (EB: for 5 days, without mitogen) from hES cells and then neurospheres (for 7~10 days, 20 ng/ml of bFGF added N2 medium) from EB. And then finally for the differentiation into mature neuron, neural progenitor cells were cultured in i) N2 medium only (without bFGF),ii) N2 supplemented with 20 ng/ml platelet derived growth factor-bb (PDGF-bb) or iii) N2 supplemented with 5 ng/ml brain derived neurotrophic factor (BDNF) for 2 weeks. Identification of neural cell differentiation was carried out by immunocytochemistry using betaIII-tubulin (1:100) and GFAP (1:500). Also, generation of functional neuron was identified using anti-glutamate (Sigma, 1:1000), anti-GABA (Sigma, 1:1000), anti-serotonin (Sigma, 1:1000) and anti-tyrosine hydroxylase (Sigma, 1:1000). RESULTS: In vitro neural cell differentiation, neurotrophic factors (PDGF and BDNF) treated cell groups were high expressed MAP-2 and GFAP than non-treated cell group. The highest expression pattern of MAP-2 and betaIII-tubulin was indicated in BDNF treated group. Also, in the presence of PDGF-bb or BDNF, mostof the neural cells derived from hES cells were differentiated into gultamate and GABA neuron in vitro. Furthermore, we confirmed that there were a few serotonin and tyrosine hydroxylase positive neuron in the same culture environment. CONCLUSION: This results suggested that the generation of functional neuron derived from hES cells was increased by addition of neurotrophic factors such as PDGF-bb or BDNF in b-FGF induced neural cell differentiation system and especially gultamate and GABA neurons were mainly produced in the system.
Blood Platelets ; Brain-Derived Neurotrophic Factor* ; Cell Differentiation* ; Embryoid Bodies ; Embryonic Stem Cells* ; gamma-Aminobutyric Acid ; Humans* ; Immunohistochemistry ; Nerve Growth Factors ; Neurons* ; Serotonin ; Stem Cells ; Tyrosine 3-Monooxygenase

No abstract available.
Acrosome* ; Fertilization* ; Humans* ; Male ; Spermatozoa*

This study was carried out to investigate the development rates of human embryos co-cultured with cumulus cells to each blastocyst stage. Human zygotes were co-cultured on cumulus cell monolayer in YS medium supplemented with 20% hFF. On day 2, if patient had four or more 'good' embryos(regular blastomeres without fragmentation), embryos were further cultured for 72hrs. Blastocysts on day 5 were classified into early blastocyst (ErB), early expanding blastocyst (EEB), middle expanding Blastocyst (MEB), and expanded blastocyst (EdB) on the basis of their morphological aspects of trophectoderm cells and blastocoele. Subsequently, maximum 3 of best blastocysts were transferred in 486 cycles. The results in this study were as follows: Patients who had four or more 'good' embryos on day 2 were 498 persons, but patients whose embryos could not be transferred due to failure in development to the blastocyst stage on day 5 were 12 persons (2.4%). The development rate of embryos to the blastocyst stage was 58.2% (2,885/4,957) on day 5, and the rates that developed to the ErB, EEB, MEB, and EdB stage were 15.0% (743/4,957), 14.9% (739/4,957), 14.4% (714/4,957), and 13.9% (689/4,957), respectively. Total 1366 blastocysts were transferred in 486 cycles (mean number=2.81). The implantation rate and the ongoing implantation rate obtained by observing the number of G-sac and FHB were 29.9% (409/1,366) and 22.5% (308/1,366), respectively. The clinical pregnancy rate was 51.2% (249/486), and the ongoing pregnancy rate was 39.1% (190/486). Among women showing ongoing pregnancy, women with singleton were 50% (95/190), women with twin were 37.9% (72/190), and women with triplet were 12.1% (23/190). Although triplet pregnancy rate in this study was high such as 12.1%, because many blastocysts with high viability were produced in our co-culture system using cumulus cells on day 5, we really believe that a multiple pregnancy except twin should not occur by selecting good embryos for maximum two blastocyst transfer. These results demonstrate that autologous cumulus cells may be used for the production of blastocysts with high developmental competence, and the use of autologous cumulus cells to be collected easily, and to be treated conveniently at OPU must be an effective means for obtaining high implantation and pregnancy rate.
Blastocyst* ; Blastomeres ; Coculture Techniques* ; Cumulus Cells* ; Embryo Transfer ; Embryonic Structures* ; Female ; Humans* ; Mental Competency ; Pregnancy ; Pregnancy Rate ; Pregnancy, Multiple ; Pregnancy, Triplet ; Triplets ; Zygote

BACKGROUND: The aim of this study was to examine the efficacies of somatosensory evoked potential (SEP) and diffusion-weighted magnetic resonance imaging (DWI) in predicting the clinical prognosis of comatose patients following cardiac arrest. METHODS: Forty-one patients resuscitated from out-of hospital cardiac arrest (OHCA) were retrospectively studied. After return of spontaneous circulation (ROSC), SEP was conducted between one and three days after resuscitation, and DWI was conducted within five days of resuscitation. SEP was classified into three grades: normal, delayed conduction or unilateral loss of the N20 peak, and bilateral loss of the N20 peak. Bilateral loss of the N20 peak was considered a predictor of poor prognosis. For DWI, diffuse signal intensity (SI) abnormality in the cerebral cortex or abnormality in other brain areas in addition to the bilateral cerebral cortex was taken as a predictor of poor prognosis. For patient clinical prognosis, the Glasgow-Pittsburgh Cerebral Performance Category (CPC) was used to evaluate neurological results at the time of discharge. Resulting CPC scores of 1 and 2 were considered as a favorable prognosis, and scores of 3, 4, and 5 were considered as a poor prognosis. Sensitivity, specificity, positive predictive value, and negative predictive value for the prediction of poor prognosis were analyzed for each test individually and for the combination of the two tests. RESULTS: Among the 41 subject patients, 31 underwent SEP, 30 underwent DWI, and 20 underwent both tests. The prognosis predictor of SEP (bilateral loss of the N20 peak) predicted poor prognosis with 56.5% sensitivity, 100% specificity, 100% positive predictive value, and 44.4% negative predictive value. The prognosis predictor of DWI (diffuse SI abnormality in the cerebral cortex or abnormality in other brain areas in addition to the bilateral cerebral cortex) predicted poor prognosis with 85% sensitivity, 100% specificity, 100% predictive value, and 76.9% predictive value. For patients who underwent both tests, the sensitivity and negative predictive value for the prediction of poor prognosis increased to 92.3% and 87.5%, respectively, and the specificity and positive predictive value were maintained at 100%. CONCLUSIONS: The accuracy of poor prognosis prediction for patients in prolonged comas after resuscitation is enhanced by combining the results of SEP and DWI along with the individual results of each test.
Brain ; Cerebral Cortex ; Coma* ; Dinucleoside Phosphates ; Evoked Potentials, Somatosensory* ; Heart Arrest* ; Humans ; Magnetic Resonance Imaging* ; Prognosis* ; Resuscitation ; Retrospective Studies ; Sensitivity and Specificity

All-trans retinoic acid (ATRA) is a highly effective treatment for acute promyelocytic leukemia (APL), a cytogenetically distinct subtype of acute myeloid leukemia (AML). However, ATRA-based treatment is not effective in other subtypes of AML. In non-APL AML, ATRA signaling pathway is impaired or downmodulated, and consequently fails to respond to pharmacological doses of ATRA. Therefore, complementary treatment strategies are needed to improve ATRA responsiveness in non-APL AML. In this study, we investigated the combined effect of ATRA and bromodomain inhibitor JQ1, proven to have potent anti-cancer activity mainly through inhibition of c-Myc. We showed that the combination of ATRA with JQ1 synergistically inhibited proliferation of AML cells. The synergistic growth inhibition was resulted from differentiation or apoptosis depending on the kind of AML cells. Concomitantly, the combined treatment of ATRA and JQ1 caused greater depletion of c-Myc and hTERT expression than each agent alone in AML cells. Taken together, these findings support the rationale for the use of the combination of ATRA and JQ1 as a therapeutic strategy for the treatment of AML.
Apoptosis ; Leukemia ; Leukemia, Myeloid, Acute ; Leukemia, Promyelocytic, Acute ; Tretinoin

BACKGROUND: Infectious complications commonly occur in the survivors of out-of-hospital cardiac arrest. The aim of our study was to describe the incidence, associated factors and outcome of infectious complications of the survivors of out-of-hospital cardiac arrest. METHODS: We conducted a retrospective analysis of 75 patients who survived out-of-hospital cardiac arrest. We collected the data on the demographics, the modes of cardiac arrest, the duration of CPR, the dose of epinephrine, the use of hypothermia, new infections, the duration of mechanical ventilation, the length of stay in the intensive care unit (ICU), recovery of consciousness and the mortality. RESULTS: New infections developed in 46.7% of the patients. Asystole was the most common rhythm (70.7%). The most common infectious complication was pneumonia (40.0%) urinary tract infection developed in 10 cases, vascular catheter local infection developed in 6 cases, primary blood stream infection developed in 3 cases, wound infection developed in 2 cases and pseudomembranous colitis developed in 1 case. The most common pathogens of pneumonia were Pseudomonas aeruginosa and Staphylococcus aureus. Blood cultures were obtained in 36 patients during the first 24 hr and the pathogen was isolated in three. The patients with infection had a longer duration of mechanical ventilation and a longer stay in the ICU (p < 0.001, p = 0.001). CONCLUSIONS: Infectious complications are common in survivors of out-of-hospital cardiac arrest and these infections are associated with a longer duration of mechanical ventilation and a longer stay in the ICU. The most common infectious complication was pneumonia and the pathogens of pneumonia were Pseudomonas aeruginosa and Staphylococcus aureus.
Cardiopulmonary Resuscitation ; Consciousness ; Demography ; Enterocolitis, Pseudomembranous ; Epinephrine ; Heart Arrest ; Humans ; Hypothermia ; Incidence ; Intensive Care Units ; Length of Stay ; Out-of-Hospital Cardiac Arrest ; Pneumonia ; Pseudomonas aeruginosa ; Respiration, Artificial ; Retrospective Studies ; Rivers ; Staphylococcus ; Staphylococcus aureus ; Survivors ; Urinary Tract Infections ; Vascular Access Devices ; Wound Infection

PURPOSE: This study was conducted to see the effect of alcohol ingestion on clinical features of acute drug intoxicated patients. METHODS: We prospectively investigated drug intoxicated patients who visited the emergency department 6 hours after acute poisoning from January 2004 to December 2007. Patients were classified into two groups according to serum alcohol levels: an alcohol group (serum alcohol level>10 mg/dl) and a non-alcohol group. The type of toxic material, age, sex, duration of time to arrive to the emergency department (ED) after poisoning, mean arterial pressure, respiratory rate, base excess level, AST level, serum creatinine level, cause of poisoning, suicide attempt, past psychiatric history, discharge against medical advice rate, and admission rate were checked. The initial and final Poisoning Severity Score (PSS), the Glasgow coma scale, the length of stay in the intensive care unit (ICU), the usage of a mechanical ventilator, and death rate were also checked. RESULTS: The study enrolled 222 intoxicated patients of which 75 fell into the non-alcohol group and 147 into the alcohol group. Alcohol ingestion of acute poisoning in males was higher than in females. The AST level and discharge against medical advice rates in the alcohol group were higher than the non-alcohol group. The base excess level, length of stay in ICU, past psychiatric history rate, and admission rates in the non-alcohol group were higher than the alcohol group. The PSS were not correlated with alcohol consumption between the two groups. CONCLUSION: Alcohol ingestion is not associated with PSS. However, alcohol ingestion is commonly found in acute drug intoxicated patients. The discharge against medical advice rate in the alcohol group was higher than the non-alcohol group.
Alcohol Drinking ; Arterial Pressure ; Creatinine ; Eating ; Emergencies ; Female ; Glasgow Coma Scale ; Humans ; Intensive Care Units ; Length of Stay ; Male ; Prospective Studies ; Respiratory Rate ; Severity of Illness Index ; Suicide ; Ventilators, Mechanical

PURPOSE: The purpose of this study was to assess the incidence, survival rate and risk factors of acute renal failure (ARF) in patients following cardiac arrest. METHODS: This was a retrospective analysis of 132 cases of comatose survival after cardiac arrest and these patients had received cardiopulmonary resuscitation (CPR) in a tertiary emergency department from April 1997 to May 2007. We analyzed the basic data and clinical data, and we compared the patients who had developed ARF (ARF group) following CPR with those who had not (the non-ARF group). RESULTS: Out of the 132 comatose survivors, ARF occurred in 54 patients (40.9%). The survival rate was higher in the non-ARF group (52.6%) than that in the the ARF group (16.7%) (p<0.0001). Both baseline renal insufficiency (OR 5.721, 95% CI 2.42-13.51; p<0.0001) and a peak creatine kinase level > 5000 IU/L (OR 8.527, 95% CI 1.78-40.91; p=0.043) were independent predictors of ARF. However, a history of coronary artery disease was not independently associated with renal failure, although it was more frequently found in patients with ARF (the ARF group). CONCLUSION: ARF occurred commonly in the post-resuscitation period. The survival rate was higher in the non-ARF group than that in the ARF group. The major factors contributing to ARF were both baseline renal insufficiency and a peak creatine kinase level > 5000 IU/L.
Acute Kidney Injury ; Cardiopulmonary Resuscitation ; Coma ; Coronary Artery Disease ; Creatine Kinase ; Emergencies ; Heart Arrest ; Humans ; Incidence ; Renal Insufficiency ; Retrospective Studies ; Risk Factors ; Survival Rate ; Survivors

PURPOSE: To identify a useful screening test leading to diagnosis of hypertension in the emergency department (ED). METHODS: This was a retrospective medical record review of adult patients (18> or =years of age) admitted to the ED at a tertiary care educational hospital, between January 1, 2010 and February 28, 2010. Only those patients with a triage systolic blood pressure greater than or equal to 140 mmHg, or a diastolic blood pressure greater than or equal to 90 mmHg, were enrolled. Data including baseline characteristics, basic metabolic panel (BMP), urinalysis, electrocardiogram (ECG), chest radiograph, and whether or not they were diagnosed with hypertension, were obtained. Multivariate analysis was performed to determine an appropriate screening test for diagnosis of hypertension. RESULTS: Of the 447 enrolled patients, 81(18.1%) were diagnosed with hypertension. Age above 35 years (Odds ratio [OR]=8.263; 95% Confidence interval (CI), 1.034-66.062; p=0.046), diagnosis of diabetes mellitus (DM) (OR=3.99; 95% CI, 1.582-10.064; p=0.003), left ventricular hypertrophy (LVH) (OR=4.348; 95% CI, 1.968-9.607; p<0.001), and suspected stage II hypertension (OR=2.699; 95% CI, 1.151-6.329; p=0.022) were independently associated with a positive hypertension diagnosis. The area under the Receiver operating characteristic (ROC) curve for a positive diagnosis of hypertension was 0.687(95% CI, 0.642-0.730). CONCLUSION: Age above 35 years, existence of DM or LVH, and suspected stage II hypertension may be useful data points for screening and diagnosis of hypertension in the ED.
Adult ; Blood Pressure ; Diabetes Mellitus ; Electrocardiography ; Emergencies ; Humans ; Hypertension ; Hypertrophy, Left Ventricular ; Mass Screening ; Medical Records ; Multivariate Analysis ; Retrospective Studies ; ROC Curve ; Tertiary Healthcare ; Thorax ; Triage ; Urinalysis