OBJECTIVES: Cadherin/catenin adhesion complex is fundamentally involved in epithelial cancer invasion and metastasis. E-cadherin and EGFR colocalize on the basolateral membrane of epithelial cell and EGF down-regulate E-cadherin expression. In the invasion and metastasis of cancer, E-cadherin expression is decreased and growth factors receptor is overexpressed. The present study was aimed to find the role of E-cadherin, beta-and gamma-catenin, growth factors and its receptors in cervical cancer cell lines. METHODS: The cervical cancer cell cultures were treated with different time duration of EGF 30 ng/ml and TGF-a 10 ng/ml(0, 10 min, 20 min, 30 min, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr). The change in cancer cell morphology and the changes in E-cadherin, beta- and gamma-catenin, EGFR and activated EGFR expression were studied with a western blot analysis and an immunoprecipitation. RESULTS: Through a western blot analysis, E-cadherin 120 kDa band and EGFR 170 kDa band were expressed in CaSki, HT-3 and ME-180 cell line, which showed epithelial contact growth. 1n these 3 cell lines, expression of E-cadherin did not decrease with time dependent manner. after the treatment of EGF and TGF- alpha. The expression of EGFR decreased and activated EGFR expression increased in 30 minutes to 1 hour but decreased subsequently. When the cells treated with EGF, there were no change in beta-and gamma-catenin expression with there dependent manner. The tyrosine phosphorylation of beta-and gamma-catenin increased in 30 minutes to 1 hour but decreased subsequently with activated EGFR. CONCLUSION: This study showed that an activated EGFR which has involved with tyrosine phosphorylation of beta- and gamma-catenin influenced by growth factors rather than expression of E-cadherin, has a role in the invasion and metastasis of the cervical cancer.
Blotting, Western ; Cadherins* ; Cell Culture Techniques ; Cell Line* ; Epidermal Growth Factor* ; Epithelial Cells ; gamma Catenin* ; Immunoprecipitation ; Intercellular Signaling Peptides and Proteins ; Membranes ; Neoplasm Metastasis ; Phosphorylation* ; Transforming Growth Factor alpha* ; Tyrosine* ; Uterine Cervical Neoplasms*

OBJECTIVE: Neonatal blood pressure is one of the vital signs used to assess the general health of a newborn infant. But, it is difficult to measure and varies according to the model of sphygmomanometer, the used method, the postconceptional age, birth weight, asphyxia, postnatal age and activity. We have assessed the rnean value of blood pressure(BP) according to postconceptional age. Variations caused by the postconceptional age, birth weight, body surface area(BSA), postnatal age, and sex were analyzed to determine the rnost important factor influencing the neonatal BP. METHODS: This study included 448 normal fullterm infants and healthy preterm infants born from Jan. 1995 to Aug. 1997 at Ewha Dongdaemoon and Mokdong Hospital. The BP was taken on day 1, 3, 5 with the oscillometric device(Dinamap 1846 critikon Inc.), for premature infants BP was followed up every week afterward. RESULTS: The neonatal BP was related to the postconceptional age(R=0.204, P<0.05), BSA(R =0.191, P<0.05), birth weight(R=0.183, P<0.05) and the postnatal age in fullterm infants(R=0.022, P<0.05) but not with the sex of the infant. The postconceptional age was the most significant variable among those influencing the BP. The normal value of the systolic and the diastolic BP was expressed as mean+/-2SD according to the postconceptional age, less than 28 weeks(n=27) systolic/diastolic pressure 50.4+/-23.1mmHg/26.2+/-12.8mmHg, 28-32weeks(n= 63) 60.1+/- 30.6mmHg/33.6+/-16.7mmHg, 32-36weeks(n = 139) 63.2+/- 20.4mmHg/37.7+/- 19.8 mmHg, 36- 40weeks(n=543) 65.6+/-18.5mmHg/38.6+/-17.3mmHg, more than 40weeks(n= 136) 67.7+/-16,9mmHg/39.3+/- 15.8mmHg. The increase in blood prssure with the postconceptional age was statistically significant. CONCLUSION: The normal range of neonatal BP, especially preterm infants, was presented, and it would be much valuable for the evaluation of BP in neonate if standardized by postconceptional age.
Asphyxia ; Birth Weight ; Blood Pressure* ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Parturition ; Reference Values* ; Sphygmomanometers ; Vital Signs

No abstract available.
Animals ; Capsaicin* ; Ganglia, Spinal* ; Rats*

No abstract available.
Animals ; Capsaicin* ; Ganglia, Spinal* ; Rats* ; Spinal Nerve Roots*

The expression of c-fos and c-jun in the brain of the rat after capsaicin treatment was investigated by in situ hybridization, dot blot hybridization and immunocytochemical methods. Adult male Sprague-Dawley rats[200g] were used for this study. The first set of rats received a single subcutaneous injection of capsaicin[50mg/Kg] dissolved in 10% Tween-80 and 10% ethanol in saline. The rats were decapitated 1, 3, 5, 10, 24, 72 hours and 1 week after capsaicin treatment. The control set of rats were treated with saline instead of capsaicin. In situ hybridization and dot blot hybridization were carried out. O1igonucleotide probe complimentary to c-fos mRNA sequences were used for this study and labeling of oligonucleotides was accomplished using the DNA tailing kit. The expression of c-fos mRNA on the nucleus of neurons in in situ hybridization was observed throughout the brain, and was especially abundant in the olfactory cortex, nucleus of diagonal band of Broca, habenular nuclei, periaqueductal gray, parabrachial nucleus, entopeduncular nucleus, ventral posterolateral nucleus of the thalamus and cerebellum. Compared to the control rats, c-fos mRNA were increased 24 hours after capsaicin injection and gradually decreased after 72 hours, returning to the normal control level 1 week after capsaicin injection. c-fos mRNA was detected only 1 week after capsaicin injection in the various areas of the brain. The fos protein-like immunoreactivity was initially somewhat decreased at 24 hours, but increased at 72 hours and reactions was maximally observed at 1 week after capsaicin treatment. But Jun protein immunoreactivity was not increased, on the contrary, it was even decreased both in numbers of reactive cells and immunoreactivity 1 week after capsaicin injection. From the above results, c-fos gene expression was pronounced in the nucleus concerned with pain, olfaction and taste such as VPL nucleus of the thalamus, olfactory cortex and parabrachial nucleus, in the limbic system concerned with stress and emotion such as nucleus of diagonal band of Broca, periaqueductal gray and habenular nucleus, in the structure concerned with somatic motor function such as entopeduncular nucleus and cerebellum. Also, the c-fos gene was activated by the capsaicin early in the course of effects, then the fos protein increased as a results of c-fos activation. On the other hand, c-jun did not respond to capsaicin treatment early in the course, but Jun protein decreased late in the course of capsaicin effects.
Adult ; Animals ; Brain* ; Capsaicin* ; Cerebellum ; DNA ; Entopeduncular Nucleus ; Ethanol ; Genes, fos ; Habenula ; Hand ; Humans ; In Situ Hybridization ; Injections, Subcutaneous ; Limbic System ; Male ; Neurons ; Olfactory Pathways ; Oligonucleotides ; Periaqueductal Gray ; Rats* ; Rats, Sprague-Dawley ; RNA, Messenger ; Septal Nuclei ; Smell ; Thalamus ; Ventral Thalamic Nuclei

Recently, it has been postulated that diabetic autonomic neuropathy is caused by reduction in availability of nerve growth factor (NGF) in enteric nervous system. This experiments were performed to determine the changes of the distribution of enteric neuropeptide by diabetes and these changes could be prevented by administration of NGF. Sprague Dawley rats (200~250gm) were made diabetic by a single intraperitoneal injection of streptozotocin 65 mg/kg in saline. Recombinant human NGF (Sigma, Co., Ltd.) were administered at a dose of 500ng/kg subcutaneously every day for consecutive 4 weeks after streptozotocin administration. After 4 weeks, rats were anesthetized with ether and perfused with 4% paraformaldehyde. ileum was dissected and prepared by whole mount preparation method. Prepared segments were immunostained for substance p, calcitonin gene-related peptide, vasoactive intestinal peptide, and galanin by PAP technique. For the observation of the interstitial cells of Cajal, segments were immersed in Champy-Maillet solution for 2 days Results obtained were as follows: 1. In myenteric plexus of diabetic rats, substance P-like and VIP-like immunoreactivity were not changed compared with that of the control group. CGRP-like and galanin-like immunoreactivity were decreased in diabetic group and immunoreactive cells for CGRP and galanin were also decreased 18.1% (P<0.01) and 43.7% (P<0.01) respectively. 2. In NGF administerd diabetic group, immunoreactivity of substance p, VIP, galanin in myenteric plexus were slightly increased and immunoreactive cells for substancre p, VIP, galanin were almost the same as that of the control group. However, immunoreactive cells for CGRP of myenteric plexus were not changed by NGF. 3. In submucous plexus of diabetic rats, immunoreactivity of all four neuropeptides(substance p, CGRP, VIP, galanin) were decreased compared with that of the control group. Immunoreactive cells for substance p, CGRP, VIP, and galanin were also decreased in 38.8%, 77.6%, 33.0%, and 35.7%, respectively (P<0.01). 4. In NGF administered diabetic group, immunoreactivities of substance p, VIP and galanin in submucous plexus were increased and the immunoreactive cells were increased significantly compared to diabetic group. However, immunoreactive cells for CGRP of submucous plexus were not changed by NGF. 5. Interstitial cells of Cajal of diabetic group were decreased 7.4% ovoidal cells (A type) and 28.3% round cells (B type) In NGF administered group, the morphology and the number of ICC were not different to the control group. With the above results, it could be assumed that NGF prevent the damage of neurotransmitter and ICC in enteric nervous system.
Animals ; Calcitonin Gene-Related Peptide ; Diabetic Neuropathies ; Enteric Nervous System ; Ether ; Galanin ; Humans ; Ileum ; Injections, Intraperitoneal ; Interstitial Cells of Cajal ; Myenteric Plexus ; Nerve Growth Factor* ; Neuropeptides ; Neurotransmitter Agents ; Peristalsis* ; Rats* ; Rats, Sprague-Dawley ; Streptozocin ; Submucous Plexus ; Substance P ; Vasoactive Intestinal Peptide

OBJECTIVE: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. METHODS: Mouse ovaries were fixed and cut into serial sections (7 micrometer thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell IITM system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase -polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). RESULTS: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GAPDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 microl with 50 oocytes, thus the resting 19.75 microl cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. CONCLUSION: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.
Alcohols ; Animals ; DNA, Complementary ; Eosine Yellowish-(YS) ; Ethanol ; Female ; Frozen Sections ; Gene Expression* ; Genes, Essential ; Guanidine ; Hematoxylin ; Mice ; Microdissection* ; Oocytes* ; Ovary ; Paraffin ; Polymerase Chain Reaction ; Ribonucleases ; RNA Stability ; RNA* ; RNA, Messenger ; RNA-Directed DNA Polymerase ; Xylenes

End-stage renal disease(ESRD) is the fatal retention of non-volatile, metabolic waste products, salt, and water due to extensive loss of functioning nephrons. Renal replacement therapy is primarily aimed to remove retained waste products and fluid. Adequacy of dialysis is the dose of dialysis below which one observes a significant worsening of morbidity and mortality. Urea kinetic modeling, Kt/Vurea, etc. is regarded as a quantitative guideline of dialysis adequacy in both hemodialysis(HD) and peritoneal dialysis(PD). Water is one of the most important uremic toxin retained in ESRD patients. The importance of fluid overload on morbidity and/or mortality in dialysis patients is yet to be evaluated. Recent technology of HD provides adequate Kt/ Vurea for relatively short dialysis time and higher patient survival. Blood pressure control and extracellular fluid(ECF) volume are closely related with dialysis time in HD patients. Short dialysis time, 3-4 hours per session may not enough to control blood pressure. Hypertension is an important risk factor of survival in dialysis patients. Fluid overload is the most important factor of hypertension in dialysis patients. Patients with uncontrolled hypertension have higher mortality rate despite similar Kt/Vurea compared with patients with good blood pressure control. A longer dialysis time improves ECF volume and blood pressure control and decreases cardiovascular mortality. Continuous ambulatory peritoneal dialysis(CAPD) is a slow continuous therapy and is believed to maintain better control of ECF volume and blood pressure compared with hemodialysis. ECF volume and blood pressure controls are improved after initiation of CAPD, however, return to pre-CAPD levels after a few years of CAPD when residual renal function disappears. Patients transferred to hernodialysis from CAPD lose around 4kg of body weight for a few months on HD. Approximately 25% of CAPD patients are clinically fluid overloaded. Increased peritoneal permeability is an independent risk factor for patient and technique survivals in CAPD patients. Fluid overload in CAPD patients with increased peritoneal pernability is believed to be an important underlying mechanism of increased mortality and technique failure in this group. In conclusion, fluid overload is an important risk factor of mortality in dialysis patients. Sufficient dialysis time resulting good controls of ECF volume and blood pressure in hemodialysis patients and individualized dialysis prescription according to the peritoneal permeability in CAPD patients are vital to provide adequate dialysis and to decrease cardiovas-cular mortality.
Blood Pressure ; Body Weight ; Dialysis* ; Humans ; Hypertension ; Kidney Failure, Chronic ; Mortality ; Nephrons ; Peritoneal Dialysis, Continuous Ambulatory ; Permeability ; Prescriptions ; Renal Dialysis ; Renal Replacement Therapy ; Risk Factors ; Urea ; Waste Products ; Water

This year marks the 40th anniversary of the introduction of National Health Insurance (NHI) which has contributed to improving public health and accessibility. This article aims to show the trends of main indicators during the last 40 years. NHI has achieved rapid expansion of target population (1977–1989). The percentage of population covered increased from 8.8% in 1977 to 94% in 1990. The average number of visit days per person was 0.75 in 1977 but significantly increased to 31.11 in 2015. In 2015, NHI revenues were 52.4 trillion won and expenditures were 48.2 trillion won which is 9.5 times and 9.6 times higher than in 1995. NHI achieved universal coverage in short period of time and has contributed to improving the healthcare status. However, there still remain problems including low-benefit coverage and high out of pocket money. Therefore, the effort to reform these problems is needed.
Anniversaries and Special Events ; Delivery of Health Care ; Health Expenditures ; Health Services Needs and Demand ; Humans ; National Health Programs* ; Public Health ; Universal Coverage

This year marks the 40th anniversary of the introduction of National Health Insurance (NHI) which has contributed to improving public health and accessibility. This article aims to show the trends of main indicators during the last 40 years. NHI has achieved rapid expansion of target population (1977–1989). The percentage of population covered increased from 8.8% in 1977 to 94% in 1990. The average number of visit days per person was 0.75 in 1977 but significantly increased to 31.11 in 2015. In 2015, NHI revenues were 52.4 trillion won and expenditures were 48.2 trillion won which is 9.5 times and 9.6 times higher than in 1995. NHI achieved universal coverage in short period of time and has contributed to improving the healthcare status. However, there still remain problems including low-benefit coverage and high out of pocket money. Therefore, the effort to reform these problems is needed.
Anniversaries and Special Events ; Delivery of Health Care ; Health Expenditures ; Health Services Needs and Demand ; Humans ; National Health Programs* ; Public Health ; Universal Coverage