Animal models for human chronic pain syndromes were developed and widely used for pain research. One of thsese neuropathic pain model by Kim and Chung[1992] has many advantages for operation and pain elicitation. We have examined the c-fos protein, substance P, CGRP immunoreactivity in dorsal root ganglia and dorsal horn in this neuropathic model. About 50 Sprague-Dawley rats were used for this study. L5 and L6 spinal nerve were ligated tightly to produce neuropathic pain model. After 2, 4, 8, 16, 24 hours and 1 week of surgery, rats were anesthesized and sacrificed by perfusion through the left ventricle with saline followed by 0.1M phosphate buffer[pH 7.4] containing 3% paraformaldehyde, 3% glutaraldehyde, and 0.1% picric acid. After confirmation of the roots transected by the surgery, the L5 and L6 dorsal root ganglion and spinal cord were removed and processed for immunohistochemistry. All tissue sections were immunohistochemically stained for substance P, CGRP and c-fos by using the peroxidase-antiperoxidase[PAP] method. Count the number of immunostained substance P and CGRP dorsal root ganglion cells and c-fos immunoreactive dorsal horn cells and analyzed statistically with Mann-Whitney U test. The results are as follows. 1. The number of c-fos protein immunoreactive neurons in the superficial layer of dorsal horn were increased markedly at 2 hours after operation, gradually decreased to normal level 1 week after operation. 2. The number of c-fos protein immunoreactive neurons in the deep layer of dorsal horn were gradually increased to the peak 24 hours after operation, decreased to normal level 1 week after operation. 3. The number of substance P and CGRP immunoreactive L5 and L6 dorsal root ganglion neurons were decreased markedly at 1 week after pain model operation. In conclusion, after neuropathic pain model operation, c-fos protein were immediately expressed in the superficial layer of spinal dorsal horn, thereafter c-fos protein in the deep layer of spinal dorsal horn were expressed. CGRP and substance P immunoreactive neurons were decreased markedly 1 week after neuropathic pain model operation.
Animals ; Chronic Pain ; Ganglia, Spinal* ; Glutaral ; Heart Ventricles ; Horns* ; Humans ; Immunohistochemistry ; Models, Animal ; Neuralgia ; Neurons* ; Perfusion ; Peripheral Nervous System Diseases* ; Posterior Horn Cells ; Rats* ; Rats, Sprague-Dawley ; Spinal Cord ; Spinal Nerve Roots* ; Spinal Nerves ; Substance P

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

PURPOSE: Cortical dysplasia (CD) designates a diverse group of malformations resulting from one or more abnormalities in the development of the cerebral cortex. We investigated the findings of interictal SPECT and the diagnostic usefulness of interical and ictal SPECT according to pathological grading (PG) in comparison with MRI. MATERIALS AND METHODS: This study included 16 patients (M:F=9:7, age: 19.9+/-11.8 yrs) with pathologically proven CD. Tc-99m ECD SPECT was performed in all patients: interictal 11, interictal and ictal 3, ictal 2. MRI were obtained in all patients and image analysis was done blindly as to the result of SPECT. Pathologic findings of CD were classified into grade 1 (G1, dyslamination), grade 2 (G2, dysplastic neurons) and grade 3 (G3, balloon cells). We compared SPECT with MRI in lesions-to-lesions and analyzed the result according to PG. RESULTS: In SPECT and MRI, 38 and 27 lesions were visually recognized. In 14 interictal SPECT, variable findings in 35 lesions were demonstrated: 26 were hypoperfusion, 7 hyperperfusion, 2 heterotopic perfusion in the white matter. By comparison between two studies, missed lesions were founded: SPECT were 1 lesion, MRI 12. Review of missed 12 lesions of MRI were followed according to PG; G1 patients were 16.7% (4/19), G2 40.0% (6/15), and G3 50% (2/4). CONCLUSION: Interictal SPECT in CD showed variable findings such as hypoperfusion, hyperperfusion or heterotopic perfusion. However, for detection of missed CD on MRI, SPECT may help to detect a functional abnormality of the lesion with high PG.
Cerebral Cortex ; Humans ; Magnetic Resonance Imaging* ; Malformations of Cortical Development* ; Perfusion ; Tomography, Emission-Computed, Single-Photon*

PURPOSE: Food protein-induced enterocolitis syndrome (FPIES) is a symptom complex of vomiting and diarrhea caused by non-IgE mediated allergy to cow's milk and/or soy in young infants. Transforming growth factor (TGF)-beta has been reported to protect the epithelial barrier of the gut from foreign antigens. We studied the expression of type 1 and 2 TGF-beta receptors in the mucosa of small intestine to investigate their roles in the pathogenesis of FPIES. METHODS: Twenty-eight patients, aged 7 to 120 days (mean 49 days) who were diagnosed with FPIES by clinical criteria and challenge tests were included. Immunohistochemical stainings for type 1 and 2 TGF-beta receptors were performed on endoscopic duodenal biopsy specimens. RESULTS: Type 1 and 2 TGF-beta receptors were expressed in the villous and crypt epithelial cells but nearly absent in the lamina propria in both patients and controls. Type 1 TGF-beta receptor expression was significantly lower in the patients who had villous atrophy than in the patients who had not and in controls. The expression of type 1 TGF-beta receptor was negatively correlated with the severity of villous atrophy. Type 2 TGF-beta receptor expression showed no significant difference between the patients and controls. CONCLUSION: Our results suggests that the decreased activity of type 1 TGF-beta receptor is implicated in the pathogenesis of FPIES in young infants.
Atrophy ; Biopsy ; Diarrhea ; Enterocolitis* ; Epithelial Cells ; Humans ; Hypersensitivity ; Infant ; Intestine, Small ; Milk ; Mucous Membrane ; Receptors, Transforming Growth Factor beta ; Transforming Growth Factors ; Vomiting

BACKGROUND: As the average lifespan of human increases, osteoporosis and osteoporosis-related fractures have become major health care problems. Despite recent advances in medical treatment, few studies have assessed the recognition of osteoporosis in general adults. This study examined the recognition of osteoporosis and analysed the relating factors. METHODS: We made a survey in a rural area called Chunjunlee in Chunchon city. The questionnaire contained general characteristics of people, sociocultural factors, questions constructed to know the recognition of osteoporosis and factors that were presumed to influence people's recognition of osteoporosis. DEXA was performed on those who visited our medical office free of charge. RESULTS: The total number of people who responded to the questionnaire was 204. They were composed of 81 men and 123 women. Among the total, 83% of women and 72.8% of men said that they had heard about osteoporosis. There was no significant recognition score difference between men and women. In the items of recognition, wrong answer rate concerning the association between osteoporosis and musculoskeletal disorders such as osteoarthritis and low back pain was above 90%. The significant factors that influenced its recognition were age, education level, menopause state and newspaper/ magazine subscription. All men who were diagnosed with osteoporosis on DEXA initially said that they did not have osteoporosis. CONCLUSION: Many people had a conceptual confusion between osteoporosis and other muscular skeletal disorders such as osteoarthritis and low back pain. Physicians played no significant role in improvement of people's recognition of osteoporosis. Generally, men thought that osteoporosis was a problem in women only.
Adult ; Delivery of Health Care ; Education ; Female ; Gangwon-do ; Humans ; Low Back Pain ; Male ; Menopause ; Osteoarthritis ; Osteoporosis* ; Periodicals as Topic ; Periodicals

Pancreatic cancer patients often have a poor prognosis and suffer from nutritional problems. Malnutrition is characterized by weight loss and decreased dietary intake, and is common among pancreatic cancer patients. The objective of this report was to describe the changes in dietary intake, body weight, nutritional status, and metabolic rate on a continuum from the time of diagnosis until the end of life in a patient with pancreatic cancer. In summary, the patient's nutritional status gradually declined, accompanied by extreme weight loss and decreased dietary intake. Conversely, resting energy expenditure, measured by indirect calorimetry, increased from 24 kcal/kg/day to 35 kcal/kg/day. Nutritional management during cancer treatment is important but may be challenging in pancreatic cancer patients.
Basal Metabolism ; Body Weight* ; Calorimetry, Indirect ; Diagnosis ; Diet Records ; Energy Metabolism ; Humans ; Malnutrition ; Nutritional Status* ; Pancreatic Neoplasms* ; Prognosis ; Weight Loss

Overnight fasting before elective surgery has been the routine to reduce the risk of pulmonary aspiration. Recently, several international guidelines for preoperative fasting recommend to intake carbohydrate-containing fluids up to 2 to 3 hours before the induction of anesthesia to improve postoperative recovery. Based on the recommendations, we developed a "preoperative carbohydrate diet" provided for the preoperative patients. The purpose of this case report is to share our experience of applying preoperative carbohydrate loading prior to surgery.
Anesthesia ; Fasting ; Humans ; Insulin Resistance ; Pancreaticoduodenectomy*

This study was conducted to develop job standards for clinical dietitian administering clinical nutrition therapy to diabetic patients in hospitals. Based on DACUM (Developing A Curriculum) analysis of 17 members including clinical dietitians, professors majoring in clinical nutrition and researchers, information on duties, tasks and task elements of clinical dietitians for diabetes care were derived and applied to diabetes mellitus-specific clinical nutrition care in hospitals for evaluation. The final developed job standards for clinical dietitians for diabetes care included four duties, 19 tasks and 56 task elements. The duties consisted of nutrition assessment, nutrition diagnosis, nutrition intervention, and nutrition monitoring . evaluation. For application of diabetes mellitus-specific job standards in clinical nutrition care, 108 work activities were developed and classified into 90 basic and 18 recommended types. Performance rates of standardized jobs were 80.2% at nutrition assessment, 99.6% at nutrition diagnosis, 78.5% at nutrition intervention, and 32.9% at nutrition monitoring . evaluation. These results can be applied as guidelines to implement jobs for diabetes mellitus-specific clinical nutrition services in clinical settings. In addition, they would be useful for education standards in educational institutions for education and training of clinical dietitian.
Diabetes Mellitus ; Diagnosis ; Education ; Humans ; Job Description ; Nutrition Assessment ; Nutrition Therapy* ; Nutritionists*

PURPOSE: To investigate methods of isolating putative corneal epithelial stem cells from cultured limbal tissue. METHODS: Three extraction techniques were compared to identify an efficient method of obtaining a large number of viable corneal epithelial stem cells from the limbus. Limbal tissues were extracted by incubation at 37 degrees C or 4 degrees C for 1 or 16 hours, respectively, with 1.2U/ml dispase/trypsin or by treatment with 0.05% trypsin and 0.01% ethyldiaminetetraacetic acid (EDTA) at 37 degrees C in single procedure. Collected cells were cultured on NIH/3T3-seeded plates, and colony forming efficiency (CFE) was evaluated. Fluorescence activated cell sorting (FACS) was performed with a Coulter EPICS 753 after incubation with Hoechst 33342 and propidium iodide (PI). Hoechst negative cells were obtained using gates exhibiting low Hoechst blue with a 424/44 nm BP filter. Gated cells of each fraction were re-cultured to assess the capability of colony formation. RESULTS: The mean numbers of viable cells obtained from treatment with dispase and trypsin was 3x10(4) cell/ml and 8.06x10(5) cell/ml at 37 degrees C and 4 degrees C incubations; the number increased to 1.21x10(6) cell/ml with a trypsin/EDTA treatment (p<0.05). CFE was 9.67+/-2.13% and 6.63+/-2.35% in rabbit and human cells, respectively. Likewise, the Hoechst negative fraction was 3.61+/-0.42% and 5.21+/-4.91% in rabbit and human cells, respectively. The sorted Hoechst negative cells were cultured through four passages, forming small round colonies. In rabbit cells, the CFEs of Hoechst negative and positive fractions after FACS, were 12.67+/-2.24% and 1.17+/-6.13%, respectively (p<0.05). CONCLUSIONS: Putative corneal epithelial stem cells were efficiently isolated from limbal tissue using a trypsin/EDTA extraction and FACS. This technique may be very useful in tissue engineered stem cell therapy.
Trypsin/pharmacology ; Stem Cells/*cytology/drug effects ; Rabbits ; Limbus Corneae/*cytology/drug effects ; Humans ; Epithelium, Corneal/*cytology/drug effects ; Edetic Acid/pharmacology ; Cells, Cultured ; Cell Culture Techniques ; Cell Count ; Animals

PURPOSE: To analyze the isolating pattern of slow cycling cells as putative limbal epithelial stem cells (PLESCs) using Hoechst exclusive cell sorting. METHODS: Rabbits were injected with 5-bromo-2-deoxyuridine (Brd U) 1 month prior to be sacrificed. After obtaining limbal tissues, fluorescence-activated cells were sorted on a Coulter EPICS 753 after they had been incubated with Hoechst 33342 and propidium iodide. Two different methods were applied to sort PLESCs. Side-population(Sp) cells were obtained using gates with dichroic mirror to detect low Hoechst blue and red after verapamil was treated. Hoechst negative cells were obtained using gates exhibiting low Hoechst blue with a 424/44 BP filter. Brd U-retaining cells were counted and their sizes were evaluated in each gated sample to compare isolating pattern of PLESCs in each method. RESULTS: The percentages of Sp cells and of the Hoechst negative fraction were 0.96 +/- 0.79% and 16.01 +/- 13.60%, respectively(p=0.021). Homogeneity and density of the small cells were higher in Hoechst negative fraction than in Sp cells. The percentage of Brd U-retaining cells was 47.36 +/- 10.34% and 47.14 +/- 14.94% in Sp cells and Hoechst negative fraction, respectively(p>0.05), and they were 10 times higher than in non-Sp and Hoechst positive fraction(p=0.000). CONCLUSIONS: Hoechst negative exclusion without verapamil more efficiently isolated PLESCs than Sp did.
Cornea ; Epithelium ; Fluorescein* ; Propidium ; Rabbits ; Stem Cells* ; Verapamil