To achieve optimal decalcification in tissue and tissue preservation, we have tried magnetic field method and made some promising results. We used pulsed unipolar magnetic field obtained by rectification of 250 V-60 cycle, A.C. As a new method of bony decalcification, using 5% nitric acid, 10% formic acid and 10% formic acid+3% hydrochloric acid solutions, experimental groups were decalcified in the center of the magnetic field. The concentration of calcium ion in the decalcifying solution was measured by calcium-oxalate turbidity test by photometry method, and direct visualization of calcium radiopacity was obtained by soft X-ray view during the decalcification process. The pH change during decalcification was continuously checked and needle penetration method was also used. All the decalcification solution used in this study showed accelerated effect of bony decalcification in the strong magnetic field. Among them 5% nitric acid produced complete decalcification for the medium size bony specimen (less than 10x10x10 mm) within 24 hours, and the histologic feature was almost free of acid-chemical degeneration. The pH of all the decalcification solutions decreased in the strong magnetic field, maximum within 4~6 hours, and kept strong acidity throughout the decalcification procedure. After removal of the magnetic field the pH of all the decalcification solution returned to their original values after 24 hours. It was presumed that the cause of the accelerated decalcification in the magnetic field was due to combined effects of the rapid increase of acidity and the increased molecular resonance to stimulate the ionization of mineral elements.

Ceruminous glands are specialized apocrine glands located in the external auditory canal (EAC). Pleomorphic adenoma (PA) of the EAC is derived from these ceruminous glands. Tumors arising from these ceruminous glands are rare. Furthermore, ceruminous PA of the EAC is extremely rare. About 35 cases have been reported to date in the English literature, and only 4 cases have been reported in Korea. There are several controversial issues about these rare tumors such as nomenclature, histogenesis and classification. We report here on two cases of ceruminous PA and review the cases in the Korean literature.
Adenoma ; Adenoma, Pleomorphic* ; Apocrine Glands ; Classification ; Ear Canal* ; Korea

Although segmental or subsegmental atelectasis may occur during anesthesia, mucous plugging of a mainstem bronchus has been uncommonly reported in anesthetized patients with chronic respiratory disease. However, pulmonary atelectasis following mucous plugging may rarely result normal patients. We report this case of an allegedly healthy patient was developed a left main stem bronchus obstruction, resulting in subsegmental collapse of left lower lung after the induction of general anesthesia.
Anesthesia* ; Anesthesia, General ; Bronchi ; Humans ; Lung ; Pulmonary Atelectasis

We experienced a case of idiopathic crescentic glomerulonephritis in a 10-year-old girl who was admitted to our hospital due to gross hematuria and oliguria for 2 months. The diagnosis was based on the rapidly progressive clinical course to chronic renal failure, positive p-ANCA test and light microscopic findings of diffuse crescents formation in about 80% glomeruli of renal tissue obtained by percutaneous renal biopsy. Our case was treated with high dose methylprednisolone ( 30mg/Kg) pulse therapy whenever its clinical course was aggravated for several times. After 2 years 6months later, she has been treated on peritoneal dialysis. A review of literatures was also presented briefly.
Antibodies, Antineutrophil Cytoplasmic ; Biopsy ; Child ; Diagnosis ; Female ; Glomerulonephritis* ; Hematuria ; Humans ; Kidney Failure, Chronic ; Methylprednisolone ; Oliguria ; Peritoneal Dialysis

OBJECTIVE: This study was to establish a reproducible differentiation system from the parthenogenetic mouse embryonic stem (P-mES02) cells into functional cardiomyocytes like as in vitro fertilization mouse embryonic stem (mES01) cells. MATERIALS AND METHODS: To induce differentiation, P-mES02 cells were dissociated and aggregated in suspension culture environment for embryoid (EB) formation. For differentiation into cardiomyocytes, day 4 EBs were treated with 0.75% dimethyl sulfoxide (DMSO) for another 4 days (4-/4+) and then were plated onto gelatin-coated dish. Cultured cells were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. This frequency was compared with the results of DMSO not treated P-mES02 group (4-/4-) and mES01 groups (4-/4+ or 4-/4-). For confirm the generation of cardiomyocytes, beating cell masses were treated with trypsin-EDTA, dispersed cells were plated onto glass coverslips and incubated for 48 h. Attached cells were fixed using 4% paraformaldehyde and incubated with specific antibodies (Abs) to defect cardiomyocytes (anti-sarcomeric alpha-actinin Ab, 1: 100; anti-cardiac troponin I Ab, 1: 2000) for 1 h. And the cells were finally treated with FITC or TRITC labelled 2nd Abs, respectively, then they were examined under fluorescence microscopy. RESULTS: Rhythmically contracting areas in mES01 or P-mES02 cells were firstly appeared at 9 or 10 days after EBs plating, respectively. The highest cumulative frequency of beating EBs was not different in both treatment groups (mES01 and P-mES02, 4-/4+) with the results of 61.3% at 13 days and 69.8% at 15 days, respectively. Also the contracting duration of individual beating EBs was different from minimal 7 days to maximal 53 days. However, DMSO not treated groups (mES01 and mES02,4-/4-) also had contracting characteristics although their frequency was a few compared to those of DMSO treated groups (6.0% and 4.0%). Cells recovered from the spontaneously contracting areas within EBs in both treated groups were stained positively with muscle specific anti-sarcomeric alpha-actinin Ab and cardiac specific anti-cardiac troponin I Ab. CONCLUSION: This study demonstrated that the P-mES02 cell-derived cardiomyocytes displayed similarly structural properties to mES01 cell-derived cardiomyocytes and that the DMSO treatment enhanced the cardiomyocytes differentiation in vitro.
Actinin ; Animals ; Antibodies ; Cells, Cultured ; Dimethyl Sulfoxide ; Embryonic Stem Cells* ; Fertilization in Vitro ; Fluorescein-5-isothiocyanate ; Glass ; Mice* ; Microscopy, Fluorescence ; Myocytes, Cardiac* ; Troponin I

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

88 workers exposed to cadmium were examined at the 12 factories using or producing cadmium in order to know the present state of cadmium exposure and renal dysfunction in 1992. Cadmium in blood and in urine were measured and compared by the 3 exposure level of cadmium in air. Cadmium in blood of low, moderate and high exposure group were 2.5, 3,8 and 7.6 microgram/L, respectively. Cadmium in urine were 1.8(1.3), 3.8(2,6) and 7.9 microgram/L(6.1 microgram/g creatinine) , resrectively. However, there was no relationship between Urinary cadmium and beta(2)-microglobulin. Cumulative exposure estimate (CEE) was calculated by multiplying the mean ambient cadmium level of the factory and working duration. CEE has a high correlation with cadmium in blood and urine, but no relation to beta(2)-microglobulin. Because working durations were relatively shorter than European workers', the highest CEE was just 300 microgram. year/m(3), which was not enough to induce renal tubular dysfunction. This study, however, suggested the possibility that renal tubular dysfunction caused by cadmium could be happened in Korea in the near future.
Cadmium* ; Korea ; Occupations*

No abstract available.
Animals ; Constitution and Bylaws* ; Mice* ; Oocytes*

OBJECTIVE: This study was to establish the human embryonic stem (ES) cells derived from frozenthawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. METHODS: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. RESULTS: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and beta-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. CONCLUSION: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.
Humans

PURPOSE: We conducted a prospective non-randomized clinical study to evaluate the efficacy and toxicity of the preoperative concurrent chemoradiotherapy for locally advanced unresectable rectal cancer. MATERIALS AND METHODS: Between January 1995 and June 1998, 37 consecutive patients with locally unresectable advanced rectal cancer were entered into the study. With 3- or 4- fields techniuqe, a total of 45 Gy radiation was delivered on whole pelvis, followed by 5.4 Gy boost to the primary tumor in some cases. Chemotherapy was done at the first and fifth week of radiation with bolus i.v. 5-Fluorouracil (FU) 370~450 mg/m2, days 1~5, plus Leucovorin 20 mg/m2, days 1~5. Of 37 patients, 6 patients did not receive all planned treatment course (refusal in 4, disease progression in 1, metastasis to lung in 1). Surgical resection was undergone 4~6 weeks after preoperative concurrent chemoradiotherapy. RESULTS: Complete resection rate with negative margins was 94% (29/31). Complete response was seen in 7 patients (23%) clinically and 2 patients (6%) pathologically. Down staging of tumor occured in 21 patients (68%). Treatment related toxicity was minimal except grade III & IV leukopenia in 2 patients, respectively. CONCLUSION: Preoperative concurrent chemoradiotherapy in locally advanced rectal cancer was effective in inducing down staging and complete resection rate. Treatment related toxicity was minimal. Further follow up is on-going to determine long term survival following this treatment.
Chemoradiotherapy* ; Disease Progression ; Drug Therapy ; Fluorouracil ; Follow-Up Studies ; Humans ; Leucovorin ; Leukopenia ; Lung ; Neoplasm Metastasis ; Pelvis ; Prospective Studies ; Radiation Dosage ; Rectal Neoplasms*