1.Analysis of the Gene Expression by Laser Captured Microdissection (I): Minimum Conditions Required for the RNA Extraction from Oocytes and Amplification for RT-PCR.
Chang Eun PARK ; Jung Jae KO ; Kwang Yul CHA ; Kyung Ah LEE
Korean Journal of Fertility and Sterility 2001;28(3):183-190
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
2.A Case of Leiomyoblastoma.
Ok Jae LEE ; Jee Hoon LEE ; Woo Song HA ; Kyung Hyuck KO
Korean Journal of Gastrointestinal Endoscopy 1996;16(6):989-993
Leiomyoblastoma is uncommon and has been known as a neoplasm of smooth muscle origin. However, with recent pragress in immunohistochemical staining techniques, many clinicopathological discrepancies have been pointed out about the origin of leiomyoblastoma. We present here a case of gastric leiomyoblastoma did not expressed desmin and neuron specific enolase, and was thought to be of unknown derivation.
Desmin
;
Leiomyoma, Epithelioid*
;
Muscle, Smooth
;
Phosphopyruvate Hydratase
3.A Case of Leiomyoblastoma.
Ok Jae LEE ; Jee Hoon LEE ; Woo Song HA ; Kyung Hyuck KO
Korean Journal of Gastrointestinal Endoscopy 1996;16(6):989-993
Leiomyoblastoma is uncommon and has been known as a neoplasm of smooth muscle origin. However, with recent pragress in immunohistochemical staining techniques, many clinicopathological discrepancies have been pointed out about the origin of leiomyoblastoma. We present here a case of gastric leiomyoblastoma did not expressed desmin and neuron specific enolase, and was thought to be of unknown derivation.
Desmin
;
Leiomyoma, Epithelioid*
;
Muscle, Smooth
;
Phosphopyruvate Hydratase
4.Clinical Studies on Ventricular Septal Defect with Septal Aneurysm.
Jae Kon KO ; Jung Yun CHOI ; Yong Soo YUN ; Chang Yee HONG ; Kyung Mo YEON
Journal of the Korean Pediatric Society 1986;29(5):40-45
No abstract available.
Aneurysm*
;
Heart Septal Defects, Ventricular*
5.Immunohistochemical and Ultrastructural Studies of Gastric Smooth Muscle Tumor.
Hyang Mi KO ; Kyung Soo KIM ; Jae Hyuk LEE ; Woo Sik JUHNG ; Sang Woo JUHNG
Korean Journal of Pathology 1996;30(3):245-254
To evaluate the differentiation status of smooth muscle in gastric stromal tumors which were negative for S-100 protein, immunohistochemistry using desmin, actin, myosin and vimentin was performed in 14 cases of gastric smooth muscle tumors. Ultrastructural Examination was also performed. For comparison a case of leiomyoma of the esophagus, a case of the sigmoid colon, 10 cases of the uterus were also examined. The results obtained were as follows. All gastric smooth muscle tumors showed vimentin-positivity. Six of 14 gastric smooth muscle tumors, (5 of 8 leiomyoma and 1 of 4 leiomyosarcoma) showed positivity for desmin, actin, and myosin(42.9%). All esophageal, colonic, and uterine leiomyomas showed diffuse positive reaction for desmin, actin, and myosin. Vimentin positivity was also noted in leiomyoma of the colon and uterus. Ultrastructurally, a few cells in the gastric stromal tumors had scattered microfilaments with dense bodies, subplasmalemmal dense plaques, and micropinocytic vesicles. However, most of the tumor cells did not have any of the ultrastructural features of smooth muscle differentiation. Leiomyomas of the esophagus and uterus showed many cytoplasmic microfilaments with dense bodies. These results suggest that most of the benign and malignant tumor cells of gastric stromal tumors have features of the undifferentiated cells, immunohistochemically as well as ultrastructurally, although a few cells have. It is speculated that most gastric stromal tumors may have lost their smooth muscle differentiation.
6.Endoscopic Therapy for Early Rectal Cancer: Piecemeal polypectomy and strip biopsy resection technique.
Ok Jae LEE ; Mee Jeong SOHN ; Young Chai KIM ; Jung Hee LEE ; Kyung Hyuk KO
Korean Journal of Gastrointestinal Endoscopy 1995;15(1):119-125
The incidence of colorectal cancer has been increased gradually and it was about 6.9% of all malignancies in Korea. Early diagnosis was recognized as the most important factor influencing the prognosis of colorectal cancer and the incidence of early colorectal cancer was increasing. Thus great change was observed in the treatment of early colorectal cancer, endoscopic therapy as well as curative surgical resection is being accepted. We report a case of effective and safe endoscopic therapy for early rectal cancer in 5S-year-old male patient. He presented with rectal prolapse and bleeding of a year's duration. A large polypoid mass with fine nodular surface and thick, short stalk was seen in the rectum at 4cm above the anal verge during flexible sigmoidoscopy. After the histologic examination of specimen obtained by bite biopsy, CT scan of pelvic cavity and ultrasonogram of abdomen, the tumor was removed safe and completely by 2 sessions of piecemeal polypectomies and strip biopsy. He has been well without symptoms and signs of recurrence of rectal cancer for 20 months.
Abdomen
;
Biopsy*
;
Colorectal Neoplasms
;
Early Diagnosis
;
Hemorrhage
;
Humans
;
Incidence
;
Korea
;
Male
;
Prognosis
;
Rectal Neoplasms*
;
Rectal Prolapse
;
Rectum
;
Recurrence
;
Sigmoidoscopy
;
Tomography, X-Ray Computed
;
Ultrasonography
7.A case of systemic progressive sclerosis after cosmetic surgery with silicone gel.
Won KIM ; Kyung Suk LEE ; Chang Hwan LEE ; Jae Ki KO
Korean Circulation Journal 1993;23(3):474-480
Human Adjuvant Disease(HAD) is a connective tissue disease occuring after cosmetic surgery with a foreign substance. It was first reported by Miyoshi in 1964. There have been specific connective disorders such as systemic lupus erythematosus, rheumatoid arthritis, progressive systemic sclerosis and mixed connective tissue disease. Scleroderma is the single most commonly reported disease. A 55-year-old woman was seen for evaluation of dypnea. Four years previously, she underwent cosmetic surgery with silicone injected directly into eyelid, forehead, both hand and monspubis. The patient presented the classic feature of systemic sclerosis and pericardial effusion. Following pericardiocentesis, she felt somewhat better and became less dyspneic. Seventeen days later, however, she progressively deteriorated and died in spite of intensive medical therapy and supportive care.
Arthritis, Rheumatoid
;
Connective Tissue Diseases
;
Eyelids
;
Female
;
Forehead
;
Hand
;
Humans
;
Lupus Erythematosus, Systemic
;
Middle Aged
;
Mixed Connective Tissue Disease
;
Pericardial Effusion
;
Pericardiocentesis
;
Scleroderma, Diffuse
;
Scleroderma, Systemic
;
Sclerosis*
;
Silicone Gels*
;
Surgery, Plastic*
8.A Case of Infantile Polycystic Kidney.
Il Young KO ; Chang Ho JUNG ; Jin Berm SONG ; Kyung Young SEO ; Jae Sik SHIM
Korean Journal of Obstetrics and Gynecology 1999;42(9):2115-2117
The infantile polycystic kidney disease is rare fetal urinary tract anomaly. It is inherited with an autosomal recessive pattern and recurrence rate is 25%. The gene locus is on chromosome 6p. The pathogenesis of infantile polycystic kidney is the primary defect of the collecting ducts. The ultrasonographic finding of infantile polycystic kidney is oligohydramnios, bilaterally symmetrical enlarged kidneys with maintenance of their reinform shape. The differential diagnosis with adult polycystic kidney disease is important. The massive enlargement of the kidneys is rarely seen in adult polycystic kidney disease and the examination of the parents and other members of the family is helpful to confirm the adult polycystic kidney disease. If there is severe renal involvements, stillbirth or neonatal death secondary to pulmonary hypoplasia would be developed. If it were diagnosed before viability, termination of pregnancy is recommended. In a fetus at risk, diagnosed after viability, pregnancy termination is also recommended since this condition is uniformly fatal. We present a case of infantile polycystic kidney.
Diagnosis, Differential
;
Female
;
Fetus
;
Humans
;
Kidney
;
Oligohydramnios
;
Parents
;
Polycystic Kidney Diseases*
;
Polycystic Kidney, Autosomal Dominant
;
Pregnancy
;
Recurrence
;
Stillbirth
;
Urinary Tract
9.Immunohistochemical Study of Gastrointestinal Stromal Tumors.
Jung Weon SHIM ; Hye Jae CHO ; Ill Hyang KO ; Ok Kyung KIM
Korean Journal of Pathology 1991;25(2):93-103
Historically, gastrointestinal stroma tumors (GIST) have been considered as smooth muscle tumors, but the controversy over this histogenesis is provoked due to various results with utilizing immunohistochemical methods. In andeffort to further clarify the histogenesis of GIST, we performed the immunohistochemical study, as well as histopathologic reexamination, of 24 cases, all diagnosed as smooth muscle tumors of gastrointestinal tract, from Seoul Paik Hospital and Ewha University Hospital between 1980 and 1989, and the main results were as follows; 1) In the histopathologic features by light microscopic study, 11 benign and 13 malignant lesions (including one high grade malignancy and 12 low-grade malignant lesions) were disclosed. 2) In the immunohistochemical study, all tumors showed Vimentin positivity (100%), but no tumor showed S-100 protein positivity (0%), and 7 cases (29.1%) showed Desmin positivity. Positive reaction for Desmin made it possible to suggest that the histogenesis of GIST be in smooth muscle, and neurogenic origin would be excluded by all negativity for S-100 protein. In summary, we would like to conclude that GIST would be smooth muscle tumors on account of their morphological characteristics and their intramural location, but most of them appear poorly differentiated by immunohistochemical method.
10.A Clinical Study on the Treatment of Femoral Shaft Fractures in Children: Immediate Hip Spica Cast Immobilization
Hyoun Oh CHO ; Chul Eun KO ; Kyung Duk KWAK ; Sung Do CHO ; Jae Hak LEE
The Journal of the Korean Orthopaedic Association 1988;23(3):707-712
The traditional treatment for the femoral shaft fractures in children is closed reduction by continuous traction, usually for 2–3 weeks, until the fracture has achieved partial stability, followed by spica cast immobilization. Fifty three cases, from 3 to 10 years of age, has been treated with closed reduction and immediate application of hip spica csst; all of them were closed fractures of femoral shaft and were not associated with head, abdominal, or vascular injuries. Any angulations in excess of acceptable ranges were corrected with wedging the cast; any overridings in excess of acceptable ranges were corrected by skeletal traction for 1 week, followed by incorporation of the pin to the cast. The resultant overriding and angulation of fragments were within acceptable ranges, and complications were not significant.This method of treatment not only is likely easy in nursing care and comfortsble to the patients but also has some advantages of short stay in the hospital, better utilization of hospital beds, and costsaving. This immediate hip spica cast immobilization seems to be a good method of treatment for the femoral shaft fracture in children.
Child
;
Clinical Study
;
Fractures, Closed
;
Head
;
Hip
;
Humans
;
Immobilization
;
Methods
;
Nursing Care
;
Traction
;
Vascular System Injuries