• Restorative Dentistry and Endodontics
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• v.16 no.1
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• pp.41-59
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• 1991

This study was performed to elucidate the distribution and correlation of immunoglobulin G subclasses with the degree of inflammation in the experimentally induced rat pulp and periapical pathoses. The pulp exposures were made in 108 mandibular 1st molars of 54 rats and the teeth were left open to the oral environment The animals were sacrified at 3, 7, 15, 30, 60 and 90 days after pulp exposure, and examined microscopically and radiographically Seventy one specimens were routinely sectioned at the thickness of 4 - $6{\mu}$ and stained with Hematoxylin - eosin for histologic examination, with toluidine blue for mast cells, and with the primary antibodies against rat IgG subclasses by using the Avidin - Biotin complex method. The following results were obtained: 1. As the degree of inflammation of rat pulp and periapeces intensified, the number of IgG subclass containing cells per unit area, especially IgG2a and IgG2c, decresased. 2. The IgG2c cells were most predominantly found in the lesions with slight inflammation, IgG1 cells in mild or severe inflammation, and IgG2a cells in moderate inflammation. 3. IgG subclass containg cells were more predominantly observed in the periapical granuloma than periapical abscess or cyst(p<0.01). 4. IgG2a containing cells were predominant in pulp inflammation, IgG1 containing cells in periapical granuloma, IgG2a cells and IgG1 cells in periapical abscess, and IgG2a cells were significantly predominant in periapical cyst. 5. The number of IgG subclass containing cells and mast cells in periapical tissue decreased with time lapse after pulp exposure. And correlation index between mast cells and IgG1, IgG2a, IgG2b was stastically high.

• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.605-621
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• 1993

The gingival hyperplasia refers to an increase in the size of the gingival tissue produced by an increase in the number of its component cells. In order to investigate the cellular change in epithelium and subepithelial tissue of noninflammatory gingival hyperplasia, the gingival tissues were surgically obtained from the patients with dilantin gingival hyperplasia and idiopathic gingival hyperplasia. The excised tissue samples were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.S.A.) and immunocytochemically processed by Avidin-Biotin peroxidase complex method for detecting proliferating cell nuclear antigen, tenascin and collagen type IV. Monoclonal mouse anti-human PCNA antibody(Oncogene Science, Uniondale, NY, U.S.A., 1 : 250,000), monoclonal mouse anti-human tenascin antibody(Chemicon-International Inc., Temecula, CA, U.S.A., 1:5,000), and monoclonal mouse anti-human collagen type IV(Dakopatts, Glostrup, Denmark, 1: 50) were used as primary antibodies. The results were as follows: 1. In non-inflammatory gingival hyperplasia, the positive reaction to proliferating cell nuclear antigen was localized in the basal cell layer of gingival epithelium and well-developed rete pegs. 2. The positive reaction to tenascin was shown in the connective tissue subjacent to basament membrane of gingival tissue, and especially strong positive reaction was noted in the tip portion of connective tissue projections. 3. The positive reaction to collagen type IV was localized along the basement membranes of gingival epithelium and blood vessels. The results suggest that connective tissue enlargement may affect the proliferation of gingival epithelium.

• Korean Journal of Veterinary Research
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• v.42 no.1
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• pp.7-20
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• 2002

This study deals with the distribution and morphological study of the neurons with the neuropeptide Y (NPY) in developing chick brain. The developing brains of Korean native chicks at embryonic days 8 (E8), E10, E12, E14, E16, E18, and E20 were used. The chicks were perfused with 4% paraformaldehyde through the left ventricle and aorta. The brains were removed and transferred into 30% sucrose, and then cut in a cryostat into $60{\mu}m$ in thickness. The sections were immunostained with free-floating and avidin-biotin complex (ABC) methods. The numerous neurons with NPY were first observed in nucleus rotundus of diencephalon at E8. In particular, neurons in nucleus rotundus had the well-developed processes. At E12, the neurons with NPY were distributed widely in diencephalon; nucleus septalis lateralis, medialis, nucleus magnocelluaris preopticus dorsalis, medialis, ventralis, nucleus preopticus medialis, nucleus dorsolateralis anterior thalami pars magnocellularis, and nucleus paraventricularis magnocellularis (paraventricular nucleus) except nucleus rotundus. From E12 to E20, positive neurons were oval-shaped, changed gradually into the spherical- and multipolar-shaped. The shapes of processes were also omnidirectional and the number of those were less than in telencephalon. As the chicks developed, the morphology of neurons with NPY showed the tendency to increase in their sizes and numbers.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.859-871
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• 1996

This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.501-509
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• 2021

More than half the world's population is thought to be infected with Helicobacter pylori. Although the majority of infected people are asymptomatic, H. pylori infection may cause gastric ulcers and deadly gastric cancer. Owing to the difficulty and invasiveness of current routine culture and diagnostic methods, a highly sensitive and specific noninvasive assay for H. pylori is of interest. This study highlighted the design and performance of a colorimetric magneto loop-mediated isothermal amplification (CM-LAMP) assay to detect H. pylori in spiked saliva samples. LF primers were coated on magnetic nanoparticles by carbodiimide-induced immobilization and functionally used for solid-phase amplification. During the LAMP reaction at 66℃, biotin-tagged FIPs were incorporated into LAMP amplicons. The colorimetric signal developed after the addition of NeutrAvidin horseradish peroxidase conjugate (NA-HRP) and ABTS. None of the tested microorganisms, including closely related bacteria, was shown positive by the CM-LAMP assay except H. pylori isolates. This novel platform was highly specific and 100-fold more sensitive (40 CFU/ml or 0.2 CFU per reaction) than the PCR and conventional LAMP assays for the detection of H. pylori in spiked saliva. Our results demonstrated the feasibility of using this noninvasive molecular diagnostic test to detect H. pylori in saliva samples.

• Radiation Oncology Journal
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• v.19 no.2
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• pp.181-189
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• 2001

Purpose : Changes in the balance between MMP and TIMP can have a profound effect on the composition in the extracellular matrix (ECM) and affect various cellular functions including adhesion, migration, differentiation of cells, and fibrosis and invasion and metastasis of cancer cells. Radiation therapy is a popular treatment modality for benign and malignant tumor, but the study for radiation effect on MMP and TIMP is scarce. In the current study, we have examined the expression of TIMP in fibrosis-prone (C57BL/6) mice after radiation. Methods and Materials : Adult female mice of $10\~12$ weeks were used. The whole body were irradiated using a Varian CL-4/100 with 2 and 10 Gy. Immunohistochemical staining was peformed according to Avidin Biotin complex method and evaluated by observing high power field. For TIMP-1, TIMP-2 antibodies, reactivity was assessed in the parenchymal cell and in the stromal cell. The scale of staining was assessed by combining the quantitative and qualiative intensity of staining. Results : TIMP-1 immunoreactivity did not change in lung. But, in liver, TIMP-1 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell. in kidney, TIMP-1 immunoreactivity was localized in cytoplasm of some tubular cell. Temporal variations were not seen. Dose-response relationship was not seen except kidney. TIMP-2 immunoreactivity in lung was a score (++) at 0 Gy and elevated to a score (+++) at 2 Gy. TIMP-2 immunoreactivity was a score (++) in liver at 0 Gy. TIMP-2 immunoreactivity was localized in cytoplasm of hepatocyte and Kupffer cell as same as patterns of TIMP-1 immunoreactivity. The TIMP-2 immunoreactivity in liver was elevated to (+++) at 2 Gy. Immunoreactivity to TIMP-2 in kidney was a score (+++) at 0 Gy and was not changed at 10 Gy. The score of TIMP-2 immunoreactivity was reduced to (++) at 2 Gy. TIMP-2 immunoreactivity was confined to tubules in kidney. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIMP-2 immunoreactivity was not seen. Conclusions : Differences between intensity of expression of TIMP-1 and TIMP-2 in each organ was present. Expression of TIMP was localized to specific cell in each organ. Irradiation increased TIMP-1 immunoreactivity in the liver and the kidney. Irradiation increased TIMP-2 immunoreactivity in the lung. But, in the liver and the kidney, TIMP-2 expression to radiation was irregular. Temporal variation of TIMP-2 immunoreactivity was irregular. Dose-response relationship of TIHP-2 immunoreactivity was not seen. In the future, we expect that the study of immunohistochemical staining of longer period of postirradiation and quantitative analysis using western blotting and northern blotting could define the role of TIMP in the radiation induced tissue fibrosis.

• Journal of Chest Surgery
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• v.39 no.11 s.268
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• pp.828-837
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• 2006

Background: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1(HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing $O_2$ delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-$1{\alpha}$ has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-$1{\alpha}$ on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-$1{\alpha}$ expression on angiogenetic factors, relationship between the tumor proliferation and HIF-$1{\alpha}$ expression, interaction of HIF-$1{\alpha}$ expression and p53, and relationship between HIF-$1{\alpha}$ expression and clinico-pathological prognostic parameters were investigated. Material and Method: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-$1{\alpha}$, VEGF(vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex(ABC) immunohistochemistry. Relationship between the HIF-$1{\alpha}$ expression and VEGF, p53 overexpression and correlation between the HIF-$1{\alpha}$ expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. Result: HIF-$1{\alpha}$ expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma(40.7%). High HIF-$1{\alpha}$ expression was significantly associated with several pathological parameters, such as pathological TMN stage(p=0.004), pT stage(p=0.020), pN stage (p=0.029), and lymphovascular invasion(p=0.019). High HIF-$1{\alpha}$ expression was also significantly associated with VEGF immunoreactivity(p<0.001), and aberrant p53 expression(p=0.040). but was marginally associated with Ki-67 labeling index(p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-$1{\alpha}$ expression was worse than that of patients with low-expression(p=0.002). High HIF-$1{\alpha}$ expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. Conclusion: It is suggested that high HIF-$1{\alpha}$ expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-$1{\alpha}$ expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung carcinoma.

• Journal of Korean Neurosurgical Society
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• v.30 no.4
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• pp.443-450
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• 2001

Objective : The cyclin-dependent kinase inhibitor $p27^{kip1}$ protein is a negative regulator of the cell cycle, and its degradation is required for entry into the S phase. Loss of $p27^{kip1}$ expression has been reported to be associated with aggressive behavior in a variety of tumors of epithelial and lymphoid origin. However, its association with various astrocytic tumors has not been clearly demonstrated. We studied to investigate the relationship of $p27^{kip1}$ expression with the biological behavior of astrocytic tumors in addition to study on the role of $p27^{kip1}$ in the tumorigenesis of these tumors. Patients and Methods : From 1990 to 1998, a total of 29 astrocytic tumor of all grades obtained by operative resection were included for evaluation. We studied the expression of $p27^{kip1}$ protein immunohistochemical assay in astrocytic tumors and compared the findings with the clinicopathologic parameters. Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by the avidin-biotin-peroxidase complex method. According to WHO classification, all cases were divided into astrocytomas(4 cases), anaplastic astrocytomas(9 cases), and glioblastomas(16 cases) by 3 pathologists. Clinical information was obtained from medical records, and others such as location and size of tumors from imaging studies. Results : Mean $p27^{kip1}$ protein labeling indexes(LI, mean${\pm}$standard deviation) of astrocytomas, anaplastic astrocytomas, and glioblastomas were $80.6{\pm}9.1$, $63.6{\pm}21.0$, and $28.9{\pm}18.7$, respectively, and were inversely correlated with grade of glial tumors(p<0.0001). Mean $p27^{kip1}$ protein LI in the recurrent group was lower than that in the nonrecurrent group, but there was no significant difference statistically(p=0.464). Additionally, $p27^{kip1}$ protein expression did not show any significant relationship to other prognostic factors such as age(p=0.1643), tumor size(p=0.8), or location(p=0.8). Conclusion : These results suggested that reduced expression of $p27^{kip1}$ protein may play a important role in the malignant transformation process of astrocytic tumor cells.

• Journal of Korean Neurosurgical Society
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• v.30 no.4
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• pp.430-436
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• 2001

Objective : The proliferative potential of intracranial glioma affects the histological malignancy and prognosis of patients with these tumors. In this study, we present the relationship between MIB-1 labeling index(LI) and clinical variables which might play the major role in determining the prognosis of patient with astrocytic tumors. Patients and Methods : Excised tumor specimens from a total of 52 patients were stained to detect monoclonal MIB-1-Ki-67 antibody by avidin-biotin complex immunohistochemistry. The MIB-1 LI was evaluated with histological grades, demograpghic data, and survival time. The statistical significance of their correlation was analyzed by Pearson correlation test. Results : The 52 patients included 30 male patients and 22 female patients. The tumors according to the criteria of the World Health Organization(WHO) classification were verified as pleomorphic xanthoastrocytoma in one, pilocytic astrocytomas 4, astrocytomas 1, anaplastic astrocytomas 3, and glioblastomas 31. MIB-1 LI in astrocytic ttumors showed no correlation with age and gender. However, the patients under 10 years had the longest survival time, whereas short survival time was observed in the older patients. The mean MIB-1 LI of different tumor grades were as follows : pleomorphic xanthoastrocytoma, $4.40{\pm}0.00$ ; pilocytic astrocytoma, $4.53{\pm}3.09$ ; astrocytoma, $5.50{\pm}6.03$ ; anaplastic astrocytoma, $12.68{\pm}12.50$ ; Glioblastoma, $21.31{\pm}19.63$. Although the levels of MIB-1 LI were varied in individual tumors, the MIB-1 LI was increased in parallel with the histological grades. Glioblstomas showed significantly higher MIB-1 LI compared with that of anaplastic astrocytomas and low grade astrocytomas (p = 0.001). The mean survival time of entire group of patients was also well correlated with MIB-1 LI in astrocytic tumors(p = 0.015). Moreover, the mean survival time of the entire group of patients with Lis < 10 was $125.33{\pm}113.57weeks$, and the mean survival of those with $Lis{\geq}10$ was $60.71{\pm}62.58weeks$. This difference was also statistically significant(p = 0.004). Conclusion : The results of this study suggest that MIB-1 LI correlates with histological grades and might play a significant role in predicting the survival of patients with astrocytic tumors.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.121-130
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• 1994

Expression of gonadotropin releasing hormone(GnRH) has been described in the rat ovary. It remains, however, unkown whether GnRH is synthesized as a prohormone. Therefore, this study was performed to verify the expression of pro-GnRH by in situ hybridization and further to investigate the effect of gonadotropin on GnRH or GnRH mRNA in rat ovary by immunohistochemical and in situ hybridization techniques. Adult female Sprague-Dawely rats were used and the estrous cycle was synchronized by intraperitoneal injection of pregnant mare's serum gonadotropin(PMSG). Ovaries were fixed with 4% paraformaldehyde and embedded with G.C.T. compound and cut by cryostat. For immunohistochemistry, avidin-biotin peroxidase complex(ABS) method was employed and for in situ hybridization, $^{35}S$-end labeled oligonucleotide was used and followed by autoradiography. By in situ hybridization using GnRH oligomer and GAP(GnRH associated protein) oligomer, GnRH mRNA and GAP mRNA were co-localized in the fullicular cells, luteal cells, interstitial cells and theca cells. GnRH or GnRH mRNA signals in the ovary increased by human chorionic gonadotropin(hCG) injection. At the 3 and 6 hrs after hCG injection, the number of GnRH and GnRH mRNA containing cells increased rapidly and the density of GnRH and GnRH mRHA culminated at 9 hrs after heG injection. With the follicular development, the high expression of GnRH and GnRH mRNA was also observed within the follicles. After ovulation, the density of GnRH or GnRH mRNA decreased in the follicles but increased in the corpus lutea.