• Korean Journal of Clinical Laboratory Science
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• v.47 no.3
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• pp.132-139
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• 2015

Recently, $p16^{INK4a}$/Ki-67 dual immunostaining has been introduced as a new biomarker protocol for early detection of uterine cervical dysplasia and cancer in liquid-based cytology (LBC). We performed the $p16^{INK4a}$/Ki-67 dual immunostaining using a CINtec$^{(R)}$ PLUS kit in a total of 109 LBC cases of cervicovaginal smear and compared its results with those from LBC, HPV hybrid capture II (HC II) test and histological diagnosis. Expression of $p16^{INK4a}$ and Ki-67 was significantly associated with cases of LSIL or higher in cytological diagnosis and cases of cervical intraepithelial neoplasia (CIN) 1 or higher in histological diagnosis (p<0.001 and p<0.001, respectively). Among forty-six cases of atypical squamous cells of undetermined significance (ASCUS) in LBC, $p16^{INK4a}$ and Ki-67 was expressed in 31 (67.4%), which were positively associated with cases of CIN I lesion or higher in histology. The sensitivity of $p16^{INK4a}$/Ki-67 dual immunostaining for finding lesions of CIN 1 or higher was 89.0%, which was higher than LBC. The specificity was 73.5%, which was higher than that of the HC II test. Based on these results, the $p16^{INK4a}$/Ki-67 dual immunostaining method can be a useful diagnostic marker for improving the sensitivity of LBC and the specificity of HC II test.

• Korean Journal of Acupuncture
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• v.30 no.1
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• pp.56-63
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• 2013

Objectives : Effects of repetitive electroacupuncture(EA) on the pain behavior and activation of spinal glial cells were examined in the rat model of neuropathic pain. Methods : Twenty one adult male Sprague-Dawley rats were randomly assigned into 3 groups(control group, SP6 group, ST36+GB34 group). Neuropathic pain was induced by tight ligation of L5 spinal nerve. Mechanical and thermal hypersensitivity of hind paw were tested. Immunohistochemistry was performed in spinal cord L5/6 of all groups. EA was treated once in a day from the $5^{th}$ day after surgery. Results : EA treatments applied to ST36 and GB34 reduced significantly both of mechanical and thermal hypersensitivity after 3 times of treatment throughout the experiments. In the SP6 group, the analgesic effect was also shown after 7 times of treatment. Immunohistochemistry demonstrated inhibition of microglia and astrocyte activation in the spinal cord L5/6 dorsal horn in the ST36+GB34 group. Conclusions : The present results suggest that repetitive EA exert strong analgesic effect on neuropathic pain. These analgesic effects in neuropathic pain are associated with suppressing the activation of microglia and astrocyte.

• Tuberculosis and Respiratory Diseases
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• v.41 no.4
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• pp.339-353
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• 1994

Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

• Journal of Oral Medicine and Pain
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• v.36 no.4
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• pp.261-271
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• 2011

Osteoarthritis in patients with temporomandibular disorders(TMDs) induces pain, limitation of mouth opening, occlusal problems, and most commonly affects their life quality. Control method and progressive process of osteoarthritis are being extensively researched. The researchers focus on histologic changes, synovial changes, muscular and ligamental changes and observed reaction to pain. Therefore most of them developed the animal model for osteoarthritis in TMD patients. In this study, we applied several methods which induces osteoarthritis of temporomandibular joint(TMJ) in rats or mice. For locally induce osteoarthritis in TMJ, Monosodium iodoacetate(MIA) or interleukin-$1{\alpha}$(IL-$1{\alpha}$) were injected into TMJ joint space for 5 or 3 weeks. Other groups are chosen for osteoarthritis under systemic control including hormonal changes and aging. To observe cellular change, increased collagen, degenerative bony destruction and distribution of proteoglycans (PGs), safranin-O staining and Masson's trichrome staining were used.

• The Korean Journal of Pesticide Science
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• v.6 no.2
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• pp.96-104
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• 2002

This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.

• Neonatal Medicine
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• v.17 no.1
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• pp.44-52
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• 2010

Purpose : The aim of this study was to develop a model for necrotizing enterocolitis (NEC) in the neonatal rat using endotoxin and hypoxia, a plausible insult in a neonatal intensive care and to investigate the role of apoptosis as the underlying mechanism. Methods : Newborn rats were given oral endotoxin and intermittent 8% hypoxia$\pm$caspase inhibitor. The intestinal histology was evaluated using hematoxylin-eosin staining. Apoptosis was analyzed with TUNEL staining and by measuring the caspase 3 activity in the intestinal lysates. IEC-6 cells were assessed for apoptosis and the expression of Bax, Bcl-2, Fas and FasL was measured after treatment with endotoxin and hypoxia. Results : Oral endotoxin (5 mg/kg) and exposure to 8% hypoxia of 60-min duration twice induced human NEC-like lesions in the rat intestine. Intestinal tissue revealed increased apoptosis and caspase-3 activity. After caspase inhibitor treatment, the grades of both apoptosis and NEC were significantly reduced. IEC-6 cells exhibited increased apoptosis and caspase 3 activity after endotoxin and hypoxia treatment and significantly increased Bax/Bcl- 2 ratio compared to control cells. Conclusion : This neonatal rat model of NEC which was induced by oral endotoxin and intermittent hypoxia showed increased apoptosis of intestinal epithelial cells that was mediated by caspase 3 activation. Our model has a advantage in the study of NEC because the use of much more clinically plausible insults may provide a suitable model for the investigation of its pathophysiology and therapeutic trials.

• Journal of Veterinary Clinics
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• v.20 no.3
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• pp.278-285
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• 2003

This study was performed to compare early stage changes following the administrations of Conjugated Linoleic Acid (CLA), Carprofen and their combinations in the experimental canine cranial cruciate ligament rupture. Twenty five mongrel dogs were divided into five groups; Groups 1, 2, and 3 received a sectioning the cranial cruciate ligament (CCL) of the right stifle joint, and were administered CLA (250 mg/kg/day/orally), carprofen (4.4 mg/kg/day/orally), and their combinations for 4 weeks beginning 4 weeks postsurgery, respectively. Group 4 received sectioning CCL and no treatment. Group 5 was composed of unoperated normal dogs. The macroscopic observation of cartilage erosions on both the condyles and plateaus were evaluated, and the severity of the cartilage lesions and synovial inflammation was examined histologically at eight weeks after surgery. Histological examinations including hematoxylin and eosin stain, standard toluidine blue method, PAS technique and Masson trichrome technique, hematologic and radiographic evaluation were perfonned after experimental surgery. Slight yellowish discoloration of the surface was found in some of the experimental dogs. However macroscopic findings showed no significant differences among the groups. In radiographic findings, cranial displacement of the proximal tibia relative to the femoral condyles in all groups was observed but no significant differences among the groups was seen. This study showed that oral administration of CLA, carprofen and their combinations revealed no early stage change in the canine stifle joint following experimental rupture of the cranial cruciate ligament.

• Journal of Life Science
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• v.18 no.4
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• pp.580-584
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• 2008

$O^6-methylguanine-DNA$ methyltransferase (MGMT) is a DNA repair protein that protects cells against the carcinogenic effects of alkylating agents. The loss of MGMT expression was commonly known due to hypermethylation of CpG islands in its promoter region. We evaluated the expression of MGMT by immunohistochemistry in order to examine the relationship between loss of MGMT expression and clinicopathological characteristics in 74 Korean patients with non-small cell lung cancers. Loss of MGMT was detected in 25 (33.8%) of 74 cases. The loss of MGMT expression was frequently seen in the adenocarcinoma than in the squamous cell carcinoma (p=0.021). However, there was no significant differences between loss of MGMT expression and other clinicopathological characteristics, including age, gender, smoking status, tumor size, tumor T stage, and lymph node metastasis (p>0.05). In conclusion, loss of MGMT expression was related with the histologic type of lung cancer. Further methylation study of MGMT promoter is needed to evaluate the relationships with immunohistochemical expression of MGMT and to clarify the role of MGMT in lung cancer.

• Journal of the Korean Society of Radiology
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• v.8 no.7
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• pp.389-395
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• 2014

The research was carried out to develop a animal model of malignant brain tumor for the researches in glioblastoma multiform (GBM) diagnosis and therapy. C6 cells were transplanted into the right striatum of SD rat using stereotactic instrument for the development. The developed animal model was verified by MRI and H&E stain assay of anatomicohistological examination. The MRI observations showed that the tumor developed at the injection site at the 7 days after glioblastoma inoculation. At 14 days post inoculation, the tumor grew to a large volume occupying almost a half of the right cerebral hemisphere. It was confirmed that the expression of excessive mitosis and pleomorphism in anatomicohistological examination. The developed animal model must be necessary and useful tool for the in vivo level research in the development of the new modality for the diagnosis and therapy of brain cancer.

• Journal of Korean Society of Forest Science
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• v.37 no.1
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• pp.61-63
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• 1978

Both bark and wood sections obtained, by cutting with a razor blade, from living tissue of Cronartium ribicola cankers of Korean white pine (Pinus koraiensis) were transferred to a mixture of 2 parts of chloroform and 1 part of methanol, and the sections were stained using a modified lactophenol cotton blue. The formula for this staining is as follows: Lactic acid 20gm, phenol crystal 20gm, cotton blue 0.05gm, and 60% EtOH 100ml. The rust hyphae and haustoria were stained blue, and the wide hyphae with straight or curved haustoria could be distingwished from the pine tissue.