• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4545-4548
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• 2013

The objective of this study is to assess tissue expression of CA-125 and HE4 protein in primary benign and malignant epithelial tumours of the ovary and correlate with serum CA-125 levels. A total of 100 formalin-fixed, paraffin embedded sections of ovarian tumours which included serous adenoma (11), mucinous adenoma (42), serous carcinoma (20), mucinous carcinoma (12) and endometrioid carcinoma (15), histologically diagnosed between $1^{st}$ January 2004 to $31^{st}$ December 2012 at the University Malaya Medical Centre, were stained for HE4 (rabbit polyclonal antibody, Abcam, UK) and CA-125 (mouse monoclonal antibody clone: OC125, Cell Marque Corporation, Rocklin, California, USA). Pre-operative serum CA-125 levels were obtained from the laboratory information system. Immunoscore (I score) for HE4 and CA-125 was given based on the intensity of staining and percentage of positive tumour cells and considered significant when it was >50 (intensity of staining multiplied by percentage of positive tumour cells). Serum CA-125 levels were compared with the I score of HE4 and CA-125 in tissues. We noted that the CA-125 levels in serum and tissues were significantly raised in malignant compared to benign ovarian tumours (p value<0.05). Tissue expression of HE4 protein was also significantly raised in malignant tumours compared to benign tumours (p value<0.05). We conclude that HE4 can be a useful tissue immunomarker in addition to CA-125.

• Restorative Dentistry and Endodontics
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• 제21권2호
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• pp.677-687
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• 1996

This study was conducted to evaluate the tissue responses histologically to three root canal cements : Sealapex, AH-26, and zinc oxide-eugenol cement. Twelve white female Sprague-Dawley rats, weighing between 350 and 400 gm, were anesthetized with an intraperitoneal injection of Ketamine hydrochloride(0.4 ml). After shaving the sites selected(left and right scapular areas, left and right pelvic areas), the animal's backs were scrubed with soap and water, and sterilized with absolute alcohol. Each material was mixed to a thin consistency to flow out easily through a 24-guage needle, and loaded into a sterile, disposable plastic 1-ml syringe. All of the rats were injected subcutaneously with 0.1 ml of the three test sealers. Normal saline was used as a control. Animals were sacrificed after 48hr, 1, 4, and 12 weeks by overanesthetization using jars containing anesthetic ether. The tested sites were surgically removed with the surrounding tissue and fixed with 10% formalin. After 48 hours specimens were embedded in paraffin, sectioned to an average thickness of $6{\mu}m$ thick, stained with hematoxylin-eosin. The slides were examined under the light microscope. The results were obtained as follows 1. All material except the control showed various degree of inflammation on 48 hr. 2. Sealapex : In early stage, severe inflammatory cell infiltration was observed. At the 4th weeks observation, graunlomatous tissue with macrophage and foreing body giant cells containing many dark particles in their cytoplasm was observed. 3. AH-26 : Mild inflammatoy reaction was observed with AH-26 throughout the experimental period. 4. Zinc oxide-eugenol cement : Severe inflammatory cell infiltration, necrosis along the material, edema could be seen in early stage. Zinc oxide-eugenol cement maintained a moderate/severe reaction throughout the experimental period.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.245-252
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• 2014

Human METCAM/MUC18, a cell adhesion molecule (CAM) in the immunoglobulin-like gene super family, plays a dual role in the progression of several epithelium cancers; however, its role in the nasopharyngeal carcinoma (NPC) remains unclear. To initiate the study we determined human METCAM/MUC18 expression in tissue samples of normal nasopharynx (NP), NPCs, and metastatic lesions, and in two established NPC cell lines. Immunoblotting analysis was used for the determination in lysates of frozen tissues, and immunohistochemistry (IHC) for expression in formalin-fixed, paraffin-embedded tissue sections of 7 normal nasopharynx specimens, 94 NPC tissue specimens, and 3 metastatic lesions. Human METCAM/MUC18 was expressed in 100% of the normal NP, not expressed in 73% of NPC specimens (or expressed at very low levels in only about 27% of NPC specimens), and expressed again in all of the metastatic lesions. The level of human METCAM/MUC18 expression in NPC tissues was about one fifth of that in the normal NP and metastatic lesions. The low level of human METCAM/MUC18 expression in NPC specimens was confirmed by a weak signal of RT-PCR amplification of the mRNA. Low expression levels of human METCAM/MUC18 in NPC tissues were also reflected in the seven established NPC cell lines. These findings provided the first evidence that diminished expression of human METCAM/MUC18 is an indicator for the emergence of NPC, but increased expression then occurs with metastatic progression, suggesting that huMETCAM/MUC18, perhaps similar to TGF-${\beta}$, may be a tumor suppressor, but a metastasis promoter for NPC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1771-1774
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• 2014

The rhabdomyosarcoma (RMS) is the most common type of soft tissue tumor in children and adolescents; yet only a few screens for oncogenic mutations have been conducted for RMS. To identify novel mutations and potential therapeutic targets, we conducted a high-throughput Sequenom mass spectrometry-based analysis of 238 known mutations in 19 oncogenes in 17 primary formalin-fixed paraffin-embedded RMS tissue samples and two RMS cell lines. Mutations were detected in 31.6% (6 of 19) of the RMS specimens. Specifically, mutations in the NRAS gene were found in 27.3% (3 of 11) of embryonal RMS cases, while mutations in NRAS, HRAS, and PIK3CA genes were identified in 37.5% (3 of 8) of alveolar RMS (ARMS) cases; moreover, PIK3CA mutations were found in 25% (2 of 8) of ARMS specimens. The results demonstrate that tumor profiling in archival tissue samples is a useful tool for identifying diagnostic markers and potential therapeutic targets and suggests that these HRAS/ PIK3CA mutations play a critical role in the genesis of RMS.

• Restorative Dentistry and Endodontics
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• 제14권2호
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• pp.49-64
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• 1989

The purpose of this study was to evaluate the effect of several materials on the healing process of apical wound. Sixteen mandibular premolars obtained from 4 healthy dogs were used for this study. Under general anesthesia, the pulpal chamber of each tooth was opened and the pulps were extirpated. The root canals were then instrumented with H-file and irrigated with physiologic saline solution ; the apices were purposely perforated and enlarged with the engine K-reamer. In the experimental groups, apical wounds were filled with one of calcium hydroxide, hydroxylapatite, and tricalcium phosphate materials, mixture of each materials and physiologic saline solution, with a lentulo spiral. In the control group, apical wounds were not filled with any material. All the root canals were filled by the lateral condensation technique with gutta-percha cone and ZOE sealer. The access opening of all the teeth were closed with amalgam. On the 10, 20, 40 and 60th day after experiment, experimental animals were sacrificed. Segments of jaws, each containing one tooth, were fixed in 10% formalin solution and decalcified in Plank-Rychlo solution. The specimens were embedded in paraffin and serially sectioned to an average thickness of $6{\mu}m$. The sections were stained by hematoxylin-eosin and Masson's trichrome stain method and examined under light microscope. The results were as follows : 1. In the experimental groups, the new bone formations were observed in apical wounds. 2. Fourty days later, apical wounds were healed by granulation tissue in the experimental groups, but were not healed by granulation tissue in the control group, and the healing process of experimental groups were more rapid than that of control group. 3. Sixty days later, chronic inflammation disappeared in the experimental groups, and the materials used showed biologic affinity to the periapical tissue. 4. In all the groups, the resorption of cementum appeared on the 10th and 20th day after experiment, and the deposition of cementum appeared on the 40th and 60th day after experiment, especially showing narrowness of apical foramen due to newly formed cementum in calcium hydroxide group. 5. Calcum hydroxide and tricalcium phosphate particles were gradually resolved, but hydroxylapatite particles were not resolved through the experimental period.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권6호
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• pp.421-433
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• 2002

In order to elucidate the pathogenesis of cleft lip and palate, first of all, it is necessary to understand the developmental mechanisms of growth factors and extracellular matrix proteins in the tissues of cleft lip and palate. We have performed immunohistochemical studies on human cleft lip and palate tissues to elucidate the pathogenetic implications of cleft lip and palate. 16 specimens from postnatal human cleft lip and palate subjects and 17 specimens from autopsy of prenatal human cleft lip and palate were fixed in 10% buffered formalin, embedded in paraffin. The sections were routinely stained by hematoxylin and eosin, also stained by PAS, and followed by immunohistochemical stainings using the antiseras of growth factors and extracellular matrix proteins such as PCNA, S-100, c-erb-B2, MMP-3, MMP-10, HSP-70, transglutaninase-C, E-cadherin, VEGF, vWF. Both the prenatal and postnatal specimens of cleft lip and palate showed dysplastic proliferation of the basal cell layer, increased infiltration of melanocytes into mucosal epithelium, sebaceous gland hyperplasia ingrowing into the muscular tissue of lip and palate, and fatty infiltration into the submucosal deep connective tissue. The strong reactions of MMP-3 and HSP-70 were detected in the tissues of cleft lip and palate, especially increased in degenerating muscle bundles, while the immunostainings of PCNA and c-erb-B2 were weakly positive in the tissues of cleft lip and palate. These data suggest that the retrogressive tissue degeneration around the cleft areas persistently exist during the prenatal and postnatal period after cleft formation, and the sebaceous gland hyperplasia and fatty infiltration with the intense expression of MMP-3 and HSP-70 is closely related to the muscular degeneration around the cleft area.

• Applied Microscopy
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• 제15권2호
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• pp.98-110
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• 1985

This study was performed to clarify the morphological structures of the epithelia of the renal papilla, renal pelvis and ureter of the sheep (Ovis aries L.) through the light and scanning electron microscopes, Tissue specimens were taken from the renal papilla (common renal papilla and peripelvic column) and the renal pelvis (pelvis proper and pelvic pouch) of the kidney and the ureter. For the light microscopy, tissue blocks were fixed in 10 % neutral buffered formalin and embedded in paraffin wax, serially sectioned at a thickness of $6{\mu}m$. These sections were stained with hematoxylin-eosin and periodic acid-Schiff reaction. For the scanning electron microscopy, tissue blocks were prefixed in 1% glutaral-dehyde-1.5% paraformaldehyde solution and postfixed in 1% osmium tetroxide solution, dehydrated in graded alcohol, transferred to isoamyl acetate, and then dried by the critical point dryer (Polaron E 3000). These dried tissues were coated with gold and observed with a scanning electron microscope (JSM-35C), The results were as follows: The apex of the common renal papilla was lined with simple columnar epithelium having many microvilli on its luminal surface. Lateral portion of the papilla was lined with stratified epithelium $2{\sim}3$ layers thick, and its superficial cells were microvillar cells having many microvilli. The epithelium lining the peripelvic column was $1{\sim}2$ layers thick. The superficial layer was made of the microvillar cells, but a few microplica cells were appeared in the region near the pelvic pouch. The epithelium of the pelvic pouch was $1{\sim}2$ layered transitional type, and its superficial cells were microplica cells. The epithelia of the pelvis proper and ureter were $4{\sim}6$ layered transitional type, and their superficial cells were typical facet cells existing many round depressions and ridges of cell membranes of the luminal side.

• Tuberculosis and Respiratory Diseases
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• 제44권4호
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• pp.756-765
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• 1997

연구배경 : 종양세포 성장의 역동학적 성상은, 종양의 분열능을 직접적으로 반영하므로 종양의 예후 예견에 임상적 의의가 있다. PCNA는 DNA 합성에 관여하는 36KD의 핵단백질로서, 종양 조직에서 면역조직화학적 방법으로, 그 발현 정도를 계측하여 종양세포 증식정도의 반영으로서 이용하고 있다. 이에 저자들은 폐암의 조직에서 면역조직화화적 방법으로 PCNA의 세포의 분열능을 반영하는 다른 분자생물학적 인자 즉 S-주기비율과의 관계를 확인하고, PCNA의 발현정도에 따른 폐암환자들의 생존률을 검색하였다. 방 법 : 대상군은 원발성 비소세포 폐암으로 확진 받고, 외과적 절제술 후 파라핀에 보관된 57례의 폐암 조직을 사용하여 면역조직화학 염색법으로 PCNA의 암세포형, TNM 병기, 세포 분화도, S-주기 비율과 생존률과의 관계를 분석하였다. 결 과 : 본 병원에서 외과적 절제술을 시행한 57례중 남녀비는 43 : 14였고, 평균연령은 60세였다. PCNA의 발현 정도는 25%를 기준으로 하여 +, ++, +++, ++++로 구분하였다. PCNA의 발현은 편평상피암에서 선암보다 강하게 발현되었으며, TNM 병기에 따른 PCNA 발현의 차이는 없었다. 세포의 분화도에 따라서 미분화일수록 PCNA의 발현 정도는 높았으나, 유의한 차이는 없었다. S-주기 비율은 -17.9%, +18.3(${\pm}9.7$)%, ++18.6(${\pm}11.1$)%, +++19.6(${\pm}9.0$)%, ++++24.7(${\pm}8.2$)% 였으나 서로간의 유의성있는 차이는 없었다. PCNA의 발현 정도에 따른 2년 생존률과 중간 생존기간은 -50% 13.0개월, +75% 41.3개월, ++73% 33.6개월, +++67% 29.0개월, ++++25% 9개월로서 서로간에 유의한 차이를 보여주고 있다(P<0.05, Kaplan-Meier법, generalized Wilcox). 결 론 : 비소세포 폐암에서 PCNA의 발현정도는 편평상피암에서 선암보다 높았고, TNM 병기와 세포분화도에 따른 차이는 없었다. PCNA 발현정도에 따라 S-주기 비율은 증가한 듯하나 유의성은 없었다. PCNA 발현이 높을수록 2년 생존률과 중간 생존기간은 유의하게 불량하였다. 즉 결과적으로 PCNA 염색법은 비소세포폐암의 예후인자로서의 이용이 가능하리라 생각된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권1호
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• pp.1-12
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• 2006

Pleomorphic adenoma is the most common benign tumor in salivary glands, and occurred in frequency of 60% in parotid gland tumors, and 50% in submandibular gland tumors, and 25% in sublingual gland tumors. Histopathologically, pleomorphic adenoma is composed of epithelial cells and mesenchymal tissues, and called 'mixed tumor' because of morphological divergency. The cell structures of luminal area are composed of polyhedral and cuboidal secretory epithelial cells and modified myoepithelial cells around it, and mesenchymal tissue is composed of some myoepithelial cells and stromal tissue. In stromal tissue, myxoid change, chondroid change, or hyalinization can be seen even if bone tissue. In many studies, tumor cells of pleomorphic adenoma containing modified myoepithelial cell participate in synthesis of glycosaminoglycans. In this study, tissue sample of pleomorphic adenoma of human salivary gland were obtained from 20 surgical specimens, and all specimens were routinely fixed in 10% formalin and embedded. Serial 4-8${\mu}m$ thick sections were cut from paraffin blocks. The histopathologic evaluation was done with light microscopy. And, with immunohistochemical staining, characteristics of glycosaminoglycan were observed. And, for biochemical analysis of glycosaminoglycan, isolation of crude glycosaminoglycan from tumor tissue and immuno-blot analysis were carried out. With transmission electromicroscopy, tumor cells and biologic behavior of pleomorphic adenoma were observed with distribution and expression of glycosaminoglycan in tumor cells, The results were obtained as follows: 1. In immunohistochemical study, chondroitin 4-sulfate is highly postively stained in myxoid stromal tissue, and chondroitin 6-sulfate is highly positively stained in chondroid mesenchymal tissue, both glycosaminoglycans are positively stained in non-luminal cell of ductal area. 2. Dermatan sulfate and keratan sulfate is positively stained in periductal non-luminal tumor cells. 3. In immunohistochemical study, heparan sulfate is weakly stained in luminal cells and non-luminal cells around duct, and chondroid mesenchymal tissue. 4. In transmission electromicroscopic view, the tumor cells are composed of modified myoepithelial cells, and contain many microfilaments and well developed rough endoplasmic reticulum. 5. In Immuno-Blot analysis, the expression of glycosaminoglycans is expressed mostly in chondroitin 6-sulfate and chondroitin 4-sulfate. From the results obtained in this study, tumor cells of pleomorphic adenoma are composed of modified myoepithelial cells, and glycosaminoglycans of chondroitin 4-sulfate and chondroitin 6-sulfate mostly participate in the development of pleomorphic adenoma, but dermatan sulfate, keratan sulfate and heparan sulfate glycosaminoglycans were expressed variably.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4739-4743
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• 2012

Background: Predictive biomarkers for lung cancer recurrence after curative tumor resection remain unclear. This study set out to assess the role of FoxM1 in the recurrence of non-small cell lung cancer. Methods: Immunohistochemistry for FoxM1 expression was performed on paraffin-embedded tumor tissues from 165 NSCLC patients. Association of FoxM1 expression with clinicopathological parameters and disease free survival were evaluated. Results: Our results indicated FoxM1 expression to be significantly associated with poorer tissue differentiation (P =0.03), higher TNM stage (P <0.01), lymph node metastasis (P <0.01), advanced tumor stage (P <0.01), and poorer disease free survival (P <0.01). Multivariable analysis showed that FoxM1 expression increased the hazard of recurrence (hazard ratio= 1.96, 95% CI, 1.04-3.17, P <0.05), indicating that FoxM1 is an independent and significant predictor of lung cancer recurrence. Conclusion: Therefore, FoxM1 is an independent risk factor for recurrence of NSCLC. Elevated FoxM1 expression could be used as an indicator of poor disease free survival.