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.
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.
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.
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
/
v.28
no.6
/
pp.421-433
/
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.
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.
The purpose of this study was the observe the toxic effects of root canal sealers in 108 white rats. Experimental animals were divided into control and experimental groups. Theree representative types of materials, such as AH26, Z.O.E. and F.R. were used in this study. Cavities were prepared on the left mandibular area of 108 white rats. Three different sealers were placed in as experiment and bone cavities were left without filling as control. The experimental animals were sacrificed by cervical dislocation at the intervals of 1, 3, 7, 14, 28 and 49 days after filling. Each specimen was fixed with 10% neutral formalin solution, decalcified with 5% nitric acid, embedded in paraffin and sectioned 5-7${\mu}$. in thickness. The paraffin sections stained with Hematoxylin - Eosin were observed through the ordinary light microscope. The results were as follows; 1. Slight toxic effect to surrounding tissue were found in every experimental specimen. 2. AH26 showed the highest inflammatory response, and F.R. showed the lowest inflammatory response which subsided and replaced by fibrosis at 4 weeks after filling. 3. The cavity filled materials, such as implanted root canal sealers, blood clots and necrotic tissue, showed a tendency to be absorbed gradually proportioned to the experimental periods. A small amount of cavity filled materials were observed in the bone cavities after 4 weeks. 4. Fibroblastic proliferation began to produce fibrous capsule around the bone cavity in 2 weeks after filling. Fibrosis was prominent at 4 weeks after filling. 5. Osteoblastic activity of surrounding bone was observed at first in 2 weeks after filling and prominent in 4 weeks after filling. Osteoblastic activity showed an increasing effect as the time prolonged. 6. Surrounding tissue of the bone cavities showed the features of tissue destruction and had very severe inflammatory response at an initial stage. Above-mentioned appeared to be recovered gradually proportioned to the experimental periods.
Considerable attention has been given to the accuracy of HER-2 testing and the correlation between the results of different testing methods. This interest reflects the growing importance of HER-2 status in the management of patients with breast cancer. In this study the detection of HER-2 gene and centromere 17 status was evaluated using dual-colour primed in situ labelling (PRINS) in comparison with fluorescence in situ hybridization (FISH). These two methods were evaluated on a series of 27 formalin fixed paraffin embedded breast carcinoma tumours, previously tested for protein overexpression by HercepTest (grouped into Hercep 1+/0, 2+ and 3+). HER-2 gene amplification (ratio${\geq}2.2$) by PRINS was found in 3:3, 6:21 and 0:3 in IHC 3+, 2+ and 1+/0 cases, respectively. Comparing FISH and IHC (immunohistochemistry), showed the same results as for PRINS and IHC. Chromosome 17 aneusomy was found in 10 of 21 IHC 2+ cases (47.6%), of which 1 (10%) showed hypodisomy (chromosome 17 copy number per cell${\leq}1.75$), 7 (70%) showed low polysomy (chromosome 17 copy number per cell=2.26 - 3.75) and 2 (20%) showed high polysomy (chromosome 17 copy number per cell ${\geq}3.76$). The overall concordance of detection of HER-2 gene amplification by FISH and PRINS was 100% (27:27). Furthermore, both the level of HER-2 amplification and copy number of CEN17 analysis results correlated well between the two methods. In conclusion, PRINS is a reliable, reproducible technique and in our opinion can be used as an additional test to determine HER-2 status in breast tumours.
Objective : The aim of this study was to evaluate the microanatomy and histological features of the central myelin in the root exit zone of facial nerve. Methods : Forty facial nerves with brain stem were obtained from 20 formalin fixed cadavers. Among them 17 facial nerves were ruined during preparation and 23 root entry zone (REZ) of facial nerves could be examined. The length of medial REZ, from detach point of facial nerve at the brain stem to transitional area, and the thickness of glial membrane of central myelin was measured. We cut brain stem along the facial nerve and made a tissue block of facial nerve REZ. Each tissue block was embedded with paraffin and serially sectioned. Slices were stained with hematoxylin and eosin (H&E), periodic acid-Schiff, and glial fibrillary acid protein. Microscopy was used to measure the extent of central myelin and thickness of outer glial membrane of central myelin. Thickness of glial membrane was examined at two different points, the thickest area of proximal and distal REZ. Results : Special stain with PAS and GFAP could be differentiated the central and peripheral myelin of facial nerve. The length of medial REZ was mean 2.6 mm (1.6-3.5 mm). The glial limiting membrane of brain stem is continued to the end of central myelin. We called it glial sheath of REZ. The thickness of glial sheath was mean $66.5{\mu}m(40-110{\mu}m$) at proximal REZ and $7.4{\mu}m(5-10{\mu}m$) at distal REZ. Conclusion : Medial REZ of facial nerve is mean 2.6 mm in length and covered by glial sheath continued from glial limiting membrane of brain stem. Glial sheath of central myelin tends to become thin toward transitional zone.
Recently, available interests concerning the biologic significance of the extracellular matrix and proliferating cells associated with periodontal disease has been increased. The distribution or expression of cellular proliferation by PCNA, macrophage detection by ${\alpha}$-l-antichymotrypsin, fibronectin playing a important role in host defence mechanisms indirectly, and transglutaminase that cross linked to fibronectin and stimulate fibrin stabilization were studied in inflammed and healthy gingiva. The excised tissue samples were fixed neutral formalin for 24 hours, embedded with paraffin, sectioned at 4-61lffi in thickness, and immunohistochemically processed by LSAB method. The positive reaction to PCNA was localized in the suprabasal and basal layer of inflammed gingiva and an increasing reactivity was observed than healthy gingiva. ${\alpha}$-I-antichymotrypsin positive cells were localized in the basal layer of inflammed gingiva, and there was no or rare positive cells in healthy gingiva. The positive reaction to fibronectin in inflammed gingiva was more than healthy gingiva,"and shown in the connective tissue subjacent to basement membrane of epithelium and in the periphery of the collagen fiber bundles. The positive cells by transglutaminase in inflammed gingiva were noted in suprabasal, spinous, and keratin layer of epithelium, and slightly increased in the capillaries of connective tissues. But the results of this study demonstrated in vitro reaction. Therefore, the role of PCNA,${\alpha}$-l-antichyrnotrypsin, transglutaminase, fibronectin and coefficient with other growth factor and extracellular matrix were further investigated in vivo.
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