To clarify the effect of cisplatin (CDDP) on proliferation and metastasis of rabbit VX2 tongue cancer, the cell proliferation factor (Ki-67) and the metastasis suppresser factor (nm23) were immunohistochemically investigated. <br>Three days after transplantation of VX2 cancer cells into the tongue in rabbits, physiological saline was administered to control groups (n=15), and 2 mg/kg and 4 mg/kg CDDP were administered to experimental groups I (n=15)and Ⅱ (n=15), respectively. The rabbits were sacrificed on 7 (n=5),14 (n=5), and 21(n=5}days after transplantation. The tongue, deep cervical lymph node, and lung were excised, the volumes of the tongue tumor and the deep cervical lymph node were measured. Tumor foci of the lung metastasis were macroscopically counted. The excised tissues were embedded in paraffin in the conventional method and serial sections of 4μm thickness were prepared. The sections were used for H-E staining and immunohistochemical stalnlng.<br>The Ki-67 labeling index of the tongue tumor increased in proportion to the tumor volume, and this index was significantly increased in proportion to tumors that metastasized to the deep cervical lymph node and the lung. The degree of nm23 expression in tongue tumors was decreased when the tumor was metastasized to the deep cervical lymph node and the lung, indicating a relationship between nm23 expression and tumor metastasis. As the tongue tumor grew, tumor cells metastasized to the deep cervical lymph node on the ipsilateral side with transplantation, then to the deep cervical lymph node on the contralateral side without transplantation. In experimental group I,metastasis to the deep cervical lymph node on the contralateral side was observed 21 days after transplantation, while in experimental group Ⅱ,metastasis was not yet found in the lymph node on the contralateral side. Metastasis to the lung occurred after metastasis to the deep cervical lymph node was observed. In experimental group I,lung metastasis was found in most of the rabbits 21 days after transplantation, while lung metastasis was not observed in experimental group Ⅱ.<br>Therefore, the proliferation of the VX2 tongue tumor in the rabbit was more strongly inhibited in experimental group Ⅱ than that in experimental group I,showing a high expression of cancer metastasis suppresser factor. These findings suggested that adrninistration of 4 mg/kg CDDP is effective in inhibiting metastasis because it delays metastasis to the deep cervical lymph node and the lung.

In the present study, the histogenesis and fate of the tectum cartilage appearing under the sagittal suture from day l6 of embryonic gestation to 15-day-old mice were examined histochemically and immunohistochemically. Cartilage tissues were persistent as alcian blue-positive cartilaginous aggregates under the posterior fontanel and sagittal suture from 17 days embryonic development until day 9 after birth. The size of these cartilage specimens reached a maximum at 6 days old and disappeared by 10 days old accompanying sutural ossification.

<br>Connective tissues, including the perichondrium of the tectum cartilages, and calvarial bone were immunopositive for type I collagen, whereas the cartilage matrix was a typical hyaline cartilage showing positive features for anti-type Ⅱ collagen antibody during the degeneration of the cartilage. The tectum cartilage at 6 days old was comprised of hypertrophic chondrocytes showing immunoreactivity for type X collagen, and it was confirmed by BrdU-incorporation that these hypertrophic cells had no mitotic capacity. Apoptotic cells were not detected in the hypertrophic chondrocytes during the degenerating stage at 9 days old. TRAP (tartrate-resistant acid phosphatase)-positive cells and macrophages showing resorption of cartilage matrix and cellular debris were not observed in the degenerating tectum cartilage. Immunoreactivity for MMP-1(matrix metalloproteinase), indicating the existence of collagenase, was visualized in the cartilage matrix<br>These results suggested that tectum cartilage retained features of hyaline cartilage, and its disappearance may be induced by matrix degradation with collagenase.

The importance of radiography in determining prognosis after dental implantation has been strongly urged in recent years. In this study, we examine the relationship between the fine shadow in radiograph images and the actual inner structure of the mandible by comparing a radiograph with computer graphic three dimensional reconstruction imagery (CG 3-D) created from serial two-dimensional tissue images.<br>

Initially, we used Particle Induced X-ray Emission analysis to distinguish bone mineral characteristics of each sample tissue. Our results indicate that while the specific bone mineral values differed between mandible samples, no difference was observed in values between buccal and lingual cortical bone and cancellous bone in individual samples. Since no differences in bone mineral deposition was found across the mandible, it was presumed that there is a close relationship between the bucco-lingual thickness of the mandible and radiographic density, rather than between the thickness of individual bone regions and image density.

<br>When we compared CG 3D images to the radiographs, we found that the nutrient canal in the lingual side of the cortical bone produced radiolucency in the radiograph while few nutrient canal resulted in increased radiopacity. The ratio of the nutrient canal in the lingual cortical bone correlated well with the radiographic density of the radiograph, especially in regions of radiopacity.

<br>Clinically, the fine shadow on mandibular radiographs has been attributed to the nutrient canal structure in the cortical bone on the lingual side. We think that it does not reflect differences in structure between the buccal cortical bone, cancellous bone, and the lingual cortical bone, From this study, it is clear that diagnosis should be made after multiple types of examination in addition to radiography.

The influence on the gingivae of female hormones is clinically known for its relation with pregnancy gingivitis, pregnancy tumor and desquamative gingival lesion. In the previous paper, the author investigated the influence on the gingivae of Pelanin depot^TM(PD) from the standpoint of wound healing using old beagle dogs. Consequently, the administration of PD brought about a loss of body weights, a rise of serum estradio1-17 β (E₂) values, and an obvious delay of wound healing in the gingivae.<br>The present study was conducted in order to investigate the relation of wound healing of the gingivae, age and sex to PD-administration using adult beagle dogs. <br>13 beagle dogs (7 males and 6 females of 2 to 5 years old) were used for this experiment, and the animals were divided into control (4 males) and experimental (3 males and 6 females) groups. The experiment was carried out using the same method as the previous one.<br>The following results were obtained from this experiment. <br>1. In the experimental group, the body weights of males showed a slight increase and the females showed a slight decrease, but the control group showed no changes. <br>2. Serum E₂ values of males and females after PD-administration in the experiment group were larger than the values before PD-administration, where as serum E₂ values in the control group were below the minimum quantity (10 pg/ml) that can be measured per pg uilt. <br>3.In the macroscopic observation of the experiment group, the duration until healing after gingivectomy was prolonged compared to that of the control group. <br>4.In the microscopic observation of the experiment group, a remarkable infiltration of inflammatory cells was found in the first week after gingivectomy, but was no found 4 th weeks after, although the control group displayed a disappearance of inflammatory cells at the 3 rd weeks after gingivectomy.<br>The present study suggests that the administration of female hormones causes a delay of wound healing in the gingivae.

The purpose of this study was to clarify the effects and mechanisms of midazolam on changes in contraction of the smooth muscle induced by a voltage dependent Ca²⁺ channel stimulator,50mM KCI, and receptor activated Ca²⁺ channel agonist, serotonin(5-HT), in bovine middle cerebral arteries.<br>

The isometric tension and intracellular Ca²⁺consentration ([Ca²⁺]i)were measured simultaneously by the fura-2 microfluorometric methods, and I tried to deduce from their relationship the mechanism of midazolam on contraction.<br>

The results obtained were as follows:<br>

(1) KCl and 5-HT developed the degree of contraction and[Ca²⁺]iin a concentration-dependent manner. <br>

(2) Midazolam depressed the degree of contraction and[Ca²⁺]iinduced by KCI and 5-HT in a concerltration-dependent manner. <br>

(3) Midazolam had no effect on the tension/[Ca²⁺]i　of KCI and 5-HT.<br>

(4) Midazolam had no effect on transient increase of tension and [Ca²⁺]i induced by 5-HT and caffeine in Ca²⁺-free physiological salt solution (Ca²⁺-free PSS).<br>

(5) Flumazenil, a specific central-type antagonist of benzodiazepines, and PK-11195, a specific periphera1-type antagonist of benzodiazepines, had no effect on the depression of tension and [Ca²⁺]i by midazolam.<br>

These results suggest the conclusion as follows,<br>

Midazolam does not depress the contraction of bovine middle cerbral arteries iduced by KCI and 5-HT in the manner of inhbition of Ca²⁺ release from sarcoplasmic reticulum and sensitivity of contractile elements to Ca²⁺ but does in the manner of inhbition of Ca²⁺ influx into smooth muscle cell from outside. Midazolam may not inhibit the contraction of smooth muscle through the central nor peripheral benzodiazepine receptors.

Apoptosis is different from necrosis of the shell in terrns of the morphology of its process. It has been shown that apoptosis is induced by many anticancer drugs. Recently, there are a lot of reports about the relationship between the cell cycle and apoptosis in tumor inhibition, but the relationship between the cell cycle and the morphology alteration of apoptosis <i>in vivo</i> have not been demonstrated. This study shows the relationship between the cell cycle and apoptosis of the murine squamous cell carcinoma cells with a Laser Scanning Cytometer (LSC). As a result, There was a significant correlation between the incidence of apoptosis and tumor inhibition by cisplatin in the murine squamous cell carcinoma cells. And the incidence of apoptosis increased in a dose-dependent manner. There was a trend toward higher incidence of the Inurine squamous cell carcinoma cells in the G2+M, G0+G1 phase in cisplatin adrninistration group than in control group. In each phase of the cell cycle, the sequential change of the Apoptotic Index (AI) was not recognized.<br>In this study, it was confirmed that apoptosis was induced by cisplatin in the murine squamous cell carcinoma cells. Furthermore, the relatiorlship between the cell cycle regulation mechanism and AI was suggested.

Astatistical analysis of the autopsy cases of malignant tongue neoplasms from the Annual of Pathological Autopsy Cases in Japan during a 5-year-period (1992-1996) was reported for the purpose of clarifying the nature of the tongue cancer. <br>During this period 362 autopsy cases (252 males, 110 females) of malignant tongue neoplasms were reported. The autopsy rate has recently decreased and the mean autopsy rate for these 5 years was 7.8% among the patients who died of malignant tongue neoplasms which were reported in the Vital Statistics of Japan (Volume 3). <br>The autopsy cases of malignant tongue neoplasms occurred in the seventh decade(106 cases, 29、2%),the eigth decade(99 cases,27.3%), the sixth decade(65 cases,18.0%), and in individuals more than 90 year old(6cases). It did not occur in the second decade. Histologically, almost all cases showed squamous cell carcinoma. The cancer arose in the lateral border of the tongue (31 cases, 55.4%), in the root of the tongue (30.4%), and in the frontal tongue (7.1%). <br>Mean age for the autopsy cases (only the cases of squamous cell carcinoma excepting for multiple primary cancers were analyzed), were 64.2±11.1 (1992),64.3±12.7 (1993),63.2±13.3 (1994),65.9±12.7 (1995),and 68.2±14.2 year-old(in the year 1996), respectively. Mean age gradually increased over the 5 years, and it was higher in females than in males in each year. <br>Multiple primary cancers, affecting both the tongue and other organs, were found in 122 cases (double cancer:90 cases, threefold cancer:22 cases, fourfold cancer:7 cases, fivefold cancer:3 cases). Multiple primary cancer with the tongue cancer occupied 28.2%of all primary cancers in 1992,35.4% in l993,31.6%in 1994,42.9% in 1995, and 36.5% in 1996, respectively. Double cancers with the tongue cancer progressed in the esophagus, lung, stomach, and colon. Metastasis to other organs was found in the lung,1iver, pleura, adrenal gland, kidney, and thyroid gland. Metastasis to the lymph node was found in the cervical, pulmonary hilar, paraesophagea1, and paratrachial nodes. <br>The most common causes of death not associated with malignant neoplasms of the tongue were the pulmonary infection, disrupture of the blood vessels, bleeding from the cancer, asphyxia, ulcers or bleeding of the digestive organs, and the sepsis.

We studied the postoperative stability of mandible following the combination of intraoral inverted L ramus and sagittal splitting ramus osteotolny (I-S) in mandibular asymmetry patients. Postero-anterior and lateral cephalometric radiograms of the I-S group (n=7) and of the bilateral sagittal splitting osteotomy (SSRO) group (n=6) were analyzed in the following items: 1) The ratio of the deviation side area to the non deviation side area in the lower mandible, 2)The ratio of the deviated side distance to the non deviated side distance of the Go-midline, 3) Ramus angle, 4) Facial angle, 5) SNB, 6) Mandibular angle etc. Each value of the I-S group was compared with those of the SSRO group. There were no significant differences between the two groups, and little change during the I2 months postoperative period, Consequentry, I-S stabilizes the mandible Postoperatively as well as SSRO does.

Retinoic acid is a signaling molecule involved in the regulation of the growth and differentiation of various normal and transformed epithelial cells. Retinoids exert their action through two families of nuclear receptors:retinoic acid receptors (RARs) and retinoid X receptors (RXRs). There are three types of RAR α, β, γ for all<i> trans</i>-retinoic acid (<i>at</i>-RA) in mammals. To better understand the effect of <i>at</i>-RA on proliferation and RAR-expression of a human salivary gland adenocarcinoma cells (HSG-S 8), growth, DNA histograms, Northern blot of <i>c-myc</i> mRNA,TUNEL-positive apoptotic cells, immunohistochemisry of keratins and the expression level of RAR by RT-PCR were analyzed in HSG-S 8 cells cultured with <i>at</i>-RA. A<i>t</i>-RA decreased the expression of c-<i>myc</i> mRNA and induced G1 arrest, resulting in a dose-dependent inhibition of cellular growth of HSG-S8. <br>However, HSG-S 8 did not undergo apoptosis, since the frequencies of TUNEL-positive cells in the group of the culture with <i>at</i>-RA and in those of the group of the culture without <i>at</i>-RA were almost the same。 RARα-selective agonist (Ro40-6055) inhibited the incorporation of [³H]-thymidine into HSG-S 8 in a similar dose-dependent manner to that of <i>at</i>-RA. Immunoreaction of molecular keratin peptides with PKK-1monoclonal antibody decreased in <i>at</i>-RA-treated cells. On the other hand immunoreaction with 68kD cytokeratin monoclonal antibody increased in HSG-S8. Namely, <i>at</i>-RA induced squamous metaplasia of HSG-S8. These results indicated that <i>at</i>-RA inhibited the growth of HSG-S 8 because of down-regulation of cc and subsequent G l arrest, but did not induce apoptosis of cells. These actions of <i>at</i>-RA on HSG-S 8 cells seemed to be mediated by RARα.

The relationship between proliferative activity and the expression of cell-cell adhesion molecules in tumor cells in ameloblastoma was immunohistochemically examined according to the localization of proliferating cell nuclear antigen (PCNA), E-cadherin and β-catenin. <br>Specimens were collected from 29 patients for 22 years from 1978 to1999. The specimens were extracted at biopsy or surgical operations. They were histopathologically diagnosed as 26 benign ameloblastomas including three unicystic ameloblastomas, 11 plexiform patterns, 12 follicular patterns, and three malignant ameloblastomas. These specimens embedded in paraffin were cut into sections of 4μm thickness, and were stained with conventional hematoxylin and eosin stain. They were further immunostained with anti-PCNA, E-cadherin and β-catenin monoclonal antibodies. The labeling index (LI) was calculated as the frequency PCNA-positive cells in the tissue of each ameloblastoma.<br>The monoclonal antibody against PCNA stained the nuclei of tumor cells and sometimes those of stromal cells. A different localization of PCNA-positive cells was found among the three types of benign ameloblastomas and malignant ameloblastoma. PCNA-positive cells were present in the peripheral and the central areas of the tumor nests in unicystic ameloblastomas and plexiform patterns. On the other hand, they existed mainly in the peripheral layer of the tumor nests in follicular patterns and were scattered throughout the atypical-cell-abundant portions in the malignant ameloblastoma. E-cadherin and β-catenin showed similar localization and appeared in intercellular bridges in the prickle cell layer, but not in the basal cell layer in normal oral mucosa. In plexiform patterns, E-cadherin and β-catenin were clearly stained in cellular membranes and in the cytoplasm of the basal portions of the marginal tumor cells in the tumor nest, and less stained in follicular patterns than plexiform ones. Althogh they were detected in the portions showing either plexiform or follicular features in malignant ameloblastoma, E-cadherin and β-catenin were scarcely immunostained in atypical-cell-abundant portions. The PCNA-LIs increased in the order of the plexiform pattern, follicular pattern, and malignant ameloblastoma, whereas the staining of both the E-cadherin and β-catenin tended to reduce in the same order.<br>These results indicated that the proliferative activity and the degree of expression of cell-cell adhesion molecules were inversely correlated in ameloblastomas.

We report on a cystic lymphangioma in the left cervical area in the submandibular reslon. <br>A 17-year-old female developed swelling in the left submandibular area at the age of about 10 years, and repeatedly underwent puncture aspiration of the content fluid at a local hospital. However, the swelling gradually increased, extending to the left cervical region. The lesion showed wide extension, MRI and CT preoperative imaging diagnosis of was useful for confirming the extension area of the cyst. Though the lesion was surgically extirpationed as much as possible, complete extirpation was difficult. <br>Her course has been good a postoperative period of four years, without any recurrence.

Behcet’s disease is a multisystem inflammatory disorder that includes mucocutaneous, ocular and genital involvement. It’s etiology is unknown, and the treatment has not been established completely. We have described our experience with a 64-year-old woman with complete type Behcet’s disease. The patient was given Unsei-in (TJ-57) and Syakuyaku-kanzoutou (TJ-68). These crude drugs were effective in the treatment of Behcet’s disease.