Variant muscle fibers connecting the orbicularis oculi to the orbicularis oris: case report / 대한해부학회지

The orbicularis oculi (OOc) is a sphincteric muscle of the eyelids, whereas contraction of the orbicularis oris (OOr), another sphincteric muscle, causes narrowing of the lips. Facial muscle fibers normally blend with adjacent muscles. However, muscle fibers connecting the various facial muscles that have different actions and that are located at distant sites, such as the OOc and the OOr have been rarely reported. Herein, we report a rare case of connecting fibers between the inferior margin of the OOc and the OOr. These connecting fibers were blended with the OOr between the inserting fibers of the levator labii superioris and levator anguli oris. Contraction of such variant muscles might affect typical facial expressions.

Comparison of immunohistochemical analysis on sinus augmentation using demineralized tooth graft and bovine bone

Objectives@#The purpose of this animal research was to compare bone regeneration in augmented rabbit maxillary sinuses treated with demineralized particulate human-tooth graft and anorganic bovine bone by immunohistochemical analysis. @*Materials and Methods@#Piezoelectric bilateral sinus augmentation was performed in eight adult rabbits. In the control group, anorganic bovine was grafted in the maxillary sinus following elevation of the sinus membrane. In the experimental group, demineralized human particulate tooth bone was grafted in the sinus. Bone regeneration in augmented sinuses was evaluated by immunohistochemical analysis using various markers of osteoprogenitor cells. @*Results@#The number of bromodeoxyuridine-labeled cells was significantly higher in the experimental group than in the control group at eight weeks. The immunoreactivity of proliferating-cell nuclear antigen was increased slightly in the experimental group relative to the control group at eight weeks. Other bone markers were expressed equally in the two groups. @*Conclusion@#In the rabbit maxillary sinus, higher osteoinduction was correlated with demineralized human particulate tooth bone grafting than with anorganic bovine grafting.

Comparison of immunohistochemical analysis on sinus augmentation using demineralized tooth graft and bovine bone

Objectives@#The purpose of this animal research was to compare bone regeneration in augmented rabbit maxillary sinuses treated with demineralized particulate human-tooth graft and anorganic bovine bone by immunohistochemical analysis. @*Materials and Methods@#Piezoelectric bilateral sinus augmentation was performed in eight adult rabbits. In the control group, anorganic bovine was grafted in the maxillary sinus following elevation of the sinus membrane. In the experimental group, demineralized human particulate tooth bone was grafted in the sinus. Bone regeneration in augmented sinuses was evaluated by immunohistochemical analysis using various markers of osteoprogenitor cells. @*Results@#The number of bromodeoxyuridine-labeled cells was significantly higher in the experimental group than in the control group at eight weeks. The immunoreactivity of proliferating-cell nuclear antigen was increased slightly in the experimental group relative to the control group at eight weeks. Other bone markers were expressed equally in the two groups. @*Conclusion@#In the rabbit maxillary sinus, higher osteoinduction was correlated with demineralized human particulate tooth bone grafting than with anorganic bovine grafting.

Comparison of Regeneration Effects of Direct and Alternating Microcurrent Therapy on Atrophied Calf Muscle in a Rabbit

Objective@#We compared the regenerative effects of microcurrent therapy (MT) according to the type of electric current, which were direct current microcurrent therapy (DCMT) and alternating current microcurrent therapy (ACMT) on atrophied calf muscle in cast-immobilized rabbit. @*Methods@#Rabbits were allocated into control group (sham MT), ACMT group, and DCMT group.Before starting treatment, right gastrocnemius (GCM) muscle was immobilized by cast for 2 weeks. Compound muscle action potential of tibial nerve in nerve conduction study, circumference of calf muscle using a ruler, and thickness of medial and lateral GCM muscle measured by ultrasound, cross sectional area (CSA), and proliferating cell nuclear antigen (PCNA) ratios (%) of muscle fibers were measured on the immunohistochemical analysis. @*Results@#The mean atrophic changes (%) in right medial and lateral GCM muscle thickness, right calf circumference, and amplitude of CMAP of the right tibial nerve in ACMT group and DCMT group were significantly lower than those in control group, respectively (p＜0.05). The mean CSA (μm 2 ) of type I and type II and PCNA ratios (%) of medial and lateral GCM muscle fibers in ACMT group and DCMT group were significantly greater than those in control group, respectively (p＜0.05). There were no significant differences between the ACMT group and DCMT group at all parameters. @*Conclusion@#This study demonstrated that ACMT and DCMT showed better regeneration effect than sham MT. Microcurrent may be effective in regeneration of atrophied muscle regardless of the type of current.

Comparison of Regeneration Effects of Direct and Alternating Microcurrent Therapy on Atrophied Calf Muscle in a Rabbit

Objective@#We compared the regenerative effects of microcurrent therapy (MT) according to the type of electric current, which were direct current microcurrent therapy (DCMT) and alternating current microcurrent therapy (ACMT) on atrophied calf muscle in cast-immobilized rabbit. @*Methods@#Rabbits were allocated into control group (sham MT), ACMT group, and DCMT group.Before starting treatment, right gastrocnemius (GCM) muscle was immobilized by cast for 2 weeks. Compound muscle action potential of tibial nerve in nerve conduction study, circumference of calf muscle using a ruler, and thickness of medial and lateral GCM muscle measured by ultrasound, cross sectional area (CSA), and proliferating cell nuclear antigen (PCNA) ratios (%) of muscle fibers were measured on the immunohistochemical analysis. @*Results@#The mean atrophic changes (%) in right medial and lateral GCM muscle thickness, right calf circumference, and amplitude of CMAP of the right tibial nerve in ACMT group and DCMT group were significantly lower than those in control group, respectively (p＜0.05). The mean CSA (μm 2 ) of type I and type II and PCNA ratios (%) of medial and lateral GCM muscle fibers in ACMT group and DCMT group were significantly greater than those in control group, respectively (p＜0.05). There were no significant differences between the ACMT group and DCMT group at all parameters. @*Conclusion@#This study demonstrated that ACMT and DCMT showed better regeneration effect than sham MT. Microcurrent may be effective in regeneration of atrophied muscle regardless of the type of current.

Histomorphometric study of rabbit's maxillary sinus augmentation with various graft materials / 대한해부학회지

The purpose of this animal study is to evaluate, by histomorphometric analysis, bone regeneration in rabbit's maxillary sinuses with blood clots alone, Bio-Oss, β-tricalcium phosphate (β-TCP), and demineralized tooth dentin (DTD) grafting. Bilateral sinus augmentation procedures were performed in 18 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the group 1, blood clots were filled; group 2, anorganic bovine graft (Bio-Oss) was grafted; group 3, β-TCP was grafted; group 4, DTD was grafted, and covered by replaceable bony windows. Animals were sacrificed at 2, 4, and 8 weeks after surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin and eosin and Masson's trichrome stains. Histologically, new bone formation was revealed along the elevated sinus membrane and all graft materials. The new bone area of the group 2 was significantly greater than the group 1, and of the group 3 was significantly greater than the group 2, and of the group 4 was significantly greater than the group 3 at 8 weeks with P < 0.05. The bone marrow area of group 1 was significantly greater than other groups at 8 weeks. The DTD area was significantly lesser than Bio-Oss or β-TCP particles area at 8 weeks. This present study suggests that DTD can be effective graft materials for bone regeneration of the maxillary sinus augmentation.

The Effects of Moxifloxacin in Endotoxin-induced Acute Lung Injury / 대한흉부외과학회지

BACKGROUND: The pathophysiology of acute respiratory distress syndrome with sepsis is acute lung injury (ALI) that's' caused by endotoxin (LPS). We evaluate effects of moxifloxacin on LPS-induced ALI in a rat model. MATERIAL AND METHOD: The rats were divided into 3 groups as the control group (C), the LPS insult group (L), and the LPS+moxifloxacin treated group (L-M). ALI was induced by endotracheal instillation of E.coli LPS, then moxifloxacin was given in 30 minutes. Five hours later, we checked the lung weight/body weight ratio(the L/BW ratio), the protein & neutrophils in the bronchoalveolar lavage fluid (BALF), the myeloperoxidase (MPO) activity & the malondialdehyde (MDA) content, the expressions of cytosolic and secretory phospholipase A2 (c, sPLA2), and the morphology of the lung with using a light microscope. RESULT: The L/BW ratio, the protein content and the neutrophil count in the BALF, and the MPO activity and the MDA content in lung were significantly increased in group L compared to group C, and these factors were markedly decreased in group L-M compare to group L. The cPLA2 expression and the sPLA2 expression were increased in group L and the cPLA2 expression was decreased in group L-M. Yet the sPLA2 expression was not changed in group L-M. Morphologically, many inflammatory findings were observed in group L, but not in group L-M. CONCLUSION: Many of the inflammatory changes of ALI that were caused by LPS insult were ameliorated by moxifloxacin treatment.

The Properties of Glycoconjugates in the Rat Nasal Mucosa following Inhalation of Formaldehyde / 대한해부학회지

The present study was carried out to investigate the glycoconjugate properties of the nasal mucosa in the rat after inhalation of formaldehyde. Sprague-Dawley male rats were inhalated 30 ppm formaldehyde for 3 times with 3 hours exposure. The olfactory and respiratory mucosa in the nasal mucosa were taken from the animals on 3, 6,9 days and 2, 3, 4, 5 weeks after inhalation of formaldehyde. The properties of glycoconjugate of the olfactory and respiratory mucosa were investigated using nine biotinylated lectins (PSA, UEA I, PHA-L, BSL I, PNA, MAL I, DBA, BSL II or sWGA). In experimental groups, the degenerative changes of the olfactory epithelium were observed until 3 weeks after inhalation of formaldehyde, but the respiratory epithelium was no change. In control group, the olfactory cells in the olfactory epithelium reacted with PSA, UEA I, PNA, DBA, BSL II, sWGA, and the supporting cells reacted with PSA, PHA-L, PNA, MAL I, DBA, BSL II, sWGA, and Bowman's glands reacted with all the lectins. In experimental groups, the olfactory cells reacted with UEA I, DBA, and the supporting cells reacted with PHA-L, MAL I, DBA, UEA I, and the positive reaction of Bowman's glands was increased. In control group, the goblet cells in the respiratory epithelium reacted with UEA I, MAL I, and the ciliated columnar cells reacted with PSA, UEA I, PHA-L, BSL I, DBA, BSL II, sWGA, and the septal nasal glands reacted with all the lectins except UEA I. In experimental groups, the goblet cells reacted with UEA I, MAL I and PNA. Conclusively, the olfactory mucosa was shown a lot of changes in the properties of glycoconjugates following inhalation of formaldehyde, but respiratory mucosa was shown feeble change. These results suggest that there were different sugar residues of glycoconjugate in the olfactory and respiratory mucosa following inhalation of formaldehyde, respectively.

Adaptive Response and Apoptosis Induced by UV Irradiation in Human Keratinocytes and Squamous Cell Carcinomas / 대한해부학회지

The present study was investigated that the effects of suitable ultraviolet (UV) irradiation to the cell viability, apoptosis and expression of poly(ADP-ribose) polymerase (PARP) in HaCaT and KUMA3 cell lines that are skin keratinocyte and squamous cell carcinoma, respectively. The cell viability of UV irradiation dose appeared high in HaCaT cell line, and the percentage of apoptotic cells appeared higher in KUMA3 cell line. The cell viability in HaCaT cell line pretreated with 5 J/m(2) UV and subsequently treated with 50 J/m(2) UV was higher than that only treated with 50 J/m(2) UV. On the other hand, the percentage of apoptotic cells in KUMA3 cell line only treated with 50 J/m(2) UV was higher than that pretreated with 5 J/m(2) UV and subsequently treated with 50 J/m(2) UV. Immunocytochemical stain using PARP antibody, the most cells show positive reaction of UV irradiation in HaCaT cell line, but some negative reaction cells observe from treated with 50 J/m(2) UV in KUMA3 cell line. The expression level of PARP by western blot analysis, concomitant with generation of the 85 kDa fragment in HaCaT cell line from treated with 50 J/m(2) UV though the 116 kDa band was still retained. On the other hand, the 85 kDa fragment appeared in KUMA3 cell line from treated with 50 J/m(2) UV but the intact PARP of 116 kDa disappears in more doses. These results present that suitable UV irradiation can be effective to apoptosis in squamous cell carcinoma.

Effects of bears' bile on the expression of tumor cell p53 protein / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effects of bears' bile on tumor cell p53 protein expression with different gene properties.</p><p><b>METHODS</b>The effects of bears' bile on the expression of p53 protein in 6 cancer cell strains were determined by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Results Western blot analysis showed that the expression of p53 protein in HaCaT, KUMA3, KUMA4 and KUMA6 cell strains with gene mutation were increased, but no change was found in HCT116 and KUMA5 cell strains without gene mutation. There was no quantitative change in p53 mRNA in all cell strains by analysis of p53 mRNA with</p><p><b>CONCLUSION</b>The effects of bears' bile on p53 protein expression in cancer cell strains RT-PCR analysis system. could be different based on p53 gene properties,i. e. ,bears' bile only affect p53 protein of mutation type.</p>

Morphological Study for Osteoinduction by Human Fibroblasts Transduced with rhBMP-7 Adenovirus / 대한해부학회지

To evaluate availability of the BMP-7 adenovirus (AdBMP-7) as a gene therapy for osteoinduction, we investigated in morphological aspect at 1, 2, 4, 6 weeks after cells injection. Primary cultured human dermal fibroblasts, transduced with AdBMP-7, were injected into gastrocnemius muscle of the nude mice. One week after fibroblasts transplantation new tissue was observed in the muscle. Majority of new tissue was evaluated as cartilage and calcification in the matrix was confirmed by Von Kossa stain as well as electron microscopy. Two weeks after transplantation, spongy bone was built up and adipocytes were observed in intertrabecular spaces. Osteoblasts and osteoclasts were observed in the bony tissue surface. In the result of Von Kossa-Van Gieson stain, osteolysis was dominant in bony trabeculae. Bone marrow was established in 4th weeks with intertrabecular space filled up by hematopoietic cells. At the 6th weeks, the number of trabeculae decreased and thickness of the cortical bone was increased. A great part of bone matrix has laminar structure which run paralleled to surface and which included osteocytes and canaliculi. These data demonstrate that cell mediated AdBMP-7 for gene therapy initiate development of cartilage and calcification of matrix within 1 week and complete bone and bone marrow formation within 4 weeks, so then, could be made practical application for promotion of osteoinduction.

Effects of Ovariectomy on Bone Mineral Density and Integrin Expression in Maxilla of Rats / 대한해부학회지

In postmenopausal osteoporosis, estrogen deficiency leads to unbalance of bone metabolism, decreased bone formation and increased bone resorption, and the result is reduced bone mineral density (BMD) and bone stiffness. The processes of bone formation and resorption involves the expression of integrins in anchoragedependent cells, such as osteoblast and osteoclast. The osteoporosis-induced rats frequently demonstrated the loss of trabecular bone volume in the tibia, vertebra and mandible due to estrogen depletion. However, in maxilla, study has been rare because of its anatomical limits. So the objective of this study was to investigate bony change and property of integrin expression in maxilla of osteoporosis-induced rats. 12-week-old female Sprague-Dawley rats were bilaterally ovariectomized (OVX). At 1, 2, 3, 4, 8 and 12 weeks, control and OVX group rats were sacrificed respectively. BMD of maxilla of the rats was measured using dual-energy X-ray absorptiometry (DEXA). And then the histopathologic observation, histomorphometric analysis and immunohistochemistry with CD44, alpha2 integrin, alpha5 integrin, alpha6 integrin, alphav integrin and beta3 integrin were done. BMD of alveolar bone in maxilla was decreased with significance statistically after OVX 4 weeks and was decreased 18.15% at OVX 12 weeks group compared to control group. From OVX 4 to 12 weeks, the thickness of periodontal ligament space was decreased, the number of osteoclast and the size of marrow stroma were increased than control group. By histomorphometric analysis, the size of marrow stroma of alveolar bone in maxilla was increased 86.42% at OVX 12 weeks group compared to control group. CD44 was widely expressed throughout the odontoblast, cementoblast, dental pulp, preiodontal ligament, osteocyte, osteoclast and perivascular tissue at control group, and CD44 immunoreactivity was increased the odontoblast, osteoblast and osteoclast at OVX groups. alpha2 integrin was expressed the odontoblast and osteoblast at control group, but alpha2 integrin immunoreactivity was decreased the osteoblast at OVX 12 weeks group. alpha5 integrin was expressed the cementoblast, osteoblast and osteoclast at control group, and alpha5 integrin immunoreactivity was decreased the osteoblast and was increased the osteoclast from OVX 4 weeks group. alpha6 integrin was weakly expressed the odontoblast, cementoblast, osteoblast and osteoclast at control group, and alpha6 integrin immunoreactivity was weakly increased the osteoclast from OVX 4 weeks. alphav integrin was expressed the odontoblast and osteoclast at control group, and alphav integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. beta3 integrin was expressed the osteocyte and osteoclast at control group, and beta3 integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. From these results, alveolar bone in maxilla of OVX rats was decreased BMD gradually. Moreover, alpha2 and alpha5 integrin expression of osteoblast was decreased, and alpha5, alphav and beta3 integrin expression of osteoclast was increased in OVX rats. Thus, this study indicates that consideration of reduced BMD is necessary in dental procedure of postmenopausal women.

The Effects of Bear Bile on the Expression of the p53 Protein in Cancer Cell Lines / 대한해부학회지

The effects of bear bile on the expression of the p53 protein in cancer cell lines were investigated by Western blot and RT-PCR methods. The p53 protein expression was increased after addition of bear bile in HaCaT, KUMA3, KUMA4 and KUMA6 cell lines that carried mutations n the p53 gene, but it is not ncreased n HCT116 and KUMA5 cell lines with the normal p53 gene as determined by Western blot method. The level of p53 mRNA was not altered in all cell lines after the treatment of bear bile as determined RT-PCR. These results indicate that the increment of the p53 protein after addition of bear bile was caused by increase in the protein stability but not by increase in protein synthesis. It can be concluded that bear bile affects positively only in the cell lines which have genetic variation of the p53 gene, and does not affect in the cell lines which have wild-type p53 gene, on the other hand.

Expressions of BMP Antagonists in Shh -/- ; Gli3 -/- Mutant / 체질인류학회지

Sonic hedgehog (Shh) has been known as an essential morphogen for the generation of motor neuron in developing spinal cord. However, motor neuron can be generated in Shh -/- ; Gli3 -/- or Gli2 -/- ; Gli3 -/- mutants although these mutants don't have Shh signaling. To find out the compensatory mechanism for the generation of motor neuron in Shh -/- ; Gli3 -/- mutant, we studied bone morphogenetic protein (BMP) antagonists including follistatin, flik and noggin, and retinoic acid signaling in this mutant. To study expressions of BMP antagonists, we performed in situ hybridization. To examine an activity of retinoic acid, we measured beta -galactosidase activity in retinoic acid response element (RARE) transgenic mouse. The expression of follistatin was reduced at both levels of forelimb and hindlimb in Shh -/- mutant compared to wild type embryo. It was restored at the level of forelimb but reduced at the level of hindlimb in Shh -/- ; Gli3 -/- mutant compared to wild type. The expression of flik was similar with wild type embryo at both levels of forelimb and hindlimb in Shh -/- mutant. The expression of flik was similar with wild type embryo at the level of forelimb however reduced in hindlimb level in Shh -/- ; Gli3 -/- mutant. The expression of noggin, a BMP antagonist, was increased in Shh -/- mutant. Activity of retinoic acid signaling was not affected in Shh -/- or Shh -/- ; Gli3 -/- mutants. From these results, we conclude that retinoic acid but not follistatin and flik, may be involved in the generation of motor neuron in Shh -/- ; Gli3 -/- mutant.

Distribution Patterns of Involucrin in the Stratum Corneum of the Normal and Psoriatic Artificial Skins / 대한해부학회지

Cornified envelope is highly insoluble structure formed beneath the plasma membrane during terminal differentiation of keratinocytes and is stabilized by cross linking of various proteins, including involucrin, loricrin, and cornifin. Psoriasis is a chronic skin disease characterizing inflammatory reaction and hyperproliferation of keratinocyte. There are some differences in involucrin immunolabelling in stratum corneum between normal and psoriasis epidermis. Labelling was convergent to cornified envelope in psoriasis skin but throughout cytoplasm in normal skin. To compare terminal differentiation patterns of normal and psoriasis keratinocytes, we reconstructed normal and psoriatic artificial skin by using primary cultured keratinocytes from normal and psoriasis skin and then performed immunogold labelling for involucrin in stratum corneum. Psoriatic artificial skin had thin and poorly organized corneal layer. Immunogold labelling for involucrin revealed same pattern of that in vivo by showing throughout cytoplasm in lower layer but convergent cornified envelope in upper layer. Compared with psoriatic artificial skin, normal artificial skin had well organized and thick stratum corneum. Involucrin labelling was throughout cytoplasm in most of corneal layer but convergent to cornified envelope in some uppermost cells. Even though some cells show convergent pattern in normal artificial skin, absolute number of this pattern was no lesser than in artificial psoriatic skin because of normal artificial skin had thick stratum corneum. This result showed there was no difference in involucrin distribution in terminal differentiation of normal and psoriasis keratinocytes in organotypic culture model. It is concluded that although well organized multiple corneal layers are formed in normal artificial skin, they can not reach to full maturation of cornified envelope, and difference of involucrin localization in cornified envelope of psoriasis epidermis is related with not peculiarities of the cells but rapid growing in vivo.

The Property of p53 Gene in Cell Lines of Squamous Cell Carcinoma / 대한해부학회지

The mutation of p53 tumor suppressor gene is the most common genetic variation of primary malignant tumors. The occurrence, progression and reaction for medical management of cancers can be different according to the characteristics of p53 gene, even if they are same kinds of malignant tumors. In this study, the property of p53 gene in 4 kinds of squamous cell carcinoma cell lines were investigated by using immunocytochemistry, PCR-SSCP, sequencing and Western blot methods. As a result, p53 mutation detected in 3 kinds of squamous cell carcinoma cell lines. Namely, it is found that T in codon 176 changed to A, and G in codon 281 changed to A in KUMA3 cell lines; CC in codon 241 changed to TT in KUMA4 cell lines; G in codon 266 changed to T in KUMA6 cell lines. In single nucleotide polymorphism of codon 72 of p53 gene, the genetic variations are Arg/Pro heterozygote in KUMA3 and KUMA4 cell lines; Arg/Arg homozygote in KUMA5 cell lines; Pro/Pro homozygote in KUMA6 cell lines. These results will provide useful data for p53 gene researches of various squamous cell carcinomas.

Characteristics of Mitochondrial Events in Synthetic Bile Acids-induced Apoptosis of Human Osteosarcoma Cells / 체질인류학회지

ABSTRACT: Apoptosis of osteosarcoma cells induced by bile duct derivates, HS-1200 was investigated with relation to mitochodria. HS-1200 induced cytochrome c and Smac/DIABLO release from mitochondria which are major factors related to apoptosis. In these apoptosis processes, release of cytochrome c was not blocked by caspase inhibitor, but release of Smac/DIABLO was blocked. BKA, a kind of PTP (permeablity transition pore) inhibitor, did not block both of them. Interestingly, the alteration of MMP was not observed by means of using JC-1 dye. Although MitoTracker, DiOC-6 and Rhodamine123 were used to confirm previous results, the decrease of MMP was not observed. In order to investigate whether this phenomenon is apoptosis-specific or cell-specific process, genistein was added to cells which usually decreased MMP. After adding genistein, MMP was not decreased, suggesting this phenomenon is cell-specific process. Conclusionally, HS-1200 induced apoptosis of osteosarcoma cells via mitochondria, cytochrome c and Smac/DIABLO were released from mitochondria without decrease of MMP. The release of Smac/DIABLO was dependent of caspase.

Synthetic Bile Acid Derivative HS-1200-induced Apoptosis of Human Osteosarcoma Cells / 대한해부학회지

Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and had anticancer effects. However, it wasn`t discovered those materials have apoptosis induced effects on osteosarcoma cells. The present study was done to examine the synthetic bile acid derivatives induced apoptosis on osteosarcoma cells and such these apoptosis events. The synthetic bile acid derivatives, chenodeoxycholic acid (CDCA) induced the cell death on human osteosarcoma (HOS) cells contrary to ursodeoxycholic acid (UDCA). HS-1200, a synthetic derivative of CDCAs, was chosen to experiment apoptosis events in HOS cells. HOS cells treated with HS-1200 showed nucleus condensation, cytochrom c release, Bax/Bcl-xL alteration, activation of caspase-3 and caspase-activated deoxyribonuclease (CAD), and degradation of poly (ADP-ribose) polymerase (PARP). Though this study needs more investigations, these in vitro data suggest that treatment of the synthetic bile acid derivatives can give medical therapy on HOS cells.

Differential Responses to EGF between the KUMA3 Parental and Variant Cell Lines / 대한해부학회지

EGF has been a representive growth factor for understanding the signal transduction that underlies the biological response of a number of growth factors. EGF plays an important role in the regulation of growth inhibition in the squamous cell carcinoma with over expressed EGF receptors, but the mechanism that determine sensitivity to the growth inhibition by EGF are not well understood. The KUMA3 parental cell line, derived from a squamous cell carcinoma of the human lower lip, was established and variant cell line derived from the parental cell line was acquired by treatment with high dose EGF (200 ng/mL) for 6 months. The KUMA3 parental cells treated with EGF showed marked scattering cells with prominent dendritic processes. On the other hand, EGF treated cells in the variant cell line showed a dense monolayer of ovoid cells similar to untreated cells. Therefore, it was conformed that the variant cell line was resistant to the growth inhibitory effect of EGF. To gain insight into the action mechanism of EGF in the KUMA3 parental and variant cell lines, Western blot analysis was examined to identify transregulation of EGF receptor. In the all parental cell lines treated with EGF, EGF receptor proteins were completely disappeared by lysosomal degradation. However, EGF receptor proteins were shown the periodic appearance according to the time course in the variant cell lines treated with EGF. In Western blot and immunocytochemical analyses, annexin II protein evenly distributed in the cytoplasm relocated to the plasma membrane at 9 hr exposure to EGF and was significantly decreased at 12 hr and 24 hr exposures to EGF in the variant cell line. However, annexin II protein relocated to the nuclear membrane at 9 hr exposure to EGF and was not any change in amount at 12 hr and 24 exposure to EGF in the parental cell line. In conclusion, this study suggests that the retention of colony formation pattern and cell morphology in the variant cell line treated with EGF might be owing to the heterologous desensitization of EGF receptor and exocytosis of annexin II.

Morphological Changes in the Striatum after Dopaminergic Cell Transplantation to Parkinson's Disease Rat Model / 대한해부학회지

Parkinson's disease animal model was developed by the destruction of the striatonigral dopaminergic system. The morphological changes in the dopamine depleted striatum after the transplantation of the fetal mesencephalic dopaminergic neurons or tyrosine hydroxylase cDNA transfected human neural stem cells (C4-TH cells) were studied. Male Sprague-Dawley rats, weighting 250~300 gm, were used. To make unilateral lesion of nigrostriatal tract, 6-OHDA (6 microgram/microliter) was injected into the medial forebrain bundle. Two weeks after the lesion surgery, the effect of the 6-OHDA lesion was assessed by monitoring apomorphine (0.5 mg/kg, s.c)-induced turning behavior and confirmed by the lack of TH-immunoreactivity on tissue sections. Either cell suspension from ventral mesencephalic tissue obtained from embryonic day 14 fetus or C4-TH cells was grafted into the rostral striatum. After grafting, rats were tested with apomorphine every 2 weeks for 6 weeks. The grafted rats showing behavioral recovery were sacrificed and analysed by TH, neuropeptide Y (NPY), and parvalbumin (PV) immuno- histochemistry. TH-immunoreactive (ir) neurons were located around the graft and their processes extended into the striatum. The TH-ir axon terminals made a symmetrical synapse with the dendrites of the striatal neuron. Cell bodies either NPY- or PV-ir striatal neuron were observed around the graft and extended their processes into the graft. TH-ir C4-TH cells were also distributed along the needle track such as the transplanted fetal dopaminergic neurons, but had smaller soma and fewer processes than those. It is concluded that the grafted dopaminergic cells are survived in the dopamine depleted striatum and recovered the rotational behavior of Parkinson's disease animal model.