• Applied Microscopy
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• v.39 no.2
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• pp.117-124
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• 2009

Clonorchis sinensis is the most important widely distributed parasite of the human bile duct in East Asia and the most prevalent parasitic helminth in Korea. The prevalence rate of human clonorchiasis has remained at about 2.9% in Korea. C. sinensis induces dilatation of the duct, hyperplasia of the mucosa, metaplasia or neoplasia of the mucosal epithelium, periductal inflammation and fibrosis, and thickening of the ductal wall. Fibroblast are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may also contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. In this study, ultrastructural changes, the distribution of lectin receptors and actin protein in cultured SD rat bile duct fibroblast after infection of C. sinensis were observed. Experimental group had been divided into four groups: normal bile duct fibroblast cultured in basal media (G1); C. sinensis infected bile duct fibroblast cultured in basal media (G2); normal bile duct fibroblast cultured in basal media containing excretory-secretory product (ESP) (G1-1); C. sinensis infected bile duct fibroblast cultured in basal media containing ESP (G2-1). Overall, once a host is infected by C. sinensis, it affects the host to the extent that sialic acid of ductal fibroblast is increased. Number of cytoplasmic process of SD rat bile duct fibroblast was increased. Actin protein and sialic acid were located in cell surface. Fibroblast induced by C. sinensis was not recovered to normal fibroblast. The cytoplasm bulk and cytoplasmic process were increased whereas the growth rate of the fibroblast of infected SD rat was reduced rather than that of normal fibroblast. In result, it inhibits fibroblast proliferation and increases actin protein on fibroblast cytoplasm, and so causes fibroblast metamorphosis and cellular mutation.

• Journal of Acupuncture Research
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• v.19 no.2
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• pp.51-64
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• 2002

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.

• Applied Microscopy
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• v.24 no.1
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• pp.28-40
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• 1994

Xerostomia and xerophthalmia are delicate or serous side effects, occuring when the radiotherapy is administered to the head and neck cancer patient. It is known that the cause of the above side effect is radiosensitivity of serous cells. In this study, the ultrastructural features of the parotid glands of the X-irradiated rats were observed. Sprague-Dawley rats weighing 200-250g each were anesthetized with sodium thiopental, and placed on the Mitsubishi linear accelerator. Only the head and neck areas of animals were exposured at the distance of 80cm, within the area of $30X30cm$, in the depth of 1cm, with the speed of 200R/min. Total doses applied were 3,000R or 6,000R depending on the experimental groups. Animals were sacrificed on the 6th hour, 2nd day and 6th day after the irradiation. Parotid glands were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, and followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture, and ultrathin sections were cut. Sections were contrasted with the solution of uranyl acetate and lead citrate, and observed with JEM 100 CX-II electron microscope. The results were as follows: 1. Normal parotid acinar cells are two types; the light and the dark acinar cells. The light acinar cell contains dense secretory granules, whereas dark acinar cells contains granules of medium density with some darker spots within them, or other cells contain granules of medium density with darker rims. 2. Six hours after the irradiation, many acinar cells were degenerated showing variable stages of cytolytic bodies, light bodies, or dense degenerations. Within the acinar cell, Golgi apparatus and granular endoplasmic reticula were most severely altered elements. Granules showed more contrasting densities and irregularities. 3. Two days after the irradiation, some cytolytic bodies, and focal lucent degeneration of cytoplasm, and fine granular alteration of cytoplasmic matrix were pronounced. But other elements including secretory granules are rather looked unlatered. 4. Six days after the irradiation, most severe alterations were seen. Many intracellular canaliculi (or secretion figures), quanta of cytoplasm containing secretion antecedants, severely irregular luminal border, and again contrasting density of secretory granules showing tigroid spots or dense rims were noted. And myoepithelial degenerations were observed not uncommonly. 5. Irregular densities of secretory granules were interpreted as abnormal components of protein or carbohydrate portion are synthesized or abnormally metabolized under severe X-irradiation. 6. Myoepithelial degeneration and related alteration of nerve endings, etc., were suggested as the other causes of xerostomia following X-irradiation. 7. It is requested that radiation doses should be arranged, considering in mind not only the sensitivity of acinar cells but also the myoepithelial and neural functions.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.177-181
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• 1994

The sequences of the cytoplasmic 5S rRNAs(EMBL accession number X73589 and X73890) from two polupores, Ganoderma applanatum and Schizopora paradoxa, were determined by the direct chemical method for sequencing RNA and compared to the sequences of 9 reported mushrooms. 5S rRNAs of Ganoderma applanatum and Schizopora paradoxa consisted of 118 bases and fit the secondary structure model of the 5S rRNAs of basidiomycetes proposed by Huysmans et al. Based on Kimura’s K_nuc values, the closest fungus to Ganoderma applanatum was Ceratobasidium cornigerum and the one to Schizopora paradoxa was Bjerkandera adusta. When the secondary structures of 5S rRNAs of 11 mushrooms were compared the base substitution occurred at helix regions more than at loop regions. When a phylogenetic tree was constructed using the Neighbor program of the PHYLIP package, it partially discriminated and separated the mushrooms of the Hymenomycetes by the order.

• Applied Microscopy
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• v.23 no.1
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• pp.35-55
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• 1993

cis-Dichlorodiammineplatinum (II) (cis-Platin), a metallic compound, has widely been used as an effective anticancer chemotherapeutic agent. The precise mechanism of action of this agent is still unknown, but it is postulated that cis-Platin may act on the cancer cell like bifunctional alkylating agents. Although this agent is very beneficial to the patients with cervical cancer, germinoma of testis, neuroblastoma and others, it may also damage to the normal cell so that many side effects; severe hemorrhagic enterocolitis, bone marrow depression, renal damage and liver damage will develope. This experiment has been undertaken to pursue the cytotoxic effects of the cis-Platin on the ultrastructures of the interalveolar septum in the mouse lung. A total of 55 healthy male mice of ICR strain were used as experimental animals and divided into 5 mice of normal control group and 50 mice of cis-Platin treated group. The mice of cis-Platin treated group were sacrificed by carotid exsanguination at 6, 12, 24 hours, 3 days and 7 days after intraperitoneal injection of 6.0 mg of cis-Platin ($Abiplatin^R$ Abic Co. Ltd.) per kg of mouse body weight. The specimen obtained from the lower lobe of left lung were sliced into $1mm^3$ and prefixed with 2% glutaraldehyde -2.5% paraformaldehyde solution prepared with Millonig's phosphatae buffer solution (pH 7.4) at $4^{\circ}C$ for 3-4 hours. After postfixation with 1% osmium tetroxide solution all specimens were embedded in Epon 812. Ultrathin sections about $600-800{\AA}$ in thickness were stained with uranyl acetate and lead citrate and observed with Hitachi-600 electron microscope. The results obtained were as follows: 1. Local swellings with increase of electron density and number of pinocytic vesicles in the cytoplasms of the type I pneumocyte and endothelial cell of the blood air barrier in interalveolar septum of cis-platin treated mice were observed. 2. Cisternae of rough endoplasmic reticulum were dilated and sacculated in association with detachment of membrane bound ribosomes of the type II pneumocyte in interalveolar septum of cis-Platin treated mice. 3. Swollon mitochondria with uneven electron density of their matrix were observed in the type II pneumocyte of interalveolar septum in the cis-Platin treated mice. 4. The lamellae of lammelar bodies in type II pneumocyte of interalveolar septum in cis-Platin treated mice were devoided or transformed into homogeneous electron dense material. It is consequently suggested that cis-Platin would induce the cellular edema of type I pneumocyte and endothelial cell, and degenerative changes of cytoplasmic organelles of the type II pneumocyte in the interalveolar septum of the mouse lung.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.221-232
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• 1993

Morphological changes in the uterine and vaginal epithelial cells of the Korean native goats were studied in fifteen primiparous goats slaughtered on the day of parturition and on days 1, 3, 10 and 21 postpartum. 15 uterus and vagina from goats were examined by scanning and transmission electron microscopy. The results obtained in this study were summarized as follows : 1. Transmission electron microscopically, long microvilli which sometimes ramified were found until 10 days postpartum, while short microvilli were found at 21 days. The high electron dense irregular-shaped mitochondria were found in the cytoplasm and the crystalline structure of the mitochondrial matrix was also found from 1 day to 10 days postpartum. Well-developed rough-endoplasmic reticulum (rER) with dilated cisternae which contained the proteins materials was observed at 21 days postpartum. These materials were fused each other and then large granules were found in the free surface of the cytoplasm. A few lipid droplets were generally appeared in the cytoplasm, while numerous droplets were found at 21 days postpartum. A moderate number of ribosomes, a few multivesicular bodies, vesicles, lysosomes and macrophages were found. The globule leucocytes were observed from 0 to 3 days postpartum by transmission electron microscopy. The short microvilli, high electron dense cytoplasm and severe indentation of the nuclear enbelope were found in the vaginal epithelium. Numerouos small vesicles and a few vacuoles were observed in the apical cytoplasmic portion of the epithelium. A few mitochondria were high electron dense and irregular in shape. A moderate amounts of microfilaments, loose intercellular space and dilated rER were also found at 21 days postpartum. 2. Scanning electron microscopically, the folds of the uterine mucosa were generally deep. The long microvilli of the epithelium were found until 3 days postpartum, while short microvili were found at 10 and 21 days postpartum. The distinct intercellular boundary was seen. The apporcine secretory profile of the epithelium observed at between 3 and 10 days postpartum and the cells were somewhat protruded into the lumen. The short microvilli were found on the surface of the protruded cells, while polygonal microridge profile of the epithelium and some dome-shaped epithelium were also observed at 21 days postpartum. The folds of the vaginal mucosa were deep and epithelium was polygonal in shape. The microvilli of the epithelium were long until 3 days postpartum, while they were short at 10 and 21 days. The polygonal epithelium was invaginated into the center of the cell surface until 10 days postpartum. The microridge and dome in shape of the epithelium were found at 10 days postpartum, while the polygonal and exfoliating epithelium were observed at 21 days.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.403-407
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• 2000

Ultrastructure and function of testis somatic cells in freshwater prawns Macrobrachium nipponense were studied. The paired testes of the prawn were elongated, united at their anterior end, which lay between the dorsal surface of the hepatopancreas and the heart. Each testis consisted of a large number of seminiferous cords compactly held together by connective tissue. A seminiferous cord was composed of an outer layer of simple squamous epithelium, a basement membrane, the closely packed germ cells and sustentacular cells of the germinal ridge, and an inner layer of simple cuboidal epithelial cells. Leydig cell-like cells in an angular areas filling the space of the seminiferous cords were observed. The nuclei of leydig cell-like cells were characterized by a distinct nucleolus. The simple squamous epithelial layer was composed of flattened cells tying on a basement membrane. The nuclei of the flattened cells were often overlapped in a layer, and the cytoplasm of the cells was observed just near the nuclei. The sustentacular cells were complex in morphology. These cells had relatively small cell bodies from which long cytoplasmic extensions ramified reached the space of germ cells in the germinal ridge. The nuclei of sustentacular cells usually exhibited angular profiles and located most commonly at the periphery of the cords. Cells of simple cuboidal epithelium located between germinal ridge and lumen of seminiferous cord, and part of the cells were adjacent to basal lamina, The cuboidal epithelial cells contained numerous mitochondria, the well-developed rER, the well-developed Golgi complex, and irregularly shaped nuclei. Transition vesicles appeared on the cis side of the Golgi complex. The large vesicles on the trans side of the complex appeared to fuse to form a membrane-bound structure. A number of pits on the cell apex suggested exocytotic activity for secretion of the sperm supporting matrix.

• Nutritional Sciences
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• v.9 no.4
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• pp.267-272
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• 2006

The effect of methanol extract from Agaricus blazei Murill on the hepatotoxicity was investigated $CCl_4$ is one of the oldest and most widely used toxins for the induction of hepatic damages and fibrosis in experimental animals. In this study, male Sprague-Dawley(SD) rats were randomly divided into 6 groups of the control(C), $CCl_4(T),\;CCl_4$ and high fat group(TL) with matching sub-groups of Agaricus blazei Murill extract-fed groups of CA, TA and TLA. Methanol extracts of Agaricus blazei Murill were fed 50 mg/kg B.W daily via drinking water. A 1.2 mL of $CCl_4/kg$ body weight was administered by oral intubation twice a week for total of six times. The levels of total-cholesterol, TG, LDL and LDL-phospholipids were elevated by $CCl_4$ treatment as compared to the control(C). However, Agaricus blazei Murill methanol extract feeding in the group of TA and TLA significantly(p<0.05) decreased TG by 53.1 % and 17.9% compared to the internal control of T and TL, respectively. Triglyceride of TL was increased by 3.33 times(p<0.05) compared to the control(C) with $CCl_4$ and high fat administration from 3.78 mg/g to 12.60 mg/g liver. The extract(CA) also reduced kidney weight compared to the control(C). With the administration of high fat and $CCl_4$(TLA), the extract reduced the organ weight of both liver and kidney and further, significantly reduced TG, total cholesterol and GTP activity. Hepatoprotective effects of Agaricus blazei Murill on GOT, GPT, AP and LDH activities were enhanced by the extract feeding. Electronmicrograph showed that $CCl_4$ deteriorated the structure of cytoplasmic matrix with its uneven distribution. However, the extract reconstituted the damaged cytoplasm and stimulated mitochondriogenesis. The above results suggest that Agaricus blazei Murill may have a possible protective effect against chemically induced liver damage and further help to reduce the symptoms of fatty liver.

• Animal Bioscience
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• v.34 no.6
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• pp.975-984
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• 2021

Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

• Applied Microscopy
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• v.40 no.2
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• pp.89-99
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• 2010

Fibroblasts are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. Mitochondria produce ATP through oxidative metabolism to provide energy to the cell under physiological conditions. Also, mitochondrial dysfunction and oxidative stress have been implicated in cellular senescence and aging. Alternations in mitochondrial structure and function are early events of programmed cell death or apoptosis and mitochondria appear to be a central regulator of apoptosis in most somatic cell. Clonorchis sinensis, one of the most important parasite of the human bile duct in East Asia, arouses epithelial hyperplasia and ductal fibrosis. Isolated fibroblast from the bile ducts of rats infected by C. sinensis showed increase of cytoplasmic process. In addition, decrease of cellular proliferation was observed in fibroblasts which was isolated from normal rat bile duct and then cultured in media containing C. sinensis excretory-secretory product. However, the effects of C. sinensis infection on the mitochondrial enzyme distribution is not clearly reported yet. Therefore, we investigated the structural change of C. sinensis infected bile duct and mitochondrial enzyme distribution of the cultured fibroblast isolated from the C. sinensis infected rat bile duct. As a result, C. sinensis infected SD rat bile ducts showed the features of chronic clonorchiasis, such as ductal connective and epithelial tissue dilatation, or ductal fibrosis. In addition, fibroblast in ductal connective tissue was damaged by physical effect of fibrotic tissue and chemical stimulation. Immunohistochemically detected mitochondrial electron transferase (ATPase, COXII, Porin) was decreased in C. sinensis infected rat bile duct and cultured fibroblast from infected rat bile duct. It can be hypothesized that the reason why number of electron transferase decrease in fibroblast isolated from the rat bile duct infected with C. sinensis is because dysfunction of electron transport system is occurred mitochondrial dysfunction, increase of ROS (reactive oxygen species) and apoptosis after chemical damage on the cell caused by C. sinensis infection. Overall, C. sinensis infection induces fibrotic change of ductal connective tissue, mutation of cellular metabolism in fibroblast and mitochondrial dysfunction. Consequently, ductal fibrosis inhibits fibroblast proliferation and decreases mitochondrial electron transferase on fibroblast cytoplasm. It was assumed that the structure of bile duct could not normalized and ductal fibrosis was maintained for a long period of time according to fibroblast metamorphosis and death induced by mitochondrial dysfunction.