• Applied Microscopy
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• 제26권2호
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• pp.207-219
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• 1996

This study was designed to investigate the ultrastructural alterations of the hepatocyte and bile canaliculus of the fasted mice with transmission and scanning electron microscopes. The morphometry was also carried out for the caliber of the bile canaliculus and the number, length and thickness of the microvillus. The hepatocyte observed in the three day fasting group showed ultrastructural images of active function. The dilated bile canaliculi, especially of type II were increased in number as compared with those seen in the normal group. However, the hepatocyte observed in the six day fasting group showed ultrastructural images of inactive function. The bile canaliculi without dilation (type I) were increased in number. The number of microvilli were identical with one another among the different types of bile canaliculi, while their length and thickness were reduced in the dilated bile canaliculi. From the evidence, the luminal size of the bile canaliculi seems to be easily changeable according to the functional state of the hepatocyte. However, the microvilli may not be changed in number but may be changed length and thickness when the bile canaliculi are dilated.

• Applied Microscopy
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• 제28권1호
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• pp.83-90
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• 1998

본 연구에서는 박절편과 동결할단복제법을 이용하여 흰쥐 간세포에서 dehydrocholic acid가 수송되는 경로를 전자현미경적으로 조사하고자 하였다. 정상군이나 dehydrocholic acid 투여군에서 대부분의 Golgi 장치는 형성면을 담세관으로 향하고 있었다. Dehydrocholic acid 투여 20분 후에 세포질내세망과 Golgi 장치 및 소포 등이 담세관 주위에 증가되어 있었는데 특히 Golgi 장치 형성면에서는 소포가 될 것으로 추정되는 싹이 돌출되어 있었으며 소포들은 담세관에 융합된 것들도 관찰되었다. 이러한 소견으로 미루어 담즙산의 분비는 Golgi 장치 형성면의 쌀이 유리되어 형성된 소포가 담세관막에 융합되므로서 이루어질 것으로 추정된다.

• Applied Microscopy
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• 제22권2호
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• pp.46-65
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• 1992

In this study, we have made morphological and cytochemical observations to investigate the type of Golgi apparatus around the bile canaliculus. The animal (Wister, $220{\sim}250gm$) were divided into 4 groups; normal, hydrochol, colchicine and hydrochol-colchicine. The Golgi apparatus is classified into 16 different types from 4 different groups. In the normal group, we could observe 12 different types of the sixteen. Type I which showed convexed cisterns facing the bile canaliculi was most abundant of the types. In the hydrochol group, 14 types were observed. Type VII and type I showed convexed cisterns facing the bile canaliculus and were abundant. In the colchicine group, 11 different types were viewed and type XIV which showed intensely dilated cisterns without the polarity was predominant. In the hydrochol-colchicine group, we observed 3 different types. Type XIV clearly showed the highest percentage, although that type was less numerous in this group than in the colchicine group. In the hydrochol group, the Golgi apparatus showed a tendency to increase in numbers, while in the hydrochol-colchicine group the Golgi apparatus showed a tendency to decrease in numbers. The reactive products of thiamine pyrophosphatase and acid phosphatase were apparent over the distal Golgi cistern in the normal and hydrochol groups, but were decreased or not observed in the colchicine and hydrochol-colchicine groups. From the results, it is assumed that with the presence of the microtubule, Golgi cisterns are dilated with polarity after stimulation of secretion. Without the microtubule, the cistern becomes more intensely dilated and none polaric. Also the enzymes within the cisternal membrane become decreaed or absent and the Golgi apparatus decreases in numbers after activation of secretion.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제19권1호
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• pp.1-11
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• 2016

Cholestasis results from impairment in the excretion of bile, which may be due to mechanical obstruction of bile flow or impairment of excretion of bile components into the bile canaliculus. When present, cholestasis warrants prompt diagnosis and treatment. The differential diagnosis of cholestasis beyond the neonatal period is broad and includes congenital and acquired etiologies. It is imperative that the clinician differentiates between intrahepatic and extrahepatic origin of cholestasis. Treatment may be supportive or curative and depends on the etiology. Recent literature shows that optimal nutritional and medical support also plays an integral role in the management of pediatric patients with chronic cholestasis. This review will provide a broad overview of the pathophysiology, diagnostic approach, and management of cholestasis beyond the neonatal and infancy periods.

• Applied Microscopy
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• 제23권2호
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• pp.53-77
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• 1993

In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

• 대한약리학회지
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• 제18권1호
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• pp.43-54
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• 1982

Bile formation is a complex process comprised of three separate physiologic mechanism operating at two anatomical sites. At present time, it was known that at least two processes are responsible for total canalicular secretion at the bile canaliculus. One of the processes is bile salt-dependent secretion (BSDS) hypothesis that the active transport of bile salts from plasma to bile provided a primary stimulus for bile formation: the osmotic effect of actively transported bile acid was responsible for the movement of water and ions into bile. The other process is bile salt-independent secretion (ESIS), which is unrelated to bile salt secretion at the canaliculus and which may involve the active transport of sodium. The third process for bile formation involves the biliary ductal epithelium. Secretin-stimulated bile characteristically contained bicarbonate in high concentration. Therefor, it was suggested that secretin stimulated water and bicarbonate secretion from the biliary ductules. One the other hand, it was found that a large amounts of cAMP was present in canine bile but no apparent relationship between bile salt secretion and cAMP content in dog bile. However, bile flow studies in human have demonstrated that secretin and glucagon increase bile cAMP secretion as does secretin in baboons. Secretin increases baboon bile duct mucosal cAMP levels in addition to bile CAMP levels suggesting that in that species secretin-stimulated bile flow may be cAMP mediated. It has been postulated that glucagon and theophylline which increase the bile salt-independent secretion in dogs might act through an increased in liver cAMP content. In a few studies, the possible role of cAMP on bile formation has teen tested by administration of an exogenous derivative of cAMP, dibutyryl cAMP. In the rat, DB cAMP did not modify bile flow, but injection of DB cAMP in the dog promoted an increase in the bile salt-independent secretion. Because of these contradictory results, this study was carried out to examine the relationship between cyclic nucleotides and bile flow due to various bile salts as well as secretin or theophylline. Experiments were performed in rabbits with anesthesia produced by the injection of seconal(30 mg/kg). Rabbits had the cystic duct ligated and the proximal end of the divided common duct cannulated with an appropriately sized polyethylene catheter. A similar catheter was placed into the inferior vena cava for administration of drugs. Bile was collected for determination of cyclic nucleotides and total cholate in 15 min. intervals for a few hours. The results are summerized as followings. 1) Administrations of taurocholic acid or chenodeoxycholic acid increased significantly the concentrations of cAMP and cGMP in bile of rabbits. 2) Concentration of cAMP in bile during the continuous infusion of ursodeoxycholic acid, was remarkedly increased in accordance with the increase of bile flow, while on the contrary concentration of cGMP in bile was decreased significantly. 3) Dehydrocholic acid and deoxycholic acid significantly increased bile flow, total cholate output and cyclic nucleotides in bile. 4) Only cAMP concentration in bile was significantly increased from control value by secretin, while theophylline increased cAMP as well as cGMP in rabbit bile. 5) In addition, the administration of secretin to taurocholic acid-stimulated bile flow increased cAMP while theophylline produced the increases of cAMP and cGMP in bile. 6) The administration of insulin to taurocholic acid-stimulated bile flow decreased cAMP concentration, while on the contrary cGMP was remarkedly increased in rabbit bile.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.695-701
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• 2013

Opisthorchis viverrini infection causes inflammation and liver injury leading to periductal fibrosis. Little is known about the pathological alterations in bile canaliculi in opisthorchiasis. This study aimed to investigate bile canalicular alterations in O. viverrini-infected hamsters and to examine the chemopreventive effects of curcumin on such changes. Hamsters were infected with O. viverrini and one group of animals was fed with 1% dietary curcumin supplement. Animals were examined during the acute infection phase, days 21 and 30 post-infection (PI) and chronic infection phase (day 90 PI). Scanning electron microscopy revealed that in the infected group fed with a normal diet, bile canaliculi became slightly tortuous by 30 day PI and more tortuous at day 90 PI. Transmission electron microscopy showed a reduction in microvilli density of canaliculi starting at day 30 PI, with a marked loss of microvilli at day 90 PI. These ultrastructral changes were slightly seen at day 21 PI, which was similar to that found in infected animals fed with 1% curcumin-supplemented diet. Notably, curcumin treatment prevented the reduction of microvilli density, reduced the dilation of bile canaliculi, and decreased the tortuosity of the bile canaliculi relative to non-infected animals on a normal diet at days 30 and 90 PI. These results suggest that curcumin reduces alteration of bile canaliculi and may be a promising agent to prevent the onset of bile duct abnormalities induced by O. viverrini infection.

• 대한수의학회지
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• 제35권4호
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• pp.659-670
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• 1995

Cyclosporin A extracted from fungus Trichoderma polysporum Rifai and Cyclindrocarpon lucidum Booth serves as an important immunosuppressive drug in transplantation surgery. Systemic treatment with cyclosporin A induces an impairment of the biliary excretion of the bile salts and cholestasis. This study was designed to observe the Ultrastural changes of the hepatocytes and the bile canaliculi in cyclosporin A-induced intrahepatic cholestasis in rats. Cyclosporin A was injected into male Wistar rats intraperitoneally 50mg per kg body weight and rats were necropsied at 1, 3, 6, 9, 12, 24 hours. The liver tissues were observed with transmission and scanning electron microscopes and the results were as follows. Transmission electron microscopy: After cyclosporin A injection, SER and lysosomes were increased in the hepatocytes until 9 hours. At 12 hours after injection of cyclosporin A, RER with dilated cistern were increased, and SER, lysosomes in the cytoplasm were decreased. From 1 hour to 24 hours after injection of cyclosporin A, there were dilation of bile canalliculi and decreased or lost microvilli. At 24 hours the dilation of bile canaliculi were decreased. Scanning electron microsocopy: After cyclosporin A injection, the bile canaliculi were dilated and the microvilli were shortened, decreased or lost according to the sites. At 24 hours, the microvilli packing the bile canaliculi were observed. These observations suggest that cyclosporin A-induced cholestasis is associated with the dilation of bile canaliculi, increased microfilaments of the pericanalicular region and decreased or lost microvilli.

• Applied Microscopy
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• 제27권3호
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• pp.295-308
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• 1997

본 연구에서는 dehydrocholic acid와 biligrafin 투여 20분 후, 기아 8일 후 및 담관결찰 48시간 후의 흰쥐 간을 동결할단하여 담세관 주위 폐쇠띠의 미세구조를 관찰하였다. 정상대조군에서 폐쇠띠는 여러부위에서 $1\sim4$개의 줄로 이루어진 망상구조를 보였다. dehydrocholic acid 투여군에서는 폐쇠띠의 폭이 넓어져서 망상구조의 줄은 $6\sim9$개로 증가된 부위가 많이 나타났다. 그러나 biligrafin 투여군에서는 줄이 감축되어 있었으며 망상구조는 흩으러져 있었다. 기아군에서는 많은 부위에서 폐쇠띠가 $2\sim3$개의 줄로 이루어져 있었으며 이들 줄은 느슨하게 연결되어 있었다. 결찰군에서도 많은 부위에서 폐쇠띠의 망상구조는 흩으러져 있었으며 줄들이 감축되어 있거나 아직 감축되지 않은 상태로 있는 부위도 관찰되었다. 그러나 모든군에서 줄들은 틈을 보이면서 단절되어 나타나거나 선상으로 배열된 입자로 이루어져 있었는데 특히 biligrafin군과 결찰군에서 이러한 소견이 현저하였다. 줄의 끝이 유리되어 있는 것들은 기아군에서 많이 관찰되었는데 dehydrocholic acid군에서는 줄의 끝이 교통반에 접해 있거나 그 주위를 둘러싸고 있는 것들을 많이 볼 수 있었다. 줄들이 불연속적으로 나타나거나 끝이 유리된 것은 정상군에서도 드물지 않게 관찰되었다. 결찰군을 제외한 모든 군에서 담세관 주위 폐쇠띠가 그망안에 입자의 집괴를 함유하고 있었다. 이상의 증거로 미루어 담세관 주위의 폐쇠띠는 담세판의 상태에 따라 쉽게 변하는 것 같이 보이며 같은 담세관에서 일지라도 부위에 따라 구조적 변화에 차이가 있는데 이러한 소견은 정상상태하에서도 비슷하게 나타나고 있어 폐쇠띠가 완전한 장벽으로서의 역할을 하고 있지 않는 경우도 있는 것 같이 보인다. 또한 폐쇠띠는 기계적 자극이나 다른 어떠한 인자에 의해서 쉽게 영향을 받는 것 같이 보이며 폐쇠띠의 변화에 연관하여 줄과 입자의 집괴 사이에도 어떠한 관계가 있을 것으로 추정된다.

• Applied Microscopy
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• 제26권4호
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• pp.401-410
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• 1996

The microwave fixator has recently been introduced in morphological research. The present study was carried out to investigate the ultrastructural effects of microwave fixation of rat brain. kidney, liver and skeletal muscle tissues. The results are as follows: In the case of microwave fixed cerebrum. the cytoplasmic processes of neurons and the various membranous organelles such as nuclear envelope, mitochondria, rough endoplasmic reticulum and Golgi apparatus were well preserved, The myelin sheath wrapping neuronal axon was prominent. Microwave fixed hepatocytes showed the microvilli on the free surface of bile canaliculus, the evident nucleolar components, and typical organelles. In nephron, ultrastructures of glomerulus and Bowman's capsule were preserved, and also tubular wall were structurally observed. Among the skeletal muscle cells, plentiful collagen fibers were appeared, myofibrils and mitochondria were typically observed. In conclusion, the microwave fixation procedures result in an good preservation of the tissues and would be time- and reagent-saving.