• 대한수의학회지
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• 제39권3호
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• pp.593-601
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• 1999

This study was designed to evaluate the effect of cyclophosphamide(CY) on diethylnitrosamine(DEN)-induced hepatic tumors in rats. Thirty five male or female Sprague Dawley rats were continiously given water containing 0.01% DEN for 10 weeks and then were given with CY 25mg/rat/day in water for 3, 5, 7 or 9 days. The livers of rats were removed and fixed in 10% buffered neutral formalin. he appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody, p53 antibody and apoptotic kit were investigated. The livers of rats given with CY were grossly brilliant, red-brown color, flexible, and thin border, and stainability of the liver cells were restored microscopically, and the vaccuolated and degenerated regions were differentiated from restored regions. These restored findings also were advanced in control group because of no DEN treatment but tended to be less avanced. In immunohistochemistry, positive cells to PCNA antibody appeared more numerous in control groups than that of CY treated groups. Appearance of positive cells in CY-treated group for 7 days and for 9 days were more numerous than those of CY-treated groups for 3 days and for 5 days, respectively. So these findings suggested that CY suppressed cell proliferations and effects of these action were decreased with CY-treated days. The numbers of positive cells to PCNA antibody were more prominent in hepatocellular carcinoma regions and cholangiocarcinoma regions, and then were ranked as order of large liver cell regions and normal liver cell regions. Also the numbers of the positive cells by apoptotic kit tended to be higher in hepatocellular carcinoma regions and cholangiocarcinoma regions but not uniformly in order in all regions and were much less numbers than those of PCNA positive cells. So immunohistochemical methods using PCNA antibody together than using apoptotic kit alone when anti-carcinogen experiments. Rats with positive cells by p53 antibody were 11 of 15 rats(73.4%) in control groups and 12 of 18 rats(66.7%) in CY treated group, respectively. These positive cells appeared focally in early vacuole-occurring regions and were very low in numbers.

• 대한수의학회지
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• 제21권2호
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• pp.73-79
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• 1981

In order to know the morphology and the function of the fat-storing cells (FSC) of the squirrels which were captured at the Pal-Gong mountain near Taegu in December, 1980 (Group A) and May, 1981 (Group B), respectively, the light and electron microscopic observations were conducted on the liver of the squirrels. The results obtained are as follows: 1. Light microscopically, the size of the lipid droplets in the FSC of group A was uniformly larger than those in the cells of group B, and number of the droplets in the FSC of group A was less than those in the cells of group B, The distribution of the FSC of group A was mainly perilobular area while those of group B was centrolobular and midzonal areas. In this point of view, the FSC of the squirrels was similar to those cells of the hamsters. 2. Electron microscopically, general morphology of the sqirrel's FSC was accorded with those of the other mammals. However, the rouph surfaced endoplasmic reticulum in the FSC of group B was more dilated than those in the cells of group A, and more lipid droplets and pinocytopic vesicles were observed in the FSC of group B than those in the cells of group A. 3. From the above evidences, it could be suggested that the metabolic rates in the FSC of the squirrels collected in the spring were higher than those in the cells of the animals collected in the winter.

• 대한수의학회지
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• 제39권6호
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• pp.1057-1065
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• 1999

The study was designated to investigate the electron microscopic findings following diethylnitrosamine (DEN) treatment in rats. Forty four male (Srague Dawley) rats were continuously given water containing 0.01% DEN for 13 weeks and livers of five rats with more tumor lesions at 16 and 17 weeks after initial treatment were used as EM materials. In transmission electron microscopic findings, most small-sized hepatocytes were active cells containing large mount of organelles, but light (pale staining) hepatocytes among small-sized hepatocytes were injured cells containg disorganized organelles. Tumor cells among small-sized hepatocytes were irregularly arranged and have pleomorphic nuclei containing electron dense chromatin but the organelles in cytoplasm were swelled. Large-sized hepatocytes were active cells with condensed chromatin but the cytoplasm of these cells were pale due to be injured and dilated organelles. Dark hepatocytes were apoptotic cells with homogenous pyknotic nuclei and cytoplasm, and the cytoplasm of these cells contained dilated smooth endoplasmic reticulum (sER) but these sER were non-vesiculated. Cholangiocarninoma cells were crowded and were pale by far less number of organelles in cytoplasm and nuclei. In scanning electron microscopic findings, the lumens of portal veins, bile canaliculi, bile ductules, bile ducts and sinusoids were dilated and have irregular folded inner surface by protruded parenchyma.

• 대한한의학회지
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• 제20권3호
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• pp.105-114
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• 1999

To investigate the anti-complementary and cytotoxic effects of oriental prescription, Kuseonwangdogo, on the proliferation of preadipocyte 3T3- L1 cells, we examined biological effects of Kuseonwangdogo. The results obtained were as follows. 1. After 14 days, the body weight of rats treated with Kuseonwangdogo decreased more than that in the control group (p<0.05). However, the weights of liver, spleen and kidney were unchanged. In serum biochemical test, we examined the level of glucose (GLU) and glutamic pyruvic transaminase (GPT). The levels of GOT and CHOL in serum were decreased remarkably by the administration of Kuseonwangdogo (p<0.05). The haematological examination of the tested group showed significant increment of white blood cells (WBC), hemoglobin concentration (HGB), mean corpuscular hemoglobin (MCH) and monocyte (MO). 2. The effect of Kuseonwangdogo on the proliferation of 3T3-L1 cells was tested by the sulforhodamin B(SRB) assay. The high concentration ($100{\mu}l\;and\;200{\mu}l$) of extracts inhibited the proliferation of 3T3- L1 cells. The p-value was <0.01, respectively. 3. The extract of Kuseonwangdogo showed a potent anti -complementary activity. It was suggested that the active principle may be a kind of polysaccharide molecule. 4. The cytotoxic effects of Kuseonwang dogo and its composing herbs in human liver cells (WRL68) and monkey kidney cells (Vero) were examined by the SRB and 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H- tetrazolium bromide (MTT) assay. Cytotoxic effects were not observed.

• 대한한방내과학회지
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• 제33권4호
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• pp.533-542
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• 2012

Objectives : We try to compared the efficacy of six herbal medicines, Rhizoma Alismatis (RA), Fructus Crataegi (FC), Fructus Lycii (FL), Radix Curcumae (RC), Radix Salviae Miltiorrhizae (RSM), and Herba Artemisiae Scopariae (HAS), constituting KHchunggan-tang which was previously proven to be hepatoprotective on non-alcoholic fatty liver disease with combined properties of cellular steatosis, ROS production, and cytoprotection. Methods : HepG2 cells were pretreated with aqueous extracts of the six herb medicines at concentrations of 1, 10, 50 and 100 ${\mu}g/ml$ each, and treated with 0.5 mM palmitate consecutively. After 21 hrs, cell viability was assessed using MTT assay, and the percentage of cells with sub-G1 DNA content was measured using fluorescence-activated cell sorting after propidium iodide staining. Results : The first three extracts, RA, FC, and FL restored cell viability reduced by palmitate in MTT assay, and RA, FC, FL and RC inhibited palmitate-induced apoptosis in sub-G1 analysis. FL showed relatively weak potential only at tested maximal dose, and RA showed the greatest higher efficacy on this experimental cellular model of nonalcoholic fatty liver disease. Conclusions : According to this comparative experiment, Rhizoma Alismatis seems to have the most powerful potential among the six herbs constituting KHchunggan-tang, and consecutive further study seems to be required for more standardized and effective clinical application of KHchunggan-tang for treatment of non-alcoholic fatty liver disease.

• 한방재활의학과학회지
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• 제28권3호
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• pp.1-11
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• 2018

Objectives This study was designed to evaluate the efficacy of Seungyangjeseup-tang on obesity by using 3T3-L1 cells and high fat diet mice. Methods In vitro, Seungyangjeseup-tang extract (SYJST) (10, 50, 100, 200, 400, $800{\mu}g/mL$) ware added in 3T3-L1 cells. SYJST cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assasy. Adipocyte differentiation was measured by Oil Red-O staining. In vivo, the experimental animals were divided into five groups: normal diet-fed normal group (N), high-fat diet (HFD)-fed control group (Con), HFD+SYJST 100 mg/kg group (SY100), HFD+SYJST 200 mg/kg group (SY200), and HFD+olistat 60 mg/kg as a positive drug control group (Orli). Markers of obesity, such as body weight, liver weight, food intake, serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), liver tissue TC, TG and fecal TC, TG were measured. Results In vitro, cytotoxicity was not significant compared with the control group. 3T3-L1 cell's differentiation was significantly decreased in Oil Red-O staining. In vivo, compared with controls, mice treated with SYJST demonstrate lower body and liver weight, and reduced food intake. In addition, SYJST increased TC, TG in the serum but not significance. And SYJST showed decreasing tendency TC, TG in the liver tissue. Furthermore, SYJST increased TC, TG in the facal but not significance. Conclusions Based on the results above, Seungyangjeseup-tang may reduce adipocyte differentiation, body fat, food intake, liver weight in obesity. This suggests that Seungyangjeseup-tang may be clinically useful in obesity treatment.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2655-2661
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• 2014

Background: Talin-1 is a cytoskeleton protein that participates in cell migration and plays a role in tumor formation, migration, and metastasis in different types of cancer. Chinese investigators have observed that the levels of Talin-1 protein and mRNA expression in HCC tissues are significantly lower than in the adjacent non-cancerous tissue. However, Japanese investigators have reported that Talin-1 is upregulated in HCC. Tln2 as homologous gene of Tln-1, which encodes a very similar protein, but the role of Talin-2 is very little known in primary liver cancer (PLC). We investigated whether the expression of Talin-1 in PLC may be associated with the histological subtype as well as the role of Talin-1 in tumor cell invasion and migration using human hepatocellular carcinoma cell lines. Materials and Methods: We measured the mRNA expression levels of Talin-1 and Talin-2 in five human liver cancer cell lines and normal human liver cell ($LO_2$ cell line) by real-time PCR and the protein expression levels of Talin-1 by Western blot. Migration and invasion of the cells were assessed using transwell assays and cell scratch experiments, respectively, and proliferation was assessed by soft AGAR colony formation. Results: Talin-1 and Talin-2 expression differed significantly between the five human liver cancer cell lines and $LO_2$ cell line (p<0.05). Compared with the $LO_2$ cell line, the invasion and migration capabilities of the five cancer cell lines differed significantly (p<0.05). Similarly, the colony-forming ability differed (p<0.05). Conclusions: High levels of Talin-1 expression are correlated with reduced invasion and migration as well as decreased malignancy in human liver cancer cell lines; the suppression of Talin-1 promotes invasion and migration. In addition, Talin-2 may be correlated with invasion and migration in human hepatocellular carcinoma.

• Advances in Traditional Medicine
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• 제9권1호
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• pp.67-73
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• 2009

Temporary clamping of the portal triad is a common strategy to minimize bleeding during liver transplantation. Increasing evidences suggests that oxygen derived free radicals and reintroduction of oxygen in ischemic tissue lead to ischemic and reperfusion injury (I/R) and lead to apoptosis and necrosis. Adult Wistar rat subjected to 60 min of partial liver ischemia followed by three hour reperfusion. Eighteen Wister rats were divided into sham-operated control group (I) (n = 6), ischemia and reperfusion group (II) (n = 6), folic acid treated group (1 mg/kg body weight/daily by oral route for 7 days before induced ischemia reperfusion maneuver) (III) (n = 6). Apoptotic and necrotic hepatocytes, mitochondrial antioxidant enzymes were measured. Liver injury was assessed by alanine transaminases (ALT), aspartate transaminases (AST), liver histopathology and electron microscopy. An ischemic and reperfusion hepatocellular injury was indicated by increased serum-ALT, AST, histopathology and electron microscopy studies. Apoptotic and necrotic cells were increased which was revealed by flow cytometry in I/R group. Pre- treatment with folic acid significantly decreased serum -ALT, AST levels, apoptotic and necrotic cells after 1 h ischemia followed by 3 h of reperfusion. Histopathology and TEM studies showed markedly diminished hepatocellular injury in folic acid pretreated rats during the hepatic I/R, which reached a level comparable to saline-treated rat of sham operated group. On the basis of our findings it may be concluded that folic acid afforded significant protection from necrosis and apoptosis in I/R injury.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.759-764
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• 2013

Background: The high recurrence rate after hepatic resection in hepatocellular carcinoma (HCC) is a major obstacle to improving prognosis. The objective of the present study was to explore the function of genistein, a soy-derived isoflavone, in enhancing the inhibitory effect of cisplatin on HCC cell proliferation and on tumor recurrence and metastasis in nude mice after curative hepatectomy. Methods: Proliferation of human HCC cells (HCCLM3) was detected by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. Synergistic effects of genistein and cisplatin were evaluated with the median-effect formula. Nude mice bearing human HCC xenografts underwent tumour resection (hepatectomy) 10 days post implantation, then received intraperitoneal administration of genistein or cisplatin alone or the combination of the two drugs. 33 days after surgery, recurrent tumours and pulmonary metastasis were evaluated individually. MMP-2 level in recurrent tumours was detected by immunohistochemistry and real-time PCR; MMP-2 expression in HCCLM3 was detected by immunocytochemistry. Results: Genistein and cisplatin both suppressed the growth and proliferation of HCCLM3 cells. The two drugs exhibited synergistic effects even at relatively low concentrations. In vivo, mice in the combined genistein and cisplatin group had a smaller volume of liver recurrent tumors and fewer pulmonary metastatic foci compared with single drug treated groups. Cisplatin upregulated the expression of MMP-2 in both recurrent tumours and HCCLM3, while genistein abolished cisplatin-induced MMP-2 expression. Conclusions: Genistein reinforced the inhibitory effect of cisplatin on HCC cell proliferation and tumour recurrence and metastasis after curative hepatectomy in nude mice, possibly through mitigation of cisplatin-induced MMP-2 upregulation.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.511-518
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• 2011

To avoid hyperacute rejection of xenografts, ${\alpha}1,3$-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of ${\alpha}1,3$-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that ${\alpha}2,3$-sialyltransferase (${\alpha}2,3ST$) and ${\alpha}2,6$-sialyltransferase (${\alpha}2,6ST$) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of ${\alpha}$-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of ${\alpha}2,6$-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.