• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• Radiation Oncology Journal
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• v.17 no.2
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• pp.151-157
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• 1999

Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

• Journal of Society of Preventive Korean Medicine
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• v.8 no.2
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• pp.141-156
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• 2004

Objectives : JianTeng-Yuan(交藤圓) is said to be a prescription for preservation of health in ${\ulcorner}$HuaTuo ZhongZangJing(華陀 中藏經)${\lrcorner}$. It is known to have the effect of Bu-Shen(補腎: strengthening kidney) and Yi-Shou(益壽: prolonging the span of one's life). This study investigates whether JTY is effective on inhibition of oxidation stress. Methods : Sprague-Dawley Rats(12-week-old, weight $300{\pm}20g$) were divided into 3 groups. Normal group(n=8) was injected PBS(1ml/body, s.c) at the back neck's skin. Control group(n=8) was injected D-galactose(50mg/kg, 1ml PBS/body, s.c) to induce pathological animals. JTY group was injected the same treatment for the Control group, and fed containing JTY(10%). The whole groups were treated 1 time per day for 6 weeks. After rats were sacrificed and anti-oxidant enzyme(SOD, CAT, G-px) activity, GSH quantity of RBC and tissue(heart, liver and kidney), plasma Vit-C quantity were examined. Besides, the MDA levels of liver and kidney, lipofuscin of heart and endurance of erythrocyte membrane were measured. Results : In the JTY group, RBC's SOD activity decline was halted by 21% of the normal level, compared to the control group ; G-px activity(unit/g of Hb) increased significantly, compared to the normal group ; and the level of Vit-C in plasma increased by 16%. Heart's SOD activity was kept at the same level as that of the normal group ; and CAT activity decline was halted by 26%. Kidney's CAT and G-px activities were kept at the same level as that shown in the normal group, implying the existence of halting effect. Liver also showed a slight halting effect against the decline of anti-oxidant ability, but the effect was not significant(a=0.05). A comparison between the levels of peroxide in SD rats showed that the level of TBARS in plasma increased significantly in the control group and that it was normal in the JTY group. The livers in the JTY group, compared to those in the control group, showed 36% halting effect of the normal level while their kidney's indicated the level significantly lower than the normal level. Heart's lipofuscin increased significantly in the control group, but was alike in both the JTY and the normal groups. Endurance of erythrocyte membrane(%) decreased significantly in the control group while it was kept at the similar level in both the JTY and the normal groups, indicating the halting effect. Conclusions : This study suggests that JTY is effective to defend oxidation stress caused by D-galactose in the animals. It showed that the anti-oxidant ability was maintained and strengthened. On the other hand, it reduced the level of peroxide in animals. In sum, JTY appeared to have the equilibrium normal physiological function in SD rat.

• Journal of Chest Surgery
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• v.38 no.7 s.252
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• pp.461-467
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• 2005

Pleura, diaphragm, pericardial fat pad, intercostal muscles and omentum can be used to protect and revascularize the bronchial suture line of tracheal transplantation, lung transplantation and pulmonary resection. The purpose of the present study is to compare the influence of the pleura, diaphragm and omentum in survival of isolated tracheal segments in the experimental animals. Material and Method: Sprague-Dawley rats weighing 250- 350g were used. The animals were divided in three groups; the pleura, omentum and diaphragm. Following intraperitoneal anesthesia, endotracheal intubation was performed. Then the trachea was exposed. A three-ring sec- tion of cervical trachea was excised. The resected trachea was implanted at each sites. After 2 weeks, rats were sacrificed. Histopathological examination of the tracheal segments was performed. For comparison of each groups, histopathological viability of resected tracheal segment was scored by three tissue layers; epithelium, submucosa, and cartilage. The results were presented as average score. Result: In histopathological examination, submucosa and cartilage using tracheal segment necrosis scoring system. The pleural group showed well preserved tissue. There was minimal necrosis and inflammation compared with other groups. In the pleural group, tracheal necrosis scores were $2.17\pm0.983$at epithelium, $1.67\pm0.516$ at submucosa and $2.17\pm0.753$ at cartilage. At the omental group, scores were $1.00\pm0.00,\;1.60\pm0.548\;and\;1.80\m0.447$. In the diaphragmatic group, scores were $1.40:\pm0.894,\;2.40\pm0.547\;and\;2.20\pm0.447$. Total necrosis score were $6.00\pm1.789$ in the pleural group, $4.40\pm0.894$ in the omental group and $6.00\pm1.414$ in the diaphragmatic group. Conclusion: There were no significant viability differences in terms of total necrosis score for the viability of resected tracheal segment. But the best result was achieved in the omental group. Therefore, omental wrapping on tracheal graft site will be beneficial for the prevention of graft necrosis.

• Journal of Life Science
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• v.19 no.1
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• pp.21-33
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• 2009

In this study, we investigated the effect of Rhei Rhizoma (RR; 大黃) water extract on gene expression in a hypoxia model of cultured rat hippocampal neurons. RR water extract $(2.5{\mu}g/ml)$ was added to the culture media on day 10 in vitro (DIV10), and a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 h) was given on DIV13. After maintaining the cultures in normoxia for 24 hr, total RNA was isolated and used for microarray analysis. The MA-plot indicated that most genes were up- or downregulated within 2-fold. There were more downregulated genes (725 ea) than upregulated ones (472 ea) when larger than Global M value 0.2 (i.e., >15% increase) or smaller than Global M value -0.2 (i.e., >15% decrease) were considered. Antiapoptosis genes such as Tegt (2.4-fold), Nfkb1 (2.4-fold) Veg (1.8-fold), Ngfr (1.6-fold) were upregulated, while pro-apoptosis genes such as Bad (-64%), Cstb (-66%) were downregulated. Genes for combating environmental stress (stress response genes) such as Defb3 (2.7-fold), Cygb (2.2-fold), Ahsg (2.18-fold), Alox5 (2-fold) were upregulated. Genes for cell proliferation (cell cycle-related genes) such as Erbb2 (1.84-fold), Mapk12 gene (1.8-fold) was upregulated. Therefore, RR water extracts upregulate many pro-survival genes while downregulating many pro-death genes. It is interpreted that these genes, in combination with other regulated genes, can promote neuronal survival in a stress such as hypoxia.

• Journal of Life Science
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• v.24 no.4
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• pp.343-351
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• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

• Journal of Food Hygiene and Safety
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• v.25 no.3
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• pp.263-268
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• 2010

This study was demonstrate a repeated oral dose toxicity of detoxication sulphur in 8-weeks-old female Sprague-Dawley (SD) rats. The rats were treated with dose of 0.2%, 1%, 5% detoxication sulphur and 1% sulphur of feed consumption administered for 13 weeks. To evaluate the safety of detoxication sulfur, we examined the body weight, the feed intake, the clinical signs, the ophthalmological test, the hematological and the serum biochemical analysis. We also observed the histopathological changes of liver and kidney in rats. As a result, no significant differences in body weight, feed intake, hematological examination and histopathological between control and detoxication sulphur treatment group were found. Serum biochemical results were not shown significant differences in 0.2% and 1% the treated groups compared with control group. But glucose level were decrease, also ALT and ALP level were increase in 5% treated group. All of these results indicate that 1% detoxication sulphur of feed consumption may be safety in SD rat.

• Journal of Food Hygiene and Safety
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• v.24 no.1
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• pp.69-77
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• 2009

This study was performed to investigate the effect of dried powder of chestnut on lipid metabolism, anti-thrombotic effect in rats. Thirty 5-week-old male Sprague Dawley (SD) rats were randomly allocated into five groups and used for experiment. We examined the lipid metabolism and antithrombotic capacity of SD rats administered for 5 weeks with 0.16 g/kg, 0.5 g/kg chestnut flesh powder and 0.16 g/kg, 0.5 g/kg chestnut inner shell and flesh powder mixture, respectively. Food intake, body weight gain and food efficiency ratio were also checked. The levels of serum triglyceride and tree fatty acid were not statistically significant between the all experimental groups. However, the antithrombotic capacity and total lipid levels of the treatment groups were significantly lower than those of the negative control group. These results suggest that the supplementation of chestnut on diet lower the total lipid level in SD rats.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.146-151
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• 2013

Perilla frutescens usually dieted in East Asian country such as Korea and Japan. Antioxidant, antiinflammatory and anticancer activities of perilla leaves have been founded. In previous study, we confirmed that caffeic acid, major compound of perilla, was accumulation by sucrose aqueous solution and thus antioxidant effect of perilla was enhanced. In this study, we investigated the protective effect of functional perilla leaves extract (PLE) against tert-butyl hydroperoxide(t-BHP) induced-oxidative hepatotoxicity. The pretreatment with PLE (250, 500 and 1000 mg/kg b.w.) for 5 days before a single dose of t-BHP (i.p.; 0.5 mmol/kg) significantly lowered the serum levels of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase dose-dependently. And we confirmed that the indicators of oxidative stress were remarkably reduced in the liver, such as the glutathione contents and malondialdehyde, marker of lipid peroxidation. Pathological histology of the rat livers tissues showed that PLE reduced the hepatocyte degeneration and neutrophilic infiltration of liver induced by t-BHP. These results suggest that functional perilla frutescens has the protective effect of liver against t-BHP-induced oxidative hepatic stress in rats.

• Korean Journal of Food Science and Technology
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• v.26 no.3
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• pp.213-220
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• 1994

Gastrodia (G) Rhizoma has been used clinically as an oriental herbal medicine with sedative, anticonvulsive, and depressor effects. The present study tested effects of G. Rhizoma extracts on the coronary circulation and myocardial oxygen consumption in perfused rat hearts. Sprague Dawley rats (SD) and spontaneously hypertensive rats (SHR) were employed as experimental animals and nonworking Langendorff heart perfusion technique introduced for heart experiments. G. Rhizoma extracts were prepared from grinding G. Rhizoma into powder, extracting in water and 50% ethanol for 4 or 16 hr and diluting with Krebs-Henseleit bicarbonate perfusion buffer to be 70%. Hearts were perfused with bicarbonate buffer oxygenated with 95% $O_{2}:$ 5% $CO_{2}$ at constant coronary perfusion pressure of $90cmH_{2}O$. The diluted extracts were infused into coronary arteries in a concentration of $1{\sim}5\;{\mu}M$ for $7{\sim}8 min. While in SD water- or ethanol-extracts of G. Rhizoma extracted for 16 hr increased coronary perfusate flow (CPF) and decreased coronary vascular resistance (CVR), ethanol-extracts in SHR produced coronary vasoconstriction associated with enhanced CVR. G. Rhizoma extracts-induced increase in CPF reduced myocardial oxygen extraction, and thus myocardial oxygen consumption ($MVO_{2}$) remained at that observed prior to infusion of extracts. In SD and SHR 16 hr-water-extracts markedly altered coronary venous effluent pH and $Pco_{2}$ and evoked metabolic acidosis, which could be a coronary vasodilator mechanism decreasing CVR. In this study, the extracts decreasing CVR in SD and SHR did not augment the lactate production. Therefore, although the effects of the extracts on cardiac function and coronary circulation depended on solvents and duration for extraction, the 16hr-water-extracts, at least, exhibited coronary vasodilation in SD and SHR. Conversely, ethanol-extracts constricted coronary arteries in SHR. G. Rhizoma extracts-induced vasodilation might be due to the metabolic acidosis rather than due to the increased lactate production. The results indicate that G. Rhizoma extracts obtained from proper extracting procedures can be used as a safe and clinically applicable herbal medicine in the cardiovascular diseases such as coronary artery disease and hypertension for vasodilatory and antihypertensive actions.