• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.382-389
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• 2017

Currently, reticulocyte experimental calculation technology used in clinical laboratories are divided two types: manual and automated. Manual reticulocyte counting using a microscopy lacks accuracy due particularly to its low reproducibility, affecting the accuracy of manual reticulocyte count. Moreover, Automatic blood corpuscle analyzer flow cytometry is difficult to be used in underdeveloped countries and small scale laboratories due to relatively high cost. Therefore, this study tried to find a new method to complement these drawbacks. The aim of this study was to compare the stained reticulocytes count by spectrophotometer and also to analyze the statistics of spectrophotometer and flow cytometer. The same 8 EDTA samples were repeated 36 times to compare the agreement between spectrophotometer and flow cytometer. This study measured the specimen diluted 600 times at 700~780 nm by 10 differences. Wavelength between 710 to 730 by absorbance showed a positive correlation between standard data and test data (r=0.967, p<0.01), presenting a correlation between variables. Statistical analyses of regression for test and standard parametric data, the optimal dilution factor was 600 times. Therefore, this study tried to technical utilizes such as contributing economical for the reticulocyte absorbance apply from the auto spectrophotometer, a monitoring system for the reticulocyte relation anemia, etc. Therefore, more extensive studies, including an auto chemical analyzer application, will be needed.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.2
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• pp.357-364
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• 2008

Mylabris is the dried body of the chinese blister beetle. The species used in medicine are Mylabris phalerata and M. cichorii. In recent studies, it has been found that Mylabris possesses antitumor properties, increases the number of leukocytes, and has irritant effects on the urinary organs. The crude extracts of Mylabris have been noted for their highly irritant action and other traditional uses of Mylabris include treatment of poor local blood circulation. The active constituent of Mylabris is cantharidin. The chemical is notable for its vesicant properties, but with severe side effects such as nephrotoxicity. This experiment examined the effect of extracts and fractions, obtained from Mylabris phalerata Pall. on hair growth activity of the C57BL/6N mice after topical application to skin. First, we examined the effect of an extracts, obtained from the alcohol extracts of dried Mylabris phalerata Pall. on hair growth activity of the C57BL/6N mice after topical application to skin. Second, we examined on hair growth activity of the cantharidin fraction of Mylabris phalerata Pall. compared to the control and 1% minoxidil groups. Third, we investigated the number of hair follicle and mast cells after topical application of extracts of Mylabris phalerata Pall. to skin for 16 days. The results were as follows: Hair growth effect from the extracts of Mylabris phalerata Pall.(0.312%) was observed in 80% of mice whose hair had been removed in 13 days. Hair growth effect from the extract of Mylabris phalerata Pall.(0.312 and 0.625%) and 1% minoxidil group was observed in 100% of mice whose hair had been clipped in 20 days. Hair growth effect from the cantharidin fraction(0.5%) and water fraction(0.5%) of Mylabris phalerata Pall. was observed in 100% of mice whose hair had been clipped in 24 days. The hair growth effect on the cantharidin fraction(0.125%) was observed to be strong compared with the minoxidil(3%) group, commercial hair growth agents, in mice whose hair had been clipped in 19 days. In the spontaneous alopecia mice model, the hair growth effect from the cantharidin fraction (0.125%) was observed to be strong as compared with the states before the 13 days experiment. These experiments suggest that extracts and fractions of Mylabris phalerata Pall. may stimulate the topical hair growth activity in low doses.

• Molecular & Cellular Toxicology
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• v.4 no.4
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• pp.348-354
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• 2008

Bisphenol A (BPA), an environmental endocrine disrupter, enters the human body continuously in food and drink. Young children are likely to be more vulnerable than adults to chemical exposure due to the immaturities of their organ systems, rapid physical development, and higher ventilation, metabolic rates, and activity levels. The direct effect of BPA on peripheral tissue might also be of importance to the development of insulin resistance. However, the influence that BPA has on insulin signaling molecules in skeletal muscle has not been previously investigated. In this study, we examined the effect of BPA on fasting blood glucose (FBG) in post-weaned Wistar rats and on insulin signaling proteins in C2C12 skeletal muscle cells. Subsequently, we investigated the effects of BPA on insulin-mediated Akt phosphorylation in C2C12 myotubes. In rats, BPA treatment (0.1-1,000 ng/mL for 24 hours) resulted in the increase of FBG and plasma insulin levels, and reduced insulin-mediated Akt phosphorylation. Furthermore, the mRNA expression of insulin receptor (IR) was decreased after 24 hours of BPA treatment in C2C12 cells in a dose-dependent manner, whereas the mRNA levels of other insulin signaling proteins, including insulin receptor substrate-1 (IRS-1) and 5'-AMP-dependent protein kinase (AMPK), were unaffected. Treatment with BPA increased GLUT4 expression and protein tyrosine phosphatase 1B (PTP1B) activity in C2C12 myotubes, but not in protein levels. We conclude that exposure to BPA can induce insulin resistance by decreasing IR gene expression, which is followed by a decrease in insulin- mediated Akt activation and increased PTP1B activity.

• Food Science of Animal Resources
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• v.30 no.6
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• pp.938-945
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• 2010

This study was conducted to investigate the dietary effects of germinated and fermented (with Monascus pupureus) soybean screenings (GFS) on growth performance and meat quality in broiler chicken. A total of 750 1-day-old Ross ${\times}$ Ross male broiler chicks were randomly allocated into five groups (five replications with 30 birds each) and fed experimental diets for 5 wks as follows: Group 1, negative-control (antibiotics-free diet); Group 2, positive-control (negative-control with 10 ppm of Avilamycin); Group 3, negative-control with 0.3% GFS; Group 4, negative-control with 0.5% GFS; Group 5, negative-control with 1% GFS. The final body weight of each group fed a diet containing 1% GFS was significantly higher than that of the negative-control group. The feed conversion ratios of all groups fed diets containing GFS and the positivecontrol group were significantly improved compared to the negative-control group during the whole period (p<0.05). The relative weights of various organs along with the activities of serum glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) were not influenced by the dietary treatments. The levels of total serum cholesterol in groups fed diets containing 0.5% and 1% GFS were more significantly lowered compared to those of the control groups without GFS (p<0.05). There were no significant differences in the cecal microflora profiles among the groups. Further, the dietary treatments did not influence the physico-chemical properties of the edible meat, including the shear force, pH, meat color (CIE $L^*$, $a^*$ and $b^*$), and content of malondialdehyde (MDA). Cooking loss of breast muscle in the groups fed GFS was significantly lowered compared to that of the negative control group (p<0.05). These results indicate that dietary GFS could improve growth performance in broiler chicken and may affect meat quality in some instances.

• Archives of Pharmacal Research
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• v.25 no.5
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• pp.655-663
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• 2002

Liver fibrosis is a prepathological state wherein damaged liver tissues in chronic liver diseases, such as hepatitis, are not repaired to normal tissues, but converted to fibrous tissue. 5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione (oltipraz), a cancer chemopreventive agent, is effective against a wide variety of chemical carcinogens. Recently, we reported that oltipraz inhibits liver fibrogenesis (Kang et al., 2002). In the present study, the effects of oltipraz in combination with dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2'-dicarboxylate (DDb) on dimethylnitrosamine (DMN)-induced liver fibrogenesis were assessed in rats. Oltipraz (30 mg/kg body weight, po, 3 times per week for 4 weeks) was found to inhibit the increases in plasma ALT, AST and bilirubin by DMN, whereas DDB (30 mg/kg body weight, po, 3 times per week for 4 weeks) attenuated the increases in the plasma ALT and bilirubin. The lowered plasma protein and albumin contents in DMN-treated rats were completely restored by oltipraz, but not by DDB. DDB decreases liver cell injury and inflammation through inhibition of nuclear factor-kB. DMN increased the accumulation of liver collagen, as indicated by the increase in the 4-hydroxyproline content in liver homogenates, which was reduced by treatment with oltipraz, but not by DDB. Given the differential effect between oltipraz and DDB, the potential enhancement of antifibrotic efficacy by the drugs was assessed in the animal model. Despite the minimal effect of DDB on DMN-induced fibrogenesis, DDB (5-25 mg/kg), administered together with oltipraz (25-5 mg/kg), showed an additive protective effect against hepatotoxicity and fibrosis induced by DMN, which was shown by the blood chemistry parameters and histopathological analysis. The adequate composition ratio of oltipraz to DDB was 5:1. These results provide information on the pharmaceutical composition, comprising of oltipraz and DDB as the active components, for the treatment and/or prevention of liver fibrosis and cirrhosis.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• Molecules and Cells
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• v.19 no.3
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• pp.402-407
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• 2005

Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury, but this approach is limited by their poor viability after transplantation. To reduce cell loss after transplantation, we introduced the fibroblast growth factor-2 (FGF-2) gene ex vivo before transplantation. The isolated MSCs produced colonies with a fibroblast-like morphology in 2 weeks; over 95% expressed CD71, and 28% expressed the cardiomyocyte-specific transcription factor, Nkx2.5, as well as ${\alpha}$-skeletal actin, Nkx2.5, and GATA4. In hypoxic culture, the FGF-2-transfected MSCs (FGF-2-MSCs) secreted increased levels of FGF-2 and displayed a threefold increase in viability, as well as increased expression of the anti-apoptotic gene, Bcl2, and reduced DNA laddering. They had functional adrenergic receptors, like cardiomyocytes, and exposure to norepinephrine led to phosphorylation of ERK1/2. Viable cells persisted 4 weeks after implantation of $5.0{\times}10^5$ FGF-2-MSCs into infarcted myocardia. Expression of cardiac troponin T (CTn T) and a voltage-gated $Ca^{2+}$ channel (CaV2.1) increased, and new blood vessels formed. These data suggest that genetic modification of MSCs before transplantation could be useful for treating myocardial infarction and end-stage cardiac failure.

• Korean Journal of Acupuncture
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• v.17 no.1
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• pp.101-121
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• 2000

The purpose of this morphological studies was to investigate the relation to the meridian, acupoint and nerve. The common locations of the spinal cord and brain projecting to the the gallbladder, GB34 and common peroneal nerve were observed following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV), into the gallbladder, GB34 and common peroneal nerve of the rabbit. After survival times of 96 hours following injection of PRV, the thirty rabbits were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope. The results were as follows: 1. In spinal cord, PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina V, VII, X, intermediolateral nucleus and dorsal nucleus. 2. In medulla oblongata, The PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nucleus, medullary reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus, gigantocellular nucleus, lateral paragigantocellular nucleus, principal sensory trigeminal nucleus and spinal trigeminal nucleus. 3. In Pons, PRV labeled neurons were parabrachial nucleus, Kolliker-Fuse nucleus and cochlear nucleus. 4. In midbrain, PRV labeled neurons were founded in central gray matter and substantia nigra. 5. In diencephalon, PRV labeled neurons were founded in lateral hypothalamic nucleus, suprachiasmatic nucleus and paraventricular hypothalamic nucleus. 6. In cerebral cortex, PRV labeled neuron were founded in hind limb area.This results suggest that PRV labeled common areas of the spinal cord projecting to the gallbladder, GB34 and common peroneal nerve may be first-order neurons related to the somatic sensory, viscero-somatic sensory and symapathetic preganglionic neurons, and PRV labeled common area of the brain may be first, second and third-order neurons response to the movement of smooth muscle in gallbladder and blood vessels.These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory system monitoring the internal environment, including both visceral sensation and various chemical and physical qualities of the bloodstream. The present morphological results provide that gallbladder meridian and acupoint may be related to the central autonomic pathways.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.62-74
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• 1998

In feedlot cattle the abrupt change of diet from roughage to a large quantity of grain for the purpose to improve production often results in increased occurrence of rumen acidosis or acute carbohydrate encouragement enterotoxemia, bloats diarrhea liver abscess and laminitis or robot disease. The common management practice to control these problem is to increase the amount of concentrates in the diet in a stepwise manner until the animals are adapted to a high-grain ration. However this practice requires at least about 3 weeks adaptation period and specially prepared adaptation rations which contain various amount of concentrates. Present experiment was undertaken in order to findout the more simple and rapid adaptation method of cattle to a high grain ration. Nineteen Korean calves aging from four to six month were fed artifical hay (Youngchoun Chuk-Hyup, Korea) which contains 10% of concentrates or alfalfa and rye grass hays for two months and randomly alloted to three experimental groups and two control groups. The experimental group-1 was inoculated by stomach tube for two days with li500 ml/day of ruminal fluid fished from Korean beef cattle that had been previously adapted to a high-energy ration. The experimental group-2 was inoculated by trocalization for two days with the same ruminal fluid. The experimental group-3 was inoculated by trocalization with 1,500 ml/day of bacterial culture which contained 2$\times $10$^{9}$/m1 of Gram-negative bacteria derived from adapted luminal fluid. The two control groups were treated with normal saline solution by the same methods. All animals were fed high-energy ration that contained 80% of grain ad libitum for 30-74 days beginning on the third of the treatment. The effect of the inoculation on the adaptation was observed clinicopathologically with the following results; All of the experimental calves inoculated with the ruminal fluid or Gram-negative bacterial culture derived from adapted cattle did not show any signs of rumen acidosis or other related diseases, while most of the control calves did show diarrhea and bloat and a calf laminitis. The average daily weight gain and feed efficiency of experimental calves were slightly improved compared with control calves. Following the feeding of high-grain rational the pH of the ruminal fluid was lowered in both the experimental and control groups. However severe acidosis with the pH of below 5.0 was observed in only a control group-2. The protozoal number in ruminal fluid was markedly decreased during the high-grain feeding in both the experimental and control calves. However the decrease was mere severe in control calves compared with the experimental calves. The activation of the protozoa were completely disappeared within nine hours at the refrigerator temperature (4"C). No significant differences in heamatological and blood chemical values between the experimental and control calves were recognized. However in one control calf which showed clinically laminitis marked elevations of serum glutamic oxaloacetate transaminase and lactic dehydrogenase activities and a decrease of serum glucose level were observed. From these results it would be concluded the intraruminal transplantation of unadapted calves with the adapted ruminal fluid from cattle previously adapted to a high-energy ration prevents disease problem associated with high-grain feeding and improve weight gain and feed efficiency.ency.

• Toxicological Research
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• v.35 no.4
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• pp.361-369
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• 2019

1,2-Dichloropropane (1,2-DCP) has been used as an industrial solvent and a chemical intermediate, as well as in soil fumigants. Human exposure may occur during its production and industrial use. The target organs of 1,2-DCP are the eyes, respiratory system, liver, kidneys, central nervous system, and skin. Repeated or prolonged contact may cause skin sensitization. In this study, 1,2-DCP was dissolved in corn oil at 0, 2.73, 5.75, and 8.75 mL/kg. The skin of mice treated with 1,2-DCP was investigated using western blotting, hematoxylin and eosin staining, and immunohistochemistry. 1,2-DCP was applied to the dorsal skin and both ears of C57BL/6J mice. The thickness of ears and the epidermis increased significantly following treatment, and the appearance of blood vessels was observed in the dorsal skin. Additionally, the expression of vascular endothelial growth factor, which is tightly associated with neovascularization, increased significantly. The levels of protein kinase-B (PKB), phosphorylated PKB, mammalian target of rapamycin (mTOR), and phosphorylated mTOR, all of which are key components of the phosphoinositide 3-kinase/PKB/mTOR signaling pathway, were also enhanced. Taken together, 1,2-DCP induced angiogenesis in dermatitis through the PI3K/PKB/mTOR pathway in the skin.