• The Korean Journal of Zoology
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• v.33 no.2
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• pp.183-190
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• 1990

The present study exarnines the physiological alteradons in prolactin (PRL) messenger ribonucleic acid (mRNA) and serum PRL levels during the rat estrous cycle and the effed of naloxone, an endogenous oploid peptide receptor antagonist, on PRL gene expression during the rat estrous cycle. Adult female rats exhibiting at least two consecutive 4-day estrous cycles were used in this study. A single injection of naloxone (2mg/kg b.w.) or saline was given sc 30 mm prior to decapitation. Animals were sacrificed at 10:00 h of each stage of the estrous cycle, and at 2-h intervals from 10:00 h to 20:00 h during the proestrus. PRL mRNA and serum PRL levels were determined by a RNA-blot hybridization with the rat PRL cDNA probe and by a PRL radjoimmunoassay, respectively. PRL mRNA and serum PRL levels were not dramatically altered in the morning of each stage of diestrus I, II and proestrus, and naloxone failed to modify the two parameters. During estrus naloxone clearly suppressed serum PRL levels, but it was unable to modify PRL mRNA levels. A more detailed examination of the proestrus stage revealed that PRL mRNA and serum PRL levels were fluctuated as a function of time: PRL mRNA levels reached a maximum level at 12:00 h and gradually decreased until 18:00 h. PRL mRNA levels then rose at 20:00 h. No difference of PRL mRNA levels between the control and naloxone-treated groups was observed. Changes in serum PRL levek during proestrus were conversely related to changes in PRL mRNA: serum PRL levels were low from 10:00 h to 14:00 h, then increased and reached a maximum level at 16:00-18:00 h. Following then, serum PRL levels were decreased. Naloxone was effective in suppressing the charaderistic afternoon surge of PRL from 16:00 h to 20:00 h. These data clearly showed that alterations in PRL mRNA levels were conversely correlated with changes mn serum PRL levels on proestrus, indicating a differential regulation of PRL gene expression and secretion.

• Journal of Pharmacopuncture
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• v.3 no.2
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• pp.169-189
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• 2000

To investigate effects of vanous herbal-acupuncture, which were Daebangpungtang, Oyaksungisan, Dokhwalgisaengtang, and Binsosan on Adjuvant Arthritis in rats, the edema rate, the number of WBC, the quantity of TNF-${\alpa}$, COX -2, and IL-6, and histological test of the muscular tissue were measured in the arthritis part. After elicilating arthritis of Sprague Dawely(SD) rats by injection of Freund's complets adjuvant for 2 weeks, saline was injected for the control group, Daebangpungtang acua-acupuncture was injected for the Daebangpungtang group, Oyaksungisan acua-acupuncture was injected for the Oyaksungisan group, Dokhwalgisaengtang acua-acupuncture was injected for the Dokhwalgisaengtang group, and Binsosan acua-acupuncture was injected for the Binsosan group during 30days. Selected point was ST35 in all groups. 1. The number of WBC was $8.24{\pm}0.51$($10^3$/ml) in the normal group and $27.35{\pm}3.51$($10^3$/ml) in the control group. the Daebangpungtang group and the Oyaksungisan group decreased each $20.42{\pm}2.75$($10^3$/ml) and $19.78{\pm}4.99$($10^3$/ml) in the Exp.ll group. This fact showed that the group Exp.ll was more effective than the control group effectively. (P<0.05) 2. The volume of the paw were checked. The volume of paw was $0.62{\pm}0.11mm in the control group and $0.45{\pm}0.08$mm in the Daebangpungtang group and $0.47{\pm}0.07$mm in the Oyaksungisan group, the swelling of the paw was restricted significantly in the Daebangpungtang and Oyaksungisan group(P<0.05) 3. The bands of the TNF-${\alpa}$, COX -2 in the muscular tissue of the control group wcre certain and thick than other groups(except normal group), and the band of the IL-6 in the muscular tissue of all group(except normal group) were similar forms. 4. In histological finding, because of severe inflammatory reaction, remarkably irregular tissue and large amount of inflammatory cells were found in thc control and Dokhwalgisaengtang group. But the Oyaksungisan and Daebangpul1gtang group showed small amount of inflammatory cells, the refrained inflammatory state and even recovering state. From these results, it is showed Oyaksugisan and Daebangpungtang acua-acupuncturc refrain inflammatory reaction and muscular tissue necrosis in SD rats paw were induced by Freund's complete adjuvant

• Journal of Pharmacopuncture
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• v.16 no.4
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• pp.22-29
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• 2013

Objectives: This study was performed to analyze the toxicity of the test substance, anti-inflammatory pharmacopuncture (AIP), when used as a single intramuscular-dose in 6-week-old, male and female Sprague-Dawley rats and to find the lethal dose. Methods: The experiment was conducted at Biotoxtech according to Good Laboratory Practices. Twenty (20) female and 20 male Spague-Dawley rats were divided into 4 groups of five 5 female and 5 male animals per group. The rats in the three experimental groups received single intramuscular injections with 0.1-$m{\ell}$, 0.5-$m{\ell}$ and 1.0-$m{\ell}$/animal doses of AIP, Groups 2, 3, and 4, respectively, and the control group, Group 1, received a single intramuscular injection with a 1.0-$m{\ell}$ dose of normal saline. Clinical signs were observed and body weight measurements were carried out for 14 days following the injections. At the end of the observation period, hematology, clinical chemistry, histopathological tests and necropsy were performed on the injected parts. Results: No deaths occurred in any of the groups. Also, histopathological tests showed that AIP had no effect on the injected parts in terms of clinical signs, body weight, hematology, clinical chemistry, and necropsy. Conclusions: As a result of single intramuscular-dose tests of the test substance AIP in 4 groups of rats, the lethal dose for both males and females exceeded $1.0m{\ell}$/animal. Therefore, AIP is a relatively safe pharmacopuncture that can be used for treatment, but further studies should be performed.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.6
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• pp.1483-1490
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• 2007

This study was carried out to investigate the effects of GCP treatment on the expression of NOS, c-fos, serotonin and substance P in central nerve system of monosodium iodoacetate(MIA)-induced osteoarthritic pain model. Arthritis was induced by injection of MIA(0.5 mg) into knee joint cavities of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken distilled water for 20 days. Treated group was taken extracts of GCP by oraly for same duration. Normal group(n=8) was infected with normal saline and was taken distilled water for 20 days. The numbers of NADPH-d positive cells in superficial dorsal horn of spinal cord of treated group($21{\pm}5$) was significantly (p<0.01) decreased compared with control($33{\pm}5$). The numbers of NADPH-d positive cell in dorsolateral periaqueductal gray matter of treated group($111{\pm}16$) was significantly(p<0.01) decreased compared with control($143{\pm}14$). The numbers of c-fos positive cells in dorsal periaqueductal gray matter of treated group($57{\pm}16$) was significantly(p<0.01) decreased compared with control($78{\pm}13$). The numbers of c-fos positive cells in paraventricular thalamic nucleus of treated group($60{\pm}15$) was significantly decreased compared with control($88{\pm}27$). The numbers of serotonin positive cells in median raphe nucleus of treated group($171{\pm}31$) was significantly(p<0.05) decreased compared with control($217{\pm}48$). On the basis of these results, we concluded that GCP treatment has inhibiting effects on the pain transmission in monosodium iodoacetate-induced osteoarthritic pain model in rat.

• Journal of Pharmacopuncture
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• v.18 no.1
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• pp.63-71
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• 2015

Objectives: This study was carried out to analyze the single-dose toxicity of Eun-Bi San pharmacopuncture injected into the muscle of Sprague-Dawley (SD) rats. Methods: All experiments were performed at Biotoxtech, an institution certified to conduct non-clinical studies under the Good Laboratory Practice (GLP) regulations. Six week old SD rats reared by ORIENTBIO were chosen for this pilot study. The reason SD rats were chosen is that they have been widely used in safety tests in the field of medicine, so the results can be easily compared with many other databases. The Eun-Bi San pharmacopuncture was made in a clean room at the Korean Pharmacopuncture Institute (KPI, K-GMP). The constituents of the Eun-Bi San pharmacopuncture are Angelicae gigantis radix, Strychni semen and Glycyrrhizae radix. These were extracted at low temperature and low pressure in an aseptic room at the KPI. Doses of Eun-Bi San pharmacopuncture, 0.25, 0.5 and 1.0 mL, were administered to the experimental groups, and a dose of normal saline solution, 1.0 mL, was administered to the control group. This study was performed under the approval of the Institutional Animal Ethics Committee of Biotoxtech Co., Ltd. Results: No deaths or abnormalities occurred in any of the four groups. No significant changes in weight, hematological parameters or clinical chemistry between the control group and the experimental groups were observed. To determine if abnormalities existed in any organs and tissues, we used microscopy to examine representative histological sections of each specified organ; the results showed no significant differences in any of the organs or tissues. Conclusion: The results of this study suggest that treatment with Eun-Bi San pharmacopuncture is relatively safe and that its clinical use may be beneficial. Further evaluations and studies on this subject will be needed to provide more concrete evidence in support of these conclusions.

• Journal of Pharmacopuncture
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• v.18 no.1
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• pp.36-43
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• 2015

Objectives: Water-soluble carthami flos (WCF) is a new mixture of Carthami flos (CF) pharmacopuncture. We conducted a 4-week toxicity test of repeated intramuscular injections of WCF in Sprague-Dawley rats. Methods: Forty male and 40 female rats were divided into 4 groups of 10 male and 10 female SD rats: The control group received 0.5 mL/animal/day of normal saline whereas the three experimental groups received WCF at doses of 0.125, 0.25, and 0.5 mL/animal/day, respectively. For 4 weeks, the solutions were injected into the femoral muscle of the rats alternating from side to side. Clinical signs, body weights, and food consumption were observed; opthalmological examinations and urinalyses were performed. On day 29, blood samples were taken for hematological and clinical chemistry analyses. Then, necropsy was conducted in all animals to observe weights and external and histopathological changes in the bodily organs. All data were tested using a statistical analysis system (SAS). Results: No deaths were observed. Temporary irregular respiration was observed in male rats of the experimental group for the first 10 days. Body weights, food consumptions, opthalmological examinations, urinalyses, clinical chemistry analyses, organ weights and necropsy produced no findings with toxicological meaning. In the hematological analysis, delay of prothrombin time (PT) was observed in male rats of the 0.25- and the 0.5-mL/animal/day groups. In the histopathological test, a dose-dependent inflammatory cell infiltration into the fascia and panniculitis in perimuscular tissues was observed in all animals of the experimental groups. However, those symptoms were limited to local injection points. No toxicological meanings, except localized changes, were noted. Conclusion: WCF solution has no significant toxicological meaning, but does produce localized symptoms. No observed adverse effect level (NOAEL) of WCF in male and female rats is expected for doses over 0.5 mL/animal/day.

• Development and Reproduction
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• v.14 no.3
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• pp.199-205
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• 2010

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD). The present study was undertaken to examine whether depletion of brain dopamine (DA) stores with 6-OHDA can make alteration in the activities of the testicular steroidogenesis in adult rats. Young adult male rats (3 months old) were received a single dose of 6-OHDA (200 ${\mu}g$ in 10 ${\mu}{\ell}$/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. The mRNA levels of steroidogenesis-related enzymes were measured by qRT-PCRs. Serum testosterone levels were measured by radioimmunoassay. Single icv infusion of 6-OHDA significantly decreased the mRNA levels of CYP11A1 (control:6-OHDA group=$1:0.68{\pm}0.14$ AU, p<0.05), CYP17 (control:6-OHDA group=$1:0.72{\pm}0.13$ AU, p<0.05). There were no changes in the mRNA levels of $3{\beta}$-HSD (control:6-OHDA group=$1:0.84{\pm}0.08$ AU) and $17{\beta}$-HSD (control: 6-OHDA group=$1:0.63{\pm}0.20$ AU), though the levels tended to be decreased in the 6-OHDA treated group. Administration of 6-OHDA decreased significantly the mRNA level of StAR when compared to the level of saline-injected control animals (control:6-OHDA group=$1:0.72{\pm}0.08$ AU, p<0.05). Treatment with single dose of 6-OHDA remarkably lowered serum testosterone levels compared to the levels of control group (control:6-OHDA group=$0.72{\pm}0.24:0.13{\pm}0.03ng/m{\ell}$, p<0.05). Taken together with our previous study, the present study demonstrated that the activities of hypothalamus-pituitary-testis hormonal axis could be negatively affected by blockade of brain DA biosynthesis, and suggested the reduced reproductive potential might be resulted in the animals. More precise information on the testicular steroidogenic activities in PD patients and PD-like animals should be required prior to the generalization of the sex steroid hormone therapy to meet the highest standards for safety and efficacy.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.201-206
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• 2006

This study was carried cut to determine the immunological response of uterus-induced by Lipopolysaccharides (LPS) in Holstein cows. The LPS isolated from Bacteroids helcogenes and Fusobacterium varium was injected at the rate of 100 ${\mu}g$ with 30 ml of phospahte buffer saline(PBS) in each cow(n=5). Three cows were acted as control. There was no difference in total polymorphonuclear leukocytes(PMNL) concentration in uterine fluid between control and LPS groups at 24, 48 and 72 hrs after LPS treatment. There was significant difference in rate of PMNL between control and LPS groups at 24(41.7% vs 72.1%), 48(41.0% vs 81.6%) and 72 hrs(44.3% vs 79.0%) after LPS treatment. There was no difference in PMNL viability between control and LPS groups at 24, 48 and 72 hrs after LPS treatment. There was significant difference in rate of phagocytic PMNL between control and LPS groups at 48 hr after LPS treatment(1.1% vs 7.7%).

• Journal of radiological science and technology
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• v.35 no.2
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• pp.151-156
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• 2012

$^{18}F$-fluorodeoxyglucose ($^{18}F$-FDG) causes a significant amount of radioactivity retention in kidneys and urinary tract and degrades image quality and diagnostic performance. Diuretics are used to perform tests and prevent the urinary tract retention of $^{18}F$-FDG. The purpose of the study is to investigate how the diuretics affect images and excretion rates of $^{18}F$-FDG. The study consists of a group using diuretics for patients with no primary tumors or transfer lesions in kidneys according to PET/CT images, a group using physiological saline and the control group injecting only $^{18}F$-FDG and SUVs are measured by configuring interested areas for each group. Also, SUVs are compared and evaluated depending on the lasix injection after basic inspection and injecting $^{18}F$-FDG for quantitative analysis. The study shows that images with decreased background radioactivity and increased urine excretion due to using diuretics. However, an opposite result that there is no change in the amount of radioactivity in urine appears. The study concludes that the diuretics may decrease background radioactivity in the images but may not affect the $^{18}F$-FDG excretion.

• Childhood Kidney Diseases
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• v.14 no.1
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• pp.32-41
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• 2010

Purpose : Glomerulonephropathy (GN) often manifests as proteinuria and progresses to chronic renal failure without specific therapy. Mesenchymal stem cell (MSC) has been tried as a therapeutic agent in experimental GN, and previous studies showed that administration of MSC concomitantly to the insult inducing GN or via intra-renal administration ameliorated proteinuria. The purpose of this study was to test the therapeutic potential of MSC administered via intravenous route at the time of clinically evident proteinuria. Methods : MSCs were administered intravenously via tail vain into the mice with adriamycin (ADR) induced nephropathy (ADR-GN), two weeks after ADR injection when massive proteinuria was evident. To test the capacity of MSC modulate the cytokine production in the inflammatory milieu, the concentrations of IFN-$\gamma$ and IL-10 were measured in the supernatant of in vitro mixed lymphocyte culture (MLC) with or without additional MSC. Results : MSCs administered intravenously into the proteinuric mice with ADR-GN accelerated the recovery of this experimental GN with disappearance of proteinuria in two weeks when the saline treated (control) mice still showed significant proteinuria. The mice treated with MSC also had a tendency of better survival. Addition of MSC decreased IFN-$\gamma$ and increased IL-10 in the supernatant of MLC. Conclusion : This study showed that MSC had a therapeutic potential even when administered in a more clinically relevant setting into a proteinuric glomerulonephropathy model. Further study to verify the mechanism and long-term safety of this phenomenon is required.