• Biomolecules & Therapeutics
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• v.3 no.4
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• pp.266-272
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• 1995

The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently through the use of specific antibodies. To generate and characterize a moloclonal antibody against $\beta$-adrenergic receptor, a synthetic $\beta$2-adrenergic receptor peptide (Phe-Gly-Asn-Phe-Trp-Cys-Phe-Trp-Thr-Ser-lle-Asp-Val-Leu) which may comprise part of $\beta$-adrenergic receptor ligand binding pocket was coupled to Keyhole Limpet Hemocyanin (KLH) and used as an immunogen. Male BALB/C mice were immunized with this antigen and the immunized spleen was fused with myeloma SP2/0-Ag14 cells to produce monoclonal antibodies. Two clones were obtained but one of monoclonal antibodies, mAb5G09, was used throughout in this study because the other clone, mAb5All showed weak immunoreactivity against KLH as well. The mouse monoclonal antibody mAb5G09 produced in this study showed immunoreactivity to peptide-KLH conjugates and also to human A43l cells and guinea pig lung $\beta$2-adrenergic receptor as revealed by ELISA and western blot. In the course of determination of the effects of mAb5G09 on $\beta$-receptor ligand binding, it was observed that mAb5G09 specifically bound $\beta$-adrenergic radioligand [$^3$H]dihydroalprenolol (DHA) with a dissociation constant (Kd) of 60 nM. The [$^3$H]DHA binding activity of mAb5G09 had characteristics of immunoglobulins and the binding activity was not observed in the control anti-KLH monoclonal antibody. The monoclonal antibody, mAb5G09 produced in this study may provide useful models for the study of the structure of receptor binding sites.

• Journal of The Korean Society of Emergency Medicine
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• v.28 no.5
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• pp.457-466
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• 2017

Purpose: Traumatic brain injury (TBI) is a significant cause of morbidity and mortality worldwide. Severity of the initial insult is one of the most significant factors affecting outcome following TBI. In order to investigate the mechanisms of cellular injury and develop novel therapeutic strategies for TBI, we designed a standardized animal TBI model and evaluated histological and functional outcomes according to the degree of impact severity. Methods: Male adult C57Bl/6 mice underwent controlled cortical impact (CCI) at varying depths of deflection (1.0-2.0 mm). We performed hematoxylin and eosin staining at 7 days after recovery from TBI. Neurobehavioral characterization after TBI was analyzed by the Barnes maze test, passive avoidance test, open field test, rotarod test, tail suspension test, and light/dark test. Results: We observed a graded injury response according to the degree of deflection depths tested (diameter, 3 mm; velocity, 3 m/s; and duration, 500 ms) compared to sham controls. In the Barnes maze test, the severe TBI (2 mm depth) group showed reduced spatial memory as compared with the sham and mild TBI (1 mm depth) groups at 7 days after TBI. There was a significant difference in the results of the open field test and light/dark test among the three groups. Conclusion: Our findings demonstrate that the graded injury responses following TBI resulted in differential histopathological and behavioral outcomes in a mouse experimental CCI model. Thus, a model of CCI with histologic/behavioral outcome analysis may offer a reliable and convenient design for preclinical TBI research involving mice.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.3
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• pp.279-286
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• 2005

Objective: Although several genetic factors have been associated with defects in human spermatogenesis, the unambiguous causative genes have not been elucidated. The male infertility by haploinsufficiency of PRM1 or PRM2 has been reported in mouse model. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) of PRM1 and PRM2, related to the genotype of Korean infertile men. Methods: Genomic DNAs were extracted from peripheral bloods of infertile men with oligozoospermia or azoospermia, and analyzed using polymerase chain reaction (PCR) and direct sequencing. We carried out the direct sequencing analysis of amplified fragments in PRM1 (557 nucleotides from -42 to 515) and PRM2 (599 nucleotides from 49 to 648) genes, respectively. Results: Three SNPs of coding region in the PRM1 gene was found in the analysis of 130 infertile men. However, the SNPs at a133g (aa 96.9%, ag 3.1% and gg 0.0%), c160a (cc 99.2%, ca 0.8% and aa 0.0%) and c321a (cc 56.9%, ca 35.4% and aa 7.7%) coded the same amino acids, in terms of silence phenotypes. On the other hand, as results of the PRM2 gene sequencing in 164 infertile men, only two SNPs, g398c (gg 62.2%, gc 31.1% and ga 6.7%) and a473c (aa 63.4%, ac 29.9% and cc 6.7%), were identified in the intron of the PRM2 gene. Conclusions: There was no mutation and significant SNPs on PRM1 and PRM2 gene in Korean infertile men. These results suggest that the PRM1 and PRM2 genes are highly conserved and essential for normal fertility of men.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.4 no.2
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• pp.65-72
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• 2018

Prostate specific membrane antigen (PSMA) is a cell surface membrane protein, which is overexpressed in most prostate cancer. Recently, PET imaging with $[^{68}Ga]$PSMA-HBED-CC has been widely used for the diagnosis of recurrent prostate cancer and the studies on the diagnostic potential of $^{64}Cu$-labeled PSMA ligands reported actively. In this study, we monitored with biological evaluation in vivo and PET imaging of $^{64}Cu$-labeled PSMA ligand ($[^{64}Cu]$PSMA-617). The radiolabelling efficiency and stability of $[^{64}Cu]$PSMA-617 were confirmed by radio-thin layer chromatography. The radiolabeling efficiency of $[^{64}Cu]$PSMA-617 showed over 95%, and stabilities of intact remained over 98% in both human and mouse serum for 48 h. In normal male mice, in vivo uptake of $[^{64}Cu]$PSMA-617 in several organs was measured at 2, 4, 6, 24, 48 h after injection. Rapid blood clearance was observed for $[^{64}Cu]$PSMA-617. The high uptake was observed in the lung, liver, intestines and kidneys at 2 h postinjection, but was low in the other organs (1-2 %ID/g) at 4 h. The dynamic PET/CT images of 22RV1 tumor-bearing nude mice were acquired during 60 min and additionally acquired 24 h and 48 h after injection. In dynamic PET images, $[^{64}Cu]$PSMA-617 uptake ratio in tumors versus muscle was increased as time elaplsed until 60 minutes and remained in tumors at 48 h. In these results, the PET/CT imaging using $[^{64}Cu]$PSMA-617 in prostate cancer is expected to be useful for the diagnosis and treatment of prostate cancer patients.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.196-208
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• 2019

Background: Nonalcoholic steatohepatitis (NASH) is one of the chronic inflammatory liver diseases and a leading cause of advanced liver fibrosis, cirrhosis, and hepatocellular carcinoma. The main purpose of this study was to clarify the effects of GBCK25 fermented by Saccharomyces servazzii GB-07 and pectinase, on NASH severity in mice. Methods: Six-wk-old male mice were fed either a normal diet (ND) or a Western diet (WD) for 12 wks to induce NASH. Each group was orally administered with vehicle or GBCK25 once daily at a dose of 10 mg/kg, 20 mg/kg, 100 mg/kg, 200 mg/kg, or 400 mg/kg during that time. The effects of GBCK25 on cellular damage and inflammation were determined by in vitro experiments. Results: Histopathologic analysis and hepatic/serum biochemical levels revealed that WD-fed mice showed severe steatosis and liver injury compared to ND-fed mice. Such lesions were significantly decreased in the livers of WD-fed mice with GBCK25 administration. Consistently, mRNA expression levels of NASH-related inflammatory-, fibrogenic-, and lipid metabolism-related genes were decreased in the livers of WD-fed mice administered with GBCK25 compared to WD-fed mice. Western blot analysis revealed decreased protein levels of cytochrome P450 2E1 (CYP2E1) with concomitantly reduced activation of c-Jun N-terminal kinase (JNK) in the livers of WD-fed mice administered with GBCK25. Also, decreased cellular damage and inflammation were observed in alpha mouse liver 12 (AML12) cells and RAW264.7 cells, respectively. Conclusion: Administration of GBCK25 ameliorates NASH severity through the modulation of CYP2E1 and its associated JNK-mediated cellular damage. GBCK25 could be a potentially effective prophylactic strategy to prevent metabolic diseases including NASH.

• The Korea Journal of Herbology
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• v.36 no.5
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• pp.101-108
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• 2021

Objectives : A root of Dipsacus asperoides C. Y. Cheng et T. M. Ai (D. asperoides) has been traditionally used as a medicinal resource in several Asian countries, including Korean and traditional Chinese medicine that has been traditionally used for treating several medical conditions including pain, arthritis, and bone fractures in Korea. In the present study, we investigated potential subacute toxicities of D. asperoides extract. Methods : C57BL/6 mice (male, 7weeks) were randomly divided into 4 groups of 5 mice. Except for the control group, the mice were orally administrated D. asperoides extract at doses of 50, 150, or 450 mg/kg/day for 2 weeks. At the end of the treatment period, all mice were euthanized, and the following parameters were examined: mortality, body weight, clinical signs, gross findings, hematology, serum biochemistry, organ weight, and histopathology. Results : There were no abnormalities in mortality, clinical signs, body weight, gross findings, or organ weight after repeated administration of D. asperoides extract for 2 weeks, compared with the control group. In addition, there were no significant changes in hematological, serum biochemical, and histopathological parameters between the control group and D. asperoides extract administrated groups with doses of up to 450 mg/kg/day. Conclusion : In this study, D. asperoides extract showed no significant toxicities at a dose of up to 450 mg/kg/day in mice. Although we could not confirm the toxic dose of D. asperoides extract, it can be considered safe for further pharmacological use.

• Journal of Nutrition and Health
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• v.53 no.6
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• pp.570-582
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• 2020

Purpose: Obesity is a risk factor for various adult diseases such as type 2 diabetes, cardio-cerebrovascular disease, and cancer. With an increasing obesity population worldwide, the prevention of obesity with natural components has emerged as an alternative health care strategy. Ramulus mori (Sangzhi, RM) is widely used as a traditional herbal medicine in East Asia. It contains various phytochemicals, including stilbenes and 2-arylbenzofurans. In this study, we compared the anti-obesity effects of RM extracts and its major stilbene components (mulberroside A [MSA] and oxyresveratrol [ORT]) in high fat diet (HFD)-fed obese mice. Methods: Five week-old, male C57BL/6J mice were grouped into 7 experimental groups: normal diet (ND), HFD, HFD + 1% RM water extracts (MW), HFD + 0.1% MSA, HFD + 1% RM ethanol extracts (ME), HFD + 0.1% ORT, and HFD + 1% Garcinia cambogia extracts (GC) as a positive control. All mice were fed experimental diet for 13 weeks. Results: Compared to the HFD group, total body weight and weekly body weight gain were significantly decreased in the ME, ORT, and GC groups. Glucose tolerance level was significantly decreased in all experimental groups, whereas plasma insulin level was decreased in MSA, ME, ORT and GC groups. Plasma glucose, triglyceride (TG), and total cholesterol levels were significantly decreased, whereas high-density lipoprotein cholesterol levels were increased in the MSA, ORT, and GC groups. Hepatic TG accumulation was also significantly decreased in the MSA, ME, ORT, and GC groups. Adipose tissue weight and size of adipocytes were significantly decreased in the MSA, ME and ORT groups, and were comparable to values obtained in the GC group. The levels of adiponectin and SREBP1c mRNA expressions were increased in the ORT and GC groups. Conclusion: These results indicate that ME, ORT and MSA exert significant anti-obesity effect, and have the potential to be developed as a weight control ingredient of functional foods.

• The Korea Journal of Herbology
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• v.37 no.3
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• pp.29-39
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• 2022

Objective : This study is aimed to evaluate the protective effects of Rehmanniae Radix, Mori Folium, and Liriopie Tuber mixture (RML) on lung injury of Particulate matter less than 10 um in diameter and diesel exhaust particles (PM10D) mice model. Methods : To investigate the anti-inflammatory activity of RML, PM10D was diluted in aluminum hydroxide (Alum) in 7-week-old male mice and induced by Intra-Nazal-Tracheal (INT) injection method. Animal experiments were divided into 5 groups. Nor (normal mice), CTL (PM10D-induced mice with the administration of distilled water), DEXA (PM10D-induced mice with the administration of 3 mg/kg Dexamethasone), RML 100 (PM10D-induced mice treated with RML 100 mg/kg weight), and RML 200 (PM10D-induced mice treated with RML 200 mg/kg body weight). After 11 days administration, mice were sacrificed and inflammation-related immune cells in broncho-alveolar lavage fluid (BALF) were analyzed. Inflammation-related biomarkers were also analyzed in blood and lungs. Lung tissue was observed through histological examination. Results : In the PM10D induced model, the PML showed decreases in CXCL-1 and IL-17A in BALF. Expression of inflammatory cytokines and cough-related mRNA genes was significantly decreased in serum and lung tissue. The mixture treatment of RML significantly improved the immune related cells in the serum. In addition, histological observations showed a tendency to decrease the severity of lung injury. Conclusions : Overall, these results confirmed the respiratory protective effect of the RML mixture in a model of lung injury induced by air pollution (PM10+DEP), suggesting that it is a potential treatment for respiratory damage.

• The Korea Journal of Herbology
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• v.29 no.2
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• pp.7-13
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• 2014

Objectives : Chronic mild stress (CMS) model is currently recognized as a better animal model of depression. The purpose of this study was to investigate the antidepressant-like effects of the Nelumbo nucifera leave extract using CMS model. Methods : The antidepressant-like effects of Nelumbo nucifera leaves extract was determined by using animal models of depression. Male ICR mice were divided into four groups: saline-treated normal, without CMS; saline-treated stress control; CMS+ Imipramine(20mg/kg); CMS+Nelumbo nucifera leaves extract(200mg/kg). All mice except the normal group exposed an unpredicted sequence of chronic mild stressors for 5 weeks. The behavior of mice were detected by sucrose preference test, forced swim test and tail suspension test. Then concentration of corticosterone in serum was detected by enzyme immunoassay. Results : Nelumbo nucifera leaves extract administration by daily gavage from the 3rd week exhibited an antidepressant-like effect on CMS-induced depression in mice. Nelumbo nucifera leaves extract administration at dose of 200 mg/kg significantly increased the sucrose consumption, and decreased the immobility durations in forced swim test and tail suspension test. Furthermore the corticosterone level decreased than control group. Conclusions : Chronic mild stress can affect mouse behavior and corticosterone level and cause depression. The present experiments not only further confirm the antidepressant-like effects of Nelumbo nucifera leaves extract in the sucrose preference test, forced swimming test and tail suspension test, but also the improving effects of Nelumbo nucifera leaves extract on the depression-like symptoms in the CMS model. Nelumbo nucifera leaves extract has the antagonism on CMS and produce antidepressive effects.

• The Korea Journal of Herbology
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• v.33 no.6
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• pp.35-42
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• 2018

Objectives : The aim of this study was to investigate whether the extract of Coptidis Rhizoma inhibits inflammation in chronic cold stress (CCS)-exposed mice or not. Methods : Coptidis Rhizoma extract (CRE) was made by reflux with distilled water. Male ICR mice (7 weeks old) were divided randomly into 5 groups: (1) control, (2) CCS, (3) CCS+CRE 100 mg/kg, (4) CCS+CRE 300 mg/kg, (5) CCS+CRE 1,000 mg/kg groups. Mice were orally administered once a day for 14 days starting from 1 day before CCS. Group (2)-(5) were exposed to CCS conditions that maintained at $4^{\circ}C$ for 2 h once a day for 14 days. The levels of serum cortisol and hypothalamic prostaglandin E1 (PGE1) and PGE2 were measured by enzyme-linked immunosorbent assay kit. The expression levels of several pro-inflammatory factors like heat shock protein 70 (HSP70), c-fos, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) were measured by western blot analysis in mouse hypothalamus. Results : Oral administration of CRE 1,000 mg/kg significantly suppressed the increase of serum cortisol levels in mice exposed to CCS. CCS-exposed mice had significantly increased the expression of HSP70, c-fos, and NF-kB in hypothalamus, while CRE treatment significantly attenuated the elevation of these pro-inflammatory factors. The ratio of PGE2/PGE1 was also higher in CCS-exposed mice than control group. CRE treatment significantly reduced the increase of PGE2/PGE1 ratio induced by CCS. Conclusion : These findings suggest that Coptidis Rhizoma may work as a potential agent to modulate inflammatory responses under the condition of cold adaptation formed by CCS.