• Archives of Pharmacal Research
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• 제28권12호
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• pp.1350-1357
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• 2005

Nuclear factor kappa B (NF-$\kappa$B) regulates the expression of multiple cytokines, chemokines, and cell adhesion molecules that are involved in the pathogenesis of asthma. We investigated the anti-asthmatic effects and the mechanism of action of DA-9201, an extract of the black rice, in a mouse model of asthma. Mice immunized with ovalbumin (OVA) were administered with DA-9201 (30, 100 or 300 mg/kg) or dexamethasone (DEXA, 3 mg/kg) for 2 weeks and challenged with aerosolized OVA during the last 3 days. Anti-asthmatic effects were assessed by means of enhanced pauses, level of total lgE and Th2 cytokines in plasma or bronchoalveolar lavage fluid (BALF), the percentage of eosinophils in BALF, and histopathological examination. The expression of NF-$\kappa$B in nuclear and cytoplasmic fraction and its DNA-binding activity in lung tissues were analyzed by means of Western blotting and electrophoretic gel mobility shift assay (EMSA), respectively. DA-9201 significantly reduced airway hyperrespon-siveness (AHR), total lgE level in plasma and BALF, IL-4, IL-5, and IL-13 levels in BALF, and the percentage of eosinophils in BALF. Tissue inflammation was significantly improved by DA­9201 treatment. In addition, DA-9201 dramatically suppressed the expression of NF-$\kappa$B and its DNA-binding activity. These results suggest that DA-9201 may be useful for the treatment of asthma and its efficacy is related to suppression of NF-$\kappa$B pathway.

• Journal of Microbiology and Biotechnology
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• 제29권9호
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• pp.1444-1452
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• 2019

The conventional prophylactic vaccines for human papillomavirus (HPV) efficiently prevent infection with high-risk HPV types, but they do not promote therapeutic effects against cervical cancer. Previously, we developed HPV16 E7-expressing Lactobacillus casei (L. casei-E7) as a therapeutic vaccine candidate for cervical cancer, which induces antitumor therapeutic effects in a TC-1 murine cancer model. To improve the therapeutic effect of L. casei-E7, we performed co-treatment with poly-gamma-glutamic acid (${\gamma}-PGA$), a safe and edible biomaterial naturally secreted by Bacillus subtilis. We investigated their synergistic effect to improve antitumor efficacy in a murine cancer model. The treatment with ${\gamma}-PGA$ did not show in vitro cytotoxicity against TC-1 tumor cells; however, an enhanced innate immune response including activation of dendritic cells was observed. Mice co-administered with ${\gamma}-PGA$ and L. casei-E7 showed significantly suppressed growth of TC-1 tumor cells and an increased survival rate in TC-1 mouse models compared to those of mice vaccinated with L. casei-E7 alone. The administration of ${\gamma}-PGA$ markedly enhanced the activation of natural killer (NK) cells but did not increase the E7-specific cytolytic activity of $CD8^+$ T lymphocytes in mice vaccinated with L. casei-E7. Overall, our results suggest that oral administration of ${\gamma}-PGA$ induces a synergistic antitumor effect in combination with L. casei-E7.

• Biomolecules & Therapeutics
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• 제31권4호
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• pp.402-410
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• 2023

Long-term administration of levodopa (L-DOPA) to patients with Parkinson's disease (PD) commonly results in involuntary dyskinetic movements, as is known for L-DOPA-induced dyskinesia (LID). 5-Hydroxytryptophan (5-HTP) has recently been shown to alleviate LID; however, no biochemical alterations to aberrant excitatory conditions have been revealed yet. In the present study, we aimed to confirm its anti-dyskinetic effect and to discover the unknown molecular mechanisms of action of 5-HTP in LID. We made an LID-induced mouse model through chronic L-DOPA treatment to 6-hydroxydopamine-induced hemi-parkinsonian mice and then administered 5-HTP 60 mg/kg for 15 days orally to LID-induced mice. In addition, we performed behavioral tests and analyzed the histological alterations in the lesioned part of the striatum (ST). Our results showed that 5-HTP significantly suppressed all types of dyskinetic movements (axial, limb, orolingual and locomotive) and its effects were similar to those of amantadine, the only approved drug by Food and Drug Administration. Moreover, 5-HTP did not affect the efficacy of L-DOPA on PD motor manifestations. From a molecular perspective, 5-HTP treatment significantly decreased phosphorylated CREB and ΔFosB expression, commonly known as downstream factors, increased in LID conditions. Furthermore, we found that the effects of 5-HTP were not mediated by dopamine1 receptor (D1)/DARPP32/ERK signaling, but regulated by AKT/mTOR/S6K signaling, which showed different mechanisms with amantadine in the denervated ST. Taken together, 5-HTP alleviates LID by regulating the hyperactivated striatal AKT/mTOR/S6K and CREB/ΔFosB signaling.

• Journal of Veterinary Science
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• 제22권2호
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• pp.18.1-18.12
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• 2021

Background: We previously elucidated the protective mechanism of Korean red ginseng oil (RGO) against Brucella abortus infection, and our phytochemical analysis revealed that palmitic acid (PA) was an abundant component of RGO. Consequently, we investigated the contribution of PA against B. abortus. Objectives: We aimed to investigate the efficacy of PA against B. abortus infection using a murine cell line and a murine model. Methods: Cell viability, bactericidal, internalization, and intracellular replication, western blot, nitric oxide (NO), and superoxide (O₂^-) analyses and flow cytometry were performed to determine the effects of PA on the progression of B. abortus infection in macrophages. Flow cytometry for cytokine analysis of serum samples and bacterial counts from the spleens were performed to determine the effect of PA in a mouse model. Results: PA did not affect the growth of B. abortus. PA treatment in macrophages did not change B. abortus uptake but it did attenuate the intracellular survivability of B. abortus. Incubation of cells with PA resulted in a modest increase in sirtuin 1 (SIRT1) expression. Compared to control cells, reduced nitrite accumulation, augmented O₂^-, and enhanced pro-inflammatory cytokine production were observed in PA-treated B. abortus-infected cells. Mice orally treated with PA displayed a decreased serum interleukin-10 level and enhanced bacterial resistance. Conclusions: Our results suggest that PA participates in the control of B. abortus within murine macrophages, and the in vivo study results confirm its efficacy against the infection. However, further investigations are encouraged to completely characterize the mechanisms involved in the inhibition of B. abortus infection by fatty acids.

• Molecules and Cells
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• 제46권12호
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• pp.764-777
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• 2023

Recombinant immunotoxins (RITs) are fusion proteins consisting of a targeting domain linked to a toxin, offering a highly specific therapeutic strategy for cancer treatment. In this study, we engineered and characterized RITs aimed at mesothelin, a cell surface glycoprotein overexpressed in various malignancies. Through an extensive screening of a large nanobody library, four mesothelin-specific nanobodies were selected and genetically fused to a truncated Pseudomonas exotoxin (PE24B). Various optimizations, including the incorporation of furin cleavage sites, maltose-binding protein tags, and tobacco etch virus protease cleavage sites, were implemented to improve protein expression, solubility, and purification. The RITs were successfully overexpressed in Escherichia coli, achieving high solubility and purity post-purification. In vitro cytotoxicity assays on gastric carcinoma cell lines NCI-N87 and AGS revealed that Meso(Nb2)-PE24B demonstrated the highest cytotoxic efficacy, warranting further characterization. This RIT also displayed selective binding to human and monkey mesothelins but not to mouse mesothelin. The competitive binding assays between different RIT constructs revealed significant alterations in IC₅₀ values, emphasizing the importance of nanobody specificity. Finally, a modification in the endoplasmic reticulum retention signal at the C-terminus further augmented its cytotoxic activity. Our findings offer valuable insights into the design and optimization of RITs, showcasing the potential of Meso(Nb2)-PE24B as a promising therapeutic candidate for targeted cancer treatment.

• 한국염색가공학회지
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• 제24권3호
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• pp.196-203
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• 2012

The purpose of this study was to investigate usefulness of the pine needles extract on Atopic Dermatitis(AD)-like skin lesions. To investigate the effect of pine needles extract in vivo, atopic dermatitis (NC/Nga) mice using DNCB (2.4-Dinitrochlorbenzene) was used. NC/Nga mice were challenged with DNCB during 2 weeks to develope AD-like skin lesion. After that, pine needles extract was applied to AD-like skin lesion on the backs of the NC/Nga mice during 3 weeks. The efficacy of pine needles extract in the NC/Nga mice was evaluated by measurement of the skin lesion severity(NC mouse score), the serum IgE level, epidermal thickness changes, and mast cell number. Blood was collected from the retro-orbital area and the level of IgE in the blood was measured. The epidermal thickness and mast cell number were observed by microscopic method after H&E stain. The serum IgE levels were decreased after treatment with pine needles extract. The epidermal thickness and mast cell number were decreased after treatment with pine needles extract. To conclude, the topical application of pine needles extract suppressed the progression of AD-like skin lesion.

• 한국염색가공학회지
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• 제24권3호
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• pp.189-195
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• 2012

The purpose of this study was to investigate the possibility of bamboo extract as natural dye having the effect on atopic dermatitis(AD). To investigate the effect of bamboo extract on AD in vivo, we applied bamboo extract to the AD-like skin lesion the backs of atopic of NC/Nga mice, an animal model of AD. NC/Nga mice were challenged with DNCB(2.4-Dinitrochlorbenzene) to develope AD-like skin lesions. The efficacy of bamboo extract in the NC/Nga mice was evaluated by measurement of the skin lesion severity(NC mouse score), the serum IgE level, epidermal thickness changes, and mast cell number. Bamboo extracts improved skin lesions in NC/Nga mice. The serum IgE levels were decreased after treatment with bamboo extract. Histological examinations revealed a decrease in epidermal thickness and mast cell number after treatment with bamboo extract. To conclude, the topical application of bamboo extract suppressed the progression of AD-like skin lesions.

• 한국한의학연구원논문집
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• 제6권1호
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• pp.99-105
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• 2000

Lithium, a simple monovalent cation, is the mainstay in the treatment of manic-depressive illness, but despite extensive research, its mechanism of action remains to be elucidated. Because lithium requires chronic administration for therapeutic efficacy and because its beneficial effects last well beyond its discontinuation, it has been postulated that lithium may exert major effects at the genomic level. We have previously shown that Yungmijihwangwon (YM) increase TH protein expression through antioxidant activity and unknown effects in vivo. In the pressent study, we have sought to determine if lithium in herbal medicine also increases the expression of endogenous gene known to be regulated by AP-1 and have therefore investigated the effects of herbal medicine on tyrosine hydroxylase (TH) levels. Male mice were treated with LiCl and herbal medicine for 30 days, and TH levels and dopamine level were measured in striatum using immunoblotting and ECD-HPLC. Herbal medicine treatment resulted in statistically nonsignificant increase in TH levels in mouse striatum. The precise therapeutic relevance of these effects is presently unknown.

• 혜화의학회지
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• 제25권1호
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• pp.99-108
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• 2016

While axons in the peripheral nerve can regenerate and lead to functional recovery to a certain extent after injury, its efficacy varies depending on the severity and duration of the injury. Here, we investigated the effects of Boyanghwano-tang (BYHOT) treatment on the regenerative responses in the sciatic nerves after prolonged transection and coaptation surgery. In mice given crush injury, axonal regeneration was completed when analyzed 1 week later and did not show any difference in regenerative reponses in the distal portion of the nerve between saline- and BYHOT-treated groups. In animal models with transection and reconnection, axonal regeneration was markedly retarded compared to animals with crush injury. Regenerating axons were extended into the reconnected distal portion of the nerve more actively in animals treated with BYHOT than saline controls. Cdc2 protein was similarly induced in nerves with crush injury and with transection and recollection, and its level was lower in BYHOT-treated animal than saline control when measured 2 weeks after nerve reconnection. These results suggest that BYHOT may be useful to promote axonal regeneration in the peripheral nerve after severe injury.

• 대한유전성대사질환학회지
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• 제5권1호
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• pp.9-17
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• 2005

Homocystinuria is a metabolic disorder caused by a deficiency of cystathionine ${\beta}$-synthase (CBS). Patients with homocystinuria show clinical symptoms such as mental retardation, lens dislocation, vascular disease with life-threatening thromboembolisms and skeletal deformities. Generally, the major treatments for CBS deficiency include pharmacologic doses of pyridoxine or dietary restriction of methionine. However, there is no effective treatment for this disease up till today and gene therapy can be an attractive novel approach to treatment of the disease. We investigated whether a recombinant adeno-associated virus could be used as a CBS gene transfer vector to reduce the excessive homocysteine level in the homocystinuria mouse model. Recombinant adeno-associated virus vector encoding the human CBS gene (rAAV-hCBS), driven by EF1-a promoter, was infused into CBS-deficient mice ($CBS^{-/-}$) via intramuscular (IM) and intraperitoneal (IP) injection. IP injection was more efficient than IM injection for prolongation of lives and reduction of plasma homocysteine levels. After 2 weeks of gene transfer by IP injection, serum homocysteine level was significantly decreased in treated mice compared with the age-matched controls and the life span was extended about 1.5 times. Also, increased expression of CBS gene was observed by immunohistochemical staining in livers of treated $CBS^{-/-}$ mice and microvesicular lipid droplets was decreased in cytoplasm of liver. These results demonstrate the possibility and efficacy of gene therapy by AAV gene transfer in homocystinuria mice.