• The Journal of Internal Korean Medicine
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• v.45 no.1
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• pp.11-30
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• 2024

Objectives: This study evaluated the effects of Ssanghwa-tang (SHT) on lung injury and muscle loss in a COPD mouse model. Methods: C57BL/6 mice were challenged with cigarette smoke extract and lipopolysaccharide, and then treated with two concentrations of SHT (250 and 500 mg/kg). After sacrifice, the bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed by cytospin, ELISA, real-time PCR, flow cytometry analysis, and H&E and Masson's trichrome staining. The grip strength of COPD mice was measured using a grip strength meter. The running time of COPD mice was measured by a treadmill test. Muscle tissue of the quadriceps was stained with H&E and Masson's trichrome staining. Results: SHT significantly inhibited the increase in neutrophil numbers in BALF and significantly decreased immune cell activity in BALF and lung tissue. It also significantly inhibited the increase in TNF-α, IL-17, and MIP2 in BALF. Real-time PCR analysis revealed that the mRNA expression of TNF-α, IL-17, MIP2, and TRPV1 in lung tissue showed a significant decrease compared with the control group. Lung tissue damage was significantly reduced in the histological analysis. The grip strength and running time of the COPD mice showed a significant decrease compared with the control group. In histological staining, SHT was found to reduce the damage to muscle tissue. Conclusions: This study indicates that SHT can be used as a therapeutic agent for COPD patients by inhibiting lung injury and muscle loss.

• Korean Journal of Acupuncture
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• v.41 no.2
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• pp.33-42
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• 2024

Objectives : The sigma-1 receptor is implicated in stress, depression, psychostimulant sensitization, and addiction vulnerability. Prior studies have indicated that ethanol exposure modulates sigma-1 receptor activity within the Ventral Tegmental Area (VTA). Here, we explore the sub-mechanisms underlying sigma-1 receptor activity induced by HT7 (Shinmun) stimulation in behavioral alterations following acute ethanol (ETOH) administration. Methods : Male Wistar rats were investigated for pro- and anti-inflammatory markers after injection of ETOH (1 g/kg) using cytokine enzyme-linked immunosorbent assay (ELISA)s. After confirming that HT7 stimulation changed the total distance traveled in the open field test (OFT), protein changes in the Ventral tegmental area (VTA) were measured by Western blotting. The expression level of inducible nitric oxide synthase (iNOS) after administration of a sigma-1 receptor antagonist (dihydrobromide 1047; BD1047, 10 mg/kg i.p.) and Shenmen (HT7) stimulation was compared. Results : As a result, acute ETOH administration increased proinflammatory marker levels (TNF-𝛼 and IL-6). HT7 stimulation restored the total distance response after acute ethanol administration. In addition, in the VTA, the levels of a microglial marker (iNOS), sigma-1 receptor and protein kinase C, which are predicted to be involved in up- and downregulation, were restored by HT7 stimulation. In particular, HT7 stimulation modulates iNOS expression through effects similar to BD treatment. This study suggests that the stimulatory effect of HT7 may be driven by microglial activation. Conclusions : Microglial activity is regulated by sigma-1 receptor, and sigma-1 receptor activity is regulated by HT7 stimulation. Significantly, we demonstrate that HT7 stimulation ameliorates behavioral alterations induced by acute ETOH administration through microglial activation within the VTA.

• IMMUNE NETWORK
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• v.22 no.3
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• pp.24.1-24.12
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• 2022

Coronavirus disease 2019 (COVID-19) vaccination in immunocompromised, especially transplant recipients, may induce a weaker immune response. But there are limited data on the immune response after COVID-19 vaccination in liver transplant (LT) recipients, especially on the comparison of Ab responses after different vaccine platforms between mRNA and adenoviral vector vaccines. Thus, we conducted a prospective study on LT recipients who received two doses of the ChAdOx1 nCoV-19 (ChAdOx1), mRNA-1273, or BNT162b2 vaccines compared with healthy healthcare workers (HCWs). SARS-CoV-2 S1-specific IgG Ab titers were measured using ELISA. Overall, 89 LT recipients (ChAdOx1, n=16 [18%]) or mRNA vaccines (mRNA-1273 vaccine, n=23 [26%]; BNT162b2 vaccine, n=50 [56%]) received 3 different vaccines. Of them, 16 (18%) had a positive Ab response after one dose of COVID-19 vaccine and 62 (73%) after 2 doses. However, the median Ab titer after two doses of mRNA vaccines was significantly higher (44.6 IU/ml) than after two doses of ChAdOx1 (19.2 IU/ml, p=0.04). The longer time interval from transplantation was significantly associated with high Ab titers after two doses of vaccine (p=0.003). However, mycophenolic acid use was not associated with Ab titers (p=0.53). In conclusion, about 3-quarters of LT recipients had a positive Ab response after 2 doses of vaccine, and the mRNA vaccines induced higher Ab responses than the ChAdOx1 vaccine.

• IMMUNE NETWORK
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• v.20 no.5
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• pp.40.1-40.15
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• 2020

The protein encoded by the Gene Associated with Retinoid-Interferon-Induced Mortality-19 (GRIM-19) is located in the mitochondrial inner membrane and is homologous to the NADH dehydrogenase 1-alpha subcomplex subunit 13 of the electron transport chain. Multiple sclerosis (MS) is a demyelinating disease that damages the brain and spinal cord. Although both the cause and mechanism of MS progression remain unclear, it is accepted that an immune disorder is involved. We explored whether GRIM-19 ameliorated MS by increasing the levels of inflammatory cytokines and immune cells; we used a mouse model of experimental autoimmune encephalomyelitis (EAE) to this end. Six-to-eight-week-old male C57BL/6, IFNγ-knockout (KO), and GRIM-19 transgenic mice were used; EAE was induced in all strains. A GRIM-19 overexpression vector (GRIM19 OVN) was electrophoretically injected intravenously. The levels of Th1 and Th17 cells were measured via flow cytometry, immunofluorescence, and immunohistochemical analysis. IL-17A and IFNγ expression levels were assessed via ELISA and quantitative PCR. IL-17A expression decreased and IFNγ expression increased in EAE mice that received injections of the GRIM-19 OVN. GRIM19 transgenic mice expressed more IFNγ than did wild-type mice; this inhibited EAE development. However, the effect of GRIM-19 overexpression on the EAE of IFNγ-KO mice did not differ from that of the empty vector. GRIM-19 expression was therapeutic for EAE mice, elevating the IFNγ level. GRIM-19 regulated the Th17/Treg cell balance.

• Korean Journal of Veterinary Research
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• v.64 no.2
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• pp.17.1-17.10
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• 2024

This study examined the colostrum nutritive composition, immunoglobulin (Ig), and microbial community in Holstein and Jersey dairy cows according to the time after calving. The experiment used seven Holstein and three Jersey dairy cows. Colostrum was collected immediately after calf calving, 12, and 24 hours, and stored at -80℃ until analysis. An analysis of the nutritive composition in colostrum was performed using LactoScop. The immune indicators were analyzed using an ELISA Kit, and the microbial community was assessed using a Macrogen Inc. The protein level was high in all colostrum samples from Holstein dairy cows compared with Jersey dairy cows, but there was no significant difference according to the time after calving. Immune index analysis revealed high IgG and IgA concentrations in the colostrum of Holstein cows immediately after calving and 12 and 24 hours after calving, but the differences were not significant. The microbial community at the genus level revealed Staphylococcus to be predominant at a high rate in the colostrum of Holstein dairy cows and Enterococcus in Jersey dairy cows 12 hours after calving. Pseudomonas was predominant at a high rate in the colostrum of Jersey lactating cows immediately and 12 hours after calving. Chryseobacterium was predominant at a high rate in Holstein dairy cows 12 and 24 hours after calving. In conclusion, these results are expected to be used as research data on the correlation between quality, immunity, and microbial community in the colostrum.

• Journal of Microbiology and Biotechnology
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• v.34 no.4
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• pp.920-929
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• 2024

As a pivotal defensive line against multitudinous malignant tumors, natural killer (NK) cells exist in the tumor microenvironment (TME). RAD18 E3 Ubiquitin Protein Ligase (RAD18) has been reported to foster the malignant progression of multiple cancers, but its effect on NK function has not been mined. Here, the study was designed to mine the mechanism by which RAD18 regulates the killing effect of NK cells on colorectal cancer (CRC) cells. Expression of E2F Transcription Factor 7 (E2F7) and RAD18 in CRC tissues, their correlation, binding sites, and RAD18 enrichment pathway were analyzed by bioinformatics. Expression of E2F7 and RAD18 in cells was assayed by qRT-PCR and western blot. Dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay verified the regulatory relationship between E2F7 and RAD18. CCK-8 assay was utilized to assay cell viability, colony formation assay to detect cell proliferation, lactate dehydrogenase (LDH) test to assay NK cell cytotoxicity, ELISA to assay levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), and immunofluorescence to detect expression of toxic molecules perforin and granzyme B. High expression of RAD18 and E2F7 was found in CRC tissues and cells. Silencing RAD18 could hamper the proliferation of CRC cells, foster viability and cytotoxicity of NK cells, and increase the secretion of GM-CSF, TNF-α, IFN-γ as well as the expression of perforin and granzyme B. Additionally, ChIP and dual-luciferase reporter assay ascertained the binding relationship between RAD18 promoter region and E2F7. E2F7 could activate the transcription of RAD18, and silencing RAD18 reversed the inhibitory effect of E2F7 overexpression on NK cell killing. This work clarified the inhibitory effect of the E2F7/RAD18 axis on NK cell killing in CRC, and proffered a new direction for immunotherapy of CRC in targeted immune microenvironment.

• International Journal of Stem Cells
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• v.15 no.4
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• pp.395-404
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• 2022

Background and Objectives: Chronic obstructive pulmonary disease (COPD) is a common, frequently-occurring disease and poses a major health concern. Unfortunately, there is current no effective treatment for COPD, particularly emphysema. Recently, experimental treatment of COPD using mesenchymal stem cells (MSCs) mainly focused on bone marrow-derived MSCs (BM-MSCs). Human umbilical cord-derived MSCs (hUC-MSCs) have more advantages compared to BM-MSCs. However, studies on the role of hUC-MSCs in management of COPD are limited. This study sought to explore the role of hUC-MSCs and its action mechanisms in a rat model of VEGF receptor blocker SU5416-injured emphysema. Methods and Results: hUC-MSCs were characterized by immunophenotype and differentiation analysis. Rats were divided into four groups: Control, Control+MSC, SU5416 and SU5416+MSC. Rats in model group were administered with SU5416 for three weeks. At the end of the second week after SU5416 administration, model group were infused with 3×10⁶ hUC-MSCs through tail vein. After 14 days from hUC-MSCs transplantation, rats were euthanized and data were analyzed. HE staining and mean linear intercepts showed that SU5416-treated rats exhibited typical emphysema while emphysematous changes in model rats after hUC-MSCs transplantation disappeared completely and were restored to normal phenotype. Furthermore, hUC-MSCs inhibited apoptosis as shown by TUNEL and Western blotting. ELISA and Western blotting showed hUC-MSCs rescued VEGF-VEGFR2-AKT pathway in emphysematous lungs. Conclusions: The findings show that hUC-MSCs effectively repair the emphysema injury. This study provides the first evidence that hUC-MSCs inhibit apoptosis via rescuing VEGF- VEGFR2-AKT pathway in a rat model of emphysema.

• Herbal Formula Science
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• v.32 no.3
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• pp.297-310
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• 2024

Objective : This study was conducted to investigate the anti-inflammatory and laxative effects of Polycan in TNF-α-treated HT-29 intestinal epithelial cells and loperamide-induced constipation in vivo models, respectively. Methods : To evaluate the anti-inflammatory effects of Polycan, HT-29 cells were treated with TNF-α in the presence or absence of Polycan. IL-8 production was measured by enzyme-linked immunosorbent assay (ELISA). MAPK phosphorylation, nuclear translocation of NF-κB, and phosphorylation of IκB were assessed by Western blot analysis. To investigate the laxative effects of Polycan, 6-week-old SD rats (8 female rats per group) were orally administered Polycan or Chicory Fiber as a positive control for 4 weeks, and constipation was induced with loperamide treatment for 10 days before sacrifice. One day before sacrifice, a charcoal meal was administered to evaluate intestinal transit times. The periodically collected feces were used to assess the number of fecal pellets and fecal water content. Results : Polycan inhibited TNF-α-induced IL-8 expression in dose-dependent manner. Furthermore, Polycan suppressed TNF-α-induced phosphorylation of MAPKs (ERK1/2, p38 and JNK), degradation of Iκ-Bα and nuclear translocation of NF-κB. In an in vivo constipation model, the number of fecal pellets per food intake was significantly increased in rats administered with Polycan, both 1 day and 7 days after loperamide treatment. The water content of fecal pellets was restored in the Polycan groups starting 7 days after loperamide treatment. In addition, Polycan intake significantly enhanced the gastrointestinal transit ratio of a charcoal meal but reduced the number of intestinal fecal pellets. Conclusions : These results suggest that Polycan suppressed TNF-α-induced inflammation by blocking both the MAPK and NF-κB pathways in HT-29 cells. Additionally, in a loperamide-induced constipation model, Polycan showed clear laxative effects by increasing the number of fecal pellets, fecal water content, and intestinal transit ratio of a charcoal meal.

• Journal of Ginseng Research
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• v.48 no.5
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• pp.494-503
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• 2024

Background: With the prevalence of dietary supplements, the use of combinations of herbs and drugs is gradually increasing, together with the risk of drug interactions. In our clinical work, we unexpectedly found that the combination of Panax notoginseng and warfarin, which are herbs that activate blood circulation and remove blood stasis, showed antagonistic effects instead. The purpose of this study was to evaluate the drug interaction between Panax notoginseng saponins (PNS) and warfarin, the main active ingredient of Panax notoginseng, and to explore the interaction mechanism. Methods: The effects and mechanisms of PNS on the pharmacodynamics and pharmacokinetics of warfarin were explored mainly in Sprague-Dawley rats and HepG2 cells. Elisa was used to detect the concentrations of coagulation factors, HPLC-MS to detect the blood concentrations of warfarin in rats, immunoblotting was employed to examine protein levels, qRT-PCR to detect mRNA levels, cellular immunofluorescence to detect the localization of NR1I3, and dual luciferase to verify the binding of miR-214-3p and NR1I3. Results: PNS significantly accelerated warfarin metabolism and reduced its efficacy, accompanied by increased expression of NR1I3 and CYP2C9. Interference with NR1I3 rescued the accelerated metabolism of warfarin induce by PNS co-administration. In addition, we demonstrated that PNS significantly reduced miR-214-3p expression, whereas miR-214-3p overexpression reduced NR1I3 and CYP2C9 expression, resulting in a weakened antagonistic effect of PNS on warfarin. Additionally, we found that miR-214-3p bound directly to NR1I3 3'-UTR and significantly downregulated NR1I3 expression. Conclusion: Our study demonstrated that PNS accelerates warfarin metabolism and reduces its pharmacodynamics by downregulating miR-214-3p, leading to increased expression of its target gene NR1I3, these findings provide new insights for clinical drug applications to avoid adverse effects.

• Herbal Formula Science
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• v.32 no.2
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• pp.155-179
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• 2024

Objective : Danggwisu-san (DGSS) is an herbal formula that has been mainly used in the East Asia for the treatment of bruise, sprain and external injury. The cause of this pain is that Qi and blood become tangled and do not circulate well. DGSS can improve the tangled situation and make it well-circulated. The present study evaluated the anti-inflammatory effects of DGSS on Raw 264.7 cells and in rats with paw edema. Methods : Cell viability was measured using the MTT assay. The amount of nitric oxide (NO) production was measured the amount of nitrite content in the cultured medium using Griess reagent. The amount of tumor necrosis factor-α, monocyte chemoattractant protein 1, interleukin (IL)-1βand IL-6 in the cultured supernatant were measured by ELISA kit. Proteins expression were detected by Western blot. Furthermore, the effect of DGSS on acute inflammation was observed in rat paw edema model. Results : The DGSS ameliorates the lipopolysaccharide-activated changes in NO production, iNOS expression and pro-inflammatory cytokines. Additionally, DGSS significantly suppressed expression of p-JNK, p-ERK and nuclear NF-κB. As expected, in rat paw edema study, 1.0 g/kg of DGSS significantly reduced the carrageenan-induced paw edema and iNOS expression for 1-4 h. Moreover, administration of 1.0 g/kg (4 days) of DGSS used in this study did not show any significant change on ALT and AST. Conclusion : These results demonstrate that DGSS has anti-inflammatory effects in vitro and in vivo. Therefore, this present study can put scientific evidences up for the anti-inflammatory effect of DGSS.