• The Journal of Internal Korean Medicine
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• v.21 no.2
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• pp.259-266
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• 2000

Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.11
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• pp.1837-1847
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• 2020

Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.

• IMMUNE NETWORK
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• v.6 no.3
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• pp.154-162
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• 2006

Background: Dendritic cell (DC)-based cancer immunotherapy is studied for several years. However, it is mainly derived from autologous PBMC or leukapheresis from patient, which has limitations about yield and ability of DC production according to individual status. In order to solve these problems, inquiries about allogeneic DCs are performed but there are no preclinical trial answers for effect or toxicity of allogeneic DC to use for clinical trial. In this study, we compared the anti-tumor effect of allogeneic and autologous DCs from mouse bone marrow stem cells in mouse metastatic melanoma model. Methods: B16F10 melanoma cells ($5{\times}10^4$/mouse) were injected intravenously into the C57BL/6 mouse. Therapeutic DCs were differentiated from autologous (C57BL/6: CDC) or allogeneic (B6C3F1: BDC) bone marrow stem cells with GM-CSF, SCF and IL-4 for 13days and pulsed with B16F10 tumor cell lysate (Blys) for 18hrs. DC intra-peritoneal injections began on the 8th day after the tumor cell injection by twice with one week interval. Results: Anti-tumor response was observed by DC treatment without any toxicity especially in allogeneic DC treated mice (tumor burden score: $2.667{\pm}0.184,\;2.500{\pm}0.463,\;2.000{\pm}0.286,\;1.500{\pm}0.286,\;1.667 {\pm}0.297$ for saline, CDC/unpulsed-DC: U-DC, CDC/Blys-DC, BDC/U-DC and BDC/Blys-DC, respectively). IFN-${\gamma}$ secretion was significantly increased in allogeneic DC group stimulated with B16F10 cell lysate ($2,643.3{\pm}5,89.7,\;8,561.5{\pm}2,204.9.\;6,901.2{\pm}141.1pg/1{\times}10^6$ cells for saline, BDC/U-DC and BDC/Blys-DC, respectively) with increased NK cell activity. Conclusion: Conclusively, promising data was obtained that allogeneic DC can be used for DC-based cancer immunotherapy.

• Herbal Formula Science
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• v.21 no.1
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• pp.51-70
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• 2013

Objectives : The object of this study was to observe anticancer and related immunomodulatory effects of Insamyangyoung-tang extracts (ISYYTe) on non-small cell lung carcinoma (squamous epithelial carcinoma), NCI-H520, xenograft Balb/c nu-nu nude mice. Methods : Three different dosages of ISYYTe, 50, 100 and 200 mg/kg were orally administered once a day for 42 days from 11 days after tumor cell inoculation. Six groups, which are intact control, tumor bearing control, 5-fluorouracil (FU) 30 mg/kg, ISYYTe 50 mg/kg, ISYYTe 100 mg/kg, ISYYTe 200 mg/kg, each of 8 mice per group were used in the present study. Changes on the body weight, tumor volume and weight, lymphatic organ (spleen and popliteal lymph node), serum interferon (IFN)-${\gamma}$ levels, splenocytes NK cell activity and peritoneal macrophage activities, splenic tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-10 contents were observed with tumor mass and lymphatic organ histopathology to detect anticancer and immunomodulatory effects. Results : As results of ISYYTe 50, 100 and 200 mg/kg treatment, decreases in the tumor volumes and weights were detected. At histopathological observations, decreases of tumor cell volumes in tumor masses were dose-dependently decreased mediated by increases of apoptosis among tumor cells by treatment of all three different dosages of ISYYTe. As results of tumor cell inoculation, marked decreases of spleen and popliteal lymph node weights, serum IFN-${\gamma}$, splenic TNF-${\alpha}$, IL-$1{\beta}$ and IL-10 contents and splenocytes were observed with histopathological atrophic changes of spleen and popliteal lymph nodes. Conclusions : Over 50 mg/kg of ISYYTe showed favorable anticancer effects on the NCI-H520 cell xenograft with immunomodulatory effects. Although relatively lower anticancer effects were observed in ISYYTe 200 mg/kg treated mice as compared with 5-FU 30 mg/kg treated mice, there are no meaningful favorable immunomodulatory effects were observed after 5-FU treatment in the present study.

• Reproductive and Developmental Biology
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• v.32 no.1
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• pp.1-7
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• 2008

The SCID-repopulation cells(SRCs) assay has been widely used to determine the self-renewal capacity of hematopoietic stem cells (HSCs). In this study, we tested the repopulating efficiency of porcine bone marrow derived hematopoietic stem cells using nonobese diabetic/severe combined immunodieficient (NOD/SCID) mice which was inherited immunodeficiency mire with defect of T cells, B cells, and low activity of NK cells. We transplanted porcine bone marrow hematopoietic stem/progenitor cells with intraperitoneal injection into neonate NOD/SCID mice. We confirmed efficient reconstitution activity of inoculated porcine hematopoietis cells in variety of organs of NOD/SCID mice. Interestingly, pig $CD3^+$ T lymphocytes detected with high level in liver($15.6{\pm}3.7%$), spleen($5.6{\pm}3.0%$), thymus($1.5{\pm}1.3%$), and BM($2.3{\pm}0.9%$), respectively. These data imply that microenvironment of neonate NOD/SCID mice is very efficient for proliferation and differentiation of porcine T cells, and can be useful for the study of T cells development and renogeneic organ transplantation.

• Journal of Ginseng Research
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• v.30 no.1
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• pp.41-48
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• 2006

In the browning reaction of Korean ginseng, it appears that enzymatic and non-enzymatic browning reaction occurred In initial stage of steaming fresh ginseng at low temperature, and then non-enzymatic browning reaction followed in the drying period after steaming. Browning reaction of red ginseng occurred between $60{\sim}90$ min of steaming at $100^{\circ}C$, and browning pigments of red ginseng were mostly water soluble substances. The structural characteristics of water soluble browning reaction products(WS-BRPs) isolated from Korean red ginseng were showed the presence of hydroxyl, amide carbonyl and aliphatic methane groups. From sugar analysis it was identified that L and S-1, melanoidins isolated from red ginseng, contained two kinds of sugars, glucose and xylose, and the other melanoidin S-2 contained the previous and fructose. In order to find out pertinent methods for the acceleration of browning during ginseng processing, various treatment were made on fresh ginseng with sugars, amino acids and inorganic nitrogenous compounds and the extent of browning was measured. Among sugar tested, maltose resulted in the greatest acceleration of browning followed in decreasing order by glucose and lactose, whereas pentoses, fructose, sucrose and raffinose had negligible effect. A marked browning occurred in ginseng treated with basic amino acids, while the extent of browning was not greatly increased when ginseng was treated with aliphatic amino acids, hydroxyl amino acids, or acidic amino acids. The brown color intensity gradually increased with an increase of glucose concentration far up to 0.5M. L, S-1, and S-2 were found to have an ability to donate hydrogen to DPPH, and also they had anti-oxidative activity in the experiments of hydrogen peroxide scavenging, inhibitory activity in the formation of MDA from linoleic acid, auto oxidation of ok-brain homogenates, lipid peroxidation by the enzymatic and non-enzymatic system in liver microsome fraction, and mitochondrial fraction etc. The amounts of acidic polysaccharide(AP) in red ginseng were higher than those of wild and cultured Panax quinquefolius, Panax notoginseng as well as white ginseng (Panax ginseng). In white ginseng, the AP amount is no difference in root ages or sizes, also, the AP amount of ginseng body was similar to that of rhizome, but was higher than that of leaf and epidermis. Addition of red ginseng acidic polysaccharide(RGAP) increased production of nitric oxide(NO) and tumor necrosis factor (TNF)-$\alpha$ in the rodent macrophage cultures, and treatment of RGAP in vivo stimulated tumoricidal activities of natural killer (NK) cells.

• Journal of Korean Academy of Fundamentals of Nursing
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• v.7 no.1
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• pp.109-121
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• 2000

Increasingly nursing science is embracing the concepts and methodology derived from psycho-neuroimmunology. It has been previously shown that stress increases and immune function declines in students undergoing examinations. To date, however, no many studies have been reported on stress levels, immune function and interventions in Korean students undergoing their first clinical nursing rotation. It was proposed that nursing students during their first clinical rotation experience increase in stress because of the novelty of the situation and their lack of clinical knowledge. It was also hypothesized that biofeedback and progressive relaxation, methods of self-regulation of involuntary autonomic nervous system responses, would reduce the stress response. The purpose of this study is to test the effectiveness of progressive muscle laxation using biofeedback The effectiveness of the experimental methods was tested by measuring the degree of symptoms of stress (SOS) and the values of ephinephrine, pulse rate, blood pressure and natural killer cells. The subjects of this study were thirty nursing students divided into two groups: experimental group was progressive muscle relaxation group using biofeedback and control group. This study was conducted for 8 weeks of clinical practice. Biofeedback training was done by software developed by J&J company (1-410 form for progressive muscle training). Progressive muscle relaxation training according to Jacobson's Theory was done by messaged word from biofeedback. The data was analyzed using Chronbach' ${\alpha}$ and t-test of the SPSS program and the significance level of statistics was 5%. The results of the study were : 1) The progressive muscle relaxation training using biofeedback was effective for the reduction of symptoms of stress(t=-4.248, p<.001) under clinical practice stress conditions. 2) The progressive muscle relaxation training using biofeedback was not effective for the values of epinephrine(t=-1.294, p=.206). 3) The progressive muscle relaxation training using biofeedback was effective for the reduction of systolic blood pressure (t=-2.757, p=.01). 4) The progressive muscle relaxation training using biofeedback was effective for the reduction of diastolic blood pressure (p=-2.032, 0=.05). 5) The progressive muscle relaxation training using biofeedback was not effective for the reduction of pulse rate(t=-15, p=.988). 6) The progressive muscle relaxation training using biofeedback was effective for the maintenance of natural killer cells (t=2.381, p=02). The first clinical rotation for student nurses is a stressful experience as seen by the rise in the SOS in the control group. Biofeedback using progressive muscle relaxation were effective in preventing the rise of symptoms of stress and the blood pressure means when comparing the pre to post clinical experience, The mean natural killer cell count was depressed in the control group but not significantly different in the experimental groups, It is proposed here that stress via the hypothalamic - pituitary - adrenal axis suppressed the NK cell count whereas the relaxation methods prevented the rise in stress and the resulting immune depression. We recommend relaxation techniques using biofeedback as a health promotion technique to reduce psychological stress. In summary. the progressive muscle relaxation training using biofeedback was effective for the reduction of symptoms of stress under clinical practice stress conditions.

• Korean Journal of Korean Medical Institute of Dermatology and Aesthetics
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• v.1 no.1
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• pp.53-90
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• 2005

Purpose: Contents of astragaloside I, II and IV, cytotoxicity, anticancer activity, immunomodulatory activity and antioxidant capacity were to be compared as a function of the cultivated years as one, three, five and seven years. Method: Major components of Astragali membranacei Radix were separated as astragaloside I, astragaloside II, astragaloside IV by HPLC analysis. Cytotoxicity and anticancer activities were measured by MTT and SRB assay. For immunomodulatory activity, the secretion of IL -6 and $TNF-{\alpha}$, NK cell activation and macrophage activation were observed as well as kinetics of responding to human T cells by a microphysiometer. In vitro antioxidant activities were measured by several radical scavenging activities of superoxide anion radican, DPPH, LDL and linoleic acid. For in vivo activity, the activation of SOD, GSH-px, catalase, ALDH and ADH was measured as well the relative weight of liver. Result : 1. For HPLC analysis, the contents of all of astragaloside I, astragaloside II, astragaloside IV were in order of three, five, one and seven years. 2. The cytotoxicity of normal human lung cell line, HEL299 showed lower than 18% in adding 0.25 mg/ml, and 28.9% in adding 1.0 mg/ml of water extract of seven year root. For methanol extracts, three year root showed highest cytotoxicity as 35.2 % and there was no difference between the cultivated years. 3. For anticancer activities, methanol extracts of one and three year roots showed relatively high inhibition of human stomach cancer cells, AGS, breast cancer cells, MCF-7, lung cancer cells, A549 and liver cancer cell, Hep3B as well as high selectivities. 4. The water extract of seven year root could yield high secretion of IL-6 from both human Band T cells while the methanol extracts of three and five year roots secreted high amounts of IL-6 and $TNF-{\alpha}$ from both Band T cells. 5. As a result of in vitro antioxidant activities, both water and methanol extracts from five and seven year roots showed high activities for superoxide anion radical scavenging activity, inhibiting linoleic acid peroxide and contents of total phenols. 6. For in vivo tests, Mn-SOD and GSH-px activities and weight of liver were better in adding seven year root. For ALDH activity one year root was better and for ADH activity five year root. Overall speaking, seven year root showed relatively better antioxidant activities. Conclusion:There was difference of the contents of astragaloside I, astragaloside II, astragaloside IV according to cultivation year. Methanol extract showed better activities of anticancer and immune activation rather than water extract Interestingly enough, for methanol extracts, overall activities were improved as the cultivation year increased. There might be further investigation required for the clinical uses of the results as several biological activities varied according to the cultivated year of Astragali membranacei Radix.

• Safety and Health at Work
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• v.13 no.2
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• pp.248-254
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• 2022

Background: Occupational hazards in crop farms vary diversely based on different field operations as soil management, harvesting processes, pesticide, or fertilizer application. We aimed at evaluating the immunological status of crop farmers, as limited systematic investigations on immune alteration involved with crop farming have been reported yet. Methods: Immunological parameters including plasma immunoglobulin level, major peripheral immune cells distribution, and level of cytokine production from activated T cell were conducted. Nineteen grape orchard, 48 onion open-field, and 21 rose greenhouse farmers were participated. Results: Significantly low proportion of natural killer (NK) cell, a core cell for innate immunity, was revealed in the grape farmers (19.8±3.3%) in comparison to the onion farmers (26.4±3.1%) and the rose farmers (26.9±2.5%), whereas cytotoxic T lymphocyte proportion was lower in the grape and the onion farmers than the rose farmers. The proportion of NKT cell, an immune cell implicated with allergic response, was significantly higher in the grape (2.3±0.3%) and the onion (1.6±0.8%) farmers compared with the rose farmers (1.0±0.4%). A significantly decreased interferon-gamma:interleukin-13 ratio was observed from ex vivo stimulated peripheral blood mononuclear cells of grape farmers compared with the other two groups. The grape farmers revealed the lowest levels of plasma IgG1 and IgG4, and their plasma IgE level was not significantly different from that of the onion or the rose farmers. Conclusion: Our finding suggests the high vulnerability of workplace-mediated allergic immunity in grape orchard farmers followed by open-field onion farmers and then the rose greenhouse farmers.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.350-354
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• 2010

Laminaria japonica polysaccharides (LP) were prepared from L. japonica through hot water extraction, ultrafiltration and gel chromatography. In this study, we investigated the immunomodulating activity of LP (0.25-1 mg/mL) on the mitogen/alloantigen reactive proliferation and killing activity of the Balb/c mouse splenocytes. The LP directly induced the proliferation of splenocytes that was stimulated with mitogen or alloantigen in a dose-dependent manner. The killing activity of cytotoxic T lymphocytes (CTLs) and lymphokine activated killer cells (LAKs) were enhanced significantly in the LP treated cells. Also, the treatment of splenocytes with LP increased production of interleukin-2 (IL-2). These results suggest that polysaccharides from L. japonica show a substantial immunomodulating activity in mouse immune cells.