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Increase of Plasma IL-12/p40 Ratio Induced by the Combined Therapy of DNA Vaccine and Lamivudine Correlates with Sustained Viremia Control in CHB Carriers

  • Im, Se-Jin;Yang, Se-Hwan;Yoon, Seung-Kew;Sung, Young-Chul
    • IMMUNE NETWORK
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    • v.9 no.1
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    • pp.20-26
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    • 2009
  • We previously reported that $IFN-{\gamma}$ producing T cell responses induced by the combined therapy of DNA vaccine and lamivudine for one year are important for the induction of sustained virological response (SVR). However, $IFN-{\gamma}$ production is not sufficient to predict sustained viremia control in chronic hepatitis B (CHB) carriers treated. Methods: Twelve CHB carriers were intramuscularly immunized 12 times at a 4-week interval with 8mg of HBV DNA vaccine during the standard lamivudine treatment (100mg/daily/1 year). The level of cytokines during and after the combined therapy in plasma of all 12 CHB carriers treated was determined by each ELISA kit. Six out of 12 CHB carriers revisited the clinic, and their HBV DNA levels were examined. Results: The combined therapy increased plasma IL-12 and IL-12/p40 ratio during the treatment (baseline vs. peak level: $41.8{\pm}8.3$ vs. $163.1{\pm}29.2\;pg/ml$; p<0.01 and $0.96{\pm}0.25$ vs. $3.58{\pm}0.86$; p<0.01, respectively), and the peak level of plasma IL-12 and IL-12/p40 ratio was evoked at 6 to 10 months during the combined therapy. In particular, CHB carriers with SVR had two and three-fold higher level of the peak plasma IL-12 and plasma IL-12/p40 ratio than non-virological responders (NVRs), respectively ($218.0{\pm}41.4$ vs. $108.1{\pm}28.6\;pg/ml$; p=0.09 and $5.35{\pm}1.38$ vs. $1.80{\pm}0.29$; p<0.05, respectively), while p40 level was consistent during the combined therapy. In addition, there was no significant temporal correlation between the peak IL-12/p40 ratio and the elevation of serum alanine amino-transferase (ALT) in this study, contrast to $IFN-{\alpha}$ therapy which induced peak IL-12 level following ALT flares. Conclusion: Our results indicate that the combined therapy induces the increase of plasma IL-12 and IL-12/p40 ratio, which are associated with long-term SVR in CHB carriers.

Antitumor and immunomodulatory effects of a hot water extract of Phellinus baumii using Sarcoma-180 in the mouse (Pheflinus baumii 자실체 추출물의 Sarcoma-180 에 대한 항암 및 면역효과)

  • Ha, Hyo-Cheol;Kim, Hyun-Pyo;Sim, Ji Young;Chang, Yun-Hee;Kim, Hyun Su
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.169-174
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    • 2004
  • A hot water extract(HWE-P4) was separated from the fruit bodies of PMO-P4, and its antitumor and immunomodulatory activities against sarcoma-180 in ICR mice were investigated. The internal transcribed spacer(ITS) regions from PMO-P4 were amplified using polymerase chain reaction(PCR) and sequenced. The results revealed that PMO-P4 was belong to the Phellinus baumii. When oral administration at the dose of 160mg/kg/day in the mice until the end of the experiment with 2 week's pre-feeding of the HWE-P4, the survival rate of the mice was 152% for 50days after the inoculation of sarcoma-180 and the suppression rate of the tumor growth was 35.3%(p<0.05) for 28 days after inoculation of sarcoma-180. The HWE-P4 increased 71.4% of the CD4/CD8 ratio and 5-fold of the expression of CD25(IL-2 receptor chain) compared with the control. From these results, the antitumor activity of HWE-P4 is exerted through its immunomodulating activity on the host's immune system.

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The Palliative Care and Hospice for the People Living with HIV (HIV 감염인을 위한 완화의료와 호스피스)

  • Choi, Jae-Phil
    • Journal of Hospice and Palliative Care
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    • v.20 no.3
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    • pp.159-166
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    • 2017
  • According to the advance of antiretroviral regimen and the early treatment strategy, people living with human immunodeficiency virus (PLWH) are achieving the goal of virologic suppression and immune restoration. Most of them no more die of acquired immunodeficiency syndrome (AIDS) defining illnesses, and become older with chronic comorbidities such as cardiovascular, metabolic, hepatic, renal and neurological diseases. However some PLWH still visit hospitals as late presenters with very low CD4+ T cell counts, so that they suffer AIDS defining illnesses to die or experience severe neurological complications resulting in disabilities. Early palliative interventions are needed on the various symptoms of PLWH. Thus far chronic pains such as distal symmetric sensory polyneuropathies have been underevaluated. Active pain-relieving interventions are important to them. Recently we define end of life condition of human immunodeficiency virus (HIV) or eligibility to hospice care after adjusting current status of HIV treatment. Hospice teams should pay attention to the specific medical conditions, psychological needs, and social circumstances of PLWH. With just standard precautions as common infection control measures, general hospice cares can be provided to them like to other hospices subjects. For giving PLWH opportunities to have the end of life with value and dignity, hospice multidisciplinary team should intervene them early and aggressively. Now we need more clinical experiences and institutional improvements.

A Study of the Bystander Effect and Its Enhancement in HSV-TK Gene Therapy Using a Murine Neuroblastoma Model (마우스 신경모세포종 모델을 이용한 HSV-TK 유전자 치료에서 Bystander 효과 및 증폭에 관한 연구)

  • Cho, Hyun Sang;Kim, Moon Kyu;Park, Chong Young
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.354-361
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    • 2002
  • Purpose : Selective introduction of genes conferring chemosensitivity into proliferating tumor cells may be used to treat cancer. We first investigated the bystander effect of retrovirus-mediated gene transfer of herpes simplex virus thymidine kinase(HSV-TK) gene to murine neuroblstoma cell line(neuro-2a) in vitro and in vivo. Second, we examined the mechanism and its enhancement of the bystander effect in murine neuroblastoma. Methods : To investigate the bystander effect, we studied tumor growth and survival time after HSV-TK/ganciclovir(GCV) treatment in a syngenic A/J mouse neuroblastoma model by mixing various ratios of HSV-TK-expressing neuro-2a cells with wild type neuro-2a cells followed by GCV treatment. To investigate the mechanism of the bystander effect in murine neuroblastoma, immunohistochemistry using connexin 43, CD4 and CD8-specific monoclonal antibodies was analyzed. We studied whether IL-2-secreting neuro-2a cells(neuro-2a/IL-2) would potentiate the bystander effect. Results : A strong bystander effect was observed in vitro and in vivo. The bystander effect in murine neuroblastoma was dependent on the immune response rather than connexin-mediated gap junction. Neuro-2a/IL-2 treatment enhanced the bystander effect in the HSV-TK/GCV system in murine neuroblastoma model. Conclusion : We conclude that the bystander effect in murine neuroblastoma depends on immune response and is enhanced by neuro-2a/IL-2.

Oxidized Low-density Lipoprotein- and Lysophosphatidylcholine-induced $Ca^{2+}$ Mobilization in Human Endothelial Cells

  • Kim, Moon-Young;Liang, Guo-Hua;Kim, Ji-Aee;Choi, Soo-Seung;Choi, Shin-Ku;Suh, Suk-Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.27-32
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    • 2009
  • The effects of oxidized low-density lipoprotein(OxLDL) and its major lipid constituent lysophosphatidylcholine(LPC) on $Ca^{2+}$ entry were investigated in cultured human umbilical endothelial cells(HUVECs) using fura-2 fluorescence and patch-clamp methods. OxLDL or LPC increased intracellular $Ca^{2+}$ concentration($[Ca^{2+}]_i$), and the increase of $[Ca^{2+}]_i$ by OxLDL or by LPC was inhibited by $La^{3+}$ or heparin. LPC failed to increase $[Ca^{2+}]_i$ in the presence of an antioxidant tempol. In addition, store-operated $Ca^{2+}$ entry(SOC), which was evoked by intracellular $Ca^{2+}$ store depletion in $Ca^{2+}$-free solution using the sarcoplasmic reticulum $Ca^{2+}$ pump blocker, 2, 5-di-t-butyl-l,4-benzohydroquinone(BHQ), was further enhanced by OxLDL or by LPC. Increased SOC by OxLDL or by LPC was inhibited by U73122. In voltage-clamped cells, OxLDL or LPC increased $[Ca^{2+}]_i$ and simultaneously activated non-selective cation(NSC) currents. LPC-induced NSC currents were inhibited by 2-APB, $La^{3+}$ or U73122, and NSC currents were not activated by LPC in the presence of tempol. Furthermore, in voltage-clamped HUVECs, OxLDL enhanced SOC and evoked outward currents simultaneously. Clamping intracellular $Ca^{2+}$ to 1 ${\mu}M$ activated large-conductance $Ca^{2+}$-activated $K^+(BK_{ca})$ current spontaneously, and this activated $BK_{ca}$ current was further enhanced by OxLDL or by LPC. From these results, we concluded that OxLDL or its main component LPC activates $Ca^{2+}$-permeable $Ca^{2+}$-activated NSC current and $BK_{ca}$ current simultaneously, thereby increasing SOC.

A Czochralski Process Design for Si-single Crystal O2 Impurity Minimization with Pulling Rate, Rotation Speed and Melt Charge Level Optimization (Pulling rate, rotation speed 및 melt charge level 최적화에 의한 쵸크랄스키 공정 실리콘 단결정의 O2 불순물 최소화 설계)

  • Jeon, Hye Jun;Park, Ju Hong;Artemyev, Vladimir;Hwang, Seon Hee;Song, Su Jin;Kim, Na Yeong;Jung, Jae Hak
    • Korean Chemical Engineering Research
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    • v.58 no.3
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    • pp.369-380
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    • 2020
  • Most mono-crystalline silicon ingots are manufactured by the Czochralski (Cz) process. But If there are oxygen impurities, These Si-ingot tends to show low-efficiency when it is processed to be solar cell substrate. For making single-crystal Si- ingot, We need Czochralski (Cz) process which melts molten Si and then crystallizing it with seed of single-crystal Si. For melts poly Si-chunk and forming of single-crystalline Si-ingot, the heat transfer plays a main role in the structure of Cz-process. In this study to obtain high-quality Si ingot, the Cz-process was modified with the process design. The crystal growth simulation was employed with pulling rate and rotation speed optimization. Studies for modified Cz-process and the corresponding results have been discussed. The results revealed that using crystal growth simulation, we optimized the oxygen concentration of single crystal silicon by the optimal design of the pulling rate, rotation speed and melt charge level of Cz-process.

Application of Glucuronic Acid with New Cosmetic Active Ingredient (새로운 노화 방지 성분으로서 글루쿠로닉 애씨드의 기능과 화장품 응용)

  • Lee Geun-Soo;Kim Jin-Wha;Lee Chun-Il;Pyo Hyeong-Bae;Lee Kong-Joo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.471-477
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    • 2004
  • Exposure to elevated temperatures, chemical (active oxigen), or physical stress (UV light) induces immediate physiological response, the expression of heat shock proteins in cells. Thus, cells with elevated Heat Shock Protein levels become more tolerant to stress conditions that are otherwise lethal. First, we studied on the new function of glucuronic acid (GA) as preventive material of skin aging. The application of the GA shows significant induction of Heat Shock Protein 70 kDa (HSP 70 kDa) in contrast to cells without it. GA at the concentration which can induce HSP 70 kDa, protects the cell death induced by second stress (heat shock and hydrogen peroxide) in NIH3T3 cells. Second, we studied on in vitro transdermal permeation characteristic of GA through the excised mouse skin. In this study, we compared the skin permeability of GA in water with O/W emulsion. As a result, skin permeation parameters of GA shows lag time 1.2 h, partition coefficient 0.114, permeation flult rate $0.83114 mg/cm^2/h.$ In case of lag time, O/W emulsion containing GA increase 2.48 h. Also, the total accumulation permeation content decreased in contrast to GA solution after 24 h. But it has long-term permeability of glucuronic acid. These results suggest that glucuronic acid could be a good cosmetic active ingredient.

STUDY ON MUTATION OF RAS GENE IN DMBA INDUCED CARCINOMA OF HAMSTER BUCCAL POUCH (DMBA로 유도된 햄스터 협낭암종에서 ras 유전자 변이에 관한 연구)

  • Song, Sun-Chul;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.6
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    • pp.581-590
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    • 2000
  • Alterations in the cellular genome affecting the expression or function of genes controlling cell growth and differentiation are considered to be the main cause of cancer. Over 30 oncogenes can be activated by insertional mutagenesis, single point mutations, chromosomal translocations and gene amplification. The ras oncogenes have been detected in $15{\sim}20%$ of human tumors that include some of the most common forms of human neoplasia and are known to acquire their transforming properties by single point mutations in two domains of their coding sequences, most commonly in codons 12 and 61. The ras gene family consists of three functional genes, N-ras, K-ras and H-ras which encode highly similar proteins of 188 or 189 amino acid residues generically known as P21. ras proteins have been shown to bind GTP and GTP, and possess intrinsic GTPase activity. Experimental study was performed to observe the mutational change of the ras gene family and apply the results to the clinical activity. 36 Golden Syrian Hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek (control side) was treated with mineral oil as the same manner of the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were completely dissected by microdissection and DNA from both tissue were isolated by proteinase K/phenol/chloroform extraction. Segments of the K-ras and H-ras gene were amplified by PCR using the oligonucleotide primers corresponding to the homologous region (codon 12 and 61) of the hamster gene, and then confirmational change of ras genes was observed by SSCP and autosequencing analysis. The results were as follows : 1. Malignant lesion could be found in the experimental side from the experimental six weeks. 2. One hamster among six showed point mutation of the H-ras codon 12($G{\rightarrow}A$ transition) at the experimental 10 and 14 weeks. 3. One of six at 6 weeks, two of six at 8 weeks and one of six at 12 weeks revealed the confirmational change of the H-ras codon 61($A{\rightarrow}T$ transversion). 4. The incidence of point mutation of H-ras codon 12 and 61 were 5.5%(2 of 36) and 11%(4 of 36) respectively. 5. Point mutation of the K-ras could not be seen during the whole experimental period. Form the above results, these findings strongly support the concept that H-ras oncogenes may have the influence of the DMBA induced carcinoma of hamster buccal pouch.

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X-ray Analys is of the Thermotropic Liquid Crystalline Copolyester Poly(1 -phenylethylpphenylene-tere phthalate) (열방성 액정폴리에스터Poly(1-phenylethyl.p-phenyleneterephthalate)의 X-선 결정구조해석)

  • 홍성권
    • Korean Journal of Crystallography
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    • v.2 no.2
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    • pp.13-21
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    • 1991
  • X-ray methods have been used to determine the chain conformation and packing of the thermotropic liquid crystalline copolyester prepared from 50% tarephthaloyl chloride(TPA) and 50% (1-phenylethyl) hydroquinone(PEHQ). The x-ray patterns of annealed melt-spun fibers contain a series of annealed melt-Pointing to a well ordered crystalline structure, despite the random sense(2 or 3-) of the 1-phenylethyl substiuttion on the TPA-hydroquinone backbone. The crystalline fiber is monoclinic with space group P2l and the unit cell has dimensions 11=12.77 A, b=10.17 A (upique axis), c=12.58 h (fiber axis). and β=90.1° and contains TPA-PEHO units of to or chains. The random substitution of 1-phenylethyl groups was modelled by placing these groups at both the 2and 3 positions and giving each a weight of one-hal(. T he structure has been refined by linked a rom least square methods(LALS) against 16 observed and 21 unobserved reflections. and had a final R value of 0.20. Packing of the side chains is effected by staggering adjacent chains along the b axis by approximately c/2, so that the side chains are interleaved. The phenyl-COO and COO-phenyl torsion angles are -6.1 and 65.6, respectively, such that the main chain phenyls are mutually inclined at 59.5 (the ester groups are assumed to be planar). These torsion angles compare very well with those for the model compounds, notably phenylbenzoate, and can be used in future analyses of the structures of more complex random sequence copolyesters.

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Analogues of Hybrid Antimicrobial Peptide, CAMA-P2, Designed with Improved Antimicrobial and Synergistic Activities

  • Jeong, Ki-Woong;Shin, So-Young;Kim, Jin-Kyoung;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.8
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    • pp.2577-2583
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    • 2011
  • We have designed a 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) with high bacterial cell selectivity. CAMA-P2 is an ${\alpha}$-helical antimicrobial peptide designed from a CAMA hybrid peptide and substitution of Gly-Ile-Gly hinge sequence of CAMA to Pro influences the flexibility at central part of CAMA. Based on structure-activity relationships of CAMA peptides, to investigate the effects of the total positive charges on antimicrobial activity of CAMA-P2, the $Ser^{14}{\rightarrow}$Lys analogue (CAMA-syn1) was synthesized. The role of tryptophan at C-terminal ${\alpha}$-helix on its antimicrobial activity as well as synergistic activity was also investigated using $Ser^{14}{\rightarrow}$Lys/$Phe^{18}{\rightarrow}$Trp analogue (CAMA-syn2). Also, we designed CAMA-syn3 by substitution of $Lys^{16}$ located opposite side of substituted $Lys^{14}$ of CAMA-syn1 with Leu residue, resulting in increase of hydrophobicity and amphipathicity of the peptide. All of CAMA-syn analogues showed good antimicrobial activities similar to those of CAMA and CAMA-P2. The CAMA-syn1 and CAMA-syn2 showed low hemolytic activity and cytotoxicity against human keratinocyte Haca-T cells while CAMA-syn3 showed hemolytic activity and cytotoxicity at its MIC value. We then investigated their abilities to act synergistically in combination with the antimicrobial flavonoids and synthetic compounds screened in our laboratory. The results showed that all peptides exhibited synergistic effects with dihydrobinetin, while only CAMA-syn2 exhibited synergistic effects with YKAs3001 against both S. aureus and MRSA, suggesting that Trp residue at C-terminus of CAMA-syn2 may facilitate the polar antibiotic flavonoids and synthetic compounds to permeabilize the membrane. This study will be useful for the development of new antibiotic peptides with potent antimicrobial and synergistic activity but without cytotoxicity.