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Metastatic Axillary Lymph Node Ratio (LNR) is Prognostically Superior to pN Staging in Patients with Breast Cancer -- Results for 804 Chinese Patients from a Single Institution

  • Xiao, Xiang-Sheng;Tang, Hai-Lin;Xie, Xin-Hua;Li, Lai-Sheng;Kong, Ya-Nan;Wu, Min-Qing;Yang, Lu;Gao, Jie;Wei, Wei-Dong;Xie, Xiaoming
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5219-5223
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    • 2013
  • The number of axillary lymph nodes involved and retrieved are important prognostic factors in breast cancer. The purpose of our study was to investigate whether the lymph node ratio (LNR) is a better prognostic factor in predicting disease-free survival (DFS) for breast cancer patients as compared with pN staging. The analysis was based on 804 breast cancer patients who had underwent axillary lymph node dissection between 1999 and 2008 in Sun Yat-Sen University Cancer Center. Optimal cutoff points of LNR were calculated using X-tile software and validated by bootstrapping. Patients were then divided into three groups (low-, intermediate-, and high-risk) according to the cutoff points. Predicting risk factors for relapse were performed according to Cox proportional hazards analysis. DFS was estimated using the Kaplan-Meier method and compared by the log-rank test. The 5-year DFS rate decreased significantly with increasing LNRs and pN. Univariate analysis found that the pT, pN, LNR, molecule type, HER2, pTNM stage and radiotherapy well classified patients with significantly different prognosis. By multivariate analysis, only LNR classification was retained as an independent prognostic factor. Furthermore, there was a significant prognostic difference among different LNR categories for pN2 category, but no apparent prognostic difference was seen between different pN categories in any LNR category. Therefore, LNR rather than pN staging is preferable in predicting DFS in node positive breast cancer patients, and routine clinical decision-making should take the LNR into consideration.

The Effects of SO2 and NH3 on the N2O Reduction with CO over MMO Catalyst (MMO 촉매와 CO 환원제에 의한 N2O 분해에서 SO2 및 NH3 영향 연구)

  • Chang, Kil Sang;You, Kyung-Chang
    • Applied Chemistry for Engineering
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    • v.20 no.6
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    • pp.653-657
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    • 2009
  • Nitrous oxide is a typical greenhouse gas which is produced from various organic or fossil fuel combustion processes as well as chemicals producing plants. $N_2O$ has a global worming potential of 310 times that of $CO_2$ on per molecule basis, and also acts as an ozone depleting material in the stratosphere. However, its removal is not easy for its chemical stability characteristics. Most SCR processes with several effective reducing agents generally require the operation temperature higher than $450^{\circ}C$, and the catalytic conversion becomes decreased significantly when NOx is present in the stream. Present experiments have been performed to obtain basic design data of actual application concerning the effects of $SO_2$ and $NH_3$ on the interim and long term activities of $N_2O$ reduction with CO over the mixed metal oxide (MMO) catalyst derived from a hydrotalcite-like compound precursor. The MMO catalysts used in the experiments, have shown prominent activities displaying full conversions of $N_2O$ near $200^{\circ}C$ when CO is introduced. The presence of $SO_2$ is considered to show no critical behavior as can be met in the $NH_3$ SCR DeNOx systems and the effect of $NH_3$ is considered to play as mere an impurity to share the active sites of the catalysts.

Preparation and Characterization of Small Intestine Submucosa Powder Impregnated Poly(L-lactide) Scaffolds: The Application for Tissue Engineered Bone and Cartilage

  • Khang, Gilson;Rhee, John M.;Shin, Philkyung;Kim, In Young;Lee, Bong;Lee, Sang Jin;Lee, Young Moo;Lee, Hai Bang;Lee, Ilwoo
    • Macromolecular Research
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    • v.10 no.3
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    • pp.158-167
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    • 2002
  • In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

Antiviral Effect of Retro-2.1 against Herpes Simplex Virus Type 2 In Vitro

  • Dai, Wenwen;Wu, Yu;Bi, Jinpeng;Wang, Jingyu;Wang, Shuai;Kong, Wei;Barbier, Julien;Cintrat, Jean-Christophe;Gao, Feng;Jiang, Zhengran;Gillet, Daniel;Su, Weiheng;Jiang, Chunlai
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.849-859
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    • 2018
  • Herpes simplex virus type 2 (HSV-2) infection has been a public health concern worldwide. It is the leading cause of genital herpes and a contributing factor to cervical cancer and human immunodeficiency virus (HIV) infection. No vaccine is available yet for the treatment of HSV-2 infection, and routinely used synthetic nucleoside analogs have led to the emergence of drug resistance. The small molecule $Retro-2^{cycl}$ has been reported to be active against several pathogens by acting on intracellular vesicle transport, which also participates in the HSV-2 lifecycle. Here, we showed that Retro-2.1, which is an optimized, more potent derivative of $Retro-2^{cycl}$, could inhibit HSV-2 infection, with 50% inhibitory concentrations of $5.58{\mu}M$ and $6.35{\mu}M$ in cytopathic effect inhibition and plaque reduction assays, respectively. The cytotoxicity of Retro-2.1 was relatively low, with a 50% cytotoxicity concentration of $116.5{\mu}M$. We also preliminarily identified that Retro-2.1 exerted the antiviral effect against HSV-2 by a dual mechanism of action on virus entry and late stages of infection. Therefore, our study for the first time demonstrated Retro-2.1 as an effective antiviral agent against HSV-2 in vitro with targets distinct from those of nucleoside analogs.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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Synthesis and Crystal Structure of Ag4Br4 Nanoclusters in the Sodalite Cavities of Fully K+-Exchanged Zeolite A (LTA)

  • Lim, Woo-Taik;Choi, Sik-Young;Kim, Bok-Jo;Kim, Chang-Min;Lee, In-Su;Kim, Seok-Han;Heo, Nam-Ho
    • Bulletin of the Korean Chemical Society
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    • v.26 no.7
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    • pp.1090-1096
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    • 2005
  • $Ag_4Br_4$ nanoclusters have been synthesized in about 75% of the sodalite cavities of fully $K^+$-exchanged zeolite A (LTA). An additional KBr molecule is retained in each large cavity as part of a near square-planar $K_4Br^{3+}$ cation. A single crystal of $Ag_{12}$-A, prepared by the dynamic ion-exchange of $Na_{12}$-A with aqueous 0.05 M $AgNO_3$ and washed with $CH_3OH$, was placed in a stream of flowing 0.05 M KBr in $CH_3OH$ for two days. The crystal structure of the product ($K_9(K_4Br)Si_{12}Al_{12}O_{48}{\cdot}0.75Ag_4Br_4$, a = 12.186(1) $\AA$) was determined at 294 K by single-crystal X-ray diffraction in the space group Pm m. It was refined with all measured reflections to the final error index $R_1$ = 0.080 for the 99 reflections for which $F_o\;{\gt}\;4_{\sigma}\;(F_o)$. The thirteen $K^+$ ions per unit cell are found at three crystallographically distinct positions: eight $K^+$ ions in the large cavity fill the six-ring site, three $K^+$ ions fill the eight-rings, and two $K^+$ ions are opposite four-rings in the large cavity. One bromide ion per unit cell lies opposite a four-ring in the large cavity, held there by two eight-ring and two six-ring $K^+$ ions ($K_4Br^{3+}$). Three $Ag^+$ and three $Br^-$ions per unit cell are found on 3-fold axes in the sodalite unit, indicating the formation of nano-sized $Ag_4Br_4$ clusters (interpenetrating tetrahedra; symmetry $T_d$; diameter ca. 7.9 $\AA$) in 75% of the sodalite units. Each cluster (Ag-Br = 2.93(3) $\AA$) is held in place by the coordination of its four $Ag^+$ ions to the zeolite framework (each $Ag^+$ cation is 2.52(3) $\AA$ from three six-ring oxygens) and by the coordination of its four $Br^-$ ions to $K^+$ ions through six-rings (Br-K = 3.00(4) $\AA$).

Peroxidase-Catalyzed Removal of Aromatic Pollutants (Peroxidase에 의한 방향성 유해물질의 제거)

  • 류근갑;박재홍임근호
    • KSBB Journal
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    • v.11 no.6
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    • pp.681-688
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    • 1996
  • In the removal of phenolic precipitates formed by horseradish peroxidase (HRP) and $H_2O_2$ from waste water, the effects of the concentrations of phenolic compounds and $H_2O_2$ on the removal efficiency of various phenols were studied. More than 90% of various phenolic compounds were removed from the aqueous solutions (pH 5-7) by HRP and H2O2. The removal efficiency of phenolic compounds by HRP was reduced to a great extent when the initial concentration of $H_2O_2$ was over 10mM. Furthermore, no phenolic compounds were removed when 50mM of $H_2O_2$ was used. The HRP's turnover number, which indicates the number of phenolic molecules removed per one molecule of HRP, was the largest as 18047 for p-ethoxyphenol while it was the smallest as 1244 for m-chlorophenol when the initial concentrations of phenolic compounds and H2O2 were the same at 1mM. HRP which was separated from the aqueous solution containing phenol and $H_2O_2$ after 24hr of reaction revealed structural changes and diminished activity. The Soret absorbance near 404nm of this HRP sample was decreased to 48% of that of fresh HRP. The values of kcat and kcat/Km of this HRP sample for the oxidation of guaiacol were also reduced to 41% and 51% of those of fresh HRP, respectively. The removals of nonphenolic aromatic compounds such as benzene, ethylbenzene, and toluene (BET) by HRP and $H_2O_2$ were enhanced when phenols were coexisting in the aqueous solutions of BET.

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In vitro antioxidative and anti-inflammatory effects of the compound K-rich fraction BIOGF1K, prepared from Panax ginseng

  • Hossen, Muhammad Jahangir;Hong, Yong Deog;Baek, Kwang-Soo;Yoo, Sulgi;Hong, Yo Han;Kim, Ji Hye;Lee, Jeong-Oog;Kim, Donghyun;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.43-51
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    • 2017
  • Background: BIOGF1K, a compound K-rich fraction prepared from the root of Panax ginseng, is widely used for cosmetic purposes in Korea. We investigated the functional mechanisms of the anti-inflammatory and antioxidative activities of BIOGF1K by discovering target enzymes through various molecular studies. Methods: We explored the inhibitory mechanisms of BIOGF1K using lipopolysaccharide-mediated inflammatory responses, reporter gene assays involving overexpression of toll-like receptor adaptor molecules, and immunoblotting analysis. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to measure the antioxidative activity. We cotransfected adaptor molecules, including the myeloid differentiation primary response gene 88 (MyD88) and Toll/interleukin-receptor domain containing adaptor molecule-inducing interferon-${\beta}$ (TRIF), to measure the activation of nuclear factor (NF)-${\kappa}B$ and interferon regulatory factor 3 (IRF3). Results: BIOGF1K suppressed lipopolysaccharide-triggered NO release in macrophages as well as DPPH-induced electron-donating activity. It also blocked lipopolysaccharide-induced mRNA levels of interferon-${\beta}$ and inducible nitric oxide synthase. Moreover, BIOGF1K diminished the translocation and activation of IRF3 and NF-${\kappa}B$ (p50 and p65). This extract inhibited the upregulation of NF-${\kappa}B$-linked luciferase activity provoked by phorbal-12-myristate-13 acetate as well as MyD88, TRIF, and inhibitor of ${\kappa}B$ ($I{\kappa}B{\alpha}$) kinase ($IKK{\beta}$), and IRF3-mediated luciferase activity induced by TRIF and TANK-binding kinase 1 (TBK1). Finally, BIOGF1K downregulated the NF-${\kappa}B$ pathway by blocking $IKK{\beta}$ and the IRF3 pathway by inhibiting TBK1, according to reporter gene assays, immunoblotting analysis, and an AKT/$IKK{\beta}$/TBK1 overexpression strategy. Conclusion: Overall, our data suggest that the suppression of $IKK{\beta}$ and TBK1, which mediate transcriptional regulation of NF-${\kappa}B$ and IRF3, respectively, may contribute to the broad-spectrum inhibitory activity of BIOGF1K.

Green Tea Polyphenol Epigallocatechine Gallate (EGCG) Prevented LPS-induced BV-2 Micoglial Cell Activation (BV-2 미세아교세포의 활성에 대한 녹차 유래 폴리페놀 EGCG의 억제 효과)

  • Park, Euteum;Chun, Hong Sung
    • Journal of Life Science
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    • v.26 no.6
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    • pp.640-645
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    • 2016
  • Microglial cells are immediately activated in the central nervous system in response to a variety of neuronal environmental changes, such as injuries or inflammation. In addition to the modulation of the intrinsic immune response, a key role of microglial cells is the phagocytosis of dying cells and cellular debris. In this study, the inhibitory effects of epigallocatechine-3-gallate (EGCG), a most abundant and active polyphenol component of green tea, on lipopolysaccharide (LPS)-induced microglial activation are determined. EGCG dose dependently suppressed LPS-induced nitric oxide production and the expression of inducible nitric oxide synthase (iNOS) in BV-2 microglial cells. EGCG are potent LPS-induced inhibitors of several pro-inflammatory cytokine expressions, such as TNF-α and IL-1β, in microglial cells. Furthermore, EGCG generally inhibits the induction of LPS-mediated microglial activation and potently inhibits the phagocytosis of LPS-stimulated BV2 microglia. Although the conditioned media from LPS-stimulated BV-2 cells caused the SN4741 cell death, that from the conditioned media of EGCG pretreated BV-2 cells did not diminish the viability of SN4741 cells. These results suggest EGCG, a green tea polyphenol, could be a promising available molecule for the modulation of harmful microglial activation.