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Development of Ecotoxicological Standard Methods using Early Life Stage of Marine Rotifer Brachionus plicatilis and Benthic Copepod Tigriopus japonicus (윤충류 Brachionus plicatilis 및 저서 요각류 Tigriopus japonicus의 초기 생활사를 이용한 해양생태독성시험 방법에 관한 연구)

  • Lee, Seung-Min;Park, Gyung-Soo;Yoon, Sung-Jin;Kang, Young-Shil;Oh, Jeong-Hwang
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.2
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    • pp.129-139
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    • 2008
  • Marine ecotoxicological standard method was applicated using marine rotifer and benthic copepod as primary consumer of marine ecosystem. Marine rotifer, Brachionus plicatilis and benthic copepod, Tigriopus japonicus were designated as standard test species with the endpoints of 24hr neonate mortality (24hr $LC_{50}$) and 48hr population growth (48hr $EC_{50}$) for rotifer, and 48hr nauplius mortality (48hr $LC_{50}$) for benthic copepod. Tests method was referred to those of ASTM (American Society for Testing and Materials) with the replacement of test species which are widely distributed in Korean waters. The two species showed a wide tolerance on salinity ($5{\sim}35\;psu$) and can be easily cultured in small space even they were not as sensitive as in the mortality test using the nauplius of marine invertebrates. However, these species revealed the significant "concentrationresponse relationship" tested with ocean disposal wastes, and reproducibility using cadmium as standard reference material between laboratories. Accordingly, these two species have good potential as test species for marine ecotoxicological test species. Also, we strongly recommend that at least one of these species be included in the test species of "battery test" in marine bioassay.

Quality properties of fermented mugworts and the rapid pattern analysis of their volatile flavor components via surface acoustic wave (SAW) based electronic nose sensor in the GC system (발효 인진쑥과 약쑥의 이화학적 품질특성 및 GC와 SAW센서기반 electronic nose에 의한 향기패턴의 신속분석)

  • Song, Hyo-Nam
    • Food Science and Preservation
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    • v.20 no.4
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    • pp.554-563
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    • 2013
  • The changes in quality properties and nutritional components for two mugworts, namely, Artemisia capillaris Thumberg Artemisiae asiaticae Nakai fermented by Bacillus strains were characterized followed by rapid pattern analysis of volatile flavor compounds through the SAW-based electronic nose sensor in the GC system. After fermentation, the pH has remarkably decreased from 6.0~6.4 to 4.6~5.1 and there has been a slight change in the total soluble solids. The L (lightness) and b (yellowness) values in the Hunter's color system significantly decreased, whilst the a (redness) value increased via fermentation. The HPLC analysis demonstrated that the total amino acids increased in quantity and the essential amino acids were higher in the A. asiaticae Nakai than in the A. capillaris Thumberg, specially with high contents of glutamic and aspartic acid. After fermentation, the monounsaturated fatty acid increased in the A. asiaticae Nakai and the polyunsaturated fatty acids increased in the A. capillaris Thumberg. While the total polyphenol contents have not been affected by fermentation, the total sugar contents have dramatically decreased. Scopoletin, which is one of the most important index components in mugworts, was highly abundant in the A. capillaris Thumberg; however, it was not detected in the A. asiaticae Nakai. Small pieces of plant tissue in the surface microstructure were found in the fermented mugworts through the use of the scanning electron microscope (SEM). Volatile flavor compounds via electronic nose showed that the intensity of several peaks has increased and additional seven flavor peaks have been produced after fermentation. The VaporPrintTM images demonstrated a notable difference in flavors between the A. asiaticae Nakai and A. capillaris Thumberg, and the fermentation enabled the mugworts to produce subtle differences in flavor.

A Study of Fish Community on Up and Downstream of Hwabuk Dam Under Construction in the Upper Wie Stream. (위천 상류에 건설 중인 화북댐 상 하류 어류군집에 관한 연구)

  • Seo, Jin-Won;Kim, Hee-Sung
    • Korean Journal of Ecology and Environment
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    • v.42 no.2
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    • pp.260-269
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    • 2009
  • Hwabuk Dam has been under construction to reduce flood damage in Nakdong River watershed and to supply stable water for middle area of Gyeongbuk Province. Therefore, fish investigation in up and downstream of the dam was conducted from 2004 to 2008 in order to determine any negative effect on fish community due to dam construction and to use as fundamental data for conserving species diversity and maintaining stream health. According to data analysis on water quality, temperature, dissolved oxygen, pH, suspended solids, and total E-coli had seasonal variation, but they did not significantly differ in sites. However, biological and chemical oxygen demand, chlorophyll-a, nitrogen, and phosphorus representing organic matter and nutrient concentration were higher in upper site and decreased to lower site so that they differed by site. Concentration of arsenic among the heavy metals was less than 0.05 mg $L^{-1}$, which is regulated for protection of human health in water quality standard, except for 0.092 mg $L^{-1}$ in June 2005. During the study period, the total number of fish caught from the 6 sites was 10,263 representing 7 families 19 species. Among them, dominant and subdominant species were Korean chub (Zacco koreanus, 62.5%) and Chinese minnow (Rhynchocypris oxycephalus, 10.6%) which inhabit mostly in mid and upper streams, Korea. Among the 19 species, Korean endemic species were 9 species (47.4%) including Korean slender gudgeon (Squalidus gracilis majimae), Korean dark sleeper (Odontobutis platycephala), and Korean shiner (Coreoleuciscus splendidus). There was several individuals of the $1^{st}$-class endangered species, Naktong nose loach (Koreocobitis nahtongensis), caught in 2005${\sim}$2007, and no introduced species of fish was found in entire sampling period. According to result of community analysis, dominance index decreased toward lower site, but diversity and richness indices increased toward lower site. The equation of length-weight relationship on the dominant species was TW=0.000003$(TL)^{3.2603}$. The parameter b in the equation was greater than 3.0 indicating good nutritional condition in the populations. Compared to populations of Korean chub in other streams, the population in Hwabuk Dam watershed had higher mean of condition factor by size indicating better growth rate. With fish fauna and multi-metric health assessment model in each sampling attempt, index of biotic integrity (IBI) was evaluated and it resulted mostly in good (26${\sim}$35) and excellent (36${\sim}$40) condition in all sites, and the mean of IBI was the highest in site 5. The results indicate that it is very important to study not only environmental impact assessment with fish composition but also stream health assessment in order to conserve healthy aquatic ecosystem.

Anti-inflammatory Effect of Ethanol Extract from Sargassum fulvellum on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀조직에 대한 참모자반 (Sargassum fulvellum) 에탄올 추출물의 항염증 효과)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Ahn, Na-Kyung;Choi, Yeon-Uk;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1158-1165
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    • 2014
  • This study investigated the anti-inflammatory effects of Sargassum fulvellum ethanol extract (SFEE) on the lipopolysaccharide (LPS)-induced inflammatory response. SFEE remarkably suppressed production of NO and pro-inflammatory cytokines (IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ at 50 and $100{\mu}g/mL$. There were no cytotoxic effects on proliferation of macrophages treated with SFEE compared to the control. SFEE reduced expression of iNOS and COX-2 proteins in a dose-dependent manner. The formation of edema in mouse ears was reduced at the highest dose tested compared to the control. Moreover, in the acute toxicity test, no mortality occurred in mice administered 5,000 mg/kg body weight of SFEE over the 2-week observation period. These results suggest that SFEE may have significant effects on inflammatory factors and be a potential anti-inflammatory therapeutic material.

Improving Effect of a Combined Extract of Rhei Rhizoma and Glycyrrhizae Rhizoma through Anti-oxidative Stress in Reflux Esophagitis rats (대황 감초 복합추출물의 항산화 효과를 통한 역류성 식도염 개선 효과)

  • Kim, MinYeong;Shin, YuOck;Lee, JooYoung;Lee, AhReum;Shin, SungHo;Kwon, OJun;Seo, BuIl;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.30 no.4
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    • pp.37-44
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    • 2015
  • Objectives : The present study was designed to evaluate the anti-inflammatory and anti-oxidative stress activities through regulation of Nrf2-mediated genes by Rhei rhizoma and Glycyrrhiza rhizoma combined extract (RGE) in reflux esophagitis.Methods : The antioxidant activity of RGE in vitro was measured in terms of radical scavenging capacity such as DPPH and ABTS. RGE was administered at 350 mg/kg body weight prior to induction of reflux esophagitis. Reflux esophagitis was induced that tied the pylorus and the transitional junction between the forestomach and the corpus in Sprague-Dawley rats.Results : RGE scavenged DPPH and ABTS effectively and IC50of RGE each were 4.9 μg/ml and 45.6 μg/ml. Our results show that RGE administration markedly ameliorated mucosal damage upon histological evaluation. In serum and esophagus tissue, RGE significantly suppressed the oxidative stress biomarkers. Reflux esophagitis induced rats exhibited down-regulation of antioxidant-related proteins in the esophagus; however, the levels with treatment of RGE were significantly higher than those of vehicle reflux esophagitis rats. RGE treatment caused significant reductions in activation of NF-κB transcription factor. Thus, RGE significantly exhibited potent anti-inflammatory activities by suppressing the protein expression levels of pro-inflammatory proteins such as COX-2 and iNOS and inflammatory cytokines such as TNF-αin the esophagus tissue.Conclusions : Reflux esophagitis caused considerable levels of oxidative stress in the esophageal mucosa and the administration of RGE reduced the esophageal mucosa damage through the regulation of Nrf2 and NF-κB pathways. Our findings can considered as supplementary therapy in the prevention or treatment of reflux esophagitis.

Selection of Acifluorfen-tolerant Eastern Black Nightshade (Solanum ptycanthum Dun) and the Expression of This Tolerance in Regenerated Plants and Their Progeny (제초제 Acifluorfen 저항성 세포주 선발 및 분화된 식물체와 그 후대에서의 저항성 발현)

  • Chang Yeon Yu;John B. MASIUNG
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.151-156
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    • 1994
  • Acifluorfen-tolerant cell lines of S, ptycanthum were isolated by stepwise selection using suspension culture. Growth of unselected line was completely inhibited at $0.5\;\mu\textrm{M}$, while some selected lines grew at $8\;\mu\textrm{M}$ acifluorfen. After subculturing on acifluorfen-free medium for 4 passages, six of the eleven cell lines screened and maintained their tolerance to $2\;\mu\textrm{M}$ acifluorfen. The regeneration capacity of selected cell lines in Solanum ptycanthum differed depending on the tell line. The acifluorfen tolerance of the somarclones regenerated from acifluorfen-tolerant cell lines differed depending on the somarclone. When plants were heated with $16\;\mu\textrm{M}$ acifluorfen, unselected control plane had over 75% phytotoxicity Many selected cell lines had less phytotoxicity than the seed-grown control plants. Tolerance to acifluorfen was inherited to the self-pollinated progenies. The inheritance patterns differed depending on the clone. Acifluorfen tolerance was inherited as a semidominant trait. Other segregation patterns were also observed. acifluorfen tolerance was recessive and acifluorfen sensitivity was dominant.

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The Effects of LR3 and SP6 Acupuncture on Renal Damage in Streptozotocin-induced Diabetic Mice (태충·삼음교의 침 자극이 Streptozotocin으로 유발된 당뇨쥐의 신장 손상에 미치는 영향)

  • Lee, Cho In;Lee, Hyun Jong;Lee, Yun Kyu;Lim, Seong Chul;Kim, Jae Soo
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.41-51
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    • 2015
  • Objectives : This study was performed to investigate the effects of $LR_3$ and $SP_6$ acupuncture on renal damage in streptozotocin(STZ)-induced diabetic mice. Methods : ICR male mice were stabilized for a week and divided into four groups: a normal mice group(N), no-acupuncture diabetic mice group(Control), $LR_3$ acupuncture diabetic mice group($LR_3$), and $SP_6$ acupuncture diabetic mice group($SP_6$). Diabetes was experimentally induced by intraperitoneal injection of STZ(150 mg/kg) in citrate buffer(pH 4.5). For two weeks, $LR_3$ and $SP_6$ acupunctures were administered bilaterally at each point once a day. After two weeks, the animals' weight was measured and they underwent a laparotomy. Serum glucose and blood urea nitrogen(BUN) were measured from the blood taken from the heart. We measured glucose, reactive oxygen species(ROS), peroxynitrite($ONOO^-$) and thiobarbituric acid reactive substances(TBARS) in the kidney and compared expression levels of superoxide dismutases(SOD), glutathione peroxidase(GPx), nuclear factor-kappa B(NF-${\kappa}B$), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase(iNOS) and Interleukin-1 beta(IL-$1{\beta}$). Results : BUN significantly decreased in $LR_3$, $SP_6$ compared to the control group. $LR_3$ showed significantly decreased glucose compared to the control group. $LR_3$, $SP_6$ significantly decreased in ROS and $ONOO^-$ compared to the control group. $LR_3$ significantly decreased in TBARS compared to the control group. $SP_6$ significantly increased in expressions of SOD-1, catalase, and GPx compared to the control group. $LR_3$, $SP_6$ significantly decreased in COX-2 compared to the control group. $SP_6$ significantly decreased in IL-$1{\beta}$ compared to the control group. Conclusions : This study suggests that $LR_3$ acupuncture may be effective in controlling glucose and lipid peroxidation and that $SP_6$ acupuncture may have anti-oxidative and anti-inflammatory effects on renal damage in STZ-induced diabetic mice.

Some Observations on the Organelles Participating in the Biliary Excretion in the Hepatocyte of the Biligrafin Injected Mouse (Biligrafin 투여 마우스 간세포의 미세구조적 및 세포화학적 연구)

  • Kim, Hyang;Shin, Young-Chul
    • Applied Microscopy
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    • v.23 no.2
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    • pp.53-77
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    • 1993
  • In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

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Effect of Concentration and Exposure Duration of FBS on Parthenogenetic Development of Porcine Follicular Oocytes

  • Kim, Hyun-Jong;Cho, Sang-Rae;Choe, Chang-Yong;Choi, Sun-Ho;Son, Dong-Soo;Kim, Sung-Jae;Sang, Byung-Don;Han, Man-Hye;Ryu, Il-Sun;Kim, In-Cheul;Kim, Il-Hwa;Lee, Woon-Kyu;Im, Kyung-Soon
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.245-249
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    • 2007
  • The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.

Protective Effects of the Ethanol Extract of Viola tianshanica Maxim against Acute Lung Injury Induced by Lipopolysaccharides in Mice

  • Wang, Xue;Yang, Qiao-Li;Shi, Yu-Zhu;Hou, Bi-Yu;Yang, Sheng-Qian;Huang, Hua;Zhang, Li;Du, Guan-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1628-1638
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    • 2017
  • Viola tianshanica Maxim, belonging to the Violaceae plant family, is traditionally used in Uighur medicine for treating pneumonia, headache, and fever. There is, however, a lack of basic understanding of its pharmacological activities. This study was designed to observe the effects of the ethanol extract (TSM) from Viola tianshanica Maxim on the inflammation response in acute lung injury (ALI) induced by LPS and the possible underlying mechanisms. We found that TSM (200 and 500 mg/kg) significantly decreased inflammatory cytokine production and the number of inflammatory cells, including macrophages and neutrophils, in bronchoalveolar lavage fluid. TSM also markedly inhibited the lung wet-to-dry ratio and alleviated pathological changes in lung tissues. In vitro, after TSM ($12.5-100{\mu}g/ml$) treatment to RAW 264.7 cells for 1 h, LPS ($1{\mu}g/ml$) was added and the cells were further incubated for 24 h. TSM dose-dependently inhibited the levels of proinflammatory cytokines, such as NO, $PGE_2$, $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$, and remarkably decreased the protein and mRNA expression of $TNF-{\alpha}$ and IL-6 in LPS-stimulated RAW 264.7 cells. TSM also suppressed protein expression of $p-I{\kappa}Ba$ and p-ERK1/2 and blocked nuclear translocation of $NF-{\kappa}B$ p65. The results indicate that TSM exerts anti-inflammatory effects related with inhibition on $NF-{\kappa}B$ and MAPK (p-ERK1/2) signaling pathways. In conclusion, our data demonstrate that TSM might be a potential agent for the treatment of ALI.