• 제목/요약/키워드: in-vitro

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Effect of ω3-Fatty Acid Desaturase Gene Expression on Invasion and Tumorigenicity in Human Tongue Squamous Cell Carcinoma Cells (인체 혀의 편평세포암 세포에서 ω3-fatty acid desaturase 유전자 발현이 침윤 및 종양형성에 미치는 영향)

  • Hong, Tae-Hwa;Shin, Soyeon;Han, Seung-Hyeon;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.28 no.8
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    • pp.945-954
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    • 2018
  • Omega-3 polyunsaturated fatty acids (${\omega}3$-fatty acid) have been found to possess anticancer properties in a variety of cancer cell lines and animal models, but their effects in human tongue squamous cell carcinomas (SCCs) remain unclear. This study was designed to examine the effect of ${\omega}3$-fatty acid desaturase (fat-1) gene expression on invasion and tumorigenicity in human tongue SCC cells and the molecular mechanism of its action. Docosahexaenoic acid (DHA) treatment inhibited in vitro invasion in a dose-dependent manner. In zymography, matrix metalloproteinase-9 (MMP-9) and Matrix metallopeptidase-2 (MMP-2) activities were reduced, and MMP-9 and MMP-2 promoter activities were inhibited by the DHA treatment. In addition, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) promoter reporter activities were inhibited in SCC-4 and SCC-9 cells after the DHA treatment. To investigate the effect of a high level of endogenous ${\omega}3$ fatty acids, a stable SCC-9 cell line expressing the ${\omega}3$-desaturase gene (fSCC-9sc) was generated. The growth rate and colony-forming capacity of fSCC-9sc were remarkably decreased as compared with those of fSCC-9cc. Likewise, the tumor size and volume of fSCC-9sc implanted into nude mice were significantly inhibited, with increases in the cell death index. Furthermore, a transwell chamber invasion assay showed a reduction in cell invasion of the fSCC-9sc lines when compared with that of the fSCC-9cc line. These findings suggested that fat-1 gene expression inhibited tumorigenicity, as well as invasion in human tongue SCC cells. Thus, utilization of ${\omega}3$ fatty acids may represent a promising therapeutic approach for chemoprevention and the treatment of human tongue SCCs.

SCANNING ELECTRON MICROSCOPIC STUDY ON THE EFFICACY OF ROOT CANAL WALL DEBRIDEMENT OF ROTARY NI-TI INSTRUMENTS WITH DIFFERENT CUTTING ANGLE (엔진 구동형 니켈-타이타늄 합금파일의 절삭각에 따른 근관성형 효과에 관한 전자현미경적 연구)

  • Jeon, In-Soo;Yoon, Tai-Cheol;Park, Seong-Ho;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.27 no.6
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    • pp.577-586
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    • 2002
  • The purpose of this in vitro study was to compare the effects of root canal cleanness following two Ni-Ti rotary instruments with different rake angle. Thirty-six sound, extracted human premolars with single root were randomly divided into three groups. The used rotary instruments were HEROShaper (Group 1, Micro-Mega, Besancon, France, n=12) and ProFile (Group 2, Maillefer, Ballaigues, Switzerland, n=12). Control group (n=12) was only extirpated with barbed broach (Mani, Matsutani Seisakusho Co., Japan) Group 1 & 2 teeth were prepared to a #40/.04 taper at the apex followed by 1 mm using crown-down technique. After canal preparation and frequent irrigation with 5.25% sodium hypochlorite, the roots split longitudinally into a bucco-lingual direction. Root halves were cross-sectioned in apical third portion again. All root specimens were processed for SEM investigation and photographed. Separate evaluations by one endodontist were undertaken for smear layer on prepared walls with a five score-index for each using reference photograph in root halves. The penetration depth of smear layer into dentinal tubules was also estimated in the other halves. Following results were obtained: 1. Smear layer was observed on all the prepared walls with two experimental groups except control group. 2. Smear layer characteristics in two experimental groups; 1) HEROShaper group showed snowy, dusty appearance and were shown open dentinal tubuli on the prepared walls of almost specimens, and the thickness of smear layer covering onto dentinal surfaces was within 1-2 ${\mu}m$ in a few specimens. 2) ProFile group showed shiny, burnished appearance and complete root canal wall covered by a homogenous smear layer with no open dentinal tubuli in all specimens. The penetration of smear layer into dentinal tubules was found in all specimens and the thickness was at 2-4 ${\mu}m$ in all specimens. These results demonstrated that a completely clean root canal could not be achieved regardless of positive or negative rake angle, which is in accordance with the majority of previous studies on root canal cleanliness In conclusion, through irrigation with antibacterial solutions or chelating agents is recommended to remove the smear layer on prepared canal wall in spite of Ni-Ti instrumentation.

Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line (Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향)

  • Lee Su-Min;Lee Hak-Sung;Lee In-Suk;Co Yu-Kyung;Han Hae-Won;Oh Jung-Hwan;Park Jae-Myung;Choi Myung-Gyu;Chung In-Sik
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.906-912
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    • 2004
  • Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

Re-Cloning by Somatic Cell Nuclear Transfer from a Cloned Korean Native Goat (복제 산양(진순이)의 체세포 핵이식에 의한 Re-Cloning에 관한 연구)

  • Jung, S.Y.;Park, H.S.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.89-95
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    • 2007
  • The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.

Effect of Ecklonia cava Water Extracts on Inhibition of IgE in Food Allergy Mouse Model (식품 알레르기 마우스 모델에서 감태 물 추출물의 IgE 분비 억제 효과)

  • Song, Eu-Jin;Lee, Chung-Jo;Kim, Koth-Bong-Woo-Ri;Jung, Ji-Yeon;Kwak, Ji-Hee;Choi, Moon-Kyoung;Kim, Min-Ji;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1776-1782
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    • 2010
  • This research was done to verify the effect of Ecklonia cava water extracts (ECWE) on inhibition of allergic reactions using ovalbumin (OVA)-immunized food allergy mouse model. For in vitro test, $10\sim100{\mu}g$/mL of ECWE and OVA were added to splenocytes obtained from OVA-immunized mice. The significant reduction of IgE antibody level in culture supernatants of splenocytes was shown in ECWE adding group at all tested concentrations. In addition, ECWE decreased IL-4 and IFN-$\gamma$ levels in supernatants of splenocytes. To confirm the effect of ECWE in in vivo test, ECWE was injected to peritoneal cavity of OVA-immunized mice. Subsequently, IgE level was measured in serum and cultured supernatants of splenocytes. As a result, the injection of ECWE (5 and 10 mg/kg BW) significantly attenuated the secretion of IgE antibody in both serum and splenocytes. In conclusion, the present study indicates that ECWE could suppress in a food allergy mouse model through the inhibition of IgE secretion.

The Effects of Majarol on the Blood Pressure and Heart Rate in Rats and Isolated Frog Heart (Majarine 유도체인 majarol의 흰쥐에 있어서 혈압 및 심박동수에 대한 작용과 적출 개구리 심장에 대한 작용)

  • Lee Jong-Hwoa;Park Young-Hyun;Cho Byung-Heon;Kim Yu-Jae;Kim Jong-Bae;Kim Chun-Sook;Cha Young-Deog;Kim Young-Suk
    • The Korean Journal of Pharmacology
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    • v.22 no.1 s.38
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    • pp.34-44
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    • 1986
  • In our previous studies, we had clarified many pharmacological effects of majarine: the bacteriostatic effect in vitro; the potentiation of hypnotic action of alcohol; hypotensive effect in rats and hypothermic effect in mice. This study was undertaken to search for a new antihypertensive drug. Red crystalline was obtained from majarine (which was extracted from Berberis koreana Palibin) by chemical methods. And this crystalline was identified as $C_{19}H_{16}NO_4$ contained one hydroxy group instead of methoxy group of majarine in isoquinoline ring and named 'Majarol' (5,6-Dihydro-9-hydroxy, 10-methoxybenzo-[g]-1,3-benzodioxolo [5,6-a] quinolizinium). We examined the effects of majarol on blood pressure and heart rate in urethane ancsthetized rats and the rate and amplitude of contraction of isolated frog heart. Several drugs: atropine sulfate, diphenhydramine chloride, hexamethonium bromide, phentolamine, epinephrine, propranolol and isoproterenol were used to clarify the mechanism of the hypotensive action of majarol. The results of experimints were as follows; 1. In low dose (0.5-2mg/kg, i.v.), majarol showed a typical transient hypotensive effect and slight decrease in heart rate. In high dose (5-10 mg/kg, i.v.), majarol showed a typical transient and a subsequent prolonged hypotensive effect and a significant prolonged decrease in heart rate was followed. 2. The hypotensive effects of majarol was not abolished by the pretreatments with atropine sulfate, hexamethonium bromide and diphenhydramine. The pretreatment with phentolamine inhibited significantly the hypotensive effects of majarol and the pretreatment wtih majarol blocked markedly the hypertensive effect of epinephrine. The positive chronotropic effect of isoproterenol was not blocked by the pretreatment with majarol. 3. In low dose, majarol increased the amplitude and decreased rate of contraction, but in high dose, majarol inhibited the amplitude and rate of contraction of isolated frog heart.

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Efficacy of Frozen-Thawed ET in Patients with Old Age or Non-Pregnant in Fresh ET Cycles (고령 환자와 신선주기 배아이식에서 임신에 실패한 환자에서 동결-융해 배아이식의 효용성)

  • Choi, Su Jin;Lee, Sun Hee;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.237-243
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    • 2006
  • Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

Evaluation of Sperm Penetration Ability according to Various Morphological Characteristics of Spermatozoa (정자의 여러가지 형태학적 특징에 따른 정자 침투능의 평가)

  • Kim, J.H.;Jung, K.W.;Lew, Y.O.;Kwon, D.J.;Lim, Y.T.;Kim, J.H.;Nha, D.J.;Lee, J.W.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.1
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    • pp.31-41
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    • 1994
  • Morphological estimation of human spermatozoa is complicated by the fact that there is great natural variation in shape. This natural variation in shapes makes it difficult to say which forms are associated with infertility and which are normal variations. Possibly post coital test or in vitro cervical mucus penetration tests will help to clarify this question by showing which sperm are capable of penetration. The purpose of this investigation was performed to assess distribution of various morphological abnormalities according to the ability of sperm to penetrate cervical mucus. The sperm-mucus penetration using hen's egg white as substituting mucus for human cervical mucus was done in 45 fertile men with normal semen analysis and 122 infertile men with abnormal seminal parameters more than one. The female partners of 122 infertile couples showed normal results in the female fundamental test for fertility. Conventional semen analysis was evaluated according to the WHO standard normal(l980). The detailed classification of the abnormal sperm was made according to David et al(l975). The vitality of the sperm samples determined by eosin yellow-nigrosin stainig according to the method of Eliasson(l977). Results were as follw; 1. The patients had significantly lower total sperm count, motility (%), normal morphology (%), viability and total functional sperm fractions(TFSF) than fertile donors. 2. The mean value of sperm penetration distance of the patients(28.69${\pm}$11.02mm) showed significantly lower than fertile donors(37.33${\pm}$5.49mm). And 43/45 fertile donors(95.5%) as well as 57/122 patients(46.7%) had over 30mm in sperm penetration distance respectively. While 2/45 fertile donors(4.5 %) and 65/122 patient(53.3%) had under 30mm in sperm penetration distance respectively. 3. The morphological abnormalities in fertile donors were significantly lower 23.04${\pm}$5.83% (head = 12.89${\pm}$4.98, neck=6.11${\pm}$3.83%, and tail=3.43${\pm}$2.65%), compared to 36.03${\pm}$14. 40% in patients(head = 15.98 8.60%, neck 11.20${\pm}$6.56% and tail=8.70${\pm}$6.55%). Also, 3 types of sperm abnormalities including head, neck and tail were significantly lower in patient than fertile donors, respectively. Both the patients and fertile donors showed higher distribution of sperm with abnormal head than abnormal neck and tail. 4. The mean morphological abnormalities(SP>30mm) of the patients(30.68 11.64%; head = 15.95${\pm}$9.35%, neck=8.14${\pm}$4.21 %, tail=6.56${\pm}$5.64%) were significantly lower compared to patients(40.72${\pm}$15.01 %; head=16.02${\pm}$7.69%, neck 13.89${\pm}$7.82%, tail=1O.58${\pm}$6.75%) under 30mm in sperm penetration distance. Also, both groups over 30mm and under 30mm in sperm penetration showed distance higher distribution of sperm with abnormal head than abnormal neck and tail. The morphological abnormalities of head did not show significant difference but abnormal neck and tail were significant difference between the over 30mm and under 30mm group in sperm penetration distance.

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Identification of Matrix Mineralization-Related Genes in Human Periodontal Ligament Cells Using cDNA Microarray (cDNA microarray에 의한 치주인대세포의 광물화 결절형성에 관여하는 유전자들의 분석)

  • Shin, Jae-Hee;Park, Jin-Woo;Yeo, Shin-Il;Noh, Woo-Chang;Kim, Moon-Kyu;Kim, Jung-Chul;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.447-463
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    • 2007
  • Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.

The Effects of Ethylacetate Fraction of Sanguisorba officinalis L. on Experimentally-induced Acute Gastritis and Peptic Ulcers in Rats (지유가 흰쥐의 급성위염 및 소화성 궤양에 미치는 영향)

  • Lee, Jae-Joon;Choi, Hyun-Suk;Lee, Jeong-Hwa;Jung, Chang-Joo;Lee, Myung-Yul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1545-1552
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    • 2005
  • Sanguisorba officinalis L. has been used as a traditional remedy for arthritis, neuralgia, diarrhea, vomiting, gastric ulcer and gastro-intestinal disorders. This study was performed to investigate the protective effects of ethylacetate fraction of Sanguisorba officinalis L. ethanol extract (SOE) on the gastric mucosal damage and gastric ulcer induced by HCI-ethanol, indomethacin, Shay-ligation method, restraint and water-immersion stress, and cysteamine in rats. The experiment animals were divided into four groups: a negative control group (CON), positive control group (cimetidine 100 mg/kg-CMT or omeprazol 100 mg/kg administrated group-OMT), SOE 200 mg/kg administrated group (SOL) and SOE 400 mg/kg administrated group (SOH), respectively. Rats were given an oral or intraduodenal administration of SOE, and all SOE treatment groups compared with the CON significantly inhibited HCI-ethanol-induced acute gastritis and indomethacin-induced gastric ulcers in dose-dependent manner, of which effects were lower in a positive control drug (CMT). The inhibition rates ($\%$) on the acute gastritis induced by HCI-ethanol and the gastric ulcers induced by indomethacin in rats, 17.66$\%$ and 48.28$\%$ in SOL, 21.71$\%$, and 51.08$\%$ in SOH, and 47.26$\%$ and 58.26$\%$ in CMT compared with CON, respectively, In pylorus ligated rats, the groups of SOE showed decreases in the volume of gastric secretion and acid output. However, no significant differences were observed in the pepsin activity between treated groups . In addition, SOH also depressed gastric ulcers induced by restraint and water-immersion stress and duodenal ulcers induced by cysteamine. These results suggest that Sanguisorba officinalis L. has remarkable antigastric ulcer effects and could be developed as a new antigastric ulcer agents.