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Partial Sequencing and Characterization of Porcine DNA Methyltransferase I cDNA

  • Lee, Y.Y.;Kim, M.S.;Park, J.J.;H.Y. Kang;Y.M. Chang;Yoon, J.T.;K.S. Min
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.84-84
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    • 2003
  • DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. DNA methylation is a highly plastic and critical component of mammalian development The DNA methyltransferases (Dnmts) are responsible for the generation of genomic methylation patterns, which lead to transcriptional silencing. The maintenance DNA methyltransferase enzyme, Dnmt 1, and the de novo methyltransferase, Dnmt3a and Dnmt3b, are indispensable for development because mice homozygous for the targeted disruption of any of these genes are not viable. The occurrence of DNA methylation is not random, and it can result in gene silencing The mechanisms underlying these processes are poorly understood. It is well established that DNA methylation and histone deacetylation operate along a common mechanistic pathway to repress transcription through the action of methyl-binding domain proteins (MBDs), which are components of, or recruit, histone deacetylase (HDAC) complexes to methylated DNA. As a basis for future studies on the role of the DNA-methyl-transferase in porcine development, we have isolated and characterized a partial cDNA coding for the porcine Dnmt1. Total RNA of testis, lung and ovary was isolated with TRlzol according to the manufacture's specifications. 5 ug of total RNA was reverse transcribed with Super Script II in the presence of porcine Dnmt 1 specific primers. Standard PCRs were performed in a total volume of 50 ul with cDNA as template. Two DNA fragmenets in different position were produced about 700bp, 1500bp and were cloned into pCR II-TOPO according to the manufacture's specification. Assembly of all sequences resulted in a cDNA from 158bp of 5'to 4861bp of 3'compare with the known human maintenance methyltransferase. Now, we are cloning the unknown Dnmt 1 region by 5'-RACE method and expression of Dnmt 1 in tissues from adult porcine animals.

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The responsibility of C-terminal domain in the thermolabile haemolysin activity of Vibrio parahaemolyticus and inhibition treatments by Phellinus sp. extracts

  • Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.204-215
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    • 2023
  • The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

Assessment of Phenolic Content, Saponin Content, and Antioxidant Activities in Gray, Red, and White Adzuki Bean Germplasm: A Multivariate Analysis

  • Kebede Taye Desta;Hyemyeong Yoon;Myoung-Jae Shin;Sukyeung Lee;Xiaohan Wang;Yu-Mi Choi;Young-ah Jeon;YoungKwang Ju;JungYoon Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.3
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    • pp.147-166
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    • 2023
  • Seed color is controlled by several genes and is a key trait in determining the metabolite content and biological activities of legume genotypes. In this study, 296 adzuki bean accessions, including 159 grey, 99 red, and 38 white adzuki beans, were grown in Korea. Variations in total phenolic content (TPC), total saponin content (TSC), DPPH scavenging activity, ABTS•+ scavenging activity, and ferric reducing antioxidant power (FRAP) were assessed and were reported to be in the ranges of 1.52-8.24 mg GAE/g, 14.36-114.22 mg DE/g, 0.23-12.84 mg AAE/g, 1.05-17.66 mg TE/g, and 0.59-13.14 mg AAE/g, respectively, with a wide variation across adzuki beans. Except for DPPH scavenging activity, the average values declined in the order gray > red > white adzuki beans, each demonstrating a significant variation (p < 0.05). White adzuki beans, which showed low metabolite content and antioxidant activity, were clearly separated from the gray and red genotypes using principal component and hierarchical cluster analyses. Moreover, TPC, TSC, and antioxidant activities were strongly correlated, regardless of seed color. Overall, the diversity of the TPC, TSC, and antioxidant activity in a broad population of adzuki bean genotypes was determined. Furthermore, this study found that seed color variation in adzuki beans had a significant effect on the metabolite content and antioxidant activity. Superior accessions with high levels of TPC, TSC, and antioxidant activity were also discovered and could be used for functional plant breeding and human consumption. The findings of this study may be useful for understanding the relationship between seed coat color and metabolite concentration in adzuki beans, paving the way for molecular-level analyses.

T2 Mapping with and without Fat-Suppression to Predict Treatment Response to Intravenous Glucocorticoid Therapy for Thyroid-Associated Ophthalmopathy

  • Linhan Zhai;Qiuxia Wang;Ping Liu;Ban Luo;Gang Yuan;Jing Zhang
    • Korean Journal of Radiology
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    • v.23 no.6
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    • pp.664-673
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    • 2022
  • Objective: To evaluate the performance of baseline clinical characteristics and pretherapeutic histogram parameters derived from T2 mapping of the extraocular muscles (EOMs) in the prediction of treatment response to intravenous glucocorticoid (IVGC) therapy for active and moderate-to-severe thyroid-associated ophthalmopathy (TAO) and to investigate the effect of fat-suppression (FS) in T2 mapping in this prediction. Materials and Methods: A total of 79 patients clinically diagnosed with active, moderate-to-severe TAO (47 female, 32 male; mean age ± standard deviation, 46.1 ± 10 years), including 43 patients with a total of 86 orbits in the responsive group and 36 patients with a total of 72 orbits in the unresponsive group, were enrolled. Baseline clinical characteristics and pretherapeutic histogram parameters derived from T2 mapping with FS (i.e., FS T2 mapping) or without FS (i.e., conventional T2 mapping) of EOMs were compared between the two groups. Independent predictors of treatment response to IVGC were identified using multivariable analysis. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive performance of the prediction models. Differences between the models were examined using the DeLong test. Results: Compared to the unresponsive group, the responsive group had a shorter disease duration, lower kurtosis (FS-kurtosis), lower standard deviation, larger 75th, 90th, and 95th (FS-95th) T2 relaxation times in FS mapping and lower kurtosis in conventional T2 mapping. Multivariable analysis revealed that disease duration, FS-95th percentile, and FS-kurtosis were independent predictors of treatment response. The combined model, integrating all identified predictors, had an optimized area under the ROC curve of 0.797, 88.4% sensitivity, and 62.5% specificity, which were significantly superior to those of the imaging model (p = 0.013). Conclusion: An integrated combination of disease duration, FS-95th percentile, and FS-kurtosis was a potential predictor of treatment response to IVGC in patients with active and moderate-to-severe TAO. FS T2 mapping was superior to conventional T2 mapping in terms of prediction.

Comparison of Kinematics and Myoelectrical Activity during Deadlift, with and without Variable Banded Resistance, in Healthy, Trained Athletes

  • Everett B. Lohman;Mansoor Alameri;Fulden Cakir;Chih Chieh Chia;Maxine Shih;Owee Mulay;Kezia Marceline;Simran Jaisinghani;Gurinder Bains;Michael DeLeon;Noha Daher
    • Physical Therapy Rehabilitation Science
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    • v.13 no.1
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    • pp.53-70
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    • 2024
  • Background: The conventional deadlift is a popular exercise for enhancing trunk, core, and lower extremity strength. However, its use in sports medicine is constrained by concerns of lumbar injuries, despite evidence supporting its safety and rehabilitative benefits. To optimize muscle activation using resistive bands in variable resistance therapy, we explored their feasibility in the deadlift. Design: Comparative experimental design Methods: Surface electromyography recorded muscle activity in the trunk and lower extremities during lifting, with normalization to the isometric Floor Lift using Maximal Voluntary Contraction. Kinematics were measured using inclinometer sensors to track hip and trunk sagittal plane angles. To prevent fatigue, each subject only used one of the three pairs of bands employed in the study. Results: Our study involved 45 healthy subjects (mean age: 30.4 ± 6.3 years) with similar baseline characteristics, except for years of lifting and strength-to-years-of-lifting ratio. Various resistance band groups exhibited significantly higher muscle activity than conventional deadlifts during different phases. The minimal resistance band group had notably higher muscle activity in the trunk, core, and lower extremity muscles, particularly in the end phase. The moderate resistance band group showed increased muscle activity in the mid-and end-phases. The maximum resistance band group demonstrated greater muscle activity in specific muscles during the early phase and overall higher activity in all trunk and lower extremity muscles in the mid and end phases of the deadlift (p<0.05). Conclusion: Our findings provide valuable insights into muscle activation with various resistance bands during deadlift exercise in clinical and gym settings. There appears to be a dose-response relationship between increased resistance bandwidth, external load, myoelectric activation, and range.

Ischemic Burden Assessment Using Single Photon Emission Computed Tomography in Single Vessel Chronic Total Occlusion of Coronary Artery

  • Yong-Hoon Yoon;Sangwon Han;Osung Kwon;Kyusup Lee;Ju Hyeon Kim;Junghoon Lee;Tae oh Kim;Jae-Hyung Roh;Pil Hyung Lee;Soo-Jin Kang;Jae-Hwan Lee;Young-Hak Kim;Cheol Whan Lee;Dae Hyuk Moon;Seung-Whan Lee
    • Korean Circulation Journal
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    • v.52 no.2
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    • pp.150-161
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    • 2022
  • Background and Objectives: Studies evaluating the nature of ischemic burden of chronic total occlusion (CTO) vessels are still lacking. Methods: A total of 165 patients with single vessel CTO >2.5 mm in an epicardial coronary artery who underwent single photon emission computed tomography (SPECT) were enrolled in the study. Ischemic burden was calculated with the use of semi-quantitative SPECT analysis, and was defined as the summed difference score (SDS) divided by the maximal limit of the score (=SDS/68). Results: The mean age of the participants was 59.5 years and the CTO of the left anterior descending coronary artery (LAD), left circumplex coronary artery (LCX), and right coronary artery (RCA) accounted for 93 (56.4%), 18 (10.9%), and 54 (32.7%) patients, respectively. The median ischemic burden of the total population was 8.8%, and it was highest in the LAD CTO (10.3%) compared with the LCX (5.9%) and RCA CTO (5.9%, p<0.001). High-ischemic burden (ischemic burden >10%) was observed in 66 patients (40.0%), and in 47 patients (50.5%) of the LAD CTO. Ischemic burden was different according to the CTO location only in LAD CTO. The statistically significant predictors for high-ischemic burden were hypertension, baseline ejection fraction >45%, LAD CTO, proximal CTO location, and de novo CTO. Japanese-CTO score and Rentrop scale collateral grade were not associated with high-ischemic burden. Conclusions: Only 40% of patients with single vessel CTO had ischemic burden >10%. For CTO vessels, measurement of ischemic burden using SPECT prior to revascularization may be helpful in identifying beneficial subjects.

Effects of a Blend of Prunus Mume Extract as an Alternative to Antibiotics on Growth Performance, Activity of Digestive Enzymes and Microflora Population in Broiler Chickens (항생제 대체 사료첨가제로서 매실추출 혼합물이 육계의 성장, 소화 효소 활성도 및 장내 미생물 균총에 미치는 영향)

  • Ko, Y.H.;Yang, H.Y.;Kang, S.Y.;Kim, E.S.;Jang, I.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.611-620
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    • 2007
  • The current study was designed to define whether a blend of prunus mume extract(25%) containing lactic acid(75%) and grape seed extract(10ppm) could affect in vitro antimicrobial activity and growth performance, intestinal microflora, plasma biochemical profiles and digestive enzymes activities in broiler chickens. In paper disc agar diffusion test, we clearly observed antimicrobial activity against E. coli in response to prunus mume extract or a blend of prunus mume extract. For in vivo test, a total of ninety six 3-d-old male broiler chicks were assigned to basal diet(CON), basal diet supplemented with antibiotics (ANTI) and 0.5% a blend of prunus mume extract(PRNUS) until 35 days of age. Throughout the entire experimental period(3-35 days), there were no differences in BW and FCR between the birds fed the basal diet with antibiotics and the diet supplemented with a blend of prunus mume. However, ANTI group showed a significant increase in BW and total gain compared to CON group. The weights of digestive organs such as the pancreas and mucosal tissues were not affected by dietary treatments. There was no difference in plasma levels of glucose, cholesterol, AST and ALT activity. However, triglyceride in plasma increased(P<0.05) in the birds fed the diet supplemented with 0.5% a blend of prunus mume extract compared to those fed antibiotics supplemented diet. The activities of pancreatic trypsin and amylase, and intestinal hydrolase including disaccharidase were not affected by dietary treatment. The colony forming units(CFU) of lactobacillus in the lower ileal-cecum of the birds fed the diet supplemented with a blend of prunus mume extract was significantly(P<0.05) higher than that of birds fed antibiotic supplemented diet without affecting the CFU of E. coli. In conclusion, the birds fed the diet supplemented a blend of prunus mume as an alternative to antibiotics showed a similar growth performance and an significant increase in lactobacillus population compared with the birds fed basal and antibiotics supplemented diets.

Nutritional Characteristics and Stability in Cell of the Yac-Sun Tea for Caronary Heart Disease (관상동맥 질환의 예방을 위한 약선차의 식품영양학적 구성 및 안전성 평가)

  • Kim, Woon-Ju;Cho, Hwa-Eun;Park, Sung-Hye
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.219-225
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    • 2007
  • This study was performed to provide basic ideas as understanding and application for oriental medicinal cuisine (Yak-Sun). To develop medicinal cuisine, it is necessary to grasp the theoretical system. And to develop medicinal cuisine for health enhancement, it is also required not only to consider constitutions but also to suggest the need of knowledge for moderation in terms of regimen along with the theory of oriental medicine. Also to develop medicinal cuisine according to the perspective of oriental medicinal theory, what should be taken into account is not only the understanding of the characteristics of food materials, but also the properties of them that the theory of oriental medicine. Lastly the scientific effect of the medicinal cuisine which is developed according to the oriental medicinal theory. And it is believed to De essential for the government to make effects to set a standard and laws to validate the medicinal effects and the process of assessment so that the systematic development can be encouraged, and to prepare guidance to food development for national health improvement. This research was planned and executed to evaluate how the composition of Yak-sun(oriental diet therapy) can effect health conditions of people who are suffering from diet-related diseases like cardiovascular related disease. by taking Yak-sun in a form of nutritional supplement with our daily meals. We produced Yak-sun tea with Mansam, Hwanggi, Tanggi and Paekchak and observed nutritional composition. We concluded that we could apply the components not only in a form of tea, but also in other forms of various food. The information we received from this conclusion will be a basic information on how we can apply oriental medicinal resources into other food and will also be a steppingstone for medicinal herbs to step foot in the field of functional food research, which already draws sizable attention world-wide.

Geological Characteristics of Extra Heavy Oil Reservoirs in Venezuela (베네주엘라 초중질유 저류층 지질 특성)

  • Kim, Dae-Suk;Kwon, Yi-Kyun;Chang, Chan-Dong
    • Economic and Environmental Geology
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    • v.44 no.1
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    • pp.83-94
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    • 2011
  • Extra heavy oil reservoirs are distributed over the world but most of them is deposited in the northern part of the Orinoco River in Venezuela, in the area of 5,500 $km^2$, This region, which has been commonly called "the Orinoco Oil Belt", contains estimated 1.3 trillion barrels of original oil-in-place and 250 billion barrels of established reserves. The Venezuela extra heavy oil has an API gravity of less than 10 degree and in situ viscosity of 5,000 cP at reservoir condition. Although the presence of extra heavy oil in the Orinoco Oil Belt has been initially reported in the 1930's, the commercial development using in situ cold production started in the 1990's. The Orinoco heavy oil deposits are clustered into 4 development areas, Boyaco, Junin, Ayachoco, and Carabobo respectively, and they are subdivided into totally 31 production blocks. Nowadays, PDVSA (Petr$\'{o}$leos de Venzuela, S.A.) makes a development of each production block with the international oil companies from more than 20 countries forming a international joint-venture company. The Eastern Venezuela Basin, the Orinoco Oil Belt is included in, is one of the major oil-bearing sedimentary basins in Venezuela and is first formed as a passive margin basin by the Jurassic tectonic plate motion. The major source rock of heavy oil is the late Cretaceous calcareous shale in the central Eastern Venezuela Basin. Hydrocarbon materials migrated an average of 150 km up dip to the southern margin of the basin. During the migration, lighter fractions in the hydrocarbon were removed by biodegradation and the oil changed into heavy and/or extra heavy oil. Miocene Oficina Formation, the main extra heavy oil reservoir, is the unconsolidated sand and shale alternation formed in fluvial-estuarine environment and also has irregularly a large number of the Cenozoic faults induced by basin subsidence and tectonics. Because Oficina Formation has not only complex lithology distribution but also irregular geology structure, geological evolution and characteristics of the reservoirs have to be determined for economical production well design and effective oil recovery. This study introduces geological formation and evolution of the Venezuela extra heavy oil reservoirs and suggest their significant geological characteristics which are (1) thickness and geometry of reservoir pay sands, (2) continuity and thickness of mud beds, (3) geometry of faults, (4) depth and geothermal character of reservoir, (5) in-situ stress field of reservoir, and (6) chemical composition of extra heavy oil. Newly developed exploration techniques, such as 3-D seismic survey and LWD (logging while drilling), can be expected as powerful methods to recognize the geological reservoir characteristics in the Orinoco Oil Belt.

Decitabine in the Treatment of Acute Myeloid Leukemia and Myelodysplastic Syndromes, Which Combined with Complex Karyotype Respectively

  • Gao, Su;Li, Zheng;Fu, Jian-Hong;Hu, Xiao-Hui;Xu, Yang;Jin, Zheng-Ming;Tang, Xiao-Wen;Han, Yue;Chen, Su-Ning;Sun, Ai-Ning;Wu, De-Pei;Qiu, Hui-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6627-6632
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    • 2015
  • Background: We conducted a study exploring the clinical safety and efficacy of decitabine in patients with acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS), combined with a complex karyotype. Materials and Methods: From April 2009 to September 2013, a total of 35 patients with AML/MDS combined with a complex karyotype diagnosed in the First Affiliated Hospital of Soochow University were included for retrospective analysis. All patients were treated with decitabine alone ($20mg/m^2$ daily for 5 days) or combination AAG chemotherapy (Acla 20mg qod*4d, Ara-C $10mg/m^2$ q12h*7d, G-CSF $300{\mu}g$ qd, the dose of G-CSF adjusted to the amount in blood routinely). Results: In 35 patients, 15 exhibited a complete response (CR), and 6 a partial response (PR), the overall response rate (CR+PR) being 60% (21 of 35). Median disease-free survival was 18 months and overall survival was 14 months. In the 15 MDS patients with a complex karyotype, the CR rate was 53.3% (8 of 15); in 20 AML patients with complex karyotype, the overall response rate was 65% (13 of 20). The response rate of decitabine alone (22 cases) was 56.5% (13 of 22), while in the combination chemotherapy group (13 cases), the effective rate was 61.5% (8 of 13)(P>0.05). There are 15 patients with chromosome 7 aberration, after treatment with decitabine, 7 CR, 3 PR, overall response rate was 66.7% (10 of 15). Of 18 patients with 3 to 5 kinds of chromosomal abnormalities, 66.7% demonstrated a response; of 17 with more than 5 chromosomal abnormalities, 52.9% had a response. In the total of 35 patients, with one course (23 patients) and ${\geq}$two courses (12 patients), the overall response rate was 40.9% and 92.3% (P<0.05). Grade III to IV hematological toxicity was observed in 27 cases (75%). Grade III to IV infections were clinically documented in 7 (20%). Grades I to II non-hematological toxicity were infections (18 patients), haematuria (2 patients), and bleeding (3 patients). With follow-up until September 2013, 7 patients were surviving, 18 had died and 10 were lost to follow-up. In the 6 cases who underwent allogeneic hematopoietic stem cell transplantation (HSCT) all were still relapse-free survivors. Conclusions: Decitabine alone or combination with AAG can improve outcome of AML/MDS with a complex karyotype, there being no significant difference decitabine in inducing remission rates in patients with different karyotype. Increasing the number of courses can improve efficiency. This approach with fewer treatment side effects in patients with a better tolerance should be employed in order to create an improved subsequent chance for HSCT.