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The Effect of Insulin-Like Growth Factor-I(IGF-I) and IGF Binding Protein-3(IGFBP-3) on Cellular Proliferation in Mouse 3T3 Fibroblast Cells (마우스 섬유아세포(3T3 fibroblast cells)에서 Insulin-like Growth Factor-I(IGF-I) 및 IGF Binding Protein-3 (IGFBP-3)이 세포증식에 미치는 영향)

  • Cho, Chul-Ho;Kwak, Seung-Min;Moon, Tae-Hun;Cho, Jae-Hwa;Ryu, Jeong-Seon;Lee, Hyong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.5
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    • pp.618-628
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    • 1999
  • Background: Cell growth is a balance between cell proliferation and cell death. Insulin-like growth factor-I(IGF-I), which binds IGF-I receptor(IGF-IR), mediates cellular proliferation as a potent mitogen. IGF binding protein-3(IGFBP-3) as a circulating major IGFBP can inhibit or enhance the effects of IGF-I on cellular growth by binding IGFs. Methods: We investigated the expressions of mRNA of IGF-I and IGF-IR by northern blot and phosphorylation of IGF-IR with the treatment of IGF-I by western blot in 3T3 fibroblast cells. The cellular proliferations of 3T3 cells with the treatments of IGF-I were evaluated using $^3H$-thymidine incorporation and MTT assay. Also to observe the effect of IGFBP-3 on cellular proliferation, 3T3 cells were treated with anti-IGFBP-3 and ${\alpha}IR_3$(monoclonal antibody to IGF-IR) alone or in combination. Results: Our results demonstrated that 3T3 cells showed mRNA expressions of IGF-I and IGF-IR and the IGF-I increased phosphorylation of IGF-IR. The treatments of 3T3 cells with IGF-I increased cellular proliferation in 5 % and 1 % seruma-containing media, not in serum-free media. The addition of anti-IGFBP-3 to neutralize IGFBP-3 showed 2-fold increase of cellular proliferation, and also co-incubation of anti-IGFBP-3 and ${\alpha}IR_3$ together showed similar increase of cellular proliferation in 3T3 cells. Interestingly, when the cells were pretreated with ${\alpha}IR_3$ for 4 hr, prior to the simultaneous addition of ${\alpha}IR_3$ and anti-IGFBP-3, anti-IGFBP-3-mediated cellular proliferation was decreased to control level. All of these results suggest that free IGF-I released from IGF-I/IGFBP-3 complex would be involved in the cellular proliferation. Conclusion: IGF-I is a mitogen through the activation of IGF-IR in 3T3 cells, and IGFBP-3 could be a potent inhibitor for IGF-I action by binding IGF-I.

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Angiotensinogen gene M235T polymorphism as a predictor of cardiovascular risk in hypertensive adolescents (고혈압 청소년의 심혈관계 위험요소로서 Angiotensinogen M235T 유전자 다형)

  • Gil, Joo Hyun;Lee, Jung Ah;Park, Eun Young;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.52 no.1
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    • pp.36-43
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    • 2009
  • urpose : The renin-angiotensin system (RAS) has been demonstrated to play a major role in regulating blood pressure. Therefore, components of the RAS are likely candidate genes that may predispose an individual to essential hypertension and cardiovascular complications. Among them, the M235T polymorphism of the angiotensinogen gene has been speculated to be associated with elevated circulating angiotensinogen concentrations and essential hypertension. This study aimed to analyze the angiotensinogen M235T polymorphism in hypertensive adolescents and investigate its relationship with cardiovascular risks. Methods : Forty Korean hypertensive adolescents (aged 16-17, systolic $BP{\geq}140 mmHg$ and/or diastolic $BP{\geq}90 mmHg$) and fifty seven normal adolescents were included. Obesity index (OI), body mass index (BMI) were calculated. BP was measured by oscillometric methods in resting state. Polymerase chain reaction (PCR) technique was performed on DNA from the hypertensives subjects to analyze the M235T polymorphism. Serum homocysteine, insulin, renin, aldosterone and angiotensin converting enzyme (ACE) were tested according to each genotype. The carotid intima-media thickness (IMT) and carotid artery diameter, Pulse wave velocity (PWV) and ankle-brachial index (ABI) were measured according to each genotype. Results : Genotype frequencies of T/T, M/T and M/M were 62.5%, 35.0%, 2.5%, respectively in hypertensive adolescents. The results were not significantly different compared to control group. Serum insulin, renin levels, BMI and OI were significantly higher in thoses with the M/M genotype as compared to those with the T/T of M/T genotype. Conclusion : This study showed that the M235T polymorphism was not associated with essential hypertension or any cardiovascular risks. Further clinical research is required to ascertain the relationship between this polymorphism and cardiovascular complications in Korean hypertensive adolescents.

Developmental Genetic Analysis of Avian Primordial Germ Cells and the Application to Poultry Biotechnology

  • Kagami, H.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.135-142
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    • 2001
  • A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

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Effects of Applying Cattle Manure on Carrying Capacity of Organic Livestock per Unit Area of Summer Forage Crops (우분뇨 시용이 하계사료작물의 단위면적당 유기가축 사육능력에 미치는 영향)

  • Jo, Ik-Hwan
    • Korean Journal of Organic Agriculture
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    • v.19 no.2
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    • pp.185-198
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    • 2011
  • This study was carried out to select a proper forage crop, and to estimate the proper level of application of cattle manure and carrying capacity of organic livestock per unit area. Corns and forage sorghum hybrids were cultivated with different types of livestock manures and different amount of them to produce organic forage. For both corns and forage sorghum hybrids, no fertilizer plots had significantly (p<0.05) lower annual dry matter (DM), crude protein (CP) and total digestible nutrients (TDN) yields than those of other plots, whereas the N-P-K (nitrogen-phosphorous-kalium) plots ranked the highest yields, followed by 150% cattle manure plots and 100% cattle manure plots. DM, CP and TDN yields of in cattle manure plots were significantly (p<0.05) higher than those of no fertilizer and P-K (phosphorous-kalium) plots. The yields of in cattle slurry plots tended to be a little higher than those of in composted cattle manure plots. Assuming that corn and forage sorghum hybrids produced from this trial were fed at 70% level to 450kg of Hanwoo heifer for 400g of average daily gain, the carrying capacity (head/year/ha) of livestock ranked the highest in 150% cattle slurry plots (mean 6.0 heads), followed by 100% cattle slurry plots (mean 5.3 heads), 150% composted cattle manure plots (mean 4.7 heads), 100% composted cattle manure plots (mean 4.4 heads), and no fertilizer plots (mean 2.8 heads) in corns (or the cultivation of corns). Meanwhile, in the case of forage sorghum hybrids, 150% cattle slurry plots (mean 6.4 heads) ranked the highest carrying capacity, followed by 150% composted cattle manure plots (mean 4.8 heads), 100% cattle slurry plots (mean 4.4 heads), 100% composted cattle manure plots (mean 4.1 heads), and no fertilizer plots (mean 2.8 heads). The results indicated that the application of livestock manure to cultivated soil could enhance not only DM and TDN yields, but also the carrying capacity of organic livestock as compared with the effect of chemical fertilizers. In conclusion, the production of organic forage with reutilized livestock manure will facilitate the reduction of environmental pollution and the production of environmentally friendly agricultural products by resource circulating system.

In Vitro Stability of Liposomes Containing Newly Synthesized Glycolipid (신규 합성 당지질 함유 리포솜의 In Vitro 안정성)

  • Song, Chung-Kil;Jung, Soon-Hwa;Seong, Ha-Soo;Cho, Sun-Hang;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.51 no.1
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    • pp.43-50
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    • 2007
  • Liposomes having particle size from several tens to hundreds nanometers are efficient carriers for injectable drug delivery. Enhancement of liposome stability in bloodstream has been studied because of its relatively short circulation time and fast clearance from human body by reticuloendothelial system (RES) in blood vessel. In this study, new disaccharide-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) derivatives in which lactose or sucrose as the disaccharide molecule was conjugated covalently to DSPE were synthesized. Liposomes of which surface had disaccharide molecules were prepared by incorporating the disaccharide-DSPE into liposomes as one of their lipid components. Particle size of the prepared liposomes was approximately 100 nm. The liposomes of which surface were modified with the disaccharide-DSPE showed -25 mV of zeta potential value due to the presence of hydroxyl groups on their surface, while the unmodified control liposomes showed -10 mV of zeta potential value. Loading efficiency of model drug, doxorubicin, into liposomes was about 90%. Stability of the disaccharide-modified liposomes in vitro was evaluated by monitoring the amount of protein adsorption and particle size of the liposomes in serum. Disaccharide-modified liposomes were more stable in serum than unmodified control liposomes or polyethyleneglycol (PEG)-modified liposomes due to less adsorption of serum protein and hence less increase of their particle size. The liposomes of which surface was modified with disaccharide-DSPE conjugate can be used as long-circulating carriers for drugs having high toxicity or short half-life time due to their enhanced stability in blood circulatory system.

The inhibitory effect of physiological pattern of testosterone on luteinizing hormone secretion in sheep (면양에서 생리적 분비형태의 테스토스테론이 황체형성호르몬의 분비 억제에 미치는 효과)

  • Rhim, Tae-jin;Park, Kyoung-sik
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.271-278
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    • 1995
  • The objective of the present study was to investigate the inhibitory effect of physiologically pulsatile pattern of testosterone(T) on luteinizing hormone(LH) in wethers. To do this, 3 separate experiments were conducted. Infusion rates and patterns needed to produce normal T secretory profiles found in intact rams were established in Experiment 1, the time-course of the suppressive effect of T on circulating LH concentrations was determined in Experiment 2, and the effectiveness of a pulsatile versus a constant pattern of T to suppress LH secretion in wethers was compared in Experiment 3. In Experiment 1, three different doses(25, 50 or $100{\mu}g$) of T were injected intravenously to animals to do pharmacokinetic analysis of T. Elimination rate constant, volume of distribution, and total body clearance of T averaged $0.18min^{-1}$, 0.531/kg BW, and 0.091/min/ kg BW, respectively. In Experiment 2, three different doses(192,384, or $768{\mu}g/kg/24h$) of T were infused at 4h intervals for 3 days into animals to evaluate the time course of the inhibitory effect of T on mean LH concentration. As duration of T infusion increased, mean LH concentrations gradually reduced. Mean LH concentrations were significantly lower at day 2 or day 3 than at day 0. However, mean LH concentrations did not differ between day 0 and day 1 or between day 2 and day 3. In Experiment 3, animals were subjected to two different intravenous infusion regimens for 3 days: constant T($768{\mu}g/kg/24h$) and pulsatile(one pulse every 4h) T($768{\mu}g/kg24h$). Blood samples were collected at 10-min intervals for 4h both prior to infusion and during the last 4h of the infusion. Mean LH was more suppressed(p=0.045) by constant T than by pulsatile T. LH pulse amplitude was not affected by constant T or pulsatile T. LH interpulse interval was increased more(p=0.034) by constant T than pulsatile T.

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Rectal Temperature Maintenance Using a Heat Exchanger of Cardioplegic System in Cardiopulmonary Bypass Model for Rats (쥐 심폐바이패스 모델에서 심정지액 주입용 열교환기를 이용한 직장체온 유지)

  • Choi Se-Hoon;Kim Hwa-Ryong;Paik In-Hyuck;Moon Hyun-Jong;Kim Won-Gon
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.505-510
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    • 2006
  • Background: Small animal cardiopulmonary bypass (CPB) model would be a valuable tool for investigating path-ophysiological and therapeutic strategies on bypass. The main advantages of a small animal model include the reduced cost and time, and the fact that it does not require a full scale operating environment. However the rat CPB models have a number of technical limitations. Effective maintenance and control of core temperature by a heat exchanger is among them. The purpose of this study is to confirm the effect of rectal temperature maintenance using a heat exchanger of cardioplegia system in cardiopulmonary bypass model for rats. Material and Method: The miniature circuit consisted of a reservoir, heat exchanger, membrane oxygenator, roller pump, and static priming volume was 40 cc, Ten male Sprague-Dawley rats (mean weight 530 gram) were divided into two groups, and heat exchanger (HE) group was subjected to CPB with HE from a cardioplegia system, and control group was subjected to CPB with warm water circulating around the reservoir. Partial CPB was conducted at a flow rate of 40 mg/kg/min for 20 min after venous cannulation (via the internal juglar vein) and arterial cannulation (via the femoral artery). Rectal temperature were measured after anesthetic induction, a ter cannulation, 5, 10, 15, 20 min after CPB. Arterial blood gas with hematocrit was also analysed, 5 and 15 min after CPB. Result: Rectal temperature change differed between the two groups (p<0.01). The temperatures of HE group were well maintained during CPB, whereas control group was under progressive hypothermia, Rectal temperature 20 min after CPB was $36.16{\pm}0.32^{\circ}C$ in the HE group and $34.22{\pm}0.36^{\circ}C$ in the control group. Conclusion: We confirmed the effect of rectal temperature maintenance using a heat exchanger of cardioplegia system in cardiopulmonary bypass model for rats. This model would be a valuable tool for further use in hypothermic CPB experiment in rats.

Can Peritoneal Dialysis Remove Endothelin-1 after Cardiopulmonary Bypass for Repair of Congenital Heart Disease? (체외순환을 이용한 선천성 심장수술 후에 시행한 복막투석이 내피의존성 혈관 활성 물질인 Endothelin-1을 제거할 수 있는가?)

  • Chang, Yun-Hee;Ban, Ji-Eun;Lee, Hyoung-Doo;Lee, Sun-Hee;Rhym, Byuong-Yong;Sung, Si-Chan
    • Journal of Chest Surgery
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    • v.40 no.4 s.273
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    • pp.247-255
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    • 2007
  • Background: Lung injury that follows bypass has been well described. It is manifested as reduced oxygenation and lung compliance and, most importantly, increased pulmonary vascular resistance reactivity; this is a known cause of morbidity and mortality after repair of congenital heart disease. Injury to the pulmonary vascular endothelium, and its associated alterations of endothelin-1, is considered to be a major factor of bypass-induced lung injury. Removing endothelin-1 after bypass may attenuate this response. This study measured the concentration of serum and peritoneal effluent endothelin-1 after performing bypass to determine if endothelin-1 can be removed via peritoneal dialysis. Material and Method: From March 2005 to March 2006, 18 patients were enrolled in this study Peritoneal catheters were placed at the end of surgery. Serum samples were obtained before and after bypass, and peritoneal effluents were obtained after bypass. Endothelin-1 was measured by enzyme linked immunosorbent assay (ELISA). Result: In the patients with a severe increase of the pulmonary artery pressure or flow, the mean preoperative plasma endothelin-1 concentration was significantly higher than that in the patients who were without an increase of their pulmonary artery pressure or flow (4.2 vs 1.8 pg/mL, respectively, p<0.001). The mean concentration of plasma endothelin-1 increased from a preoperative value of $3.61{\pm}2.17\;to\;5.33{\pm}3.72 pg/ml$ immediately after bypass. After peritoneal dialysis, the mean plasma endothelin-1 concentration started to decrease. Its concentration at 18 hours after bypass was significantly lower than the value obtained immediately after bypass (p=0.036). Conclusion: Our data showed that the plasma endothelin-1 concentration became persistently decreased after starting peritoneal dialysis, and this suggests that peritoneal dialysis can remove the circulating plasma endothelin-1.

Changes of Blood Gases, Plasma Catecholamine Concentrations and Hemodynamic Data in Anesthetized Dogs during Graded Hypoxia Induced by Nitrous Oxide (아산화질소에 의한 점진적 저산소가스 흡입이 혈중 가스치와 Catecholamine치 및 혈역학에 미치는 영향)

  • Kim, Sae-Yeon;Song, Sun-Ok;Bae, Jung-In;Cheun, Jae-Kyu;Bae, Jae-Hoon
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.97-113
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    • 1998
  • The sympathoadrenal system plays an important role in homeostasis in widely varing external environments. Conflicting findings, however, have been reported on its response to hypoxia. We investigated the effect of hypoxia on the sympathoadrenal system in dogs under halothane anesthesia by measuring levels of circulating catecholamines in response to graded hypoxia. Ten healthy mongreal dogs were mechanically ventilated with different hypoxic gas mixtures. Graded hypoxia and reoxygenation were induced by progressively decreasing the oxygen fraction in the inhalation gas mixture from 21%(control) to 15%, 10% and 5% at every 5 minutes, and then reoxygenated with 60% oxygen. Mean arterial pressure, central venous pressure and mean pulmonary arterial pressure were measured directly using pressure transducers. Cardiac output was measured by the thermodilutional method. For analysis of blood gas, saturation and content, arterial and mixed venous blood were sampled via the femoral and pulmonary artery at the end of each hypoxic condition. The concentration of plasma catecholamines was determined by radioenzymatic assay. According to the exposure of graded hypoxia, not only did arterial and mixed venous oxygen tension decreased markedly at 10% and 5% oxygen, but also arterial and mixed venous oxygen saturation decreased significantly. An increased trend of the oxygen extraction ratio was seen during graded hypoxia. Cardiac output, mean arterial pressure and systemic vascular resistance were unchanged or increased slightly. Pulmonary arterial pressure(PAP) and pulmonary vascular resistance(PVR) were increased by 55%, 76% in 10% oxygen and by 82%, 95% in 5% oxygen, respectively(p<0.01). The concentrations of plasma norepinephrine, epinephrine and dopamine increased by 75%, 29%, 24% in 15% oxygen and by 382%, 350%, 49% in 5% oxygen. These data suggest that the sympathetic nervous system was activated to maintain homeostasis by modifying blood flow distribution to improve oxygen delivery to tissues by hypoxia, but hemodynamic changes might be blunted by high concentration of nitrous oxide except PAP and PVR. It would be suggested that hemodynamic changes might not be sensitive index during hypoxia induced by high concentration of nitrous oxide exposure.

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Neutrophil Count and the Inflammation-based Glasgow Prognostic Score Predict Survival in Patients with Advanced Gastric Cancer Receiving First-line Chemotherapy

  • Li, Qing-Qing;Lu, Zhi-Hao;Yang, Li;Lu, Ming;Zhang, Xiao-Tian;Li, Jian;Zhou, Jun;Wang, Xi-Cheng;Gong, Ji-Fang;Gao, Jing;Li, Jie;Li, Yan;Shen, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.945-950
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    • 2014
  • Purpose: To explore the value of systemic inflammatory markers as independent prognostic factors and the extent these markers improve prognostic classification for patients with inoperable advanced or metastatic gastric cancer (GC) receiving palliative chemotherapy. Methods: We studied the prognostic value of systemic inflammatory factors such as circulating white blood cell count and its components as well as that combined to form inflammation-based prognostic scores (Glasgow Prognostic Score (GPS), Neutrophil-Lymphocyte Ratio (NLR), Platelet Lymphocyte Ratio (PLR), Prognostic Index (PI) and Prognostic Nutritional Index (PNI)) in 384 patients with inoperable advanced or metastatic gastric cancer (GC) receiving first-line chemotherapy. Univariate and multivariate analyses were performed to examine the impact of inflammatory markers on overall survival (OS). Results: Univariate analysis revealed that an elevated white blood cell, neutrophil and/or platelet count, a decreased lymphocyte count, a low serum albumin concentration, and high CRP concentration, as well as elevated NLR/PLR, GPS, PI, PNI were significant predictors of shorter OS. Multivariate analysis demonstrated that only elevated neutrophil count (HR 3.696, p=0.003) and higher GPS (HR 1.621, p=0.01) were independent predictors of poor OS. Conclusion: This study demonstrated elevated pretreatment neutrophil count and high GPS to be independent predictors of shorter OS in inoperable advanced or metastatic GC patients treated with first-line chemotherapy. Upon validation of these data in independent studies, stratification of patients using these markers in future clinical trials is recommended.