1.Comparison of response to LPS-induced sepsis in three DBA/2 stocks derived from different sources
Ji Won PARK ; Su Jin LEE ; Ji Eun KIM ; Mi Ju KANG ; Su Ji BAE ; Yun Ju CHOI ; Jeong Eun GONG ; Kil Soo KIM ; Young-Suk JUNG ; Joon-Yong CHO ; Yeon Shik CHOI ; Dae Youn HWANG ; Hyun Keun SONG
Laboratory Animal Research 2021;37(1):44-52
		                        		
		                        			
		                        			Sepsis, one of the most fatal diseases in the world, is known to culminate in multiple organ failure due to an uncontrolled inflammatory response. Hence, the use of animal models in sepsis research is very important to study complex immune responses. The current study was undertaken to compare commercial stocks with KFDA stocks of DBA/2 mice as an animal model for sepsis study. To compare responses of DBA/2 mice to lipopolysaccharides (LPS)-induced sepsis, we measured altered characteristics of various factors associated with sepsis, including survival curves, organ failure and inflammatory response, in DBA/2Korl stock and two commercial stocks (DBA/2A and DBA/ 2B). Survival rates after LPS exposure were similar for DBA/2Korl and DBA/2B; however, for times over 20 h, survival rates were reduced and concentration dependent in DBA/2A. In order to evaluate multiple organ failure caused by sepsis, H&E stains were evaluated for liver and spleen tissues obtained in the early (2 h) and later (20 h) stages after exposure to LPS; no significant differences were observed between the three stocks. mRNA and protein levels of proinflammatory cytokines were assessed for evaluating inflammatory reactions, and were found to increase in a dose-dependent manner in most DBA/2 mice after LPS treatment. However, no changes were observed in the mRNA levels of proinflammatory cytokines at 20 h after LPS exposure in the DBA/2A stock. The induction of inflammation-mediated factors by LPS exposure did not induce alterations in the mRNA levels of COX-2 and iNOS in all three DBA/2 stocks. Our results indicate that response of DBA/2Korl to LPS-induced sepsis is similar to the two commercial DBA/2 stocks, thus representing its potential as a useful biological resource established in Korea.
		                        		
		                        		
		                        		
		                        	
2.Comparison of response to LPS-induced sepsis in three DBA/2 stocks derived from different sources
Ji Won PARK ; Su Jin LEE ; Ji Eun KIM ; Mi Ju KANG ; Su Ji BAE ; Yun Ju CHOI ; Jeong Eun GONG ; Kil Soo KIM ; Young-Suk JUNG ; Joon-Yong CHO ; Yeon Shik CHOI ; Dae Youn HWANG ; Hyun Keun SONG
Laboratory Animal Research 2021;37(1):44-52
		                        		
		                        			
		                        			Sepsis, one of the most fatal diseases in the world, is known to culminate in multiple organ failure due to an uncontrolled inflammatory response. Hence, the use of animal models in sepsis research is very important to study complex immune responses. The current study was undertaken to compare commercial stocks with KFDA stocks of DBA/2 mice as an animal model for sepsis study. To compare responses of DBA/2 mice to lipopolysaccharides (LPS)-induced sepsis, we measured altered characteristics of various factors associated with sepsis, including survival curves, organ failure and inflammatory response, in DBA/2Korl stock and two commercial stocks (DBA/2A and DBA/ 2B). Survival rates after LPS exposure were similar for DBA/2Korl and DBA/2B; however, for times over 20 h, survival rates were reduced and concentration dependent in DBA/2A. In order to evaluate multiple organ failure caused by sepsis, H&E stains were evaluated for liver and spleen tissues obtained in the early (2 h) and later (20 h) stages after exposure to LPS; no significant differences were observed between the three stocks. mRNA and protein levels of proinflammatory cytokines were assessed for evaluating inflammatory reactions, and were found to increase in a dose-dependent manner in most DBA/2 mice after LPS treatment. However, no changes were observed in the mRNA levels of proinflammatory cytokines at 20 h after LPS exposure in the DBA/2A stock. The induction of inflammation-mediated factors by LPS exposure did not induce alterations in the mRNA levels of COX-2 and iNOS in all three DBA/2 stocks. Our results indicate that response of DBA/2Korl to LPS-induced sepsis is similar to the two commercial DBA/2 stocks, thus representing its potential as a useful biological resource established in Korea.
		                        		
		                        		
		                        		
		                        	
3.The Relevance of the Primary Prevention Criteria for Implantable Cardioverter Defibrillator Implantation in Korean Symptomatic Severe Heart Failure Patients.
JiYeong KIM ; Eue Keun CHOI ; Min Ho LEE ; Do Yoon KANG ; Young Jun SUNG ; Dong Won LEE ; Ilyoung OH ; Yun Shik CHOI ; Seil OH
Korean Circulation Journal 2012;42(3):173-183
		                        		
		                        			
		                        			BACKGROUND AND OBJECTIVES: Implantable cardioverter defibrillator (ICD) therapy is recommended as the primary tool for prevention of sudden cardiac death (SCD) in symptomatic patients with severe left ventricular dysfunction. There is a paucity of information on whether this recommendation is appropriate for the Korean population with severe heart failure. SUBJECTS AND METHODS: The study group consisted of 275 consecutive patients (mean age 65 years, 71% male) who met the ICD implantation criteria for primary prevention (left ventricular ejection fraction < or =30% and New York Heart Association functional class II or III). We analyzed the clinical characteristics and outcomes of an ischemic cardiomyopathy (ICMP) group (n=131) and a non-ischemic cardiomyopathy (NICMP) group (n=144). The outcomes of these 2 groups were compared with the Multicenter Automatic Defibrillator Implantation Trial II (MADIT-II) conventional and Defibrillators in the Non-ischemic Cardiomyopathy Treatment Evaluation (DEFINITE) standard therapy groups, respectively. RESULTS: Eighty patients (29%) died during a follow-up period of 40+/-17 months. The NICMP group had better all-cause mortality rates than the ICMP group (19% vs. 40%, p<0.001), however both groups had a similar incidence of SCD (7% vs. 10%, p=0.272). The 2-year all-cause mortality and SCD for the ICMP group were similar to those of the MADIT-II conventional therapy group (20% vs. 20%, 7% vs. 10%, respectively, all p>0.05). All-cause mortality and the incidence of SCD in the NICMP group were comparable to those of the DEFINITE standard therapy group (13% vs. 17%, 6% vs. 6%, respectively, all p>0.05). CONCLUSION: Korean patients with severe heart failure in both the ICMP and NICMP groups had all-caused mortality and risk of SCD comparable to patients in the MADIT-II and DEFINITE standard therapy groups. Therefore, the primary prevention criteria for ICD implantation would be appropriate in both Korean ICMP and NICMP patients.
		                        		
		                        		
		                        		
		                        			Cardiomyopathies
		                        			;
		                        		
		                        			Death, Sudden, Cardiac
		                        			;
		                        		
		                        			Defibrillators
		                        			;
		                        		
		                        			Defibrillators, Implantable
		                        			;
		                        		
		                        			Follow-Up Studies
		                        			;
		                        		
		                        			Heart
		                        			;
		                        		
		                        			Heart Failure
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Incidence
		                        			;
		                        		
		                        			New York
		                        			;
		                        		
		                        			Primary Prevention
		                        			;
		                        		
		                        			Stroke Volume
		                        			;
		                        		
		                        			Ventricular Dysfunction, Left
		                        			
		                        		
		                        	
4.Assessment of Intermediate Coronary Stenosis in Koreans Using the Fractional Flow Reserve.
Keun Ho PARK ; Bon Kwon KOO ; Jung Won SUH ; Hae Young LEE ; Jin Shik PARK ; Hyun Jae KANG ; Young Seok CHO ; Woo Young CHUNG ; Tae Jin YOUN ; In Ho CHAE ; Dong Ju CHOI ; Hyo Soo KIM ; Byung Hee OH ; Young Bae PARK
Korean Circulation Journal 2008;38(9):468-474
		                        		
		                        			
		                        			BACKGROUND AND OBJECTIVES: Previous studies that used physiologic parameters have shown that coronary angiography is not always accurate in the evaluation of intermediate lesions. We sought to evaluate the outcomes of Korean patients for whom revascularization was deferred according to their fractional flow reserve (FFR). SUBJECTS AND METHODS: FFR was measured in 107 intermediate lesions (visually estimated percent stenosis: 40-70%) in 102 consecutive patients (68% males, mean age: 62+/-9 years). The one-year cardiac adverse outcomes (cardiac death, myocardial infarction or target vessel revascularization) of all the patients and the long-term outcomes of the patients for whom revascularization was deferred according to the FFR were evaluated. RESULTS: The mean percent diameter stenosis was 57+/-11% and the FFR was 0.82+/-0.10. Only 25 lesions (23%) had a FFR <0.75. There was no significant difference in the 1-year cardiac event rates between the FFR > or =0.75 group and the FFR <0.75 group (10.4% vs. 13.0%, respectively, p=0.49). There was a tendency of a lower incidence of 1-year cardiac events in the medical treatment group than in the revascularizaton group (8.0% vs. 20.0%, respectively, p=0.10). In the 69 patients with FFR-guided deferral of revascularization, the long-term (mean follow-up duration: 41+/-11 months) cardiac event-free survival rate was 86%. CONCLUSION: The measurement of FFR seems to be a useful guide for decision making and it may reduce unnecessary intervention in Korean patients who suffer with intermediate stenosis.
		                        		
		                        		
		                        		
		                        			Constriction, Pathologic
		                        			;
		                        		
		                        			Coronary Angiography
		                        			;
		                        		
		                        			Coronary Stenosis
		                        			;
		                        		
		                        			Decision Making
		                        			;
		                        		
		                        			Disease-Free Survival
		                        			;
		                        		
		                        			Follow-Up Studies
		                        			;
		                        		
		                        			Fractional Flow Reserve, Myocardial
		                        			;
		                        		
		                        			Glycosaminoglycans
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Incidence
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Myocardial Infarction
		                        			;
		                        		
		                        			Stents
		                        			
		                        		
		                        	
5.Increased Inflammatory Markers and Endothelial Dysfunction are Associated with Variant Angina.
Sook Hee CHO ; In Hyae PARK ; Myung Ho JEONG ; Seon Ho HWANG ; Nam Shik YUN ; Seo Na HONG ; Sang Rok LEE ; Kye Hun KIM ; Yun MOON ; Young Joon HONG ; Hyung Wook PARK ; Ju Han KIM ; Young Keun AHN ; Jeong Gwan CHO ; Jong Chun PARK ; Jung Chaee KANG
Korean Circulation Journal 2007;37(1):27-32
		                        		
		                        			
		                        			BACKGROUND AND OBJECTIVES: Endothelial dysfunction and increased vascular inflammation may be associated with variant angina (VA). However, their exact roles remain to be clarified. The aim of the presents study is to investigate whether the level of inflammation markers and the flow-mediated dilation (FMD) are related to VA. SUBJECTS AND METHODS: The study included 46 patients (VA group: 53.9+/-12.0 years, 20 males) with positive spasm provocation tests and they were without significant coronary stenosis, and 14 patients (control group: 46.6+/-13.5 years, 7 males) with negative spasm provocation tests and they were without significant coronary stenosis. The clinical characteristics and inflammatory markers, including the high sensitive C-reactive protein (hsCRP) level, the monocyte count and the von Willebrand factor (vWF) level, and the FMD were compared between the two groups. The FMD and inflammatory markers were measured in the morning before performing the ergonovine provocation coronary angiogram. RESULTS: The level of vWF was significantly higher in the VA group than in the control group (166.5+/-41.9% vs. 118.0+/-65.3%, respectively, p=0.029). The FMD was significantly decreased in the VA group compared with the control group (9.2+/-4.3% vs. 12.4+/-4.2%, respectively, p=0.021). Nitrate-mediated dilation did not differ between the two groups. The levels of the monocyte count, hs-CRP and homocysteine were higher in the VA group than in the control group (554.7+/-261.0/mm3 vs. 440.7+/-136.0/mm3, respectively, p=0.039; 0.3+/-0.4 mg/dL vs. 0.1+/-0.1 mg/dL, respectively, p=0.029; 7.54+/-4.0micronmol/L vs. 5.92+/-1.6micronmol/L, respectively, p=0.033). CONCLUSION:  The results of this study suggested that increased inflammatory markers and endothelial dysfunction may be associated with variant angina.
		                        		
		                        		
		                        		
		                        			Angina Pectoris
		                        			;
		                        		
		                        			C-Reactive Protein
		                        			;
		                        		
		                        			Coronary Stenosis
		                        			;
		                        		
		                        			Endothelium
		                        			;
		                        		
		                        			Ergonovine
		                        			;
		                        		
		                        			Homocysteine
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Inflammation
		                        			;
		                        		
		                        			Monocytes
		                        			;
		                        		
		                        			Spasm
		                        			;
		                        		
		                        			von Willebrand Factor
		                        			
		                        		
		                        	
6.Clinical Analysis of Recurrent Chronic Subdural Hematoma.
Hyoung Lae KANG ; Hyung Shik SHIN ; Tae Hong KIM ; Yong Soon HWANG ; Sang Keun PARK
Journal of Korean Neurosurgical Society 2006;40(4):262-266
		                        		
		                        			
		                        			OBJECTIVE: Chronic subdural hematoma(CSDH) is usually treated by burr hole trephination and hematoma evacuation with closed drainage and the surgical result is relatively good in most reported series. But, some patients experience the recurrence of CSDH. We study the clinical factors related to the recurrence of CSDH. METHODS: 213 consecutive patients with CSDH who were treated with burr hole trephination and hematoma evacuation with closed drainage. The medical records, radiologic findings were reviewed retrospectively and clinical factors associated with the recurrent CSDH were analysed statistically. RESULTS: 8.4%(18 cases) of the 213 patients who were treated due to CSDH were recurred. The demographic variables such as age, sex, coexisting diseases were not related to the recurrence. The preoperative hematoma thickness and postoperative hematoma thickness were not associated with the recurrence. The only factor related to the recurrence is postoperative hematoma density in this study. CONCLUSION: This study shows that postoperative hematoma density was strongly related to the recurrence of CSDH. However, several factors associated with the recurrent CSDH were reported in the liletrature. Thus, further study will be needed to uncover the factors related to the recurrence of CSDH. Chronic subdural hematoma(CSDH);Recurrence;Postoperative hematoma density.
		                        		
		                        		
		                        		
		                        			Drainage
		                        			;
		                        		
		                        			Hematoma
		                        			;
		                        		
		                        			Hematoma, Subdural, Chronic*
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Medical Records
		                        			;
		                        		
		                        			Recurrence
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Trephining
		                        			
		                        		
		                        	
7.Intraexaminer Variability of Nerve Conduction Study in Healthy Person.
Seo Hyun KIM ; Seung Min KIM ; Yeon Kyung JUNG ; Sa Yoon KANG ; Soon Hee KWON ; Sook Young ROH ; Keun Ho CHEONG ; Joon Shik MOON ; Ki Han KWON ; Jeong Mo NAM ; Il Nam SUNWOO
Journal of the Korean Neurological Association 2004;22(3):235-239
		                        		
		                        			
		                        			BACKGROUND: Nerve conduction study (NCS) is an objective and quantitative test in evaluating peripheral nerve disorders. Several physiological and technical factors are well known to influence the results of NCS, which can be controlled and regulated by standardization of environment and through the process to make range of normality. However, most electromyographers do not pay much attentions about inter- and intra-examiner variabilities, and there are only a few and incomplete reports on these topics. We examined the intra-examiner variability of NCS on the basis of periods of practice. METHODS: Twenty-eight electromyographers were divided into two groups: residents and neurologist-technicians. All, having variable NCS training periods, have performed NCS on one of other 27 electromyographers ten times within two weeks where each study was made once a day. RESULTS: Coefficient of variation and external quotient increased according to the following order - nerve conduction velocities (NCV), terminal latencies (TL), and amplitudes of compound action potentials (AMP). There were significant differences between the two groups in NCV and TL, but no statistical difference in AMP. CONCLUSIONS: Our results suggest that errors from intra-examiner variability should be considered when interpreting NCS and that those electromyographers who have enough training should perform NCS.
		                        		
		                        		
		                        		
		                        			Action Potentials
		                        			;
		                        		
		                        			Attention
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Neural Conduction*
		                        			;
		                        		
		                        			Peripheral Nerves
		                        			
		                        		
		                        	
8.Endothelium-Dependent Relaxation Is Impaired by Environmental Tobacco Smoke Exposure in Guinea Pig Aorta.
Young Keun ON ; Min Jong KANG ; Seok Yeon KIM ; Hae Young LEE ; In Ho CHAE ; Cheol Ho KIM ; Dae Won SOHN ; Byung Hee OH ; Myoung Mook LEE ; Young Bae PARK ; Yun Shik CHOI
Korean Circulation Journal 2000;30(9):1149-1155
		                        		
		                        			
		                        			BACKGROUND & OBJECTIVES: Smoking is a risk factor for the development of atherosclerosis. However, the exact mechanism of smoking-related damage to the arterial wall and its relation to the atherosclerotic process are not known. The endothelium plays an important role for the regulation of vascular tone and its function is impaired in the presence of risk factors early in the process of atherosclerosis. To assess the effect of smoking on endothelium-dependent relaxation, we examined vascular reactivity in vitro in an animal model of environmental tobacco smoke exposure. MATERIALS & METHODS: Guinea pigs were randomized to control group or environmental tobacco smoke exposure group. The source of environmental tobacco smoke exposure was sidestream smoke during 6 hours per day and 5 days per week over 6 weeks or 12 weeks in a smoking chamber. Fresh aortic rings were suspended in organ baths(37degrees C, 95%O2-5%CO2). Rings were precontracted with phenylephrine(10(-7)M) and exposed to acetylcholine(10(-7)-10(-4)M) and sodium nitroprusside(10(-7)-10(-5)M) in increasing doses and isometric tension was recorded to evaluate the endothelium-dependent and -independent relaxation. RESULTS: Acetylcholine-induced maximal relaxation(mean+/-SD) was 18+/-9% in the rings from the 6-week smoked guinea pigs and 4+/-3% in the rings from the 12-week smoked guinea pigs in comparison with 49+/-6% of the controls(p<0.05). The endothelium-dependent relaxation of the rings from 12 week-exposure was significantly less than that of 6 week-exposure. In contrast endothelium-independent relaxation to SNP was not different among three groups. CONCLUSION: Environmental tobacco smoke exposure impairs endothelial function in guinea pig aorta.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Aorta*
		                        			;
		                        		
		                        			Atherosclerosis
		                        			;
		                        		
		                        			Endothelium
		                        			;
		                        		
		                        			Guinea Pigs*
		                        			;
		                        		
		                        			Guinea*
		                        			;
		                        		
		                        			Models, Animal
		                        			;
		                        		
		                        			Relaxation*
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Smoke*
		                        			;
		                        		
		                        			Smoking
		                        			;
		                        		
		                        			Sodium
		                        			;
		                        		
		                        			Tobacco*
		                        			
		                        		
		                        	
9.Effects of Sophorae Radix Extracts on the Expression of Cell Cycle Regulatory Proteins in Human Gingival Fibroblasts.
Heung Shik KIM ; Hyun A KIM ; Yong Ouk YOU ; Tai Hyun KANG ; Youn Chul KIM ; Tak KIM ; Sung Hee PEE ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 2000;30(4):869-883
		                        		
		                        			
		                        			Fibroblasts are major cellular components of gingiva and periodontal ligament. They regulate the healing process after surgery or injury. Recently, many natural medicines, whose advantages are less side effects and possibility of long-term use, have been studied for their capacity, their anti-bacterial and anti-inflammatory effects and regenerative potential of periodontal tissues. Sophorae radix have been traditionally used as an antibacterial and anti-inflammatory drug in oriental medicine. The purpose of present study was to investigate the effects of Sophorae radix extract on cell cycle progression and its molecular mechanism in human gingival fibroblasts. Sophorae radix extracts(100microgram/ml) notably increased cell proliferation and cell activity in the human gingival fibroblasts as compared to non-supplemented controls. There was an increase in the S phase and a decrease in the G1 phase in 100microgram/ml of Sophorae radix extracts group as compared to non-supplemented controls. The level of cyclin E and cdk 2 protein in test group was higher than that of control groups. But that of cyclin D, cdk 4, and cdk 6 was not distinguished from controls. The level of p53 protein in test group was lower than that of controls, whereas that of p21 was not different. The level of pRB protein in test group was higher than that of controls, whereas that of p16 was lower. These results indicate that the increase of cell proliferation by Sophorae radix extracts may be due to the increased expression of cyclin E and cdk 2, and the decreased expression of p53 and p16 in human gingival fibroblasts.
		                        		
		                        		
		                        		
		                        			Cell Cycle Proteins*
		                        			;
		                        		
		                        			Cell Cycle*
		                        			;
		                        		
		                        			Cell Proliferation
		                        			;
		                        		
		                        			Cyclin D
		                        			;
		                        		
		                        			Cyclin E
		                        			;
		                        		
		                        			Cyclins
		                        			;
		                        		
		                        			Fibroblasts*
		                        			;
		                        		
		                        			G1 Phase
		                        			;
		                        		
		                        			Gingiva
		                        			;
		                        		
		                        			Humans*
		                        			;
		                        		
		                        			Medicine, East Asian Traditional
		                        			;
		                        		
		                        			Periodontal Ligament
		                        			;
		                        		
		                        			S Phase
		                        			;
		                        		
		                        			Sophora*
		                        			
		                        		
		                        	
10.Effect of Extract of Seeds of Carthamus tinctorius L. on Mineralization in Periodontal Ligament Cells and Osteoblastic Cells.
Jung Goo KANG ; Hyung Keun YOU ; Hyung Shik SHIN
The Journal of the Korean Academy of Periodontology 1998;28(3):475-489
		                        		
		                        			
		                        			The main goal for the treatment of periodontal diseases is the regeneration of lost cementum, bone and connective tissue. Clinical and histological research suggests that it is possible to restore periodontal structures. Seeds of Carthamus tinctorius L. has been used for the treatment of bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine the effect of extract of seeds of Carthamus tinctorius L. on mineralization in periodontal ligament cells and osteoblastic cells. Periodontal ligament cells were primarily obtained from a extracted premolars with non-periodontal diseases. Osteoblastic cells were obtained from calvariae of a fetal rat. Cells were cultured with DMEM at 37degrees C with 5% CO2 in 100% humidity incubator. Alkaline phosphatase(ALP) level and the number of calcification nodules were examined and western blot analysis using osteonectin was performed. Measurements of ALP levels and calcification nodules showed that extract of seeds of Carthamus tinctorius L. had significantly higher activity than control in all of both cells. In western blot analysis, protein expression of osteonectin indicated that extract of seeds of Carthamus tinctorius L. showed an increased pattern than control in all of both cells. From the above results, it seems that extract of seeds of Carthamus tinctorius L. has excellent effect on mineralization in periodontal ligament cells and osteoblastic cells.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Bicuspid
		                        			;
		                        		
		                        			Blotting, Western
		                        			;
		                        		
		                        			Carthamus tinctorius*
		                        			;
		                        		
		                        			Carthamus*
		                        			;
		                        		
		                        			Connective Tissue
		                        			;
		                        		
		                        			Dental Cementum
		                        			;
		                        		
		                        			Fractures, Bone
		                        			;
		                        		
		                        			Humidity
		                        			;
		                        		
		                        			Incubators
		                        			;
		                        		
		                        			Osteoblasts*
		                        			;
		                        		
		                        			Osteonectin
		                        			;
		                        		
		                        			Osteoporosis
		                        			;
		                        		
		                        			Periodontal Diseases
		                        			;
		                        		
		                        			Periodontal Ligament*
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Regeneration
		                        			;
		                        		
		                        			Skull
		                        			
		                        		
		                        	
            
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