• Journal of Chest Surgery
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• v.2 no.1
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• pp.85-104
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• 1969

This experiment was carried out to study the effect of rapid hemorrhage on cardiopulmonary hemodynamics of the cooled dogs. Hypothermia was induced by means of body surface cooling with ice water. Lowest esophageal temperatures ranged from 24 to 26 degree. Dogs were bled via the femoral artery into a reservoir in amount of the equivalent blood volume of 3% of body weight of the dogs. Some dogs were reinfused with the same amount of blood which they lost and others infused with 5% dextrose solution. Fourty adult mongrel dogs were divided into three groups: group I[15 dogs]; dogs were bled in normothermic state. Five dogs had no further treatment, but five dogs were reinfused with blood and five infused with 5% dextrose solution 30 minutes after bleeding. GroupII[10 dogs]; dogs were bled as group I after having been cooled. Five dogs were reinfused with blood as group I. Group III[15 dogs]; dogs were first bled and then cooled. Reinfusion procedures were the same as in group l Results were as follow: 1. The heart rate showed a slight decrease after bleeding in group I and then increased over the control level after 60 minutes. After reinfusion and infusion, the heart rate was also increased gradually and after three hours almost returned to the control level. In group II and groupIll, the heart rate decreased remarkably and after reinfusion showed a light increase but after infusion tended to decrease cotinually. 2. The stroke volume showed remarkable decrease after bleeding in group I., and recovered to control level after reinfusion and infusion,and then gradually decreased again. In group III, the stroke volume showed no remarkable change after hypothermia, and tended to decrease after reinfusion. In group III, the stroke volume decreased remarkably after bleeding and hypothermia,and clearly increased after reinfusion and infusion and then returned to control level. 3. Femoral mean pressure declined very rapidly and significantly right after bleeding and showed a remarkable prompt rise after reinfusion and infusion in group I [67% recovery]. On the other hand, it declined remarkably after hypothermia and bleeding and showed a slight rise after reinfusion and infusion in group II[46% recovery] and III [41% recovery]. 4. Venous pressure declined slightly after bleeding and tended to return to the control level after reinfusion and infusion,in group I. In group II, it did not change significantly during hypothermia but showed a slight decline after bleeding and returned toward control level after reinfusion. In group III, it declined slightly after bleeding and showed no significant change after hypothermia and rose over the control level after reinfusion and infusion. 5. Right ventricular systolic pressure decreased markedly after bleeding and then increased progressively after 30 minutes. It increased after reinfusion and infusion as well, approaching the control level in group I. In group II, it showed no significant change during hypothermia, but decreased remarkably after bleeding and then returned to near control level after reinfusion. In group III, it was decreased markedly after bleeding but did not change significantly during hypothermia and showed a slight increase after reinfusion. 6. The respiratory rate increased gradually after bleeding and decreased gradually after reinfusion but did not return to the control level, whereas it decreased near to the control level after infusion,and tended to increase in group I. In group II, it decreased significantly after hypothermia and bleeding but returned near to the control level after reinfusion. In group III, it showed a remarkable decrease after hypothermia and increased slightly after reinfusion and infusion but did not returned to the control level. In group I, the tidal volume decreased slightly after hemorrhage, and increased gradually to near the control level after 3 hours following reinfusion.

• Journal of the Korean Society of Radiology
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• v.85 no.1
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• pp.138-146
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• 2024

Purpose To evaluate whether the image quality of chest radiographs obtained using a camera-type portable X-ray device is appropriate for clinical practice by comparing them with traditional mobile digital X-ray devices. Materials and Methods Eighty-six patients who visited our emergency department and underwent endotracheal intubation, central venous catheterization, or nasogastric tube insertion were included in the study. Two radiologists scored images captured with traditional mobile devices before insertion and those captured with camera-type devices after insertion. Identification of the inserted instruments was evaluated on a 5-point scale, and the overall image quality was evaluated on a total of 20 points scale. Results The identification score of the instruments was 4.67 ± 0.71. The overall image quality score was 19.70 ± 0.72 and 15.02 ± 3.31 (p < 0.001) for the mobile and camera-type devices, respectively. The scores of the camera-type device were significantly lower than those of the mobile device in terms of the detailed items of respiratory motion artifacts, trachea and bronchus, pulmonary vessels, posterior cardiac blood vessels, thoracic intervertebral disc space, subdiaphragmatic vessels, and diaphragm (p = 0.013 for the item of diaphragm, p < 0.001 for the other detailed items). Conclusion Although caution is required for general diagnostic purposes as image quality degrades, a camera-type device can be used to evaluate the inserted instruments in chest radiographs.

• Journal of Chest Surgery
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• v.38 no.8 s.253
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• pp.538-544
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• 2005

Background: Pulmonary artery banding (PAB) is an initial palliative procedure for a diverse group of patients with congenital cardiac anomalies and unrestricted pulmonary blood flow. We proved the usefulness of PAB through retrospective investigation of the surgical indication and risk analysis retrospectively. Material and Method: One hundred and fifty four consecutive patients (99 males and 55 females) who underwent PAB between January 1986 and December 2003 were included. We analysed the risk factors for early mortality and actuarial survival rate. Mean age was $2.5\pm12.8\;(0.2\sim92.7)$ months and mean weight was $4.5\pm2.7\;(0.9\sim18.0)\;kg$. Preoperative diagnosis included functional single ventricle $(88,\;57.1\%)$, double outlet right ventricle $(22,\;14.2\%)$, transposition of the great arteries $(26,\;16.8\%)$, and atrioventricular septal defect $(11,\;7.1\%)$. Coarctation of the aorta or interrupted aortic arch $(32,\;20.7\%)$, subaortic stenosis $(13,\;8.4\%)$ and total anomalous pulmonary venous connection $(13,\;8.4\%)$ were associated. Result: The overall early mortality was $22.1\%\;(34\;of\;154)$, The recent series from 1996 include patients with lower age $(3.8\pm15.9\;vs.\;1.5\pm12.7,\;p=0.04)$ and lower body weight $(4.8\pm3.1\;vs.\;4.0\pm2.7,\;p=0.02)$. The early mortality was lower in the recent group $(17.5\%;\;16/75)$ than the earlier group $(28.5\%;\;18/45)$. Aortic arch anomaly (p=0.004), subaortic stenosis (p=0.004), operation for subaortic stenosis (p=0.007), and cardiopulmonary bypass (p=0.007) were proven to be risk factors for early death in univariate analysis, while time of surgery (<1996) (p=0.026) was the only significant risk factor in multivariate analysis. The mean time interval from PAB to the second-stage operation was $12.8\pm10.9$ months. Among 96 patients who survived PAB, 40 patients completed Fontan operation, 21 patients underwent bidirectional cavopulmonary shunt, and 35 patients underwent biventricular repair including 25 arterial switch operations. Median follow-up was $40.1\pm48.9$ months. Overall survival rates at 1 year, 5 years and 10 years were $81.2\%\;65.0\%,\;and\;63.5\%$ respectively. Conclusion: Although it improved in recent series, early mortality was still high despite the advances in perioperative management. As for conventional indications, early primary repair may be more beneficial. However, PA banding still has a role in the initial palliative step in selective groups.

• Journal of Chest Surgery
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• v.30 no.5
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• pp.471-478
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• 1997

Introduction: The use of rabbits as a cardiopulmonary bypass(CPB) animal model is extremely dif%cult mainly due to technical problems. On the other hand, deep hypothermic circulatory arrest(CA) is used to facilitate surgical repair in a variety of cardiac diseases. Although steroids are generally known to be effective in the treatment of cerebral edema, the protective effects of steroids on the brain during CA are not conclusively established. Objectives of this study are twofold: the establishment of CPB technique in rabbits and the evaluation of preventive effect of steroid on the development of brain edema during CA. Material '||'&'||' Methods: Fifteen New Zealan white rabbits(average body weight 3.5kg) were divided into three experimental groups; control CA group(n=5), CA with Trendelenberg position group(n=5), and CA with Trendelenberg position + steroid(methylprednisolone 30 mglkg) administration group(n=5). After anesthetic induction and tracheostomy, a median sternotomy was performed. An aortic cannula(3.3mm) and a venous ncannula(14 Fr) were inserted, respectively in the ascending aorta and the right atrium. The CPB circuit consisted of a roller pump and a bubble oxygenator. Priming volume of the circuit was approximately 450m1 with 120" 150ml of blood. CPB was initiated at a flow rate of 80~85ml/kg/min, Ten min after the start of CPB, CA was established with duration of 40min at $20^{\circ}C$ of rectal temperature. After CA, CPB was restarted with 20min period of rewarming. Ten min after weaning, the animal was sacrif;cod. One-to-2g portions of the following tissues were rapidly d:ssected and water contents were examined and compared among gr ups: brain, cervical spinal cord, kidney, duodenum, lung, heart, liver, spleen, pancreas. stomach. Statistical significances were analyzed by Kruskal-Wallis nonparametric test. Results: CPB with CA was successfully performed in all cases. Flow rate of 60-100 mlfkgfmin was able to be maintained throughout CPB. During CPB, no significant metabolic acidosis was detected and aortic pressure ranged between 35-55 mmHg. After weaning from CPB, all hearts resumed normal beating spontaneously. There were no statistically significant differences in the water contents of tissues including brain among the three experimental groups. Conclusion: These results indicate (1) CPB can be reliably administered in rabbits if proper technique is used, (2) the effect of steroid on the protection of brain edema related to Trendelenburg position during CA is not established within the scope of this experiment.

• Journal of Preventive Medicine and Public Health
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• v.32 no.2
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• pp.123-129
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• 1999

Objectives: This study was performed to investigate sweets of genetic polymorphisms of glutathione S-transferase M1 (GSTM1), glutathione S-transferase M1 (GSTT1), cytochrome P450 1A1 (CYP1A1) and cytoehrome P450 2E1 (CYP2E1) on lung cancer development. Methods: Ninety-eight lung cancer patients and 98 age-sex matched non-cancer patients hospitalized in Chungbuk National University Hospital form March 1997 to August 1998, were the subjects of this case-control study. Direct interview was done and genotypes of GSTM1, GSTT1, CYP1A1 and CYP2E1 were investigated using multiplex PCR or PCR-RFLP methods with DNA extracted from venous blood. Effects of the polymorphisms of GSTM1, GSTT1, CYP1A1 and CYP2E1, lifestyle factors including smoking, and their interactions on lung rancor were statistically analyzed. Results: GSTM1 was deleted in 67.01% of the cases and 58.16% of the controls, and the odds ratio(95% CI) was 1.46(0.82-2.62). GSTT1 deletion was 58.76% for the lung cancer patients and 50.00% for the controls[OR:1.43(0.81-2.51)]. The frequencies of lle/lle, lle/Val and Val/Val of the CYP1A1 polymorphisms were 59.18-18%, 35.71%, and 5.10% for the cases, and 52.04%, 45.92%, 2.04% for the controls, respectively. Risk of lung cancer was not associated with polymorphism of CYP1A1 ($x^2trend=0.253$, p-value>0.05). The respective frequency of c1/c1 c1/c2, c2/c2 genotypes for CYP2E1 were 50.00%, 42.86%, 7.14% for the lung cancer patients, and 66.33%, 30.61%, 3.06% for the controls $(x^2trend=5.783,\;p<0.05)$. c2 allele was a significant risk factor for lung cancer. We also observed a significant association of cigarette smoking history with lung cancer risk. The odds ratio(95% Cl) of cigarette smoking was 3.03(1.58-5.81). In multiple logistic analysis including genotypes of GSTM1, GSTT1, CYP1A1 and CYP2E1, and smoking habit, only snaking habit came out to be a significant risk factor for lung cancer. Conclusion: Genetic polymorphisms of GSTM1, GSTT1, CYP1A1 and CYP2E1 are not so strongly associated with lung cancer as lifestyle factors including cigarette smoking.

• Tuberculosis and Respiratory Diseases
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• v.40 no.3
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• pp.213-222
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• 1993

Background : Bronchial asthma has been known as an inflmmatory disease. There have been many evidences that polymorphonuclear leukocytes (PMN) might play an important role in the pathogrnesis of asthma. Although many investigators suggested that pneumocyte injury by PMN-derived oxygen radicals may contribute to the pathogenesis of asthma, there has been few report for a direct evidence of oxygen radicals-mediated pneumocyte injury in bronchial asthma. Furthermore the exact mechanism of oxygen radicals-mediated pneumocyte injury is still controversy. This study was designed to establish a direct in vitro evidence and its clinical significance of pneumocyte injury by PMN-derived superoxide anion in bronchial asthma and to elucidate the main mechanism of superoxide anion-mediated pneumocyte injury. Methods : 12 stable asthmatics and 5 healthy volunteers were participated in this study. PMN was separated from peripheral venous blood samples by using dextran sedimentation and Ficoll-Hypaque density gradient separation method. Superoxide anion productions by PMN and plasma SOD activities were measured by spectrophotometric assay using the principle of SOD inhibitable cytochrome c reduction. PMN-mediated pneumocyte injuries were measured by $^{51}Cr$-release assay using A549 pneumocytes and were expressed as percent lysis and percent detachment. Results: 1) PMN from asthmatics produced more amount of superoxide anion compared to PMN from normal subjects ($6.65{\pm}0.58$ vs $2.81{\pm}0.95\;nmol/1{\times}10^6$ cells, p<0.05), and showed an inverse correlation with $FEV_1$(R=-0.63, p<0.05), but no correlation with $PC_{20}$ histamine in asthmatics. 2) Plasma SOD activities were decreased in asthmatics compared to normal subjects but not significant, and showed a positive correlation with $FEV_1$(R=0.63, p<0.05) but no correlation with $PC_{20}$ histamine in asthmatics. 3) There were a positive correlation between plasma SOD activity and superoxide anion production by PMN in normal subjects (R=0.88, p<0.05) but not in asthmatics. 4) PMN-mediated pneumocyte injury was predominantly expressed as cell detachment rather than cell lysis in both groups, and PMN from asthmatics showed more potent cytotoxic effect on A549 pneumocytes compated to PMN from normal subjects. PMN-mediated detachment rather than lysis of A549 pneumocytes was significantly inhibited by in vitro SOD but not by diluted serum. 5) PMN-mediated detachment rather than lysis of A549 pneumocytes showed a good correlation with superoxide anion production by PMN (R=0.90 in normal subjects, R=0.82 in asthmatics, p<0.05) but no correlation with plasma SOD activity. PMN-mediated pneumocyte injuries were not correlated with $FEV_1$ or $PC_{20}$ histamine in asthmatics. 6) There were no significant differences in PMN-mediated pneumocyte injuries between allergic and nonallergic asthmatics. Conclusion : Our results suggest that pneumocyte injury by PMN-derived superoxide anion may partially contribute to the pathogenesis of asthma and that cell detachment rather than cell lysis may be the mechanism of superoxide anion-mediated pneumocyte injury.

• Journal of Preventive Medicine and Public Health
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• v.25 no.2 s.38
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• pp.127-140
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• 1992

This study was done to identify the association between serum copper and zinc levels and the cirrhosis and hepatocellular carcinoma(HCC), and to evaluate its diagnostic value on liver diseases. Sixty-three healthy persons, 60 patients with cirrhosis and 33 patients with hepatocellular carcinoma were rendomly selected and investigated for their general characteristics from October 1990 to August 1991. For analysis of the biochemical markers in liver function test and the serum copper and zinc levels, their fasting venous blood were sampled at 9:00 to 11:00 in the morning and centrifuged to separate the serum within one hour. All the samples were immediately analysed for biochemical markers and stored at $-20^{\circ}C$ in polypropylene tubes further copper and zinc analysis. Mean of serum coppper levels was $91.97{\pm}4.76{\mu}g/dl$ in control, $106.21{\pm}2.73{\mu}g/dl$ in cirrhosis and $127.05{\pm}0.77{\mu}g/dl$ in HCC. The value of HCC was statistically significantly higher than that of the control and cirrhosis(p<0.05). Serum zinc levels were $110.82{\pm}7.24{\mu}g/dl$ in control, $68.10{\pm}5.43{\mu}g/dl$ in cirrhosis and $63.78{\pm}2.20{\mu}g/dl$ in HCC. The values of cirrhosis and HCC were statistically significantly lower than that of control(p<0.05). The Cu/Zn ratio was statiatically significantly different among three groups(p<0.05). Test total protein, albumin, ALP and total bilirubin of biochemical markers of liver function were statistically significantly different among three groups(p<0.05). Differences between cirrhosis and HCC for ALT and AST, and between the control and HCC for direct bilirubin were not statistically significant. Biochemical markers statistically significantly correlated with serum copper and zinc levels and Cu/Zn ratio(p<0.05), were variable in three groups. In multiple logistic regression, odds ratio of serum copper level and Cu/Zn ratio had no statistical significance on the cirrhosis and the HCC, but that of serum sinc was statistically significant as 0.951 and 0.952(p<0.05). Serum copper and zinc levels and Cu/Zn ratio were not statistically significantly different between the cirrhosis and HCC. H\Albumin, ALP, zinc, total bilirubin and age among all variables were selected as main variables for three-group discriminant analysis. Percentage of 'grouped' cases correctly classified by these five variables was 98.4 for control, 73.4 for cirrhosis, 75.7 for HCC and 84.0 for all subjects. This study suggests that zinc level is considered to play a role as diagnostic marker on the hepatic disorders and be more useful than serum copper level and Cu/Zn ratio in diagnosis of the liver diseases.

• Journal of Yeungnam Medical Science
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• v.10 no.2
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• pp.451-474
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• 1993

Lidocaline if frequently administered as a component of an anesthetic : for local or regional nerve blocks, to mitigate the autonomic response to laryngoscopy and tracheal intubation, to suppress the cough reflex, and for antiarrythmic therapy. Diazepam dectease the potential central nervous system (CNS) toxicity of local anesthetic agents but may modify the sitmulant action of lidocaine in addition to their own cardiovascular depressant. The potential cardiovascular toxicity of local anesthetics may be enhanced by the concomitant administration of diazepam. This study was designed to investigate the effects of lidocaine dose and pretreated diazepam to cardiovascular system and plasma concentration of lidocaine. Lidocaine in 100 mcg/kg/min, 200 mcg/kg/min, and 300 mcg/kg/min was given by sequential infusion to dogs anesthetized with halothane-nitrous oxide (Group I). And in group II, after diazepam pretreatment, lidocaine was infused by same way when lidocaine was administered in 100 mcg/kg/min, the low plasma levels ($3.97{\pm}0.22-4.48{\pm}0.36$ mcg/ml) caused a little reduction in cardiovascular hemodynamics. As administered in 200 mcg/kg/min, 300 mcg/kg/min, the higher plasma levels ($7.50{\pm}0.66-11.83{\pm}0.59$ mcg/ml) reduced mean arterial pressure (MAP), cardiac index (CI), stroke index (SI), left ventricular stroke work index (LVSWI), and right ventricular stroke work index (PVSWI) and increased pulmonary artery wedge pressure (PAWP), central venous pressure (CVP), systemic vascular resistance index (SVRI), but was associated with little changes of heart rate (HR), mean pulmonary artery pressure (MPAP), and pulmonary vascular resistance index (PVRI). When lidocaine with pretreated diazepam was administered in 100 mcg/kg/min, the low plasma level, the lower level than when only lidocaine administered, reduced MAP, but was not changed other cardiovascular hemodynamics. While lidocaine was infused in 200 mcg/kg/min, 300 mcg/kg/min in dogs pretreated diazepam, the higher plasma level ($7.64{\pm}0.79-13.79{\pm}0.82$ mcg/ml) was maintained and was associated with reduced CI, SI, LVSWI and incresed PAWP, CVP, SVRI but was a little changes of HR, MPAP, PVRI. After $CaCl_2$ administeration, CI, SI, SVRI, LVSWI was recovered but PAWP, CVP was rather increased than recovered. The foregoing results demonstrate that pretreated diazepam imposes no additional burden on cardiovascular system when a infusion of large dose of lidocaine is given to dogs anesthetized with halothanenitrous oxide. But caution may be advised if the addition of lidocaine is indicated in subjects who have impared autonomic nervous system and who are in hypercarbic, hypoxic, or acidotic states.

• Journal of Chest Surgery
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• v.38 no.5 s.250
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• pp.349-356
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• 2005

Background: Arterial conduits using in coronary artery bypass grafting (CABG) have been known a great long term patency rates, and improved short and long term clinical outcomes. It has been reported that Off pump CABG has better clinical results than CABG using cardiopulmonary bypass. To evaluate the advantage of arterial conduits over venous conduits and to avoid the adverse effects of cardiopulmonary bypass, we performed total arterial Off pump CABG. Material and Method: From January 2001 to October 2004, Off pump CABG using only arterial conduits was performed on 325 patients with a mean age of $59.3\pm11.9$ years ($36\~83$). Mean ejection fraction was $55.4\pm14.0\%\;(15\~86).$ Angiography showed left main disease or triple-vessel disease in $81.9\%$ of the patients. Indications of using arterial conduits was stenosis $\ge50\%$ of left anterior descending artery, stenosis $\ge80\%$ of branches of left circumflex artery, and stenosis $\ge90\%$ of right coronary artery and its branches. Multi-slice computed tomography was performed on 194 patients to evaluate the short term patency rates. Result: A total of 928 distal anastomoses were performed and the average anastomoses per a patient were $2.86\pm0.78$. There was 1 operative mortality. Postoperative complications were mediastinitis in 6 patients ($1.8\%$), renal failure in 4 patients ($1.2\%$), perioperative myocardial infarction in 3 patients ($0.9\%$), reoperation for bleeding in 3 patients ($0.9\%$). There was no postoperative stroke. Patency rate of arterial conduits was $99.3\%$ (581/585). There were 4 stenoses or competitive flows in 2 radial arteries and 2 right internal mammary arteries. Conclusion: Total arterial Off pump CABG appears to be safe, showing a low surgical mortality and morbidity and excellent short term patency rates of arterial conduits.

• Journal of Oral Medicine and Pain
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• v.31 no.1
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• pp.1-16
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• 2006

The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.