Although various automated CBC analyzers with different WBC analytical principles were consequently introduced to clinical laboratory, the specific information concerning the suitability or unsuitability of aging samples is scarce. For this reason, we studied the effect of storage duration and temperature on CBC parameter in SE-9000 (SYSMEX Medical Electronics Co., Ltd., Kobe, Japan), automated CBC analyzer. We tested 32 K3-EDTA specimens with SE-9000 during 72 hours. Specimens were kept at room temperature (RT) and refrigerated and were analyzed at 0 hr, 4 hr, 8 hr, 24 hr, 48 hr and 72 hr after the collection of the specimens. The percentage changes from initial value for each parameters were calculated. Among the CBC parameters, hemoglobin, red blood cell count, mean corpuscular hemoglobin and platelets were stable for the study period at both temperatures. The mean corpuscular volume (MCV), hematocrit (Hct) and red cell distribution (RDW) increased and the mean corpuscular hemoglobin concentration (MCHC) decreased over time at room temperature. These parameters were stable when refrigerated. The leukocyte count was stable during 72hr at RT and when refrigerated. At room temperature, the relative percentages of neutrophils tend to increase, whereas those of lymphocyte and monocytes tend to decrease after 48 hours. When refrigerated, those of neutrophils and monocytes tend to increase, whereas those of lymphocytes tend to decreased over time. CBC parameters of refrigerated specimen were reliable for 72 hr for the exception of differential count from 24 hr but many CBC parameters, such as MCV, Hct, MCHC, RDW and differential count of leukocyte of blood stored at room temperature for 24 hr were unreliable.

The purpose of this study was to evaluate the usefulness of the automated TB-PCR assay for the detection of Mycobacterium tuberculosis. The 807 cases were analyzed with their TB-PCR, AFB smear and culture in bronchial washing fluids, sputum and body fluids samples. The TB-PCR positive of the bronchial washing fluid, sputum and body fluids were 11.3%, 7.3% and 3.6%, respectively, in cases of AFB smear-negative and culture-negative. The sensitivity values of the bronchial washing fluid, sputum and body fluids were 93.3%, 100% and 50%, respectively, according to the culture result. The sensitivity of body fluids was lower than that of the bronchial washing fluid and sputum. The specificity values of the bronchial washing fluid, sputum and body fluids were 83.3%, 89.0% and 95.7%, respectively, according to the culture result. In conclusion, the automated TB-PCR assay proved to be a useful method for the detection of Mycobacterium tuberculosis in the bronchial washing fluid and sputum. But we think that there is still a need for us to study body fluids further.

Although Mycobacterium tuberculosis complex strains remain responsible for the majority of diseases caused by mycobacterial infections worldwide, the increase in HIV infections has allowed for the emergence of other non-tuberculous mycobacteria as clinically significant pathogens. However, Mycobacterium species has a long period of incubation, and requires serious biochemical tests such as niacin, catalase, and nitrate test that are often tedious. The development of rapid and accurate diagnostics can aid in the early diagnosis of disease caused by Mycobacterium. The current DNA amplification and hybridization methods that have been developed target several genes for the detection of mycobacterial species such as hps65, 16S rDNA, rpoB, and dnaj. These methods produce rapid and accurate results. In this study, PCR-restriction fragment length polymorphism analysis(PCR-RFLP) based on the region of the rpoB gene was used to verify the identification of non-tuburculosis Mycobacterium species. A total of 8 mycobacterial reference strains and 13 clinical isolates were digested with restriction enzymes such as Msp I in this study. The results of using this process clearly demonstrated that all 13 specimens were identified by rpoB gene PRA method. The PCR-RFLP method based on the rpoB gene is a simple, rapid, and accurate test for the identification of Mycobacterium.

Metallo-${\beta}$-lactamase (MBL) can hydrolyze all ${\beta}$-lactams except monobactams and frequently coexists with various antibiotic resistance genes such as aminoglycoside resistance, sulfonamide resistance gene, etc. Therefore, the effective antibiotics against infections by these bacteria are markedly limited or can't even be found. We tried to search in-vitro antimicrobial combinations with synergistic effects for a VIM-2 type MBL producing Pseudomonas aeruginosa, isolated from clinical specimen. On the selection of antibiotic combinations with synergistic effects, we performed a one disk synergy test, modified Pestel's method, in agar without aztreonam (AZT). The bacteriostatic synergistic effects of this tests were scored as $S_1$ (by susceptibility pattern in agar without antibiotics), $S_2$ (by the change of susceptibility in agar with or without antibiotics) and $S_3$ ($S_1$ + $S_2$) and was classified into weak (1 point), moderate (2 points) and strong (3 points) by $S_3$ score. Subsequently, we carried out the time-killing curve for the antibiotic combinations with the strong synergistic bacteriostatic effect. One VIM-2 type MBL producing P. aeruginosa confirmed by the PCR showed all resistance against all ${\beta}$-lactams except AZT, aminoglycoside and ciprofloxacin. In the one disk synergy test, this isolate showed a strong bacteriostatic synergistic effect for the antibiotic combination of AZT and piperacillin-tazobactam (PIP-TZP) or AZT and amikacin (AN). On the time-killing curve after six hours of incubation, the colony forming units (CFUs/mL) of this bacteria in the medium broth with both combination antibiotics were decreased to 1/18.7, 1/17.1 of the least CFUs of each single antibiotics. The triple antibiotic combination therapy including AZT, PIP-TZP and AN was shown to be significantly synergistic after 8 hrs of exposure. In a VIM-2 MBL producing P. aeruginosa with susceptibility for AZT, the triple antibiotic combination therapy including AZT, PIP-TZP and AN may be considered as an alternative antibiotics modality against the infection by some MBL type. But the antimicrobial combination therapy for many more MBL producing isolates is essential to know as soon as possible for the selection of effective treatment against the infection by this bacteria.

The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) is the main mechanism of bacterial resistance to third-generation cephalosporins and monobactams, whose prevalence varies depending on the different geographical areas. In the last years it has increased notably to the point of being considered a health problem of great importance. The characterization of the ESBLs producing Klebsiella penumoniae strains present in clinical isolates is time-consuming. I describe here the development of a new system, which consists of a multiplex PCR. I found 51 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disc synergy test showed 47 positive K. pneumoniae, which were K. pneumoniae isolates. All ESBLs producing K. pneumoniae strains were resistant to antibiotic amikacin, gentamicin and ciprofloxacin. By multiplex PCR analysis, $bla_{TEM}$ gene in 17 strains 44 $bla_{SHV}$ genes and $bla_{CTX}$ genes in 33 strains were identified. In this study, the multiplex polymerase chain reaction (PCR) assay was a good method to detect and differentiate ESBLs producing K. penumoniae strains in clinical isolates.

Alzheimer's disease (AD) is characterised neuropathologically by the accumulation of neuritic plaques and neurofibrillary tangles as well as by cerebrovascular amyloid deposition and neuronal cell loss. Current advances have shown the apolipoproteinE-epsilon 4 (ApoE4) allele to be highly associated with late-onset familial and sporadic Alzheimer's disease (AD) in Western populations. The association of ApoE allele frequencies and dementia remains unknown in populations from many countries. We recently initiated a project to examine ApoE frequencies in non-demented healthy Koreans. Genomic DNA in hair root from a thousand persons was collected and ApoE gene type was investigated with the methods of polymerase chain reaction (PCR) and restriction fragment length polymorphism. A group of a thousand non-demented Koreans over the age of 40 years were found to be positive in 15.7% of the cases for ApoE4. AD and ApoE4 were closely related. ApoE epsilon 4 was a dangerous factor of AD and ApoE 4 allele made a contribution to the heterogenicity of AD.

The purpose of the recovery experiment in clinical chemistry is performed to estimate proportional systematic error. We must know all measurements have some error margin in measuring analytical performance. Proportional systematic error is the type of error whose magnitude increases as the concentration of analyte increases. This error is often caused by a substance in the sample matrix that reacts with the sought for analyte and therefore competes with the analytical reagent. Recovery experiments, therefore, are used rather selectively and do not have a high priority when another analytical method is available for comparison purposes. They may still be useful to help understand the nature of any bias revealed in the comparison of kit experiments. Recovery should be expressed as a percentage because the experimental objective is to estimate proportional systematic error, which is a percentage type of error. Good recovery is 100.0%. The difference between 100 and the observed recovery(in percent) is the proportional systematic error. We calculated the amount of analyte added by multiplying the concentration of the analyte added solution by the dilution factor(mL standard)/(mL standard + mL specimen) and took the difference between the sample with addition and the sample with dilution. When making judgments on method performance, the observed that the errors should be compared to the defined allowable error. The average recovery needs to be converted to proportional error(100%/Recovery) and then compared to an analytical quality requirement expressed in percent. The results of recovery experiments were total protein(101.4%), albumin(97.4%), total bilirubin(104%), alkaline phosphatase(89.1%), aspartate aminotransferase(102.8), alanine aminotransferase(103.2), gamma glutamyl transpeptidase(97.6%), creatine kinase(105.4%), lactate dehydrogenase(95.9%), creatinine(103.1%), blood urea nitrogen(102.9%), uric acid(106.4%), total cholesterol(108.5), triglycerides(89.6%), glucose(93%), amylase(109.8), calcium(102.8), inorganic phosphorus(106.3%). We then compared the observed error to the amount of error allowable for the test. There were no items beyond the CLIA criterion for acceptable performance.

Behcet's disease (BD) is a chronic, multisystemic disorder which is more frequently seen in the Mediterranean basin, Middle East, and Far East. The causes and pathogenesis of BD are unknown although many possibilities are being investigated. The diagnosis of BD is based on clinical manifestations because there are no pathognomonic laboratory tests. So, the purpose of this study was to examine the prevalence of HLA-B51 in patients with BD. We used the whole blood of 33 patients diagnosed with BD at Chosun University Hospital from August 2003 to January 2006. For the HLA-B51 test, we extracted the DNA from the whole blood of 33 BD patients, and we investigated it through the nested PCR method. Data were analyzed using the SPSS/PC 10.0. The frequencies of gender of the 33 cases diagnosed as BD were male 13 (39.4%) and female 20 (60.6%). The frequencies of age group of the 33 cases diagnosed as BD were 20 yrs 8 (24.2%), 30 yrs 12 (36.4%), 40 yrs 8 (24.2%), 50 yrs 1 (3.0%), and 60 yrs and 70 yrs 2 (6.1%), respectively. The frequencies of HLA-B51 of the 33 cases diagnosed as BD were HLA-B51-negative 18 (54.5%) and HLA-B51-positive 15 (45.5%). In conclusion, BD occurred more often in women than men (1: 1.53), and the mean age of the BD patients was 39.8 years old. HLA-B51 was positive in 45.5% of patients with BD, and was statistically significant in age (p<0.05).

최근 만성 B형간염의 치료에 B형간염 바이러스 복제를 저해하여 감염력을 약화시키는 lamivudine이 많이 사용되고 있다. 그러나 이 약제를 장기간 복용할 경우 B형간염 바이러스 DNA polymerase 유전자의 YMDD motif에 아미노산 치환을 일으켜 lamivudine 저항성 B형간염 바이러스가 나타나는 점이 문제시 되고 있다. 본 연구의 목적은 lamivudine 치료를 받은 만성 B형간염 환자에서 PCR-direct sequencing 법을 이용하여 B형간염 바이러스 DNA 중합효소 유전자의 YMDD motif(codon 552)와 codon 528에서 돌연변이 출현빈도를 구하고, HBeAg과의 관련성을 보고자 하였다. 방법은 만성 B형간염 환자의 혈청에서 DNA를 추출하고 DNA 중합효소 유전자의 codon 552와 528을 포함하는 부위에서 선택한 두 쌍의 primer를 이용하여 nested PCR을 실시하였다. 증폭된 PCR 산물은 2% agarose gel에서 전기영동을 한 후, 자동염기서열분석기를 이용하여 sequencing을 실시하였다. 총 207명 중에서 돌연변이는 124명(60%)에서 발견되었으며, 남, 녀에서 차이는 발견되지 않았고, 돌연변이군에서 비돌연변이군에 비해 평균나이가 약간 높게 나타났으나 유의성은 없었다. Codon 552에서 단일돌연변이로는 M552I(50.8%)가 가장 높게 나타났고, 다음으로 M552V(43.5%), M552I/V(5.7%)의 순서로 나타났다. Codon 528에서는 67.0%의 L528M 돌연변이가 발견되었다. Codon 552와 codon 528에서 동시에 발생한 중복돌연변이로는 M552V/L528M(43.6%)이 가장 높게 나타났고, 다음으로 M552I/L528(33.1%) 그리고 M552I/L528M(17.7%)의 순으로 나타났다. Codon 552에서 serine 돌연변이(M528S)는 발견되지 않았으며, L528M은 M552V 돌연변이와 거의 동시에 검출되었다. 본 연구에서 만성 B형간염환자에서 HBeAg의 유무와 lamivudine 돌연변이율과의 상관성은 발견되지 않았으며, PCR-direct sequencing법은 고가의 자동염기서열분석 장비와 숙련된 기술자가 필요하다는 문제점은 있으나, 검체 수가 많은 큰 임상검사실에서는 활용성이 클 것으로 판단된다. 향 후 lamivudine으로 인한 HBV 돌연변이형과 환자의 임상결과의 관련성에 대한 연구가 추가적으로 실시되면, lamivudine을 복용하는 만성 HBV 환자의 치료와 예후에 유용할 것으로 사료된다.

The purpose of this study was to develop discriminant analysis models for predicting cervical normal/dysplasia case diagnoses using cytometric features derived from the digital image analysis of cell monolayers. The database consisted of 19 cases diagnosed either as normal (n=5), moderate dysplasia (n=7), severe dysplasia (n=7) on monolayer preparations. We studied the nuclear and cytoplasmic characteristics of cells in the normal, moderate dysplasia and severe dysplasia on cervical samples. The morphometric parameters selected for the analysis were nuclear/cytoplasmic ratio and the nuclear variations measured by image analysis on normal and precancerous lesions of cervical smears; several shape factors; area; perimeter; maximal, minimal and equivalent circle diameters. The results showed that the dysplasia samples exhibited changes in both cellular and nuclear form and size but lacked substantial differences in the tumor grades. The coefficient of nuclear variation is as follows to normal cell $21.8{\pm}3.2%$, moderate dysplasia $33.5{\pm}6.1%$, severe dysplasia $27.7{\pm}5.8$ of cervical smears.

Microalbuminuria is most frequently caused by kidney damage from diabetes. Moreover, many other conditions can lead to kidney damage, such as high blood pressure, heart failure, cirrhosis, or systemic lupus erythematosus (SLE). The measurement of the microalbumin in urine may be useful for the early diagnosis or as a predictor of nephropathy in diabetes. The most common method for getting a quantitative measurement of urinary protein relies on a 24-hour urine collection. The result of this method is accurate. But 24hr urine collection is difficult to obtain and variations in volume are frequent. Also the patients complain about urine collection. We tried to measure reference values for microalbumin using fasting urine and compare them with the albumin/creatinine ratio using 24hr urine. The concentrations of microalbumin in fasting urine and 24hr urine were $7.1{\pm}3.8mg/L$, $5.7{\pm}2.9mg/L$ (r=0.61, p=0.27), respectively. The albumin/creatinine ratios using fasting urine and 24hr urine were $8.7{\pm}4.2{\mu}g/mg$, $8.7{\pm}4.0{\mu}g/mg$ (r=0.76, p=0.88), respectively. This study indicated that the measurement of microalbumin in fasting urine was an easy and simple method for early diagnosis or to predict nephropathy in diabetes. Thus, setting up the reference value using fasting urine may be useful in the screening test for the diabetic nephropathy patients instead of using the 24hr albumin excretion rate (AER).

Obstructive sleep apena syndrome (OSAS) is accompanied by the following symptoms: apnea caused by upper respiratory tract obstruction while sleeping, repetitive lowering of $SpO_2$, severely affected excessive daytime sleepiness (EDS), suffocation/frequent awakeness while sleeping, daytime lethargy, and lack of concentration. OSAS was investigated with sex, age, body weight, body mass index (BMI), neck circumference and snoring sound as clinical characteristics and the anticipating factors of OSAS were studied in relation with the apnea-hypopnea index (AHI). The subjects were 42 people (male 34/female 8) who visited the clinic due to snoring and had polysomnography evaluation. AHI was differenciated into normal (less than 0~5/hr), mild (5~15/hr), moderate (15~30/hr) and severe (more than 30/hr). As the apnea-hypopnea index (AHI) gets higher, the snoring sound was louder (p<0.01), neck circumference was thicker (p<0.05) and also there were relative correlations with body weight (p<0.01), body mass index (p<0.05), snoring sound (p<0.01) and neck circumference (p<0.01). Since the snoring sound and neck circumference explained 32.8% of the AHI distribution, if the patient was severely snoring or had a thick neck circumference due to obesity, the apnea-hypopnea index showed a predisposition to the obstructive sleep apnea syndrome.

Sensorineural hearing loss is frequently found in patients with chronic renal failure (CRF). There have been many efforts to elucidate the etiologic factors of hearing loss in patients with CRF. However, there was not any clear identified cause of hearing loss. This study was undertaken to evaluate the activity of mitochondrial respiratory chain (MRC) in CRF patients with hearing impairment. To determine MRC activity, peripheral blood cells were obtained from CRF patients with hearing impairment receiving dialysis and normal subjects without any hearing problems. MRC activity of complex I and complex III was measured by the Trounces method. In MRC activities between the normal subjects group and CRF patients with hearing problems, the complex I and III activities of CRF patients with hearing problems were 63% and 85% compared with normal subjects (p<0.01). These results suggest that the activity of MRC may be implicated in the underlying mechanism of the hearing impairment in CRF patients, through mitochondrial DNA mutations at MRC complex I region with a decrement of MRC activity.

Up to now, there have been rare clinical studies on leaders and aerobics athletes. To get the useful data for protecting from auditory disorder, we selected 15 female aerobics leaders (experimental group) and 15 females (control group) unexperienced in aerobics and a without neurological and octolaryngological disorder. The average age was $34.87{\pm}8.80$ (experimental group) and $34.07{\pm}8.45$ (control group) years, and the average career of an aerobics leader (experimental group) was $8.33{\pm}4.73$ years. We measured the auditory evoked potential (AEP) of the two groups treated with 70, 75 and 85 dB intensity from January 2006 to May 2006 and analyzed the absolute latency (AL) and interpeak latency (IPL) by the SPSS/pc+ 12.0 program. In the comparison of the AL between the experimental group and the control group according to intensity, both ears treated with 70 and 75 dB had a significant difference (p<0.05) in the I, III, V wave and in the I, V wave respectively, and the experimental group treated with 85 dB showed a difference in the I, III, V wave (left ear) and in the I wave (right ear) respectively. The IPL of the two groups treated with various intensities had no prolongation. In the comparison of the AL between the experimental group and the control group according to ages, the experimental group in their 20s treated with 70 dB showed a significant difference (p<0.05) in the V wave (left ear) and in the I, III, V wave (right ear), and the experimental group in their 20s treated with 75 dB in the I, III wave (left ear) and in I, III, V wave (right ear), and experimental group in their 20s treated with 85 dB in the V wave (left ear) and in the III, V wave (right ear). The experimental group in their 30s treated with 70 dB had a significant difference (p<0.05) only in the V wave (right ear). Only in the IPL of subjects in their 20s treated with 85 dB, III-V and I-V of both ears was extended. In the comparison of the AL and IPL according to career, there was no significant difference between the two groups. From this results, we concluded that the lower sound intensity (70 dB) showed a more significant difference in the experimental group than the control group. We concluded that the leader of aerobics exposed to louder sounds than normal people are affected by auditory neurological and octolaryngological disorders. So we think that the leaders of aerobics need to control the noise level for protecting themselves against a disease.