Clinical Implications of Quantitative JAK2 V617F Analysis using Droplet Digital PCR in Myeloproliferative Neoplasms

BACKGROUND: JAK2 V617F is the most common mutation in myeloproliferative neoplasms (MPNs) and is a major diagnostic criterion. Mutation quantification is useful for classifying patients with MPN into subgroups and for prognostic prediction. Droplet digital PCR (ddPCR) can provide accurate and reproducible quantitative analysis of DNA. This study was designed to verify the correlation of ddPCR with pyrosequencing results in the diagnosis of MPN and to investigate clinical implications of the mutational burden. METHODS: Peripheral blood or bone marrow samples were obtained from 56 patients newly diagnosed with MPN or previously diagnosed with MPN but not yet indicated for JAK2 inhibitor treatment between 2012 and 2016. The JAK2 V617F mutation was detected by pyrosequencing as a diagnostic work-up. The same samples were used for ddPCR to determine the correlation between assays and establish a detection sensitivity cut-off. Clinical and hematologic aspects were reviewed. RESULTS: Forty-two (75%) and 46 (82.1%) patients were positive for JAK2 V617F by pyrosequencing and ddPCR, respectively. The mean mutated allele frequency at diagnosis was 37.5±30.1% and was 40.7±31.2% with ddPCR, representing a strong correlation (r=0.9712, P < 0.001). Follow-up samples were available for 12 patients, including eight that were JAK2 V617F-positive. Of these, mutational burden reduction after treatment was observed in six patients (75%), consistent with trends of hematologic improvement. CONCLUSIONS: Quantitative analysis of the JAK2 V617F mutation using ddPCR was highly correlated with pyrosequencing data and may reflect the clinical response to treatment.

Using Array-Based Comparative Genomic Hybridization to Diagnose Pallister-Killian Syndrome

Pallister-Killian syndrome (PKS) is a rare multisystem disorder characterized by isochromosome 12p and tissue-limited mosaic tetrasomy 12p. In this study, we diagnosed three pediatric patients who were suspicious of having PKS using array-based comparative genomic hybridization (array CGH) and FISH analyses performed on peripheral lymphocytes. Patients 1 and 2 presented with craniofacial dysmorphic features, hypotonia, and a developmental delay. Array CGH revealed two to three copies of 12p in patient 1 and three copies in patient 2. FISH analysis showed trisomy or tetrasomy 12p. Patient 3, who had clinical features comparable to those of patients 1 and 2, was diagnosed by using FISH analysis alone. Here, we report three patients with mosaic tetrasomy 12p. There have been only reported cases diagnosed by chromosome analysis and FISH analysis on skin fibroblast or amniotic fluid. To our knowledge, patient 1 was the first case diagnosed by using array CGH performed on peripheral lymphocytes in Korea.

Thyroxine (T4) Autoantibody Interference of Free T4 Concentration Measurement in a Patient With Hashimoto's Thyroiditis

No abstract available.

Association between Body Mass Index, Waist Circumference and Prevalence of Microalbuminuria in Korean Adults of Age 30 Years and Older without Diabetes, Hypertension, Renal Failure, or Overt Proteinuria: The 2013 Korean National Health and Nutrition Exami / 가정의학회지

BACKGROUND: Microalbuminuria and obesity markers are known risk factors for cardiovascular or renal disease. This study aimed to evaluate the prevalence of microalbuminuria according to body mass index (BMI) and abdominal obesity criteria. METHODS: The study subjects included 3,979 individuals aged 30 years or older who did not have diabetes, hypertension, renal failure, or overt proteinuria, from among those who participated in The Korean National Health and Nutrition Examination Survey in 2013, a cross-sectional, nationally representative, stratified survey. Microalbuminuria was defined as a urinary albumin to creatinine ratio of 30 to 300 mg/g. BMI and waist circumference were classified according to the Asia-Pacific criteria. RESULTS: The prevalence of microalbuminuria was found to be 5.1%. In the normoalbuminuria group, 3.4%, 41.7%, 24%, 27.6%, and 3.2% of participants were included in the underweight, normal, overweight, obesity 1, and obesity 2 groups, respectively. These percentages in the microalbuminuria group were 7.1%, 34.5%, 19.2%, 28.6%, and 10.6%, respectively (P<0.001). The waist circumference in men was 21.4% in the normoalbuminuria group and 36.5% in the microalbuminuria group (P=0.004). Logistic regression analyses were performed to evaluate the relationship between the presence of microalbuminuria and BMI or waist circumference groups. The risk of microalbuminuria was significant only in the underweight group (odds ratio, 13.22; 95% confidence interval, 2.55-68.63; P=0.002) after adjusting for confounding factors, abdominal obesity was not significantly associated with microalbuminuria. CONCLUSION: The prevalence of microalbuminuria in a general population in Korea was associated with underweight in men and was not associated with waist circumference in either men or women.

Development and Validation of Liquid Chromatography-Tandem Mass Spectrometry Method for Quantification of Plasma Metanephrines for Differential Diagnosis of Adrenal Incidentaloma

High-resolution imaging techniques have increased the detection rate of adrenal incidentaloma. We developed a method of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for detection of plasma free metanephrine (MN) and normetanephrine (NMN) and evaluated its analytical performance and clinical efficacy in differential diagnosis of adrenal incidentaloma. After solid-phase extraction, chromatographic isolation of the analytes and internal standard was achieved by column elution in the LC-MS/MS system. The analytes were detected in multiple-reaction monitoring mode by using positive electrospray ionization: MN, transition m/z 180.1-->165.1; NMN, m/z 166.1-->134.1. This method was validated for linearity, precision, accuracy, lower limits of quantification and detection, extraction recovery, and the matrix effect. Plasma concentrations of MN and NMN of 14 patients with pheochromocytoma were compared with those of 17 healthy volunteers, 10 patients with essential hypertension, and 60 patients with adrenal adenoma. The assay's linear range was 0.04-50.0 and 0.08-100.0 nmol/L for MN and NMN, respectively. Assay imprecision was 1.86-7.50%. The accuracy ranged from -7.50% to 2.00%, and the mean recovery of MN and NMN was within the range 71.5-95.2%. Our LC-MS/MS method is rapid, accurate, and reliable and useful for differential diagnosis of adrenal incidentaloma.

Dried Blood Spot Testing for Seven Steroids Using Liquid Chromatography-Tandem Mass Spectrometry With Reference Interval Determination in the Korean Population

BACKGROUND: Conventional screening for congenital adrenal hyperplasia (CAH) using immunoassays generates a large number of false-positive results. A more specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been introduced to minimize unnecessary follow-ups. However, because of limited data on its use in the Korean population, LC-MS/MS has not yet been incorporated into newborn screening programs in this region. The present study aims to develop and validate an LC-MS/MS method for the simultaneous determination of seven steroids in dried blood spots (DBS) for CAH screening, and to define age-specific reference intervals in the Korean population. METHODS: We developed and validated an LC-MS/MS method to determine the reference intervals of cortisol, 17-hydroxyprogesterone, 11-deoxycortisol, 21-deoxycortisol, androstenedione, corticosterone, and 11-deoxycorticosterone simultaneously in 453 DBS samples. The samples were from Korean subjects stratified by age group (78 full-term neonates, 76 premature neonates, 89 children, and 100 adults). RESULTS: The accuracy, precision, matrix effects, and extraction recovery were satisfactory for all the steroids at three concentrations; values of intra- and inter-day precision coefficients of variance, bias, and recovery were 0.7-7.7%, -1.5-9.8%, and 49.3-97.5%, respectively. The linearity range was 1-100 ng/mL for cortisol and 0.5-50 ng/mL for other steroids (R2>0.99). The reference intervals were in agreement with the previous reports. CONCLUSIONS: This LC-MS/MS method and the reference intervals validated in the Korean population can be successfully applied to analyze seven steroids in DBS for the diagnosis of CAH.

Evaluation of the Analytical Performance of a Direct Quantitative Assay of Small Dense LDL / 임상검사와정도관리

BACKGROUND: Small dense low density lipoprotein (sdLDL) plays a critical role in the progression of coronary vascular disease. However, regardless of the accuracy of the analytical technique, routine measurement of LDL does not precisely ascertain LDL particle size. Therefore, we evaluated the performance of a direct quantitative assay of sdLDL that combines a precipitation method with filtration (Denka Seiken, Japan). METHODS: We evaluated the precision, linearity, carry-over, and sample stability of a sdLDL reagent. A reference interval was established, and method comparison was performed with the Lipoprint LDL system using polyacrylamide gel tube electrophoresis (Quantimetrix, USA). RESULTS: The within-run precision was 0.9% to 1.4%, with a total precision of 3.2% to 3.5%. The analytical measurement ranged from 4.1 to 101.3 mg/dL. The calculated carry-over was negligible (0.1%). Based on a comparison conducted using the Lipoprint LDL system, the median sdLDL concentration of 57 individuals with phenotype non-A was found to be significantly higher than that of 51 subjects with phenotype A (43 vs. 22 mg/dL, P<0.0001). The levels in samples retested after storage showed more than 95% recovery when stored in a refrigerator (5degrees C) for 2 weeks and at -20degrees C or lower for 4 weeks. The reference interval of sdLDL was between 7.6 and 52.0 mg/dL. CONCLUSIONS: This method of sdLDL measurement showed good performance and can be easily applied to automated analysers in clinical laboratories.

Identification of a De Novo Heterozygous Missense FLNB Mutation in Lethal Atelosteogenesis Type I by Exome Sequencing

BACKGROUND: Atelosteogenesis type I (AO-I) is a rare lethal skeletal dysplastic disorder characterized by severe short-limbed dwarfism and dislocated hips, knees, and elbows. AO-I is caused by mutations in the filamin B (FLNB) gene; however, several other genes can cause AO-like lethal skeletal dysplasias. METHODS: In order to screen all possible genes associated with AO-like lethal skeletal dysplasias simultaneously, we performed whole-exome sequencing in a female newborn having clinical features of AO-I. RESULTS: Exome sequencing identified a novel missense variant (c.517G>A; p.Ala173Thr) in exon 2 of the FLNB gene in the patient. Sanger sequencing validated this variant, and genetic analysis of the patient's parents suggested a de novo occurrence of the variant. CONCLUSIONS: This study shows that exome sequencing can be a useful tool for the identification of causative mutations in lethal skeletal dysplasia patients.

A Case of Catheter-related Kocuria marina Bloodstream Infection in a Patient with Multiple Myeloma

Kocuria species are gram-positive cocci that are commonly found on human skin, mucosa, and oropharynx. They can cause opportunistic infections in patients with underlying disease or intravenous catheterization. Although several cases of bacteremia caused by Kocuria rosea, Kocuria kristinae, and Kocuria rhizophila have been documented, K. marina has not been reported as a bloodstream pathogen. We isolated K. marina from the blood of a patient with multiple myeloma as a cause of catheter-related bacteremia. The VITEK II system (bioMerieux, Inc.) identified the isolate as K. variance. However, 16S rRNA gene sequencing analysis identified the isolate as K. marina. Accurate identification of Kocuria to the species level requires molecular methods that precisely identify these rare pathogens.

Successful Azathioprine Treatment with Metabolite Monitoring in a Pediatric Inflammatory Bowel Disease Patient Homozygous for TPMT*3C

Thiopurine S-methyltransferase (TPMT) methylates purine analogues, showing TPMT activity in inverse relation to concentrations of active metabolites such as 6-thioguanine nucleotide (6-TGN). With conventional dosing of thiopurines, patients with homozygous variant TPMT alleles consistently suffer from severe myelosuppression. Here, we report a patient with TPMT*3C/*3C who managed successfully with monitoring of thiopurine metabolites. The patient was an 18-year-old male diagnosed with Crohn's disease. The standard dose of azathioprine (AZA) (1.8 mg/kg/day) with mesalazine (55.6 mg/kg/day) was prescribed. Two weeks after starting AZA treatment, the patient developed leukopenia. The DNA sequence analysis of TPMT identified a homozygous missense variation (NM_000367.2: c.719A>G; p.Tyr240Cys), TPMT*3C/*3C. He was treated with adjusted doses of azathioprine (0.1-0.2 mg/kg/day) and his metabolites were closely monitored. Leukopenia did not reoccur during the follow-up period of 24 months. To our knowledge, this is the first case of a patient homozygous for TPMT*3C successfully treated with azathioprine in Korea. While a TPMT genotyping test may be helpful to determine a safe starting dose, it may not completely prevent myelosuppression. Monitoring metabolites as well as routine laboratory tests can contribute to assessing drug metabolism and optimizing drug dosing with minimized drug-induced toxicity.

A Novel UMOD Mutation (c.187T>C) in a Korean Family with Juvenile Hyperuricemic Nephropathy

Familial juvenile hyperuricemic nephropathy (FJHN; OMIM 162000) is an autosomal dominant disorder characterized by hyperuricemia and gouty arthritis due to reduced kidney excretion of uric acid and progressive renal failure. Gradual progressive interstitial renal disease, with basement membrane thickening and glomerulosclerosis resulting from fibrosis, starts in early life. In most cases of FJHN, uromodulin gene (UMOD) is responsible for the disease; however, there has been only one report of a genetically confirmed FJHN family in Korea. Here we report another Korean family with FJHN, in which three male members. a father and 2 sons.developed gout and progressive renal insufficiency. The clinical, laboratory, and radiological findings were consistent with FJHN, and renal biopsy showed chronic parenchymal damage, which can be found in FJHN but is not specific to this disease. In order to confirm the diagnosis, sequence analysis of the UMOD was performed, and a novel heterozygous missense variant (c.187T>C; p.Cys63Arg) in exon 3 was identified. We assume that this variant is likely to be the causative mutation in this family, as the variant segregated with the disease. In addition, approximately two-thirds of the known mutations lead to a cysteine amino acid change in uromodulin, and all such variants have been shown to cause UMOD-associated kidney disease. In summary, we report a Korean FJHN family with three affected members by genetic analysis of the UMOD, and provide the first report of a novel heterozygous missense mutation.

Mycoplasma pneumoniae infection in patients with Kawasaki disease / 소아과

PURPOSE: Kawasaki disease (KD) is the main cause of acquired heart disease in children. In addition to cardiovascular involvement, many complications have been recognized in KD. However, respiratory complications have been rarely reported. We investigated the differences in clinical characteristics, laboratory findings, radiography findings, and echocardiography findings of Mycoplasma pneumoniae infection and other types of pneumonia in KD patients. METHODS: Among 358 patients with KD, 54 developed concurrent pneumonia. Among the 54 patients, 12 (22.2%) with high titers of anti-M. pneumoniae antibody (AMA) (>1:640) were grouped in the M. pneumoniae group and 42 were included in the control group. Serum AMA was measured in each patient. Clinical laboratory findings and total duration of fever were analyzed. RESULTS: The duration of fever, serum hemoglobin, white blood cell count, platelet count, erythrocyte sedimentation rate, C-reactive protein level, albumin level, and the incidence of coronary arterial lesions showed no statistical difference in the 2 groups. Neutrophil count was significantly higher in the M. pneumoniae group than in the control group. Among various radiography findings observed in pneumonia, consolidation and pleural effusion were more frequent in the M. pneumoniae group than in the control group. On the other hand, parahilar peribronchial opacification, diffuse interstitial lesion, and normal findings prevailed in the control group. CONCLUSION: KD patients can have concurrent infections, especially pulmonary symptoms. The cause of KD is likely to be associated with M. pneumoniae infection. Thus, immediate treatment of M. pneumoniae infection in KD patients is very important.

Usefulness of the Waist Circumference-to-Height Ratio in Screening for Obesity in Korean Children and Adolescents / 대한소아소화기영양학회지

PURPOSE: To evaluate the feasibility and usefulness of the waist circumference-to-height ratio (WHTR) in screening for obesity in Korean children and adolescents. METHODS: Data, including body mass index (BMI), waist circumference (WC), and height, were obtained from the national growth surveys for children and adolescents in 2005. The WHTR was calculated dividing WC by height in subjects 2~18 years of age. Overweight and obese were defined by BMI percentiles for age and gender. The receiver operating characteristic (ROC) analysis was performed to find out the optimal cutoff values of WHTR that matched BMI-determined overweight and obesity using the STATA program. The area under the curve (AUC), a measure of diagnostic power, of WHTR was compared to WC. The influence of age on WHTR was analyzed by the SAS program. RESULTS: The WHTR significantly decreased with age, and had less correlation with age in the 6~18-year-old age group than the 2~5-year-old age group. Furthermore, the WHTR also had less correlation with age than WC in the 6~18-year-old age group. The AUC of WHTR in identifying overweight and obesity was significantly higher than the AUC of WC in the 6~18-year-old age group. The optimal cutoff values were 0.51 in boys and 0.49 in girls for obesity, and 0.48 in boys and 0.47 in girls for overweight, with all having the AUC>0.9. The optimal cutoff values of WHTR had a higher sensitivity for diagnosing obesity than WC> or =90th percentiles. CONCLUSION: The WHTR is an easy, accurate, and less age-dependent index with high applicability in screening for obesity in children and adolescents.

Usefulness of the Waist Circumference-to-Height Ratio in Screening for Obesity in Korean Children and Adolescents / 대한소아소화기영양학회지

PURPOSE: To evaluate the feasibility and usefulness of the waist circumference-to-height ratio (WHTR) in screening for obesity in Korean children and adolescents. METHODS: Data, including body mass index (BMI), waist circumference (WC), and height, were obtained from the national growth surveys for children and adolescents in 2005. The WHTR was calculated dividing WC by height in subjects 2~18 years of age. Overweight and obese were defined by BMI percentiles for age and gender. The receiver operating characteristic (ROC) analysis was performed to find out the optimal cutoff values of WHTR that matched BMI-determined overweight and obesity using the STATA program. The area under the curve (AUC), a measure of diagnostic power, of WHTR was compared to WC. The influence of age on WHTR was analyzed by the SAS program. RESULTS: The WHTR significantly decreased with age, and had less correlation with age in the 6~18-year-old age group than the 2~5-year-old age group. Furthermore, the WHTR also had less correlation with age than WC in the 6~18-year-old age group. The AUC of WHTR in identifying overweight and obesity was significantly higher than the AUC of WC in the 6~18-year-old age group. The optimal cutoff values were 0.51 in boys and 0.49 in girls for obesity, and 0.48 in boys and 0.47 in girls for overweight, with all having the AUC>0.9. The optimal cutoff values of WHTR had a higher sensitivity for diagnosing obesity than WC> or =90th percentiles. CONCLUSION: The WHTR is an easy, accurate, and less age-dependent index with high applicability in screening for obesity in children and adolescents.

Effect of dihydrotestosterone on mouse embryonic stem cells exposed to H(2)O(2)-induced oxidative stress

Oxidative stresses induced by reactive oxygen species (ROS) have been shown to be involved in several physiological and pathophysiological processes, such as cell proliferation and differentiation. Steroid hormones can protect cells against apoptosis or induce cell proliferation by several mechanisms. Among androgenic hormones, dihydrotestosterone (DHT) is generated by a 5alpha- reduction of testosterone. Unlike testosterone, DHT cannot be aromatized to estradiol, therefore DHT is considered a pure androgenic steroid. This study was conducted to examine the effect of DHT (10(-7) M) on H(2)O(2) (10(-3) M) -induced injuries in mouse embryonic stem (ES) cells. H(2)O(2) induced ROS generation and increased lipid peroxide formation and DNA fragmentation. These effects of H(2)O(2) were inhibited by pretreatment with DHT. H(2)O(2) also increased the phosphorylation of p38 MAPK, SAPK/JNK and nuclear factor kappa B (NF-kappaB), but DHT blocked these effects. Moreover, H(2)O(2) decreased DNA synthesis and the levels of cell cycle regulatory proteins [cyclin D1, cyclin E, cyclin-dependent kinase (CDK) 2, and CDK 4]. These effects of H(2)O(2) were inhibited by pretreatment with DHT. In conclusion, DHT may partially prevent H(2)O(2)-induced cell injury through inhibition of ROS and ROS-induced activation of p38 MAPK, SAPK/JNK and NF-kappaB in mouse ES cells.

Effects of Testosterone on the Expression of Transforming Growth Factor (TGF)-beta1 in Rat Penile Corpus Cavernosum / 대한남성과학회지

PURPOSE: The aim of this study was to elucidate the effect of testosterone on the expression of transforming growth factor (TGF)-beta1 in rat penile corpus cavernosum. MATERIALS AND METHODS: Male Sprague Dawley rats (n=60) were divided into 3 groups: control, castration, and testosterone replacement after castration. Testosterone was administered for 7 days at week 1 and week 2 after castration. The intracavernosal pressure/systemic blood pressure was recorded after pelvic nerve stimulation. Expression of TGF-beta1 was determined by immunoblotting and immunohistochemistry. RESULTS: At week 1 and 2 after castration, the intracavernosal pressure/systemic blood pressure was significantly decreased in the castration group (31.3+/-15.7%, 18.6+/-4.6%) compared to the control group (58.0+/-11.4%, 58.9+/-8.2%) and testosterone replacement group (50.6+/-14.5%, 49.1+/-19.4%)(p<0.05). The expression of TGF-beta1 was increased at both 1 and 2 weeks of castration compared to the control group, and was down-regulated with testosterone replacement. CONCLUSIONS: These results suggest that the testosterone maintains erectile function and regulates the expression of TGF-beta1 in the rat penile corpus cavernosum.

Effects of Testosterone on the Intracavernosally Injected Embryonic Stem Cells in Rat / 대한남성과학회지

PURPOSE: Stem cell-based cell therapy has recently been tried as a way to restore cavernosal function in an animal model. The aim of this study was to elucidate the role of testosterone on intracavernosally injected embryonic stem cells (ESCs) in rat. MATERIALS AND METHODS: Male Sprague Dawley rats(12 weeks old; n=25) were divided into five groups: control, castration, ESC injection after castration, testosterone supplementation after castration, and ESC injection with testosterone supplementation after castration(n=5 in each group). ESCs were transfected with firefly luciferase attached to adenovirus and then injected intracavernously. Testosterone propionate(0.1 mg) was subcutaneously injected daily in the testosterone supplementation group. Cell survival was assessed by optical molecular imaging the day after the ESC injection. After 4 weeks of treatment, intracavernosal pressure and systemic arterial pressure were recorded after pelvic nerve stimulation. Serum testosterone levels were measured by radioimmunoassay. RESULTS: With optical molecular imaging, we observed fluorescent signals around the external genitalia of all ESC-injected animals. The percentage of intracavernosal pressure/systolic blood pressure was significantly lower in the castration group(15.9+/-5.3%) compared to the control group(53.7+/-9.7%)(p0.05). The ESC injection with testosterone supplementation group(54.2+/-4.3%) showed erectile function similar to the control group. CONCLUSIONS: Intracavernosal injection of embryonic stem cells did not improve erectile function in the castrated rat. This result implies that testosterone may play an essential role in the proliferation of stem cells in the corpus cavernosum.

Effects of Testosterone on the Intracavernosally Injected Embryonic Stem Cells in Rat / 대한남성과학회지

PURPOSE: Stem cell-based cell therapy has recently been tried as a way to restore cavernosal function in an animal model. The aim of this study was to elucidate the role of testosterone on intracavernosally injected embryonic stem cells (ESCs) in rat. MATERIALS AND METHODS: Male Sprague Dawley rats(12 weeks old; n=25) were divided into five groups: control, castration, ESC injection after castration, testosterone supplementation after castration, and ESC injection with testosterone supplementation after castration(n=5 in each group). ESCs were transfected with firefly luciferase attached to adenovirus and then injected intracavernously. Testosterone propionate(0.1 mg) was subcutaneously injected daily in the testosterone supplementation group. Cell survival was assessed by optical molecular imaging the day after the ESC injection. After 4 weeks of treatment, intracavernosal pressure and systemic arterial pressure were recorded after pelvic nerve stimulation. Serum testosterone levels were measured by radioimmunoassay. RESULTS: With optical molecular imaging, we observed fluorescent signals around the external genitalia of all ESC-injected animals. The percentage of intracavernosal pressure/systolic blood pressure was significantly lower in the castration group(15.9+/-5.3%) compared to the control group(53.7+/-9.7%)(p0.05). The ESC injection with testosterone supplementation group(54.2+/-4.3%) showed erectile function similar to the control group. CONCLUSIONS: Intracavernosal injection of embryonic stem cells did not improve erectile function in the castrated rat. This result implies that testosterone may play an essential role in the proliferation of stem cells in the corpus cavernosum.

Effect of Korean Red Ginseng on Vaginal Blood Flow and Structure in Hypercholesterolemic Female Rats / 대한남성과학회지

PURPOSE: The aims of this study were to investigate the effects of Korean Red Ginseng(KRG) on vaginal blood flow and histological changes in a hypercholesterolemic female rat model. MATERIALS AND METHODS: Female Sprague-Dawley rats were divided into two groups: the control(n=20) and the hypercholesterolemia(n=40). Hypercholesterolemia group was fed a high fat diet(2% cholesterol, 1% cholic acid, 5% coconut oil) for 12 weeks. The hypercholesterolemia group was further divided into the vehicle only and the KRG treatment(50 mg/kg/day) groups. After 4 and 8 weeks of treatment, vaginal blood flow was measured by a laser Doppler flowmeter. Vaginal tissues were processed for histology and Western blot. RESULTS: After 4 and 8 weeks of treatment, serum cholesterol levels(mg/dl) were significantly increased in the hypercholesterolemia group(1185.0+/-736.1, 934.3+/-212.3) compared with the control group(69.7+/-19.5, 67.1+/-7.2), and partially decreased in KRG treatment group(688.2+/-251.5, 694.2+/-150.4), respectively. Vaginal blood flow(ml/min/100 g tissue) after pelvic nerve stimulation was lower in the hypercholesterolemia group(17.3+/-7.9, 17.9+/-5.5) compared with the control group(27.3+/-17.1, 26.9+/-16.4), however, the KRG treatment group(29.5+/-10.3, 27.4+/-11.1) was as high as the control group, respectively. The expressions of TGF-beta1 tended to increase in the vagina of the hypercholesterolemia animals compared to those of the control and the KRG treatment groups. CONCLUSIONS: KRG treatment in hypercholesterolemic female rats decreased serum cholesterol levels and improved vaginal blood flow. These results suggest that KRG treatment may have a beneficial effect in women's sexual health.

A Questionnaire on Using Informed Consents of Parents or Guardians in Vaccination of Children

PURPOSE: This study was done to evaluate the parents' or guardians' compliance of obtaining individual informed consent before vaccination and the possible problems of using informed consent. METHODS: One hundred and twelve children were randomly selected among 2,820 children who visited Samsung Cheil Hospital during July 2002. A questionnaire about using informed consent of vaccination was given to all parents or guardians of these children. RESULTS: Most of the responders were mothers(80.4%). As for the places of vaccination, 87(77.7%) persons vaccinated their children at general hospitals. In response to a request for informed consent of vaccination, 73(65.2%) complied well with requests every time. In subscribing to informed consent, 62(55.4%) were pleased, 40(35.7%) stated it was necessary despite in inconveniance. Because of dislike of subscribing to informed consent(55), 33(60%) persons stated that it was difficult to subscribe to informed consent during the inconveniant time of nursing baby. Eighty(71.4%) found it useful to read informed consent for an understanding of potential negative reactions. Being informed of potential negative reactions, 105(93.8%) stated that they would vaccinate their children despite of the potential of a worrisome negative reaction. In dealing with negative reaction, 77(68.8%) said that they would contact a hospital, 32(28.6%) said that they would care for their children at home. Eleven(9.8 %) experienced negative reactions after vaccination. CONCLUSION: Compliance of parents to informed consent was relatively good. Parents or guardians should receive relevant information and the aim of informed consent also should be adequately understood.