Recently, next generation sequencing (NGS) has received attention as the ultimate genotyping method to overcome the limitations of capillary electrophoresis (CE)-based short tandem repeat (STR) analysis, such as the limited number of STR loci that can be measured simultaneously using fluorescent-labeled primers and the maximum size of STR amplicons. In this study, we analyzed 15 autosomal STR markers via the NGS method and evaluated their effectiveness in STR analysis. Using male and female standard DNA as single-sources and their 1:1 mixture, we sequentially generated sample amplicons by the multiplex polymerase chain reaction (PCR) method, constructed DNA libraries by ligation of adapters with a multiplex identifier (MID), and sequenced DNA using the Roche GS Junior Platform. Sequencing data for each sample were analyzed via alignment with pre-built reference sequences. Most STR alleles could be determined by applying a coverage threshold of 20% for the two single-sources and 10% for the 1:1 mixture. The structure of the STR in each allele was accurately determined by examining the sequences of the target STR region. The mixture ratio of the mixed sample was estimated by analyzing the coverage ratios between assigned alleles at each locus and the reference/variant ratios from the observed sequence variations. In conclusion, the experimental method used in this study allowed the successful generation of NGS data. In addition, the NGS data analysis protocol enables accurate STR allele call and repeat structure determination at each locus. Therefore, this approach using the NGS system will be helpful to interpret and analysis the STR profiles from singe-source and even mixed samples in forensic investigation.
Alleles ; DNA ; Electrophoresis, Capillary ; Female ; Gene Library ; Humans ; Ligation ; Male ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction ; Statistics as Topic

Human mitochondrial DNA (mtDNA) is generally used to identify highly degraded forensic samples, particularly when the extracted DNA is not sufficient for nuclear DNA analysis. However, direct sequencing, the most widely used mtDNA analysis method, is laborious and time-consuming, and precludes the simultaneous analysis of many samples. Here, we describe a rapid and simple screening method for mtDNA analysis in Koreans using single base extension (SBE) methods. Sixteen highly polymorphic mtDNA SNPs from the control region were selected, and a multiplex SBE system was constructed to analyze them. Because the developed system consists of two duplex PCRs, which produce small amplicons with fewer than 270 bp, it works well with highly degraded samples such as old skeletal remains. Using this multiplex SBE system, 145 different haplotypes were expected to be observed from 593 unrelated Koreans. Seventy-three haplotypes were expected to be observed only once, and the most frequent haplotype was expected to occur 80 times. Since the mean number of pairwise differences was estimated to be 4.55, the developed system could be useful to exclude samples that do not match evidence and reference samples. Therefore, the multiplex SBE system used in this study will be a useful tool to analyze many samples simultaneously and to efficiently screen out non-matching mtDNA sequences in forensic casework.
Asian Continental Ancestry Group ; DNA ; DNA, Mitochondrial* ; Haplotypes ; Humans ; Mass Screening* ; Methods ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide*

No abstract available.
Beckwith-Wiedemann Syndrome*

This experiment was performed to investigate the response of inorganic substances in alveolar bone in relation to the experimental tooth movement. Right canine in maxillary jaw was tipped in cats by coil springs generating 80 gm. force, in mandibular jaw, the force was 100 gm. force. Cats were divided into five groups and orthodontically treated for one hour, 1, 7, 14 and 28 days, respectively. Alveolar bone samples were obtained from tension and compression sites as well as from contralateral control sites. The level of calcium of alveolar bone was determined by atomic absorption spectrophotometry and inorganic phosphorus was measured by spectrophotometry. The results obtained were-as follows: 1. In tension and compression site of maxillary alveolar bone, calcium levels were decreased at 1, 7 and 14 days, but recovered at 28 days. 2. The levels of inorganic phosphorus in compression site of maxillary alveolar bone had little change but in tension site of maxillary alveolar bone,, phosphorous levels were decreased. 3. Calcium levels in tension and compression site of mandibular alveolar bone were decreased, especially at 28 days. 4. In tension and compression site of mandibular alveolar bone, inorganic phosphorus were slightly decreased from I day.
Animals ; Calcium* ; Cats* ; Jaw ; Phosphorus ; Spectrophotometry ; Spectrophotometry, Atomic ; Tooth Movement

Cervical cancer and its precursors are caused principally, if not exclusively, by HPV infection and HPV DNA is found in more than 90% of cervical cancers. Cervical cytology is limited by its false negativity and this may be supplimented by other adjunctive test such as HPV test. It is therefore important to explore the use of HPV DNA detection as a primary or supplementary screening method and to determine whether HPV typing can be used as a predictor of a lesion's clinical behavior. Cervical cytology and Hybrid Capture test for HPV detection were performed in 450 asymtomatic wornen visited Health Care Center in Kangnam St. Mary's Hospital,and none of whom was believed to have current cervical disease. The Papanicolaou cytology results were classified by The Bethesda System : 333(74.%) women were classified to within normal limit, 19(4.2%) benign reactive change, 38(8.4%) ASCUS, 59(13%) low grade SIL, and only one woman high grade SIL. Twenty five of 450(5.6%) women showed HPV infection by Hybid Capture test. Among 98 wornen with abnormal Papanicolaou cytology, 16(16.3%) women showed HPV DNA positivity. (continue)
Delivery of Health Care ; DNA* ; Female ; Humans* ; Mass Screening* ; Papilloma* ; Uterine Cervical Neoplasms

PURPOSE: The aim of this study is to evaluate the sequential histologic changes and spermatogenesis of the testis after extracorporeal shock wave treatment (ESWT). MATERIALS AND METHODS: ESWT to right testis was applied on total 48 immature, 5-week-old Sprague Dawley rats. The rats were sacrficed immediately, at 1 day, 3 days, 1 week, 2 weeks and 4 weeks after ESWT, and each group consisted of 8 rats including 2 rats as control (without ESWT). Histologic changes of both testes were examined, and serial three or more parafin sections were obtained, and immunohistochemical stains for bcl-2 and Hsp 70 and TUNEL method for apoptosis were performed. RESULTS: Hemorrhage and destruction of seminiferous tubules which appeared immediately after ESWT were maximal at 1 day after ESWT and were seen absorption and atrophy at 1 week after ESWT. Even regenerating tubules were found at 2 weeks after ESWT, no significant histologic difference between both testes experimental and control groups were seen except for some regenerating tubules. The right testicular tissues, remote from the ESWT-targeted area, showed preserved spermatogenesis since the age of 5 weeks 3 days, and was presented no significant difference between experimental and control groups. Hsp 70 and bcl-2 expression and apoptosis were maximal at 1 day after ESWT and persisted to 3 day after ESWT. Weak and focal expression were noted at 1 week after ESWT, and mostly disappeared at 2 weeks after. CONCLUSIONS: Spermatogenesis of right testis except the targeted area was not diffrent compared with that of left testis and control group. Hsp-70 expressin, which limits tissue destruction after injury, was immediately appeared for defense mechanism, and hemorrhage and tissue destruction following ESWT seemed to be due to apoptosis controlled by bcl-2 expression.
Absorption ; Animals ; Apoptosis* ; Atrophy ; Coloring Agents ; Hemorrhage ; In Situ Nick-End Labeling ; Rats* ; Rats, Sprague-Dawley ; Seminiferous Tubules ; Shock* ; Spermatogenesis ; Testis*

Wegener's granulomatosis is characterized by necrotizing granulomatosis lesion of the respiratory tract, glomerulonephritis and frequently vasculitis involving other organs. The basic pathophysiologic mechanism of Wegener's granulomatosis is not defined yet. However, it may be suspected an autoimmune disease. We experienced a case of Wegener's granulomatosis which are associated with acute renal failure and pneumothorax. The patient suffered from hemoptysis, fever and cough. Despite antibiotic therapy, symtoms did not improved and multiple varying sized nodules were aggravated on chest roentogenogram and serum creatinine elevated 3.4mg/dl. After diagnosis using video associated thoracoscopic surgery, the patient was treated with cyclophsphamide, glucocorticoid and sulfamethoxasole-trimethoprime. With the combination therapy, the patient felt completely well and chest roentogenogram showed lungs were improved and serum creatinine was normal. The patient was readmitted because of right pleuritic pain and dyspnea 15 day after discharge. The patient was developed a right pneumothorax. The lung was easily expanded by intercostal tube drainage with a one way valve. The patient has been treated as an out- patient with immunosuppressive agents continously.
Acute Kidney Injury* ; Autoimmune Diseases ; Cough ; Creatinine ; Diagnosis ; Drainage ; Dyspnea ; Fever ; Glomerulonephritis ; Hemoptysis ; Humans ; Immunosuppressive Agents ; Lung ; Pneumothorax* ; Respiratory System ; Thoracoscopy ; Thorax ; Vasculitis ; Wegener Granulomatosis*

Short stature is common in case of Fanconi anemia (FA) and may result from inherent abnormalities of FA or endocrinopathies, such as a growth hormone deficiency and hypothyroidism. Growth hormone treatment in FA patients with growth hormone deficiency (GHD) has been controversial because FA itself has a risk of malignancy, and there has been a concern about the increased risk of malignancy from growth hormone treatment. We report on an FA boy with GHD who was diagnosed with FA and undertook stem cell transplantation. Because he showed severe growth failure, growth hormone therapy was initiated at 6 years of age, after he was diagnosed with hypopituitarism, including GHD. He expressed significant catch-up growth with no complications beyond the two year follow up period. This case suggests that growth hormone therapy can be considered in patients with GHD caused by FA, with careful monitoring for malignancy.
Fanconi Anemia ; Follow-Up Studies ; Growth Hormone ; Humans ; Hypopituitarism ; Hypothyroidism ; Stem Cell Transplantation

OBJECTIVE: This study evaluates the effect of the specific human papillomavirus (HPV) genotype as a prognostic factor in stage I-IIA cervical cancer patients following primary surgical treatment. METHODS: The medical records of 116 cervical cancer patients treated with primary surgical treatment were reviewed. The HPV genotypes were categorized into following groups: negative and unclassified, HPV 16, HPV 18, and other high risk (HPV 31, 33, 35, 45, 51, 52, 56, and 58). RESULTS: Among the HPV genotypes, HPV 16 predominated (40.52%), followed by intermediate risk and unclassified (25%), HPV 18, 45, and 56 (17.24%) and negative (17.24%). In univariate analysis, HPV genotypes (P=0.03), parametrial spread (P=0.02), depth of invasion (DOI) (P<0.01) and lymph-vascular space invasion (P=0.02) were significantly associated with progression free survival (PFS). In multivariate analysis, HPV 18 (hazard ratio [HR], 5.2; 95% confidence interval [CI], 1.29 to 20.90; P=0.02) and > or =one half of DOI (HR, 5.4; 95% CI, 1.08 to 27.31; P=0.04) were significantly associated with PFS. HPV genotypes are not significantly associated with overall survival. CONCLUSION: HPV 18 was a poor prognostic factor for the PFS in stage I-IIA cervical cancer patients following primary surgical treatment. Careful long-term observation and regular exams are recommended for cervical cancer patients with HPV 18 compared to those with other HPV genotypes.
Disease-Free Survival ; Genotype ; Human papillomavirus 16 ; Human papillomavirus 18* ; Humans ; Medical Records ; Multivariate Analysis ; Prognosis ; Uterine Cervical Neoplasms*

BACKGROUND: Medication adherence is important for hypertension management but still stay low level. It is reasonable method to classify medication nonadherence into intentional nonadherence and unintentional nonadherence and manage it according to this categories. This study aimed to explore medication adherence and its predictors in community patients with hypertension, especially dividing into intentional nonadherence and unintentional nonadherence. METHODS: Study subjects included 1,988 patients who were prescribed hypertension drugs among 2012 community health survey subjects of 10 cities in Gyeongsangnamdo and we analyzed medication adherence with hypertension and its predictors. We conducted chi-square test for nominal variable and ANOVA test for continuous variable and use multinominal regression to analyze independent predictors of intentional nonadherence and unintentional nonadherence in contrast to medication adherence. RESULTS: Of the 1,988 patients, 49.7% were adherent, 26.1% were unintentionally nonadherent and 24.2% intentionally nonadherent. Independent predictors of unintentional nonadherence were depressive symptoms (odds ratio [OR]=1.696, P=0.047) and arthritis (OR=1.319, P=0.030) and independent predictors of intentional nonadherence were cardiocerebrovascular disease (OR=1.464, P=0.044), self-efficacy (OR=0.984, P=0.007), beliefs about medications questionnaire (necessity [OR=0.834, P<0.001] and concern [OR=1.236, P<0.001]). CONCLUSIONS: In order to manage hypertension in community, improvement in medication adherence is needed. Depressive symptom and self-efficacy need to be managed, but especially patients'beliefs about their medication need to be considered to improve intentional nonadherence.
Aged* ; Arthritis ; Depression ; Gyeongsangnam-do ; Health Surveys ; Humans ; Hypertension* ; Intention ; Medication Adherence*