• Title/Summary/Keyword: Tissue specificity

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Development of Enzymatic Recombinase Amplification Assays for the Rapid Visual Detection of HPV16/18

  • Ning Ding;Wanwan Qi;Zihan Wu;Yaqin Zhang;Ruowei Xu;Qiannan Lin;Jin Zhu;Huilin Zhang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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    • pp.1091-1100
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    • 2023
  • Human papillomavirus (HPV) types 16 and 18 are the major causes of cervical lesions and are associated with 71% of cervical cancer cases globally. However, public health infrastructures to support cervical cancer screening may be unavailable to women in low-resource areas. Therefore, sensitive, convenient, and cost-efficient diagnostic methods are required for the detection of HPV16/18. Here, we designed two novel methods, real-time ERA and ERA-LFD, based on enzymatic recombinase amplification (ERA) for quick point-of-care identification of the HPV E6/E7 genes. The entire detection process could be completed within 25 min at a constant low temperature (35-43℃), and the results of the combined methods could be present as the amplification curves or the bands presented on dipsticks and directly interpreted with the naked eye. The ERA assays evaluated using standard plasmids carrying the E6/E7 genes and clinical samples exhibited excellent specificity, as no cross-reaction with other common HPV types was observed. The detection limits of our ERA assays were 100 and 101 copies/µl for HPV16 and 18 respectively, which were comparable to those of the real-time PCR assay. Assessment of the clinical performance of the ERA assays using 114 cervical tissue samples demonstrated that they are highly consistent with real-time PCR, the gold standard for HPV detection. This study demonstrated that ERA-based assays possess excellent sensitivity, specificity, and repeatability for HPV16 and HPV18 detection with great potential to become robust diagnostic tools in local hospitals and field studies.

Cytokeratin 15 is an Effective Indicator for Progression and Malignancy of Esophageal Squamous Cell Carcinomas

  • Shen, Yu-Hong;Xu, Cui-Ping;Shi, Zhi-Meng;Zhang, Yan-Jiao;Qiao, Ya-Guang;Zhao, He-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4217-4222
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    • 2016
  • Purpose: To compare the expression level of CK 15 in normal esophageal and esophageal squamous-cell carcinoma (ESCC) tissues and analyse possible functions of CK15 in occurrence and development. Materials and Methods: Immunohistochemistry was used to compare CK14, CK15 and proliferating cell nuclear antigen (PCNA) expression levels in ESCCs. Expression level of CK15 was also assessed by Western blotting. In addition, levels of CK15, cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) and PCNA were detected in serum by enzymelinked immunosorbent assay (ELISA) and chemiluminescence methods. Relationships between clinicopathological parameters and CK14 and CK15 expression were then analyzed. Results: According to immunohistochemistry, in esophageal and intraepithelial neoplasia (SIN) tissues, the expression of CK14, CK15 and PCNA localized to basal layer of the epithelium. CK14 and CK15 levels were higher in normal esophageal squamous epithelial tissue than in SIN and ESCC, and greater in highly differentiated than poorly differentiated carcinoma tissue. By Western blotting, we found more pronounced expression of CK15 in normal esophageal tissue, compared with carcinoma tissue. The specificity of changed CK15 and CYFRA21-1 expression was respectively 90.0% and 96.7% in serum of ESCC patients. Joint detection could improve the sensitivity of esophageal carcinoma diagnosis. Relationships between CK14, CK15 expression and clinical parameters were not statistically significant (P>0.05). Postoperative survival in patients of CK14, CK15 positive expression was longer than with negative expression ($x^2=4.35$, P=0.037; $x^2=9.852$, P=0.002). Conclusions: CK15 expression decreased in esophageal squamous cell carcinoma tissue and serum of esophageal squamous carcinoma patients. We infer that CK15 may play an important role for the occurrence and development of esophageal squamous-cell carcinoma. In the future, CK15 may be used for the diagnosis, treatment and prognostic evaluation of esophageal squamous-cell carcinoma.

Clinical Usefulness of PET-MRI in Lymph Node Metastasis Evaluation of Head and Neck Cancer (두경부암 림프절 전이 평가에서 PET-MRI의 임상적 유용성)

  • Kim, Jung-Soo;Lee, Hong-Jae;Kim, Jin-Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.26-32
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    • 2014
  • Purpose: As PET-MRI which has excellent soft tissue contrast is developed as integration system, many researches about clinical application are being conducted by comparing with existing display equipments. Because PET-MRI is actively used for head and neck cancer diagnosis in our hospital, lymph node metastasis before the patient's surgery was diagnosed and clinical usefulness of head and neck cancer PET-MRI scan was evaluated using pathological opinions and idiopathy surrounding tissue metastasis evaluation method. Materials and Methods: Targeting 100 head and neck cancer patients in SNUH from January to August in 2013. $^{18}F-FDG$ (5.18 MBq/kg) was intravenous injected and after 60 min of rest, torso (body TIM coil, Vibe-Dixon) and dedication (head-neck TIM coil, UTE, Dotarem injection) scans were conducted using $Bio-graph^{TM}$ mMR 3T (SIEMENS, Munich). Data were reorganized using iterative reconstruction and lymph node metastasis was read with Syngo.Via workstation. Subsequently, pathological observations and diagnosis before-and-after surgery were examined with integrated medical information system (EMR, best-care) in SNUH. Patient's diagnostic information was entered in each category of $2{\times}2$ decision matrix and was classified into true positive (TP), true negative (TN), false positive (FP) and false negative (FN). Based on these classified test results, sensitivity, specificity, accuracy, false negative and false positive rate were calculated. Results: In PET-MRI scan results of head and neck cancer patients, positive and negative cases of lymph node metastasis were 49 and 51 cases respectively and positive and negative lymph node metastasis through before-and-after surgery pathological results were 46 and 54 cases respectively. In both tests, TP which received positive lymph node metastasis were analyzed as 34 cases, FP which received positive lymph node metastasis in PET-MRI scan but received negative lymph node metastasis in pathological test were 4 cases, FN which received negative lymph node metastasis but received positive lymph node metastasis in pathological test was 1 case, and TN which received negative lymph node metastasis in both two tests were 50 cases. Based on these data, sensitivity in PET-MRI scan of head and neck cancer patient was identified to be 97.8%, specificity was 92.5%, accuracy was 95%, FN rate was 2.1% and FP rate was 7.00% respectively. Conclusion: PET-MRI which can apply the acquired functional information using high tissue contrast and various sequences was considered to be useful in determining the weapons before-and-after surgery in head and neck cancer diagnosis or in the evaluation of recurrence and remote detection of metastasis and uncertain idiopathy cervical lymph node metastasis. Additionally, clinical usefulness of PET-MRI through pathological test and integrated diagnosis and follow-up scan was considered to be sufficient as a standard diagnosis scan of head and neck cancer, and additional researches about the development of optimum MR sequence and clinical application are required.

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A methodological study on simplifying claims review system in medical insurance (의료보험 진료비 심사 간소화에 대한 방법론적 연구)

  • Kim, Suk-Il;Kang, Hyung-Gon;Kim, Han-Joong;Chae, Young-Moon;Sohn, Myong-Sei;Lee, Myung-Keun
    • Journal of Preventive Medicine and Public Health
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    • v.28 no.3 s.51
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    • pp.640-650
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    • 1995
  • After the introduction of National Medical Insurance in 1989, the medical demand has rapidly increased. The impact of increased medical demand was followed by an increase in the number of claims in need of review. We studied a new, fair method for reducing the number of claims reviewed. We analysed 90,583 outpatient claims submitted between September and October; claims were made for services given August of 1994. We finally suggested a screening system for claims review using a statistical method of discriminant analysis of the medical costs. The results were as follows. 1. In the cut-off group, age, days of medication, number of hospital or clinic visits, and total charge were significantly high. The cut-off rates according to the hospital-type and existence of accompanied disease were significantly different 2. According to ICD, the cut-off rate was highest in peripheral enthesopathies and allied syndromes(20.76%), lowest in acute sinusitis(0.93%). The mean charges were significantly different according to ICD and existence of cut-off. 3. We build discriminant functions by ICD with such discriminant variables as patient age, sex, existence of accompanied disease, number of hospital or clinic visits, and 9 detailed hospital or clinic charges included in claim. 4. We applied the discriminant function for screening those claims that were expected to be cut-off. The sensitivities comprised from 40% to 70%, and specificities from 70% to 95% by ICD. Acute rhinitis had highest sensitivity(100.00%) and other local infections of skin and subcutaneous tissue had highest specificity(98.45%). The expected number of cut-off was 17,762(19.61%). The total sensitivity was 49.62%, the total specificity was 82.57% and the error rate was 19.66%. We lacked economic analysis such as cost-benefit analysis. But, if the new method of screening claims using discriminant analysis were applied, the number of claims in need of review will reduce considerably.

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Importance of pulmonary valve morphology for pulmonary valve preservation in tetralogy of Fallot surgery: comparison of the echocardiographic parameters

  • Choi, Su Jin;Kwon, Jung Eun;Roh, Da Eun;Hyun, Myung Chul;Jung, Han Na;Lee, Young Ok;Cho, Joon Yong;Kim, Yeo Hyang
    • Clinical and Experimental Pediatrics
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    • v.63 no.5
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    • pp.189-194
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    • 2020
  • Background: The decision to use transannular patching (TAP) during tetralogy of Fallot (TOF) repair depends on the pulmonary valve annulus size; the z score of the pulmonary annulus is the most commonly used predictor. However, definitive results are not obtained with z scores as different z score data sets are used for different parameters. Purpose: This study aimed to identify the echocardiographic and other key factors that warranted a change in the surgical method during TOF surgery. Methods: Sixty-two patients were enrolled and divided into a pulmonary valve (PV) preservation group and a TAP group. Their medical records were reviewed. Results: The z score for PV annulus (PVA), ratio of the PVA to aortic annulus size, and ratio of PVA to descending aorta (DAO) size were significantly different between the PV preservation and TAP groups (-1.72±1.52 vs. -3.07±1.94, P=0.004; 0.62±0.12 vs. 0.50±0.14, P=0.002; and 1.32±0.32 vs. 1.07±0.36, P=0.008, respectively). For TAP repair, the PVA z score had a sensitivity of 65.4% and specificity of 73.1%, ratio of PVA to aortic annulus size had a sensitivity of 73.1% and specificity of 65.4%, and ratio of PVA to DAO size had a sensitivity of 69.2% and specificity of 57.7%. The TAP group showed more monocuspid PVs (P=0.011), while the PV preservation group showed more tricuspid PVs (P=0.027). Commissurotomy was more frequently performed in the PV preservation group than in the TAP group (P=0.001). Of patients with commissurotomy, 58% showed a PV z score<-2. Conclusion: Although various echocardiographic parameters may serve as predictors for determining surgical methods for TOF patients, the PV morphology and tissue characteristics should also be considered.

Ultrasounds Image on the Disorders of the Ligaments Surrounding Temporomandibular Joints (측두하악관절 주변인대 질환의 초음파영상)

  • Hong, Soo-Min;Im, Yeong-Gwan;Kim, Byung-Gook
    • Journal of Oral Medicine and Pain
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    • v.33 no.4
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    • pp.387-394
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    • 2008
  • Purpose : The purpose of this study was to verify the usefulness and feasibility of ultrasonographic imaging for the detection of the disorders of the surrounding supporting structure such as articular capsule, retrodiscal tissue and related ligaments, osteoarthritic evidence and associated disc displacement at the temporomandibular joint(TMJ) Materials and Methods : 20 patients(40 joints) with periodic lock and crepitations were investigated prospectively using 12 MHz array transducer. Ultrasonographic Imagings were assessed for osteoarthritic surface changes of condyle, extent of disc displacement and disorders of surrounding structures. Ultrasonographic images were compared with clinical investigations, conventional radiography and Dental Computed Tomographic scans. Results : In clinical and conventional radiography, osteoarthritic changes were diagnosed in 8 joints. Ultrasonographically 7 of the 8 osteoarthritic changes were diagnosed correctly. Sensitivity, specificity, and accuracy of ultrasonography in the osteoarthritic detections were 87.5%, 62.5%, and 67.5% respectively. About the detection of disc displacement, disc displacement were diagnosed in 21 joints clinically. Ultrasonographically 19 of the 21 disc displacements were diagnosed correctly. Sensitivity, specificity, and accuracy of ultrasonography in the osteoarthritic detections were 95%, 90%, and 92.5% respectively. when the disorders of supportive structure were figured out, the disorders of supportive structure were diagnosed in 18 joints clinically. Ultrasonographically 1 of the 18 the disorders of supportive structure were diagnosed correctly. Sensitivity, specificity, and accuracy of ultrasonography in the osteoarthritic detections were 5.5%, 4.5%, and 55% respectively. Conclusion : Ultrasonography is an relatively reliable diagnostic tool for the detection of disc displacement and some of osteoarthritic changes. But it's not an insufficient imaging technique for the detection of the disorders of the surrounding structure.

MRI Evaluation of Suspected Pathologic Fracture at the Extremities from Metastasis: Diagnostic Value of Added Diffusion-Weighted Imaging

  • Sun-Young Park;Min Hee Lee;Ji Young Jeon;Hye Won Chung;Sang Hoon Lee;Myung Jin Shin
    • Korean Journal of Radiology
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    • v.20 no.5
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    • pp.812-822
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    • 2019
  • Objective: To assess the diagnostic value of combining diffusion-weighted imaging (DWI) with conventional magnetic resonance imaging (MRI) for differentiating between pathologic and traumatic fractures at extremities from metastasis. Materials and Methods: Institutional Review Board approved this retrospective study and informed consent was waived. This study included 49 patients each with pathologic and traumatic fractures at extremities. The patients underwent conventional MRI combined with DWI. For qualitative analysis, two radiologists (R1 and R2) independently reviewed three imaging sets with a crossover design using a 5-point scale and a 3-scale confidence level: DWI plus non-enhanced MRI (NEMR; DW set), NEMR plus contrast-enhanced fat-saturated T1-weighted imaging (CEFST1; CE set), and DWI plus NEMR plus CEFST1 (combined set). McNemar's test was used to compare the diagnostic performances among three sets and perform subgroup analyses (single vs. multiple bone abnormality, absence/presence of extra-osseous mass, and bone enhancement at fracture margin). Results: Compared to the CE set, the combined set showed improved diagnostic accuracy (R1, 84.7 vs. 95.9%; R2, 91.8 vs. 95.9%, p < 0.05) and specificity (R1, 71.4% vs. 93.9%, p < 0.005; R2, 85.7% vs. 98%, p = 0.07), with no difference in sensitivities (p > 0.05). In cases of absent extra-osseous soft tissue mass and present fracture site enhancement, the combined set showed improved accuracy (R1, 82.9-84.4% vs. 95.6-96.3%, p < 0.05; R2, 90.2-91.1% vs. 95.1-95.6%, p < 0.05) and specificity (R1, 68.3-72.9% vs. 92.7-95.8%, p < 0.005; R2, 83.0-85.4% vs. 97.6-98.0%, p = 0.07). Conclusion: Combining DWI with conventional MRI improved the diagnostic accuracy and specificity while retaining sensitivity for differentiating between pathologic and traumatic fractures from metastasis at extremities.

Usefulness of Automated PCR Test for the Detection of Mycobacterium Tuberculosis in Fresh Biopsy Tissues (신선조직 검체에서 결핵균 검출을 위한 자동화 중합효소연쇄반응 검사의 유용성)

  • Choi, Woo Soon;Shin, So Young;Kim, Jong Ok;Kim, Myung Sook;Lee, Hye Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.1
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    • pp.54-59
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    • 2006
  • Background: Although there have been several studies regarding the clinical value of an automated TB-PCR study using sputum, bronchial washing, and other body fluid samples for the detection of pulmonary tuberculosis, there are only a few reports on the use of fresh tissue samples. Materials and methods: The acid-fast bacilli stain(AFB), tuberculosis culture, automated TB-PCR study, and histopathology examination were performed in 42 fresh tissue samples. Results: Among the 42 cases, 18 cases were diagnosed with tuberculosis based on the clinical findings. Sixteen of the 18 cases were TB-PCR positive and of these 16 cases, only 2 cases were positive in the AFB stain or culture study. However, all 18 cases showed the histopathology findings of chronic granulomatous inflammation that was compatible with tuberculosis. Based on the clinical findings, the sensitivity, specificity, positive predictability, and negative predictability of the automated TB-PCR study were 88.9%, 100%, 100%, and 92.3% respectively. Conclusion: An automated TB-PCR assay is an important diagnostic tool for diagnosing tuberculosis in fresh tissue samples.

Effects of an in vitro vitamin D treatment on the inflammatory responses in visceral adipose tissue from Ldlr-/- mice

  • Deok Hoon Kwon;Jungwon Hwang;Hyeyoung You;Na Young Kim;Ga Young Lee;Sung Nim Han
    • Nutrition Research and Practice
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    • v.18 no.1
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    • pp.19-32
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    • 2024
  • BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlrtm1Her/J (Ldlr-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)2D3 for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)2D3 treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)2D3 treatment. The 1,25(OH)2D3 treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

Analysis of Observer Agreement in Shear Wave Elastography using a Breast Phantom (유방 팬텀을 활용한 전단파 탄성초음파 검사에서의 관측자 일치도 분석)

  • Jin-Hee Kim;Jung-Hoon Kim;Sung-Hee Yang
    • Journal of the Korean Society of Radiology
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    • v.18 no.4
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    • pp.355-363
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    • 2024
  • This study evaluated the clinical utility of Shear Wave Elastography(SWE) by analyzing the differences in elastic modulus and shear wave speed across various types of breast tissue and assessing inter-observer agreement. A breast phantom that included normal breast tissue, benign tumors, and malignant tumors was utilized, and ten radiologists participated, measuring the minimum, average, and maximum elastic modulus and shear wave speed for each tissue type. Analysis of differences between tissues was conducted using one-way ANOVA, and intra- and inter-observer agreement was assessed using the Intraclass Correlation Coefficient(ICC). The results demonstrated significant differences in the average values of elastic modulus and shear wave speed among the tissue types(p<0.001), with malignant tumor tissues showing the highest average values. Furthermore, the ICC analysis for elastic modulus ranged from 0.75 to 0.99 and for shear wave speed from 0.89 to 0.99, indicating high reproducibility and agreement. These findings suggest that SWE is a reliable tool with high reproducibility and specificity for the diagnosis of breast cancer.