ObjectiveDetection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability, health, and metabolic rate.After exposure to environmental stimuli, both the internal reference gene and target gene would be degraded. As a result, it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation. This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment. MethodsThe new RNA was labeled with 5-ethyluridine (5-EU) instead of uracil, and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of “Click Chemistry” and magnetic bead screening. Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon (M.B.) and quantitative reverse transcription PCR (qRT-PCR). ResultsThe bacterial nascent RNA captured by “Click Chemistry” screening can be used as a reverse transcription template to form cDNA. Combined with the fluorescent molecular beacon M.B.1, the synthesis rate of rRNA at 37℃ is 1.2 times higher than that at 15℃. The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃ and 16℃ when analyzed with nascent RNA rather than total RNA, enabling accurate detection of RNA transcription rates. ConclusionCompared to other article reported experimental methods that utilize screening magnetic columns, the technical scheme employed in this study is more suitable for bacteria, and the operation steps are simple and easy to implement, making it an effective RNA capture method for researchers.

Evidence of the ethical appropriateness and clinical benefits of shared decision-making (SDM) are accumulating. This study aimed to not only identify physicians’ perspectives on SDM, and practices related to end-of-life care in particular, but also to gauge the effect of SDM education on physicians in Korea. Methods: A 14-item questionnaire survey using a modified Delphi process was delivered to nephrologists and internal medicine trainees at 17 university hospitals. Results: A total of 309 physicians completed the survey. Although respondents reported that 69.9% of their practical decisions were made using SDM, 59.9% reported that it is not being applied appropriately. Only 12.3% of respondents had received education on SDM as part of their training. The main obstacles to appropriate SDM were identified as lack of time (46.0%), educational materials and tools (29.4%), and education on SDM (24.3%). Although only a few respondents had received training on SDM, the proportion of those who thought they were using SDM appropriately in actual practice was high; the proportion of those who chose lack of time and education as factors that hindered the proper application of SDM was low. Conclusion: The majority of respondents believed that SDM was not being implemented properly in Korea, despite its use in actual practice. To improve the effectiveness of SDM in the Korean medical system, appropriate training programs and supplemental policies that guarantee sufficient application time are required.

To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.

Synovial chondromatosis is a rare lesion in the wrist, but some cases in the distal radioulnar joint have been reported and previous case reports emphasize joint calcifications, shown on preoperative plain radiographs. We report an extremely uncommon case of synovial chondromatosis in the pisotriquetral joint, in which radiographs and magnetic resonance imaging did not demonstrate apparent calcified bodies. In our case, for the accurate diagnosis and treatment, surgical exploration of the joint and synovectomy with removal of loose bodies was performed.
*Chondromatosis, Synovial/diagnosis/physiopathology/surgery ; Female ; Humans ; Middle Aged ; *Wrist Joint/physiopathology/surgery

Hepatic ischemia/reperfusion (I/R) injury leads to oxidative stress and acute inflammatory responses that cause liver damage and have a considerable impact on the postoperative outcome. Much research has been performed to develop possible protective techniques. We aimed to investigate the efficacy of SPA0355, a synthetic thiourea analog, in an animal model of hepatic I/R injury. Male C57BL/6 mice underwent normothermic partial liver ischemia for 45 min followed by varying periods of reperfusion. The animals were divided into three groups: sham operated, I/R and SPA0355 pretreated. Pretreatment with SPA0355 protected against hepatic I/R injury, as indicated by the decreased levels of serum aminotransferase and reduced parenchymal necrosis and apoptosis. Liver synthetic function was also restored by SPA0355 as reflected by the prolonged prothrombin time. To gain insight into the mechanism involved in this protection, we measured the activity of nuclear factor-kappaB (NF-kappaB), which revealed that SPA0355 suppressed the nuclear translocation and DNA binding of NF-kappaB subunits. Concomitantly, the expression of NF-kappaB target genes such as IL-1beta, IL-6, TNF-alpha and iNOS was significantly downregulated. Lastly, the liver antioxidant enzymes superoxide dismutase, catalase and glutathione were upregulated by SPA0355 treatment, which correlated with the reduction in serum malondialdehyde. Our results suggest that SPA0355 pretreatment prior to I/R injury could be an effective method to reduce liver damage.
Animals ; Anti-Inflammatory Agents/*therapeutic use ; Benzoxazines/*therapeutic use ; Liver/*drug effects/immunology/*injuries/pathology ; Male ; Mice, Inbred C57BL ; NF-kappa B/immunology ; Reperfusion Injury/*drug therapy/immunology/pathology ; Signal Transduction/drug effects ; Thiourea/*analogs & derivatives/therapeutic use

BACKGROUND/AIMS: We investigated the incidence and clinical characteristics of renal cell carcinoma (RCC) in the native kidney of renal transplant recipients. METHODS: Between 1991 and 2010, 1,425 patients underwent kidney transplantation at our institution. We retrospectively evaluated the clinical features and outcomes in renal transplant patients with RCC in the native kidney after renal transplantation. RESULTS: The patients included three males and two females with a mean age of 63 years (range, 52 to 74). The incidence of RCC was 0.35%. The median interval between renal transplantation and RCC occurrence was 16.2 years (range, 9 to 20). All of our patients with RCC had developed renal cysts either before (n = 3) or after (n = 2) renal transplantation. The mean duration of dialysis was 12 months (range, 2 to 39). Of the five patients, four underwent dialysis treatment for less than 8 months. All the RCCs were low grade at the time of diagnosis. Four patients underwent radical nephrectomy, and one patient refused the operation. The four patients who underwent radical nephrectomy showed no evidence of local recurrence or distant metastasis during the median follow-up of 2.9 years. However, the patient who did not undergo surgery developed spinal metastasis from the RCC 6 years later. CONCLUSIONS: This study suggests that the follow-up period is an important factor for the development of RCC in renal transplant recipients, and more vigorous screening with a longer follow-up period is required in renal transplant recipients.
Aged ; Carcinoma, Renal Cell/*epidemiology ; Female ; Humans ; Incidence ; Kidney Neoplasms/*epidemiology ; *Kidney Transplantation ; Male ; Middle Aged ; Postoperative Complications/*epidemiology ; Republic of Korea/epidemiology ; Retrospective Studies

Since the release of first Korean Clinical Practice Guideline of Stroke in 2009, many important new evidences have emerged in the field of thrombolytic therapy. Among the recent developments are the extended therapeutic time window of intravenous (IV) tissue plasminogen activator (tPA) up to 4.5 hours after onset, and the efforts for the wider application of IV thrombolysis to patients with minor stroke and elderly patients over 80 years old. Debates about the optimal dose of IV tPA according to the ethnic population is still ongoing. Further evidences for the efficacy of intra-arterial thrombolysis have also accumulated, including the application of various novel mechanical devices with promising results. Thus update of guideline became necessary and we revise the acute stroke management guideline, focusing on the thrombolytic therapy.
Aged ; Humans ; Stroke ; Thrombolytic Therapy ; Tissue Plasminogen Activator

PURPOSE: The purpose of this study was to evaluate clinical and radiological results after osteosynthesis of distal phalangeal nonunion with bone defect by using matchstick-shaped bone graft. MATERIALS AND METHODS: Fifteen distal phalangeal nonunions were treated with osteosynthesis using matchstick-shaped bone graft from September, 2009 to October, 2010. The mean age of patients was 36.1 years (range: 15 to 56 years). This study included 4 women and 10 men. The mean duration of follow-up was 16.8 months (range: 12 to 20 months). We evaluated radiographs, and measured postoperative visual analogue scale (VAS) score and pinch power. RESULTS: Union was achieved in all cases at a mean of postoperative 5.5 weeks (range: 4 to 6 weeks). The mean VAS score improved from 7.25 (range: 5 to 9) to 1.0 (range: 0 to 2) postoperatively. The mean postoperative pinch power was 7.75 lbs (range: 4 to 13 lbs) compared to opposite pinch power of 9.63 lbs (range: 5 to 15 lbs). There was no scar problem, pinch pain and other complication. CONCLUSION: Steosynthesis of distal phalageal nonunion with bone defect by using matchstick-shaped bone graft is technically feasible and simple in that it does not need volar incision and debridement. It is considered as an effective treatment option in distal phalangeal nonunion with bone defect.
Bone Transplantation ; Cicatrix ; Debridement ; Female ; Follow-Up Studies ; Humans ; Male ; Transplants

BACKGROUND/AIMS: Chronic hepatitis B infection is a common cause of secondary membranous nephropathy (MN) in endemic areas. Lamivudine treatment improves renal outcome in patients with hepatitis B virus-associated MN (HBV-MN), but prolonged use leads to the emergence of lamivudine-resistant variants. We describe our experience treating lamivudine-resistant and other strains of HBV-MN with new antiviral drugs. METHODS: Of the 89 patients biopsied and diagnosed with MN from 1996 to 2011, 10 positive for hepatitis B surface antigen were recruited for this study. We investigated the clinical courses, therapeutic responses, and prognoses of patients with HBV-MN. RESULTS: The incidence of HBV-MN among the original 89 patients was 11.2%. Of these patients, four were treated with supportive care and six with antiviral drugs. One of the four patients treated with supportive care had a spontaneous remission. Four of the six patients treated with antiviral drugs were given lamivudine, and the other two were given entecavir. Two of the four patients treated with lamivudine achieved complete remission with seroconversion (i.e., development of anti-hepatitis B e antigen antibodies), whereas the other two had lamivudine-resistant strains, which were detected at 22 and 23 months after lamivudine treatment, respectively. We added adefovir to the treatment regimen for one of these patients, and for the other patient we substituted clevudine for lamivudine. Both of these patients experienced complete remission, as did the two patients initially treated with entecavir, neither of whom showed resistance to the drug. CONCLUSIONS: New nucleoside analogues, such as entecavir, adefovir, and clevudine, can be effective for treatment of HBV-MN, including lamivudine-resistant strains.
Adenine/analogs & derivatives/therapeutic use ; Adult ; Antiviral Agents/*therapeutic use ; Arabinofuranosyluracil/analogs & derivatives/therapeutic use ; Drug Resistance, Viral ; Female ; Glomerulonephritis, Membranous/*drug therapy/*etiology ; Guanine/analogs & derivatives/therapeutic use ; Hepatitis B, Chronic/*complications/*drug therapy ; Humans ; Lamivudine/*therapeutic use ; Male ; Middle Aged ; Organophosphonates/therapeutic use ; Young Adult

Since the publication of the first edition of Korean clinical practice guidelines of secondary stroke prevention, encouraging data from recent large clinical trials and meta-analysis have led us to consider many therapeutic options in the treatment of symptomatic extracranial carotid stenosis. Accordingly, the writing group of Clinical Research Center for Stroke decided to provide recent views on the therapeutic revascularization of extracranial carotid stenosis, and timely evidence-based recommendations. In this updated version, new evidences about carotid angioplasty/stenting, treatment timing, and perioperative preparation are given, and qualifying conditions for operator are elucidated. This refinement was based on current consensus between Korean Society of Intravascular Neurosurgery, Korean Society of Interventional Neuroradiology, and Korean Society of Cerebrovascular Surgery and approved by Korean Stroke Society, Korean Neurological Association, and Korean Society of Geriatric Neurology. These recommendations are subject to future correction based on new evidences from ongoing and future studies.
Carotid Arteries ; Carotid Stenosis ; Consensus ; Endarterectomy, Carotid ; Neurology ; Neurosurgery ; Publications ; Secondary Prevention ; Stroke ; Writing