Diagnosis of benign/malignant common bile duct (CBD) stenoses is often difficult. No currently available diagnostic modality ensures absolute accuracy in differentiating malignant from nonmalignant CBD stenoses. Cell-free nucleic acids in bile might serve as potential diagnostic biomarkers. We investigated whether cell-free DNA in bile could distinguish malignant from nonmalignant CBD stenoses. Bile samples were collected from 28 patients undergoing therapeutic endoscopic retrograde cholangiopancreatography at university hospitals for CBD stenosis of malignant (biliary cancer, n=9; pancreatic cancer, n=9) or nonmalignant (n=10) disease. KRAS mutations and DNA methylation were analyzed by quantitative pyrosequencing. Receiver operating characteristic area under the curve (AUC) was used to determine diagnostic accuracy. KRAS mutations were detected in 67% of biliary cancers, 56% of pancreatic cancers, and 10% of the nonmalignant disease. No significant differences were found in DNA methylation levels of CDKN2A, MINT25, SOX17, and miR-34b/c between malignant and nonmalignant diseases. BARHL2 methylation levels were significantly higher in bile samples from patients with malignant CBD stenoses than in those with nonmalignant CBD stenoses. Moreover, bile BARHL2 methylation levels distinguished malignant from nonmalignant CBD stenoses with 88% sensitivity and 100% specificity (AUC=0.95556). Thus, the analysis of bile BARHL2 methylation levels may be useful for the diagnosis of pancreatobiliary malignancies in clinical settings.
Gastric cancer (GC) is one of the most common malignancies and is the leading cause of cancer-related deaths worldwide. CagA, encoded by cytotoxin-associated gene A (CagA), is a major virulence factor of Helicobacter pylori, a gastric pathogen involved in the development of GC. We previously developed a method for DNA methylation analysis, and early detection of GC using gastric washes rather than highly acidic gastric juice. Extracellular vesicles known as exosomes present in the gastric juice may provide an alternative to gastric washes for the molecular detection of GC. We determined tumor-related alterations in the functional molecular contents within the exosomes, which were purified from the gastric juice of patients with GC, and whether the exosomes contained CagA. Using ultracentrifugation, we purified exosomes from the gastric juice of patients with GC and analyzed their expression levels of microRNA-34 (miR-34) by real-time PCR. Methylation levels of the miR-34b/c gene in exosomal DNA were analyzed by bisulfite pyrosequencing. The microvesicles were verified as exosomes by transmission electron microscopy and western blot analysis with the CD9 exosomal marker. Expression levels of miR-34 were lower in tumor-derived exosomes than in non-tumor-derived exosomes. Methylation levels of miR-34b/c were higher in tumor-derived exosomes than in non-tumor-derived exosomes. Concordant miR-34b/c methylation levels were observed between the exosomal and tissue nuclear DNA from patients with GC. CagA was detected in exosomes obtained from the gastric juice of CagA-positive H. pylori-infected patients with GC. These findings suggest the use of gastric juice-derived exosomes as a biomarker for GC in clinical settings in the future.
Background In 2008, Nakajima et al. created an organ damage classification system based on CT finding to guide recommended treatments in patients with hepatic and splenic injury, but its practicality has not yet been investigated. This hepatic and splenic injury grading system, which includes extravasation, pseudoaneurysm and arteriovenous fistula, aims to provide a guideline for initial management decisions. However, no studies have reviewed its usefulness, and thus we attempted to assess the efficacy and usefulness of the grading system. Materials and Methods From April 2006 to December 2015, we retrospectively reviewed 96 splenic and 117 hepatic injury patients and assessed the effectiveness and correlation of the scoring system with the patients’ actual clinical injury. We compared the injury grades (from least severe[grade I] to most severe[grade V]) with the degree of invasiveness of the initial treatment management (least to most invasive: conservative<interventional radiology<surgery). Results The CT-based scoring system showed satisfactory correlation between the degree of severity of management and the injury in 90.6% (87/96) of the splenic and 94.9% (111/117) of the hepatic injury patients. There was a direct correlation between the degree of treatment invasiveness and the scoring grade. Conclusion The CT-based scoring system proposed by Nakajima et al. can be a very useful tool in correlating the grade of splenic and hepatic injury with the invasiveness of the initial treatment and in predicting initial treatment management. However, the scoring system has limitations in that the management decision does not solely depend on imaging findings but also various other factors.
Purpose: To improve the surgical curability of prostate cancer, we introduced minimum incision endoscopic radical prostatectomy with intentional wide resection (MRP–WR) at our hospital. We report the surgical outcomes of MRP–WR in comparison with those of MRP, which was used previously. Subjects and methods: The analysis included 21 patients with prostate cancer who underwent MRP–WR between January 2016 and March 2017 and a control group of 23 patients who underwent MRP between April 2013 and May 2015. The surgical outcomes were retrospectively analyzed by examining clinical parameters, including surgical time, hemorrhage volume, number of lymph nodes removed, duration of indwelling urinary catheterization, duration of hospitalization, positive- surgical margin rate, and initial postoperative serum prostate-specific antigen (PSA) level. Results: Compared with the patients in the MRP group, those in the MRP–WR group had a longer mean surgery time (251 vs. 318 min, P<0.0001), lower mean hemorrhage volume (1199 vs. 292 mL, P<0.0001), larger mean number of removed lymph nodes (3.7 vs. 19.1, P<0.0001), longer duration of indwelling urinary catheterization (6.2 vs. 9.9 days, P=0.0042), and a lower positive- surgical- margin rate (48 vs. 19%, P=0.0443). The initial postoperative PSA level was below the detection limit in 67% of the patients in the MRP–WR group, whereas it was below the detection limit in only 4% in the MRP group, indicating that patients in the MRP–WR group had a significantly lower initial postoperative PSA level (P<0.0001). Conclusion: The newly introduced MRP–WR procedure appears to be superior to conventional MRP in terms of the complete resection rate of prostate cancer, improvement in treatment outcomes, and reduction in hemorrhage.
Background: To investigate the effects of platelet-rich plasma (PRP) on hair growth, the authors used molecular biology to investigate the influence of PRP on cultured human dermal papilla cells (DPC). Methods: PRP-treated DPC were cultured for a week. Gene expression levels of FGF-2, VEGF, BMP-2, Wnt5a, and EFNA3 were measured using PCR. Results: PRP significantly increased FGF-2, VEGF, and BMP-2 gene expression levels in DPC after 2 hours compared to those without PRP. After 24 hours, increased expression of Wnt5a was maintained. However, there was no significant difference in VEGF expression. BMP-2 expression was significantly decreased, but that of EFNA3 did not change. There were no changes in the expression levels of these genes in PRP with exosomes removed. Discussion: These results suggested that increased expression of hair growth-related genes in DPC activates hair follicles in the catagen phase of the hair cycle. Sustained expression of Wnt5a, and the short-term strong expression of BMP2 followed by its significant decrease suppress differentiation and cause atrophy of hair papillae. Conclusions: PRP is not involved in hair growth but is involved in hair development.