Real-Time PCR-Based Analysis of the Human Bile MicroRNAome Identifies miR-9 as a Potential Diagnostic Biomarker for Biliary Tract Cancer

Biliary tract cancer (BTC) is often difficult to diagnose definitively, even through histological examination. MicroRNAs (miRNAs) regulate a variety of physiological processes. In recent years, it has been suggested that profiles for circulating miRNAs, as well as those for tissue miRNAs, have the potential to be used as diagnostic biomarkers for cancer. The aim of this study was to confirm the existence of miRNAs in human bile and to assess their potential as clinical biomarkers for BTC. We sampled bile from patients who underwent biliary drainage for biliary diseases such as BTC and choledocholithiasis. PCR-based miRNA detection and miRNA cloning were performed to identify bile miRNAs. Using high-throughput real-time PCR-based miRNA microarrays, the expression profiles of 667 miRNAs were compared in patients with malignant disease (n = 9) and age-matched patients with the benign disease choledocholithiasis (n = 9). We subsequently characterized bile miRNAs in terms of stability and localization. Through cloning and using PCR methods, we confirmed that miRNAs exist in bile. Differential analysis of bile miRNAs demonstrated that 10 of the 667 miRNAs were significantly more highly expressed in the malignant group than in the benign group at P<0.0005. Setting the specificity threshold to 100% showed that some miRNAs (miR-9, miR-302c*, miR-199a-3p and miR-222*) had a sensitivity level of 88.9%, and receiver-operating characteristic analysis demonstrated that miR-9 and miR-145* could be useful diagnostic markers for BTC. Moreover, we verified the long-term stability of miRNAs in bile, a characteristic that makes them suitable for diagnostic use in clinical settings. We also confirmed that bile miRNAs are localized to the malignant/benign biliary epithelia. These findings suggest that bile miRNAs could be informative biomarkers for hepatobiliary disease and that some miRNAs, particularly miR-9, may be helpful in the diagnosis and clinical management of BTC

Evaluation of Transcatheter Arterial Chemoembolization for Unresectable Hepatocellular Carcinoma

The long-term results of transcatheter arterial chemoembolization (LP-TAE) for unresectable hepatocellular carcinoma (HCC) were evaluated in comparison with that of transcatheter arterial chemoinfusion (LP-TAI) and systemic chemotherapy. The cumulative survival rate in 29 patients who received LP-TAE at one-year, two-years and three-years were 70.9%, 54.0% and 25.2%, respectively. In contrast, the cumulative survival rate at one-year in patients who received LP-TAI was 20.6% and those who received systemic chemotherapy was 5.6%. The cumulative survival rate for LP-TAE was significantly higher than those for LP-TAI and systemic chemotherapy (p<0.001). The factor that affected the survival rate for LP-TAE was the size of the tumor. Patients with HCC of less than 5cm in diameter lived significantly longer than those with HCC of more than 5cm in diameter (p<0.05)

Expression of Standard CD44 in Advanced Gastric Cancer: Relationship with Metastasis to Lymph Nodes

Standard CD44 (CD44s) is reported to play an important role in determining the malignant potential of various carcinomas. The aim of the present study was to evaluate CD44s expression in T2-T3 gastric cancer (Japanese Classification of Gastric Cancer stages MP, SS, SE) and the relationship between CD44s expression and clinicopathological parameters. CD44s expression was measured using immunohistochemistry in tumors from 98 patients with primary gastric cancer. Cases were categorized into two groups based on CD44s staining; the CD44s positive group had > 10% positively stained tumor cells and the CD44s negative group had < 10%. CD44s positivity was demonstrated in 59.1% (58/98) of tumors. CD44s expression showed no significant relationship with patient age or gender, or tumor location, size or macroscopic/microscopic classification. However, CD44s expression showed a significantly negative relationship with metastasis to lymph nodes (p < 0.0001). Thus, in T2-T3 gastric cancer, loss of CD44s expression suggests that metastasis of the tumor to lymph nodes is likely

Three Cases of Noninvasive Carcinoma ex Pleomorphic Adenoma of the Parotid Gland and a Literature Survey Focusing on their Clinicopathologic Features

Only 30 cases of non-invasive carcinoma ex pleomorphic adenoma have been reported in the English language literature. Here, we report on three cases of non-invasive carcinoma ex pleomorphic adenoma. Only one of the 33 patients showed recurrence or metastasis after surgery most likely as a result of benign pleomorphic adenoma. Pleomorphic adenoma with focal areas showing malignant changes should be carefully assessed by serial sectioning. The prognosis and therapeutic appoach will depend on evidence of capsular invasion. HER-2/neu is a useful marker in the differential diagnosis of pleomorphic adenoma versus noninvasive carcinoma ex pleomorphic adenoma

Sequencing and Bioinformatics-Based Analyses of the microRNA Transcriptome in Hepatitis B–Related Hepatocellular Carcinoma

MicroRNAs (miRNAs) participate in crucial biological processes, and it is now evident that miRNA alterations are involved in the progression of human cancers. Recent studies on miRNA profiling performed with cloning suggest that sequencing is useful for the detection of novel miRNAs, modifications, and precise compositions and that miRNA expression levels calculated by clone count are reproducible. Here we focus on sequencing of miRNA to obtain a comprehensive profile and characterization of these transcriptomes as they relate to human liver. Sequencing using 454 sequencing and conventional cloning from 22 pair of HCC and adjacent normal liver (ANL) and 3 HCC cell lines identified reliable reads of more than 314000 miRNAs from HCC and more than 268000 from ANL for registered human miRNAs. Computational bioinformatics identified 7 novel miRNAs with high conservation, 15 novel opposite miRNAs, and 3 novel antisense miRNAs. Moreover sequencing can detect miRNA modifications including adenosine-to-inosine editing in miR-376 families. Expression profiling using clone count analysis was used to identify miRNAs that are expressed aberrantly in liver cancer including miR-122, miR-21, and miR-34a. Furthermore, sequencing-based miRNA clustering, but not individual miRNA, detects high risk patients who have high potentials for early tumor recurrence after liver surgery (P = 0.006), and which is the only significant variable among pathological and clinical and variables (P = 0,022). We believe that the combination of sequencing and bioinformatics will accelerate the discovery of novel miRNAs and biomarkers involved in human liver cancer

COVID-19 vaccine effectiveness against severe COVID-19 requiring oxygen therapy, invasive mechanical ventilation, and death in Japan: A multicenter case-control study (MOTIVATE study).

INTRODUCTION: Since the SARS-CoV-2 Omicron variant became dominant, assessing COVID-19 vaccine effectiveness (VE) against severe disease using hospitalization as an outcome became more challenging due to incidental infections via admission screening and variable admission criteria, resulting in a wide range of estimates. To address this, the World Health Organization (WHO) guidance recommends the use of outcomes that are more specific to severe pneumonia such as oxygen use and mechanical ventilation. METHODS: A case-control study was conducted in 24 hospitals in Japan for the Delta-dominant period (August-November 2021; "Delta") and early Omicron (BA.1/BA.2)-dominant period (January-June 2022; "Omicron"). Detailed chart review/interviews were conducted in January-May 2023. VE was measured using various outcomes including disease requiring oxygen therapy, disease requiring invasive mechanical ventilation (IMV), death, outcome restricting to "true" severe COVID-19 (where oxygen requirement is due to COVID-19 rather than another condition(s)), and progression from oxygen use to IMV or death among COVID-19 patients. RESULTS: The analysis included 2125 individuals with respiratory failure (1608 cases [75.7%]; 99.2% of vaccinees received mRNA vaccines). During Delta, 2 doses provided high protection for up to 6 months (oxygen requirement: 95.2% [95% CI:88.7-98.0%] [restricted to "true" severe COVID-19: 95.5% {89.3-98.1%}]; IMV: 99.6% [97.3-99.9%]; fatal: 98.6% [92.3-99.7%]). During Omicron, 3 doses provided high protection for up to 6 months (oxygen requirement: 85.5% [68.8-93.3%] ["true" severe COVID-19: 88.1% {73.6-94.7%}]; IMV: 97.9% [85.9-99.7%]; fatal: 99.6% [95.2-99.97]). There was a trend towards higher VE for more severe and specific outcomes. CONCLUSION: Multiple outcomes pointed towards high protection of 2 doses during Delta and 3 doses during Omicron. These results demonstrate the importance of using severe and specific outcomes to accurately measure VE against severe COVID-19, as recommended in WHO guidance in settings of intense transmission as seen during Omicron