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Immunosorbent assay is the gold standard diagnostic technique for the detection of protein biomarkers. However, this technique tends to have low sensitivity and requires laborious manipulation. Although advanced CRISPR-Cas-based biosensors offer advantages of simplicity, low cost and high accuracy, the synergy of using CRISPR-Cas-assisted dual signal amplification system for rapid diagnosis of protein biomarkers remains scarce. In this work, we report a synergetic signal amplification system comprising CRISPR-Cas12a and nicking enzyme-free strand displacement amplification (SDA) technique for accurate detection of prostate-specific antigen (PSA). The presence of PSA will initiate the nicking enzyme-free SDA process, generating amplicons that can be recognized by the CRISPR-Cas12a system. The activated CRISPR-Cas system will then mediate trans-ssDNA cleavage of neighboring linker DNA, which unlocks the gold nanoparticles (AuNPs) signal probes and gives a distance-dependent colorimetric readout. This assay could detect PSA in aqueous buffer sensitively and selectively with a limit of detection (LOD) down to 0.030 ng mL -1 . Importantly, this assay was successfully applied for discriminating four blood samples from prostate cancer patients among thirteen blood samples from normal individuals/cancer patients accurately. This work will open an avenue for the development of SDA-CRISPR-AuNPs hybrid sensing systems, offering great potential for the development of non-invasive point-of-care diagnostic tools for prostate cancer., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Introduction: The aim of this study was to investigate the diagnostic value of percutaneous computed tomography (CT)-guided coaxial transthoracic needle biopsy (TNB) for small pulmonary lesions (≤ 3 cm) with persistent ground-glass opacity (GGO)., Methods: From January 2004 to February 2010, consecutive patients with persistent small GGO lesions (≤ 3 cm) who underwent CT-guided TNB were analyzed. The pathologic results of CT-guided TNB were compared with final diagnoses, and the GGO percentage on CT was correlated with the stromal invasion in surgical pathology., Results: We performed CT-guided TNB on 1612 patients during the study period. Among them, 55 patients had persistent small GGO lesions (size range 0.5-3.0 cm; 1.72 ± 0.73 cm), 47 were diagnosed with lung adenocarcinomas, and 8 with benign nonspecific lesions. Minor procedure-related complications occurred in 26 patients (47.3%) with small pneumothorax and 11 patients (20.0%) with self-limited mild hemoptysis. The final diagnoses of the 43 patients receiving lobectomy were invasive adenocarcinoma (n = 23), pure bronchioloalveolar carcinoma (n = 19), and mucosa-associated lymphoid tissue lymphoma (n = 1). The diagnostic accuracy was 93.0% (40/43) using CT-guided TNB. Pure GGO lesions had a higher incidence of pure bronchioloalveolar carcinoma than GGO-dominant lesions (70.0% versus 21.7%; p = 0.004). Compared with surgical pathology, stromal invasion was underestimated in 43.5% (10/23) of the TNB specimens, especially in pure GGO lesions (83%, 5/6). In logistic regression analysis, the GGO percentage correlated inversely with stromal tumor invasion (p = 0.0028)., Conclusions: CT-guided coaxial TNB is a safe and useful method for diagnosing small (≤ 3 cm) persistent GGO lesions. Stromal invasion may be underestimated by TNB in GGO lesions.

Generation of a floxed Bmp2/4 osteoblast cell line is a valuable tool for studying the modulatory effects of Bmp2 and Bmp4 on osteoblast differentiation as well as relevant molecular events. In this study, primary floxed Bmp2/4 mouse osteoblasts were cultured and transfected with simian virus 40 large T-antigen. Transfection was verified by polymerase chain reaction (PCR) and immunohistochemistry. To examine the characteristics of the transfected cells, morphology, proliferation and mineralization were analyzed, expression of cell-specific genes including Runx2, ATF4, Dlx3, Osx, dentin matrix protein 1, bone sialoprotein, osteopontin, osteocalcin, osteonectin and collagen type I was detected. These results show that transfected floxed Bmp2/4 osteoblasts bypassed senescence with a higher proliferation rate, but retain the genotypic and phenotypic characteristics similar to the primary cells. Thus, we for the first time demonstrate the establishment of an immortalized mouse floxed Bmp2/4 osteoblast cell line.

Objectives: The aims of this study were to describe the trends of ventricular fibrillation (VF) out-of-hospital cardiac arrest in Rochester, Minnesota, since 1985 and to determine coexistent trends in implantable cardioverter defibrillator (ICD) placement and termination of potentially lethal ventricular arrhythmias that might explain, at least in part, a declining incidence trend., Background: The incidence of VF out-of-hospital cardiac arrest treated by emergency medical services (EMS) personnel has declined over the past decade. Because VF out-of-hospital cardiac arrest occurs primarily in the setting of severe coronary artery disease, primary and secondary prevention strategies may account in part for the decline. In particular, ICD use in large primary and secondary prevention clinical trials in patients at high risk of sudden death has demonstrated that these devices improve survival., Methods: All residents of the City of Rochester, Minnesota, who presented with a VF out-of-hospital cardiac arrest from 1985 to 2002, identified and treated by EMS, were included in the study. In addition, residents of the City of Rochester who received their first ICD implant from 1989 to 2002 were identified. From the ICD records, general demographics, etiology of heart disease, comorbid medical disease, and indication for ICD placement were abstracted. Follow-up data obtained from this population included ICD shocks, the underlying rhythm disturbance, and death., Results: The overall incidence of EMS-treated VF out-of-hospital cardiac arrest in Rochester during the study period was 17.1 per 100,000 [95% confidence interval (CI) 15.1-19.4]. The incidence has decreased significantly (P < 0.001) over the study period: 1985-1989: 26.3/100,000 (95% CI 21.0-32.6), 1990-1994: 18.2/100,000 (95% CI 14.1-23.1), 1995-1999: 13.8/100,000 (95% CI 10.4-17.9), 2000-2002: 7.7/100,000 (95% CI 4.7-11.9). One hundred ten patients received an ICD. The placement of ICDs also has increased dramatically over the past 10 years: 1990-1994: 5.0/100,000 to 2000-2002: 20.7/100,000 (P < 0.001). ICDs terminated VF or fast ventricular tachycardia (<270 ms) in 22 patients. Termination of these potentially fatal arrhythmias has shown a trend toward an increase over the study period: 1990-1994: 1.1/100,000 to 2000-2002: 3.5/100,000 (P = 0.06)., Conclusions: The incidence of VF out-of-hospital cardiac arrest is declining. In contrast, the rates of ICD placement and ICD termination of ventricular tachycardia or VF are markedly increasing. Sudden death preventive strategies are multifactorial. These observations suggest that ICD termination of potentially lethal ventricular arrhythmias may contribute to the lower incidence of VF out-of-hospital cardiac arrest.

Although the efficacy of implantable cardioverter defibrillators (ICDs) has been demonstrated in randomized clinical trials, implantation and survival rates have not been reported for a defined population. We performed a retrospective cohort analysis of Olmsted County, Minnesota residents (n = 70) who received their first ICD between 1 January 1989 and 31 December 1999. The ICD implantation rate increased from approximately 2.5/100,000 (95% confidence interval [CI], 0.9-4.1) in the first 4 years to 11.5/100,000 (95% CI, 6.7-16.2) in the last 2 years. Twenty-three patients (33%) received an appropriate ICD shock during the observation period. Based on these data, ICDs are estimated to reduce total mortality rates in this population by 0.3%. We conclude that, in patients drawn from a community setting with AHA/ACC class I indications for ICD implantation, implantation of ICDs appears to be highly efficacious in aborting potentially fatal events.

Number needed to treat (NNT)-the inverse of the absolute risk reduction resulting from an intervention-was introduced as a yardstick to describe the harm as well as the benefit of therapeutic maneuvers. Analysis using NNT works well when comparing two or more interventions that have their impact over the same period of time in similar populations or patients. Under other conditions, however, analysis based on NNT can produce results that diverge widely from the impact that the interventions can be expected to have on risk of death. This can happen either for entire populations or for an individual when comparing NNTs for interventions which have their effects on different subsets of the population or when comparing interventions which have their effects over different periods of time. We demonstrate how this can occur by comparing the NNTs and effect of intervention on deaths in a population for automatic implantable cardioverter defibrillators (AICDs), heart transplantation, and cholesterol lowering through nutritional intervention with plant stanol ester.