Shikonin from Chinese herbal medicine induces GSDME-controlled pyroptosis in tumor cells

ObjectiveTo investigate the potential anti-tumor mechanisms of naphthoquinone compound shikonin (SKN) extracted from the root of Chinese herbal medicine plant lithospermum (Lithospermum erythrorhizon Sieb. & Zucc.).MethodsWe first observed that SKN treatment led to swelling and bubbles in HeLa cells that were similar to the phenotype of cell pyroptosis. Subsequently, the HeLa cells experienced a pyroptotic process with SKN, and this was then assessed using lactate dehydrogenase (LDH) release and propidium iodide (PI)/Hoechst double staining experiments. Pyroptosis is defined as gasdermin-mediated programmed necroptosis. To identify the potential pyroptosis machinery, two strategies were utilized that included a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 screening experiment and a pyroptosis reconstitution assay executed by each of the five known gasdermins (GSDMA-E). Moreover, endogenous cleavage was also detected in a panel of tumor cell lines.ResultsCompared with the control, both the LDH release and PI/Hoechst double-staining experiments suggested that SKN induced perforation and enhancement of the permeability of the cell membranes that resulted in pyroptosis in HeLa cells (P = .028 and P = .032, respectively). In addition, the reconstitution assays in human embryonic kidney 293T (HEK-293T) cells and endogenous cleavage assays in HeLa cells indicated that the pyroptosis was controlled by GSDME. In addition, we also found SKN could trigger pyroptosis in a panel of tumor cell lines in which the cellular morphologies were proportional to the GSDME expression levels. Additionally, the cleavage of GSDME was also detected, and this was indicative of a similar GSDME-mediated mechanism.ConclusionOur study not only explained the molecular mechanism of cytotoxicity of SKN to various tumor cells, but also provided additional information for the potential clinical application of natural naphthoquinone compounds against cancer.     

Step-count outcomes of 13 different activity trackers: Results from laboratory and free-living experiments

BackgroundMany activity trackers have been developed, but steps can still be inconsistent from one monitor to another.Research questionWhat are the differences and associations between the steps of 13 selected consumer-based and research-grade wearable devices during 1 standardized day in a metabolic chamber and 15-day free-living trials?MethodsIn total, 19 healthy adults between 21 and 50 years-old participated in this study. Participants were equipped with 12 accelerometer-based active trackers and one pedometer (Yamasa) in order to monitor the number of steps per day. The devices were worn on the waist (ActiGraph, Omron, Actimarker, Lifedorder, Withings, and Yamasa) or non-dominant wrist (Fitbit, Garmin, Misfit, EPSON, and Jawbone), or placed in a pocket (Omron CaloriScan, and TANITA). Participants performed structured activities over a 24 h period in a chamber (Standardized day), and steps were monitored in the same participants in free-living trials for 15 successive days using the same monitors (free-living days).ResultsWhen the 13 monitors were ranked by their steps, waist-worn ActiGraph was located at the center (7th) of the monitors both in the Standardized (12,252 ± 598 steps/day, mean ± SD) and free-living days (9295 ± 4027 steps/day). The correlation between the accelerometer-based devices was very high (r = 0.87–0.99). However, the steps of Yamasa was significantly lower in both trials than ActiGraph. The wrist-worn accelerometers had significantly higher steps than other devices both trials (P < 0.05). The differences between ActiGraph and Actimarker or Lifecorder was less than 100 steps/day in the Standardized day, and the differences between ActiGraph and Active Style Pro was less than 100 steps/day in the free-living days. Regression equation was also performed for inter-device compatibility.SignificanceStep obtained from the wrist-worn, waist-worn, and pocket-type activity trackers were significantly different from each other but still highly correlated in free-living conditions.     

A Comparison of the Cost-Effectiveness of Lifestyle Interventions in Pregnancy

ObjectivesLifestyle interventions during pregnancy improve maternal and infant outcomes. We aimed to compare the cost-effectiveness of 4 antenatal lifestyle intervention types with standard care.MethodsA decision tree model was constructed to compare lifestyle intervention effects from a novel meta-analysis. The target population was women with singleton pregnancies and births at more than 20 weeks’ gestation. Interventions were categorized as diet, diet with physical activity, physical activity, and mixed (lacking structured diet and, or, physical activity components). The outcome of interest was cost per case prevented (gestational diabetes, hypertensive disorders in pregnancy, cesarean birth) expressed as an incremental cost-effectiveness ratio (ICER) from the Australian public healthcare perspective. Scenario analyses were included for all structured interventions combined and by adding neonatal intensive care unit costs. Costs were estimated from published data and consultations with experts and updated to 2019 values. Discounting was not applied owing to the short time horizon.ResultsPhysical activity interventions reduced adverse maternal events by 4.2% in the intervention group compared with standard care and could be cost saving. Diet and diet with physical activity interventions reduced events by 3.5% (ICER = A$4882) and 2.9% (ICER = A$2020), respectively. Mixed interventions did not reduce events and were dominated by standard care. In scenario analysis, all structured interventions combined and all interventions when including neonatal intensive care unit costs (except mixed) may be cost saving. Probabilistic sensitivity analysis showed that for physical activity and all structured interventions combined, the probability of being cost saving was 58% and 41%, respectively.ConclusionsGovernments can expect a good return on investment and cost savings when implementing effective lifestyle interventions population-wide.     

蛋白质精氨酸甲基转移酶1在肿瘤发生发展中作用的研究进展北大核心

蛋白质精氨酸甲基转移酶1(protein arginine methyltransferase 1,PRMT1)是主要的Ⅰ型精氨酸甲基转移酶,催化一甲基化和不对称二甲基化,其底物被甲基化后参与细胞生物学过程。研究表明,PRMT1在多种恶性肿瘤中异常表达,促进肿瘤细胞的增殖、侵袭和迁移,调控人类多种肿瘤发生发展过程,揭示PRMT1可能成为肿瘤治疗中潜在的生物标志物或靶点。本文对PRMT1的结构、底物、生物学功能、及其在肿瘤发生发展中的作用进行总结,旨在为今后研究PRMT1相关肿瘤提供参考。     

American Cancer Society nutrition and physical activity guideline for cancer survivors

The overall 5-year relative survival rate for all cancers combined is now 68%, and there are over 16.9 million survivors in the United States. Evidence from laboratory and observational studies suggests that factors such as diet, physical activity, and obesity may affect risk for recurrence and overall survival after a cancer diagnosis. The purpose of this American Cancer Society guideline is to provide evidence-based, cancer-specific recommendations for anthropometric parameters, physical activity, diet, and alcohol intake for reducing recurrence and cancer-specific and overall mortality. The audiences for this guideline are health care providers caring for cancer survivors as well as cancer survivors and their families. The guideline is intended to serve as a resource for informing American Cancer Society programs, health policy, and the media. Sources of evidence that form the basis of this guideline are systematic literature reviews, meta-analyses, pooled analyses of cohort studies, and large randomized clinical trials published since 2012. Recommendations for nutrition and physical activity during cancer treatment, informed by current practice, large cancer care organizations, and reviews of other expert bodies, are also presented. To provide additional context for the guidelines, the authors also include information on the relationship between health-related behaviors and comorbidities, long-term sequelae and patient-reported outcomes, and health disparities, with attention to enabling survivors' ability to adhere to recommendations. Approaches to meet survivors' needs are addressed as well as clinical care coordination and resources for nutrition and physical activity counseling after a cancer diagnosis.     

Effect of chlorhexidine digluconate on antimicrobial activity,cell viability and physicochemical properties of three endodontic sealers

ObjectiveAssess the biological and physicochemical properties of AH Plus, BioRoot RCS and Pulp Canal Sealer (PCS) leachates with and without chlorhexidine (CHX).MethodsThe sealers were studied in no contact and 1-minute contact with CHX. For biological properties (antibacterial activity and cytotoxicity), leachates were formed in saline of freshly mixed, 1-, 7- and 28 days set sealers. The antibacterial properties of sealer leachates were investigated for planktonic and biofilm growth of E. faecalis, S. mutans, S.epidermidis and S.aureus. The 3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate murine fibroblast cell viability after exposure to the leachates. The physical properties (water uptake, sorption, solubility, porosity, surface characteristics) of sealers and the pH of the immersion liquid (saline or distilled water) were also assessed over a 28-days period.ResultsCHX improved the antibacterial properties of the sealer leachates and reduced cell viability for all sealer leachates, except for freshly mixed PCS. BioRoot RCS leachates presented the highest antibacterial properties and cell viability with and without CHX contact. PCS was the material most affected by CHX in terms of physical properties, whereas for AH Plus, solubility was increased. CHX did not affect the physical properties of BioRoot RCS, except for solubility that was decreased. CHX contact did not change sealers’ alkalinity in distilled water whereas it increased it for AH Plus and BioRoot RCS in saline.SignificanceCHX improved the antibacterial efficacy of sealer leachates and either compromised or did not affect cell viability. CHX affected to various extent sealers’ physicochemical properties.     

氢氧化钙糊剂与根管内感染控制

感染控制对根管治疗的成功至关重要。尽管随着器械及设备的进步，对根管的清理及成形取得了很大的进展；然而，由于根管系统解剖结构错综复杂，使得获取清洁根管面临各种问题。例如：机械预备的宽度有限，不能彻底清理感染牙本质；冲洗技术的运用虽然可以去除根管器械预备后形成的玷污层，但对细菌内毒素的清除还是难以达到理想的效果，而残留的微生物、毒素等若得不到清除可能会在根尖周组织形成持续感染。因此，除外机械预备和冲洗，根管封药也是感染根管治疗过程中的重要步骤。氢氧化钙糊剂以其高效的抗菌作用在临床得到广泛使用。文章将对根管封药的必要性、氢氧化钙的作用机制及临床应用做一介绍。     

TRIM59 通过结合 BCLAF1 调控人皮肤黑色素瘤 SK-MEL-2 细胞的恶 性生物学行为

目的：探究三结构域蛋白59（TRIM59）调控人皮肤黑色素瘤细胞SK-MEL-2增殖、细胞周期、凋亡及迁移侵袭的作用机制，及其与Bcl2相关转录因子1（BCLAF1)之间的关系。方法：qPCR和WB法检测人表皮黑色素细胞HEMn-LP、人皮肤黑色素瘤细胞SK-MEL-2、UACC903、A375及36例邢台市人民医院2019年2月至2021年7月收集的皮肤黑色素瘤组织中TRIM59的mRNA和蛋白表达，使用脂质体将si-con、si-TRIM59转染至SK-MEL-2细胞中，WB法检测干扰TRIM59表达对细胞中周期蛋白D1（CCND1）、细胞周期素依赖性激酶2（CDK2）、肿瘤抑制蛋白基因（TP53）和 BCLAF1 蛋白表达的影响，CCK-8法、流式细胞术、划痕愈合实验、Transwell实验检测对细胞的活性、凋亡、迁移和侵袭的影响，免疫共沉淀（Co-IP）实验检测对细胞中TRIM59蛋白与BCLAF1结合能力的影响。结果：与HEMn-LP细胞相比，SK-MEL-2、UACC903、A375细胞中TRIM59 mRNA和TRIM59、BCLAF1蛋白均呈高表达（均P<0.05），SK-MEL-2细胞中TRIM59表达水平最高。相较于si-con组和Normal组，沉默TRIM59后，SK-MEL-2细胞的活性显著降低，细胞周期阻滞于G2期，CCND1、CDK2的蛋白表达显著降低，TP53蛋白和细胞凋亡率均显著升高，划痕抑制率明显升高，迁移侵袭细胞数明显降低（均P<0.05）。免疫共沉淀实验结果显示，TRIM59与BCLAF1之间存在蛋白结合关系。TRIM59与 BCLAF1 在肿瘤组织中的表达呈显著的正相关（r=0.878，P<0.001）。结论：干扰TRIM59表达能够抑制人皮肤黑色素瘤SK-MEL-2细胞的增殖、迁移和侵袭而促进凋亡，抑制SK-MEL-2细胞的恶性生物学行为，其机制可能与TRIM59结合BCLAF1有关。