Epigenetic N6-methyladenosine modification of RNA and DNA regulates cancer

The biological roles of N6 methylation of nucleic acids have been extensively studied.Adenine methylation of RNA is the most prevalent RNA modification and has widespread effects on RNA splicing,translation,localization,and stability.Aberrant dynamic regulation of RNA N6-methyladenosine(m6 A)has been reported in numerous human diseases,including several cancers.In recent years,eukaryotic DNA N6-methyladenosine(6 mA)has also been reported and implicated in cancer progression and tumorigenesis.In this review,we summarize the contributions of N6-methyladenosine modification to cancer biology and pathogenesis in the context of both RNA and DNA.We also highlight the clinical relevance of targeting these modifications as a therapeutic strategy for cancer.

Long-Term Electrophysiological and Behavioral Analysis on the Improvement of Visual Working Memory Load, Training Gains, and Transfer Benefits

Recent evidence demonstrates that with training, one can enhance visual working memory (VWM) capacity and attention over time in the near transfer tasks. Not only do these studies reveal the characteristics of VWM load and the influences of training, they may also provide insights into developing effective rehabilitation for patients with VWM deficiencies. However, few studies have investigated VWM over extended periods of time and evaluated transfer benefits on non-trained tasks. Here, we combined behavioral and electroencephalographical approaches to investigate VWM load, training gains, and transfer benefits. Our results reveal that VWM capacity is directly correlated to the difference of event-related potential waveforms. In particular, the “magic number 4” can be observed through the contralateral delay amplitude and the average capacity is 3.25-item over 15 participants. Furthermore, our findings indicate that VWM capacity can be improved through training;and after training exercises, participants from the training group are able to dramatically improve their performance. Likewise, the training effects on non-trained tasks can also be observed at the 12th week after training. Therefore, we conclude that participants can benefit from training gains, and augmented VWM capacity sustained over long periods of time on specific variety of tasks.

A Pilot Trial Testing the Feasibility of Using Molecular-Guided Therapy in Patients with Recurrent Neuroblastoma

Background: Neuroblastoma is the most common extracranial solid tumor in children, and treatment options for recurrent neuroblastoma are limited. Using molecular profiling to target the molecular vulnerabilities of neuroblastoma with existing therapeutic agents may result in a rational, data-driven approach with potential to improve clinical outcomes. Methods: The primary objective of this pilot study was to evaluate the feasibility of supporting real-time treatment decisions through predictive modeling of genome-wide mRNA gene expression data from neuroblastoma tumor biopsies. Feasibility was defined as completion of tumor biopsy, histopathological evaluation, RNA extraction and quality control, gene expression profiling within a CLIA-certified laboratory, bioinformatic analysis, generation of a drug predicttion report, molecular tumor board review yielding a formulated treatment plan, and independent medical monitor review within a 2-week period. Results: Five patients with multiply relapsed or refractory neuroblastoma were enrolled between April and June 2010. All biopsies passed histopathology and RNA quality control. Generation of gene expression data and its analysis (3-7 days), reports which linked this data into medically actionable drug candidates (1-5 days), molecular tumor board (1-3 days) and independent medical monitor review (1 day) were all completed in real-time. The average time was 10.5 days for all patients. Conclusion: This study shows that it is feasible to create therapeutic treatment plans based on genomic profiling in less than 12 days. This warrants further testing in a Phase I study to determine safety of predicted treatments and evaluate whether the information obtained in these analyses would result in patient benefit.

The Effect of Retinoic Acid on Neutrophil Innate Immune Interactions With Cutaneous Bacterial Pathogens

Vitamin A and its biologically active derivative,retinoic acid(RA),are important for many immune processes.RA,in particular,is essential for the development of immune cells,including neutrophils,which serve as a front-line defense against infection.Although vitamin A deficiency has been linked to higher susceptibility to infections,the precise role of vitamin A/RA in host-pathogen interactions remains poorly understood.Here,we provided evidence that RA boosts neutrophil killing of methicillin-resistant Staphylococcus aureus(MRSA).RA treatment stimulated primary human neutrophils to produce reactive oxygen species,neutrophil extracellular traps and the antimicrobial peptide cathelicidin(LL-37).Because RA treatment was insufficient to reduce MRSA burden in an in vivo murine model of skin infection,we expanded our analysis to other infectious agents.RA did not affect the growth of a number of common bacterial pathogens,including MRSA,Escherichia coli K1 and Pseudomonas aeruginosa;however,RA directly inhibited the growth of group A Streptococcus(GAS).This antimicrobial effect,likely in combination with RA-mediated neutrophil boosting,resulted in substantial GAS killing in neutrophil killing assays conducted in the presence of RA.Furthermore,in a murine model of GAS skin infection,topical RA treatment showed therapeutic potential by reducing both skin lesion size and bacterial burden.These findings suggest that RA may hold promise as a therapeutic agent against GAS and perhaps other clinically significant human pathogens.

The differences and similarities between two-sample t-test and paired t-test

临床研究中经常遇到比较实验组和对照组之间的结果。双样本t检验(又称为独立样本t检验)和配对t检验可能是运用于比较两个样本之间均值的最广泛的统计方法。然而,这两种方法的运用会产生混淆,从而导致使用不当。本文中,我们讨论了这两种t检验之间的异同性,并运用三个范例来阐述双样本t检验和配对t检验的计算过程。

Upregulation of RIN3 induces endosomal dysfunction in Alzheimer’s disease

Background In Alzheimer’s Disease(AD),about one-third of the risk genes identified by GWAS encode proteins that function predominantly in the endocytic pathways.Among them,the Ras and Rab Interactor 3(RIN3)is a guanine nucleotide exchange factor(GEF)for the Rab5 small GTPase family and has been implicated to be a risk factor for both late onset AD(LOAD)and sporadic early onset AD(sEOAD).However,how RIN3 is linked to AD pathogenesis is currently undefined.Methods Quantitative PCR and immunoblotting were used to measure the RIN3 expression level in mouse brain tissues and cultured basal forebrain cholinergic neuron(BFCNs).Immunostaining was used to define subcellular localization of RIN3 and to visualize endosomal changes in cultured primary BFCNs and PC12 cells.Recombinant flag-tagged RIN3 protein was purified from HEK293T cells and was used to define RIN3-interactomes by mass spectrometry.RIN3-interacting partners were validated by co-immunoprecipitation,immunofluorescence and yeast two hybrid assays.Live imaging of primary neurons was used to examine axonal transport of amyloid precursor protein(APP)andβ-secretase 1(BACE1).Immunoblotting was used to detect protein expression,processing of APP and phosphorylated forms of Tau.Results We have shown that RIN3 mRNA level was significantly increased in the hippocampus and cortex of APP/PS1 mouse brain.Basal forebrain cholinergic neurons(BFCNs)cultured from E18 APP/PS1 mouse embryos also showed increased RIN3 expression accompanied by early endosome enlargement.In addition,via its proline rich domain,RIN3 recruited BIN1(bridging integrator 1)and CD2AP(CD2 associated protein),two other AD risk factors,to early endosomes.Interestingly,overexpression of RIN3 or CD2AP promoted APP cleavage to increase its carboxyl terminal fragments(CTFs)in PC12 cells.Upregulation of RIN3 or the neuronal isoform of BIN1 increased phosphorylated Tau level.Therefore,upregulation of RIN3 expression promoted accumulation of APP CTFs and increased phosphorylated Tau.These effects by RIN3 was rescued by the expression of a dominant negative Rab5(Rab5S34N)construct.Our study has thus pointed to that RIN3 acts through Rab5 to impact endosomal trafficking and signaling.Conclusion RIN3 is significantly upregulated and correlated with endosomal dysfunction in APP/PS1 mouse.Through interacting with BIN1 and CD2AP,increased RIN3 expression alters axonal trafficking and procession of APP.Together with our previous studies,our current work has thus provided important insights into the role of RIN3 in regulating endosomal signaling and trafficking.

Extrinsic inhibitors in axon sprouting and functional recovery after spinal cord injury

The limited axonal growth after central nervous system （CNS） injury such as spinal cord injury presents a major challenge in promoting repair and recovery. The literature in axonal repair has focused mostly on frank regeneration of injured axons. Here, we argue that sprouting of uninjured axons, an innate repair mech- anism of the CNS, might be more amenable to modulation in order to promote functional repair. Extrinsic inhibitors of axonal growth modulate axon sprouting after injury and may serve as the first group of therapeutic targets to promote functional repair.

Genetic Characterization of Streptococcus pyogenes emm89 Strains Isolated in Japan From 2011 to 2019

Invasive infection caused by Streptococcus pyogenes emm89 strains has been increasing in several countries linked to a recently emergent clade of emm89 strains,designated clade 3.In Japan,the features of emm89 S.pyogenes strains,such as clade classification,remains unknown.In this study,we collected emm89 strains isolated from both streptococcal toxic shock syndrome(STSS)(89 STSS isolates)and noninvasive infections(72 non-STSS isolates)in Japan from 2011 to 2019,and conducted wholegenome sequencing and comparative analysis,which resulted in classification of a large majority into clade 3 regardless of disease severity.In addition,invasive disease-associated factors were found among emm89 strains,including mutations of control of virulence sensor,and absence of the hylP1 gene encoding hyaluronidase.These findings provide new insights into genetic features of emm89 strains.

Implementation and evaluation of Exercise is Medicine in primary care clinics within a large academic health system

Objective Exercise is Medicine(EIM)is a global initiative encouraging healthcare providers to routinely assess and promote physical activity(PA)among patients.The objective of this study was to evaluate the feasibility,adoption,implementation and effectiveness of EIM from patient,clinician and healthcare staff perspectives using a combination of electronic health record(EHR),survey and interview data.Design This study used a combination of the Practical Robust Implementation and Sustainability Model(PRISM)and the Learning Evaluation model to implement EIM.Data captured from the EHR,including Physical Activity Vital Sign(PAVS)scores,and data collected from qualitative surveys and interviews were used to evaluate the programme’s Reach,Effectiveness,Adoption,Implementation,Maintenance(RE-AIM,which is embedded within PRISM)from provider,staff and patient perspectives.Setting Five primary care clinics within a large academic health system.Participants A total of 24443 patients from all participating clinics had at least one PAVS score during the study period.A total of 17 clinicians completed surveys,and 4 clinicians,8 medical assistants and 9 patients completed interviews.Results Implementation fidelity metrics varied widely between components and across clinics but were generally consistent over time,indicating a high degree of programme maintenance.Fidelity was highest during the first 6 months of the COVID-19 pandemic when most visits were virtual.Mean PAVS scores increased from 57.7(95%CI:56 to 59.4)to 95.2(95%CI:91.6 to 98.8)min per week at 6 months for patients not meeting PA guidelines at baseline and decreased from 253.84(95%CI:252 to 255.7)to 208.3(95%CI:204.2 to 212.4)min per week at 6 months for patients meeting PA guidelines at baseline.After EIM implementation,clinician-estimated time spent discussing PA with patients increased for 35%of providers and stayed the same for 53%.Conclusion Overall,this study established EIM’s feasibility,adoption,implementation and maintenance in routine primary care practice within a large academic health system.From a population health perspective,EIM is a model to emulate to help primary care providers efficiently address healthy lifestyle behaviours in routine primary care visits.

至少6个突变基因型冬虫夏草菌在冬虫夏草子座中表达的动态变化

本研究检测了在冬虫夏草成熟过程中突变基因型冬虫夏草菌在子座中表达的动态变化。根据冬虫夏草菌基因内转录间隔区(ITS)序列中大量散在的点突变,设计了8个单核苷酸多态性(SNP)延伸引物,采用SNP质谱基因分型法测定各个SNP位点的单核苷酸延伸反应,观察冬虫夏草子座中转换突变和颠换突变基因型菌在冬虫夏草成熟过程中表达的动态变化。其中5个SNP延伸引物(067721-211,067721-240,067721-477,067721-531和067721-581)位于rDNA ITS1和ITS2段,用于区分GC和AT偏倚基因型,但不区分2个AT偏倚基因型。另外3个延伸引物(067740-324,067740-328和067740-360)位于rDNA 5.8S段,用于区分2个AT基因型。采用PCR+EcoRⅠ限制性酶切法检验2个GC偏倚基因型冬虫夏草菌在冬虫夏草成熟过程中表达的动态变化。结果表明:冬虫夏草子座rDNA ITS1-5.8S-ITS2片段的8个SNP位点的质谱图谱显示冬虫夏草子座中存在至少5个冬虫夏草菌转换突变和颠换突变等位基因。它们的表达在冬虫夏草成熟过程中呈现动态变化。067721-211和067721-477位点的AT偏倚型等位基因的峰高明显高于GC偏倚型;随着冬虫夏草的成熟,这2个位点的AT型等位基因峰高大幅度下降。SNP质谱图不仅显示转换突变的等位基因,颠换突变等位基因也有很高的检出率,它们的峰高在一些位点甚至超过GC和AT基因型等位基因;随着冬虫夏草的成熟,颠换突变等位基因的检出率和峰高趋于下降。在区分2个AT偏倚基因型的研究中,AB067744和AB067740 2个基因型的等位基因同时存在于未成熟冬虫夏草子座中。随着冬虫夏草的成熟,AB067744基因型等位基因的峰高明显降低,而AB067740基因型等位基因的峰高明显升高。PCR产物EcoRⅠ酶切试验发现子座中存在2组GC偏倚型菌,具有完全不同的成熟模式。综上,冬虫夏草子座中存在至少6个突变基因型冬虫夏草菌,其表达随着冬虫夏草的成熟呈现动态变化。这些突变基因型菌表达的动态变化对于冬虫夏草子座的萌发和成熟具有重要菌物学意义。

Direct anti-HCV agents

Unlike human immunodeficiency virus(HIV)and hepatitis B virus(HBV),hepatitis C virus(HCV)infection is a curable disease.Current direct antiviral agent(DAA)targets are focused on HCV NS3/4A protein(protease),NS5 B protein(polymerase)and NS5 A protein.The first generation of DAAs includes boceprevir and telaprevir,which are protease inhibitors and were approved for clinical use in2011.The cure rate for genotype 1 patients increased from 45% to 70% when boceprevir or telaprevir was added to standard PEG-IFN/ribavirin.More effective and less toxic second generation DAAs supplanted these drugs by 2013.The second generation of DAAs includes sofosbuvir(Sovaldi),simeprevir(Olysio),and fixed combination medicines Harvoni and Viekira Pak.These drugs increase cure rates to over 90%without the need for interferon and effectively treat all HCV genotypes.With these drugs the "cure HCV"goal has become a reality.Concerns remain about drug resistance mutations and the high cost of these drugs.The investigation of new HCV drugs is progressing rapidly;fixed dose combination medicines in phase III clinical trials include Viekirax,asunaprevirtdaclatasvirtbeclabuvir,grazoprevirtelbasvir and others.

Acupuncture's Role in Solving the Opioid Epidemic: Evidence, Cost-Effectiveness, and Care Availability for Acupuncture as a Primary, Non-Pharmacologic Method for Pain Relief and Management-White Paper 2017

The United States （U.S.） is facing a national opioid epidemic, and medical systems are in need of non- pharmacologic strategies that can be employed to decrease the public＇s opioid dependence. Acupuncture has emerged as a powerful, evidence-based, safe, cost-effective, and available treatment modality suitable to meeting this need. Acupuncture has been shown to be effective for the management of numerous types of pain conditions, and mechanisms of action for acupuncture have been described and are understandable from biomedical, physiologic perspectives. Further, acupuncture＇s cost-effectiveness can dramatically decrease health care expenditures, both from the standpoint of treating acute pain and through avoiding addiction to opioids that requires costly care, destroys quality of life, and can lead to fatal overdose. Numerous federal regulatory agencies have advised or mandated that healthcare systems and providers offer non-pharmacologic treatment options for pain. Acupuncture stands out as the most evidence-based, immediately available choice to fulfil these calls. Acupuncture can safely, easily, and cost-effectively be incorporated into hospital settings as diverse as the emergency department, labor and delivery suites, andneonatal intensive care units to treat a variety of commonly seen pain conditions. Acupuncture is already being successfully and meaningfully utilized by the Veterans Administration and various branches of the U.S. Military, in some studies demonstrably decreasing the volume of opioids prescribed when included in care.

Current advances in using neurotrophic factors to treat neurodegenerative disorders

Neurotrophic factors are best known for their roles in both development and continued maintenance of the nervous system.Their strong potential to elicit pro-survival and pro-functional responses in neurons of the peripheral and central nervous system make them good drug candidates for treatment of a multitude of neurodegenerative disorders.However,significant obstacles remain and need to be overcome before translating the potential of neurotrophins into the therapeutic arena.This article addresses current efforts and advances in resolving these challenges and provides an overview of roadmaps for future translational research and neurotrophin-based drug developments.

Alcoholic liver disease: A current molecular and clinical perspective

Heavy alcohol use is the cause of alcoholic liver disease(ALD).The ALD spectrum ranges from alcoholic steatosis to steatohepatitis,fibrosis,and cirrhosis.In Western countries,approximately 50%of cirrhosis-related deaths are due to alcohol use.While alcoholic cirrhosis is no longer considered a completely irreversible condition,no effective anti-fibrotic therapies are currently available.Another significant clinical aspect of ALD is alcoholic hepatitis(AH).AH is an acute inflammatory condition that is often comorbid with cirrhosis,and severe AH has a high mortality rate.Therapeutic options for ALD are limited.The established treatment for AH is corticosteroids,which improve short-term survival but do not affect long-term survival.Liver transplantation is a curative treatment option for alcoholic cirrhosis and AH,but patients must abstain from alcohol use for 6 months to qualify.Additional effective therapies are needed.The molecular mechanisms underlying ALD are complex and have not been fully elucidated.Various molecules,signaling pathways,and crosstalk between multiple hepatic and extrahepatic cells contribute to ALD progression.This review highlights established and emerging concepts in ALD clin-icopathology,their underlying molecular mechanisms,and current and future ALD treatment options.

绝经后激素替代治疗 应以最低有效剂量和尽可能短的疗程治疗症状

2007年3月，北美绝经协会（North America Menopause Society，NAMS）公布了绝经后妇女使用激素替代治疗最新状况的说明。NAMS推荐将激素替代治疗作为治疗中重度潮热和阴道萎缩症状的一线用药，十分有效。对于预防骨质疏松性骨折同样有效。但是NAMS建议，激素替代治疗用于这一目的时应充分权衡其潜在危害，也应考虑其他一些已被证明的预防性治疗药物，如二膦酸盐。这些建议清楚、简明，以大量随机对照试验获得的确凿证据为基础。