New insights into the pathogenesis of intestina dysfunction:secretory diarrhea and cystic fibrosis

INTRODUCTIONA major function of the intestinal epithelium is to controlthe amount of fluid entering into and being absorbed fromthe lumen.In healthy conditions,net fluid movementfollows an absorptive vector,although significant secretionalso takes place to subserve digestive function.Thus。

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.

冬虫夏草的成熟伴随着冬虫夏草子座中多个基因突变型冬虫夏草菌表达的变化

目的:检测冬虫夏草成熟过程中冬虫夏草菌转换突变和颠换突变基因型在子座中表达的动态变化。方法:采用MassARRAY单核苷酸多态性(single nucleotide polymorphism,SNP)质谱基因分型法,根据冬虫夏草菌基因内转录间隔区(internal transcribed spacer,ITS)序列中大量散在的点突变,设计了8个SNP延伸引物,其中5个位于rDNA ITS1和ITS2段:067721-211,067721-240,067721-477,067721-531和067721-581,用于检验GC和AT偏倚基因型,但不区分两个AT偏倚基因型;另外3个延伸引物位于rDNA 5.8S段:067740-324,067740-328和067740-360,用于区分两个AT偏倚基因型。应用基质辅助激光解吸附电离飞行时间质谱法(matrix-assisted laser desorption/ioniza-tion-time of flight,MALDI-TOF)测定各个SNP位点的单核苷酸延伸反应,观察未成熟和成熟冬虫夏草子座中转换突变和颠换突变基因型表达的动态变化。结果:冬虫夏草子座rDNA的8个SNP位点的质谱图显示复杂的动态变化。067721-211位点的AT偏倚型等位基因的分子浓度明显高于GC偏倚型,但随着冬虫夏草的成熟,这个位点的AT型等位基因消失。多个SNP位点的质谱图中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,有些甚至超过GC和AT偏倚基因型等位基因的峰高;而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。在区分两个AT偏倚基因型的研究中,AB067744和AB067740两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大幅度降低,而AB067740基因型等位基因的峰高大幅度升高。结论:在冬虫夏草子座中共存的至少5个冬虫夏草菌突变基因型的表达随着冬虫夏草的成熟呈现动态变化,这些突变基因型菌的存在和它们的动态变化对于冬虫夏草子座的萌发和成熟具有重要意义。

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。

Gastrointestinal tract access for urological natural orifice transluminal endoscopic surgery

We conducted a literature review of natural orifice transluminal endoscopic surgery(NOTES),focusing on urologic procedures with gastrointestinal tract access,to update on the development of this novel surgical approach.As part of the methods,a comprehensive electronic literature search for NOTES was conducted using Pub Med and Cochrane Library from March 2002 to February 2016 for papers reporting urologic procedures performed utilizing gastrointestinal tract access.A total of 11 peer-reviewed studies examining utility of gastrointestinal access for NOTES urologic procedures were noted,with the first report in 2007.The procedures reported in the studies were total/radical nephrectomy,partial nephrectomy,adrenalectomy,and prostatectomy.The transgastric approach was identified in five studies examining total/radical nephrectomy(n = 2),partial nephrectomy(n = 1),partial cystectomy(n = 1),and adrenalectomy(n = 1).Six studies evaluated transrectal approach for NOTES,describing total/radical nephrectomy(n = 3),partial nephrectomy(n = 1),robotic nephrectomy with adrenalectomy(n = 1) and prostatectomy(n = 1).Feasibility was reported in all studies.Most studies were preclinical and acute,and limited by concerns regarding restricted instrumentation and infection risk.We concluded that gastrointestinal access for urologic NOTES demonstrates promise as described by outlined feasibility studies in preclinical models.Nonetheless,clinical application awaits further advancements in surgical technology and concerns regarding infectious potential.

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.

赢率-医学研究中一个直观而易于解释的综合结果（英文）

对于医学研究中的多个结果,研究人员总是需要选择是否使用一个综合性结果(综合多个结果后所得的)作为主要结果。在本文中,我们回顾了治疗效果的一个新测量方式-赢率,它可以很容易地用于具有多个优先级结果的研究中。并且,我们在这块领域也提出了一些研究课题。

Mechanisms for amplified mediator release from colonic mast cells:Implications for intestinal inflammatory diseases

The mast cell is an enigmatic cell type whose physiological function has preoccupied large numbers of investigators for decadest. Some have concluded that the absence of mast cells is incompatible with life, at least in humans, because no human conditions have been documented where these cells are absent from the body. On the other hand, mice harboring specific mutations in certain growth factors, or their receptors, that

Modern advances in reducing anxiety and pain associated with cystoscopy:Systematic review

AIM:To investigate if music reduces anxiety and pain in the Veterans Affairs population undergoing flexible cystoscopy. METHODS:This study was reviewed and approved by the University of California,San Diego Human Research Protections Program Institutional Review Board. Patients were prospectively randomized to undergo flexible cystoscopy with or without music. Thirty-eight patients were randomized into either the No Music group(n = 24) or the Music group(n = 14). We used the state-trait anxiety inventory and the visual analog pain scale,respectively. Statistics were generated and compared using an independent t-test and chi-squared tests. P values < 0.05 were considered statistically significant. Outpatient cystoscopy is a safe and useful procedure employed frequently in Urology for diagnosis and evaluation of genitourinary pathologies. However,cystoscopy-related distress cannot be ignored. Three components of outpatient cystoscopy have been evaluated to improve the cystoscopic experience:local anesthetic control,cystoscopic equipment redesign and environmental modification. We reviewed the literature pertaining to these modifications. RESULTS:The mean age was 65.3 and 67.1 years for men in the No Music and Music groups,respectively.Although,the majority of patients in each group selfidentified as Caucasians(66%),African American,Hispanic and other ethnicities represented 13%,8% and 13% respectively. The majority of patients(68%) reported experiencing hematuria. Thirty-four percent had a history of bladder cancer,and eighteen percent had a history of prostate cancer. Ten patients(26%) admitted to taking antidepressants. Physiologic parameters that correlated to pain and anxiety(systolic blood pressure,diastolic blood pressure,and heart rate) were statistically similar in both groups prior to and after flexible cystoscopy. The median delta anxiety between the No Music and Music groups were not significantly different(0.78 vs-1.46),and the pain scores between the No Music and Music groups(1.5 vs 1.6) were not statistically different(P = 0.28 and P = 0.92,respectively).CONCLUSION:Preliminary results demonstrate that music does not reduce anxiety or pain associated with flexible cystoscopy.

New insights into the pathogenesis and treatment of non-viral hepatocellular carcinoma:a balancing act between immunosuppression and immunosurveillance

Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths worldwide.HCC initiates as a consequence of chronic liver damage and inflammation caused by hepatitis B and C virus infections,excessive alcohol con sumption,or non-alcoholic fatty liver disease(NAFLD).Until recently,no effective treat-ments for advanced HCC were available and the 5-year survival rate had remained below 8%for many years.New insights into the mechanisms that drive the development of NAFLD-related HCC indicate that loss of T-cell-mediated immunosurveillance plays a cardinal role in tumor growth and malignant progression,in addition to previously identifhed inflammation-driven compensatory proliferation.Recently completed groundbreaking clinical studies have shown that treatments that restore antitumor immunity represent a highly effective therapeutic option for approximately 20%of advanced HCC patients.Understanding the causes of inflammation-driven immunosuppression and immune system dysfunction in the 80%of patients who fail to reignite antitumor immunity despite treatment with checkpoint inhibitors should lead to further and even more dramatic improvements in HCC immunotherapy.

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.

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.

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。

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.

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.