Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.

Neuroprotective effects of Shaoyao Gancao decoction against excitatory damage in PC12 cells based on the Src-NR2-nNOS pathway

Objective To explore the neuroprotective effects of the Shaoyao Gancao decoction(SGD)against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulatingγ-aminobutyric acid(GABA)-glutamate(Glu)homeostasis.Methods N-Methyl-d-aspartic acid(NMDA)was used to establish a PC12 cell excitability injury model.To investigate the neuroprotective effect of SGD,a cell counting kit-8(CCK-8)assay was used to determine PC12 cell viability,Annexin V/Propidium Iodide(Annexin V/PI)double staining was used to determine PC12 cell apoptosis,and Ca^(2+)concentration was observed using laser confocal microscopy.GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions betweenγ-aminobutyric acid(GABA)and NMDA receptors.Additionally,molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway.We analyzed the correlations between the regulatory sites of GABA and NMDA interactions,excitatory neurotoxicity,and brain damage at the molecular level.Results NMDA excitotoxic injury manifested as a significant decrease in cell activity,increased apoptosis and caspase-3 protein expression,and a significant increase in intracellular Ca^(2+)concentration.Administration of SGD,a GABAA receptor agonist(muscimol),or a GABAB receptor agonist(baclofen)decreased intracellular Ca^(2+)concentrations,attenuated apoptosis,and reversed NMDA-induced upregulation of caspase-3,Src,NMDAR2A,NMDAR2B,and nNOS.Unexpectedly,a GABA_(A)receptor antagonist(bicuculline)and a GABA_(B)receptor antagonist(saclofen)failed to significantly increase excitatory neurotoxicity.Conclusions Taken together,these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases,but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.

Health 3.0—The patient-clinician “arabic spring” in healthcare

A growing number of citizen-patients and clinicians use Communication and Self-Managed Health Technologies (CSMHT) in their relationship. Doing so, they shift from the current paradigm of dependency to a co-responsibility paradigm in healthcare. Facing the runaway utilization of health services, we need to think “outside the box” to unblock the system. A Health 3.0 development model of governance that position patients as primary members of the clinicians’ team is presented to map this institutional transformation. At the practical level, an MD 3.0 relational model and a Citizen-Patient 3.0 behavioral profile are presented.

小鼠Reg3α基因cDNA成功转染MIN6细胞株并促其生长

目的建立一过表达Reg3α cDNA的胰岛MIN6细胞株，研究其在低葡萄糖环境中的生长特性。方法将全长Reg3α cDNA插入到质粒载体pcDNA3．1中，并将Reg3α-pcDNA3．1和pcDNA3．1空载体用脂质体法转染胰岛MIN6细胞，分别获得Reg3α cDNA过表达和空载体pcDNA3．1MIN6细胞。应用Real—time PCR和Western blot方法检测这两种细胞在低葡萄糖培养基中Reg3α的表达，并用MTT法测定Reg3α过表达对MIN6细胞生长的影响。结果有三株Reg3α转染的细胞株显示Reg3α的高表达，在空载体转染的MIN6细胞株中未检测到Reg3α的表达，Reg3α mRNA和蛋白水平分别平均升高6～10倍。用Western blot检测发现Reg3α蛋白释放到培养液。采用MTT法测定，与空载体转染的MIN6细胞株相比，三株Reg3α稳定转染细胞株，7d后细胞数目升高2倍。结论本研究结果表明已成功地建立了过表达Reg3α cDNA的胰岛MIN6细胞。Reg3α可能具有内分泌作用，促进胰岛细胞生长。

PTPN22 C1858T与1型糖尿病相关性荟萃分析

利用荟萃分析评价PTPN22基因C1858T多态位点与1型糖尿病(T1DM)发病的相关性。资料来自Medline和中国生物医学文献数据库收录的文献,纳入涉及C1858T多态位点和T1DM相关性的病例对照或家系研究,应用RevMan4·2软件进行统计分析。相关文献未发现显著发表偏倚;在病例对照及家系研究中,T等位基因(P<0·01,OR=1·87和P<0·001,OR=1·66)均增加T1DM发病风险。PTPN22 C1858T位点与T1DM遗传易感性相关;该多态位点仅见于欧洲裔人群,在不同欧洲裔人群中,该位点遗传作用大小(OR)相似。

Systematic review: Preventive and therapeutic applications of metformin in liver disease

Metformin, a biguanide derivative, is the most commonly prescribed medication in the treatment of type 2 diabetes mellitus. More recently, the use of metformin has shown potential as a preventive and therapeutic agent for a broad spectrum of conditions, including liver disease and hepatic malignancies. In this systematic review,we critically analyze the literature behind the potential use of metformin across the spectrum of liver disease and malignancies. The Pub Med and Ovid MEDLINE databases were searched from 2000 to March 2015, using a combination of relevant text words and MeS H terms: metformin and mammalian target of rapamycin, hepatitis B virus(HBV), hepatitis B virus(HCV), nonalcoholic fatty liver disease(NAFLD), hepatocellular carcinoma(HCC) or cholangiocarcinoma. The search results were evaluated for pertinence to the issue of metformin in liver disease as well as for quality of study design. Metformin has a number of biochemical effects that would suggest a benefit in treating chronic liver diseases, particularly in the context of insulin resistance and inflammation. However, the literature thus far does not support any independent therapeutic role in NAFLD or HCV. Nonetheless, there is Level Ⅲ evidence for a chemopreventive role in patients with diabetes and chronic liver disease, with decreased incidence of HCC and cholangiocarcinoma. The use of metformin seems to be safe in patients with cirrhosis, and provides a survival benefit. Once hepatic malignancies are already established, metformin does not offer any therapeutic potential. In conclusion, there is insufficient evidence to recommend use of metformin in the adjunctive treatment of chronic liver diseases, including NAFLD and HCV. However, there is good evidence for a chemopreventive role against HCC among patients with diabetes and chronic liver disease, and metformin should be continued in patients even with cirrhosis to provide this benefit.

Prevalence and predictors of nonalcoholic fatty liver disease in South Asian women with polycystic ovary syndrome

BACKGROUND Polycystic ovary disease(PCOS)may be a risk factor for nonalcoholic fatty liver disease(NAFLD)due to common pathogenetic pathways,including insulin resistance and obesity.Both PCOS and NAFLD are more severe in South Asian women.Data on NAFLD in South Asian women with PCOS are lacking.AIM To investigate prevalence and predictors of NAFLD and liver fibrosis in PCOS patients from South Asia.METHODS We conducted an observational routine screening program by means of transient elastography(TE)with associated controlled attenuation parameter(CAP).NAFLD was defined as CAP≥288 decibels per meter.Significant liver fibrosis(stage 2 and higher out of 4)was defined as TE measurement≥8.0 kilopascals.Elevated alanine aminotransferase(ALT)was defined as ALT>24 IU/L,as per upper limit of normal reported in South Asian women.Biochemical hyperandrogenism was defined as free androgen index>5.Predictors of NAFLD were determined by logistic regression analysis.RESULTS 101 PCOS patients(mean age 36.3 years)with no significant alcohol intake or viral hepatitis were included.Prevalence of NAFLD and significant liver fibrosis was 39.6% and 6.9%,respectively.Elevated ALT was observed in 40%and 11.5%of patients with and without NAFLD,respectively.After adjusting for duration of PCOS and insulin resistance measured by homeostasis model for assessment of insulin resistance,independent predictors of NAFLD were higher body mass index[adjusted odds ratio(aOR)1.30,95% confidence interval(CI):1.13-1.52],hyperandrogenism(aOR:5.32,95%CI:1.56-18.17)and elevated ALT(aOR:3.54,95%CI:1.10-11.47).Lifetime cardiovascular risk was higher in patients with NAFLD compared to those without NAFLD(0.31±0.11 vs 0.26±0.13).CONCLUSION Despite their young age,NAFLD diagnosed by TE with CAP is a frequent comorbidity in South Asian women with PCOS and is strongly associated with higher body mass index and hyperandrogenism.Non-invasive screening strategies could help early diagnosis and initiation of interventions,including counselling on weight loss,cardiovascular risk stratification and linkage to hepatology care where appropriate.

Preoperative selection of patients with colorectal cancerliver metastasis for hepatic resection

Surgical resection of colorectal liver metastases(CRLM) has a well-documented improvement in survival. To benefit from this intervention, proper selection of patients who would be adequate surgical candidates becomes vital. A combination of imaging techniques may be utilized in the detection of the lesions. The criteria for resection are continuously evolving; currently, the requirements that need be met to undergo resection of CRLM are: the anticipation of attaining a negative margin(R0 resection), whilst maintaining an adequate functioning future liver remnant. The timing of hepatectomy in regards to resection of the primary remains controversial; before, after, or simultaneously. This depends mainly on the tumor burden and symptoms from the primary tumor. The role of chemotherapy differs according to the resectability of the liver lesion(s); no evidence of improved survival was shown in patients with resectable disease who received preoperative chemotherapy. Presence of extrahepatic disease in itself is no longer considered a reason to preclude patients from resection of their CRLM, providing limited extra-hepatic disease, although this currently is an area of active investigations. In conclusion, we review the indications, the adequate selection of patients and perioperative factors to be considered for resection of colorectal liver metastasis.

Metformin does not improve survival in patients with hepatocellular carcinoma

AIM: To assess whether metformin, which has a chemopreventive effect in chronic liver disease, has any chemotherapeutic effect in hepatocellular carcinoma.

Disease monitoring strategies in inflammatory bowel diseases: What do we mean by “tight control”?

In recent years,there has been a critical change in treatment paradigms in inflammatory bowel diseases(IBD)triggered by the arrival of new effective treatments aiming to prevent disease progression,bowel damage and disability.The insufficiency of symptomatic disease control and the well-known discordance between symptoms and objective measures of disease activity lead to the need of reviewing conventional treatment algorithms and developing new concepts of optimal therapeutic strategy.The treat-to-target strategies,defined by the selecting therapeutic targets in inflammatory bowel disease consensus recommendation,move away from only symptomatic disease control and support targeting composite therapeutic endpoints(clinical and endoscopical remission)and timely assessment.Emerging data suggest that early therapy using a treat-to-target approach and an algorithmic therapy escalation using regular disease monitoring by clinical and biochemical markers(fecal calprotectin and C-reactive protein)leads to improved outcomes.This review aims to present the emerging strategies and supporting evidence in the current therapeutic paradigm of IBD including the concepts of“early intervention”,“treat-to-target”and“tight control”strategies.We also discuss the real-word experience and applicability of these new strategies and give an overview on the future perspectives and areas in need of further research and potential improvement regarding treatment targets and(“tight”)disease monitoring strategies.

Non-alcoholic fatty liver disease-a chronic disease of the 21^(st) century

Non-alcoholic fatty liver disease（NAFLD） comprises a spectrum of metabolic states ranging from simple steatosis to inflammation with associated fibrosis to cirrhosis. Though accumulation of hepatic fat is not associated with a significant increase in mortality rates, hepatic inflammation is, as this augments the risk of terminal liver disease, i.e.,cirrhosis, hepatic decompensation（liver failure） and/or hepatocellular carcinoma. Disease progression is usually slow, over a decade or more and, for the most part, remains asymptomatic. Recent estimates suggest that the global prevalence of NAFLD is high, about one in four. In most cases, NAFLD overlaps with overweight, obesity,cardiovascular disease and the metabolic syndrome with numerous contributing parameters including a dysregulation of adipose tissue, insulin resistance, type 2 diabetes, changes in the gut microbiome, neuronal and hormonal dysregulation and metabolic stress. NAFLD is diagnosed incidentally, despite its high prevalence. Non-invasive imaging techniques have emerged, making it possible to determine degree of steatosis as well asfibrosis. Despite this,the benefit of routine diagnostics remains uncertain. A better understanding of the（molecular） pathogenesis of NAFLD is needed combined with long-term studies where benefits of treatment can be assessed to determine costbenefit ratios. This review summarizes the current state of knowledge and possible areas of treatment.

Rab proteins implicated in lipid storage and mobilization

Abnormal intracellular accumulation or transport of lipids contributes greatly to the pathogenesis of human diseases. In the liver, excess accumulation of triacylglycerol （TG） leads to fatty liver disease encompassing steatosis, steatohepatitis and fibrosis. This places individuals at risk of developing cirrhosis, hepatocellular carcinoma or hepatic decompensation and also contributes to the emergence of insulin resistance and dyslipidemias affecting many other organs. Excessive accumulation of TG in adipose tissue contributes to insulin resistance as well as to the release of cytokines attracting leucocytes leading to a pro-inflammatory state. Pathological accumulation of cholesteryl ester （CE） in macrophages in the arterial wall is the progenitor of atherosclerotic plaques and heart disease. Overconsumption of dietary fat, cholesterol and carbohydrates explains why these diseases are on the increase yet offers few clues for how to prevent or treat individuals. Dietary regimes have proven futile and barfing surgery, no realistic alternatives are at hand as effective drugs are few and not without side effects. Overweight and obesity-related diseases are no longer restricted to the developed world and as such, constitute a global problem. Development of new drugs and treatment strategies are a priority yet requires as a first step, elucidation of the molecular pathophysiology underlying each associated disease state. The lipid droplet （LD）, an up to now over- looked intracellular organelle, appears at the heart of each pathophysiology linking key regulatory and metabolic processes as well as constituting the site of storage of both TGs and CEs. As the molecular machinery and mechanisms of LDs of each cell type are being elucidated, regulatory proteins used to control various cellular processes are emerging. Of these and the subject of this review, small GTPases belonging to the Rab protein family appear as important molecular switches used in the regulation of the intracellular trafficking and storage of lipids.

Shunts, channels and lipoprotein endosomal traffic： a new model of cholesterol homeostasis in the hepatocyte

The liver directs cholesterol metabolism in the organism. All the major fluxes of cholesterol within the body involve the liver： dietary cholesterol is directed to the liver; cholesterol from peripheral cells goes to the liver; the liver is a major site of cholesterol synthesis for the organism; cholesterol is secreted from the liver within the bile, within apoB lipoproteins and translocated to nascent HDL. The conventional model of cholesterol homeostasis posits that cholesterol from any source enters a common, rapidly exchangeable pool within the cell, which is in equilibrium with a regulatory pool. Increased influx of cholesterol leads rapidly to decreased synthesis of cholesterol. This model was developed based on in vitro studies in the fibroblast and validated only for LDL particles. The challenges the liver must meet in vivo to achieve cholesterol homeostasis are far more complex. Our model posits that the cholesterol derived from three different lipoproteins endosomes has three different fates： LDL-derived cholesterol is largely recycled within VLDL with most of the cholesterol shunted through the hepatocyte without entering the exchangeable pool of cholesterol; high density lipoprotein-derived CE is transcytosed into bile; and chylomicron remnant-derived cholesterol primarily enters the regulatory pool within the hepatocyte. These endosomal channels represent distinct physiological pathways and hepatic homeostasis represents the net result of the outcomes of these distinct channels. Our model takes into account the distinct physiological challenges the hepatocyte must meet, underlie the pathophysiology of many of the apoB dyslipoproteinemias and account for the sustained effectiveness of therapeutic agents such as statins.

Ferroptosis:copper-iron connection in cuprizone-induced demyelination

Redox active metals such as iron,copper,zinc,and manganese play important roles in promoting a variety of biochemical reactions essential for cellular function.This is made possible by the ability of these metals to accept and donate electrons.Iron in the form of iron-sulfur clusters and heme plays a key role in adenosine triphosphate generation in mitochondria as well as numerous other enzymatic reactions.On the other hand.

Drug discovery from traditional Chinese herbal medicine using high content imaging technology

Traditional Chinese medicine(TCM) has been widely used in China and other Asia countries for thousands of years to treat or prevent human diseases. Chinese herbal medicine, one of the most important components of TCM, has unique diversities in chemical components, and thus results in a wide range of biological activities. However, pharmaceutical industry is facing a major challenge to develop a large population of novel natural products and drugs, and considerable efforts have not resulted in highvolume of novel drug discovery and productivity. At present, increasing attention has been paid to Chinese herb medicine modernization in combination with the cutting-age technologies of drug discovery, especially the high throughput selection. High content imaging is an image-based high throughput screening method by using automated microscopy and image analysis software to capture and analyze phenotypes at a large scale to investigate multiple biological features simultaneously in the biological complex. Here, we described the pipeline of the state-of-the-art high content imaging technology, summarized the applications of the high content imaging technology in drug discovery from traditional Chinese herbal medicine, and finally discussed the current challenges and future perspectives for development of high throughput image-based screening technology in novel drug research and discovery.

Elderly patients with inflammatory bowel disease: Updated review of the therapeutic landscape

High-quality data remains scarce in terms of optimal management strategies in the elderly inflammatory bowel disease(IBD)population.Indeed,available trials have been mostly retrospective,of small sample size,likely owing to underrepresentation of such a population in the major randomized controlled trials.However,in the last five years,there has been a steady increase in the number of published trials,helping clarify the estimated benefits and toxicity of the existing IBD armamentarium.In the Everhov trial,prescription strategies were recorded over an average follow-up of 4.2 years.A minority of elderly IBD patients(1%-3%)were treated with biologics within the five years following diagnosis,whilst almost a quarter of these patients were receiving corticosteroid therapy at year five of follow-up,despite its multiple toxicities.The low use of biologic agents in real-life settings likely stems from limited data suggesting lower efficacy and higher toxicity.This minireview will aim to highlight current outcome measurements as it portends the elderly IBD patient,as well as summarize the available therapeutic strategies in view of a growing body of evidence.

Therapeutic drug monitoring in inflammatory bowel disease:The dawn of reactive monitoring

Inflammatory bowel disease(IBD)is a chronic condition that significantly affects the quality of life of its patients.Biologic drugs have been the mainstay treatment in the management of IBD patients but despite their significant contribution,there remains a proportion of patients that do not respond or lose response to treatment.Therapeutic drug monitoring(TDM)involves measuring levels of serum drug concentrations and anti-drug antibodies.TDM of biologic drugs initially emerged to understand treatment failure in other immune mediated inflammatory diseases.This was then introduced in IBD to rationalize primary non-response or secondary loss of response,given that low serum drug concentrations or the formation of anti-drug antibodies are variably associated with treatment failure.The aim of this narrative review is to provide an overview regarding the current use of TDM in clinical practice and to present the evidence available regarding its use in both proactive and reactive clinical settings in preventing and managing treatment failure.This review also presents the existing evidence regarding the association of various clinical outcomes with specific thresholds of drug concentrations,in everyday practice.A narrative review of published articles and conference abstracts regarding the use of TDM in IBD management,through an electronic search using PubMed and ScienceDirect.TDM has proven to be superior and more cost effective in guiding management of patients with treatment failure compared to empiric dose escalation or change in treatment.Despite a trend towards an association between clinical outcomes and drug concentrations,proactive TDM based strategies have not been shown to achieve clear benefit in long-term outcomes.In the clinical setting,TDM has proven to be useful in managing IBD patients,and its use in the reactive setting,as an additional tool to help manage patients with treatment failure,is being promoted as newer guidelines and consensus groups implement TDM as part of the management plan.

Medical research during the COVID-19 pandemic

The current pandemic of coronavirus disease 2019(COVID-19)which was first detected in Wuhan,China in December 2019 is caused by the novel coronavirus named severe acute respiratory syndrome coronavirus-2(SARS-CoV-2).The virus has quickly spread to a large number of countries leading to a great number of deaths.Unfortunately,till today there is no specific treatment or vaccination for SARS-CoV-2.Most of the suggested treatment medications are based on in vitro laboratory investigations,experimental animal models,or previous clinical experience in treating similar viruses such as SARS-CoV-1 or other retroviral infections.The running of any clinical trial during a pandemic is affected at multiple levels.Reasons for this include patient hesitancy or inability to continue investigative treatments due to self-isolation/quarantine,or limited access to public places(including hospitals).Additional barriers relate to health care professionals being committed to other critical tasks or quarantining themselves due to contact with COVID-19 positive patients.The best research approaches are those that adapt to such external unplanned obstacles.Ongoing clinical trials before COVID-19 pandemic have the potential for identifying important therapies in the long-term if they can be completed as planned.However,these clinical trials may require modifications due a pandemic such as this one to ensure the rights,safety,and wellbeing of participants as well as medical staff involved in the conduction of clinical trials.Clinical trials initiated during the pandemic must be time-efficient and flexible due to high contagiousness of severe acute respiratory syndrome coronavirus 2,the significant number of reported deaths,and time constraints needed to perform high quality clinical trials,enrolling adequate sample sizes.Collaboration between different countries as well as implementation of innovative clinical trial designs are essential to successfully complete such initiatives during the current pandemic.Studies looking at the long term sequalae of COVID-19 are also of importance as recent publications describe multi-organ involvement.Long term follow-up of COVID-19 survivors is thus also important to identify possible physical and mental health sequellae.

Eph Receptors and Ephrins in Neuron-Astrocyte Communication at Synapses

神经元-胶质细胞间的相互作用对调节脑内突触联系有重要作用。星形胶质细胞对突触的发育、维持和可塑性有特别关键、复杂的作用。同样,神经元也对星形胶质细胞的生理功能产生影响。但是,神经元和星形胶质细胞之间相互作用的分子机制尚未完全阐明。近来研究表明,Eph受体酪氨酸激酶及轴突导向因子在突触间接触依赖性神经元-胶质细胞的相互作用中起重要作用。与配体结合后,这2个细胞表面相关蛋白家族成员激活双向信号通路,调节神经元和星形胶质细胞的结构和生理特征。本综述着重探讨Eph受体酪氨酸激酶及轴突导向因子在突触间神经元-胶质细胞的相互作用中扮演的角色,并讨论其在突触可塑性、行为及疾病中的潜在作用。