Proteomics of serum exosomes identified fibulin-1 as a novel biomarker for mild cognitive impairment

Mild cognitive impairment(MCI)is a prodrome of Alzheimer’s disease pathology.Cognitive impairment patients often have a delayed diagnosis because there are no early symptoms or conventional diagnostic methods.Exosomes play a vital role in cell-to-cell communications and can act as promising biomarkers in diagnosing diseases.This study was designed to identify serum exosomal candidate proteins that may play roles in diagnosing MCI.Mass spectrometry coupled with tandem mass tag approach-based non-targeted proteomics was used to show the differentially expressed proteins in exosomes between MCI patients and healthy controls,and these differential proteins were validated using immunoblot and enzyme-linked immunosorbent assays.Correlation of cognitive performance with the serum exosomal protein level was determined.Nanoparticle tracking analysis suggested that there was a higher serum exosome concentration and smaller exosome diameter in individuals with MCI compared with healthy controls.We identified 69 exosomal proteins that were differentially expressed between MCI patients and healthy controls using mass spectrometry analysis.Thirty-nine exosomal proteins were upregulated in MCI patients compared with those in control patients.Exosomal fibulin-1,with an area under the curve value of 0.81,may be a biomarker for an MCI diagnosis.The exosomal protein signature from MCI patients reflected the cell adhesion molecule category.In particular,higher exosomal fibulin-1 levels correlated with lower cognitive performance.Thus,this study revealed that exosomal fibulin-1 is a promising biomarker for diagnosing MCI.

Dopamine and cyclic adenosine monophosphate-regulated phosphoprotein with an apparent Mr of 32000 promotes colorectal cancer growth

BACKGROUND Dopamine and cyclic adenosine monophosphate(cAMP)-regulated phosphop-rotein with an apparent Mr of 32000(DARPP-32)is a protein that is involved in regulating dopamine and cAMP signaling pathways in the brain.However,recent studies have shown that DARPP-32 is also expressed in other tissues,including colorectal cancer(CRC),where its function is not well understood.AIM To explore the effect of DARPP-32 on CRC progression.METHODS The expression levels of DARPP-32 were assessed in CRC tissues using both quantitative polymerase chain reaction and immunohistochemistry assays.The proliferative capacity of CRC cell lines was evaluated with Cell Counting Kit-8 and 5-ethynyl-2’-deoxyuridine assays,while apoptosis was measured by flow cytometry.The migratory and invasive potential of CRC cell lines were deter-mined using wound healing and transwell chamber assays.In vivo studies involved monitoring the growth rate of xenograft tumors.Finally,the underlying molecular mechanism of DARPP-32 was investigated through RNA-sequencing and western blot analyses.RESULTS DARPP-32 was frequently upregulated in CRC and associated with abnormal clinicopathological features in CRC.Overexpression of DARPP-32 was shown to promote cancer cell proliferation,migration,and invasion and reduce apoptosis.DARPP-32 knockdown resulted in the opposite functional effects.Mechanistically,DARPP-32 may regulate the phosphoinositide 3-kinase(PI3K)/AKT signaling pathway in order to carry out its biological function.CONCLUSION DARPP-32 promotes CRC progression via the PI3K/AKT signaling pathway.

ZIF-8 coated gold nanospheres: A multi-responsive drug delivery system promoting the killing effect of photothermal therapy against osteosarcoma cells

Photothermal therapy(PTT)has been widely used in the treatment of tumors,but its efficacy is greatly limited by the inability of precise drug delivery and the increase of heat shock proteins(HSPs)caused by high temperature.This article describes a therapeutic strategy to enhance PTT with starvation therapy in conjunction with ferroptosis mechanism.A nanoparticle platform ZIF-8@GA was constructed by wrapping together glucose oxidase(GOX)and gold nanospheres(AuNPs)loaded with dihydroartemisinin(DHA)with zeolitic imidazolate framework-8(ZIF-8).This platform can take advantage of the micro-environment of osteosarcoma(OS)cells,featuring low pH and high reactive oxygen species(ROS),for precision drug delivery.GOX can effectively catalyze glucose to produce gluconic acid and H_(2)O_(2),and DHA can also induce ROS production in OS cells.ROS produced by GOX and DHA can further generate lipid peroxidation(LPO)and lead to ferroptosis of OS cells.At the same time,ROS and LPO produced can inhibit the expression of HSPs,thereby increasing the therapeutic effect of PTT.In vitro experiments show that the nanoparticles are pH and ROS responsive.1μg/mL GOX combined with 0.2μg/mL DHA promotes ferroptosis of OS cells,and increases the killing effect of near-infrared(NIR)on OS cells.Further in vivo experiments showed that the nano drug-delivery platform combined with PTT can effectively inhibit the growth of OS cells.Meanwhile,this study provides a new idea for the treatment of OS with biomaterials combined with various treatment methods.

Exosome-mediated aptamer S58 reduces fibrosis in a rat glaucoma filtration surgery model

AIM:To confirm whether exosome-mediated delivery of aptamer S58(Exo-S58) has a better antifibrotic effect than naked S58 in human conjunctival fibroblasts(HCon Fs) and a rat glaucoma filtration surgery(GFS) model.METHODS:To enhance the effective reaction time of aptamer S58 in vivo, we loaded aptamer S58 into exosomes derived from HEK293 T cells by PEI transfection to determine the effect of Exo-S58 in HCon Fs and a rat GFS model.RESULTS:Exo-S58 can significantly reduce cell proliferation, migration and fibrosis in TGF-β2-induced HCon Fs. In an in vivo experiment, Exo-S58 treatment prolonged filtering bleb retention and reduced fibrosis compared with naked S58 treatment in GFS rats.CONCLUSION:The exosomes are safe and valid carriers to deliver aptamers. Furthermore, Exo-S58 exhibited superior antifibrotic effect than naked S58 both in HCon Fs cells and rat GFS models.

Degradation of helicase-like transcription factor(HLTF)byβ-TrCP promotes hepatocarcinogenesis via activation of the p62/mTOR axis

Helicase-like transcription factor (HLTF) has been found to be involved in the maintenance of genome stability and tumoursuppression, but whether its downregulation in cancers is associated with posttranslational regulation remains unclear. Here, weobserved that HLTF was significantly downregulated in hepatocellular carcinoma (HCC) tissues and positively associated with thesurvival of HCC patients. Mechanistically, the decreased expression of HLTF in HCC was attributed to elevated β-TrCP-mediated ubiquitination and degradation. Knockdown of HLTF enhanced p62 transcriptional activity and mammalian target of rapamycin (mTOR)activation, leading to HCC tumourigenesis. Inhibition of mTOR effectively blocked β-TrCP overexpression- or HLTF knockdownmediated HCC tumourigenesis and metastasis. Furthermore, in clinical tissues, decreased HLTF expression was positively correlatedwith elevated expression of β-TrCP, p62, or p-mTOR in HCC patients. Overall, our data not only uncover new roles of HLTF in HCCcell proliferation and metastasis, but also reveal a novel posttranslational modification of HLTF by β-TrCP, indicating that theβ-TrCP/HLTF/p62/mTOR axis may be a new oncogenic driver involved in HCC development. This finding provides a potentialtherapeutic strategy for HCC patients by targeting the β-TrCP/HLTF/p62/mTOR axis.

Improvement of antibacterial, anti-inflammatory, and osteogenic properties of OGP loaded Co-MOF coating on titanium implants for advanced osseointegration

The bacterial infection,especially for methicillin-resistant Staphylococcus aureus(MRSA),and the associated severe inflammatory response could extremely limit the crosstalk of RAW264.7 cells and mesenchymal stem cells(MSCs)and lead to the undesirable osseointegration of peri–implants.It is highly demanded to modify the surface of titanium(Ti)implant to improve its anti-bacterial and anti-inflammatory properties and facilitate its disabled osseointegration.Herein,in our study,a multifunctional coating of zeolitic imidazolate frameworks-67 encapsulated osteogenic growth peptide(OGP)(ZO)was fabricated on titanium dioxide nanotubes(TNT)substrates(TNT-ZO)via the electrophoresis deposition(EPD)approach.The TNT-ZO substrates exhibited excellent antibacterial activity indicated by the reactive oxygen species(ROS)generation,outer membrane(OM)and inner membrane(IM)permeabilization change,adenosine triphosphate(ATP)decrease,and intracellular compounds(DNA/RNA)leakage.Importantly,the regulation effects of TNT-ZO coating modified titanium substrates on the RAW264.7-MSCs crosstalk could induce the anti-inflammatory and osteogenic microenvironment via multiple paracrine signaling of Runx2,BMP2,VEGF,and TGF-β1.The promoted effects of coating structure were investigated in vivo,including antibacterial effect,osteogenic differentiation of mesenchymal stem cells,and anti-inflammation of RAW264.7 cells,as well as infected bone regeneration and repair in bone defect transplantation model.The results demonstrated that MRSA was effectively eliminated by the hydrolysis of ZIF-67 nanoparticles on TNT-ZO substrates.Furthermore,the excellent osseointegration of peri–implants was realized simultaneously by modulating the crosstalk of RAW264.7-MSCs.This study could provide a novel approach to designing a multifunctional coating on the Ti implants for infected bone regeneration in orthopedic applications.

Mine disaster survivor’s pelvic floor hernia treated with laparoscopic surgery and a perineal approach:A case report

BACKGROUND A pelvic floor hernia is defined as a pelvic floor defect through which the intraabdominal viscera may protrude.It is an infrequent complication following abdominoperineal surgeries.This type of hernia requires surgical repair by conventional or reconstructive techniques.The main treatments could be transabdominal,transperineal or a combination.CASE SUMMARY In this article,we present the case of a recurrent perineal incisional hernia,postresection of the left side of the pelvis,testis and lower limbs resulting from a mine disaster 18 years ago.Combined laparoscopic surgery with a perineal approach was performed.The pelvic floor defect was repaired by a biological mesh and one pedicle skin flap.No signs of recurrence were indicated during the 2 years of follow-up.CONCLUSION The combination of laparoscopic surgery with a perineal approach was effective.The use of the biological mesh and pedicle skin flap to restructure the pelvic floor was effective.

Health management in children with osteogenesis imperfecta

Due to the incurable characteristics of osteogenesis imperfecta,health management plays a crucial role for children in healthy growth,independent life and integrating into society.This paper summarizes three dimensions of "biology-psychology-society",which summarize the research progress for health management in children with osteogenesis imperfecta.In the dimension of biology,the management on diet and complications about children is relatively definite,but more experiments are still needed in order to find out the appropriate values for the using doses of bisphosphonate and treatment time.Additionally,there is a lack of tools to assess the painful degree in children and sports management methods with different types of children with osteogenesis imperfecta nowadays.In the dimension of psychology,it is found that children with osteogenesis imperfecta,their families and carers are all expected to maintain a good state of mind.In the social dimension,we have known the need of children and their families,but their supporting systems are still expected to be improved through practice.

Eligibility of C-BIOPRED severe asthma cohort for type-2 biologic therapies

To the Editor:Patients with severe persistent asthma experience greater morbidity with more impairment in quality of life despite higher use of health care resources and being treated with existing asthma treatments such as inhaled corticosteroids and b-agonists,and sometimes oral corticosteroid(OCS)therapy.Type-2(T2)high asthma has been identified as a phenotype that responds to targeted T2 biologic therapies such as anti-IgE,anti-interleukin(IL)5,or anti-IL5Ra and anti-IL4Ra monoclonal antibodies,which are currently available in Europe and North America,and are currently introduced in the rest of the world.[1]

Effect of D_(4)Dopamine Receptor on Na^(+)-K^(+)-ATPase Activity in Renal Proximal Tubule Cells

Objective:Dopamine,via its receptors,plays a vital role in the maintenance of blood pressure by modulating renal sodium transport.However,the role of the D_(4)dopamine receptor(D_(4)receptor)in renal proximal tubules(PRTs)is still unclear.This study aimed to verify the hypothesis that activation of D_(4)receptor directly inhibits the activity of the Na+-K+-ATPase(NKA)in RPT cells.Methods:NKA activity,nitric oxide(NO)and cyclic guanosine monophosphate(cGMP)levels were measured in RPT cells treated with the D_(4)receptor agonist PD168077 and/or the D_(4)receptor antagonist L745870,the NO synthase inhibitor NG-nitro-L-arginine-methyl ester(L-NAME)or the soluble guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one(ODQ).Total D_(4)receptor expression and its expression in the plasma membrane were investigated by immunoblotting in RPT cells from Wistar-Kyoto(WKY)rats and spontaneously hypertensive rats(SHRs).Results:Activation of D_(4)receptors with PD168077,inhibited NKA activity in RPT cells from WKY rats in a concentration-and time-dependent manner.The inhibitory effect of PD168077 on NKA activity was prevented by the addition of the D_(4)receptor antagonist L745870,which by itself had no effect.The NO synthase inhibitor L-NAME and the soluble guanylyl cyclase inhibitor ODQ,which by themselves had no effect on NKA activity,eliminated the inhibitory effect of PD168077 on NKA activity.Activation of D_(4)receptors also increased NO levels in the culture medium and cGMP levels in RPT cells.However,the inhibitory effect of D_(4)receptors on NKA activity was absent in RPT cells from SHRs,which could be related to decreased plasma membrane expression of D_(4)receptors in SHR RPT cells.Conclusions:Activation of D_(4)receptors directly inhibits NKA activity via the NO/cGMP signaling pathway in RPT cells from WKY rats but not SHRs.Aberrant regulation of NKA activity in RPT cells may be involved in the pathogenesis of hypertension.

Knockdown of hepatocyte Perilipin-3 mitigates hepatic steatosis and steatohepatitis caused by hepatocyte CGI-58 deletion in mice

Comparative gene identification-58(CGI-58),also known asα/βhydrolase domain containing 5,is the co-activator of adipose triglyceride lipase that hydrolyzes triglycerides stored in the cytosolic lipid droplets.Mutations in CGI-58 gene cause Chanarin-Dorfman syndrome(CDS),an autosomal recessive neutral lipid storage disease with ichthyosis.The liver pathology of CDS manifests as steatosis and steatohepatitis,which currently has no effective treatments.Perilipin-3(Plin3)is a member of the Perilipin-ADRP-TIP47 protein family that is essential for lipid droplet biogenesis.The objective of this study was to test a hypothesis that deletion of a major lipid droplet protein alleviates fatty liver pathogenesis caused by CGI-58 deficiency in hepatocytes.Adult CGI-58-floxed mice were injected with adeno-associated vectors simultaneously expressing the Cre recombinase and microRNA against Plin3 under the control of a hepatocyte-specific promoter,followed by high-fat diet feeding for 6 weeks.Liver and blood samples were then collected from these animals for histological and biochemical analysis.Plin3 knockdown in hepatocytes prevented steatosis,steatohepatitis,and necroptosis caused by hepatocyte CGI-58 deficiency.Our work is the first to show that inhibiting Plin3 in hepatocytes is sufficient to mitigate hepatocyte CGI-58 deficiency-induced hepatic steatosis and steatohepatitis in mice.

Expert consensus on organizing the multidisciplinary team(MDT) diagnosis and treatment of hepato-pancreato-biliary diseases in China

Multidisciplinary team(MDT) discussion has been well established in some European countries and widely carried out in various clinical professional fields(Festen et al.,2021;Fleissig et al.,2006).Many studies have reported the benefits of MDT,including improving survival and quality of cancer care(Munro et al.,2015;Nixon et al.,2021;Tsai et al.,2020).

Guidelines for the Diagnosis and Treatment of Rosacea in China(2021 Edition)

Rosacea is a chronic inflammatory skin disease that primarily affects the centrofacial areas and mainly manifests as recurrent flushing and erythema.In recent years,there has been progress in the understanding of the diagnosis and treatment of rosacea.Therefore,a group of dermatological experts updated the guidelines based on the 2016 expert consensus statement on rosacea diagnosis and treatment in China.These new guidelines propose diagnostic criteria for rosacea at different sites to further standardize the diagnosis and treatment of rosacea in China.

Chinese Guidelines for the Diagnosis and Treatment of Urticaria:2018 Update

Urticaria is a common dermatological condition that is characterized by wheals and pruritus.Certain types of urticaria can be resistant to treatment,and recurrence is common.These guidelines supplement and improve upon the previous version(published in 2014)of the Guidelines for the Diagnosis and Treatment of Urticaria in China,and incorporate recent research advances in urticaria.These guidelines are suitable for both Chinese adults and children of Han nationality with urticaria.These guidelines update and broaden the definition,pathogenesis,classification,diagnosis,and treatment of urticaria,and serve as a scientific and authoritative reference for the diagnosis and management of urticaria.

Comparison of survival prediction values of different scoring models for patients undergoing transjugular intrahepatic portal shunt:A multicenter retrospective study

Aim:The aim of this study is to compare the prognostic values of the Child–Pugh,integrated model for end‐stage liver disease(iMELD),albumin–bilirubin(ALBI),and Freiburg index of postsurvival(FIPS)scores in patients undergoing transjugular intrahepatic portosystemic shunt(TIPS).Methods:We conducted a multicenter retrospective study including patients who underwent TIPS by collecting data from several hospitals in southwest China between January 2014 and February 2021.We compared the performance of different scoring models for survival prediction in these patients.The performance of each scoring model was assessed via area under the receiver‐operating characteristic(AUROC)curve analysis.Results:The study included 378 TIPS patients(268 men,110 women;median age 52[interquartile range,45–60]years).Age;cirrhosis etiology;ascites severity;albumin levels;international normalized ratio;total bilirubin levels;sodium levels;and Child–Pugh,iMELD,ALBI,and FIPS scores were significant prognostic factors in cirrhotic patients who underwent TIPS.The Child–Pugh,iMELD,ALBI,and FIPS scores were all independent predictors of survival in TIPS patients.Survival analysis showed that all scoring models effectively stratified the prognostic risk of these patients.The Child–Pugh score was the best predictor of postoperative survival,followed by the ALBI and FIPS scores.The iMELD score was the worst predictor.The Child–Pugh,iMELD,ALBI,and FIPS scores predicted the 1‐year postoperative survival,with AUROC values of 0.832,0.677,0.761,and 0.745,respectively,and the 3‐year postoperative survival,with AUROC values of 0.710,0.668,0.721,and 0.658,respectively.The calibration curve showed that the Child–Pugh,ALBI,and FIPS models performed well in predicting 1‐and 3‐year survival,whereas the iMELD model was a poor predictor.Conclusions:The four scoring models can predict survival in cirrhotic patients after TIPS and can effectively stratify prognostic risk.The Child–Pugh score may be more suitable for predicting survival after TIPS in patients with liver cirrhosis.

Chinese Guidelines for the Management of Chronic Pruritus(2018)

Chronic pruritus is a very common clinical symptom that seriously affects people’s quality of life.Pruritus is associated not only with skin diseases but also with a variety of systemic diseases.It brings great challenges to clinical management.To standardize the management of chronic pruritus,experts of the Allergic Disease Committee of Chinese Dermatologist Association discussed many times and finally formed this guideline to provide reference and guidance for the clinical work of dermatologists and other physicians at all levels.

Chinese Expert Consensus on Clinical Application of Patch Test (Revised 2020)

Allergic contact dermatitis is a common clinical allergic disease.The patch test is the gold standard for finding and clarifying contact allergens.With the deepening understanding of the epidemiology,pathogenesis,and clinical manifestations of contact dermatitis,as well as the increased requirements for the standardized application of patch tests,the relevant consensus needs to be continuously updated.This consensus is updated on the basis of the version issued in 2015.In this revised version,it refines the selection of clinical indications,improves the judgment end interpretation of results,adds delayed detection and patient education,and so on,which aims to further standardize clinical applications and improve the value of patch test.

The near-infrared dye IR-61 restores erectile function in a streptozotocin-induced diabetes model via mitochondrial protection

This study aimed to evaluate the therapeutic effect of IR-61,a novel mitochondrial heptamethine cyanine dye with antioxidant effects,on diabetes mellitus-induced erectile dysfunction(DMED).Eight-week-old male Sprague-Dawley rats were intraperitoneally injected with streptozotocin(STZ)to induce type 1 diabetes.Eight weeks after STZ injection,all rats were divided into three groups:the control group,DM group,and DM+IR-61 group.In the DM+IR-61 group,the rats were administered IR-61(1.6 mg kg-1)twice a week by intravenous injection.At week 13,erectile function was evaluated by determining the ratio of the maximal intracavernous pressure to mean arterial pressure,and the penises were then harvested for fluorescent imaging,transmission electron microscopy,histological examinations,and Western blot analysis.Whole-body imaging suggested that IR-61 was highly accumulated in the penis after intravenous injection.IR-61 treatment significantly improved the maximal ICP of diabetic rats.Additionally,IR-61 ameliorated diabetes-induced inflammation,apoptosis,and phenotypic transition of corpus cavernosum smooth muscle cells(CCSMCs)in penile tissue.IR-61 also attenuated mitochondrial damage,reduced reactive oxygen species production in the corpus cavernosum and upregulated sirtuinl(SIRT1),sirtuin3(SIRT3),nuclear factor(erythroid-derived 2)-like 2(Nrf2),and heme oxygenase expression in penile tissue.In conclusion,IR-61 represents a potential therapeutic option for DMED by protecting the mitochondria of CCSMCs,which may be mediated by activation of the SIRT1,SIRT3,and Nrf2 pathways.

The E3 ubiquitin ligase HUWE1 acts through the N-Myc-DLL1-NOTCH1 signaling axis to suppress glioblastoma progression

Background:Elucidation of the post-transcriptional modification has led to novel strategies to treat intractable tumors,especially glioblastoma(GBM).The ubiquitin-proteasome system(UPS)mediates a reversible,stringent and stepwise post-translational modification which is closely associated with malignant processes of GBM.To this end,developing novel therapeutic approaches to target the UPS may contribute to the treatment of this disease.This study aimed to screen the vital and aberrantly regulated component of the UPS in GBM.Based on the molecular identification,functional characterization,and mechanism investigation,we sought to elaborate a novel therapeutic strategy to target this vital factor to combat GBM.Methods:We combined glioma datasets and human patient samples to screen and identify aberrantly regulated E3 ubiquitin ligase.Multidimensional database analysis and molecular and functional experiments in vivo and in vitro were used to evaluate the roles of HECT,UBA and WWE domain-containing E3 ubiquitin ligase 1(HUWE1)in GBM.dCas9 synergistic activation mediator system and recombinant adeno-associated virus(rAAV)were used to endogenously overexpress full-length HUWE1 in vitro and in glioma orthotopic xenografts.Results:Low expression of HUWE1 was closely associated with worse prognosis of GBM patients.The ubiquitination and subsequent degradation of N-Myc mediated by HUWE1,leading to the inactivation of downstream Delta-like 1(DLL1)-NOTCH1 signaling pathways,inhibited the proliferation,invasion,and migration of GBM cells in vitro and in vivo.A rAAV dual-vector system for packaging and delivery of dCas9-VP64 was used to augment endogenous HUWE1 expression in vivo and showed an antitumor activity in glioma orthotopic xenografts.Conclusions:The E3 ubiquitin ligase HUWE1 acts through the N-Myc-DLL1-NOTCH1 signaling axis to suppress GBM progression.Antitumor activity of rAAV dual-vector delivering dCas9-HUWE1 system uncovers a promising therapeutic strategy for GBM.

Chinese Experts Consensus on Biologic Therapy for Psoriasis

Biologic drugs have been increasingly used in the treatment of psoriasis and are especially favorable for severe,recalcitrant,and special-type cases.Therefore,appropriate,effective,and safe use of biologic drugs has drawn attention from dermatologists.For this purpose,Chinese experts majoring in psoriasis analyzed domestic and international research data,summarized current clinical experiences,investigated features of Chinese patients with psoriasis,and finally formulated the present consensus,which provides detailed guidances to clinicians in terms of the principles and methods of the application of biologics,the efficacy and safety profile,patient screening and monitoring,main adverse events and corresponding solutions,and precautions for special patient populations.